5 (2008)


REVIEW The Biochemistry of Drug Metabolism – An Introduction
Part 4. Reactions of Conjugation and Their Enzymes
by Bernard Testa* a ) and Stefanie D. Krämer b )

) Department of Pharmacy, University Hospital Centre (CHUV), Rue du Bugnon, CH-1011 Lausanne (e-mail: b ) Department of Chemistry and Applied Biosciences, ETH Zurich, Wolfgang-Pauli-Strasse 10, CH-8093 Zurich

This Part 4 of our biochemical introduction to drug metabolism [1 – 4] presents the reactions of conjugation and their enzymes. As we shall see, reactions of conjugation are also a major focus of interest in the metabolism of drugs and other xenobiotics. Books specifically dedicated to conjugation reactions are rare [5], but much recent information can be found in book chapters (e.g., [6] [7]). For a reaction of conjugation to occur, a suitable functional group must be present in the substrate, which will serve as the anchoring site for an endogenous molecule or moiety such as CH3 , sulfate, glucuronic acid, or glutathione. Conjugation reactions are thus synthetic (i.e., anabolic) reactions whose products are of modestly to markedly higher molecular weight than the corresponding substrate. As for the anchoring group, it can either be present in a xenobiotic or be created by a functionalization reaction. In other words, reactions of conjugation are able to produce first-generation as well as later-generation metabolites. We, therefore, consider as unfelicitous the term of phase II reactions commonly used to designate conjugations. A first issue when discussing reactions of conjugation will be to offer a clear definition. As we shall see, a number of criteria exist, all of which show some degree of fuzziness, and only one of which must necessarily be met. This has indeed led to some confusion with reactions of hydrolysis, which some biochemists have viewed as conjugation. We oppose such a view for reasons previously explained [3]. To repeat what we stated, reactions of hydrolysis are not catalyzed by transferases (EC 2) but by hydrolases (EC 3) [8], and water is not an endogenously synthesized molecule or moiety linked covalently to a cofactor. Reactions of conjugation, like the reactions of functionalization we saw in Parts 2 and 3, act on exogenous substrates (i.e., xenobiotics [1]) as well as endogenous substrates (i.e., endobiotics). This dual functionality may create a potential for metabolic interaction between a drug and an endogenous substrate, a frequently overlooked mechanism of toxicity. Thus, there may be competitive affinity for the catalytic site of an endobiotic-metabolizing enzyme, or there may be competition for the limited supply of a cofactor. A typical example of the latter case is found with 
2008 Verlag Helvetica Chimica Acta AG, Zürich



paracetamol, a high-dose drug undergoing extensive glucuronidation whose administration is forbidden to neonates and babies, since it deprives them of the glucuronic acid they need to detoxify bilirubin.



Fig. 4.1. The structure of this Part follows custom as much as logic. First, an introductory Chapter will present an overview of the reactions of conjugation, their criteria, and their similarities and differences with functionalization reactions. As for the major reactions to be discussed in the subsequent Chapters, there is no overwhelming argument for preferring one order over another. We shall begin with the rather straightforward case of the reactions of methylation (Chapt. 4.2). Reactions of sulfonation (Chapt. 4.3) and glucuronidation (Chapt. 4.4) sometimes compete for the same substrates and will, therefore, be treated in sequence. Together with sulfonation, we will have a few things to say about reactions of phosphorylation, whose rarity should not obscure their significance in the activation of some drugs. Chapt. 4.5 and 4.6 center on coenzyme A, but with a difference. Reactions of acetylation (Chapt. 4.5) follow the usual pattern in having the conjugating moiety carried by the coenzyme, here coenzyme A. In contrast, there is a variety of reactions where the substrate (be it a xenobiotic or an endobiotic) is coupled to coenzyme A prior to being processed along vastly different pathways (Chapt. 4.6). Chapt. 4.7 presents glutathione and its reactions, a topic of marked biocomplexity and great toxicological significance. A few unclassifiable reactions will be summarized in Chapt. 4.8.

2174 CHEMISTRY & BIODIVERSITY – Vol. they will be discussed here. 4. they represent poorly investigated pathways nevertheless worthy of some attention. 1. Reference is made to Chapt. This Figure is a slightly amended form of Fig. with a few additional details.8 to help the readers get a better view of the present Part.2 – 4. Two of these pathways are not conjugations stricto sensu and. 4. Nevertheless. As for the reactions in Chapt.8. .12 we saw in Part 1 [1]. 5 (2008) Fig. The focus here is on the conjugating moiety being transferred to the substrate as a result of a conjugation reaction. are written in italics. We also have here a first glance at the different pathways which xenobiotic – coenzyme A conjugates can follow. 4.2. since a coenzyme A conjugate is the indispensable intermediate. for this reason. These two pathways are the unidirectional inversion of configuration of profens (and a few other xenobiotics) and the b-oxidation of fatty acid analogs.

And they are not necessary. typical reactions of oxidoreduction). the endogenous conjugating moiety is usually carried by a cofactor. there may be arbitrariness in deciding whether a conjugating moiety such as CO2 is endogenous.CHEMISTRY & BIODIVERSITY – Vol. Second. the size of the endocon is generally in the range of 100 – 300 Da. It is important from a biochemical and practical viewpoint to note that Criteria 2 – 5 considered separately are neither sufficient nor necessary to define conjugation reactions. Third. This is the absolute criterion of conjugation reactions. Fifth. although.. but there are exceptions. with the chemical bond linking the cofactor and the endocon being a high-energy one such that the Gibbs energy released upon its cleavage drives the transfer of the endocon to the substrate. since. They are not sufficient. First and above all. conjugation reactions are catalyzed by enzymes known as transferases (EC 2) which bind the substrate and the cofactor in such a manner that their close proximity allows the reaction to proceed. Fourth. the hydride is also transferred from a cofactor (NADPH or NADH).3. The metaphor of transferases being a nuptial bed has not escaped some biochemists. 5 (2008) 2175 Fig. this endogenous molecule or moiety is generally polar (hydrophilic) or even highly polar. they involve an endogenous molecule (called the endogenous conjugating moiety. in hydrogenation reactions (i. and sometimes abbreviated as the endocon) with which the substrate is coupled. 4.e. since they all suffer from some important exceptions (see next Figure). Conjugation reactions are characterized by a number of criteria which are presented here [5] [6] [9]. . as we shall see.

is also an exception to Criterion 3 as is the acetyl moiety. since all conjugating moieties involved are indeed endogenous. 4. 4. carbonyl compounds (Chapt. 4. exceptions are glutathione (Chapt.7). 4. 4. the C2 unit in chain elongation is derived from acetyl-coenzyme A (Chapt. 4. catalysis by a transferase (Criterion 5) is almost always the case.6.2176 CHEMISTRY & BIODIVERSITY – Vol. The transfer of a CH3 group is special. . Criterion 1 is indeed the only one that knows no exception. But we also have relatively large endocons such as sterols (Chapt. and the C2 chain elongation (Chapt. 4. 4. Thus. being small. namely the coupling of xenobiotic carboxylic acids to sterols or to diglycerides (to yield mixed triglycerides). 5 (2008) Fig. 4. As can be seen. the few exceptions being the coupling of hydrazines with carbonyl compounds (Chapt. and all reactions in Chapt.8) is clearly also produced in vivo. The criterion of polarity of the endocon (Criterion 2) knows only two major exceptions. since here and as stated it is the substrate rather than the endocon that is attached to coenzyme A. The CH3 group.4. As for the conjugating moiety being carried by a coenzyme (Criterion 4). 4. Finally.6). 4. Most of the CO2 used in the formation of carbamic acids (Chapt.7).6).8) and some nonenzymatic conjugations with glutathione (Chapt.6) and glutathione (Chapt. since it adds a hydrophobic moiety except when forming quaternary ammonium metabolites (Chapt. 4.2).7). This Figure summarizes in tabular form the cases of compliance and noncompliance to the five conjugation criteria.8).

or catabolism by b-oxidation. The latter catalyzes the transfer of the adequate conjugating moiety from the cofactor to the substrate.. The case of noradrenaline is different. an endogenous compound (e.5. or a nutrient (e. a toxic breakdown product of hemoglobin). a fatty acid) is captured by a transferase. 5 (2008) 2177 Fig.CHEMISTRY & BIODIVERSITY – Vol. whose coenzyme A conjugates can undergo anabolism by C2 chain elongation. Their graphical similarity with Fig. bilirubin. the neurotransmitter noradrenaline).23 in Part 1 [1] is not fortuitous. As shown here. the resulting glucuronide being excreted in the bile..g. a waste product of physiological metabolism (e. The case of metabolic intermediates in anabolism (synthetic metabolism) and catabolism (breakdown metabolism) is illustrated with fatty acids.g. 1.. This Figure and the next one illustrate the deep analogy between the physiological reactions of endobiotic conjugation and the conjugation of xenobiotics. . Thus. being N-methylated to the neurotransmitter adrenaline or O-methylated to an inactive metabolite. yielding a conjugate.g. 4. as classified in the Figure. These reactions have evolved to fulfill a variety of functions. the toxic bilirubin is detoxified by conjugation with glucuronic acid.

Similarly. toxicity is usually greatly decreased by conjugation (e. but.6. 4. while others have diversified and are specialized to some extent toward endobiotics or xenobiotics (e..g. glucuronides and glutathione derivatives) is coupled to their active excretion [11]. 5 (2008) Fig.2178 CHEMISTRY & BIODIVERSITY – Vol. there are numerous examples of conjugations leading to toxification. As a rule.. drug conjugation inactivates the substrate. some mixed triglycerides).. N-methylpyridinium). as we shall see.. some transferases recognize endobiotics and xenobiotics alike (e.g. but there are only few noteworthy exceptions such as the highly active morphine 6-O-glucuronide. Some conjugates may indeed be reactive (e. is captured and metabolized by a transferase. whereas others are highly lipophilic and may accumulate in tissues as residues (e. or a phase I metabolite.g. catechol O-methyltransferase). Thus. Reactions of xenobiotic conjugation have evolved from physiological conjugations to fulfill protective functions [10]..g. a xenobiotic containing an adequate target group. UDP-glucuronyltransferases).. such that the formation of polar conjugates (e.g.g. a co-evolution of transferases and transporters is believed to have occurred. Such exceptions should not hide the fact that the greatly increased hydrophilicity of many conjugates relative to their parent compound facilitates their excretion. As we shall see. some acyl glucuronides). What is more. .



Fig. 4.7. This Figure opens Chapt. 4.2 dedicated to biomethylation. These reactions are catalyzed by methyltransferases (EC 2.1.1), and the endogenous conjugating moiety is a CH3 group carried by the cofactor S-adenosyl-l-methionine (4.1; SAM, AdoMet) [5 – 8] [12] [13]. The CH3 group in SAM is bound to a sulfonium center, giving it a marked electrophilic character and explaining its reactivity. During the reaction, S-adenosyl-lmethionine loses the CH3 group and the positive charge to become S-adenosyl-lhomocysteine (4.2). As shown in the Figure, three major types of reactions are recognized, namely the O-methylation of phenolic groups (mainly catechols), the Nmethylation of endocyclic or exocyclic amino groups, and the S-methylation of thiols. Arsenic methylation (not shown in this Figure) will also be discussed. An important aspect made explicit here is the fate of the positive charge carried by the sulfonium center. In most cases, the positive charge is lost in the form of a proton and the Nmethylated metabolite is more lipophilic than the parent compound. The exception is the N-methylation of pyridine-type N-atoms, which forms a quaternary ammonium group retaining the positive charge. This is pharmacokinetically relevant, since such positively charged metabolites are markedly more hydrophilic than the parent compound, resulting in accelerated excretion.



Fig. 4.8. The main enzymes responsible for O-methylation are catechol O-methyltransferase (COMT) and phenol O-methyltransferase (PMT) [8]. The former is far more significant as far as xenobiotic metabolism is concerned, and little can be found about the latter in the relevant literature. COMT is mainly cytosolic (molecular weight of ca. 25 kDa) but also exists in membrane-bound form [13 – 16]. Both forms are products of a single gene, the membrane-bound form including an additional 50 residue segment at the N-terminus. The cytosolic form is expressed to high levels in the liver and kidneys, and the membrane-bound form predominates in the brain. The expression of COMT in human red blood cells has greatly facilitated its study. COMT fulfils important physiological functions by methylating (and inactivating) the catecholamine neurotransmitters dopamine, noradrenaline (norepinephrine), and adrenaline (epinephrine). As we shall see, it also methylates catecholestrogens. Its genetically reduced activity in ca. 25% of Caucasians may have therapeutic and physiopathological significance [13] [17].



Fig. 4.9. O-Methylations are common reactions of compounds containing a catechol moiety, 4.3, with a usual regioselectivity for the meta-position (i.e., 4.4) over the paraposition (i.e., 4.5) [18]. The substrates can be xenobiotics and particularly drugs, as exemplified in the lower part of the Figure which summarizes a few results from an extensive investigation in which the rates of O-methylation of ca. 50 substrates were determined in the presence of recombinant human soluble COMT [18a]. In this Figure, selected substrates are arranged according to their rate of methylation by human recombinant soluble COMT. Very good substrates were for example 4-nitrocatechol (4.6) and caffeic acid (4.7). Fair substrates were catechol itself (4.8) and the drug dobutamine (4.9), a cardiostimulant b1-adrenoceptor agonist. The last row shows three well-known substrates whose rate of methylation was comparatively slow in vitro, namely the neurotransmitter dopamine (4.10), the anti-Parkinsonian drug l-DOPA (4.11), and (S)-carbidopa (4.12), a peripheral inhibitor of l-DOPA decarboxylase used in combination therapy to increase the efficacy of l-DOPA. Structure – metabolism relationship studies showed that bulky substituents in adjacent positions decreased methylation, whereas increased acidity of a OH group favored it. A more refined analysis showed that increased ionization enhanced affinity for COMT, while the turnover rate and Vmax were favored by a high electron density on the phenolate Oatom, in other words, by a restricted delocalization of the negative charge. This is consistent with nucleophilicity of the target group favoring its methylation, in agreement with the electrophilic character of the CH3 group in S-adenosyl-lmethionine [18].

Interestingly. 4.13) is a selective N-methyl-S-aspartate receptor antagonist of potential interest in neurodegenerative diseases and brain injury. Another informative example is that of duloxetine (4.10. . Incubations with human or rodent soluble COMT revealed the fast methylation of the 4’’-OH group. each of which is a potential target for COMT. This inhibitor of serotonin and noradrenaline reuptake contains a naphthalen-1-yl moiety which undergoes oxidation in humans at the 4-. 5. This compound shows two trihydroxyphenyl moieties. A significant metabolite so produced is the catechol derivative 5. One of the major metabolic pathways of the drug in humans is a CYP2D6-catalyzed oxidation to the catechol metabolite. Thus.16). which was found to undergo O-methylation at either position with a predominance for the 6-OH group.17).15). after they have been oxidized to catechols.or 6-positions. 5 (2008) Fig. followed by meta-O-methylation to 3’-methoxytraxoprodil (4. This reaction was followed in a second step by the formation of the 4’. traxoprodil (4. these two metabolites were found to be strong noncompetitive inhibitors of COMT. In addition to xenobiotic substrates. possibly potentiating the activity of endogenous catecholamines [22].17 detected in the urine of humans and rodents.14) [19].6-dihydroxyduloxetine (4. namely the tea polyphenol (À)-epigallocatechin gallate (4. O-methylation can occur as a late event in the metabolism of phenolic or aryl groups.2182 CHEMISTRY & BIODIVERSITY – Vol.4’’-dimethylated metabolite [21]. the major metabolite of 4. The last example in this Figure is taken from phytochemistry. followed by further oxidation [20]. This is exemplified here with two recently introduced drugs.

20.21 and 4. .19 is so fast that this catecholestrogen is in fact a prodrug of 2methoxyestradiol (4. 5 (2008) 2183 Fig.22.11.and 4-hydroxyestradiol (4. In this context.). 4. it is important to note that COMT-catalyzed O-methylation is a protective pathway which decreases quinone formation by competing with oxidoreductases for the catechol substrates. 17b-estradiol (4. These are reactive endogenous metabolites known to react with nucleic acids and to play a role in estrogen carcinogenesis [23]. Catechol O-methyltransferase is also of interest in the detoxification of catechol estrogens.23) attenuates cardiovascular and renal diseases. Of further interest in catecholestrogen O-methylation is the fact that 2-methoxyestradiol (4. Indeed.CHEMISTRY & BIODIVERSITY – Vol. the 17-keto analogue of 4.18) is oxidized by cytochrome P450 to 2hydroxy. These effects were first detected after administration of 2-hydroxyestradiol (4. resp. More recent in vivo studies in rats have revealed that the 2-O-methylation of 4. The catecholestrogens are oxidized by CYPs or peroxidases (PER) to the catecholestrogen quinones 4.19 and 4.18.19).23) [24]. The same pathways are known for estrone.

4.. However. Several enzymes catalyze reactions of xenobiotic Nmethylation with different substrate specificities. nicotinamide NMT is a useful detoxification enzyme. Compared to HNMT.g.15).. 5 (2008) Fig.g. nicotinamide N-methyltransferase (NNMT). N-Methylation is a common pathway for several neurotransmitters and hormones. but its marked interindividual variability also implies that detoxification of pyridine-type compounds is reduced in a fraction of the human population. In other words. (À)-(S)-nicotine (see Fig. a number of xenobiotic amines are known to inhibit it. 4. Besides these endogenous compounds. a significant number of exogenous amines are N-methylated. phenylethanolamine N-methyltransferase. e. . 4. As we shall see. both of which are cytosolic and are expressed in the liver and in a number of other organs.12. This is a worrying cause of side-effects. all the more so since the enzyme is polymorphic in humans. nicotinamide NMT is rather promiscuous and N-methylates a remarkable variety of xenobiotic aromatic azaheterocycles (see Fig.2184 CHEMISTRY & BIODIVERSITY – Vol. Histamine NMT has a narrow substrate specificity and plays a limited role in drug metabolism. the products are quaternary ammonium cations whose polarity and renal clearance are increased compared to the parent compound. e. and nonspecific amine N-methyltransferase [8] [12] [13]. histamine N-methyltransferase (HNMT).15). This Figure presents an Enzyme Identity Card of NNMT and HNMT.

Its role in xenobiotic metabolism is further increased by its wide distribution in organs and tissues. namely phenylethanolamine N-methyltransferase (PNMT) and the nonspecific amine Nmethyltransferase (NMT) also known as indolethylamine N-methyltransferase (INMT) [8]. amine NMT exists as two or more isozymes with broad and overlapping substrate specificities which include primary and secondary aromatic amines as well as aromatic azaheterocycles [25] [26]. Phenylethanolamine NMT is a highly specialized enzyme which at best plays a very limited role in xenobiotic metabolism due to its restricted location (mainly the adrenal medulla) and narrow substrate specificity (noradrenaline and analogous phenylethanolamines) [13]. 4. However. . much remains to be understood about this enzyme. 5 (2008) 2185 Fig. In contrast. especially in the lung.CHEMISTRY & BIODIVERSITY – Vol.13. This Enzyme ID card features two other N-methyltransferases. and particularly the structural variety of substrates and inhibitors it recognizes.

33) [29]. This metabolite is a precursor whose oxidation by monoamine oxidase yields the dopaminergic neurotoxic cation Nmethylisoquinolinium (4. since N-oxygenation here yields a nontoxic N-oxide. Carcinogenic aromatic primary amines such as 4-amino-1. This reaction goes countersense to the well-known oxidative N-demethylation reactions of methylxanthines [2].28) and benzidine (4. 2. In contrast.g. Another toxicologically significant reaction of N-methylation is that of theophylline (4.4-tetrahydroisoquinoline (4.25) itself.2. Tetrahydroisoquinolines have been the objects of numerous investigations due to their potential toxification ultimately yielding neurotoxic ammonium cations showing a close analogy to methyl-4-phenylpyridinium (MPP þ .3. since the secondary amine (e.27).30 [25] is a detoxification.29 to the tertiary amine 4. Various endogenous and exogenous close analogues have been shown to undergo the same sequence of toxification [28b].14. This is well-illustrated by 1. see Fig.31) are also substrates of amine NMT. it is effective in neonates (5 – 10% of a dose of theophylline) and may cause unwanted side-effects. .29) may then be oxidized by flavin monooxygenase to a hydroxylamine [2].94 in Part 2 [2]) [27].2186 CHEMISTRY & BIODIVERSITY – Vol. 4. 4.. In contrast. Their first N-methylation is considered a reaction of toxification. an endogenous and exogenous cyclic secondary amine whose N-methylation (seemingly by amine NMT) yields the tertiary amine 4.32) to yield caffeine (4. 5 (2008) Fig.26 [28a].1’-biphenyl (4. the documented N-methylation of 4. and it is not seen in adult humans.

34). The N-methylation of pyridine is clearly a reaction of detoxification. A particularly interesting example is provided by nicotine [31] [32]. and indeed there exists a trend across animal species such that the more extensive its N-methylation. 5 (2008) 2187 Fig.41) was a potent competitive inhibitor of (R)-nicotine N-methylation. This enzyme is the only one to utilize nicotinamide as a CH3 acceptor.38) are also good substrates. phthalazine (4.15. and quinoxaline (4. Whereas the unnatural (þ)-(R)-nicotine enantiomer (4.35. 4. can be N-methylated to yield a quaternary ammonium cation. amine NMT appears as the major but not only enzyme acting on pyridine (4. In this Figure. the lower its toxicity. 4. whereas here (S)-nicotine was a weak substrate.CHEMISTRY & BIODIVERSITY – Vol.35 (R = H) [26] [30]. Our first example is quite naturally nicotinamide (4. the physiological substrate of nicotinamide N-methyltransferase [12a].39) has been found to be a good substrate of amine N-methyltransferase from guinea pig lung to yield the quaternary ammonium compound 4. . as mentioned above. the natural (À)-(S)nicotine (4.37). Indeed. A number of other azaheterocyclic compounds including quinoline (4. The results in the cytosol of human liver cells again showed (R)-nicotine to be a good substrate. but it also N-methylates other pyridine compounds.40. we exemplify the case of aromatic azaheterocycles. which.36). R ¼ H) and pyridine analogues.

and that markedly more medicinal substrates of the former are known. S-Methyltransferases conjugate thiols into thiomethyl sulfides (Fig.96.1. This enzyme methylated thioethers (i. no gene for TMT has been identified in humans. . despite these differences and the fact that both enzymes are polymorphic in humans. In other words. TEMT) which is briefly mentioned here. 5 (2008) Fig.1. Besides these two enzymes.. the two major enzymes in humans and animals being thiol methyltransferase (TMT) and thiopurine methyltransferase (TPMT). 4. Few if any medicinal examples have been reported. RÀS þ (CH3 )ÀR’). Yet. In particular and at the time of this writing. and it also acts on ether-type selenium compounds.2188 CHEMISTRY & BIODIVERSITY – Vol.. the clinical significance of thiol MT appears greater than that of thiopurine MT. studies on the genetics and pharmacogenetics of TPMT still exceed those on TMT [13] [33].e. Whereas no direct comparison between the two enzymes seems to have been published.16. there exists also a thioether methyltransferase (EC 2.7). 4.e. it is clear that thiol MT shows a much wider substrate specificity than thiopurine MT. RÀSÀR’) to form sulfonium cations (i.

5 (2008) 2189 Fig. the first marketed inhibitor of angiotensin-converting enzyme (ACE). A high activity has been characterized in human hepatic and renal microsomes. plus a number of later-generation Smethylated metabolites such as compound 4. In humans and laboratory animals. 4. since a comparable metabolic pattern was obtained with gemopatrilat. omapatrilat (4.45. most of these compounds are substrates of thiol MT. . 4.16. 4.44). we survey a number of xenobiotic thiols whose Smethylation has been investigated. The second example is a more recent one. A first and well-known example is that of captopril (4. this compound undergoes S-methylation to yield the first-generation metabolite 4. As a whole.46. although this route is not the main one [34].44). and S-methyl-acyl glucuronides.CHEMISTRY & BIODIVERSITY – Vol. methyl sulfoxides. an inhibitor of both ACE and neutral endopeptidase (also known as a vasopeptidase inhibitor) and a potential new drug for the treatment of hypertension [35]. In agreement with the caption to Fig. S-methylated metabolites accounted for a majority of a dose and demonstrated the significance of this pathway in the biotransformation of omapatrilat (4. In Figs.17.19.17 – 4. a closely related compound [36]. This drug undergoes marked S-methylation in humans to the inactive metabolite 4. It is interesting to note that these results do not appear to be isolated observations.43.42).

and rats. .54). Further medicinal examples are shown here. respectively.49.6) and reductive opening of the 1.2dithiolane ring (disulfide reduction) [37].53 and 4. dogs. which as a prodrug of 4. Individual differences in response and toxicity have been correlated with polymorphism in the TPMT gene and with the resulting differences in the extent of thiol methylation to yield metabolites 4. an endogenous cofactor as the (R)-enantiomer and an antioxidant drug as the racemate used in the treatment of diabetic polyneuropathy.2190 CHEMISTRY & BIODIVERSITY – Vol. most notably 6-mercaptopurine (4.50 as representative urinary metabolites in humans.52 shows similar variability in response and toxicity. see Chapt. 4. beginning with a-lipoic acid (4.18. and in organ transplants [33]. When administered orally to humans. yielding compounds 4.52) and 6-thioguanine (4.51). the drug was found to be extensively metabolized by boxidation (C2 chain shortening. 4. The latter reaction was followed by monoand di-S-methylation. The lower part of the Figure is dedicated to thiopurine MT substrates. and 4.48. 4. These drugs have been used in the treatment of leukemia and autoimmune disorders.47). 5 (2008) Fig.55. Another relevant drug is azathioprine (4.

56). The actual activation is the second step. there is a significant difference between clopidogrel and prasugrel. this step to the reactive 4. However. 5 (2008) 2191 Fig.58. Returning to thiol methyltransferase.61) in a reaction involving glutathione (see Chapt.62). but given that the overall reaction from 4. all remains to be clarified regarding the postulated CYP-catalyzed mechanism. The alternative. an agrochemical extensively used as a soil fumigant [39]. the active species which irreversibly antagonizes platelet ADP receptors via a covalent SÀS bridge.56). Our second example is that of metam (4. metabolic reaction is conjugation to Smethyl metam (4. its own metabolism is of pharmacological significance. This reaction appears as a detoxification although 4.62. In the case of prasugrel (4.58 is claimed to be catalyzed by CYPs. producing the S-methyl conjugate 4.7).19. Given the reactivity and activity of the thiol metabolite 4.60).57) [38]. 4.57 to 4.58 is a hydrolytic one. a recent analogue of the well-known platelet anti-aggregant clopidogrel. 4. which leads to a reactive thiol metabolite. we encounter prasugrel (4.CHEMISTRY & BIODIVERSITY – Vol. 4.59 and a cysteinyl disulfide conjugate (not shown). followed by tautomerization and double-bond rearrangement). and irreversible. This is a toxic dithiocarbamate which is reversibly transformed in animals into the reactive methyl isothiocyanate (4. 4. Like the latter. 4. and other metabolites are inhibitors of mitochondrial aldehyde dehydrogenase.60.58. . whose thiolactone formation is catalyzed respectively by cytochrome P450 and carboxylesterases (cleavage of the acetate ester.56 is a prodrug which must undergo a first activation step to a thiolactone (in this case metabolite 4.

since it exists in three valence states. However. Arsenic is present in many regions of the world where it diffuses from As-containing ores into water sources. some of the most toxic arsenicals are now believed to be methylated trivalent species [41]. Turning our attention to the metabolism of arsenic in humans and animals. namely metallic arsenic. At the level of tissues and organs. the inhalation of arsenic trioxide liberated by smelting causes lung cancer. and pentavalent arsenicals. since no thiol-As metabolite or intermediate is shown [43]. Thus. The contamination of the geosphere by arsenic causes severe health effects and cancer in exposed individuals. trivalent arsenicals. 4. the product of reduction of dimethylarsinate (4. arsenic is a carcinogen and may evoke severe inflammatory responses. Arsenic is also an occupational hazard for workers and populations associated with smelting of copper and other metals. the methylation of arsenic acids/ salts in organisms was considered to be a mechanism of detoxification. This Figure does not pretend to completeness. It is thus a natural contaminant to which hundreds of millions of humans are exposed through drinking water.2192 CHEMISTRY & BIODIVERSITY – Vol. and thiol conjugates (Chapt.21 is so arranged that reactions of reduction appear horizontally. whereas reactions of methylation appear vertically. In humans chroni- . most notably methylarsenicals of relevance here. 5 (2008) Figs.20 and 4. 4. A likely mechanism of DNA damage induced by methylated trivalent arsenicals involves the formation of reactive oxygen species.7). Furthermore. Fig. The manifold toxicity of arsenic is due in particular to its capacity to inhibit numerous functional proteins by binding avidly to thiol groups. it is now known that methylated arsenicals contribute significantly to arsenic toxicity and genotoxicity [40].67). and as organoarsenicals produced in the biosphere. Arsenic also exists as inorganic acids and salts. For many years.21. no more than dimethylarsine (Me2AsÀH). a fact that places arsenic redox reactions at the forefront of its toxicity [42]. The chemistry of arsenic is a rather complex one. 4.

20. The final steps to 4.20. and there are only few reports on the excretion of trimethylarsine oxide (4. 5 (2008) 2193 cally exposed to arsenic. dimethylarsinic acid) > dimethylarsinite (4.1) [8]. glutaredoxin:arsenate oxidoreductase (EC 1. An important finding is the fact that methylation occurs almost exclusively for trivalent arsenic species. the trivalent methylarsonite (4.67 (cacodylate.99. major methylated metabolites found in urine are 4.21.68. monomethylarsonic acid) > methylarsonite (4.4.65. an enzyme with absolute requirement for glutathione (GSH) and a member of the glutathione-S-transferases superfamily [48]. Comparable results were obtained in human and rat hepatocytes [45]. This step is catalyzed by at least two enzymes. and 4. the major (exclusive?) enzyme in As methylation is specific for trivalent species and is.66) is methylated by AS3MT to the pentavalent dimethylarsinate (4.66.64) is catalyzed by AS3MT and yields the pentavalent methylarsonate (4.20) [8] [46].70 are minor ones whose enzymology is unclear. Indeed.68.69) and trimethylarsine (4.64). known as arsenic(III) methyltransferase (Fig. Fig. In turn. dimethylarsinous acid) > methylarsonate (4.65).67). . monomethylarsonous acid) [44].CHEMISTRY & BIODIVERSITY – Vol.63) is reduced to arsenite (4. therefore.2).66) by glutathione:methylarsonate oxidoreductase (EC 1. pentavalent arsenicals must be reduced to trivalent arsenicals to allow their methylation [47 – 49]. arsenate (4. 4. 4. The latter is then reduced to methylarsonite (4.4. 4. The methylation of the trivalent arsenite (4. the major urinary metabolite of inorganic arsenic. In concrete terms.1) and arsenate:acceptor oxidoreductase (EC 1.69. Thus.20.70) in animals. The formation and excretion of trimethylated As species appears marginal.

The by-product of the reaction is adenosine 3’. Sulfonation reactions involve the nucleophilic attack on the S-atom by a OH group (in phenols. and is carried by a coenzyme.22. PAP). PAPS). and the group transferred is of medium molecular weight. .2194 CHEMISTRY & BIODIVERSITY – Vol. the sulfuric and phosphoric moieties are linked by an anhydride bond whose cleavage is exothermic and supplies enthalpy to the catalytic reaction. the formation of sulfates and sulfamates consists in a sulfonate group (À SOÀ ) being added to the substrate under 3 catalysis by a sulfotransferase. As shown here. This Chapter examines the formation of sulfates (RÀOÀSOÀ ) and 3 sulfamates (RÀNR’ÀSOÀ ) [5 – 8] [50]. This group is carried by the cofactor 3’-phosphoadenylyl sulfate (4. ionized. All criteria of conjugation are met in sulfonation reactions. since they are enzymatic.5’-bisphosphate (4. the reactions of sulfoconjugation are correctly designated as sulfonations rather than the common term of sulfations [51]. also known as 3’-phosphoadenosine 5’-phosphosulfate. the last Section of the Chapter will be dedicated to the (much rarer) reactions of xenobiotic phosphorylation. In PAPS. Given the analogies between sulfates and 3 phosphates. some sulfates are unstable under biological conditions and may undergo heterolytic cleavage to form electrophilic intermediates of considerable toxicological significance.72. 4. or a primary or secondary amino group. and hydroxylamides). hydroxylamines. alcohols.71. and highly hydrophilic. As we shall see. 5 (2008) Fig. As the group is a sulfonate.

the SULT2 family has a distinct substrate specificity for alcohol substrates.8. This family includes the subfamily SULT2A with 2A1 (alcohol/hydroxysteroid sulfotransferase. Sulfotransferases involved in the metabolism of small endogenous and exogenous molecules are soluble (cytosolic) enzymes. which is involved in amine sulfonation and seems to correspond with amine sulfotransferase (EC 2. the subfamily SULT1C with the enzymes 1C1 and 1C2.2.2. the subfamily SULT1B with the enzyme 1B1 (thyroid hormone sulfotransferase). EC 2. and which has been described as a brain sulfotransferase-like protein.4). also included are steroid sulfotransferase (EC 2. An important family is SULT3 with 3A1. 50 mammalian genes are known. and whose products are classified into families ( > 45% residue identity) and subfamilies ( > 60% residue identity) according to their degree of homology [8] [10] [52 – 54].1).14). In contrast.2.2). 1A2.2. the subfamily SULT2B with the two transcript variants 2B1a and 2B1b (EC 2.3). EC 2.8.8. they are now recognized as being encoded by a gene superfamily of which ca. EC 2.18). with some correspondence with EC 2.15) and cortisol sulfotransferase (glucocorticosteroid sulfotransferase.CHEMISTRY & BIODIVERSITY – Vol. and the subfamily SULT1E with 1E1 (estrogen sulfotransferase.8.8.8. and 1A3 (phenol ( ¼ aryl) sulfotransferases.2.8. Thus. human sulfotransferases include the SULT1A subfamily which contains the enzymes 1A1. .24. 4. Following major advances in molecular biology. All enzymes in the SULT1 family appear to have a marked preference for phenol substrates. and particularly hydroxysteroids.23 and 4. There is also a family SULT4 (with 4A1) whose substrate specificity is unknown.2.2. 5 (2008) 2195 Figs.

4. the alcohol sulfotransferases. Fig.2196 CHEMISTRY & BIODIVERSITY – Vol. it is wellestablished that the enzymes of greatest significance in the sulfonation of xenobiotics are the aryl sulfotransferases. and the homology-based nomenclatures. and the amine sulfotransferases. Nevertheless. a fact which prevents any strong and one-to-one correspondence between the classification of the Nomenclature Committee (NC) of the International Union of Biochemistry and Molecular Biology (IUBMB) [8]. 5 (2008) It is important to note that there are marked and condition-dependent overlaps in the substrate specificity of all these sulfotransferases. .24.

The nucleophilicity of the target O-atom in the substrate is increased by H-bond donation to His108. The sulfonate group in the center is still being weakly bound to the cofactor (PAPS in the process of becoming adenosine 3’. . the low-affinity. The lower part of the Figure states an important generalization about sulfonation reactions. but their velocity decreases as the concentration of available cofactor is rapidly depleted [58]. which are of high affinity and low capacity. The metaphor of a sprinter is a straightforward one. they have a fast initial turnover rate.CHEMISTRY & BIODIVERSITY – Vol. The upper part of this Figure shows a schematic and highly simplified depiction of the catalytic mechanism of sulfotransferases [55 – 57].5’bisphosphate) and has begun binding to the substrate (here a generic phenol).4). 5 (2008) 2197 Fig. Lys48 plays an important role in forming electrostatic bonds with the SO3 group and the phosphate moiety in PAP. The numbering is that of estrogen sulfotransferase.. 4. see Chapt. In concrete terms.e. and what the Figure shows is in fact the transition state. or other important residues. thereby stabilizing the former. the substrate. thereby acting directly or indirectly to decrease the free energy level of the transition state and stabilize it.25. The weak bonds (ionic and H-bonds) are represented by red broken lines of exaggerated length for clearer vision. especially when contrasted with that of a marathon runner (i. 4. highcapacity reactions of glucuronidation. the SO3 group. with excess electronic density on the three peripheral O-atoms and an electron deficiency on the S-atom. the transferred group is planar. At this stage. Other residues (among others Thr45 and Thr51) form H-bonds with the cofactor.

The optimized protein is colored using a color ramp. The Figure is also of general interest as it nicely illustrates the intricate manner by which the linear peptidic chain folds into a tertiary structure. the dark-grey model on the left of the picture). A more realistic view of the proximity between cofactor and substrate in the active site of sulfotransferase is presented. and the cofactor is a PAP analog (the violet model on the right. The structure was completed by adding the H-atoms and minimized using VEGA. 5 (2008) Fig. courtesy of Dr. 4. Giulio Vistoli.11. .18 in Fig. A single monomer was used to build the image. University of Milan). and the ligands are depicted as Van der Waals space-filling models. the image was generated by VMD and rendered by POVRay [59]. The structure of SULT1A1 crystallized with the cofactor and a phenolic substrate was retrieved from the Protein Data Bank (PDB ID: 2D06) [56]. 4. Secondary structure elements are represented as coils for helices and arrows for b-strands. The substrate is estradiol (4.26.2198 CHEMISTRY & BIODIVERSITY – Vol. from blue (N-terminus) to red (C-terminus).

73) was used to this end. yielding useful structure – affinity relationships.74) has a high substrate affinity for SULT1A1 and is a poor substrate of other SULT1 enzymes [60].1’-biphenyl (4.5.76).4) clearly predominating over formation of the monoO-sulfate (4. namely bisphenol A (4. Other examples of preferred glucuronidation over sulfonation will be presented later. In contrast.1’-biphenyls such as 2’. 5 (2008) 2199 Fig. phenolic metabolites of polyhalogenated 1.g. with formation of the mono-Oglucuronide (4. since it is also a substrate of 1B1 and other SULTs. but recent results dispute its high selectivity. 2-aminophenol (4.78.77 were found to be substrates of human hydroxysteroid SULT2A1.5.27.. see Chapt. R ¼ H) and tetrabromobisphenol A (4. it was identified as a non-substrate inhibitor of the enzyme [62]. Their conjugation may be hypothesized to be a route of detoxification. 3.77). making them potential competitors of the sulfonation of endogenous steroids. . e. As for 4. For years.75. we encounter two simple compounds used as probes of the major human hepatic sulfotransferase 1A1.80). whereas the former is an endocrine disruptor and one of the highest volume chemicals produced worldwide.79.4’-tetrachloro-4-hydroxy-1. A large number of other phenols were examined for their substrate behavior toward aryl sulfotransferases [61]. Beginning with model and xenobiotic phenols. 4-nitrophenol (4. Our last examples here are two xenobiotics of toxicological concern.1’-biphenyl (4.2’. and 3.1’-biphenyl (4. Other phenols stand out for their environmental and toxicological significance. being used as a monomer and in many plastic consumer products such as toys and water containers [62]. Phenols 4. 4. These properties are a cause of concern as they suggest an interference with hormone regulation.75).78. The latter is a flame retardant with demonstrated cytotoxicity. R ¼ Br).CHEMISTRY & BIODIVERSITY – Vol. 4.5’-dichloro-4hydroxy-1.2’-trichloro-4-hydroxy-1.76 and 4.

see Chapt. 4’-O-sulfate 4. 5 (2008) Fig. Dopamine of endogenous or exogenous origin is a good substrate of sulfotransferases and particularly SULT1A3 [63]. a polyphenol present in grapes and wine. dietary polyphenols [52] [64]. An example is provided here with (E)resveratrol (4.84. and 3. the major metabolites were the 3-O-glucuronide and the 4’-O-glucuronide (see Fig. hence. Incubations with recombinant enzymes uncovered the enzyme selectivities shown. 4. and the active metabolite of the same l-DOPA used as a major anti-Parkinsonian drug. In PAPS-fortified human liver cytosol (where no glucuronyltransferases are present.83). 4. The high regiospecificity of the enzyme for the meta-OH group (to yield 4. and dopamine O-sulfate.2200 CHEMISTRY & BIODIVERSITY – Vol. see also Fig. .85.10) is of particular interest. Dopamine (4.11.82. 4. the Figure also shows two other routes of inactivation of dopamine.4). namely COMTcatalyzed O-methylation and MAO-catalyzed oxidative deamination. in particular.42). 90% of all circulating dopamine in humans. with sulfonation being its major route of inactivation. Many natural and endogenous compounds are substrates of sulfotransferases. the major metabolites were the 3-O-sulfate 4.28.9). For the sake of a broader vision. 4. its name of monoaminesulfating phenol transferase (see Fig. 4. 4. In vivo in rodents and in rat hepatocytes. Numerous xenobiotics of plant origin are also substrates of sulfotransferases.84 and the 3-O-glucuronide [65].23).4’-O-disulfate. three sulfates were produced.81 and 4.81) is also remarkable. accounting for ca. The high selectivity of dopamine toward SULT1A3 is explained by the presence in the catalytic site of a carboxylate group forming an ionic bond with the ammonium group of dopamine and related amines. and known to be endowed with cardioprotective activity. In human hepatocytes. being both an essential neurotransmitter produced by decarboxylation of the amino acid l-DOPA (4. namely the 3-O-sulfate 4.

4..88) [66]. In vitro studies using human intestinal cytosol and recombinant human SULT1A3 have confirmed the effective sulfonation of several b2receptor agonists including isoprenaline (4. namely b2-receptor agonists used in the treatment of asthma. Traxoprodil (4. their affinity proved markedly structure-dependent.CHEMISTRY & BIODIVERSITY – Vol. Here. see Part 6). There was also a modest stereoselectivity such that the inactive (þ)-(S)enantiomers were somewhat better substrates. but direct conjugation by O-glucuronidation and O-sulfonation was the major metabolic pathway in poor metabolizers (i. both the drug and this metabolite form the respective sulfate esters 4. often in competition with glucuronidation (Chapt. 4.e. 4. we present three such analogues.86. with isoprenaline being the best substrate. While all drugs examined had comparable Vmax values. Interestingly.14). persons with defective CYP2D6 activity. 4. 4. The previous Figure mentioned the role of monoamine-sulfating phenol transferase (SULT1A3) in the conjugation of xenobiotic analogues of dopamine. and formoterol (4.10) offers a nice illustration of sulfonation occurring for both the drug and one of its metabolites [19].89 and 4. a number of phenolic drug metabolites undergo sulfoconjugation.4). .13. isoproterenol).90 along with the O-glucuronides. see also Fig. terbutaline (4.13 undergoes CYP-catalyzed ring oxidation followed by COMTcatalyzed O-methylation to form 3’-methoxytraxoprodil (4. Ring oxidation was the major route in most subjects.2. Besides drugs. 5 (2008) 2201 Fig.87). As a rule. As discussed in Chapt. such drugs are quite hydrophilic and undergo substantial presystemic metabolism mainly by sulfonation.29.

2202 CHEMISTRY & BIODIVERSITY – Vol. 4.23). the sulfonation of primary alcohols is distinctly faster than that of secondary alcohols. However.91).97.5-dichlorophenyl)succinimide (4. since it is able to react with other endogenous nucleophiles in addition to GSH. the presence of the adjacent ÀCH2 À group allows intramolecular deactivation to the unsaturated maleimide analogue 4. This is well illustrated with three isomeric C7 alcohols. hence the blue arrows) either directly or via the carbocation to form the conjugate 4. Like aryl sulfates. The sulfonation of alcohols is also of metabolic significance. some higher-molecular-weight sulfates may also be toxic. with that of tertiary alcohols being minute. see Chapt. The influence of substrate structure on their sulfonation has led to some interesting generalizations [67]. trans-4-methylcyclohexanol (4.93). Thus.96 [68].30. but the involvement of other SULTs is sometimes documented. 5 (2008) Fig. Its oxidation and sulfonation in rats yielded the sulfate 4. Thus. The latter is believed to be the ultimate nephrotoxin. alkyl sulfates are sensitive to enzymatic and chemical hydrolysis. Interestingly. namely cyclohexylmethanol (4.92).95 which reacted very rapidly with glutathione (GSH. given that sulfates of low-molecularweight alcohols such as methyl sulfate and ethyl sulfate are known alkylating agents. 4.7. with the added feature that the resulting alkyl or cycloalkyl sulfate esters may be reactive. Another important rule is the fact that low-molecular-weight alcohols (less than six Catoms) are poor substrates. 4. . and 1-methylcyclohexanol (4. several alkyl sulfates are known to undergo proton-catalyzed heterolytic cleavage as discussed below. the agricultural fungicide N-(3. The enzymes involved in alcohol sulfonation are mainly alcohol/ hydroxysteroid sulfotransferases (see Fig. This is fortunate.94. NDPS) is a known nephrotoxin. and as illustrated here and in the next Figure. in addition.

e. 5 (2008) 2203 Fig. Besides being inactivated by glutathione.98)). as exemplified here [70 – 72].99. produces the sulfate 4. isosafrole. see next Figure) of sulfates [69]. 4. The molecular features favoring heterolytic cleavage are partly known.5-dichlorophenyl)succinimide (4. and analogues [69]). .7) to form 4.g. Chapt. Such delocalization can be caused by an adjacent unsaturated system such as a carbonyl group (as in N-(3. followed by sulfonation catalyzed by hydroxysteroid sulfotransferases. it also undergoes a heterolytic CÀOSOÀ 3 cleavage to generate the carbocation.101). delocalization of the positive charge) certainly contributes to the stabilization of the carbocation and hence to an increased likelihood of heterolytic cleavage. the stability of the carbocation appearing as a significant determinant [69].. an aromatic system (e. the carbocation forms adducts with DNA. This reactive conjugate can be detoxified by a direct substitution catalyzed by GSHtransferases (GST.94)).100.31.CHEMISTRY & BIODIVERSITY – Vol.. This reaction is already 3 apparent in the previous Figure. CYP-Catalyzed hydroxylation of polycyclic methylarenes (here 5-methylchrysene (4. Resonance (i. particularly at the nucleophilic NH2 group of adenine (see 4. Reactive electrophiles are formed by the heterolytic CÀOSOÀ cleavage 3 (and NÀOSOÀ cleavage. or an allylbenzene structure (as in safrole. 4. methylarenes). and it is particularly worrying for genotoxic and carcinogenic polycyclic arylmethanols.

9) and SULT2B1 [70] [72] [73]. for example. three metabolites were found to be substrates of sulfotransferases. Most known substrates in this broad class are hydroxylamines and hydroxylamides. 5 (2008) Fig. The fate of 4. since sulfonation of the NHOH group leads to a labile sulfate.107).32. namely 2-nitrobenzyl alcohol (4.108.2204 CHEMISTRY & BIODIVERSITY – Vol. 4. In male rats. A number of N-oxygenated compounds are also known substrates of sulfotransferases. 4.106. 2-aminobenzyl alcohol (4. 4.2.104 is plainly similar to the examples discussed in Figs.103). which undergoes proton-catalyzed heterolytic cleavage to a nitrenium cation stabilized by resonance as shown.102) [74]. thereby forming O-sulfates (also known as N.107 is of relevance here. namely 2nitrotoluene (4. The case of 4. . with the sulfate ester 4. as exemplified here with a simple and interesting model compound and industrial chemical.104).O-sulfates).31.105 forming a carbocation and the glutathione conjugate 4. and 2-(hydroxyamino)benzyl alcohol (4. aryl sulfotransferase IV (EC 2. The sulfonation of the latter two was concluded to account for covalent DNA binding in liver.8.30 and 4.

This compound served as a model of amidoximes. and the actual active form responsible for its therapeutic effects is its stable.110 reacts as shown to form a highly electrophilic nitrenium ion (whose detoxification is shown in Fig.4) might be a route of toxification. A classical example is that of N-(9H-fluoren-2-yl)-Nhydroxyacetamide (4. and their sulfate esters are characterized by a great reactivity in aqueous media [75a]. some of which are of interest as bioreductive prodrugs (see Part 5) of medicinal amidines.115).116 [77]. However. Many conditions are.2.O-sulfate 4. 4. and/or necrotic sulfate conjugates. 4. carcinogenic.114 (see Chapt. 4. Despite the potential toxicity of sulfates of hydroxylamines and hydroxylamides.112).112 and benzamidine (4. Hydroxylamides (also known as hydroxamic acids) might be even more carcinogenic than hydroxylamines following their toxification by sulfonation. Its O-sulfate 4. An intriguing and rare reaction of conjugation occurs for minoxidil (4. fulfilled for aromatic amines and their N-acetylated conjugates (see Chapt. one should not conclude that O-sulfonation of N-oxygenated compounds always implies toxification. 4. there were fears that the conjugation of amidoximes leading to the Osulfate 4. a hypotensive agent also favoring hair growth. and a good substrate to rat liver aryl sulfotransferase IV and human liver SULT2B1 [75]. . they are sulfonated faster than the corresponding hydroxylamines by aryl sulfotransferase IV (EC 2. This is demonstrated with amidoximes such as benzamidoxime (4. a metabolite resulting from the CYP-catalyzed N-hydroxylation of N-(9H-fluoren-2-yl)acetamide.9). The redox equilibrium between 4.118).111) markedly favors reduction.109).8.CHEMISTRY & BIODIVERSITY – Vol. This drug is an Noxide. zwitterionic N. 5 (2008) 2205 Fig. Nevertheless. Indeed.113 and the predominant O-glucuronide 4.5) to form mutagenic.33. therefore. the two conjugates proved chemically stable and devoid of mutagenic effects [76].

4-tetrahydroquinoline (4. One medicinally relevant example is that of trovafloxacin (4.2. indicating its stability against biodegradation by the gut microflora (see also Sect.121) and octanamine (4.3.118) which accounted for ca. perhaps depending on the enzyme involved [80].5. This may not necessarily be the rule for lipophilic medicinal amines such as desipramine. Examples of good substrates include the weakly basic (pKa 4 – 5) aromatic amines naphthalen-2-amine (4. In contrast to unstable hydroxylamine sulfates. 30%) and the N-glucuronide 4.2. Chapt. but only when incubated at basic pH where they are in their neutral form.4-tetrahydroisoquinoline (4.123).120). Human volunteers administered the drug excreted it partly unchanged and partly as three major conjugates. a quinolone antibacterial agent. 10% of the dose and was excreted fecally. The sulfonation of a non-basic N-atom is also documented with luminol (4. a luminescent reagent used in forensic toxicology. namely the sulfamate 4.125 (ca. The involvement of amine sulfotransferase (EC 2.e. The basic (pKa 8 – 10) amines 1. 4.8. 5 (2008) Fig. and whose fate was investigated in rats to search for possible toxic metabolites [81].124 (ca. and arylamines can all yield sulfamates in the presence of human sulfotransferases. 4.3) remains to be better understood. 60%.2..34. in particular SULT2A1 [79].122) are also good substrates. one of which was the sulfamate (4.117). . and primary and secondary alkylamines. especially in humans. 4. A number of alicyclic amines. only two inert metabolites were found.4).119) and 1. sulfamates) are obtained upon N-sulfoconjugation of amines.3.2206 CHEMISTRY & BIODIVERSITY – Vol. Besides the parent compound ( < 5% of a dose). significantly more stable conjugates (i.2) [78].

21).CHEMISTRY & BIODIVERSITY – Vol.7).8).7.7. nucleoside phosphotransferase (EC 2. nucleoside-phosphate kinase (EC deoxyadenosine kinase (EC 2.130) itself is prochiral (it bears two enantiotopic CH2OH groups). The agent is phosphorylated in rats and humans to the active monophosphate 4. creatine kinase (EC 2.7. .7. (deoxy)nucleoside-phosphate kinase (EC 2. 5 (2008) 2207 Fig. the diphosphate 4. Such reactions are sometimes.7. They are known or postulated to be catalyzed by some among the many phosphotransferases (EC 2.4.20).1.131.13). adenosine kinase (EC 2. The monophosphate 4. uridine kinase (2.e.1. which is also the only active one.2).7. Indeed.7.127 was the predominant compound.128 and triphosphate 4.7.14) [8]. phosphorylation is an essential metabolic step in the bioactivation of these agents.76). a novel immunomodulator used in transplantations and to treat autoimmune diseases [84]. labeled as anabolic (i. adenylate kinase (EC 2.1.7. nucleosidediphosphate kinase ( deoxycytidine kinase (EC 2.1.3).131).77).126. Indeed. and correctly. azidodeoxythymidine).3.74).1. Phosphate conjugates are rare compared to sulfates but are of primary significance in the metabolism of anticancer and antiviral agents impacting on endogenous nucleotides. and its enzymatic phosphorylation yields exclusively the enantiomer of (S)-configuration (see 4. for example.129 were present in minor and comparable amounts. di-.6).7. and numerous studies document their stepwise phosphorylation to mono-. An example is afforded by the well-known anti-HIV agent zidovudine (4. FTY720 (4. The reaction is catalyzed by sphingosine kinases and is highly productenantioselective. biosynthetic) ones [82]. 4.7. guanylate kinase (EC 2.. and triphosphates. AZT.4.7. The unexpected activity of phosphotransferases toward xenobiotic substrates is also seen with FTY720 (4. pyruvate kinase (2. The concentrations of its phosphate anabolites were measured in the peripheral blood mononuclear cells of AIDS patients treated with the drug [83].4.48). thymidine kinase (2.35.130).7. and pyrimidine nucleoside monophosphate kinase (2.

4. The pKa of glucuronic acid is 3. we begin with the biochemistry of glucuronidation. 5 (2008) Fig.9 – 3.2208 CHEMISTRY & BIODIVERSITY – Vol. leaving the presentation of the enzymes proper (UGTs) for later Figures. UDPGA) to the substrate. Together with glutathione transferases (Chapt. 5 g are synthesized daily in the adult human body. a common characteristic they share despite their great chemical variety. implying nearcomplete ionization in the physiological pH range [86]. 4.133. but the products of conjugation are b-glucuronides 4. Glucuronosyltransferases are low-affinity enzymes compared to sulfotransferases. This cofactor is produced endogenously by the C(6) oxidation of UDP-a-d-glucose. and it is recognized that ca. . all target sites in substrates are nucleophiles.36.132) exists in the (1a)-configuration.25). and quantitatively for the vast number and diversity of their substrates [5 – 10] [50] [85]. Indeed. hence the high capacity of this metabolic route. This is due to the mechanism of the reaction being a nucleophilic substitution with inversion of configuration.1. Here. This is explained in qualitative terms by the diversity of functional groups to which glucuronic acid can be coupled. An important feature of glucuronides is their acidity. Glucuronidation consists in a molecule of glucuronic acid being transferred from the cofactor uridine-5’-diphospho-a-d-glucuronic acid (4. Glucuronic acid in UDPGA (4.0. 4.7). glucuronosyltransferases count as the most significant conjugating enzymes in xenobiotic metabolism.132. and that of O-glucuronides in the range 2. which implies that sulfonation is usually faster than glucuronidation at low substrate doses or concentrations (see Fig.

5 (2008) 2209 Fig.e. First. they are thus zwitterions [87]. of the pyridine type). hydroxylamines. . The previous Figure contains a summary of the functional groups being potential substrates for UDP-glucuronyltransferases. or aromatic (i.37. A few strongly acidic enolic acids are known to form Cglucuronides. namely O-. and hydroxylamides. S-. there are the OH groups in phenols. Another group of N-glucuronides are formed from tertiary amines. which form O-glucuronides. saturated heterocyclic. An important class of O-glucuronides are also the acyl glucuronides formed from carboxylic acids (shown here) and carbamic acids (RR’NÀCOOH. see Chapt. 4. the products of glucuronidation are grouped into four main classes. be they aliphatic. The N-glucuronides are generated from primary and secondary aliphatic or aromatic amines. alcohols.CHEMISTRY & BIODIVERSITY – Vol. These N-glucuronides are special in the sense that they contain a permanent positive charge in addition to the negative charge of the carboxylate. and C-glucuronides. Thiols and thioacids can lead to S-glucuronides. amides and sulfonamides (all of which are represented as RR’NH in the Figure).. N-. saturated secondary heterocyclic amines. In summary. 4.8). This overview is presented here in greater detail and shows the electron-rich target sites of glucuronidation. depending on the site of attachment of the C(1)-atom in glucuronic acid.

4.e. epiandrosterone). stomach. and bilirubin (a waste product of hemoglobin) whose detoxification by UGTs is of major significance in humans [92]. 2B4. These enzymes are part of the glycosyltransferases. UGT2. androsterone. 4. mutations in 1A1 cause hereditary unconjugated hyperbilirubinemia (i. 4. Thus. A number of polymorphisms have been reported in the UGT1 and -2 families.40 with the evolutionary relationship of human UGTs in families 1 and 2 [88] [89]. and 2B17. Endogenous substrates include a variety of androgens (testosterone. 5 (2008) Figs. This is complemented in Fig. 2B15. reproductive organs (the mammary glands.39 together with the major classes of their substrates [96] [97]. The enzymes catalyzing the highly diverse reactions of glucuronidation are known as UDP-glucuronosyltransferases (UDP-glucuronyltransferases.38 – 4. 2B7. gastrointestinal tract (esophagus. small intestine. and the skin. and prostate). 4. Crigler – Najar and Gilbert syndromes). chenodeoxycholic acid). The human UGTs known to metabolize xenobiotics are the products of (to date) four gene families (UGT1. e. 2B10. UGTs) and consist in a number of proteins coded by genes of the UGT superfamily [6 – 8] [88 – 94]. UGT3. and UGT8). Their tissular distribution is quite broad. and colon). with noteworthy organs being the liver and bile ducts. estrogens (bestradiol. testes.9 that UGTs are also able to catalyze conjugations with glucose and a few other hexoses [95]. estriol) and gestagens (17a-hydroxyprogesterone). .2210 CHEMISTRY & BIODIVERSITY – Vol.. deoxycholic acid.g. These are membranebound enzymes found in the endoplasmic reticulum.4.. biliary acids (lithocholic acid. and we shall see in Sect.40. Other polymorphisms are known in. kidneys. A list of human UGTs is shown in Fig. UGT1A3 to 1A10.

5 (2008) 2211 Fig. 4.40.39. 4. Fig.CHEMISTRY & BIODIVERSITY – Vol. .

The predicted backbone was completed by adding the side chains and the H-atoms using VEGA and was joined to the experimental C-terminus by superimposing the common residues. University of Milan. the ligands were docked manually optimizing their known interactions with pivotal residues of the UGT2B7. The structure of the enzyme was generated combining the C-terminal domain (Ala285 – Ser446) whose structure was experimentally resolved (PDB 2O6L [98]) with the model of N-terminus (Gly24 – Pro284. the N-terminal domain was modeled by Fugue exploiting its homology with the macrolide glycosyltransferase from Streptomyces antibioticus (PDB 2IYF). In detail. 4.2212 CHEMISTRY & BIODIVERSITY – Vol. 4. see also Fig. the putative signal peptide from Met1 to Cys23 was discarded) which was generated by homology techniques. 5 (2008) Fig.41. . The ligands are represented as Van der Waals spheres (green for morphine and cyan for the cofactor UDPGA). With the apoprotein structure minimized. The resulting structure was finally minimized to optimize the relative position of the bound ligands (courtesy of Dr. the tertiary complex was generated by docking first UDPGA. Specifically. The Figure shows the ribbon structure of UGT2B7 complexed with UDPglucuronic acid and morphine. Giulio Vistoli.26). The ribbon is colored using a ramp of color from blue for N-terminus to red for C-terminus. followed by morphine.

Our overview of UGTs substrates begins with phenols. than 4’-O-glucuronide. 5 (2008) 2213 Fig. 4. herbivores and omnivores have been exposed to such nonnutritious compounds throughout their evolutionary history.CHEMISTRY & BIODIVERSITY – Vol. and quite logically with plant phenols. 4. Baicalin (4.136. but only at very low substrate concentrations [101]. lipid-lowering. UGT1A1 was the most efficient. cardioprotective and chemopreventive activities. in the presence of human or rat liver or intestinal microsomes. Both sulfonation (see Fig.137) bears OH groups on its B-ring only. Its regioselective glucuronidation is also markedly dependent on enzyme and tissue. 7-O-Glucuronidation was catalyzed by the human enzymes 1A9 > 1A8 > 1A3 > 2B15 > 1A7. Quercetin (4. SULTs and UGTs among them [99]. .and 3’-positions seemingly emerging as preferred over the 7. (E)-resveratrol (4. Thus.83) is found in a variety of plant sources.and 4’-positions [102].36). Overall. which explains the evolution of detoxifying enzymes to facilitate their elimination. their relative importance depending on dose (i. the latter were markedly more active than the former [100]. 4. with no reaction detected at the 5-OH group. 4. and is known for its antioxidant. Indeed. with the 3. The two Oglucuronides of (E)-resveratrol (4. the rates of O-glucuronidation were higher at 7-OH than at 6-OH. most notably grapes. Human liver microsomes produced more 3-O-glucuronide. 4.e.138) is more complex as it contains four phenolic and one phenol-like OH group. Two further examples of product regioselective glucuronidation are seen with bioflavonoids such as baicalin and quercetin. a product regioselectivity reversed in human intestine microsomes.83) are shown here together with the human UGTs that catalyze their formation. low doses allow for higher degrees of sulfonation.42.135.28) and glucuronidation are significant reactions in humans.. see Fig.

2214 CHEMISTRY & BIODIVERSITY – Vol. its main metabolite is the O-glucuronide 4.6-O-diglucuronide and morphine 3-O-sulfate (not shown here) [103].140) and morphine 6-Oglucuronide (4. 4. these reactions are catalyzed mainly by UGT2B7. and that both the 3-Oand the 6-O-glucuronides accumulate in the serum of renally impaired patients chronically treated with morphine. Lower amounts of two other conjugates have also been found in a number of species. 1A6. At therapeutic doses. 50 – 60% of total urinary metabolites). In most laboratory species and in humans. In humans. namely the 3. 30 – 40% of total urinary metabolites).142). The presence of UGT2B7 in human central nervous system provides further evidence for the therapeutic contribution of its 6-Oglucuronide to the analgesic activity of morphine.141). At hepatotoxic doses. An apt transition between natural products and medicinal compounds is provided by morphine (4. 5 (2008) Fig. Another important example is that of paracetamol (4. Of great clinical significance is the fact that morphine 6-Oglucuronide is a highly active agonist at the opiate m-receptor. This major medicine contains a phenol function (the 3OH group) and a secondary alcohol function (the 6-OH group). A further but somewhat less important conjugate is the O-sulfate 4. respectively.143 (ca. . CYP-Catalyzed oxidation (a route of toxification) represents less than 5% of a dose [104] (see also Parts 2 and 3). with oxidation also increasing (7 – 15%). the former is produced in higher amounts than the latter. yielding morphine 3-O-glucuronide (4. A number of UGTs can glucuronidate paracetamol. and 1A9.43. the sulfonation capacity is exceeded and glucuronidation predominates (2/3 to 3/4 of metabolites).139).144 (ca. Both groups are glucuroconjugated. a nonprescription drug widely used for its mild analgesic and antipyretic properties. the most active ones being UGT1A1.

145). an inhibitor of the intestinal absorption of exogenous cholesterol used in the treatment of primary hypercholesterolemia.145) is interesting as a substrate of UGTs in that it contains two target sites. given that the pharmacological target of 4.5 mg [106]. alone or in combination with cholesterol-lowering drugs acting by another mechanism (e. and 2B15) is produced in much larger proportions than the benzylic metabolite 4. the phenolic metabolite is the main circulating metabolite in human plasma. A number of other drugs have their therapeutic effects prolonged by enterohepatic cycling. In this Figure. and that its glucuronides are also active. statins). Both conjugates are excreted mainly in bile and undergo enterohepatic cycling. The phenolic glucuronide 4.145 is an intestinal cholesterol transporter.146 (formed by UGT1A1. as nicely illustrated in a pharmacokinetic study of estradiol in postmenopausal women administered a single oral dose of 1.44. .CHEMISTRY & BIODIVERSITY – Vol. This phenomenon is clinically relevant.g.. Ezetimibe (4. 5 (2008) 2215 Fig. human jejunum microsomes produced the two O-glucuronides in similar amounts. namely a phenol and an alcohol group. In contrast. In fact. Another medicinal example of glucuronidation is ezetimibe (4. it is also and by far the major in vitro conjugate in human liver microsomes. 1A3.147 (formed by UGT2B7) [105].148). 4.18) in redox equilibrium with its metabolite estrone (4. we simply show the structure of estradiol (4.

However. It is known from numerous literature data that large hydrophilic anions are actively secreted in the bile ducts as substrates of organic anion transporters whose primary substrates are biliary acids produced in the liver. a second absorption phase rapidly kicked in and resulted in sustained levels of estradiol for 24 h and more.18) and its metabolite estrone (4. Estrone levels were markedly higher than estradiol levels (cmax % 130 vs. 450 – 500 Da in humans and 350 – 400 Da in rats. And the aglycone liberated by hydrolysis must obviously be absorbable by the intestinal wall. 30 ng/l). biliary excretion is a necessary but not sufficient condition for enterohepatic cycling. This Figure continues and complements the previous one by illustrating some of the pharmacokinetic results obtained in the clinical study under discussion [106]. . The time profile of estradiol shows a first phase of absorption – elimination with an approximate half-life of 2 h. 4. 5 (2008) Fig. The serum concentration curve of the metabolically produced estrone showed enterohepatic absorption phases after ca. since the glucuronide must also be hydrolyzable by bacterial and/or intestinal glucuronidases [6] [107] [108]. the biliary excretion occurs above a threshold of ca.2216 CHEMISTRY & BIODIVERSITY – Vol. By virtue of their negative charge.45. Both estradiol (4.148) were extensively Oglucuronidated and underwent enterohepatic cycling. An experimental proof of this phenomenon can be seen in the time profile of the serum concentrations of both hormones. extending its half-life from 4 h after the first absorption phase to 11 h. 24 and 50 h. As a rule of thumb. the glucuronides of sufficiently large aglycones fulfill the physicochemical conditions for biliary excretion. However.

whereas linalool is a tertiary one. Interestingly. to a lesser extent. To follow the same sequence as Chapt. This reaction was catalyzed by UGT1A8 and. a point we shall take up in the next Figure. .3. However.150 was catalyzed by human UGT2B17. we devote a Figure to natural alcohols.153 [110]. recent studies using powerful new technologies have identified a novel class of glucuronides resulting from the sequential glucuronidation of a single OH group. Alcohols of plant origin are numerous and. have been consumed by herbivores for hundreds of millions of years.3). Incubation of the monoglucuronide resulted in a low but measurable production of the diglucuronide 4.CHEMISTRY & BIODIVERSITY – Vol. by 1A1 and 1A9. As examples. UGT1A8 was the only human UGT capable of producing the diglucuronide from 5a-dihydrotestosterone.149). explaining for a good part the evolution of the UGT gene superfamily. like natural phenols. Alcohols can be glucuronidated with relative ease. ( À )-borneol and ( Æ )-linalool are two terpenoid alcohols known to form the respective glucuronides 4.46. These endogenous compounds and their synthetic analogues are well known substrates of UGTs in humans and animals. 4. 5 (2008) 2217 Fig.3.151 having the two glucuronyl moieties linked at the 1’’ ! 2’ position. Note that borneol is a secondary alcohol. beginning with steroidal hormones [109]. 2B15. This is illustrated here with 5a-dihydrotestosterone (4. 1A8. Its glucuronidation at the 17b-OH group to yield the monoglucuronide 4. and 1A4 in decreasing order. 4. producing O-glucuronides whose chemical stability contrasts with the reactivity of O-sulfates (see Sect. 4.152 and 4. the active metabolite of testosterone.

a reaction catalyzed by human UGT2B7 [112b].157 which was a major metabolite.1-dimethylpropyl methyl ether (4. The lower part of the Figure shows relevant metabolites of 1. and mainly at the secondary OH group at C(3) (with some glucuronidation at the 4-CH2OH group) in rat microsomes [112a].156 led to 1.3-diol (4. Major urinary metabolites were 2-methylbutane-2. such a result cannot be generalized given its dependence on species and isozymes. except for 4. When chemically diverse alcohol groups are present in the same molecule.158.155) was glucuronidated at the secondary OH group at C(2) in dog microsomes.2218 CHEMISTRY & BIODIVERSITY – Vol. 5 (2008) Fig. Comparable urinary ratios were seen in a single human volunteer. Fig. tenfold preference for the former [111].126. the glucuronide (4. The results in humans are consistent with the high O-glucuronidation of AZT (4. This Figure addresses the issue of chemoselectivity and regioselectivity in reactions of glucuronidation (see also Part 1 [1] [4]).159) of the tertiary alcohol 4.35) at its corresponding CH2OH position.160 which was a minor. and the 3-O-glucuronide 4. Chemoselectivity implies the discrimination between chemically diverse target groups. independently of their presence in the same or different compounds. Human microsomes produced both the 3-Oglucuronide (at the primary OH group) and the 1-O-glucuronide (at the secondary OH group). 4.156) in rats [113]. secondary and tertiary alcoholic groups.160 of 2-methylbutane-2.157). at the primary 4-CH2OH group in human microsomes. as seen with the antibiotic chloramphenicol (4. Indeed.158) resulting from further oxidation.47. chemoselectivity overlaps partly with regioselectivity. Thus. . However.154). CYP-catalyzed oxidation of 4.e. and the glucuronide 4.. i. the antithrombotic agent AZ11939714 (4. a minor urinary metabolite. at its secondary alcoholic group).3-diol (4. which affords a further example of the glucuronidation of a tertiary alcohol.1-dimethylpropan-1-ol (4. 4. with a ca.157. here between primary.

The case of oxazepam (4.and (R)-propranolol. whereas UGT2B15 is specific for (S)-oxazepam [116]. .161) is a chiral drug existing as two stable forms. In humans administered racemic propranolol.CHEMISTRY & BIODIVERSITY – Vol. and UGT2B7. 4. since UGT1A9 and 2B7 glucuronidate (R)-oxazepam. 5 (2008) 2219 Fig. Because we prefer to use the term selectivity for reactions and specificity for enzymes [1] [4]. and more generally for one stereoisomer over the other(s). the eutomer) and the inactive (R)-enantiomer (i.162) is both similar to and different from that of propranolol. Because d-glucuronic acid is itself a chiral. plasma and urinary levels of (S)-propranolol glucuronide are higher than those of (R)-propranolol glucuronide. what is presented here are examples of the enantiospecificity of UGTs..e. Yet. Another type of selectivity is seen in some glucuronidation reactions as well as in various other phase I and phase II metabolic reactions. which show a marked enantiospecificity for (S).e. UGTs acting on this substrate do show enantiospecificity. whose preference for (R)-propranolol is minute [114]. such that the drug racemizes with an estimated halflife of 1 – 4 min under physiological conditions of pH and temperature [115]. the overall reaction of propranolol glucuronidation is substrate-enantioselective. the glucuronidation of propranolol (4. the active (S)-enantiomer (i. The origin of this selectivity has now been ascribed (at least in part) to three UGTs. namely UGT1A9 and 1A10.161) yields two diastereoisomeric glucuronides.. In other words. respectively. the distomer). namely the preference of some UGTs for one enantiomer over the other.48. The b-blocker propranolol (4. enantiomerically pure compound. The stereogenic center in this compound is a highly unstable one.

An example is found in the metabolism of the candidate hypoglycemic agent 9-[(1S. but rat liver microsomes do not. explaining the faster clearance of the (S)-isomer in humans. Hydroxylamines and hydroxylamides may also form O-glucuronides. It is also interesting to note that all results reported in this study are consistent with a good chemical stability of the O-glucuronide 4.166).2220 CHEMISTRY & BIODIVERSITY – Vol. .49.163) [117].3. four times higher than that of its (R)-enantiomer. It is an unusual drug in that it contains a hydroxylamido group which proved to be a major target of human UGT [118]. Interestingly.164.2R)-2-fluoro-1-methylpropyl]-2-methoxy-6-(piperazin-1yl)purine (4. When administered to monkeys. Indeed.165. The possibility of the metabolite being an Nglucuronide was conclusively excluded. an inhibitor of 5-lipoxygenase. 4. 4. has value in the treatment of asthma and other inflammatory pathologies. since the glucuronidation rate of (S)-zileuton in human liver microsomes is ca.165. Substrate enantioselectivity is also documented. 5 (2008) Fig.167 in human urine. ca. 75% of a dose are recovered as the O-glucuronide 4. more than half of a dose was recovered as a compound found to be the O-glucuronide. Zileuton (4. a few drugs and a number of aromatic amines are known to be N-hydroxylated and then O-glucuronidated. 4. in contrast to some O-sulfates of hydroxylamines and hydroxylamides as discussed in Sect. of the Nhydroxylated metabolite 4. Thus.4. human and monkey liver microsomes catalyze the formation of this metabolite.

particularly to NH2 groups in blood and tissue protein. These are reactive hydroxy aldehydes which can bind covalently to nucleophilic groups in biomacromolecules.e.180).172.g. and advanced glycation endproducts (AGEs) [125]. 3-O.174.168.50 – 4. the protein adducts (and to a minor extent the acyl glucuronides in their acyclic form 4.176 [123] [124].178 and 4.. 4. whose significance is now recognized due to their potential toxification [119]. 4. particularly internal migration of the acyl moiety to the 2-O. 4..171.173.170. The enaminol 4. e. respectively. 4.169. Intermolecular reactions with nucleophilic compounds include hydrolysis [107] [108]. respectively. In the resulting positional isomers 4. 4. 4. reactive carbonyl species (RCSs. These metabolites are quite reactive due to the combination of an ester and an acetal function. 4.CHEMISTRY & BIODIVERSITY – Vol. Covalent binding of reducing sugars to proteins (i. and direct transacylation of proteins [121]. . These reactions are in competition with intramolecular nucleophilic rearrangements.52.172) can undergo autoxidation catalyzed by transition metal ions. and 4. and 4.179 tautomers appear to be intermediates. transacylation with glutathione (Chapt. The resulting imine 4. and in case of abuse of acyl glucuronide-forming drugs. 5 (2008) 2221 Figs. Indeed.7) [120]. the 1-hydroxy hemiacetal is free and able to undergo the well-known reversible ring opening of reducing sugars. An important reaction catalyzed by UGTs is the formation of acyl glucuronides.175 (a Schiff base) isomerizes to a more stable form known as an Amadori compound. and 4-O positions [122]. These processes of glycation and autoxidation are known as glycoxidation and may in some cases lead to a variety of pathologies [126] in a few sensitive individuals. leading to potentially immunogenic and antigenic proteins. yielding the acyclic isomers 4.177 and enediol 4. while the products of autoxidation are reactive oxygen species (ROSs). glycation) is a phenomenon of toxicological concern.

4. 5 (2008) Fig.51. . 4. Fig.52.2222 CHEMISTRY & BIODIVERSITY – Vol.

atorvastatin. There is a nonenzymatic lactone/hydroxy acid equilibrium which is comparatively fast under gastric conditions of acidity (t1/2 of ca. among others). Of relevance here is the fact that glucuronidation of the COO group leads to an acyl glucuronide. cholesterol-lowering drugs that act by inhibition of 3-hydroxy-3methylglutaryl coenzyme A (HMG-CoA) reductase.182.183.53. A specific reaction of intramolecular nucleophilic elimination is shown by a number of statins. Enzymatic lactone-ring opening can also occur. These acyl glucuronides were characterized both as metabolites and as intermediates. . namely lovastatin and simvastatin (4. 1 h with an equilibrium constant close to one) but much slower at neutral pH [127]. and atorvastatin. prodrug form. and pravastatin) [107]. 5 (2008) 2223 Fig. i. The metabolism of statins is a complex one. The COO group in the hydroxy acid form of statins is an active target of phase-II metabolism involving conjugates as intermediates and/or metabolites. Other examples of acyl glucuronides undergoing cyclization – elimination begin to be reported [129]. 4. fluvastatin.181).e. 4. Two of the marketed statins are used in the lactone.g. while most others are used as the active hydroxy acids (e. which was detected in the in vitro metabolism of simvastatin acid (4.. in particular.CHEMISTRY & BIODIVERSITY – Vol.. by serum paraoxonase [3] [4]. since they spontaneously undergo cyclization – elimination to form the d-lactone [128].

Fortunately. We note here that the formation of albumin adducts does not prevent bilirubin glucuronidation from being a reaction of detoxification which evolved to protect animals from the intrinsic toxicity of this breakdown product. and numerous drugs metabolized to acyl glucuronides have been cleared for safety. Their incubation with human serum albumin under physiological conditions of pH and temperature resulted in a marked formation of acyl glucuronide – albumin adducts. this is not the case. predominantly to the free amino group of Lys190.186. Covalent binding occurred by the Schiff-base mechanism. causing jaundice and neurological disorders.2224 CHEMISTRY & BIODIVERSITY – Vol.52 might leave the reader with the conclusion that acyl glucuronides are consistently dangerous metabolites. 4. . and indeed the three possible acyl glucuronides (namely the two monoglucuronide isomers 4. This breakdown product of hemoglobin is markedly toxic beyond the physiological threshold. followed by biliary excretion. Figs. The endogenous UGT substrate bilirubin (4. with the usual proviso that dosage guidelines must be respected and exaggerated use avoided. In addition. and the diglucuronide 4. 5 (2008) Fig. 4.184) is of high interest in this context. a minute proportion of patients may be sensitive to the immunogenic and antigenic potential of proteins adducts [130]. Its route of detoxification and elimination is via glucuroconjugation.187) have been found in rat bile [131]. Bilirubin contains two COOH groups.54.185 and 4.50 – 4.

188) forms a comparatively stable glucuronide and few adducts. 2B15. As for the anti-allergic drug tranilast (4. the diuretic furosemide (4. the uricosuric agent probenecid (4. twelve human expressed UGTs were examined (eight in subfamily 1A and four in subfamily 2B). The latter reaction is catalyzed mainly by UGT1A1.CHEMISTRY & BIODIVERSITY – Vol. leading in some individuals to hyperbilirubinemia due to competition with bilirubin (4. protein adducts were found in the serum as well as in tissues such as the liver. Thus.189) forms a labile glucuronide and is markedly reactive toward plasma proteins [133].190) forms a comparatively stable acyl glucuronide [121a] [134]. the acyl glucuronide is reactive and undergoes in part hydrolysis or isomerization. In the case of salicylic acid (4.191) is interesting due to the presence of two target sites for UGTs. 5 (2008) 2225 Fig. While the phenolic glucuronide is stable and readily excreted. The NSAID diflunisal (4. Electronic and steric factors are postulated to determine the relative reactivities of these glucuronides. followed by covalent binding. In rats dosed with the drug. A significant finding connected with potential toxicity of acyl glucuronides is a linear relationship between their rate of degradation (hydrolysis plus acyl migration) and their propensity to bind covalently to serum albumin [123c].55.184) glucuronidation.193). were found to be active toward this substrate [136]. its main metabolic pathways in humans are O-demethylation and glucuronidation [137]. In contrast. Our last two examples illustrate the fact that many UGTs are able to form acyl glucuronides. . Our overview of acyl substrates of UGTs continues with benzoic acids [132]. In contrast. except 1A4. kidneys. and lungs [124a] [135].192). The ratio of phenolic over acyl glucuronide formed varied markedly among these enzymes. all of which. and 2B17. 4. the nonsteroidal antiinflammatory drug (NSAID) mefenamic acid (4.

195) are two arylacetate NSAIDs known for their relatively high reactivity and covalent binding to proteins [122] [123c]. Human UGT1A9 was the major enzyme involved in (S)-etodolac glucuronidation.199 formed relatively stable acyl glucuronides. covalent binding occurred by both transacylation and glycation. gemfibrozil-glucuronide was rather unstable and reactive.199) [141] and hypolipidemic agent gemfibrozil (4.196) is the active enantiomer of etodolac. whereas no substrate enantioselectivity was seen with the human 1A1 orthologue [140a].56. While 4. At pH 7.198) is another member of the profen family. This illustrates the difficulty of making qualitative reactivity previsions based on chemical structure. Its glucuronidation by rat UGT1A1 was markedly enantioselective for the (R)-form.197) were twofold faster than those of its epimer formed from (S)-ketoprofen [122] [138]. Naproxen (used as the (S)-enantiomer 4.200) [124b].to 2b-acyl migration of the glucuronide of (R)-ketoprofen (4. both of which contain hindered COOH groups. 5 (2008) Fig.194) and tolmetin (4. (S)-etodolac (4. With the exception of 2B7. with low contributions from 1A10 and 2B7. 4. Looking at chiral drugs [138]. Our last examples are the anti-HIV candidate 3-O-(3. naproxen glucuronide hydrolyzed and isomerized at similar rates [121] [140].2226 CHEMISTRY & BIODIVERSITY – Vol.4 and 378. the reaction in human liver microsomes being ca. its acyl glucuronide is of relatively low reactivity. In contrast to the previous examples. individual UGTs showed very low activity toward (R)-etodolac. The medicinal UGT substrates presented here were selected for the additional information they offer. Interestingly.3-dimethylsuccinyl)betulinic acid (4. Ketoprofen is a NSAID representative of the chiral 2arylpropanoates known as profens. the overall rate of degradation and the 1b. . fourfold faster for the active (S)-etodolac than for its enantiomer [139]. Also. Zomepirac (4. This drug is extensively conjugated in humans to a rather reactive acyl glucuronide.

Its major metabolism in humans was the N-glucuronide. 4.204) was extensively metabolized in humans by oxidation and further conjugations.57. a potent and specific maxi-K channel opener [144]. an important example is the antiepileptic drug carbamazepine (4. which proved stable in buffers and in the presence of glucuronidases. Second in importance after O-glucuronidations are the N-glucuronidations. due to the variety of target groups and substrates. . GMS-204352). as seen with sulfadimethoxine (4. Among carboxamides and sulfonamides. excepting their easier hydrolysis in the acidic pH range. The case of antibacterial sulfonamides is of toxicological significance.203. The metabolism of these drugs occurs mainly by conjugation and involves a competition between N-acetylation of the primary N(4)H2 group (see Chapt. kidney. Examples of primary and secondary sulfonamides are provided by valdecoxib and sulfonamides.202 [143]. respectively. 5 (2008) 2227 Fig. In a number of first-generation sulfonamides.205) whose N(1)-glucuronide accounted for 40 – 45% of a dose in humans [142] [146]. In addition. Lactam glucuronidation is illustrated with MaxiPost (4. In contrast. a major metabolite was its N-glucuronide resulting from direct glucuronidation at the sulfonamido group [145]. The reaction is catalyzed by UGT2B7 in microsomes from human liver. 4. Valdecoxib (4. N(1)-glucuronidation produces a soluble and nontoxic metabolite. only 2B7 catalyzed its formation.5) and N-glucuronidation at the secondary N(1)HSO2 group. and intestine.201) whose primary carboxamido group is conjugated to form the N-glucuronide 4. Also noteworthy are the species differences in the glucuronidation of tertiary amines to form quaternary Nglucuronides [142].CHEMISTRY & BIODIVERSITY – Vol. N-acetylation produced a poorly soluble metabolite which may crystallize in the kidneys and was nephrotoxic. One characteristic of many Nglucuronides compared to O-glucuronides is their greater chemical and enzymatic stability.

2228 CHEMISTRY & BIODIVERSITY – Vol. resp. 4.and N. some industrial aromatic amines known to undergo toxification to mutagenic and carcinogenic metabolites [142] [147]. witness the example of benzidine (4. N.207 and 4. by prostaglandin G/H synthase present at relatively high levels in bladder epithelium [2] [4] [149] [150]. be involved in such toxification.206) shown here [148]. 4. but their lability under even the slightly acidic conditions prevailing in the urinary tract results in their hydrolysis to benzidine (4. where a) N.58. This xenobiotic can cause bladder cancer in humans. The hepatic conjugation of benzidine by N-acetylation (see Chapt.208) is a minor metabolite due to extensive deacetylation by hydrolases [3] [4] [107].N’-diacetylbenzidine is a major metabolite which can be oxidized in the liver to reactive intermediates.207). In humans.N’-diacetylbenzidine (4. in particular. in some cases. Interestingly. Among primary and secondary amines that are substrates of UGTs.209) and mainly of N-acetylbenzidine N’-glucuronide (4.).N’diacetylbenzidine (4. . 5 (2008) Fig.206) and N-acetylbenzidine (4.210). which will then be able to undergo oxidative toxification. their N-glucuronidation may. whereas N-glucuronidation is an effective reaction yielding appreciable levels of benzidine Nglucuronide (4.5) leads to N-acetyl. in particular. and b) N-glucuronidation is minor. The metabolic situation is different in rats. we find drugs as well as a number of nonmedicinal xenobiotics.208. These two N-glucuronides are easily excreted via the kidneys. an organ-selective toxicity to be contrasted with the liver cancer selectivity prevailing in rats.

The other examples in the Figure are of medicinal interest.CHEMISTRY & BIODIVERSITY – Vol.217 [153]. it is Nglucuronidated in humans and rats. Norverapamil (4. Like a number of similar N-dealkylated metabolites of medicinal tertiary amines [147].59.211).215) is particularly interesting. since the compound contains both a primary and a secondary amino group. Like other N-glucuronides.214) is the Ndemethylated metabolite of the cardiovascular drug verapamil [152]. The upper part of this Figure completes the previous Figure by drawing the readers attention to the role played by the N-hydroxy metabolites of carcinogenic aromatic amines.213 is excreted by the kidney and undergoes acid hydrolysis in urine.212) [2] [4] [149] [150] is also a substrate of UGTs. in contrast to N-glucuronidation which produced the two regioisomeric glucuronides 4.216 and 4. Its oxidative metabolism was minimal in human liver microsomes and liver slices. its metabolite N-hydroxynaphthalen-1-amine (4. the resulting conjugate 4. The case of the antiepileptic agent D-23129 (4. . followed by toxification to the ultimate electrophilic carcinogen [2] [4] [151]. 4. Indeed and as illustrated with naphthalen-1-amine (4. Its glucuronidation proceeds mainly if not exclusively by reaction at the Natom rather than at the O-atom [142]. 5 (2008) 2229 Fig.

This compound and its active 4-OH metabolite (not shown) both form an N þ -glucuronide (4. this compound yields two direct N-glucuronides in humans. namely the quaternary N-glucuronide (4. and both were formed by incubation with human liver microsomes. In contrast.218).221 was hydrolyzed by glucuronidases and resistant to acid hydrolysis.222 was resistant toward enzymatic hydrolysis but was labile under acidic conditions. all tested UGTs. the reaction in human liver microsomes was catalyzed exclusively by UGT1A4 among all UGTs tested [155]. For both substrates. showing that even a modest structural change may alter the substrate specificity of transferases. 5 (2008) Fig.60. a number of tertiary arylalkylamines can be glucuronidated to form N-quaternary glucuronides (also known as N þ -glucuronides). see Fig. This is illustrated here with the nonsteroidal antiestrogen tamoxifen (4.36) result in such conjugates being zwitterions. The second important group of arylalkylamines forming N þ -glucuronides are N-methylpiperidines and -piperazines. .221) and the N(5)-glucuronide (4.220). catalyzed the O-glucuronidation of 4-hydroxytamoxifen. 4.222) [156]. 4. As shown in Fig. whereas 4.2230 CHEMISTRY & BIODIVERSITY – Vol. Interestingly. This permanent positive charge together with the high acidity of the carboxylate group (pKa ~ 3. 4.37. except 1A3 and 1A4. Many substrates of N þ -glucuronidation are medicinal N. Both were excreted by patients administered the drug.N-dimethylated arylalkylamines such as antihistamines and neuroleptics [142] [147] [154]. 4.219 in the case of tamoxifen). The stability of the two N-glucuronides differed markedly. as exemplified here with the antipsychotic drug clozapine (4.



Fig. 4.61. Another important group of substrates of N þ -glucuronidation are pyridines, i.e., compounds containing a pyridine ring. A typical substrate is (S)-nicotine (4.41) whose N-methylation was discussed in Fig. 4.15. Indeed, the natural (S)-nicotine (and its lactam metabolite (S)-cotinine resulting from 5’-oxidation; not shown) are N þ glucuronidated by human UGT1A4 and to some extent also by UGT1A9 [32] [157]. Nicotine N þ -glucuronide (4.223) and cotinine N þ -glucuronide are found in the urine of smokers, with large quantitative differences depending on the activity of the many enzymes involved in the metabolism of nicotine. The second example in the Figure is that of indinavir (4.224), a well-known HIV protease inhibitor. This drug features a pyridine ring which undergoes N-oxygenation and N þ -glucuronidation [158]; both are major routes of metabolism in human liver slices and in vivo. The N þ -glucuronide showed a marked species dependence in that it was formed in monkeys but not in dogs or rats. Our last example here is that of a xenobiotic, namely the tobacco smoke component 4-(methylnitrosamino)-1-(pyridin-3-yl)butan-1-one (4.225; NNK). In most tissues, this compound undergoes carbonyl reduction to 4-(methylnitrosamino)-1(pyridin-3-yl)butan-1-ol (4.226; NNAL). Both compounds are potent lung carcinogens following the oxidative toxification of their nitrosamino group. Alternative metabolic routes may thus lead to detoxification, as is the case with the glucuronidation of NNAL. Interestingly, both N þ - and O-glucuronidation occur in humans, with the two glucuronides, 4.227 and 4.228, respectively, each accounting for ca. 1/4 to 1/3 of the total NNAL excreted in the urine of smokers [159]. N þ -Glucuronidation is catalyzed in humans by UGT1A4; rats form the O-glucuronide only, a reaction catalyzed by UGT2B1.



Fig. 4.62. Besides the pyridine derivatives, a marked number of aromatic diaza- and polyazaheterocyclic compounds are known to undergo N-glucuronidation. Their interest lies in the fact that both N- and N þ -glucuronides may be formed when tautomerism is possible. Tautomerism is structurally excluded in the imidazole antifungal drug tioconazole (4.229), and indeed only the quaternary N-glucuronide (4.230) is formed, being a major urinary metabolite in humans [160]. The same reaction has been seen in a variety of N-aryl- or N-arylalkyl-substituted imidazoles, with UGT1A4 catalyzing their glucuronidation [161]. Polyazaheterocycles having an endocyclic NH (i.e., unsubstituted) moiety yield tertiary N-glucuronides, as illustrated with the methyl-1,1’-biphenyl derivatives 4.231, 4.233, and 4.236 used as model compounds [162]. Compounds 4.231, 4.233, and 4.236 contain an 1H-imidazole, a 1,2,3triazole, and a tetrazole ring, respectively. The tertiary N-glucuronides 4.232, 4.234, 4.235, and 4.237 were formed in human, monkey, dog, and rat liver microsomes at an Natom distant from the substituted C-atom, presumably due to steric hindrance. The 1,2,3-triazole ring (in 4.233) and the tetrazole ring (in 4.236) have two such distal Natoms; they are distinct in 4.233 and identical in 4.236, explaining why the former yields two N-glucuronides (i.e., 4.234 and 4.235), while the latter yields one (i.e., 4.237). Note that the tetrazole ring is of particular interest in medicinal chemistry, being an acidic group (pKa ~ 5) used as isostere of a carboxylic group in, e.g., angiotensin II receptor antagonists (sartans). The N-glucuronide 4.232 was resistant to enzymatic and chemical hydrolysis, whereas 4.234, 4.235, and 4.237 were labile. It can be hypothesized that Nglucuronide lability increases with the acidity of the target N-atom in the substrate.



Fig. 4.63. Here, we look in more detail into the problem of tautomerism. JNJ-10198409 (4.238) is a novel candidate under development with promises as an antitumor agent acting by a combination of anti-angiogenetic and antiproliferative properties [163]. As shown, the compound exists as two major tautomers at the pyrazole ring, namely the 1H- and the 2H-tautomers. The exocyclic N-atom may also be involved in a minor tautomeric equilibrium, but this is not considered here. Both tautomers at the pyrazole ring appear to be substrates of UGTs, as deduced from the formation of the tertiary N(1)-glucuronide, 4.239, in human, monkey and rat liver microsomes, and of the tertiary N(2)-glucuronide, 4.240, in rat liver microsomes (but not in human and monkey liver microsomes). Needless to say, these two N-glucuronides are different chemical entities that do not interconvert. A third glucuronide was formed in human, monkey, and rat liver microsomes, namely the N-glucuronide 4.241 which, as shown, has its two pyrazole N-atoms free to undergo tautomerism. The three N-glucuronides differed also in their chemical stability, since 4.240 and 4.241 were sensitive to enzymatic (bglucuronidase) and chemical hydrolysis, whereas 4.239 was stable. Perhaps differences in basicity can explain this contrast.

but such information might be informative in terms of catalytic mechanism and excretion. .243.242) in humans. We are not aware of any experimental evidence revealing the basicity of the N(2)-glucuronide or supporting the presence of its deprotonated form.242 reveals three possible amino > imino tautomeric equilibria. A further factor with 4. and the unchanged drug plus unidentified metabolites. 4. A case simultaneously comparable and different from the previous one is found with the anticonvulsant drug lamotrigine (4. 4. 4. 80% of a dose as the quaternary N(2)glucuronide. who excrete in their urine ca. yet their possible role during the catalytic phase cannot be discounted.243.244. 5 (2008) Fig. None of the resulting imino tautomers appears favored over the aromatic diaminotriazine structure. This pathway is the major route of metabolism of lamotrigine (4. 4. The remainder of the dose is excreted as the secondary N(5)glucuronide. two of which are targets for Nglucuronidation. Inspection of the structure of 4. to deprotonate to a tertiary N-glucuronide as shown.64. the quaternary N(2)-methyl derivative. one of which is shown.242) [164]. This compound contains five N-atoms arranged in a diaminotriazine system. the N(2)-oxide.2234 CHEMISTRY & BIODIVERSITY – Vol.242 is the possibility for its quaternary N(2)glucuronide.

251 in which the C(4)-atom is linked to b-d-glucuronic acid via a CÀC bond [167]. which features an acidic C(4)ÀH group (pKa ~ 2. An early example of the Sglucuronidation of a xenobiotic is that of the hepatotrophic agent malotilate (4.246 which is then Sglucuronidated.5-dione ring. A recent example is that of the nonsteroidal progesterone receptor agonist tanaproget (4.245).. The same reaction was seen in analogues (e.5) in the pyrazolidine-3. Its S-glucuronide 4. The formation of this metabolite implies a tautomeric equilibrium between the oxazine-2-thione moiety and its iminothiol species.250 is a major metabolic reaction in humans.65. in line with the occurrence of S-glycosides as natural products. This compound is first transformed to the dithiol metabolite 4. . 5 (2008) 2235 Fig. 4.249 has been characterized unambiguously and was a major metabolite ( > 10%) in human and rat liver microsomes [166].g.247 was excreted as a major biliary metabolite in rats [165]. Xenobiotic C-glucuronidation is an even rarer reaction than S-glucuronidation and involves very few acidic CÀH moieties.CHEMISTRY & BIODIVERSITY – Vol. phenylbutazone) and at an activated acetylenic group. yielding conjugate 4. Some examples of S-atom glucuronidation are reported in the literature.248). The resulting S-glucuronide 4. The glucuronidation of 4. The best known example is that of the uricosuric drug sulfinpyrazone (4.250).

coupled with the multidisciplinary science known as metabonomics.252).254 was the primary metabolite in the plasma of patients treated with the prodrug.256.255 were formed in microsomes from various human tissues.256 were also characterized. glycosylations may involve enzymes others than UGTs. Mycophenolic acid Oglucuronidation was catalyzed by all recombinant UGTs tested.2236 CHEMISTRY & BIODIVERSITY – Vol. Its phenol O-glucuronide 4. as suggested by the formation of xenobiotic a-d-glucosides by rat liver a-glucosidases [169]. 5 (2008) Fig.253) offers a nice example of glucosidation reactions [172].255 and the phenol O-glucoside 4. This antineoplastic and immunosuppressant agent is the active metabolite of the prodrug mycophenolate mofetil (4. since several among these enzymes are able to use UDP-glucose and transfer d-glucose to the substrate.254 and 4. human kidney microsomes produced the acyl glucoside 4. namely its phenol and COOH groups. such as lactose conjugates of antibacterial sulfonamides in the milk of lactating dairy cows [170]. and the O-glucuronides 4. Fast development in analytical and structural techniques. Mycophenolic acid (4.66. In addition. There are also reports of UDP-galacturonic acid being a cofactor of UGTs [168]. . A few anecdotical reactions have also been reported. The catalytic spectrum of UDP-glucuronosyltransferases is not limited to UDP-glucuronic acid as cofactor.257. The acyl glucuronide 4. The same was true for the phenol O-glucoside 4. thereby forming b-d-glucosides [95]. allow ever smaller traces of metabolites to be characterized and is expected to reveal yet other unexpected conjugates [171]. 4.253) contains two target sites for UGTs. Furthermore. Mycophenolic acid (4.

258. 4’’-hydroxy-CBD O-glucoside. hence the interest in their biotransformation.g. Dogs administered cannabidiol (4. yielding three acyl glucosides which were characterized in the animals urine. Plasticizers in the chemical class of phthalates are suspected of being endocrine disruptors. .67. and 6-oxo-CBD [173]. 5’’-hydroxy-CBD..260) [174]. 5 (2008) 2237 Fig. Two examples of xenobiotic O-glucosidation are presented here. The carboxy group so liberated became available for conjugation. 4. Our second example is that of bis(2-ethylhexyl) phthalate (4.261.CHEMISTRY & BIODIVERSITY – Vol. notably 4’’-hydroxy-CBD.259) and excreted as major urinary metabolites. When administered to mice.260) was oxidized on one of its 2-ethylhexyl side chains to a secondary alcohol. while the other side chain was cleaved by hydrolysis. bis(2-ethylhexyl) phthalate (4. One of these three acyl glucosides was the conjugate 4. 4. a ketone and a carboxylic acid. These metabolites were the O-glucosidated (e. CBD) produced a range of oxidized metabolites.

human UGTs of the 2B subfamily catalyzed only the former reaction. phenobarbital (4. a new aldose reductase inhibitor [179].264). Thus.263 as a minor urinary metabolite in humans. where no N-glucuronidation of barbiturates occurs.262) is conjugated to its N-glucoside 4. 5 (2008) Fig.2238 CHEMISTRY & BIODIVERSITY – Vol. C(5) in N-unsubstituted barbiturates is a prochirality center. Our second example is ranirestat (4. with a marked preference for acidic NÀH groups [175]. In the mouse. only the mouse showed a capacity to N-glucuronidate barbiturates comparable to that of humans. and product stereoselectivity in Nglucosidation was the opposite of that in humans. 4. and indeed its sugar conjugates were characterized as the ring-opened metabolites 4. barbiturate N-glucuronides and N-glucosides will exist as two diastereoisomers [178]. which were the subject of a number of informative studies published several years ago [176] [177]. the succinimide ring in ranirestat (4. Reactions of N-glucosidation seem to be somewhat better documented than reactions of O-glucosidation. Product stereoselectivity is documented in this reaction of conjugation. Note that. Among mammals. as illustrated here. However. . Typical substrates are barbiturates. to be transformed into a stereogenic center by a reaction of N-glucuronidation or N-glucosidation. and it undergoes both N-glucosidation and Nglucuronidation in humans. This compound has a succinimide ring analogous to the barbiturate ring. like the barbiturate ring. with (5S)-phenobarbital N-glucoside being the preferred product of human UGTs (probably UGTs in the 2B subfamily).264) is labile to chemical hydrolysis followed by decarboxylation. Given that sugars are chiral. phenobarbital N-glucuronide was excreted in several-fold higher amount than phenobarbital N-glucoside. Interestingly.265.68.

.272 were also seen.CHEMISTRY & BIODIVERSITY – Vol. One example is 5-aminosalicylic acid (4. N-Glucuronidation (but no N-glucosidation) was seen in humans and monkeys.5). and its formation was partly or totally nonenzymatic. Both the N-glucuronides. with a consistent preference for conjugation at the N(1)-atom. This conjugate was unstable in solution. 4. and the N-glucosides. Little N-glucuronidation and Nglucosidation occurred in rats. 5 (2008) 2239 Fig. were formed.269 and 4.267 and 4. Its major metabolite in humans is the N-acetyl conjugate (see Chapt. but traces of the N-glucoside 4.268.69. 4.and 2H-tautomers as shown. there are very few reported cases of weakly basic amino groups being N-glucosidated.266) illustrates again the interplay of tautomerism and N-glycosylation [180]. but large species differences were seen. while N-glucosidation was predominant in dogs (see Part 6). a drug used to treat inflammatory bowel disease [181]. and both NÀH groups are target of UGTs. 4. The case of the new xanthine oxidoreductase inhibitor FYX-051 (4. Besides weakly acidic NÀH groups. The compounds exists as the 1H.270.271). 4.

. and yet major species differences are apparent (see Part 6). and N-glucuronides.70. This neat classification does not correspond to well-defined families or subfamilies of UDP-glucuronosyltransferases. differ greatly in their stability. As a rule. the really significant reactions in this Chapter are the glucuronidations of hydroxy. and amino groups. To conclude this important Chapter. acyl glucuronides. From a pharmacological and pharmacokinetic viewpoint. 5 (2008) Fig. their permanent positive charge renders them zwitterionic and may be hypothesized to affect their recognition by anion transporters. carboxy. but a number of exceptions have been mentioned in this Chapter. they are usually stable toward b-glucuronidases and labile under acidic conditions. As for the N-glucuronides.2240 CHEMISTRY & BIODIVERSITY – Vol. keeping in mind that the statements made here are not rules but mere generalizations having numerous exceptions. The respective products of these reactions. O-glucuronides are consistently good substrates of b-glucuronidases and may undergo enterohepatic cycling when of sufficient molecular weight to be excreted in the bile. namely O-glucuronides. A subclass of N-glucuronides are the quaternary N-glucuronides. Thus. acyl glucuronides are reactive molecules undergoing rearrangement and forming adducts with blood and tissue proteins. 4. we summarize some of the properties of glucuronidation and glucuronides.

N-Acetylations involve mainly primary aromatic amines. 4. three reactions of acetylation are known. As shown here. However. Coenzyme A (4. As far as xenobiotic metabolism is concerned. . and hydrazides. but remain significant in a toxicological perspective [5 – 7] [182].O-acetylations. 4. The former two types of reactions use acetyl-coenzyme A (abbreviated acetyl-CoA or AcCoA. in 4. The catalytically essential group in coenzyme A is the thiol.71. which forms a reactive thioester group with acyl moieties such as acetyl (i. 4. 5 (2008) 2241 Fig.274) is an essential cofactor in the metabolism of lipids. weakly basic or non-basic NH2 groups.6 where it will be shown to play a critical role in significant yet lesser known reactions of conjugation. hydrazines. Compared to sulfonations and glucuronidations. and N. The xenobiotic substrates of O-acetylation are aromatic hydroxylamines of toxicological significance. namely N-acetylations. in other words.CHEMISTRY & BIODIVERSITY – Vol. being enzymecatalyzed reactions of trans-acetylation whose acetyl donor is a xenobiotic arylacetamide. In this Chapter.O-acetylations are not shown here and will be discussed separately. Reactions of N. some basic arylalkylamines are also N-acetylated. O-acetylations.273).e..273) as acetyl donor and are shown here. we will encounter it again in Chapt. we focus on reactions of acetylation. coenzyme A results from the conjugation of adenosine 3’-phosphate 5’-diphosphate with (R)pantothenic acid (in the larger oval) and cysteamine (in the smaller oval). they are modest in terms of the number and variety of substrates.

5 (2008) Fig.72. and its preferred substrates are arylethylamines. whose products show different yet partly overlapping substrate specificities. as we shall see. and Nat3). A number of xenobiotic basic amines have also been reported. Both the NAT1 and NAT2 enzymes are polymorphic. Nat2. Also noteworthy is the fact that the mouse has three Nat genes and three functional enzymes (Nat1. Two genes exist in humans. 4. Several of these substrates are physiological compounds such as neurotransmitters. the latter distinctly more than the former. Some large species differences have been reported in substrate specificities. The acetylation of xenobiotics is mainly a reaction of N-acetylation. hence the name serotonin Nacetyltransferase. which targets primary aromatic amines and hydrazines [8] [10] [182 – 185]. the product of the AANAT gene. with the mouse Nat1 being functionally analogous to the human NAT2 [186]. Its distribution is markedly restricted compared to NAT. The major enzyme here is arylamine N-acetyltransferase (NAT). . The second enzyme in this Figure is arylalkylamine N-acetyltransferase.2242 CHEMISTRY & BIODIVERSITY – Vol.

73. much evidence points to arylamine Nacetyltransferases (EC 2. Indeed.3.3. no O-acetylation of xenobiotics has been documented in mammals except of N-arylhydroxylamines and N-aryl-N-hydroxyacetamides (also known as N-arylhydroxamic acids).5) as the enzymes catalyzing these reactions. The literature appears rather consistent in assigning acetyl-coenzyme A-dependent O-acetylations of N-arylhydroxylamines and N-arylhydroxylamides to mainly EC 2. However.O-transacetylations to EC 3. 5 (2008) 2243 Fig.3. Another O-acetyltransferase of physiological significance is carnitine O-acetyltransferase (EC and N-arylN-hydroxyacetamide-dependent N. A number of acetyltransferases catalyze the O-acetylation of alcoholic groups in endogenous compounds. & BIODIVERSITY – Vol. but they represent enzymatic activities rather than distinct proteins.6.2. . human gene CRAT). 4. The two enzymes presented here are the ones catalyzing the O-acetylation of such xenobiotics.1. and to the best of our knowledge.56 [8]. human gene CHAT) which synthesizes the neurotransmitter acetylcholine [8]. the best known of which is choline O-acetyltransferase (EC 2.

Unchanged drug accounted for ca. 4.2244 CHEMISTRY & BIODIVERSITY – Vol.271. Several primary arylamines of medicinal interest are known to be good substrates of NAT in humans. leaving ca.276 (ca. but did not occur in dogs. whereas N(1)glucuronidation was low (ca.275) was extensively N(4)acetylated in humans to metabolite 4. An even more complex case is seen with levosimendan (4. N-Acetylation of 4. the latter metabolite has a pharmacological activity similar to that of the parent drug. the arylamine 4.9 above) (NAT1 % NAT2) [181] [182]. This is the case of para-aminobenzoic acid (NAT1) and 5aminosalicylic acid (4. suggesting NAT1 to be the enzyme involved in the reaction. an active metabolite back-acetylated to the parent compound 4.278. see Sect.278 was indeed fast in human liver cytosol (NAT2 > NAT1) and in rats (in vivo and in vitro). Other drugs include the antileprosy agent dapsone (Fig.4. see Part 6). Two plasma metabolites have been characterized in humans.57).277) [188]. 4. namely (S)-3-[4-(acetylamino)phenoxy]-2-hydroxy-2-methyl-N-[4-nitro-3-(trifluoromethyl)phenyl]propanamide (4. 35 – 40% of a dose unaccounted for [187]. Thus. 4.281 likely produced by NAT2.277. Interestingly. 5 (2008) Fig. sulfamonomethoxine (4. see Fig.280 produced by a reductive reaction catalyzed by intestinal bacteria.279) developed for the treatment of congestive heart failure [189]. .67 in Part 2 [2]). and the N-acetylated metabolite 4. 2. This agent is deacetylated in humans to the primary arylamine 4.205.275) from sulfadimethoxine (4. 11 – 13%). Our second example deals with a nonsteroidal androgen receptor modulator bearing an NAc substituent. 13 – 14%.74. There was no difference between slow and fast acetylators (NAT2 phenotypes. and various antibacterial sulfonamides. Its low glucuronidation-to-acetylation ratio distinguishes sulfamonomethoxine (4. 35 – 38% of a dose).

2.287. 4. The latter product derives from the N-Ac intermediate.284) is another good substrate of human NAT2.284 also undergoes hydrolase-catalyzed hydrolysis to hydrazine (4.286) and its monoacetyl conjugate 4. The problem is complicated by the fact that 4. The antituberculosis drug isoniazid (4.4-a]phthalazine (4. a toxic xenobiotic used in particular as a rocket propellant. until further studies proved its structure to be 3-methyl[1. 5 (2008) 2245 Fig. Earlier studies yielded contradictory results regarding the nature of this metabolite.286).CHEMISTRY & BIODIVERSITY – Vol. Besides undergoing oxidation reaction. two of which are shown here. It appears that at least part of the hepatotoxicity and neurotoxicity which threatens slow acetylators more than fast acetylators is due to free radicals derived from hydrazine (4. the antihypertensive agent hydralazine (4.282) is extensively conjugated in humans (NAT2) and other species to its N-acetyl metabolite [190].285 shows significant quantitative difference between fast and slow acetylators [191] [192].2-diacetylhydrazine (4.283 presumably via its enol tautomer. . which is unstable and spontaneously cyclizes to 4. and one whose N-acetylation to the inactive conjugate 4. Few such compounds are used as drugs. Hydrazines and hydrazides are good and highly selective substrates of human NAT2 [182].283). In contrast.75. the subsequent formation of 1.4]triazolo[3.288) is a reaction of detoxification.

76. In Sect.117) to the sulfamate 4. and the N-Ac conjugate 4.289 and acyl glucuronide 4.118. with two minor Phase I metabolites being seen.290 were the two additional pathways in these two species.289 seemingly being produced by the intestinal microflora. with acyl glucuronidation being the major one [78].289 is an interesting metabolite. rare cases of acetylation of secondary basic amines are known. . This is presumably due to the steric and electronic shielding of CYP target groups in the molecule.2246 CHEMISTRY & BIODIVERSITY – Vol. 4. Independently of its formation in the liver or intestine. The situation in dogs was more complex. Formation of the N-acetyl metabolite 4. This antibacterial agent of the quinolone class is of further interest. this N-Ac conjugate 4. we encountered the N-sulfonation of trovafloxacin (4. 5 (2008) Fig.3. While we are not aware of secondary arylamines being acetylated.5. since its metabolism is almost exclusively by conjugation in humans and rats. with no Phase-I metabolite being reported. since it is one among a rather limited number of known examples of the N-acetylation of a basic amino group. 4.

given that dogs as a rule are poor acetylators. The N-acetylation of acidic NH2 groups is also documented in a few cases.4. 4. Another example of the N-acetylation of a primary basic amine is found in the metabolism of the b-adrenoceptor blocker propranolol (4. N-Acetylcyanamide (4.e.291) rather than the parent drug. and dogs. and was a minor metabolite in humans [194].161 were accounted for by Nacetyl-N-desisopropylpropranolol (4. A different example is provided by cyanamide (4.77. 5 (2008) 2247 Fig. In this case. 2 – 4% of a dose of 4.295) is isoelectronic with the sulfonamido group. conjugation involves a metabolite (i. Indeed. rabbits. it increases acidity and hence ionization and hydrosolubility (pKa of Nacetylzonisamide is ~ 5).292) in the urine of humans administered the drug [193]. .293). however. 4. It is interesting to note that when acetylation targets slightly acidic NH2 groups (pKa of zonisamide is ca. 8% of a dose in rats. the primary amine 4.161. There were indications that the total formation of the conjugate may have been even higher given its further oxidative metabolism. a potent inhibitor of aldehyde dehydrogenase and an alcohol deterrent used as the citrated calcium salt. where basicity is canceled. accounted for ca. This contrasts with the acetylation of basic groups. whose N-Ac conjugate. and its acidic amino group is a good target of N-acetyltransferases [195].CHEMISTRY & BIODIVERSITY – Vol.295). see also Sect. rats. Cyanamide (4. 4. This last finding was largely unexpected. This is the case of the anticonvulsant agent zonisamide (4.2).294.. Another interesting finding is the evidence that NAT1 was the enzyme involved in the reaction. 10).296) is the major metabolite of cyanamide in humans.

302. is considered in a number of cases to be a reaction of detoxification. A number of industrial arylamines are known for their carcinogenic potential. the possibility remains in other cases of N-oxidation of an arylacetamide to an N-aryl-N-hydroxyacetamide such as 4. 5 (2008) Fig.298. following metabolic toxification in which reactions of N. and 4.300.2248 CHEMISTRY & BIODIVERSITY – Vol.3) or NAT-catalyzed O-acetylation to 4.g..O-sulfate ester is markedly easier when the hydrogen sulfate anion (HOSOÀ ) rather than the sulfate anion (SO2À ) is the leaving group [198]. 4. Indeed. e.297) as a prototypal example.302 (R ¼ H) forms a labile ester able to undergo heterolytic cleavage to form a highly reactive. R ¼ Ac). Nevertheless. The reactions so far discussed are marked with red arrows to signify that they are the preferred pathways of toxification of carcinogenic arylamines.. resp. 4. 3 4 This appears linked to the bladder carcinogenicity of several such arylamines. namely N.303 (R ¼ H)..g.and O-acetylation have a role to play [183b] [196 – 198]. O-Sulfonation to 4. 4. such xenobiotic arylamines can be N-oxidized to an Narylhydroxylamine (e. adduct-forming nitrenium cation 4. direct Nacetylation of xenobiotic arylamines to form.78. 4.. and ultimately to a nitrenium cation (e.303. quantum-mechanical calculations have shown that the heterolytic cleavage of an N. Interestingly.301. One important reaction is not explicit here.g. 4.O-acetylation whose mechanism is presented in more details in the next Figure. As summarized here using fluoren-2-amine (4.301 (Chapt. The latter can also be activated to a reactive O-sulfate or O-acetate. 4. .299) by cytochromes P450 (mainly CYP1A2) or the peroxidase function of prostaglandin G/H synthase (see Part 2).

In the intermolecular reaction of N. the reactions of N.CHEMISTRY & BIODIVERSITY – Vol. thus forming an acetyl thioester (NAT-S-acetyl) and liberating the Narylhydroxylamine 4.309.307 just formed remains bound to the enzyme and captures the acetyl group to yield the Oacetyl-N-arylhydroxylamine 4. another N-arylhydroxylamine. ÀN(OH)acyl).and O-acetylations [199].O-acetylation (Box D). we begin here by summarizing in Box A the reactants and products of O-acetylation. which transfers its Ac group to a cysteinyl residue in the enzyme protein. 4. 4.304. In contrast.O-transacetylations) are catalyzed by arylamine acetyltransferases. where the Ac moiety is transferred from acetyl-CoA to a generic N-arylhydroxylamine 4. However.306.79.307 (Box B).O-acetylation differs from that in N.305 and coenzyme A.310. binds to the enzyme and is acetylated to the O-acetyl-N-arylhydroxylamine 4. the products being the generic O-acetyl-N-arylhydroxylamine 4. the Ac donor in reactions of N. . the nature of the Ac donor in reactions of N. 4. 5 (2008) 2249 Fig. N. the N-arylhydroxylamine 4.308. In the intramolecular reaction of N.O-acetylation (Box C). Two options exist following this first step. Like the O-acetylation of xenobiotic N-arylhydroxylamines (À NHOH) and N-arylhydroxylamides (arylhydroxamic acids. To help the reader.O-acetylation (Boxes B – D) is an N-aryl-Nhydroxyacetamide.Oacetylation (in fact.

. 5 (2008) Fig.80. 4. its conjugation to fatty acids such as palmitic. Mention was made of the different esterification pathways open to large lipophilic alcohols. explaining its long duration of action. 4. Its incubation in ATP. 4.4.3. This endogenous steroid is also frequently used as a drug.and CoA-fortified human lung and liver microsomes yielded a variety of fatty acyl conjugates such as the palmitic ester 4. mainly endogenous and exogenous steroids. the oleyl conjugate 4. and arachidonic acids has been demonstrated in rat liver microsomes supplemented with the corresponding acyl-CoA [202]. human gene LCAT) found in blood serum. Following its inhalation. Another enzyme catalyzing the same reaction is lecithin:cholesterol acyltransferase (EC 2.1. Such highly lipophilic metabolites (e.1) and glucuronidation (Sect. The same is true of the antiasthmatic glucocorticoid budesonide (4. These reactions are independent of coenzyme A and catalyzed by hydrolases (EC 3) such as fatty-acyl-ethyl ester synthase (FAEES). cholesterol acyltransferase (ACAT)).2.2250 CHEMISTRY & BIODIVERSITY – Vol. Our first illustration is 17b-estradiol (4.26.312) [203].313.1.3.1). oleic.18) which we encountered earlier in connection with catechol O-methylation (Sect. . In Part 3. stearic. palmitoleic. The major enzyme involved in the fatty acyl esterification of such steroids is sterol O-acyltransferase (EC 2. we illustrated the fatty acyl esterification of low-molecular-weight alcohols such as ethanol.311) are stored in tissues and act as natural slow delivery devices. budesonide is extensively conjugated and retained as fatty acyl esters in airway tissues from which it is slowly released by hydrolysis.g. a microsomal enzyme of which two genes (SOAT1 and SOAT2) exist in humans. linoleic.43. and whose acyl donor is not acyl-CoA but a phospholipid [201]. which we briefly present here [8] [200].

71). Before discussing these pathways according to the classification shown. most or all of which are catalyzed by enzymes whose physiological function is in lipid biochemistry.CHEMISTRY & BIODIVERSITY – Vol. The key compound here is coenzyme A (4.81. yielding xenobiotic acyl-CoA cofactors which are a crossroads to an unusual variety of further metabolic pathways. 5 (2008) 2251 Fig. However. many of them are synthetic (anabolic) ones. These pathways will be presented.5 and the present one are centered [205]. and the numerous metabolic pathways open to such acyl-CoA cofactors. the present Chapter has xenobiotic acids as substrates. The coupling of xenobiotic acids to coenzyme A. as schematized in this introductory Figure. on which both the previous Chapt.273 in Fig. Their conjugation to coenzyme A is catalyzed by ligases. an obvious exception being b-oxidation. 4. . 4. in turn. form a complex and insufficiently understood field of research at the interface of lipid biochemistry and xenobiotic metabolism [204]. there is a major difference between the two Chapters. Whereas the previous Chapter dealt with endogenous acids (acetic acid and fatty acids) bound to coenzyme A and transferred to a xenobiotic acceptor. we examine the ligases involved in xenobiotic acyl-CoA formation and some properties of the resulting cofactors. 4.

There is an overlap in the substrate specificity of these enzymes. The enzymatic reactions take their energy from the splitting of ATP into AMP and pyrophosphate. The coupling of carboxylic acids to coenzyme A is catalyzed by enzymes known as acid-thiol ligases (EC 6. 4. which includes one or more very long-chain acyl-CoA ligases. all the more so.1).2252 CHEMISTRY & BIODIVERSITY – Vol.82. a large variety of xenobiotic carboxylic acids are substrates of these ligases. these are the short-chain acylCoA ligase. The subcellular and tissular localizations differ among enzymes. As we shall see.2. the medium-chain acyl-CoA ligase. three of which play a leading role in forming xenobiotic acyl-CoA cofactors [204 – 207]. and the long-chain acyl-CoA ligase. but each of the resulting acylCoA conjugates is then processed by only a few among the pathways depicted in the previous Figure. . 5 (2008) Fig. since they are in fact families with various isozymes as documented by the human genes reported in the Figure. As shown. with the magnesium cation acting as a cofactor in the reaction.

acyl-CoA thioesters react as electrophiles with H2O (chemical hydrolysis) and with glutathione (see Chapt. the inhibition of fatty-acid b-oxidation is documented for some medicinal arylalkanoic acids. 5 (2008) 2253 Fig.1.CHEMISTRY & BIODIVERSITY – Vol. acyl-CoA conjugates also undergo a variety of reactions summarized in this Figure. the mechanism of this inhibition does not involve their acyl-CoA conjugates but appears to be due to a combination of coenzyme A sequestration and direct inhibition of acyl-CoA ligases [205] [215].1. some xenobiotic acyl-CoA conjugates are pharmacologically active.6. On a more positive side. There is also evidence that clofibric acid and other fibrate drugs used as hypolipidemic peroxisome proliferators act as their acyl-CoA conjugates to inhibit fatty-acid synthesis [214]. Acyl-CoA conjugates are also specific substrates of acyl-CoA thioesterases (EC 3. . The main focus of this Chapter is on the metabolic reactions typical of xenobiotic acyl-CoA cofactors. but there is proof that the acyl-CoA conjugate of the inactive (R)-enantiomers also contributes to the therapeutic effects by inhibiting COX-2 [213]. the (S)-enantiomers of the well-known non-steroidal anti-inflammatory drugs (NSAIDs) ibuprofen and ketoprofen (see Sect.2) [210]. Thus. 4.4) are the active compounds that inhibit cyclooxygenase (COX). Finally. In addition and as a result of their intrinsic reactivity [208]. particularly profens. 4. A worrying property is the structure-dependent capacity of some xenobiotic acyl-CoA conjugates to react as electrophiles with proteins. a series of xenobiotic acyl-CoA conjugates revealed a correlation between their rates of hydrolysis and their rates of transacylation with glutathione [209a]. Thus. Interestingly.2. 4.7) to form glutathione conjugates as shown [209].2. thereby forming acylated proteins with immunogenic potential [211] [ 212].1 and mainly 3.83.

The most common amino acid in such reactions is glycine. and its prototypal substrate is benzoic acid (4.318. since it was first characterized in 1829 by Liebig in the urine of horses [218].192). in humans. The same is true for acetylsalicylic acid (4. Aspirin). The enzymes catalyzing the formation of N-benzoylglycine (4. In contrast to the acyl-CoA cofactors whose lability and metabolic intermediacy prevents their excretion from the body. more precisely its benzoyl-CoA cofactor [217].84. Further examples of medicinal interest include the hypolipidemic drug nicotinic acid (4. Hippuric acid (4. and GLYATL2). .315) are mainly glycine N-benzoyltransferase (EC 2.71) and glycine N-acyltransferase (EC 2.315) is thus considered by many to be the first xenobiotic metabolite ever characterized. GLYATL1.319) [222]. Note that N-benzoylglycine is better known under its trivial name hippuric acid. most of the subsequent metabolites are sufficiently stable and hydrophilic to be excreted.2254 CHEMISTRY & BIODIVERSITY – Vol. which are too lipophilic to be excreted. over 90% of a dose is hydrolyzed to salicylic acid (4. Exceptions are the lipid and sterol conjugates. most of which (depending on the dose) is eliminated as salicyluric acid (4.1.1. 4.314). human genes GLYAT. nicotinuric acid) is a major metabolic pathway [221].13.3. Numerous ringsubstituted derivatives and analogues of benzoic acid form a glycine conjugate [219].316) whose conjugation to N-nicotinoyl glycine (4.3.317. giving insight into informative structure – metabolism relationships [132] [220]. Here. we begin with amides formed from an endogenous amino acid and a xenobiotic acyl-CoA cofactor [206] [216]. 5 (2008) Fig.

6. Our first example is 4-phenylbutanoic acid (4. which produces high levels of acyl glucuronides. 4.322 (30 – 35%). In contrast. plus N-(2-phenylacetyl)glycine ( the formation of glutamine conjugates presented here favors larger substrates. There is more to the metabolism of this drug. The latter metabolite is a unique observation in humans. allowing for possible exceptions (see later). This appears indeed as a rule.320) helps nitrogen excretion in the form of its glutamine conjugate 4. This compound formed only traces of its glycine conjugate in epileptic patients.321. These metabolites are not formed in the rat. the metabolism of 4-phenylbutanoic acid (4. The previous Figure suggests that only benzoic acids and small analogues yield glycine conjugates in humans. while its glutamine conjugate 4. Indeed. Two enzymes are the main catalysts of the reaction.4) to form 2-phenylacetic acid also excreted as the glutamine conjugate 4.325 and glutamic acid conjugate 4.68) which does not act on benzoic acids.CHEMISTRY & BIODIVERSITY – Vol.324).1. an effective drug used in patients with hyperammonemia due to inborn errors in urea synthesis.85. being formed either by conjugation with glutamic acid or by deamidation of the glutamine conjugate. The Figure also shows one of the many metabolic pathways of the antiepileptic drug valproic acid (4. 4.3. 5 (2008) 2255 Fig. and glutamine N-acyltransferase (EC 2.323) not found in humans. since it undergoes C2 chain shortening by b-oxidation (see Sect.14) which shows a broad substrate specificity. namely glutamine N-phenylacetyltransferase (EC 2. .326 were produced in somewhat higher amounts [224]. a major metabolite in humans (20 – 25% of a dose) [223].

Its taurine conjugate 4. 2-(4-chloro-2-methylphenoxy)acetic acid (4. formed both the taurine conjugate 4. no taurine conjugate and only traces of the glycine conjugate were formed and excreted in rats. the conjugation of bile acid-CoA with taurine or glycine is catalyzed by the cytosolic enzyme bile acid-CoA:amino acid N-acyltransferase (EC 2. Both metabolites were of marked importance in the dog.6. There have been a number of recent reports of xenobiotic carboxylic acids yielding taurine conjugates. which excreted most of a dose unchanged.332).3.333 was a minor urinary metabolite (1 – 2% of a dose) excreted almost exclusively as the (S)-enantiomer [228]. 4.327). In contrast to 2-(4chloro-2-methylphenoxy)acetic acid (4. Low doses favored the taurine conjugate.328 and the glycine conjugate 4. In humans. . shares an important physiological function in the conjugation of biliary acids.327). together with glycine. Thus. One nice example is that of the NSAID ibuprofen (4. human gene BAAT) [8] [225]. dose-dependent amounts of the acylglucuronide and taurine conjugates were excreted in the bile.65.1. whose proportion decreased at higher doses. in contrast to the withdrawn NSAID benoxaprofen whose taurine conjugate was mainly the (R)enantiomer in rats (see also Sect. When administered to dogs. the retinoid X-receptor ligand and antitumor agent targretin (4. an agonist of the a-peroxisome proliferator-activated receptor (aPPAR) of interest as a cholesterol.86. Taurine (H2NÀCH2CH2 ÀSO3H) is another amino acid which.4). 5 (2008) Fig. There are few reports of xenobiotic taurine conjugates being formed in humans. a plant growth regulator used as herbicide.329 [226].330).and triglyceridelowering agent [227a]. 4.2256 CHEMISTRY & BIODIVERSITY – Vol. Another interesting example is that of MRL-II (4.331) did form taurine conjugates in rats from the unchanged drug and from two oxidized metabolites [227b]. In contrast.

e.e. and MOGAT3). hybrid phospholipids such as the generic 1-acyl2-palmitoyl-3-phosphocholine 4. 4. 2-acylglycerol O-acyltransferase (EC2.1. the resulting conjugates are classified as more lipophilic than the parent xenobiotic carboxylic acids (hybrid lipid. hybrid) glycerides and sterol esters formed from xenobiotic acids [204] [229] [230].336.335.3. or more hydrophilic (carnitine esters). 1-acylglycerophosphocholine O-acyltransferase 1 (EC 2.3.3. MOGAT2. 5 (2008) 2257 Fig. and ethyl esters). The formation of hybrid triglycerides and sterol esters has been reported for a number of xenobiotic carboxylic acids using.334.22. sterol O-acyltransferase (see Fig..87.CHEMISTRY & BIODIVERSITY – Vol.1.81.g. human genes MOGAT1. most notably diacylglycerol O-acyltransferase (EC 2. we summarize the major types of mixed (i.80).1.. human gene PLCAT1). In vivo evidence in animals is also available. 4. A number of enzymes are known or believed to catalyze the formation of lipid conjugates.1. .23. and acylglycerol-3-phosphate O-acyltransferases (EC 2.20.51 and -52. These conjugates include hybrid triglycerides such as the generic dipalmitoylglyceride conjugate 4. Based on their lipophilicity and resulting behavior in the body. sterol esters. and cholesteryl esters 4. human or animal hepatocytes or adipocytes [230]. human genes DGAT1 and DGAT2). Here. The esters formed by the acylation of endogenous hydroxy compounds with xenobiotic acyl-CoA cofactors are summarized in Fig. mostly in rats [231].3. human gene root AGPAT) [8]. 4. There are also examples of xenobiotic carboxylic acids being coupled to the 3-OH group of bile acids [232].

this conjugate was shown to be the causative agent of microgranulomatous changes in the liver of mice chronically fed fenvalerate (4. the cholesteryl ester was formed and accumulated in the liver. An example of cholesteryl ester formation is provided by the hypolipidemic agent 4-[10-(4-chlorophenoxy)decyloxy]benzoic acid (4.337) [234]. triacylglycerol analogues were the major products formed in cultured adipocytes.340) in rats and mice [236]. Benzoic acid (4. Accumulation in the liver and other tissues is certainly a factor of concern. witness the herbicide 4-(2.2258 CHEMISTRY & BIODIVERSITY – Vol. Remarkably. A few studies have investigated the rate of elimination of hybrid lipid conjugates from tissues and found it to be markedly slower than that of their natural counterparts [231b] [237]. .341). A marked number of xenobiotic carboxylic acids have been reported to form hybrid lipid conjugates [229 – 231].314) is unusual. while some mixed diacylglycerols and phosphatidylcholines (i. Following oral administration to rats..340). In this case. and some hypolipidemic drugs such as lifibrol (4. the antiepileptic valproic acid (4.4-DB) [230]. This pyrethroid insecticide exists as four stereoisomers. 2. 4. being neither metabolized further nor transported by lipoproteins.4dichlorophenoxy)butanoic acid (4.339) [235]. a lipophilic residue was identified in all analyzed tissues and found to be the cholesteryl ester of (2R)-2-(4-chlorophenyl)isovaleric acid (4. Some agrochemicals are also substrates. Several drugs are substrates.e. which was incorporated into phospholipids in cultured neurons [233]. Following administration of the isomeric mixture. Other drugs include NSAIDs such as ibuprofen (4. since the vast majority of known substrates are larger molecules. as illustrated by the fate of fenvalerate (4.324).332) and ketoprofen [231]. containing one fatty acid and one xenobiotic acid) were also observed. 5 (2008) Fig.88. for example.338. as illustrated here.

1.344. Etretinate is an ethyl ester prodrug whose hydrolase-catalyzed hydrolysis yields the active carboxylic acid known as acitretin (4. we note that another interesting metabolite of acitretin is (Z)-acitretin (4. Interestingly. 4. This thioester is then transesterified to etretinate by an insufficiently characterized ethanol acyltransferase which can also use other short-chain alkanols [239].2. the highly lipophilic etretinate being stored in adipose tissues and serving as a reservoir for the active acitretin (4. 4. a reaction catalyzed by human liver microsomal long-chain fatty acid-CoA ligase (see Fig. In patients under long-term therapy. plasma etretinate levels remain detectable for over two years following discontinuation of treatment. the CoA-thioester 4. There are only few examples of the coenzyme A-dependent esterification of xenobiotic carboxylic acids with short-chain alkanols. In fact and to the best of our knowledge.2.345 (resulting from an initial C1 shortening in a 3-methyl-branched carboxylic acid ¼ a-oxidation) and 4.343). a highly teratogenic retinoid used cautiously for the treatment of severe psoriasis resistant to other forms of therapy.7. discussed below).1. The explanation was found with the discovery that part of the metabolically formed 4. the only example of medicinal interest is that of etretinate (4. As an aside.343 forms the acitretinoyl-CoA thioester conjugate 4. . 5 (2008) 2259 Fig.346 (resulting from subsequent C2 shortening ¼ b-oxidation.342).344 is also the intermediate in the formation of side-chain-shortened metabolites such as 4.82).343) [238].343).CHEMISTRY & BIODIVERSITY – Vol. The resulting futile metabolic cycle is in fact far from pharmacologically futile.347) whose reversible formation is presumably catalyzed by the retinoid cis-trans isomerases EC 5.89. a pharmacokinetic result incompatible with effective hydrolysis and further degradation and elimination of acitretin (4.3 and 5.

While Gly. human genes CPT1A. .1. Valproic acid (4. carnitine O-octanoyltransferase (EC 2. l-Carnitine ((R)-(3-carboxy-2-hydroxypropyl)(trimethyl)ammonium) is a d-amino acid whose essential role is to allow membrane translocation of fatty acids following their conjugation to acyl-carnitines.349 > carnitine conjugate 4.21. also forms amino acid conjugates. as seen in the urine of epileptic patients treated with the drug [241].324).7. and carnitine O-palmitoyltransferase (EC 2.350 > glycine conjugate [211a]. and taurine form amide conjugates with xenobiotic acids.90. the relative quantitative importance of conjugates was acyl glucuronide > taurine conjugate 4. human gene CROT) which acts with C4 – C16 alkanoic acids. in particular carnitine O-acetyltransferase (EC 2.1. since its trimethylammonium group is not available for conjugation. The anti-inflammatory agent tolmetin (4. CPT1C. also forms valproylcarnitine (4.4. whose capacity to form hybrid glycerides was presented above.137.195). 4. and CPT2) having a broad substrate specificity in the C6 – C20 range [8] [240]. The formation of the latter is catalyzed by carnitine O-acyltransferases.3. 4.1.348) as a minor metabolite in humans. whose acylglucuronide was discussed in Chapt. Another physicochemical characteristic of carnitine conjugates is the fact that the negative charge in the parent carboxylate anion is replaced by a zwitterionic function capable of forming an internal ionic bond. In rats. for example.3. CPT1B.3. 5 (2008) Fig. Two medicinal examples of carnitine conjugates are shown here.2260 CHEMISTRY & BIODIVERSITY – Vol. human gene CRAT) which acts with C2 – C10 alkanoic acids. Glu. carnitine forms esters via its OH group.

However. the formation and excretion of pivaloylcarnitine (4.91. .353).CHEMISTRY & BIODIVERSITY – Vol. A medicinally relevant situation is that of pivampicillin (4.356).355). a known substrate of the carnitine conjugation pathway.351). Bio-hydrolysis of pivampicillin liberates the promoiety pivalic acid (4. 4.356) > cyclobutanecarboxylic acid > cyclohexanecarboxylic acid ¼ benzoic acid > pivalic acid. a miticide agent whose hydrolysis liberates cyclopropanecarboxylic acid (4. 5 (2008) 2261 Fig. The latter is a good substrate of carnitine O-acetyltransferase. This Figure exemplifies cases where the substrate of carnitine conjugation is not the xenobiotic per se but a metabolite thereof. as shown in rats receiving high doses of cycloprate [243].352). In rat hepatocytes. Humans supplemented with carnitine excreted up to 90% of the administered amount of pivalic acid as pivaloylcarnitine [242]. the substrate specificity of carnitine ester formation was cyclopropanecarboxylic acid (4. the (pivaloyloxy)methyl ester of the b-lactam antibiotic ampicillin (4.354) in humans and rats dosed with pivalic acid or pivampicillin is limited by the body reserves of carnitine. The second example is that of cycloprate (4.

35). 4.1. See also Fig. 4.1.82). .2262 CHEMISTRY & BIODIVERSITY – Vol.1. see Fig. 4.83) to 10-phenyldecanoic acid (C10-4.2. and further on all the way to 2-phenylacetic acid. In addition.358). their physiological products being acetyl-CoA and ATP. 5 (2008) Fig. Thus.3. Most available examples concern b-oxidations. The sequential b-oxidation steps involve 2.3.3-dehydrogenation catalyzed by acyl-CoA oxidase (EC 1. The resulting 10-phenyldecanoyl-CoA (C104.3. it is substrate of a subsequent cycle of b-oxidation to 8-phenyloctanoic acid. Extensive investigations with wphenylalkanoic acids 4.358 have revealed that such xenobiotic carboxylic acids are also good substrates of the b-oxidation enzymatic machinery [8] [244] [245]. hydration to the (S)-3-hydroxyacyl intermediate catalyzed by enoyl-CoA hydratase (EC 4.1. a reaction catalyzed by long-chain acyl-CoA ligase (EC in which the b-oxidation to 4-phenylbutanoic acid (4. which in eukaryotes occur in peroxisomes (for very-long-chain fatty acids) and mitochondria.2. 4.3.358-CoA).17). lengthening its chain.2.358-CoA) can be hydrolyzed by fatty acyl-CoA hydrolase (EC 3. or racemizing it.2.1. see Fig. 12phenyllauric acid forms its CoA-conjugate (C12-4. we survey reactions of xenobiotic acyl-CoA conjugates that modify the acyl moiety by oxidizing it. In this last Section of Chapt. and oxidative cleavage with loss of acetyl-CoA catalyzed by 3-oxoacyl-CoA thiolase (EC 2.16).6). dehydrogenation to the 3-oxoacyl intermediate catalyzed by 3-hydroxyacyl-CoA dehydrogenase (EC 1.320) was alluded to. 4.6.

5 (2008) 2263 Fig. it was incorporated in rat myocardium.359) [246]. In the second pathway.362. 4. this C¼C bond was reduced by an NADPH-dependent reductase to yield 4. abbreviated as [99mTc]CpTT-PA) [247]. In the first pathway. Interestingly.93. recognized as a fatty acid. The C12-shortened analogue (produced by six cycles of b-oxidation) was the major metabolite found in heart lipids and heart perfusate.CHEMISTRY & BIODIVERSITY – Vol. and metabolized as such by b-oxidation.364. indeed. A more complex case of b-oxidation is presented by the thromboxane A2 receptor antagonist known as (þ)-S-145 (4. the parent compound was substrate of a first cycle of b-oxidation to yield the C2-shortened metabolite 4. . A recent example of b-oxidation is provided by [99mTc]tricarbonyl(15-cyclopentadienyl pentadecanoic acid)technetium (4. This compound has been developed as a potential diagnostic tool in heart diseases.361.363 first necessitated reduction (or perhaps 3 ! 2 displacement) of the C¼C bond. its fatty-acyl side chain indeed underwent boxidation. two independent pathways of b-oxidation were characterized due to the presence of the (Z)-configured C¼C bond at the 5-position of the side chain. When incubated in rat hepatocytes and rat liver peroxisomes. Further b-oxidation of the latter to produce the C4-shortened metabolite 4. And.360 before b-oxidation produced the C2-shortened metabolite 4.

4.26. but it was not seen upon incubations of 4. the compound was oxidized at various positions in its alkyl side chain. namely lovastatin and simvastatin (4. In rats in vivo and in vitro. while most others are used as the active hydroxy acids.6-trimethoxyphenyl)dodecanamide (4. and indeed their hydroxy-acid form is known to yield an acyl-CoA thioester. a C5-shortened homologue was produced as a minor metabolite. The two previous Figures illustrate the b-oxidation of xenobiotic carboxylic acids. yielding ketones and the carboxylic acid metabolite 4. .4. these are not the only cases of interest here. ACAT) with potential as an inhibitor of cholesterol absorption and deposition [248]. This intriguing metabolite further illustrates the complexity of the endogenous metabolic pathways which xenobiotic carboxylic acids may enter. In addition. When incubated with liver microsomes from humans or animals. 4. However. A somewhat different picture emerges with the statins.365.2-dimethyl-N-(2. the latter metabolite underwent three cycles of b-oxidation to produce the C6shortened carboxylic acid 4. Several statins including simvastatin and lovastatin have been shown to undergo one or two cycles of b-oxidation [249]. two of which are used in their lactone. CI-976). This situation is exemplified here with 2. see Fig.366.366 or its w-OH precursor.2264 CHEMISTRY & BIODIVERSITY – Vol.92). 5 (2008) Fig.53).3.367.181. 4. However. an inversion of configuration is necessary and occurs via dehydration at C(2) and C(3). the (R)-configured 3-OH group in statins prevents b-oxidation (see Fig.94.1. as shown here. prodrug form. a highly effective inhibitor of cholesterol Oacyltransferase (EC 2. since w-oxidation of an alkyl side chain in a xenobiotic compound may also yield a metabolite able to undergo b-oxidation.

1.1. 4. This metabolite was formed in rats by the oxidative dechlorination of the dopamine b-hydroxylase inhibitor 5-(4-chlorobutyl)picolinic acid (4.373 to 4.371).95.3.CHEMISTRY & BIODIVERSITY – Vol. This reveals the remarkable fact that several enzymes in b-oxidation also catalyze the reverse reaction. Also worthy of note is the fact that all known reactions of chain elongation involve a single C2 unit. and they represented stable intermediates in a cycle of C2 addition. The four metabolites are arranged here in a biochemically logical sequence allowing comparison with b-oxidation (Fig.3.6) which initiates boxidation.1. These metabolites were also produced from exogenous (radiolabeled) benzoic acid.374 is catalyzed by EC 1.373 was most likely catalyzed by EC 2. An example is that of benzoic acid (4.3.374 to 4.16 (here known as acetyl-CoA C-acyltransferase). The last step is a hydrogenation catalyzed by the NADPH-dependent trans-2-enoyl-CoA reductase (EC 1. except for cyclopropanecarboxylic acid (4.92).373 to 4. while dehydration of is expected to be catalyzed by EC 4.372 to produce 4.17 acting as a dehydrase.376 [204] [251]. an enzyme distinct from the flavoprotein acyl-CoA oxidase (EC 1. Even more informative is the case of 5-(3-carboxypropyl)picolinic acid (4. Reduction of 4. acetyl-CoA transfer to the CoAconjugate of 4.38). Indeed.35 acting as a hydrogenase. . and it was excreted mainly as the C2-elongated metabolites 4.372).369) [250].2. Note that the elongation of 4.314) which in the horse yielded small amounts of 3-hydroxyphenylpropionic acid (4. 5 (2008) 2265 Fig.3.372 was also seen in other animal species including humans. The chain elongation by C2 units is known for a small number of substrates [204]. 4.370) and possibly also 3-oxo-3-phenylpropanoic acid (4.356) which was elongated up to C16 depending on animal species [204].

this intermediate is the substrate of a reaction under inversion of configuration catalyzed by 2-methylacyl-CoA 2-epimerase (EC 5.332).82) [206] [254].1.. a reaction catalyzed by long-chain acyl-CoA ligase (EC 6. As a result of epimerization. which are now fairly well understood. an extensively used drug [252] [253]. This reaction is one of racemization when considering only the substrate moiety. a-methylacyl-CoA racemase.4. 4. the ibuprofenoyl moiety now exists in the (R). . it is one of epimerization.2. the first step is the formation of an acyl-CoA intermediate. in contrast.e.96. not an entry point.99. Once formed. namely the inversion of sense of chirality of non-steroidal anti-inflammatory 2arylpropanoic acids (i. In other words and in the case of ibuprofen. It took many years and a number of research teams to characterize the enzymology and mechanism of the reaction.2266 CHEMISTRY & BIODIVERSITY – Vol. This reaction is enantioselective in that it shows a marked or practically exclusive preference (depending on animal species) for the inactive (À)-(R)-profens [255]. 4.1. in strictly correct terms. (R)-ibuprofen is both. but. human gene AMACT). As shown here. 4. a peroxisomal and mitochondrial enzyme catalyzing an essential step in the oxidation of cholesterol to cholic acid [256] [257].3. since the acylCoA conjugate contains several stereogenic centers (see Fig. Fig. In the metabolic scheme shown here. profens). the most investigated of which is ibuprofen (4. and acylCoA thioesterases act on both (R)-ibuprofenoyl-CoA and (S)-ibuprofenoyl-CoA to liberate the corresponding ibuprofen enantiomer. the ligase forms only or mainly the (R)-ibuprofenoyl-CoA. 5 (2008) Fig.71). An intriguing metabolic reaction has attracted much interest since the 1970s.and (S)-forms. (S)-ibuprofen is thus an end-point only.



Fig. 4.97. A few profens beside ibuprofen have been shown to undergo inversion of configuration in humans, namely ketoprofen (4.377), fenoprofen, and benoxaprofen [258] [259]. In vivo and in vitro results in a variety of animal species indicate that the extent of inversion of profen depends strongly on substrate as well as on species [252] [260]. Furthermore, inversion of configuration is not restricted to 2-methylsubstituted arylacetic acids, witness KE-748 (4.378), the active metabolite of the antirheumatic agent KE-298 [261]. Like ibuprofen, KE-748 underwent extensive (R)to-(S) inversion when administered to rats or incubated with rat hepatocytes. An apparently unconnected example is that of phytanic acid (4.379), namely (3RS,7R,11R)-3,7,11,15-tetramethylhexadecanoic acid. This branched fatty acid is present in various dietary sources, in particular in the fat of ruminant animals where it accumulates as a metabolite of phytol, itself a decomposition product of chlorophyll. In humans and animals, phytanic acid is conjugated to coenzyme A, but its 3-methyl substitution forbids subsequent b-oxidation [244]. Instead, the phytanoyl-CoA intermediate is substrate of a-oxidation to form the coenzyme A conjugate of pristanic acid, more accurately the two epimers (2S)- and (2R)-pristanoyl-CoA (4.380). bOxidation to form propanoyl-CoA is now possible, but only for (2S)-pristanoyl-CoA; 2-methylacyl-CoA 2-epimerase is necessary to catalyze the inversion of configuration of (2R)-pristanoyl-CoA and avoid accumulation of (2R)-pristanic acid [256] [257] [262]. To us, phytanic acid (4.379) thus appears as an example at the interface of xenobiotics and alimentary compounds.



Fig. 4.98. This Figure opens an important Chapter dedicated to glutathione (4.385; GSH), a tripeptide playing an essential role in the protection of organisms against a variety of environmental insults. We initiate this Chapter with the biosynthesis of glutathione (4.385) as schematized in the left part of the Figure [263] [264]. The reaction begins with the formation of g-glutamyl-cysteine (4.383) from l-cysteine (4.381) and l-glutamic acid (4.382), as catalyzed by g-glutamylcysteine synthetase (EC Glycine (4.384) is introduced in the second step catalyzed by glutathione synthetase (EC; both reactions are ATP-dependent. The all-important and limiting component of GSH is cysteine, whose SH group is responsible for its conjugating and antioxidant capacities. Indeed, glutathione is not only an important conjugating compound, it is also an essential antioxidant agent, so essential that its physiological concentrations in cells and plasma are in the 0.5 – 10 mm and micromolar range, respectively [265]. In the body, GSH exists in a redox equilibrium with an oxidized form known as glutathione disulfide (GSSG) [266]. Glutathione acts mainly as a radical scavenger as summarized in the right side of the Figure [267 – 269]. Like other endogenous thiols including serum albumin, glutathione scavenges free radicals, in particular C-centered radicals (Reaction 1) and reactive oxygen species (ROSs) such as the HO . radical, superoxide, and peroxide radicals (Reactions 2 – 5). These reactions transform GSH into the glutathionyl radical (GS . ) which is detoxified by reacting with a second GS . radical to yield GSSG (Reaction 6). The latter is recycled to GSH by glutathione disulfide reductase (EC; Reaction 7) [263] [264]. As such, GSH plays a critical role in cellular protection against oxidative stress and radiations [270].



Figs. 4.99 and 4.100. Whatever the significance of radical scavenging reactions in the cellular protection, our scope remains on xenobiotic metabolism, and specifically here on GSH-dependent conjugations. These are catalyzed by glutathione S-transferases (EC; GSTs), a large and diverse group of enzymes present in the cytoplasm, in the endoplasmic reticulum, in mitochondria, and in peroxisomes [10] [271 – 274]. Not one but two superfamilies of genes exist in animals, namely those coding for the cytoplasmic superfamily of enzymes (GSTs in the narrow sense), and those coding for the microsomal superfamily of enzymes (designated as MGSTs when distinguished from the cytoplasmic GSTs). In humans, the microsomal enzymes are the products of three MGST genes, and they function as homotrimers. The cytoplasmic (i.e., soluble) enzymes are products of the GST genes, a large number of which are known. These enzymes function as homodimers or heterodimers as shown. The main functions of these enzymes are the formation of conjugates of endogenous and exogenous compounds, and these are the relevant functions we will present here. In addition, these enzymes perform other physiological functions such as isomerization, reduction, thiolysis, and transport which fall outside our scope.

2270 CHEMISTRY & BIODIVERSITY – Vol.100. 5 (2008) Fig. 4. .

103. as shown in Fig.0 and is thus poorly ionized at physiological pH. 4. the reactivity of the substrate is increased by polarization of its electrophilic center. which in aqueous solution has a pKa of 9. Fig.101 schematizes the transition state in a reaction of glutathione addition – elimination whose substrate here is 1-chloro-2. This Panel shows the initial catalytic step in reactions of glutathione addition.101 – 4. 4. the high structural flexibility of glutathione S-transferases (GSTs) is not taken into account here [279]. Both Panels a and b are highly simplified representations (modified from [275a]) which do not incorporate further residues involved in catalysis and/or substrate binding by hydrophobic and electrostatic interactions [278].101 for 4-phenylbut-3-en-2-one (Panel a).CHEMISTRY & BIODIVERSITY – Vol. Significantly. Furthermore. a stabilization achieved in particular by an H-bond donation from a neighboring tyrosine residue. 4. 4.101 focuses on some common features in the catalytic mechanism of glutathione S-transferases [275] [276]. In turn. 4. and the thiolate anion in GS À is thus the catalytically active group in GST-bound glutathione.4dinitrobenzene.103. A main element in this mechanism is the acidity of the SH group in glutathione. This 100.102. whose main types of substrates and products are reviewed in Fig. Panel b in 1000-fold increase in acidity results from a considerable stabilization of the ionized thiol. The main types of substrates and products of these reactions are reviewed in Fig. SH acidity is markedly increased (pKa in the range 6 – 7) when GSH is bound to glutathione Stransferases [277]. 5 (2008) 2271 Figs. . This increased acidity is translated into an increased nucleophilicity toward reactive electrophiles. 4.

4. .2272 CHEMISTRY & BIODIVERSITY – Vol. 5 (2008) Fig.102.103. 4. Fig.

in vitro and in vivo. a given chemical containing a potential target group may be a good or a poor substrate of glutathione transferases. Thus. followed by the addition of an aliquot of rat liver cytosol and a further 1-min incubation [280a]. will be mediated mainly (Case 1) or solely (Case 2) by GSTs. 4-Nitrobenzyl chloride (4. 4. the first two cases involve reactions which.CHEMISTRY & BIODIVERSITY – Vol.e. An intriguing aspect of glutathione conjugation is the potential occurrence of nonenzymatic reactions [280]. 1.4dinitrobenzene (4. Case 3 involves mainly or exclusively nonenzymatic reactions. 5 (2008) 2273 Fig. In an enlightening and still current review.104.390) was almost inert..389) was a poor GST substrate but reacted spontaneously with GSH. the reactions were monitored spectrophotometrically and involved a 1-min incubation of the xenobiotic with GSH in the absence of enzymes. it may also be able or unable to react spontaneously (i. enzymatic GSH conjugation of five xenobiotics.386) and 1-chloro-2. 1-(4-Nitrophenoxy)propane 2. nonenzymatically) with glutathione in solution.3-oxide (4.388) was a fair GST substrate and reacted spontaneously with GSH. whereas Case 4 comprises chemicals resistant to GSH conjugation. The four resulting combinations are shown here. offering a cogent classification into four possible scenarios [280a].2-Dichloro-4-nitrobenzene (4.387) are seen to be good GST substrates and to react spontaneously with GSH. The remainder of the Figure compares the nonenzymatic vs. whereas (E)-4-phenylbut-3-en-2one (4. . Ketterer has highlighted the role of nonenzymatic glutathione conjugation reactions in xenobiotic metabolism.

and as such will react relatively easily with soft electrophiles. 5 (2008) Fig.e.104) percent of the conjugate to be formed nonenzymatically..e. . Indeed. Thus. A convenient mechanism accounting for the nonenzymatic reactions of glutathione is based on electrophilicity – nucleophilicity considerations [281].104) or large (Case 3 in Fig. For the definition of hard and soft nucleophilic sites.2274 CHEMISTRY & BIODIVERSITY – Vol. depending on enzymatic efficiency.104). hard electrophilic sites have a highly localized positive charge (i. 4. In the presence of hard electrophiles. In contrast.e. the reader just needs to replace electrophilic with nucleophilic and positive with negative in the two previous sentences. 4. this spontaneous reactivity will allow a small (Case 1 in Fig. a low charge density) which is readily polarizable by the approaching reaction partner.105.. 4. remains highly localized during the approach of the reaction partner). electrophiles and nucleophiles are ranked according to their softness or hardness. soft electrophilic sites have a delocalized positive charge (i. Glutathione (in protonated and ionized form) is a soft nucleophile. spontaneous reaction is precluded (Cases 2 and 4 in Fig. 4. a high charge density) which is poorly polarizable (i..

5 (2008) 2275 Fig. THE major) chemical protection against reactive xenobiotics and reactive compounds produced during the metabolism of endogenous and exogenous compounds. the neat detoxification mechanisms of GSH-GSTs may sometimes backfire and yield reactive metabolites [284]. This will be illustrated in some of the following Figures. The Table shown in this Figure (taken from [281a]) points to the fact that thiols are among the softest endogenous nucleophiles. Examples include products of lipid peroxidation and radiation damage (see also Fig.CHEMISTRY & BIODIVERSITY – Vol. 4. implying that GSH will react preferentially with the soft electrophiles on the right of the Table. 4. Were it not for catalytic facilitation by GSTs. Because things are never simple in nature. Glutathione and glutathione transferases have evolved as a major (in fact.106. . Significantly. GSH would not be able to detoxify the harder electrophiles listed here.98) [282] [283]. these harder electrophiles are precisely the ones that best react spontaneously with various nucleophilic sites in nucleic acids. being thus able to induce DNA damage.



Fig. 4.107. Once formed, glutathione conjugates 4.391 are extensively processed in the body through transport and degradation. Thus, they are substrates of a number of transporters (carriers and export pumps) which mediate their transport across cell membranes in liver, intestine, and kidney, among others organs [285] [286]. More significant in our context is their stepwise enzymatic breakdown to their major excretion products. Their degradation begins with the cleavage of the glutamyl residue by g-glutamyltranspeptidase (EC; human genes GGT1, GGT3, GGT5, and GGT6). These are membranal enzymes present in the liver, kidneys, and other organs. The resulting cysteinylglycine conjugate 4.392 loses its glycyl moiety by the action of various dipeptidases (EC 3.4.13). The cysteine conjugate 4.393 so produced undergoes a reaction of N-acetylation catalyzed by cysteine S-conjugate N-acetyltransferase (EC, a microsomal enzyme found mainly in the kidney. The product is an Nacetylcysteine conjugate known as a mercapturic acid (4.394), a major urinary excretion product of glutathione conjugates. The N-acetylation of cysteine conjugates is reversible due to the involvement of various amidases (EC 3.5.1). More importantly, both the cysteine and the N-acetylcysteine conjugates are substrates of cysteine Sconjugate b-lyase (EC; human genes CCBL1 and CCBL2), a mainly renal and hepatic enzyme which cleaves the SÀC bond in the cysteinyl moiety, thus liberating a thiolated metabolite, 4.395, of the parent xenobiotic [8] [287 – 290]. The latter can be further S-methylated by thiol S-methyltransferase (EC; TMT; see Chapt. 4.2) to form 4.396. Both 4.395 and 4.396 are substrates of cytochromes P450 and flavincontaining monooxygenases which catalyze their S-oxygenation (see Part 2 [2] [4] [291]).



Fig. 4.108. With this Figure, we begin our survey of the various types of substrates and reactions involved in glutathione conjugation. The Sect. 4.7.2 is dedicated to reactions of additions as summarized in Fig. 4.102, whereas reactions of addition – elimination (summarized in Fig. 4.103) will be presented in Sect. 4.7.3. A first class of reactions of addition are on saturated C-atoms, the substrates being the so-called nitrogen mustards (this Figure) and epoxides (Figs. 4.109 – 4.111). Nitrogen mustards are antitumor alkylating agents containing a (2-chloroethyl)amino function, as exemplified here with mechlorethamine (4.397) [292] [293]. Other drugs include chlorambucil, melphalan, cyclophosphamide, and ifosfamide. The global reaction of conjugation may appear as the substitution of a Cl-atom with a glutathionyl moiety, but its mechanism is not a genuine substitution, since spontaneous dechlorination first forms an aziridinium ion, 4.398, which is the actual alkylating species. This reactive intermediate also undergoes enzymatic and nonenzymatic conjugation with GSH, yielding first the conjugate 4.399, then the diconjugate 4.403 from aziridinium 4.401. Some spontaneous hydrolytic dechlorination also occurs to produce the (2-hydroxyethyl)amino derivatives 4.400, 4.404, and 4.405.



Fig. 4.109. Epoxides are conveniently classified into the reactive arene oxides and the more stable alkene oxides (see Parts 2 and 3 [2 – 4]). Numerous drugs and other xenobiotics contain a phenyl moiety whose oxidation yields an arene oxide of generic structure 4.406. Nucleophilic attack by the glutathionyl anion at either of the two oxirane C-atoms occurs with inversion of configuration (i.e., an SN2 mechanism) to yield the glutathionyl conjugate 4.407. In vivo, the latter is processed as presented in Fig. 4.107 to yield the N-acetylcysteinyl conjugate, 4.408, known as a premercapturic acid. Compared to the general case in Fig. 4.107, an additional step is required to produce a mercapturic acid, 4.409, namely aromatization by dehydration [283] [294]. A more detailed presentation of the stereochemistry of the reaction is exemplified with phenanthrene 9,10-epoxide (4.410) [275a] [295]. This substrate is a symmetric meso(R,S)-compound, but GSH attack on either oxirane C-atom cancels the plane of symmetry and renders the products chiral. Due to inversion of configuration at the Catom undergoing substitution, the two metabolites are the (9R,10S)- and (9S,10R)conjugates, 4.411. The product stereoselectivity of the reaction depends on the glutathione S-transferase involved; thus, rat M1-1 gave an approximately equal mixture of the two products, whereas rat M2-2 was essentially stereospecific for the (9R,10S)-conjugate [275a].

418 and (S)-4.417 gave (S)-4.415 and the episulfonium ion 4. especially M3-3. Results with Sprague – Dawley rat liver cytosol were different. since both styrene oxide enantiomers were conjugated predominantly at C(1). The conjugate 4.412.419 in a 1 : 32 ratio.CHEMISTRY & BIODIVERSITY – Vol.3-diene monoepoxide is a chiral compound. . see also Part 3 [3] [4]) is substrate of a number of metabolic pathways including GSH conjugation [296]. This compound is a metabolite of styrene. pure (R)-4. When incubated with liver cytosol from Wistar rats. Substrate enantioselectivity was also observed in that (R)-styrene oxide was conjugated ca. a potential carcinogen.416. Buta-1. This observation was confirmed with rat purified glutathione S-transferases M. namely at C(1) (Reaction a) and at C(2) (Reaction b) [297]. the (R)-enantiomer was conjugated preferentially at C(1).419 in a 6 : 1 ratio. twice as fast as (S)-styrene oxide. 5 (2008) 2279 Fig.3-diene monoepoxide (4.414 resulting from the attack at C(2) proved chemically stable. namely butadiene monoepoxide and styrene oxide. whereas pure (S)-4. The glutathione conjugation of alkene oxides is illustrated here with two industrial examples. namely the thiirane 4. 4. a fast equilibrium was found between conjugate 4. namely butadiene and styrene.413 (resulting from the attack at C(1)) and two thiirane products resulting from an intramolecular rearrangement. Both xenobiotics are oxygenated by CYP2E1 (see Part 2 [2] [4]) to their respective epoxides.418 and (R)-4. In contrast. whereas the (S)-enantiomer was conjugated almost exclusively at C(2).110. yet styrene oxide (4. When incubated with human placental GST. the compound underwent GSH attack at both C-atoms of the oxirane ring. Buta-1.417) appears better suited to discuss product regio.417 gave (R)-4.and stereoselective glutathione conjugation [298]. In other words.

The in vivo metabolism of the COX-2 inhibitor valdecoxib (4. the other with that of an alkene oxide. Another example is simvastatin (4. one dealing with the GSH conjugation of an arene oxide.111. 5 (2008) Fig.107. Two medicinally relevant examples are shown here.423. and finally the various metabolic steps summarized in Fig. revealing a total of 16 metabolites which accounted for practically the totality of its in vivo metabolism [299].420) was examined in great details in mice. . Of relevance here was the methyl sulfone 4.421. glutathione conjugation to 4.181) whose glutathione conjugate 4.5’-epoxy-6’-hydroxy intermediate 4. 4.2280 CHEMISTRY & BIODIVERSITY – Vol. 4.422 whose formation was quite reasonably postulated to have involved phenyl epoxidation.424 was characterized as a major biliary metabolite in rats administered the drug [300]. This conjugate was most probably formed from the 4a’.

b-unsaturated ketones. However. Shown here are three types of a.b-unsaturated aldehydes.425). a. Furthermore. as we shall see later [302]. a. The lower part of the Figure presents a selection of polarized alkenyl moieties known to undergo glutathione conjugation [303].101 using 4-phenylbut-3-en-2-one (4. Some of these moieties are exemplified in the next two Figures. Polarized alkenyl moieties make good targets for enzymatic and nonenzymatic GSH conjugation by a Michael reaction.CHEMISTRY & BIODIVERSITY – Vol.112. .b-unsaturated nitriles. as an example. the reverse reaction (elimination) being also catalyzed by mu class GSTs. and as a rule. a.bunsaturated esters. The initial catalytic step in reaction has been summarized in Fig.bunsaturated ketone. the substrate has two enantiotopic faces such that the saturation of and addition to the C(b)-atom creates a stereogenic center. with the preferential stereoisomer formed being (R)-4. Returning to the same substrate.426 [301]. we note that the reaction is regioselective for the C(b)-atom. two types of a. An important feature of the GSH conjugation of polarized alkenes is its reversible nature. an a. this reversibility is modest and its in vivo relevance small compared with that of the glutathione conjugates of isocyanates and isothiocyanates. Class mu enzymes appear particularly effective in catalyzing the GSH conjugation of phenylbutenone. 4.b-unsaturated amides. 4. and alkenyl groups polarized by two different aryl moieties. 5 (2008) 2281 Fig.

In the case of the epoxide of acrylonitrile. its epoxide formed the two mercapturic acids 4.2282 CHEMISTRY & BIODIVERSITY – Vol. it underwent spontaneous decyanylation before being transformed into the mercapturate 4.432. In the case of acrylamide (4.434. toxic endogenous products of radical reactions and lipid peroxidation such as N-propenal purines and pyrimidines. Both acrylamide and acrylonitrile underwent direct glutathione conjugation which resulted in the excretion of the mercapturates 4.430 which resulted from GSH addition to the C(a). As for the conjugate produced by the attack at C(b). with the attack at C(a) producing the mercapturate 4. namely the two important industrial chemicals acrylamide (4. 4. respectively.431) [304].427) and acrylonitrile (4. Rats administered the two xenobiotics together excreted three mercapturic acids as major urinary metabolites of each chemical.113. Many different classes of chemicals are detoxified by GSH conjugation.433. Here. respectively.427). For both epoxides. 5 (2008) Fig. for example. C(b)-conjugation predominated over C(a)-conjugation. we focus on xenobiotic toxins. and 4-hydroxyalkenals [303a].and C(b)-atom.428 and 4.429 and 4. . the same two positions were targets of GSH conjugation. Furthermore. they both formed the corresponding epoxide.

Morphinone was suspected of being responsible for the rapid.439 spontaneously eliminated 6-thioguanine (4. GST M1-1 and A4-4 were the most active enzymes among the 13 human GSTs examined.436). The hypothesis gained likelihood when the glutathione conjugate of morphinone was unambiguously identified as a biliary metabolite of morphine in guinea pigs. A few examples of medicinal relevance are presented here.114.437). this role being played here by the quinolin-2-yl moiety. Although there was spontaneous GSH addition to verlukast (4. 25% of a dose.437). because it lacks a polarizing carbonyl group. accounting for almost 10% of a dose. the broken arrows pointing to the C-atom attacked by GSH. 4. namely the glutathione-dependent activation of potential prodrugs of cytotoxic thiopurines. dose-dependent decrease in hepatic glutathione content caused by morphine. We begin with morphinone (4. the biliary excretion of the glutathione conjugate and its breakdown products accounted for ca.438) features a butenone promoiety as target for GSH conjugation [308]. 5 (2008) 2283 Fig. In rats in vivo. the reaction was clearly enzyme-catalyzed in rat liver and renal cytosol.440).CHEMISTRY & BIODIVERSITY – Vol. a well-known metabolite. The compound is of interest. A more intriguing example is that of verlukast (4. The transient conjugate 4. Thus. trans-6-[2-acetylethenyl)sulfanyl]guanine (4. Our last example is quite unusual. . The glutathione conjugate of the diuretic drug ethacrynic acid (4.435). is formed both enzymatically and nonenzymatically by attack at the electron-deficient exo-methylidene C-atom [306]. a leukotriene D4 antagonist that was under development for the treatment of bronchial asthma [307]. a rather unstable metabolite of morphine produced by dehydrogenation of its secondary alcohol group [305].

We begin here with natural catechols such as dihydrocaffeic acid (4.450).6-diglutathionyl-TBHQ (4.447. 4. which also proceeds by a mechanism of Michael addition. 4. The latter diconjugate proves that the monoconjugate 4. and the diconjugate 3. the compound was metabolized to three biliary glutathione conjugates. .115. A comparable activation to an ortho-quinone appears to be involved in the neurotoxicity of (methylenedioxy)amphetamine and the infamous ecstasy [311]. TBHQ) [312]. They are also significant in a toxicological perspective. where their glutathione conjugation was alluded to. since it often follows transport of the monoconjugate to target tissues [313].448 is itself a substrate for oxidation – conjugation involving the intermediacy of the glutathionyl-quinone 4.2284 CHEMISTRY & BIODIVERSITY – Vol.446. The formation and reactivity of quinones has been discussed repeatedly in Part 2. When administered to rats. and 4. An example of a para-quinone is provided by the antioxidant 2-(tertbutyl)hydroquinone (4. In biology. 5 (2008) Fig.441). the two monoconjugates formed from the quinone 4. We now take a closer look at this reaction.442 which readily formed the three GSH conjugates 4.449. These are. this compound has been shown to promote kidney and bladder carcinogenicity in the rat. in increasing quantitative importance.443. quinones play important roles in electron transfer and oxygen activation [309].444. At high doses. as the oxidation of some drugs produces quinones or quinone-imines which may form adducts with biomacromolecules (toxification). the former seemingly being the predominant one [310].445.448) and 5-glutathionyl-TBHQ. Oxidation by cytochrome P450 and/or peroxidases yielded the highly reactive ortho-quinone 4. This further sequence of oxidation is particularly significant in a toxicological perspective. namely 6-glutathionyl-TBHQ (4. This capacity for adduct formation is correlated with their reactivity toward glutathione (detoxification) [283a].

451. this risk appears related to the formation of the major metabolite 5-hydroxythiabendazole (4. a feature shared by other reactive intermediates [283a].456.453).43). and it is in direct competition with covalent binding to proteins [280a].457) and to subsequent covalent binding to biomacromolecules. The GSH conjugation of this highly reactive electrophile to form the conjugate 4. In fact.452 proceeds both enzymatically and nonenzymatically. The structure of this intermediate is interesting in that the imino moiety is endocyclic. this aromatic amine was readily oxidized by peroxidases.458 [315]. A more recent finding is that of the toxification of tolcapone (4. whose use is also associated with a risk of nephroand hepatotoxicity.459 whose formation was consistent with the intermediate quinoneimine 4. an inhibitor of catechol O-methyltransferase (COMT) whose therapeutic use is known to be associated with a risk of liver disorders. The classical example of a quinoneimine undergoing glutathione conjugation is that of N-acetyl-para-quinoneimine (4. Microsomal incubations of thiabendazole or its 5-OH metabolite in the presence of GSH afforded the glutathione conjugate 4. A comparable story is seen with the antihelminthic drug thiabendazole.454 as a reduced metabolite of tolcapone [314]. or various CYPs to an intermediate that reacted with glutathione to yield the GSH conjugate 4. 4. see Fig. A potential pathway of toxification has emerged with the discovery of the primary amine 4. Indeed.CHEMISTRY & BIODIVERSITY – Vol. the toxic metabolite of paracetamol (4.455 as the reactive species. .142.116. All evidence points to the orthoquinoneimine 4. 5 (2008) 2285 Fig. human liver microsomes. 4. see Part 2 [2] [4]).

463 had undergone addition at the terminal.460). thereby binding covalently to soluble and macromolecular nucleophiles [316]. 4. These metabolites are strong alkylating agents which undergo Michael additions. but glutathione addition occurs only at ring A to yield the two conjugates 4. The GSH conjugate 4. but may occur at other activated positions in more complex systems. Of further biological and toxicological interest is the oxidation of some 4alkylphenols (or cyclic systems containing a 4-methylidenephenol moiety) to quinone methides.2286 CHEMISTRY & BIODIVERSITY – Vol.117.462 resulted from addition at the exocyclic CH C-atom.465 is the one predominating at neutral pH where quercetin exists as an anion.466 and 4.461 which reacted nonenzymatically with glutathione [317]. . conjugated CH2 C-atom.464). which in the presence of peroxidases is oxidized to a quinonemethide existing as three resonance forms which all react with glutathione. whereas the conjugate 4. Eugenol (4. which occurs at the exocyclic methylidene C-atom in 4-alkylphenols. 5 (2008) Fig. A more complex example is provided by the flavonoid quercetin (4. Like other quinones. these quinonemethides are detoxified by glutathione addition. a main component of oil of clove and an antibacterial agent. This resonance form features two quinonemethide motifs as shown. was oxidized by rat liver and lung microsomes to the quinonemethide 4. The highly delocalized resonance form 4.467 [318].

473.110) can undergo heterolytic cleavage to generate a highly reactive.469. Interestingly. 4.474. The formation of four conjugates (4. the nitrenium ion (4. a final step of dehydration is needed to restore aromaticity and yield 4.118. The presence of the 4a-OH group enhances the electrophilicity of C(4) and explains the regioselective addition of GSH. As shown. C(3). but underwent GSH-dependent reduction as discussed in Sect. . 4.469 was not isolated as such. and 4. the first step being a nucleophilic addition of the HO À anion to C(4a) to yield the quinolimide 4.474 is of particular interest.CHEMISTRY & BIODIVERSITY – Vol.33. An important point to note is that the Nglutathionyl conjugate 4. we saw how the sulfate of N-(9H-fluoren-2-yl)-N-hydroxyacetamide (4. In Fig.472) can be understood as resulting from a direct nucleophilic attack on the electron-poor sites N. and C(7).7. these resonance forms can help explain the nonenzymatic production of several glutathione conjugates observed when 4.468) exists in a number of resonance forms.471. since its formation was shown to be an indirect one. 4.468 was reacted with glutathione [319]. 4. 5 (2008) 2287 Fig. implying its stabilization by extended delocalization of its positive charge. This story was left unfinished and continues here in the context of detoxification by glutathione conjugation.4.470. adductforming nitrenium ion. C(1). The case of the 4-glutathionyl conjugate 4. 4.

resp. 1.477). 5 (2008) Fig.2-trifluoroethene (4. . Haloalkenes are important substrates of glutathione transferases. and 1-bromo-1-chloro-2.3-pentafluoro-2-(fluoromethoxy)prop-1-ene (4.3. i. thus accounting for the nephrotoxicity of compound 4.487.478.3. However.. or react with H2O. The major toxicological interest of both glutathione conjugates is their further biotransformation to reactive metabolites identified or inferred in patients and in rats.2.2288 CHEMISTRY & BIODIVERSITY – Vol.486) to yield the highly reactive thioacyl fluoride 4.e.119.484. 1-chloro-1. The latter can bind covalently to biomacromolecules.483 [323]. undergo blyase-catalyzed cleavage to the thiols 4. it also yields the product of addition – elimination 4. These are reactions of addition – elimination which we will discuss in Sect.481) generated by CO2 absorbents in anesthesia machines.485) or tautomerism (in case of 4. These thiols spontaneously rearrange by loss of HF (in case of 4.476. Their glutathione conjugates (4. the two glutathione conjugates are transformed to cysteine conjugates and mercapturic acids which.480. a small sub-group of haloalkenes. and 4. in analogy with most haloalkenes. tetrafluoroethylene). 4. 4.479). as a 1. a potentially toxic breakdown product of the general anesthetic sevoflurane (4. This breakdown product is a substrate of GSH conjugation. tends to react by glutathione addition rather than by addition – elimination.482).2-difluoroethene (4. in turn.3.482. their conjugation with glutathione involving a mechanism known as nucleophilic vinylic substitution (the SNV reaction) [320 – 322]. An example of some medicinal interest is provided by 1.) result from a reaction of addition which is not followed by elimination. And. This is exemplified here with tetrafluoroethene (4.7.1difluoroalkenes.1. 4.475.486 (see Fig.1-difluoroethene derivative. it does yield the addition conjugate 4.485 and 4. 4. Indeed.107).

4.489 were major metabolites in bile and urine. A simple example is provided by dichloroethyne (4.493 and the highly unsaturated aldehyde 4. The glutathione conjugate 4. and was caused by reactive species such as the thioacyl chloride 4. hemoglobin.e.487.491.CHEMISTRY & BIODIVERSITY – Vol. dichloroacetylene).120. . yielding the glutathione conjugate 4.492). respectively. 5 (2008) 2289 Fig. High covalent binding was seen to albumin. renal DNA. The latter was trapped by GSH added to the incubates. Other metabolites were breakdown products of the thiol 4.490. In rats exposed to low levels of dichloroethyne vapors. acetylenic derivatives) are known to undergo glutathione conjugation. A few alkynes (i. the compound underwent glutathione conjugation as its major metabolic pathway.488 and the mercapturic acid 4. A medicinal example is that of the antifungal agent terbinafine (4. a product of the alkaline decomposition of trichloroethylene and a potent neurotoxin and nephrotoxin [324]. and renal proteins. these substrates have in common a CC bond activated by electron delocalization. When incubated with human or rat liver microsomes.494 [325]. the drug underwent CYP-catalyzed N-dealkylation to the secondary amine 4. This conjugate has thus been postulated to be excreted in the bile where it could cause hepatobiliary dysfunction..495 which retains a second reactive electrophilic site.

As exemplified here with methyl isocyanate (4. as exemplified with N-formylamphetamine (4.503) [331]. the reaction being reversible. Isocyanates also react enzymatically and spontaneously with glutathione. Their main pathway of detoxification is by hydrolysis (Reaction b). eyes. and polyurethanes. 2.2290 CHEMISTRY & BIODIVERSITY – Vol. liberate therein a toxic isocyanate [328].500).496 [327].. whereas the latter undergoes general base-catalyzed cleavage to para-chlorophenyl isocyanate (4. namely a) massive damage to exposed areas (respiratory tract.497 can liberate the parent compound or be processed to cysteine conjugates (e.121. The former is oxidized by CYPs to the isocyanate 4. Because glutathione conjugates are substrates of various transporters.4diisocyanatotoluene (4. as exemplified by the effects of the infamous methyl isocyanate [326]. but the amine so liberated may produce hypersensitivity [107]. Organic isocyanates (RÀN ¼ C¼O) are used extensively to manufacture paints. they may reach deep compartments in the body and. etc. The lower part of the Figure presents other examples. and b) hypersensitivity of the respiratory tract. 4.499) is another chemical widely used in the paint and plastic industries. a contaminant in illicit preparations of amphetamine analogues. if the conjugation is reversible. In other words.502). Thus. and the antitumor agent sulofenur (4. .498) which also liberates 4. the mercapturate 4. GSH conjugates of isocyanates are transport forms as much as products of detoxification. the toxicity of isocyanates is a consequence of their reactivity as electrophiles toward biomacromolecules such as proteins (Reaction a).g.) in the case of heavy accidental exposure.501 [330]. At the molecular level. 5 (2008) Fig.496). Their reactivity can explain toxic reactions. Isocyanates may also be formed as metabolites of xenobiotics. pesticides. as seen in workers exposed to polluted atmospheres. its glutathione conjugate 4. and a cause of occupational asthma [329].

and the conjugate 4. The mercapturate metabolites of sulforaphane (4. A major route was sulfoxide reduction to erucin (4. and P1-1 were shown to be catalytically competent.505).511). Other natural isothiocyanates known to undergo enzymatic and nonenzymatic glutathione conjugation are allyl isothiocyanate (4. 5 (2008) 2291 Fig. respectively. cabbage.505) were the major urinary metabolites.507 and 4. One such compound is sulforaphane (4. The metabolism of these natural products is of interest due to their chemoprotective properties. 4. A number of edible cruciferous vegetables such as broccoli.504) underwent both redox and conjugation reactions. .504) and erucin (4. and cauliflower contain organic isothiocyanates known to induce various glutathione Stransferases. As shown here. with the forward reaction being markedly faster than the reverse one.e.509. i.510). The GSTs A1-1. M1a-1a. Like that of isocyanates. and phenethyl isothiocyanate (4. namely the glutathionyl conjugates of erucin and sulforaphane.504).508.122. 4. the glutathione conjugation of isothiocyanates is reversible. A2-2.512) [333].506. whereas a minor pathway was dehydrogenation to 4. sulforaphane (4. Three metabolites were identified in the bile. benzyl isothiocyanate (4.CHEMISTRY & BIODIVERSITY – Vol. the metabolism of which was carefully investigated in rats [332]..

2292 CHEMISTRY & BIODIVERSITY – Vol. 4.513) [335].515). In a toxicological perspective. reactive enough to form DNA adducts. whereas the (S)enantiomer gave the (R)-4. for example. In the presence of rat liver cytosol. 4. 5 (2008) Fig. . a nucleophilic substitution with inversion of configuration.517 to methanol (4. it is informative to examine the conjugation of dichloromethane (4. 4. namely chloroalkanes and bromoalkanes [334]. Their conjugation with glutathione occurs according to an SN2 mechanism. glutathione conjugation was substrate-enantioselective in that the (R)-enantiomer was a better substrate of GSTs than the (S)-enantiomer. a type of reaction to be discussed in Sect.123. In a first part. Another reaction worthy of mention is the GSH-dependent reduction of 4. in other words. and we begin with simple haloalkanes.516. with the model compound 2-bromo-3-phenylpropanoic acid (4. itself an unstable metabolite which loses GSH to yield the toxic formaldehyde (4.118). better designated as nucleophilic addition – elimination. Nucleophilic substitution of the Clatom yields a conjugate.7.514 conjugate. The reaction was also product-stereoselective in that the (R)enantiomer was metabolized to the epimeric (S)-4. 4.519).517.518).4 (see also Fig.514 conjugate. we shall examine conjugation at sp3-C-atoms. This Section presents the major reactions of glutathione conjugation involving nucleophilic substitution. a compound representative of toxic halomethanes [336] [337]. This metabolite can undergo nonenzymatic hydrolytic dechlorination to 4. This was nicely shown in a number of studies.

525.e.2-dihaloethyl motif are another class of xenobiotics able to undergo GSH/GST-catalyzed toxification [338]. Haloalkanes containing a 1. 4. cyclization to the episulfonium species 4.531 and 4.530. i. or by formation of the diglutathionyl conjugate 4.527.521). and reduction to the diol conjugate 4. in turn. 4. respectively.528 and 4.532.2-dibromo-3-chloropropane (4. 4.526 [339]. respectively. However. 4. and 4.CHEMISTRY & BIODIVERSITY – Vol. The nematocide 1.522. form an episulfonium ion. two among the three resulting metabolites. The second pathway is the direct. GST-catalyzed formation of the glutathione conjugate 4.520) offers an eloquent example of alternating steps of toxification and detoxification characteristic of the metabolism of numerous xenobiotics.525.. followed by GSH conjugation to 4. This fact is indicated in the Figure by the red boxes in which the episulfonium formulae are embedded. The latter is unstable by virtue of its vicinal halide-glutathione motif and undergoes spontaneous debromination to form an intermediate episulfonium species. yielding metabolites 4. 5 (2008) 2293 Fig.124.534. hydrolytic ring opening of the three-membered (thiirane) ring to 4.524.529 and 4.523. Two major metabolic pathways lead to its toxification. able to bind covalently to DNA. . The compound is a persistent environmental pollutant that was used as a soil fumigant. retain a vicinal halide-glutathione motif and can. This toxic species can also be detoxified by hydrolytic ring opening to 4. The first pathway is a CYP-catalyzed oxidation to 2-bromoacrolein (4. 4. These routes are followed by another round of detoxification by hydrolysis or GSH conjugation.529.530.533.

. 4. The benzylic C-atom in arylmethanol sulfates is strongly activated by the electron-withdrawing O-sulfate moiety.541. 4. 4. diallyl sulfoxide (4. The same mechanism was presented in Figs. an irreversible proton pump inhibitor used to treat peptic ulcers and other related diseases [341].30 – 4. but 4.2294 CHEMISTRY & BIODIVERSITY – Vol.540 formed a glutathionyl disulfide.535). Besides haloalkanes and arylmethanol sulfates.537) reacted spontaneously with glutathione to form the GSH conjugate 4. the C(2)-atom of benzimidazole was also found to be a site of GSH addition – elimination according to a mechanism discussed below (see Fig.125. Diallyl sulfide (4. a few other types of substituents facilitate GSH conjugation by addition – elimination at sp3-C-atoms.538. 5 (2008) Fig. In vitro. a flavor component of garlic. As shown with Reaction a. Rats dosed with 4. according to a reaction discussed in Fig. 4. the benzylic position is activated by the S¼O group and is a good target for glutathione conjugation.540.542).539 and 4. is considered a chemoprotective agent due to its inhibition of CYP2E1-toxification of some carcinogenic chemicals. Our next example is a medicinal one. Note that neither of the two GSH conjugates 4. Interestingly.536) and diallyl sulfone (4.130. 4. The moieties eliminated. notably sulfoxide and sulfone groups as exemplified here. 4. were not characterized as such.3). respectively.127).535 excreted in their bile a variety of metabolites resulting from initial sulfoxidation and glutathione conjugation [340].32 (Chapt.543 and 4. 4. where we saw how reactive arylmethanol sulfates are detoxified by glutathione conjugation. as seen in rats dosed with the drug. namely pantoprazole (4.544 was identified as such but as further breakdown products. Halide atoms in haloalkanes are electron-withdrawing substituents which activate (render electrophilic) these compounds toward GSH conjugation.

547 and 4.550 and the thioketene 4.1.4-hexachlorobuta-1.551 is also well known. . 5 (2008) 2295 Fig.126. Haloalkenes of relevance here include the much studied 1. haloalkenes tend to react with GSH/GSTs in a reaction of addition – elimination whose mechanism is a nucleophilic vinyl substitution (SNV) [320] [321] [342]. since it appears to be metabolized in vivo exclusively by glutathione conjugation [320]. and its loss of HCl to form the thioketene 4. 4. This thiol has an intrinsic reactivity which favors its isomerization to the thioacyl chloride 4.552 (and to the corresponding carboxylic acid).545). detoxification of the thioacyl chloride 4.549. it is then degraded to the cysteinyl and mercapturate conjugates. respectively. 4.2. They undergo CYP-catalyzed oxidation and GST-catalyzed conjugation.2-trichloroethylene [343] [344]. particularly by hydrolysis to the thio O-acid 4.119). 4. The latter is preferentially excreted renally.550. we focus on 1.1.4. Both are highly reactive metabolites which react with nucleic acids and proteins.2.CHEMISTRY & BIODIVERSITY – Vol. in close analogy with the reaction of aromatic substitution to be presented in the next Figure.1. However.548.1-difluoroalkenes (see Fig. is formed at C(1). the enzymes in the latter case being mainly microsomal glutathione S-transferases [319]. However.546.2-tetrachloroethylene and 1. while the former is a preferred substrate of b-lyase to yield the thiol metabolite 4. 4. With the exception of 1. Haloalkenes have a number of industrial uses and are also environmental contaminants. As shown.3-diene (4. thereby accounting for the toxicity and carcinogenicity of hexachlorobutadiene and other haloalkenes. This mechanism involves a reaction intermediate where the target C-atom is in an sp3-configuration.551. the glutathione conjugate of hexachlorobutadiene.3.

4.554 will also depend on the relative ease of elimination of the substituent.. A similar reasoning applies to chlorpyrifos (4. In both cases. 4. except among other things for a greater number of activating and displaceable groups. the geminal N-atom and an ortho-Cl substituent delocalize the negative charge. the formation of the s-complex being reversible. A number of chloro.555) where any of the three F-atoms can be displaced [345]. e. see also Fig. aryl sulfones.101) begins with the formation of a s-complex (also known as a Mesenheimer complex) [275a]. Addition – elimination reactions of glutathione to substituted aromatic rings are comparable to those targeting haloalkenes.558).3.127. The overall rate of formation of the GSH conjugate 4.4-dinitrobenzene (4.2296 CHEMISTRY & BIODIVERSITY – Vol. A1-1. 5 (2008) Fig.g. The mechanism of the reaction. A number of human GSTs catalyze the reaction. 4. .g.. and arylsulfonamides [346]. A2-2.and para-substituents.557) may appear unexpected until one realizes that a scomplex at C(4) has its negative charge delocalized to the three neighboring N-atoms [347]. and P1-1.560).5-trifluoro-2-nitrobenzene (4. The better the delocalization of the negative charge to electron-withdrawing ortho.553.and fluoroarenes are known to be substrates of the reaction. a widely used pesticide to which many humans and animals are exposed [348]. M1a-1a. Other substrates of the reaction include aryl sulfoxides (e. The GSH conjugation of the antihypertensive agent moxonidine (4. as exemplified here with 1-chloro-2. This xenobiotic is conjugated by GSTs at C(6) (loss of the Cl À anion to yield conjugate 4.556). the easier is the formation of the complex. for example 1.559) and C(2) (loss of the diethyl thiophosphate moiety to yield 4.

564. it would imply a reaction of addition – elimination resembling the GSH conjugation of both haloalkenes and haloarenes. namely its hydroquinone derivative 4.565. assuming its formation indeed involved the postulated pathway. The latter was oxidized further to a quinone before undergoing cyclization (not shown).566. This atypical neuroleptic was withdrawn in 1993 due to a few cases of aplastic anemia observed in patients receiving the drug. When incubated with stimulated human neutrophil granulocytes. followed by a GSH Michael addition to the diglutathionyl conjugate 4.561) [349]. . A seemingly special case of glutathione conjugation has been reported for remoxipride (4.563.CHEMISTRY & BIODIVERSITY – Vol. Based on in vitro studies.564 was substrate of a second round of oxidation to 4. which reacted with glutathione to form the glutathionyl conjugate 4. this metabolite underwent peroxidase-catalyzed oxidation to the reactive quinone 4.128. 5 (2008) 2297 Fig.562. the reactive quinones detected in this study bring convincing evidence for multiple peroxidase-catalyzed toxifications alternating with glutathione conjugations. The monoglutathionyl conjugate 4. a potential mechanism of toxification was deduced which implicated a known human metabolite of the drug. 4. The formation of this metabolite was demonstrated unambiguously. In summary.

Its major route of toxification in human lung and liver microsomes was found to involve CYP2E1 oxidation to the epoxide 4. Other activated acyl derivatives of distinct relevance in xenobiotic metabolism are acyl glucuronides (see Chapt.4) and acyl-coenzyme A conjugates (see Chapt.575 [351].1-dichloroethylene) [350].567.568. 5 (2008) Fig.1-dichloroethene (4. The latter reacted (presumably spontaneously) with GSH to form (S-glutathionyl)acetyl glutathione (4.571). and they tend to react enzymatically and/or spontaneously with glutathione. 1. 4. 4. followed by GSH addition and HCl elimination to generate the adduct-forming (S-glutathionyl)acetyl chloride (4. Activated derivatives of carboxylic acids can behave as acylating agents toward nucleophiles.570). An interesting example of such a compound has been reported as a metabolite of 1. both of which are intermediates in the in vivo and in vitro formation of clofibryl-S-acyl-glutathione 4.6). and with water to form S-glutathionyl acetate (4.50 and 4. the lipid-lowering agent clofibric acid (4. This chemical is extensively used in the manufacture of plastics and is known to be a lung and liver toxicant.572) forms an acyl-CoA conjugate (4.2298 CHEMISTRY & BIODIVERSITY – Vol. both being able to form glutathione conjugates in a reaction of transacylation (Figs. . Thus.569). 4. 4. and particularly acyl chlorides used extensively in organic chemistry.573) and an acyl glucuronide 4. Acyl halides among the most reactive among such derivatives.574.129.83).

However. One mechanism for their formation is by reaction of the glutathionyl radical with another thiyl radical. A classical example is that of the antimineral corticoid (diuretic) drug spironolactone (4. 4. was trapped by GSH to form the glutathionyl-spironolactone disulfide 4. 5 (2008) 2299 Fig.130. 4.98). 4. a major pathway of which is hydrolysis of the thioacetate ester group to the thiol metabolite 4. in analogy with the formation of oxidized glutathione (GSSG.579 [352]. followed by S-methylation (Chapt. this mechanism does not appear to be effective compared to the xenobiotic thiol being oxidized to a sulfenic acid (RÀSÀOH) prior to reacting with GSH.576). .CHEMISTRY & BIODIVERSITY – Vol. when leaving the enzyme catalytic site.2) to the thiomethyl derivative 4. In addition. The compound has a complex metabolic fate.580 according to the mechanism shown. This reactive metabolite. Another reaction of GSH conjugation that can formally be considered as an addition – elimination is the formation of mixed glutathionyl disulfides. the thiol 4.577 is oxidized by CYP and/or flavin-containing monooxygenases to the sulfenic acid 4. a significant human metabolite.578. see Fig.577.

with pi-class GST catalyzing the forward reaction.121). as exemplified by the well-known angiotensin-converting enzyme inhibitor captopril which. Another GSH conjugate (not shown) was the product of addition to the isocyanate group (see Fig. A more unexpected example of a mixed glutathionyl disulfide is that of the antidiabetic troglitazone (4. This is a reversible post-translational modification of low-pKa cysteinyl residues in proteins. The isocyanate group in the latter is broken down by hydration and loss of CO2 . The process plays a role in governing how cells respond to oxidative stress. forms a disulfide bond with human serum albumin [355]. 5 (2008) Fig.2300 CHEMISTRY & BIODIVERSITY – Vol.582) to form the intermediate sulfenic acid 4. 4.131.581) whose use has been associated with cases of hepatotoxicity. in vivo. One pathway in its metabolic fate is scission of the thiazolidinedione ring via an unstable S-oxide (4. The formation of this metabolite was characterized unambiguously in human liver microsomes containing GSH. witness the process of Sglutathionylation [354].584. 4. we note that cysteinyl groups in accessible proteins may also bind some xenobiotic thiols.583 [353]. . To broaden the perspective further. Note that this reaction of glutathione with other thiols is not limited to xenobiotics. whereas the sulfenic group accounts for the formation of the glutathionyl disulfide conjugate 4.

Three such metals will be exemplified here. Arsenic salts react spontaneously with glutathione in solution. thiolates can serve as transport forms of metals in the organism [356].592) excreted the diglutathionyl conjugate 4. As such. The bond between cationic Hg. the formation of thiolates is favored by their low energy.66) which formed the diglutathionyl complex 4.587 also reacts rapidly with Cys-Gly.591) and its analog trimelarsan (4. mercury.132. as seen with methylarsonite (4. platinum. explaining why their conjugation with GSH is a reaction of substitution.590 [43] [47] [359]. Metallic cations forming thiolates include arsenic. forming thiolates (previously known as mercaptides) which are thermodynamically stable but kinetically labile. and silver. Cationic Hg is well-known for its strong affinity for thiols.20 and 4. it has a high affinity for endogenous thiol groups. and is for example highly bound to hemoglobin Cys13a in rats [358]. rats dosed with melarsoprol (4. As for arsenic.589 was the main biliary metabolite in rats dosed with 4. In other words. Like Hg.588) [357]. 5 (2008) 2301 Fig. and the conjugate 4. but the equilibrium constant is also markedly dependent on the relative concentrations of the competing thiol ligands. namely mercury.CHEMISTRY & BIODIVERSITY – Vol.593 in their bile [360]. copper. Compound 4. 4. 4.587. arsenic.585) reacts spontaneously with glutathione to form mercurydiglutathione (4. Thus. whereas methylmercury chloride (4. Arsenical drugs used to treat some parasitic infections are also conjugated by GSH. . or Pt and their counterions is more covalent than ionic ( < 30%). gold.587) forms methylmercuryglutathione (4. it is a worldwide natural contaminant and an occupational hazard (see Figs. cadmium. Thus. and platinum. mercury chloride (4.586). lead. As. also known as mercaptans.21). Glutathione reacts with a number of metallic cations.

The kinetics of reaction of PtII complexes with glutathione [362]. and the role of transporters is slow to emerge.596. to be progressively replaced by a 1 : 2 complex 4. The strength of the SH ligand is also obvious in complexes between cisplatin and N-acetylcysteine incubated in a 1 : 1 molar ratio.599 having the S-atom bridging two Pt-atoms.597. 5 (2008) Fig.594) as an example. a 1 : 1 complex 4. At a GSH/cisplatin molar ratio of 1 : 1. While a global picture integrating adduct formation with DNA.598 was initially formed. biotransformation. Taking the archetypal drug cisplatin (4. Here again. the 1 : 1 complex 4.596 was formed first.133. Platinum complexes have considerable significance in cancer chemotherapy [361]. but decreased as the concentration of the 1 : 2 complex 4.595 was formed. to be progressively replaced by the 1 : 2 GS/Pt complex 4. 4. The toxicity of the 2 : 1 GS/Pt complex was demonstrated in a cell-free system where it inhibited protein synthesis.597 increased. for example. These complexes indicate that the strongest Pt ligand in GSH is indeed its thiol group.2302 CHEMISTRY & BIODIVERSITY – Vol. . that GS – platinum complexes may be excreted from cells by efflux pumps. it appears. as well as their hydration [3] [4] play a significant role in their efficacy and toxicity. Solutions with a GSH/cisplatin molar ratio of 1 : 1 had a cytotoxicity that increased with increasing concentration of the 1 : 1 complex 4. followed by an amino and an amido group. At GSH/cisplatin molar ratios equal or larger than 2 : 1. the 2 : 1 complex 4. and its transport across tumor cell membranes was ATP-dependent. we review here its major glutathione conjugates as formed nonenzymatically in solution under physiological conditions of pH and temperature [363].

2’-dichloroacetophenone (4. The first conjugation step was catalyzed by GSTs A1-1. and P1-1. 4.134. .602). once coupled to a glutathionyl moiety.604 reacts rapidly with a second glutathione thereby reducing the initial hydroperoxide to an alcohol. whose presence was confirmed in human liver mitochondria [366]. Heterolytic cleavage of the YÀS bond yields a molecule of GSSG and the RÀY À anion.600) were investigated as substrates for their GSH-dependent reduction [364]. Second. Here. GSH attacks the proximal Oatom. Two classes of substrates are exemplified in this Figure. namely a-halo ketones and hydroperoxides. 5 (2008) 2303 Fig. glutathione conjugates react with a molecule of glutathione to yield glutathione disulfide (GSSG) and the reduced substrate. Our second example is of great physiopathological significance. Several 2-chloroacetophenones such as 2. A2-2. 4. Glutathione addition and chloride elimination yielded a GS-conjugate (4. These compounds are reactive products of lipid peroxidation as well as intermediates in the synthesis of some prostaglandins [152] [365].601) in which electronic delocalization activated the S-atom. since it implies the reduction of hydroperoxides 4. the target atom Y. must be able to activate the S-atom toward attack by the second glutathione molecule.CHEMISTRY & BIODIVERSITY – Vol.605. First. The heterolytic cleavage of the CH2 ÀSG bond allowed reduction of the substrate to 2’-chloroacetophenone (4. but the resulting intermediate 4.603. The structural and electronic conditions for such a reaction are schematized in the upper part of the Figure using a generic RÀYÀX substrate. In a number of cases. the group X (often a leaving group) must be a good electroattractor able to activate the target atom Y toward GSH attack. The initial conjugation step was catalyzed by GST O1-1.

4. 4. It involves an intramolecular. The same is true for Pathway c. followed by autoxidation or peroxidase-catalyzed oxidation. The reaction.607. Our last case is that of AsV salts (see also As methylation under Sect. 5 (2008) Fig.606. Nitrosoarenes. This pathway is favored at relatively high GSH concentrations and for nitrosoarenes with electron-withdrawing substituents.611 then undergoes two GSH-reduction steps producing two GSSG molecules. Pathway a is a reductive thiolytic cleavage (Fig.4). 4. or b) by reductasecatalyzed reduction of nitroarenes [3] [4]. Pathway b is favored by lower pH and relatively low GSH concentrations and for nitrosoarenes with electron-donating groups. Nitrosoarenes react readily and nonenzymatically with glutathione to form semimercaptals 4. 4. 4. proton-catalyzed rearrangement of the semimercaptal 4.609 to the more stable sulfinanilide 4. yields an intermediate GSH conjugate 4.607.606 and glutathione sulfinic acid.134) produces the AsIII species dimethylarsinous acid (4. 4. 4.2304 CHEMISTRY & BIODIVERSITY – Vol.609 [367] [368].67) [47] [48] [359]. These are metabolic crossroads to three distinct reduction pathways (a.135.2. to aromatic hydroxylamines.610 which tends to undergo proton-catalyzed hydrolysis to the aromatic amine 4. The resulting mercaptal 4. .608. They react enzymatically (GST O1-1) and nonenzymatically with glutathione as exemplified here by dimethylarsinic acid (4. b.134). and c) whose relative significance depends on experimental conditions (GSH concentration and pH) and substrate properties. occur as industrial chemicals or as metabolites formed a) by CYP-catalyzed oxidation of aromatic amines. which is one of addition with elimination of a H2O molecule. which may occur when excess GSH is present. In effect.68). Pathway b is a reduction of a nitrosoarene to an arylamine. its product is an N-arylhydroxylamine 4.612 whose reductive thiolytic cleavage (Fig.

respectively. whereas the former (4.282). at best a very minor and occasional metabolite.5). A well-known example is that of the antituberculosis drug isoniazid (4. the hydrolysis of the latter two hydrazones in biological media (buffer or plasma) was practically negligible. namely the acetone hydrazone 4. Interestingly. The first type of reaction is covered in the present Figure and involves the condensation of xenobiotic hydrazines and hydrazides with an endogenous ketone or aldehyde to form a Schiff base called a hydrazone. These are important in vivo metabolites of isoniazid. which. 4.284).615. especially in slow acetylators [191]. A second relevant example is that of hydralazine (4. Hydrolysis was even faster for the acetaldehyde hydrazone. and the 2oxoglutaric acid hydrazone 4. besides acetylation (see Chapt. we examine two unusual types of metabolic reactions.614. 4.136. namely the nonenzymatic coupling of xenobiotic amines with endogenous carbonyl compounds [369].613 and 4. In this last and short Chapter. Three such conjugates are formed in humans.615) did regenerate the parent drug [370]. the pyruvic acid hydrazone 4. 5 (2008) 2305 Fig.CHEMISTRY & BIODIVERSITY – Vol. again a substrate of Nacetylation which forms hydrazones.617 [190]. . forms hydrazones with circulating pyruvic acid and 2oxoglutaric acid to yield the conjugates 4.616.

From the examples in the literature. As shown for 2-haloethylamines such as 2-bromoethylamine (4.623 which was an important urinary metabolite in humans. The identification of carbamic acids under physiological conditions is often prevented by this lack of stability. meaning that the amine must be basic enough to have a well localized doublet of electrons. a reaction greatly facilitated by the high level of endogenous bicarbonate in human blood (around 20 mm). Bicarbonate ions react with amines to form carbamic acids.137.619 in our example) was unstable and easily hydrolyzed back to the free amine. Several of these carbamic acids would rapidly vanish by hydrolysis and remain unidentified.622 was not detected. 4. on the stability of the carbamic acid.2306 CHEMISTRY & BIODIVERSITY – Vol. 8 is favorable for a reaction under physiological conditions.621). 5 (2008) Fig. a reversible reaction whose rate constants and equilibrium constant depend largely on the reactivity of the amine. The carbamic acid thus formed (4. which served as evidence of the intermediacy of the carbamic acid [372]. However. We illustrate this pathway with mexiletine (4. Three other . Its carbamic acid derivative 4. and on external conditions such as pH and concentrations [371]. were it not for their capacity to serve as substrates of UDP-glucuronosyltransferases and yield carbonyloxy-b-d-glucuronides [373]. an orally effective antiarrhythmic agent [374]. yet not too basic for the unprotonated species to be present in sufficient proportion.619 reacted intramolecularly to form the stable oxazolidin-2-one 4.620.618). in contrast to its carbamoyl glucuronide 4. the reaction involves a nucleophilic attack by the unprotonated amine. it appears that a pKa value of ca. A noteworthy number of medicinal amines are known to form carbamic acids in vivo. the carbamic acid 4.

Hines. D. 5 (2008) 2307 medicinal examples (secondary amines. Biochem. Eds. in ADME-Tox Approaches. Chem. K. L. [11] E. Hydrolyses. www. Liljas. p. 1917 – 1924. Nature 1994.ebi. Jeong. in Burgers Medicinal Chemistry and Drug Discovery. Eds. R. A. 455 – 468. Craddock. Exp. The Biochemistry of Drug Metabolism: Principles. 6th edn. Mouse liver nicotinamide N-methyltransferase. 1049 – Ther. T. Int. 6. The antimicrobial agent garenoxacin (4. [5] G. Elsevier. [15] J. p. Elsevier. Krämer. Drug Crystal structure of catechol O-methyltransferase. W. and Wiley-VCH. S. and species variations. 167 – 191. H. Testa. R. D. . Savolainen. [9] J. Methylation pharmacogenetics: catechol O-methyltransferase. Szumlanski. 24. J. Eur.. Salminen. Part 3: Reactions of Hydrolysis and Their Enzymes. [13] R. Rettie. 2007. Taylor. [10] D. 2003.625) was excreted mostly unchanged in experimental animals. Soine. and histamine N-methyltransferase. Otterness. J. Tenhunen. L. Testa.CHEMISTRY & BIODIVERSITY – Vol. McCarver. Drug Metab. Vol. Biochem. in fact) are shown. The Biochemistry of Drug Metabolism – An Introduction. 2008. van de Waterbeemd. Testa. Oxford. B. Verlag Helvetica Chimica Acta. J. Biochem. Part 2: Redox Reactions and Their H. D. p. The ESTHER Database. Liu. M. ExPASy Proteomics Server. [12] a) M. Genomic organization of the human catechol O-methyltransferase gene and its expression from two distinct promoters. 5 in Comprehensive Medicinal Chemistry. The ontogeny of human drug-metabolizing enzymes: Phase II conjugation enzymes and regulatory mechanisms. Testa. [4] Triggle. B. Ed.brenda-enzymes. 1994. Curr. Vol. Enzyme Nomenclature. Yan. Mulder. 2007. Krämer. 257 – 405. R. 300. Mammalian small molecule methyltransferases: their structural and functional features. 354 – 358.expasy. 2nd edn. S. Krämer. www. Principles of drug metabolism. S. Annu. thiopurine methyltransferase. www. The Biochemistry of Drug Metabolism – An Introduction. Weinshilboum. D. 2nd edn. 19 – 52. J. J. [7] R. 5 in Comprehensive Medicinal Chemistry. 1999. Principles of drug metabolism 3: Enzymes and tissues. 2007. T. London. Coupling of conjugating enzymes and efflux transporters: Impact on bioavailability and drug interactions. Abraham. in ADME-Tox Approaches. van de Waterbeemd. Otterness. Taylor & Francis. 2007. Conjugation reactions in foreign-compound metabolism: definition. 431 – 498. Wiley-Interscience. L. D. J.chem. consequences. 1139 – 1149. Pharmacol. www. A. 133 – 166. Eds. 368. S. D. 1990. b) L. Krämer. Hu. D. G. [8] Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB). Vidgren.626). Toxicol. Testa.inra. Biodivers. REFERENCES [1] B. 39. D. Pharmacol. As for the antidepressant sertraline (4. varenicline (4. Redox Reactions. Ed. Pharmacol. its carbamoyl glucuronide was a major metabolite in dog bile and was also found in the plasma of humans dosed with the drug [377]. J. N. The Biochemistry of Drug Metabolism – An Introduction. B. Thus. Kiviluoto. Chen. J. [2] B. X. Brenda: The Comprehensive Enzyme Information System. 2.. Enzyme Structure Database. Caldwell. I. J. M. 2002. 2005. C. Ulmanen. http://bioweb. Rev. [3] B. [6] B. Principles of drug metabolism 2: Hydrolysis and conjugation reactions. Oxford. Part 1: Principles and Overview. with carboxylic glucuronidation and carbamoyl glucuronidation as minor pathways [376]. Taylor. J. 1997. J. 54. 1053 – 1101. Fujioka. 745 – 777. B.624) is a partial nicotinic receptor agonist whose carbamoyl glucuronidation was catalyzed by UGT2B7 in human liver preparations [375]. Weinshilboum.. A. Testa. Lundstrçm. M. Testa. Chem. Eds. 4. 361 – 366.ensam. 1982. Chem. Weinheim. Zurich. 2031 – 2122. Totah. [14] J. 2006. Svensson. 13. 223. 3. 1992.. B. Conjugation Reactions in Drug Metabolism. M. Biodivers. 4. E. Testa. Triggle. A. M. Hoboken. Vol.qmul. Biodivers.

A. Stack. K. M. [27] G. Maret. A. Prakash. Creveling. 1024 – 1030. excretion. S. Nagata. Julkunen. El Tayar. Exp.  Human catechol O-methyltransferase pharmacogenetics: description of a functional polymorphism and its potential application to neuropsychiatric disorders. Drug Metab. E. T. A. H. Kuo. Matsuura. Role of quinoids in estrogen carcinogenesis. The MPTP story: MAO activates tetrahydropyridine derivatives to toxins causing parkinsonism. S. Skinner. Cavalieri. Holland. 28. Dispos. 2005. 2003. Cui. [22] D. Pharmacol. Conney. Metabolism. 243 – 250. Potchoiba. Lambert. Yu. K. Toxicol. Saiko. Jaw-Tsai. Formation of quaternary amines by N-methylation of azaheterocycles with homogenous amine N-methyltransferase. Testa. Biochem. Y. pharmacokinetics. X. Ther. 35. D. 31. E. Drug Metab. Yang. [20] R. Papolos. F. estradiol-2. F. Inhibition of human liver catechol-O-methyltransferase by tea catechins and their metabolites: structure-activity relationship and molecular-modeling studies. [24] L. H. Chirality 1999. 2000. P. A. Wang. 37. U. 161. Kadlubar. 31. W. 851 – 859. Naoi. J. Proc. and excretion of a highly selective N-methyl-d-aspartate receptor antagonist. Taskinen. J. C. A N-methyltransferase in human brain catalyses N-methylation of 1. M. Carrupt. M. Acad. 34. M. S. . Taskinen. 2003. Metabolism. Nagatsu. I. 160 – 165. [28] a) M. 1350 – 1364. Lu. Toxicol. F. M. Pisha.2. E. Toxicol. Thomas. Saeed. R. J. Woesle. Piche R.2308 CHEMISTRY & BIODIVERSITY – Vol. Gross. ´. E. K. Drug Metab. 1998. I. L. Biophys.3. Qiu. 2004. B. b) D. Thakker. I. R. A. Enzymology of methylation of tea catechins and inhibition of catechol-O-methyltransferase by (À)-epigallocatechin gallate. N-methylisoquinolinium ion. Ansher. Chem. 59. C. Lachman. [21] B. J. F. Res. Fielding. 6. Drug Metab. Jakoby. S. Boehlert. B. 1523 – 1531. 2514 – 2517. Drug Metab. X. Molecular characteristics of catechol estrogen quinones in reactions with deoxyribonucleosides. Res. Dispos. Pharmacol. C. Yang. Mol. E. Kuan. J. Patel. Ansher. J. E. L. 76 – 87. 2006. 1986. 1990. Natl. Molecular mechanisms controlling the rate and specificity of catechol O-methylation by human soluble catechol O-methyltransferase. [26] P. Dubey. Stereospecific Nmethylation of the tetrahydroisoquinoline alkaloids isosalsoline and salsolidine by amine Nmethyltransferase A from bovine liver. T. J. 292 – 299. 2-Hydroxyestradiol is a prodrug of 2-methoxyestradiol. b) A. T. Gillespie. 1113 – 1127. E. Chen. J. M. H. C. 31. Lotta. M. Commun.4-quinone vs. L. L. Y. T. Bolton. Antkowiak. M. Res. 572 – 579. [19] K. E. Byun. 1996. Sci. N. T. C. distribution and excretion of a selective N-methyl-d-aspartate receptor antagonist. C. K. C. T. Zahid.4-tetrahydroisoquinoline into N-methyl-1. F. A. 22. W. 1988. 309. Crooks. Pharmacol. Y. Lautala. Biol. Butler. Prakash. 1988. 5 (2008) ´ [16] T. S. J. Weinshilboum. Regioselectivity of catechol O-methyltransferase. Jenner. S. traxoprodil. The greater reactivity of estradiol-3. Cai. Welsh. Zhang. H. Drug Metab. K. 291 – 332. Damani. 269. Ezell. Jackson. Kirk. T. N. Biochemistry 1995. S. Res. Allison. Chem. Ulmanen. C. M. R. N-Methylation: potential mechanism for metabolic activation of carcinogenic primary arylamines. J. L. [17] H.2. J. Knadler. [18] a) P. 261. 2007. Cavalieri. G. Lantz. E. S. Melen. Liehr. Metabolism.A. Chem. a precursor of a dopaminergic neurotoxin. Johnson. Biochem. 1673 – 1677. W. D. B. 1142 – 1150. 1093 – 1097. C. 11. Bahnmaier. Chem. J. C. Biochem. [25] D. and pharmacokinetics of duloxetine in healthy human subjects. Rogan. 2003. 85. in human cytochrome P450 2D6 extensive and poor metabolizers. Jakoby. D. L. M. Rev. O-Methylation of tea polyphenols catalyzed by human placental cytosolic catechol-O-methyltransferase.3-quinone with DNA in the formation of depurinating adducts: implications for tumor-initiating activity. traxoprodil. 9. 1213 – 1219. Ulmanen. Meng. B. 19. C. Pharmacogenetics 1996. 1989. G. Shah. Godin. H. Zacharia.4-tetrahydroisoquinoline. Ai. J. 393 – 402. S. Dispos. [23] D. Baxter. R. Dispos.3. 11. K. Dispos. Vidgren. R. Takahashi. in rats and dogs. Chem. J. 2001. T. Pharmacol. Catechol-O-methyltransferase-catalyzed rapid O-methylation of mutagenic flavonoids. Zhu. C. M. 164 – 172. S. Biol. Ziegler. D. 1994.S. Rash. L. 178 – 184. P. U. Zhu. P. A. 4202 – 4210. Rogan. 69. Szumlanski. J. B. Kinetics of human soluble and membrane-bound catechol O-methyltransferase: a revised mechanism and description of the thermolabile variant of the enzyme. S.-M. Tilgmann. Kohli. T.

E. Rev. Phenotyping and genotyping studies of thiopurine S-methyltransferase in Kazaks. Rinehart. Drug Metab. Wrighton. V. A. C. and humans. K. A. Pharmacol. Weinshilboum. T. E. Ring. Lennard. [40] H. its metabolism and an overview of its biological effects. 1999. 1 – 15. Iyer. Dispos. Evans. Y. Bonacorsi Jr. Mass. Species differences in the N-methylation and quaternization of [14C]pyridine. Heim. a sulfhydryl-containing vasopeptidase inhibitor in humans. S. 961 – 970. Eckstein. R. A. N. Kronbach. A. A. 1762 – 1766. . Aposhian. Res. B. Toxicol. Kelsell. Brandriff. Gillespie. Malhotra. [34] M. Pharmacogenetics as a molecular basis for individualized drug therapy: the thiopurine S-methyltransferase paradigm. J. DSouza. K. Roongta. Weerakkody. Rinehart. 1063 – 1069. 2001. Ferroni. A major human arsenic metabolite. Part A 2004. R. Drug Metab. P. Wieben. J. Toxicon 2004. M. Remarkable substrate-inhibitor properties of nicotine enantiomers towards a guinea pig lung aromatic azaheterocycle N-methyltransferase. J. E. Cassida. K. Brooks. Verburg. [35] R. [36] J. Res. C. J. 600 – 607. Weinshilboum. S-Methylation as a bioactivation mechanism for mono. Qu. R. M. Kligerman.CHEMISTRY & BIODIVERSITY – Vol. Sakurai. V. 16. Yildiz. 2003. Drug Metab. J. 22. [37] H. 2001. Goldberg. Kripalani. R. Comparative biotransformation of radiolabeled [14C]omapatrilat and stable-labeled [13C2 ]omapatrilat after oral administration to rats. L. 855 – 862. Nicotine. Smith. Andrewes. Biophys. E. N. Jacob III. 397 – 419. Sonawane. 79 – 115. S. 67. Vaccharajani. Drug Metab. B. Thiopurine methyltransferase pharmacogenetics: human gene cloning and characterization of a common polymorphism. Ngo. J. 1996. Quistad. 2002. Methylated trivalent arsenic species are genotoxic. 629 – 637. M. Mitroka. Drug Metab. 2001. Drug Metab. J. K. H. Drug Metab. B. R. Wu. J. D. D. Rehmel. J. Thiopurine pharmacogenetics: clinical and molecular studies of thiopurine methyltransferase. M. Enzymatic methylation of arsenic species and other new approaches to arsenic toxicity. K. Zhou. Toxicol. Wessel. Mosca. B. D. Huang. A. S. Ruterbories. N. H. Krynetski. 29.. R. Cundy. K. 60 – 69. 1985. 128. in humans. Biochem. Commun. 2005. Benowitz. 1997. Thomas. Rash. G. 877 – 882. Peter. a novel thienopyridine. A. Waller. Arsenic toxicology: five questions. M. New metabolic pathways of a-lipoic acid. 17 – 30. [38] E. D. A. Ye Tian. Dimethylarsine and trimethylarsine are potent genotoxins in vitro. J. Kitchin. Tennant. Kurihara. a thienopyridine antiplatelet agent. T. 14. Roop. [41] M. B. F. Smith. dimethylarsinic acid. N. Li. L. Wait. T. Res. 151 – 157. Res. 8. Waller. Health. Toxicol. Aposhian. Bonacorsi Jr. Dispos. [39] R. T. Kurihara. Chem. Mitroka. Res. Pacifici. E. Hermann. Otterness. W. 355 – 361. K. Chem. Annu. Godin. 2006. J. 10. Toxicol. Ginsberg.and dithiocarbamate pesticides as aldehyde dehydrogenase inhibitors. Cullen. T. A. Chem. A. Dispos. B. Farid. 2006. S. S. L. A. A. Hukkanen. A. A. A. M. Stereoselective metabolism of prasugrel in humans using a novel chiral liquid chromatography-tandem mass spectrometry method. C. V. M. C. Chem. E. Khan. S. 5 (2008) 2309 [29] G. A. N. Kurihara. Russ. Bonacorsi Jr. Res. Lee. Giulianotti. Blair. S. G. J. 19. 1995. with the cytochromes P450. J. 601 – 605. S. 57. Wallace.. Dispos. Wickremsinhe. 2006. The disposition of prasugrel. Environ. Toxicol. Drug Metab. Her. 619 – 632. J. 26. Toxicol. C. C. 34. R. P. L. Engel. Pietrabissa. requires reduced glutathione to induce apoptosis. 994 – 1003. 2007. S. Physiologically based pharmacokinetic (PBPK) modeling of caffeine and theophylline in neonates and adults: implications for assessing childrens risks from environmental agents. A. Hattis. A. [31] K. 34. Schupke. P. Szumlanski. D. 2007. Interactions of two major metabolites of prasugrel. P. Khan. Wei. D. 16. W. Toxicol. 35. H. J. Res. 342 – 349. 2001. M. Malhotra. C. R. B. Dispos. F. [30] J. S. Aposhian. R. Kasper. Gomeni. 29. B. M. K. Dispos. Spurr. E. E. Metabolism of [14C]omapatrilat. J. S. Styblo. Rinehart. Xenobiotica 1996. Rev. Xenobiotica 1988. Malhotra. Captopril methylation in human liver and kidney: interindividual variability. [32] J. W. Aronov. J. Dispos. Dispos. 37. C. 43. R. G. L. P. Hempel. J. T. M. G. Iyer. 35. M. S. Pharmacol. Staub. dogs. 15. Farid. 31. and humans. [33] C. Chem. V. 2003. Kripalani. 2005. P. Pharm. 917 – 921. dogs. A. Sparks. C. 29. B. D. J. N. C. R. Waalkes. Res. Jemal. Farid. R. J. J.. Pharm. Iyer. 15. T. DNA Cell Biol. 312 – 316. M. Metabolism of [14C]gemopatrilat after oral administration to rats. 67 – 75. 1096 – 1104. Zhang. A. Metabolism and disposition of nicotine. Patterson. Caldwell. K. 297 – 329. Sakurai. J.

S. K. Toxicol. Metabolism of arsenic in primary cultures of human and rat hepatocytes. Chem. 16. 1051 – 1057. P. Schaid. 172 – 178. Feldmann. Iwata. Res. Aposhian. C. Res. D. S. Wieben. 12. H. M. Y. 616 – 624. Suzuki. L. 2000. 541 – 575. and omega human glutathione transferase. M. J. M. Del Razo. . Toxicol. 1995. Biochem. Hirano. Klumpp. Toxicol. Enzymatic methylation of arsenic compounds: assay. G. V. 2005. R. Sulfonation in Pharmacology and Toxicology. DNA damage induced by methylated trivalent arsenicals is mediated by reactive oxygen species. Toxicol. Toxicol. 2003. P. Morgan. R. C. 2006.-Biol. R. O. Salavagionne. T. Aposhian. Kala. Purine nucleoside phosphorylase in the presence of dihydrolipoic acid is a route for reduction of arsenate to arsenite in mammalian systems. India. Chowdhury. E. 1627 – 1634. Kopplin. Chem. R. R. 2004. A. E. A. U. Res. Zakharyan. D. Zakharyan. 1993. Wood. M. p. C. F. L. K. 371 – 378. R. Zakharyan. V. G. A. H. T. A. Mukherjee. 13. V. A chemical hypothesis for arsenic methylation in mammals. F. 14. Obach. S. 14. Prater. Res. 2003. Gonzalez-Ramirez. 18. C. H. Lee. 1999. M. Y. Aposhian. R. B. K. 16. 1995. R. Maiorino. C. B. A. Res. Zakharyan. Res. G. 243 – 249. W. Board. Toxicol. Silbergeld. A. Wu. 823 – 843. H. Chem. D. Hamilton. 88. Identification of dimethylarsinous and monomethylarsonous acids in human urine of the arsenicaffected areas in West Bengal. Toxicol. 159 – 176. S. M. G. W. Suzuki. 692 – 698. A. Arsenic metabolism and thioarsenicals in hamsters and rats. glutathione. M. Aposhian. Cebrian. Chem. R. Res. Toxicol. Gurzau. Lu. D. Bogdan. K. E. 2004. Res. Sartorelli. Yip. Res. Kauffman. Eds. C. Jaspars. 1624 – 1631. L. Suzuki. B. Biological activity of inorganic arsenic and antimony reflects oxidation state in cultured human keratinocytes. New York. 58. 17. M. B. J. U. 2002. Chem. B. A. Tsaprailis. Y. Arsenate reductase II. Sulfur-containing arsenical mistaken for dimethylarsinous acid [DMA(III)] and identified as a natural metabolite in urine: major implications for studies on arsenic metabolism and toxicity. Weinshilboum. C. Pharmacol. 5 (2008) [42] [43] [44] [45] [46] [47] [48] [49] [50] [51] [52] Reznikova. W. Sampayao-Reyes. H. Mason. M. S. 1029 – 1038. Chem. E. Styblo. Kala. A. Green. Healy. Drug Metab. Thomas. Radabaugh. Toxicol. D. S. Cullen. Appl. Tircus. N. A. 560 – 565. Biol. Formation and urinary excretion of arsenic triglutathione and methylarsenic diglutathione. 693 – 697. E. 2007. Liebler. Weinshilboum. R. LeCluyse. L. R.  Human monomethylarsonic acid (MMAV ) reductase is a member of the glutathione-Stransferase superfamily. Fisher. Chem. K. A. Yamazoe. D. Raftogianis. A. Zakharyan. I. 2006. A. C. H. Fisher. F. and properties of arsenite methyltransferase and monomethylarsonic acid methyltransferase of rabbit liver. Annu. H. Wang. Chem. Tsaprailis. 8. Rev. Yamauchi. Toxicol. J. K. Aposhian. R. 89 – 114. Chem. W. Chem. M. Chem. Naranmandura. J. Pharmacogenetics of sulfotransferases. R. H. L. W. Toxicol. Dart. 1086 – 1091. A. Lambert. D. D. T. 15. Lee. partial purification. 19. 135. 2003. Aposhian. Avram. B. G. 1010 – 1018. Rev. M. Res. Healy. Rice. M. R. Interact. 2000. Chem. R. M. R. Chowdhury.  Human arsenic methyltransferase (AS3MT) pharmacogenetics: gene resequencing and functional genomics studies. D. H. K. Coughtrie. C. Lieberman. Polymorphism in the human monomethylarsonic acid (MMAV ) reductase/hGST01 gene and changes in urinary arsenic profiles. Ogra. X. Styblo. 1287 – 1295. J. Nesnow. Aposhian.2310 CHEMISTRY & BIODIVERSITY – Vol. Hansen. M. M. 2001. Chem. K. V. M. Wang. Eckloff. T. V. Dekker. 40. Res. Pharmacol. T. S. Res. M. Zakharyan. L. J. 1507 – 1513. G. Thompson. B. Naranmandura. T. Thomas. Suzuki. Avram. P. J. Chem. Nagata. H. M. Garcia-Vargas. M. Suzuki. Wood. G. Kadiiska. V. R. Interactions of sodium selenite. 36. E. 281. Toxicol. H. B. G. Res. 2002. L. arsenic species. Res. M.  Human phenol sulfotransferases SULT1A2 and SULT1A1. M. 1999. Marnell. N. L. Raah. Chem. 20. Blanchard. 7364 – 7373. Toxicol. Toxicol. X. Mandal. Toxicol. Ma. 15. Thomae. H. Gurzau. Comparative in vitro methylation of trivalent and pentavalent arsenicals. Chem. 2001. Pharmacol. Toxicol. Occurrence of monomethylarsonous acid in urine of humans exposed to inorganic arsenic. J. Mass. 605 – 616. D. K. V. Hernandez. G. Walsh. 17. M. Trivalent arsenicals are bound to proteins during reductive methylation. SampayoReyes. The role of sulfotransferases (SULTs) and UDP-glucuronosyltransferases (UGTs) in human drug clearance and bioactivation. in Drug Metabolizing Enzymes: Cytochrome P450 and Other Enzymes in Drug Discovery and Development. 2004. J. B. G. Roop. R. B. R. M. Milne. Cullen. G. C. A.

Sundaram. Yli-Kauhaluoma. R. W. Wong. Graphics 2002. J. 2089 – 2097. Leyh. SULT1A1. Petrotchenko. J.  Histidine residues in human phenol sulfotransferases. Campbell. U. Hauser. Boeing. Liu. E. Biochem. Comparison of 2-aminophenol and 4-nitrophenol as in vivo probe substrates for the major human hepatic sulfotransferase. N. characteristics. J. Cheng. 1991. Bertozzi. Teubner. D. R. 2007. Aust. VEGA: a versatile program to convert. 27 – 40. N. J.CHEMISTRY & BIODIVERSITY – Vol. W. G. Pomplun. 14. H. H. 5 (2008) 2311 [53] [54] [55] [56] [57] [58] [59] [60] [61] [62] [63] [64] [65] R. Alakurtti. Kulkarni. Ther. I. He. R. B. D. Biophys. Ma. Jaeger. M. 310. Tabrett.povray. Biochem. Pharmacol. W. Variable sulfation of dietary polyphenols by recombinant human sulfotransferase (SULT) 1A1 genetic variants and SULT1E1. Coughtrie. 36. Runge-Morris. A proposed nomenclature system for the cytosolic sulfotransferase (SULT) superfamily. McCann. I. Regulation of sulfotransferases by xenobiotic receptors. 27325 – 27330. 352 – 358. Chem. 299 – 307. Zhang. R. 280. W. Reznick. M.  Human and rat liver phenol sulfotransferase: structure-activity relationships for phenolic substrates. C. Dispos. 2003. POVRay. Drug Metab. 2006. W. Exp. Bloomer. Kuhlow. J. L. Welshons. J. Weinshilboum. A. Pharmacol. R. Curr. Opin. interindividual variation and re-evaluation of the suitability of 4-nitrophenol as a substrate probe. 1474 – 1481. C. 273. Chen. J. T. W. Miksits. H. Weinshilboum. Compendium of gene expression profiles comprizing a baseline model of the human liver drug metabolism transcriptome. The physical biochemistry and molecular genetics of sulfate activation. Toxicol. The sulfuryl transfer mechanism. Pedretti. McManus. Crit. J. Pharmacol. Mol. Biochem. 6. L. 41482 – 41486. T. G. E. R. A. Xenobiotica 2007. VMD. G. Biol. Biochem. Y. T. X. J. Reprod. 63 – 69. S. 2001. 24. 813 – 819. Duggleby. J. L. Pharmacogenetics 2004. Castle. Maier-Salamon. handle and visualize molecular structure on windows-based PCs. 1998. H. Dispos. J. Biol. Chen. Pedersen. Apak. 67. 139 – 177. M. Maiti. Robertson. tetrabromobisphenol A. Engelke. N. S. Vistoli. Szekeres. Pharmacol. 1993. Interactions between dietary chemicals and human sulfotransferases – Molecular mechanisms and clinical significance. H. Martin. S. 29. Y. D. B. Rev. 199 – 211. 325 – 331. H. W. Hansch. A. R. www. Z. C. O. V. Buck. V. Drug Discovery Dev. Swedmark. 740 – 746. Negishi. Toxicol. K. 693 – 708. 504 – 510. G. demonstrates improved selectivity with 2aminophenol.  Human cytosolic sulphotransferases: genetics. Rushmore. E. Drug Metab. Sit. M. 32. C. Chem. E. Richards. R. S. Villa. Regioselective sulfonation of dopamine by SULT1A3 in vitro provides a molecular explanation for the preponderance of dopamine-3-O-sulfate in human blood circulation. Freimuth. Riches. Biochem. Biochem. G. Nakagawa. N. Phenol sulfotransferase 1A1 activity in human liver: kinetic properties. M. Slatter. M. Pharmacol. C. 2004. O. H. Taskinen. 74. H. Pabel. C. T. Duffel. Y. Coughtrie. S. Kocarek. Ishii. H. S. Kakuta. L. Biochem. 275. Y. 2007. Res. L. 654 – 658. toxicological aspects. M. U. M. T. 19. Tsoi. Lehmler. I. J. Ung. Johnston. K. Haslinger. A. H. K. Glatt. 1994. 2001. A. Kunert. Redox regulation of human estrogen sulfotransferase (hSULT1E1). 2007. Depletion of hepatic 3’-phosphoadenosine 5’-phosphosulfate (PAPS) and sulfate in rats by xenobiotics that are sulfated. T. Pharmacol. Kim.  Human exposure to bisphenol A (BPA). 35. Suzuki. The structure of human SULT1A1 crystallized with estradiol. 2005. Biol. C. Olea. 74. J. 515 – 542. and its analogues in rat hepatocytes. R.and tricyclic phenols by rat liver aryl sulfotransferase isozymes. 482. Sulfation of resveratrol in human liver: evidence of a major role for the . 2007. Templeton. Nagar. 1995. Parker. Mol. S. E. J. Cho. Gamage. C. 275 – 285.-J. 3. A. 73. Coughtrie. C. 2005. 522 – 528. W. Xenobiotica 2006. Drug Metab. Caguyong. M. M. Mangold. 502 – 515. W. Pharmacol. C. 41. Y. Biotransformation and cytotoxicity of a brominated flame retardant. Van Loon. M. Sulfation of di. P. Curr. Curr. J.uiuc. K. Arch. 21. H. 66. A. L. G. Dai. A. Chem. 37. H. J. Res. 1987. Kosiainen. J. Pharmacol.unimi. 938 – 962. Tsvetanov. J. Ames. 2000. 6. Mutat. Sulfotransferases as targets for therapeutic intervention. 2007. R. M.  Hydroxylated polychlorinated biphenyls are substrates and inhibitors of human hydroxysteroid sulfotransferase SULT2A1. L. 1355 – 1361. Klaasen. Ulrich. Biochem. M. Meinl. Coughtrie. 47 – 49 (www. H. Itä M. Khoo. Drug Metab. www. S. E.ddl. Armstrong. D. Ogata. J. M. 1420 – 1425. Mol. Vandenberg. Biosynthesis of PAPS in vitro by human liver. 28. 2005. G. Thalhammer. W. Sulfation in dogs.ks. Marcus. M. T.

Pharmacol. P. 1985. 651 – 657. G. Res. 19. Chem. Current status of the cytosolic sulfotransferases in the metabolic activation of promutagens and procarcinogens. Res. T. 24. B. 1994. 423 – 427. Drug Metab. Interact. Johnson. L. 2000. 1994.-Biol. A. W. 87 – 105. Negishi. G. K. Y. 329 – 341. Res. Toxicol.-Biol. Dispos.-Biol. Dispos. Duffel. Ogura. D. 800 – 803. Y. Roles of electrophilic sulfuric ester metabolites in mutagenesis and carcinogenesis by some polynuclear aromatic hydrocarbons. N. Dispos. Banoglu. E. O. Walle.5-dichlorophenyl)succinimide in rats: evidence for bioactivation through alcohol-O-glucuronidation and O-sulfation. Metabolism and excretion of trovafloxacin. 25. Drug Metab. J. Influence of substrate structure on the catalytic efficiency of hydroxysteroid sulfotransferase STa in the sulfation of alcohols. A. Role of enterohepatic circulation. Dalvie. 1231 – 1240. Pedersen. Res. J. Carrupt. 1994. Isomer. Kakuta. in rats and dogs. D. Interact. 35. Hirschelman. developmental aspects and sex differences. 1304 – 1310. and mouse metabolism of resveratrol. Pezzuto. Navetta. Meisheri. 33 – 45.-J. Falany. 92. Drug Metab. Testa. A. 92. W. Enzymatic and nonenzymatic sulfation mechanisms in the biological actions of minoxidil. Banoglu. minoxidil analogs and catecholamines. Res. Lewis. Puddington. Y. Chem. V. G. 30. Watabe. Chism. Dispos. Chen. Rickert. King. van Breemen. Interact. K. Chem. Excretion and metabolism of trovafloxacin in humans. G. Sharma. D. A. Bamforth. 8. Toxicol. G. Otake. Duffel. R. Okuda. E. 278 – 285. H. a new quinolone antibiotic. 1251 – 1258. S. A. N. Toxicol. Phase 2 metabolites of N-hydroxylated amidines (amidoximes): synthesis. R. 164). M.and sex-specific bioactivation of mononitrotoluenes. P. G. 2000. 18. Shin. Meerman. H. H. Curr. b) A. Lee. Khosla. Toxicol. K. A. Yu. H. 13. Molecular cloning and functions of rat liver hydroxysteroid sulfostranferases catalysing covalent binding of carcinogenic arylmethanols to DNA. 1999. A. Chem. E. 5. E. 319 – 326. in vitro formation by pig hepatocytes. K. Walle. 9. M. 73 – 76 (correction 1995. 10. Interact. Toxicol. Christiansen. Kosmeder. a) J. M. C. 2002. Walle. Structure-function modeling of the interactions of N-alkyl-N-hydroxyanilines with rat hepatic aryl sulfotransferase IV. 1994. J. Xenobiotica 2000. D. S. Miller. S. H. 12. Banoglu. Caldwell. B. K. Harvison. E. Miller. Y. 67 – 74. Wilson. W. J. R. Interact. K. Duffel. 271 – 279. rat. 13. M. E. M. Dalvie. Walle. Brighty. Chem. 1. J. Hiratsuka. Satsukawa. Pharm. Chem. C. P. 1996. 602 – 609. Aberg. Coughtrie. K. Clement.-Biol. Rankin. Gilissen. Khosla. N. Chirality 1993. Res. A. J. Vincent. Chem. J. N. 5 (2008) [66] [67] [68] [69] [70] [71] [72] [73] [74] [75] [76] [77] [78] sulfotransferases SULT1A1 and SULT1E1. Chem. Enantioselective sulfation of b2-receptor agonists by the human intestine and the recombinant M-form phenolsulfotransferase. U. 321 – 328. Toxicol. 2005. Singer. Duffel. Xenobiotica 2005. 92. H. Drug Metab. 351 – 362. T. B. Metabolism of the nephrotoxicant N-(3. T. Sulphonation of Nhydroxy-2-acetylaminofluorene by human dehydroepiandrosterone sulphotransferase. 1997. The same enzymes catalyze sulfation of minoxidil. Pesola. Y. D. Y. Surh. Hartman. R.  Human. Chow. 1993. M. 1907 – 1914. and platelet activity. 253 – 261. L. 14.2312 CHEMISTRY & BIODIVERSITY – Vol. Girreser. A. intestinal. Chem. Sulfonation in chemical carcinogenesis – history and present status. Structure-genotoxicity relationships of allylbenzenes and propenylbenzenes: A quantum-chemical study. P. M. Stereoselective sulfate conjugation of isoproterenol in humans: comparison of hepatic. 991 – 1003. Mehta. Tsai. M. Sulfation of carcinogenic aromatic hydroxylamines and hydroxamic acids by rat and human sulfotransferases: substrate specificity. 1101 – 1119. Wilson. A. Drug Metab. Ringer. 1994. W. S. 2001. Studies on the interactions of chiral secondary alcohols with rat hydroxysteroid sulfotransferase STa. 1 – 30. T. J. 92. Res. Cui. J. 45. Biochem. Chem. K. 1994. M. and mutagenicity testing. 25. Li. D. U. A. 7. R. W. W. 92. P. G. K. R. Importance of peri-interactions on the stereospecificity of rat hydroxysteroid sulfotransferase STa with 1-arylethanols. Chirality 1998. 1997. C. Banoglu. W. J. . J.-Biol. S. Chem. 1996.

Xenobiotica 1986. J. 691 – 696. J. 81. Burchell. Chowdhury. J. Pharmacol. 2004. Vertebrate UDP-glucuronosyltransferases: functional and evolutionary aspects. ´ Belanger. 72. R. Shiraga. Ikushiro. Biochem. D. Pharmacogenomics J. A. Guillemette. G. K. K. Cheng. Biochem. Pagliara. C. The UDP glycosyltransferase gene superfamily: recommended nomenclature update based on evolutionary divergence. 121 – 133. Noguchi. Albert. Mackenzie. 17 – 26. M.CHEMISTRY & BIODIVERSITY – Vol. S-Sulphoconjugation of alicyclic. Wong. S. K. Louisot. D. C. Peters. 239 – 243. Little. R. D. K. chemical proof. Welzenbach. 347 – 369. Y. Lancet. Gaillard. Permert. [90] P.flinders. Chen. N. W. P. B. J. 2004. R. N-Sulphation of desipramine in the rat brain. K. P. J. Toxicol. T. [80] K. Johansson. Ishii. Hum. K. [81] J. Z. Annu. Y. M. 581 – 616. W. Schachter. Knecht. Fournel-Gigleux. 3385 – 3400. I. P. 5 (2008) 2313 [79] T. Zecri. D. 68. Glucuronidation of drugs – A re-evaluation of the pharmacological significance of the conjugates and modulating factors. Identification. P. 677 – 685. W. Karlsson. I. 29. Tephly. J. Ther. Chowdhury. Pharmacol. Biochem. 56. 32. 3. Iyanagi. Pharmacogenomics of human UDP-glucuronosyltransferase enzymes. E. 262. Parker. Tada. Dickinson. H. Owens. F. M. Cooke. Krajewski. 136 – 158. Xenobiotica 1999. C. Glucuronidation and the UDP-glucuronosyltransferases in health and disease. a novel DNA methylation inhibitor. Chowdhury. Matthews. M. W. Müller-Hartwieg. Annu. 263 – 272. [84] R.-J. UDP-Glucuronosyltransferases. Ther. A. Lynx. Carrupt. [86] Y. T. U. Ther. O. Pharmacol. Carrupt. Pharmacol. Wong. J. O. Klotz. E. ´ Streiff. 32. Iwasaki. 2005. P. 292 – 310. Ramaswamy. Pharmacol. 23. Tipton. Pharmacol. 168 – 176. alkyland aryl-amines in vivo and in vitro. D. K. M. 1992. C. 1987. Jakoby. Pharmacol. P. D. Amine N-sulfotransferase. Deville-Bonne. Drug glucuronidation in humans. Toxicol. Pagliara.-C. K. Hinterding. F. K. 40. Mackenzie. Agbaria. W. 1998. H. 51. R. 1989. Anabolism of amdoxovir: phosphorylation of dioxolane guanosine and its 5’phosphates by mammalian phosphotransferases. H. C. Burka. K. Krishnan. and biological characterization of the biologically active species and its enantiomers. V. 7. Wagner. S. A. Bratton. Guillemette. Wells. Ritter. V. G. expression. [88] B. Guillemette. Biol. Ben-Zvi. S. S. 2003. 25 – 49. Pharmacokinet. P. J. Green. Chem. Caron. and disease. Hata. Burchell. 26. B. J. 10039 – 10043. N. Noda. H. P. Y. Ben-Kasus. J. T. Biochem. Drug Metab. Novel immunomodulator FTY720 is phosphorylated in rats and humans to form a single stereoisomer. Tuckey. Veron. Yamazoe. K. Kessler. B. 341 – 347. Borroto-Esoda. Rev. K. A. Iyanagi. [85] H. R. T. Clin. J. L. J. 1996. R. A. [82] C. 2003. B. Testa. Burchell. J. McKee. Rev. Bock. A. Pharmacogenet. T. J. Testa. O.-I. Y. Marquez. Pharmacol. Gambertoglio. Kroemer. G. 651 – 659. Xenobiotica 1996. Metabolic activation of zebularine. M. W. Kelley. Radominska-Pandya. Iwasaki. Guerini. 48. B. Bairoch. 5373 – 5377. Med. 2000. Strassburg. M. Hansen. McKenzie. 3’-Azido-3’-deoxythymidine (AZT) is a competitive inhibitor of thymidine phosphorylation in isolated rat heart and liver mitochondria. Glucuronidation and its role in regulation of biological activity of drugs. 255 – 269. Bock. Pharmacogenetics 1997. A. 281 – 290. Feng. E. Biochem. 2005. Francotte. R. H. Acta 1998. 66. W. K. 30. M. M. A. Dispos. M. Chem. Xenobiotica 2000. S. Mackenzie. P. Nomenclature update for the mammalian UDP-glycosyltransferase (UGT) gene superfamily. Ritter. Miners. T. J. Y. G. H. P. P. Clin. P. Nebert. 2006. Kim. 16. L. Miners. Giroud. Rev. Gregory. Pharmacol. N-Sulphoconjugation of amines by human cytosolic hydroxysteroid sulphotransferase. 1991. W. Sanders. 1992. 60. 15. Ohno. Zidovudine phosphorylation after short-term and long-term therapy with zidovudine in patients infected with the human immunodeficiency virus. M. A. K. Zhu. Phosphorylation of anticancer nucleotide analogs by human mitochondrial deoxyguanosine kinase. 1879 – 1888. Chim. I. C. A historical overview of the heterologous expression of mammalian UDP- . Zollinger. 330 – 341. E. Metabolism and disposition of luminol in the rat. Helv. Intrinsic and intramolecular lipophilicity effects in O-glucuronides. I. [89] http://som. 1035 – 1040. Simeon. Coughtrie. F. Mulder. Nebert. Bock. G.  Human UDP-glucuronosyltransferases: metabolism. W. P. Lipophilicity profiles of ampholytes. Owens. 70. J. Magdalou. Brinkmann. 261 – 289. J. Genomics 2005. [87] A. K. I. 43. S. A. 97. 1997. Shiraga. B. T. Chem. Pharmacol. [83] K. T. in human bladder carcinoma cells. B. K.

2007 – 2015. R. Res. P. H. J. . Furniss. Mori. R. Tian. McKinnon. Mol. M. P. S. J. 2006. M. Pharmacol. P. Miley. A. O. F. P. Y. 1463 – 1473. J. Knights. Sorich. Drug Metab. R. O. P. 2003. H. Tracy. 65. Strassburg. Green. J. Pharmacol. Smith. 405 – 414. Wrighton. J. Iwai. Toxicol. J. A. Takeuchi. Y. 1617 – 1626. Czernik. T. Glucuronidation of trans-resveratrol by human liver and intestinal microsomes and UGT isoforms. Iyer. A. P. 2001. 53 – 56. Zhang. Puig. 2006. B. A. P. Structural and functional studies of UDP-glucuronosyltransferases. Battaglia. Xenobiotica 1988. M. Mackenzie. L. 1201 – 1210. McKinnon. Tuckey. Mackenzie. 2005. 1531 – 1539. R. Sorich. Comparative modelling of the human UDP-glucuronosyltransferases: insights into structure and mechanism. A. I. Miners. Pharm. Polymorphism of UDP-glucuronosyltransferases and drug disposition. J. 18. Paine. 58. M. Radominska-Pandya. M. Drug glucuronidation by human renal UDP-glucuronosyltransferases. T. Ethell. I. Drug Metab. P. Amino terminal domains of human UDP-glucuronosyltransferases (UGT) 2B7 and 2B15 associated with substrate selectivity and autoactivation. Lewinsky. The role of hepatic and extrahepatic UDP-glucuronosyltransferases in human drug metabolism. Drug glucosidation. Biochem. 2006. Fisher. Locuson. O. C. Genetic variants of human UGT1A3: functional characterization and frequency distribution in a Chinese Han population. S. Lim A. Puhlavisto. Pharmacol. 22. Chen. 46. X. Pharmacol. P. Taskinen. C. 6. Conjugation of catechols by recombinant human sulfotransferases. J. 1187 – 1197. Y. Sorich. Burchell. 44. Ou. Green. Pharmacol. 1990. M. S. Bradford. C. Zuo. Mol. B. W. Biochem. Dispos. W. S. J. M. Brierley. Rev. Burchell. 33. Pharm. M. M. 91 – 99. M. L. 6. Drug Metab. Mol. Miners. Med. 81 – 89. M. Genetic multiplicity of the human UDP-glucuronosyltransferases and regulation in the gastrointestinal tract. M. M. H. I. P. 155 – 168. Dispos. J. Smith. Xenobiotic nuclear receptor-mediated regulation of UDP-glucuronosyltransferases. Zeng. 289 – 298. T. C. L. J. M. R. Curr. Irshaid. Rapid prediction of chemical metabolism by human UDPglucuronosyltransferase isoforms using quantum-chemical descriptors derived with the electronegativity equalization method. G. T. 37. Activity and expression of various isoforms of UDP-glucuronosyltransferases are differently regulated during hepatic regeneration in rats. Rev. P. 301 – 308. Miners. Glucuronidation of bioflavonoids by human UGT1A10: structure-function relationships. Curr. O. W. Pharmacol. A. 273 – 297. Smith. McGurk. Green. Radominska-Pandya. G. Drug Metab. 46. Xenobiotica 2005. Hood. Chem. K. Z. Venkataramanan. R. 5311 – 5317. C. Annu. O. 117 – 129. and soluble catechol Omethyltransferase: structure-conjugation relationships and predictive models. Crystal structure of the cofactor-binding domain of the human phase II drug-metabolism enzyme UDP-glucuronosyltransferase 2B7. 817 – 899. E. Multiple pharmacophores for the investigation of human UDP-glucuronosyltransferase isoform substrate selectivity. Strelevitz. In vitro-in vivo correlation for drug and other compounds eliminated by glucuronidation in humans: pitfalls and promises. I. 31. X. A. 73. Res. V. Furimsky. W. M. 2005. Kapetanovic. Y. Ther. Predicting human drug glucuronidation parameters: application of in vitro and in silico modeling approaches. 47. 2006. Xenobiotica 2007. J. J. Rev. Li. 1 – 25. 2005. McKinnon. Med. B. Tang. Strom. K. J. Houston. P. Zhou. D. Drug Metab. R. 2004. Little. 2005. Chen. Chem. A. P. H. M. B. Sato. 498 – 511. E. Miners. J. A. J. Mackenzie. 2007. 34. J. Drug Metab. A. J. W. M. C. Mackenzie. Xie. Wang. 141 – 160. Lin. A. 369. J. S. B. Redinbo. Smith. Coughtrie. I. N. 71. Endogenous substrates for UDP-glucuronosyltransferases. Elliot. Lewis. Involvement of UGT-glucuronosyltransferases in the extensive liver and intestinal first-pass metabolism of flavonoids. S. J. Z. 55. Smith. Miners. M. Zielinska. 31. O. Pharmacol. 1005 – 1012. 2004. B. J. Miners. Drug Metab. McKinnon.-K. 469 – 479. Maruo. Mackenzie. 2007. G. Burchell. 5 (2008) [91] [92] [93] [94] [95] [96] [97] [98] [99] [100] [101] glucuronosyltransferase isoforms over the last twenty years. H. J. 2003. UDP-glucuronosyltransferases. Bhasker. A. Zhang. Winkler. S. A. J. Curr. A. H. J. A. Y. 1462 – 1467. Brill. 1999. M. Keenan. Zuo. K. Tephly. 35. Zhang. R. 59. Pharmacol. D. 24. S. A. J. Pharmacophore and QSAR modeling: complementary approaches for the rationalization and prediction of UDP-glucuronosyltransferase 1A4 substrate selectivity. E. J. Bratton. Biol. J. R. 2004.2314 CHEMISTRY & BIODIVERSITY – Vol. A. 2001. B. Biochem. I. Ho. Pharm. C. Sorich. 6. B. A. 1998.

2006. T. Quantitative regioselectivity of glucuronidation of quercetin by recombinant UDP-glucuronosyltransferases 1A9 and 1A3 using enzymatic kinetic parameters. M. 857 – 864. R. Bauman. Wong. W.  Human UDPglucuronosyltransferase. M. J. van Iersel. Mol. 4. Springer. C. V. Tam. Kostrubsky. X. oestradiol and morphine. Regioselectivity of phase II metabolism of luteolin and quercetin by UDP-glucuronosyltransferases. A. J. Rietjens. J. M. Pharmaceutics 2007. K. Vervoort. Iwabuchi. 1A9 and 2B15. M. T. 943 – 954. I. Clarke. 2003. P. Alton. Hu. Patten. S. 34. D. 283 – 288. 1520 – 1530. UGT1A8. Samata.  Hydrolysis in Drug and Prodrug Metabolism – Chemistry. Xenobiotica 2006. P. 1998. 322. M. M. Grassman. 998 – 1006. S. 131 – 160. Potential implications in acetaminopheninduced hepatotoxicity. R. Drug Metab. 31.and 6-glucuronide metabolites in humans and animals. Green. R. Toxicol. 28. M. T. van Bladeren. J. J. Testa. H. Pharmacokinet. B. T. Tephly. 314 – 320. Vree. Vervoort. 282 – 293. van der Woude. Mutlib. 701 – 709. Zheng. li. L. glucuronidates dihydrotestosterone to a monoglucuronide and further to a structurally novel diglucuronide. Berlin. J. R. 2006. C. A. M. J. N. Styczynski. M. 2003. M. Chen. 324 – 330. J. J. Boeren. M. Bogaars. Dispos. S. Pharmacol. B. 1995. Novak. L. Burchell. Q. Y. Dispos. and Wiley-VCH. F. Ikeda. Wang. S. Schänzer. Milne. 1102 – 1108. 299. Kulkarni. H. Disposition of flavonoids via enteric recycling: structural effects and lack of correlations between in vitro and in situ metabolic properties. J. Mol. Chen. Pharmaceutics 2007. in Conjugation-Deconjugation Reactions in Drug Metabolism and Toxicity. Somogyi. 2003. Chowdhury. T. 1A6. Wright. Swank. 2002. R. Tam. J. Kauffman. Kurkela. J. G. S. K. R. E. Coffman. X. J. 19. L. Cnubben. Achanfuo-Yeboah. R. R. and Enzymology. M. Liu. S. Dispos. Chen. D. Auyeung. H. C. J. Ikeda. Chem. Miners. M. Ed. Yuan. Drug Metab. 63. L. M. I. Dispos. 5 (2008) 2315 [102] [103] [104] [105] [106] [107] [108] [109] [110] Lin. V. M. T. Pharmacol. M. Enterohepatic cycling and pharmacokinetics of oestradiol in postmenopausal women. Xenobiotica 2005. X. Kostiainen. P. Drug Metab. Mayer. Res. Pharm. Pharmacol. 1837 – 1848. R. Arch. Intestinal and hepatic glucuronidation of flavonoids.. R. 1994.and mixed conjugates and their formation by rat and human phase II in vitro model systems. 2005. 2001. K. L. Rietjens. H. J. Biochemistry. T. K. Finel. J. Tephly. 32. Glucuronidation of anabolic androgenic steroids by recombinant human UDP-glucuronosyltransferases. T. N. Ritter. D. 30. von Moltke. D. 1996. K. D. Toxicol. M. A. M. L. R. T. Toxicol. H. S. J. A. Drug Metab. Oturu. Disposition of flavonoids via enteric recycling: determination of the UGT-glucuronosyltransferase isoforms responsible for the metabolism of flavonoids in intact Caco-2 TC7 cells using siRNA. Greenblatt. I. 17. C. 833 – 845. Biochem. Jia. S. E. Identification of 14 quercetin phase II mono. 873 – 882.CHEMISTRY & BIODIVERSITY – Vol. Court. S. Drug Metab. B. Iannucci. J. Kinetics of acetaminophen glucuronidation by UDPglucuronosyltransferases 1A1. A. Thevis. M. Identification of human UDP-glucuronosyltransferase enzyme(s) responsible for the glucuronidation of ezetimibe (Zetia). The disposition of morphine and its 3. C. M. 2006. Mackenzie. Williams. Christ. 1178 – 1190. Grace Jr. Interindividual variability in acetaminophen glucuronidation by human liver microsomes: identification of relevant acetaminophen UDP-glucuronosyltransferase isoforms. Bai. Genetic regulation of the subcellular localization and expression of glucuronidase. Opioids bind to the amino acids 84 to 118 of UDPglucuronosyltransferase UGT2B7. Boersma. Patrick. Zhen. 345 – 472. 2002. 20. Goosen. Kessler. Dispos. Y. S. S. W. Nation. S. Murai. 460 – 468. R. E. J. M. H. N. H. B. C. P. 662 – 670. F. 1117 – 1124. Goldsmith. D. E. Ghosal. Chem. Cloning and expression of rat liver phenobarbital-inducible UDP-glucuronosyltransferase (2B12) with specificity for monoterpinoids alcohols. S. 2004. Chem. G. B. H. 4. Hapangama. O. Boersma. J. Iwabuchi. Zbaida. Browning. Knosp. Drug Metab. Glucuronidation in the chimpazee (Pan troglodytes): studies with acetaminophen. . 50. Zurich. Exp. 2004. Kuuranne. Duan. Mol. B. Repeated glucuronidation at one hydroxyl group leads to structurally novel diglucuronides of steroid sex hormones. Res. A. and the importance of the metabolites to the pharmacological effects of morphine. X. J. p. 35. 15. Verlag Helvetica Chimica Acta. Jackson. D. 36. T. van der Woude. Drug Metab. Murai. J. Rev. M. Timmer. W. N. 34. M. P. Hu. Biophys. Kessler. Res. Glucuronidation of acetaminophen catalyzed by multiple rat phenol UDPglucuronosyltransferases. Ther. E. Kearney. Weinheim. H.

Uutela. G. L. D. U. Hao. J. Properties of acyl glucuronides: implications for studies of the pharmacokinetics and metabolism of acidic drugs. Bouska. B. Stearns. Bernstein. Z. 1257 – 1265. R. D. Predicting the pharmacokinetics of acyl glucuronides and their parent compounds in disease states. Drug Metab. A. Ther. Al-Rohaimi. 34. Stereoselective glucuronidation of zileuton isomers by human hepatic microsomes. L. 58 – 66. Guillemette. P. Dispos. A. A. Bell. J. Locke. P. Grillo. 1999. Dispos. J. Luukkanen. 24. Chem. 356 – 363.2316 CHEMISTRY & BIODIVERSITY – Vol. G. C. J. Nellans. S. T. Stereoselective conjugation of oxazepam by human UDP-glucuronosyltransferases (UGTs): S-oxazepam is glucuronidated by UGT2B15. 10. Drug Metab. 23. Eichelbaum. Xenobiotica 2007. Beattie. Williams. and its metabolites. 2006. Chirality 1993. Ogiso. Springer Verlag. Bekaii-Saab. 7. Park. W. Belanger. 2000. Sweeny. J. Berlin. Harding. 1502 – 1507. K. Finel. Chem. L. Somogyi. M. Eds. B. Riley. Awni. T. R. dog. A. 2002. Journault. P. 1213 – 1249. Dispos. C. Wilson. L. Pharm. D. Faed. Martin. Iwaki. Hum. Toxicol. Stachulski. . W. Bernauer. J. Tephly. J. 1984. Identification of a hydroxylamine glucuronide metabolite of an oral hypoglycemic agent. P. I. 24. its enantiomers. L. J. 1999. Sanders. Maggs. M. Drug Metab. Krishnaswamy. el-Shourbagy. Spahn-Langguth. and human in vitro systems and dog in vivo. M. D. 1995. 9 – 21. A. A. 656 – 665. Sci. 49. Drug Metab. Girard. Curr. D. Green. 310. T. J. V. Acyl glucuronides revisited: is the glucuronidation process a toxification as well as a detoxification mechanism?. N. J. Dispos. L. Howe. Dispos. Dekant. 1996. Studies on the chemical reactivity of 2-phenylproprionic acid 1-Oacyl glucuronide and S-acyl-CoA thioester metabolites. Martin. Opin. Carrupt. Grillo. 28. J. S. W. J. in Stereochemical Aspects of Drug Action and Disposition. Greenblatt. 31. 147 – 163. Dispos. Chen. J. C. Sten. 178 – 185. 5 (2008) [111] [112] [113] [114] [115] [116] [117] [118] [119] [120] [121] Green. Med. Biotransformation of [12C]. Rev. P. Pharmacokinet. Dispos. Drug Metab. Curr. 20. Court. Dube The pharmacokinetics of single oral doses of zileuton 200 to 800 mg. T. Which hydroxy? Evidence for species differences in the regioselectivity of glucuronidation in rat. Tephly. A. UDPGlucuronosyltransferase (UGT) 2B15 pharmacogenetics: UGT2B15 D85Y genotype and gender are major determinants of oxazepam glucuronidation by human liver. M. 149 – 153. J. Acyl glucuronides: biological activity. J. J. D. Drug Metab. Studies on the chemical reactivity of diclofenac acyl glucuronide with glutathione: identification of diclofenac-S-acylglutathione in rat bile. M. Krishnaswamy. S. G. Lewis. 6931 – 6945. Chem. 2006. R. S. 2004. Drug Metab. Kakehi. 497 – 502. P. D. 958 – 964. Drug Metab. Z. Res. Rev. Benet. A. Drug Discovery Dev.and 2-acyl glucuronides of naproxen and their covalent binding to human serum albumin. ´. Testa. L. In vitro regioselective stability of b-1-O. Toxicol. Wong. 37. Miller. B. M. 328 – 329. a) I. H. 2004. Drug Metab. Dispos. 15. D. C. Qvisen. Testa. The so-called interconversion of stereoisomeric drugs: an attempt at clarification. M. Drug Metab. in normal healthy volunteers. 2006. Drug racemization and its significance in pharmaceutical research. M. A. A. R. L. N. Greenblatt. K. P. M. Cavanaugh. 2). Reist. H. Q. 5 – 48. ´ Barbier. Doss. Amberg. A. 3’-Azido-3’deoxythimidine (AZT) is glucuronidated by human UDP-glucuronosyltransferase 2B7 (UGT2B7). J. Identification and characterization of two chloramphenicol glucuronides from the in vitro glucuronidation of chloramphenicol in human liver microsomes. 1992. M. J.4 protein. Prominent but reverse stereoselectivity in propranolol glucuronidation by human UDP-glucuronosyltransferases 1A9 and 1A10. K. 12. 91 – 112. Duan. G. Hua. Drug Metab. S. Liu.and [13C]-tert-amyl methyl ether and tert-amyl alcohol. Cepa. Drug Metab. Kostiainen. 954 – 971. R. Carter. R. M. Sweeny. E. Glucuronidation of amines and hydroxylated xenobiotics and endobiotics catalyzed by expressed human UGT1. M. Lindon. 2007. 1327 – 1336. L. D. S. b) O. Dispos. C. A. C. p. Li. L. B. Machinist. D. Leduc. Res. S. S. C. Gal. 32. 34. 1995. 2006. Knutson. 2003. 1488 – 1494. T. 2002. 29 (Suppl. Carrupt. a) M. B. while R-oxazepam is glucuronidated by UGT2B7 and UGT1A9. Inagawa. von Moltke. R. H. Kerr. I. 1992. 105 – 111. and chemical synthesis. Testa. 1309 – 1317. V. Turgeon. J. Nellans. F. D. H. T. Scheutzow. A. H. B. chemical reactivity. 5. R. Clin. X. 15. Waldon. Exp. J. M. Benet. R. S. Glucuronidation of zileuton (A-64077) by human hepatic microsomes. E. Smith. 30. Pharmacol. R. D. Court. The chemistry and biological activity of acyl glucuronides. von Moltke. 2003. C. Ware. Stachulski. A. H. M. Springthorpe.

S. J. J. X. Glucose autoxidation and protein modification. Lin. 52 – 57. Ojingwa. G. Hashimoto. Chemical and immunological comparison of protein adduct formation of four carboxylate drugs in rat liver and plasma. 2006.  Hydrophilicity/lipophilicity: relevance for the pharmacology and clinical effects of HMG-CoA reductase inhibitors. Shimada. C. Fairchild. I. A. Burlingame. a) M. McDonagh. 1050 – 1057. Azam. Bioactivation of carboxylic acid compounds by UDP-glucuronosyltransferases to DNA-damaging intermediates: role of glycoxidation and oxidative stress in genotoxicity. J. Linetsky. Commun. 659 – 666. Baynes. K. 257. Chem. 30. Hasegawa. Determination of degradation pathways and kinetics of acyl glucuronides by NMR spectroscopy. 2002. H. Life Sci. 6165 – 6176. M. Bauman. b) K. 90. J. B. 2002. Maggs. V. The potential role of autoxidative glycosylation in diabetes. Drug Metab. Reily. S. Rev. Mossine. 2007. Volland. 715 – 725. Acad. 35. Toxicol. Res. 506 – 510. Hosagrahara. reactivity. J. McGurk. Remmel. 243 – 250. Burcham. Y. R. Beaumont. 959. 26 – 37. 1993. Predictability of the covalent binding of acidic drugs in man. B. c) L. T. a new inhibitor of HMG-CoA reductase: human UDP-glucuronosyltransferase enzymes involved in lactonization. Maffei Facino. Burchell. Res. Drug Metab. R.and (S)-ketoprofen. P. J. Mizuma. Lam. Evidence for covalent binding of acyl glucuronides to serum albumin via an imine mechanism as revealed by tandem mass spectrometry. Iwakawa. Y. R. S. B. I. 876 – 886. Wilson. T. S. 2004. J. 12. 245. Walker. Yamada. J. 2007. Sci. 34. G. Benet. H. T. Ding. 20. T. T. Ann. Sci. A. Magdalou. Kohl. S. Superoxide free radical generation by Amadori compounds: the role of acyclic forms and metal ions. Z. Pharm. C. Mayer. T. Mutschler. Xenobiotica 2004. L. c) N. I: Design. Sabramanian. 17. J. a) J. V. Ma. (R). 230 – 236. Ortwert. b) A. K. 9. 25. P. Spahn-Langguth. P. Dispos. J. U. Tranter. Skordi. Pearson. Xenobiotica 2005. Dalle-Donne. Natl. A.CHEMISTRY & BIODIVERSITY – Vol. Interaction of human serum albumin with the electrophilic metabolite 1-O-gemfibrozil-b-d-glucuronide. Amadorins: novel post-Amadori inhibitors of advanced glycation reactions. E. Fournel-Gigleux. 1993. Med. Trends Pharmacol. 24. and tolmetin by 1H-NMR spectroscopy. 55 – 60. The Maillard hypothesis on aging: time to focus on DNA. R. Yoneda. 33. 1996. 1996. and mechanistic analysis. Z. Mutlib. Chem. Res. J. Troke. T. Presle. Res. 1999. Nicholson. Vanderhoeven. Kevin Park. Milzani. 1997. Hamelin. D. S. Z. Metabolic fate of pitavastatin. W. P. Toxicol. G. Chem. P. Pryde. Med. J. M.S. C. Scatina. Dean. B. li. J. Lindon. Davis. H. G. Biophys. 19. Sci. Sallustio. 2000. Jenkins. Dispos. 360 – 367. M. Khalifah. Res. Dispos.Y. Biol. A. W. Peakman. M. Prueksaritanont. J. DeGraaf. Cyclization of the acyl glucuronide metabolite of a neutral endopeptidase inhibitor to an electrophilic glutarimide: synthesis. Baynes. Meng. 50. Benet. G. V. Reactivity of mefenamic acid-1-O-acyl glucuronide with proteins in vitro and ex vivo. Wilson. P. H. G. Glucuronidation of statins in animals and humans: a novel mechanism of statin lactonization. M. NMR and QSAR studies on the transacylation reactivity of model 1b-O-acyl glucuronides. M. A. 842 – 849. 27. J. Netter. A. W. J. Toxicol. D. Drug Metab. R. I. Lapicque. J. C. Lin. Proc. F. 1206 – 1216. Intervention strategies to inhibit protein carbonylation by lipoxidation-derived reactive carbonyls. 683 – 691. Kinetic studies on the intramolecular acyl migration of b-1-O-acyl glucuronides: application to the glucuronides of (R). Glinsky. Planken. Dickinson. Hudson. C. Aldini. 73 – 85. C. 24. A. R. 3797 – 3801. V. B. Wolff. P. S. J. Qiu. B. K. E. A. 1999. Akira. Biochem. C. S. R. 5 (2008) 2317 [122] [123] [124] [125] [126] [127] [128] [129] 88. Stereoselective internal acyl migration of 1b-O-acyl glucuronides of enantiomeric 2-phenylpropionic acids. M. 53. Lindon. B. C. b) B. D. E. R. Res. 1987. Chem. Bailey. Toxicol. C. Fujino. Xenobiotica 2003. 251 – 258. J. Weekley. S. F.and (S)-hydroxyketoprofen metabolites. Shinohara. Carini. 19. Lou. Feather. Turgeon. 505 – 512. M. E. D. F. Kojima. K. Wang. Sallustio. 817 – 868. Stereoselective irreversible binding of ketoprofen glucuronides to albumin. R. 1998. 23. Dispos. X. Talaat. 141 – 146. Drug Metab. G. V. G. 1996. C. Atherton. Fang. Acad. S. Kohl. synthesis and degradation rate measurement. F. B. Pannall. I. N. Biochem. Bull. Baillie. L. J. Chem. A novel approach for predicting acyl glucuronide reactivity via Schiff base formation: development of rapidly formed peptide adducts for LC/MS/MS measurements. L. J. . Nicholson. A. Tosh. E. 27 – 41. Toxicol. Chem. 2007. C. J. Stachulski. Res. Y. Imachi.A.

[134] P. S. Hekster. Taylor. 838 – 847. 375 – 380. S. 45. M. Hansen. Yokoi. Esser. b) R. 12. Uchijima. Mortensen. Dispos. J. Bock.2318 CHEMISTRY & BIODIVERSITY – Vol. Amide N-glucuronidation of MaxiPost catalyzed by UDP-glucuronosyltransferase 2B7 in humans. in humans. J. 215 – 229. O. Elsevier. Anal. J. Staines. W. Oxford. B. Shimoda. Dispos. Lindon. [133] K. Drug Metab. R. 51 – 59. Beck. R. R. J. 1993. Huskey. M. Norikura. S.  HPLC/1H NMR spectroscopic studies of the reactive a-1-O-acyl isomer formed during acyl migration of S-naproxen b-1-O-acyl glucuronide. Chirality 1995. Y. W. Miners. 2004. 311. 1683 – 1691. 46. K. Zhang. Res. Dispos. Wen. E. 2nd edn. M. Res. 21. D. Biomed. Martin. A. 199 – 202. Nicholson. 26. Gotou. T. 2003. Drug Metab. J. K. 5 (2008) [130] C. T. Hashimoto. Bible Jr. Troke. W. Testa. J. Chem. W. A. Biochem. 1131 – 1137. B. K. C. 583 – 589. Wilson. 157 – 176. H. 34. Pharmacol. Structural characterization of anti-HIV drug candidate PA-457 [3-O-(3’. Studies on the reactivity of acyl glucuronides.3’-dimethylsuccinyl)-betulinic acid] and its acyl glucuronides in rat bile and evaluation of in vitro stability in human and animal liver microsomes and plasma. 34. C. C. T. Formation and reactivity of acyl glucuronides: the influence of chirality. 2002. 7. 30. A. R. Drug Metab. Karim. Drug Metab. A. C. D. S. Exp. Zhao. 2006. W. Smith. J. [138] P. H. Xenobiotica 1996. Y. 1013 – 1021. N1-glucuronidation and N4-acetylation of sulfadimethoxine in man. 2007. [137] M. Bigler. Klunk. A. Yang. Knights. QSAR for substituted benzoic acids in the rabbit: prediction of urinary excretion of glycine and glucuronide conjugates. Dickinson. A. 2004. Green. J. Dispos. Dispos. D. Pharmacol. L. J. H. 2002. Pharm. Species differences in N-glucuronidation. 1990. Katoh. Mortensen. Corcoran. [144] D. Martea. Eds. 14. Biochem. 2007. Dispos. D. W. B. 34. 1998. T. E. P. 26. J. H. K. S. J. S-Naproxen-b-1-O-acyl glucuronide degradation kinetic studies by stopped-flow HPLC-1H NMR and HPLC-UV. 765 – 772. S. J. Dispos. Gaganis. 35. Ohno. Potter. Lee. Watt. Nicholson. Dispos. Pharmacokinetics. Cupid. T. Tephly. 2006. Y. J. Bosch. J. Chem. Stern. W. M. Pharmacol. Pharm. [142] S. 1436 – 1442. Triggle. V. Roongta. A. El Mouelhi. Green. 545 – 551. 1993. B. 2007. Weekbld. Immunotoxicology. Beneken Kolmer. J.-H. Stereoselective glucuronidation and hydroxylation of etodolac by UGT1A9 and CYP2C9 in man. T. Xenobiotica 2004. 1167– 1176. J. E. in ADME-Tox Approaches. Rapid internal acyl migration and protein binding of synthetic probenecid glucuronides. J. R. R. Drug Metab. tranilast: identification of human UDP-glucuronosyltransferase isoforms and effect of its phase I metabolite. 403 – 415. W. U. Yoshikawa. Glucuronidation of amine substrates by purified and expressed UDPglucuronosyltransferase proteins. [131] T. C. A. Sidelmann.. King. Burchell. J. 860 – 867. Ed. 1043 – 1047. J. K. van de Waterbeemd. [147] M. D. Lampe. Drug Metab. Pharmacol. valdecoxib. Beddell. Tougou. Disposition of a specific cyclooxygenase-2 inhibitor. Sci. 73. [143] A. N-Glucuronidation of carbamazepine in human tissues is mediated by UGT2B7. H. 15. Nicholson. 449 – 461. D. 2001. [132] B. [135] J. [146] T. Matsui. Dickinson. D. H. Kuehl. G. 1 – 9. G. G. King. Cornett. Eds. I. M. Glucuronidation of fenamates: kinetic studies using human kidney cortical microsomes and recombinant UDP-glucuronosyltransferases (UGT) 1A9 and 2B7. T. [145] J. R. [141] Z. Akira. 5 in Comprehensive Medicinal Chemistry. Vol. Lee Chiu. Lindon. p. G. King. Coughtrie. O. Nakamura. Drug Metab. Ther. Biochem. Mitroka. [139] K. 1363 – 1370. c) O. Covalent binding of diflunisal to tissues of the rat. 31. Zhang. Corcoran. Stereoselective glucuronidation of (R). Hayball. 26. Zhu. Contrasting systemic stabilities of the acyl and phenolic glucuronides of diflunisal in the rat. 29. 2001. R. Vree.-C. . 32. Glucuronidation of antiallergic drug. C. J. G. [136] G. M. K. IV. Yuan. D. W. Drug Metab.. Xenobiotica 1991. R. Findlay. Rapid LC-TOFMS method for identification of binding sites of covalent acylglucuronides-albumin complexes. Toxicol. R. 1298 – 1300. Ohkawa. Toxicol. 1998. J.and (S)-naproxen by recombinant rat phenol UDP-glucuronosyltransferase (UGT1A1) and its human orthologue. Hansen. Glucuronidation of the aspirin metabolite salicylic acid by expressed UDP-glucuronosyltransferases and human liver microsomes. [140] a) M.

D. B. V. E. Lakshmi. Luo. F. K. K. Lakshmi. Babu. 431 – 441. R. T. 27. G. Davis. Oesch. Pharmacol. 30. S. Babu. E. by human liver microsomes and UDP-glucuronosyltransferase 1A4. R. 2004. 5 in Comprehensive Medicinal Chemistry.. Herrera. Zenser. 507 – 512. D. J. B. in ADME-Tox Approaches. Elsevier. Midha. B. 1281 – 1288. Oxford. V. 26. and their hydroxy metabolites. Rev. Ohnuma. McNelly. M. 19. B. B.. Hiratsuka. A. H. B. U. Muro. R. a general phenomenon in the metabolism of H1-antihistamines in humans. Tertiary N-glucuronides of clozapine and its metabolite desmethylclozapine in patient urine. Drug Metab. McKay. N. J. Carcinogenesis 1994. Dispos. Testa. V. M. Kapetanovic. Ethnic differences in N-glucuronidation of nicotine and cotinine. 1998. Hiratsuka. Pharmacol. Eds. Ohnuma. Ghosheh. Nelson. 1999. Dispos. E. Glucuronide conjugates of 4-aminobiphenyl and its N-hydroxy metabolites. O. N-Glucuronidation of nicotine and cotinine by human liver microsomes and heterologously expressed UDP-glucuronosyltransferases. Adv. S. 26. H. G. Luo. The Metabolism of Drugs and Other Xenobiotics – Biochemistry of Redox Reactions. P. 26. K. 87 – 132. Drug Metab. Davis. Huang. B. Lazarus. H. D. Drug Delivery Rev. 21. Microsomal N-glucuronidation of nicotine and cotinine: human hepatic interindividual. T. Owens. M. 1999. in ADME-Tox Approaches. M. 5 in Comprehensive Medicinal Chemistry. Mojarrabi. Davis. Drug Metab. 2nd edn. 2093 – 2102. T. 1478 – 1483. V. Nishiyama. Hawes. Eds. Fong. I. C. J. Kuehl. Hawes. Pharmacol. 31. 26. 1998. K. T. Lakshmi. F. 1991. P. 51. Y. E. Hawes. I. V. 1990. The metabolic fate of amitriptyline. 593 – 599. Drug Metab. Dispos. 722 – 724. I. A. N þ -Glucuronidation. M. Ogura. Characterization of Nglucuronidation of the lung carcinogen 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanol (NNAL) . Midha. U. Xenobiotica 1991. K. E. 29. M. E. Biochem. Oxford. T. E. Vol. S. Anderson. 1996. P. E. Ther. 5 (2008) 2319 [148] [149] [150] [151] [152] [153] [154] [155] [156] [157] [158] [159] B. E. G. McKay. T. 36. pH stability and synthesis by human and dog liver. 193 – 214. B. L. p. 856 – 859. E. 723 – 746. H. Taylor. B. B. Davis. 1679 – 1685. J. 2003 – 2007. Doerge. Elsevier. Kaku. Dispos. V. a new anticoagulant. D. unusual metabolites of gallopamil and verapamil. Trager. D. Exp. Dispos. Hawes. Pharmacol. 2003. Tephly. Perez-Stable. The quaternary ammonium-linked glucuronide of doxepin: a major metabolite in depressed patients treated with doxepin. Fang. 67. I. Zenser. a common pathway in the human metabolism of drugs with a tertiary amine group. M. 1343 – 1348. 1998. Role of pharmacokinetics in the discovery and development of indinavir. 1064 – 1067. human intertissue. V. K. B. 2006. Triggle. 33 – 49. T. B. Eds. M. Murphy. Triggle. p. Zenser. Oesch-Bartlomowicz. Quaternary ammoniumlinked glucuronidation of tamoxifen by human liver microsomes and UDP-glucuronosyltransferase 1A4.  Human and Escherichia coli b-glucuronidase hydrolysis of glucuronide conjugates of benzidine and 4-aminobiphenyl. 1998. Pharmacol. J. T. Biochem. Ogura. nortriptyline and amitriptylineoxide in man. Breyer-Pfaff. by human liver microsomes and liver slices. D. P. J. W. Korchinski. G. T. 2nd edn. Biochem. Lin. In vitro glucuronidation of D-23129. Zenser. Wiener. P. Taylor. Academic Press. Ther. 1196 – 1203. T. K. K. Kaku. Drug Metab. J. J. Mutlib. Testa. Quaternary ammonium-linked glucuronidation of trans-4hydroxytamoxifen. 830 – 837. B. J. W. Benowitz. Dispos. M. Glucuronidation of amines and other xenobiotics catalyzed by expressed human UDP-glucuronosyltransferase 1A3. Korchinski.-L. L. Vol. Breyer-Pfaff. 1361 – 1368. 2004. Dispos. 2007. 2007. and interspecies hepatic variation. London. Upadhyaya. J. Strong. Testa. Muro. Exp. Drug Metab. Kupferberg. V. A. Synthesis and identification of the N-glucuronides of norgallopamil and norverapamil. Mackensie. M. Lakshmi.  Human liver glucuronidation of benzidine. van de Waterbeemd. Torchin. W. B. J. D. 39. B. S. 15. 291. Dispos. J. Xenobiotica 1997. Jacob III. Drug Metab. H. 2002. Drug Metab. 71. N-Glucuronidation of benzidine and its metabolites. Mechanisms of toxification and detoxification that challenge drug candidates and drugs. R. Principles of drug metabolism 1: Redox reactions. Ishikawa. N þ -Glucuronidation of aliphatic tertiary amines. G. Nishiyama. L. Eds. 1358 – 1369. T. 1995. 2001. 252.CHEMISTRY & BIODIVERSITY – Vol. H. P. van de Waterbeemd. modin. Drug Metab.

H. J. Pharmacol. A. Elmarakby. Gauthier. 2004. Nicholson. R. 2006. Quaternary ammonium-linked glucuronidation of 1-substituted imidazoles: studies of human UDP-glucuronosyltransferases involved and substrate specificities. methylbiphenyl triazole. W. Dispos. Voihsel. Miller. 1010 – 1016. Relative Nglucuronidation reactivity of methylbiphenyl tetrazole. D. L. Drug Metab. 34. Masucci. S. J. R. T. J. P. . Drug Metab. Carmella. J. Upadhyaya. 651 – 658. M. 29. tioconazole. N-glucuronide metabolite of the imidazole antifungal. Macrae.and O-glucuronides of 4(methylnitrosoamino)-1-(3-pyridyl)-1-butanol (NNAL) in human urine. D. E. Drug Metab. Dispos. 1993. Zhang. O. G. A. Glucuronidation of the lung carcinogen 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanol (NNAL) in rat UDP-glucuronosyltransferase 2B1. H. D. 126. S. Miners. S. Yan. M. Acta 1996. 2001. A. Q. V.. Tarbit. Dispos. 1994. M. C. 545 – 550. O. G. Mallis. L. 72 – 79. F. N-Glucuronidation reactions.-H.4-dihydroindeno[1. Hawes. Mackenzie. Dispos. Jones. Braun. 2002. Brenner-Weiss. Mackenzie. Drug Metab. II. Dispos. N-Glucuronidation of the platelet-derived growth factor receptor tyrosine kinase inhibitor 6. I. F. Doss. 1990. E. 855 – 869. novel. 1950 – 1954. McConnell. Science 1980. S. Giddings. R. 1050 – 1056. P. Pharmacol. Toxicol. Dispos. Dieterle. Oellerich. 2002. Y. A. Lu. 22. I. R.2320 CHEMISTRY & BIODIVERSITY – Vol. 792 – 799. 20. NMR characterization of an S-linked glucuronide metabolite of the potent. W. Detection of mono. D. 2006. F. Sulfinpyrazone C-glucuronidation is catalyzed selectively by human UGT-glucuronosyltransferase 1A9. Dispos. J. G. Kamimura. L. C. 21. Braun. V. In vitro characterization of lamotrigine N2-glucuronidation and the lamotrigine-valproic acid interaction. R. Takahara. 1075 – 1082. Mei. Biochim. L. W. Elliot. 1290 – 1295. Nakaoka. Xenobiotica 2003. 703 – 704. I. W. P. J. Uetrecht.2-c]pyrazol-3-yl)-(3fluorophenyl)-amine by human UDP-glucuronosyltransferases. Drug Metab. Eur. nonsteroidal progesterone agonist tanaproget. R. 30. Hawes. D. C. monkeys and humans. Dispos.-H. 1999. R. C. P. C-Glucuronidation of the acetylenic moiety of ethchlorvynol in the rabbit. Le. Possible bioactivation pathways of lamotrigine. H. Keating. 33. W. Wang. Drug Metab. Lazarus. 9. Y. Murphy. Xenobiotica 1990. Identification of glucoside and carboxyl-linked glucuronide conjugates of mycophenolic acid in plasma of transplant recipients treated with mycophenolate mofetil. M. Chandrasekaran. Ghosheh. D. Kinetic characteristics of UGT-glucuronosyltransferases towards a dithiol metabolite of malotilate in hepatic microsomes of rats and rabbits. T. Z. R. Schoen. G. Faigle. Gruenke. D. Johnson. 748 – 755. S. P. J. W. 34. Lee Chiu. Clin. J. Miners. Drug Metab. H. 619 – 627. Feil. Castro-Perez. N. McKay. dogs. Lamoureux. S. Rowland. Paulson. Drug Metab. W. G. J. L. H. W. Shen. Lactose conjugation of sulphonamide drugs in the lactating dairy cow. 209. Hecht. 27. S. Park. F. Dannenberg. W. 20. Huskey. Caldwell. 1283 – 1287. J. J. 1100 – 1102. Niedmann. 1999. Strassburg. S. Elliot. Huskey. Ho. J. Craig. Shipkova. Kerdpin. 309 – 315. M. Res. J.and di-hexoses as metabolites of 4-bromoaniline using HPLC-TOF-MS/MS. Xenobiotica 1992. Wieland. Olson. M. Vashishtha. 2006.7-(dimethoxy-2. C. P. M. A. Pullen. 5 (2008) [160] [161] [162] [163] [164] [165] [166] [167] [168] [169] [170] [171] [172] in human liver: importance of UDP-glucuronosyltransferase 1A4. Shipkova. A. Dickinson. R. J. and methylphenyl imidazole in rat. Chem. Tozer. O. Miller. W. D. P. 1289. J. Dispos. Quaternary ammonium-linked glucuronidation of 1-substituted imidazoles by liver microsomes: interspecies differences and structure-metabolism relationships.-H. J. E. O. DeMaio. 18. Theobald. Analysis of N. Tuman. 925 – 939. W. 1975.-H. Barua. McCann. 15. Williams. J. Leon. D. Ogata. Bidlack. K. W. Schütz. Abolin. Streit. C. Mory. G. Drug Metab. Lee Chiu. M. E. Tephly. C. S. A. Hogg. G. J. and human microsomes. A. R. 2007. Wilson. J. 34. J. E. 35. O. 1070 – 1076. Richter. Biophys. Vashishtha. M. D. W. monkey. Drug Metab. Formation of b-glucuronides and of bgalacturonides of various retinoids catalyzed by induced and noninduced microsomal UDPglucuronosyltransferases of rat liver. Genchi. P. M. 22. C. Kinns. K. S. 2006. I. Dispos. McCann. Major. S. N-Glucuronidation reactions. Drug Metab. Dispos. W. J. H. H. Ren. Br. A. K. 34. Galemmo Jr. Tetrazole Nglucuronidation of selected angiotensin II receptor antagonists in hepatic microsomes from rats. V. 284 – 290. W. Biotransformation and pharmacokinetics of sulfinpyrazone (Anturan) in man. Characterization of a quaternary. R. Dispos. A. Drug Metab. G. 1055 – 1062. 135 – 145. 32. J. Y. Drug Metab. S. 1992. J. Armstrong. a-Glucoside formation of xenobiotics by rat liver aglucosidases. G. Dispos.

S. M. M.4]triazol-3-yl)pyridine-2-carbonitrile) in the rat. J. Principles of Organic Stereochemistry. Uridine diphosphate sugar-selective conjugation of an aldose reductase inhibitor (AS-3201) by UDP-glucuronosyltransferase 2B subfamily in human liver microsomes. T. monkey. Toide. Harvey. D. Biochem. 2005. K. Pharmacol. glutathione S-transferase. Identification of glucose conjugates as major urinary metabolites of cannabidiol in the dog. D. Tukey. Oellerich. J. T. Res. Abraham. M. N. Dispos. S. 291 – 292 (http://louisville. Rev. S. F. H. 1990. 37. C.html). Mongrain. J. J. Sjçberg. 2001. 42. 58. D. 2004. 470 – 472. Ueo. Y. Roots. 1027 – 1034. S. Tsuji. G. 1995. Metabolic profile of FYX-051 (4-(5pyridin-4-yl-1H-[1. Trent. Noguchi. Pharmacol. New York. 59 – 124. A. Rev. M. V. Vermeulen. Boukouvala. Samara. A. 6. 1880 – 1886. Uwai. Med. J. and humans: identification of N-glucuronides and N-glucosides. 157 – 234. 2007. Grewal. Gaedigk. 2007. J. J. Pharm. Pharmacogenetics 2000. Fujii. M. Liu.  Human acetyltransferase polymorphism. 5 (2008) 2321 [173] [174] [175] [176] [177] [178] [179] [180] [181] [182] [183] [184] H. 1996. Soine. N-Acetyltransferase. J. Br. W. M. E. 1997. D. E. 1300 – 1308. K. 132. Eukaryotic arylamine N-acetyltransferase. Drug Metab. Identification of the diastereomers of pentobarbital N-glucosides excreted in human urine. N. 1468 – 1483. Bialer. Graham. Stereochemical characterization of the diastereomers of the amobarbital N-glucosides excreted in human urine. J. Drug Metab. Soine. I. R. Arylamine N-acetyltransferases: characterization of the substrate specificities and molecular interactions of environmental arylamines with human NAT1 and NAT2. H. Hansen. Ther. Katsura. E. Price-Evans. Pharmacol. Y. P. Dupret. A. Res. D. Pharmacol. Duggan. Drug Metab. 347 – 359. D. L. Cornett. Res. T. a) L. W. Commandeur. Glucosidation as a new conjugation pathway for metabolites of bis(2ethylhexyl) phthalate. 1992. dog. Soine. P. 97. Hein. R. R. 1999. J. Biochem. Dispos. Yamazaki. Graham. H. Itogawa. 29. M. Pharm. Chem. A. Interaction and transport characteristics of mycophenolic acid and its glucuronide via human organic anion transporters hOAT1 and hOAT3. Walraven. Armstrong. D. G. H. K. W. Brockmçller. E. Walters. Westkaemper. H. England. 1001 – 1007. H. 34. Thomas. H. P. H. 2008. I. Motohashi. 67. W. Soine. Hughes. P. B. 2002. 1990. Drug Metab. Computational and experimental analyses of mammalian arylamine N-acetyltransferase structure and function. Hein. A. 161 – 168. 11. J. 19. S. Nagata. 1535 – 1539. D. 376. Rhodes. 1989. J. 1759 – 1764. P. Wagner. Dispos. W. New metabolites of the drug 5-aminosalicylic acid: N-b-dglucopyranosyl-5-aminosalicylic acid. 402 – 406. D. England. Wormhoudt. S. 190. T. 511 – 564. 23. J. R. Grçne. T. Pharm. C. Pharmacol. D. Cao. Soine. 74. Grant. Terauchi. J. Update on consensus arylamine N-acetyltransferase gene nomenclature. A. 563 – 569. Stereochemical characterization of the diastereomers of the phenobarbital N-b-d-glucose conjugate excreted in human urine. Bell. Exp. W. M. Glucuronide and glucoside conjugation of mycophenolic acid in plasma by human liver. b) D. Tjørnelund. 61 – 70. Kawamura. Weber. N.-H. 891 – 899. Egestad. 548 – 552. E. kidney and intestinal microsomes. Genetic polymorphisms of human Nacetyltransferase. Janezic. 2006. M. Wagner. J. L. T. Vatsis. Toxicol. W. R. K. D. Identification of phenobarbital N-glucuronides as urinary metabolites of phenobarbital in mice. W. Price-Evans. Tsiftsoglou. Grant. Iwanaga. 35. D. Pharmacol. 794 – 800. Yu. A. Investigation of substrate specificity by highthroughput screening. J. G. B. Wieland. E.-J. Cascorbi. T.2. Crit. Biochem. Yu. M. and epoxide hydrolase enzymes: relevance to xenobiotic metabolism and toxicity. cytochrome P450. O. 69. J. H. Res. A. 20. Soine. Kurono.-F.CHEMISTRY & BIODIVERSITY – Vol. 20. N. Stability and hydrolysis kinetics of spirosuccinimide type inhibitors of aldose reductase in aqueous solution and retardation of their hydrolysis by the target enzyme. S. Mutat. P. Res. Pharm. Nakazawa. Goodfellow. Baldwin. C. Hanna. Capps. Chem. Vath. Xenobiotica 1990. W. R. 1999. N-Glucoside formation as a detoxification mechanism in mammals. NGlucosylation of phenobarbital analogs by mouse liver microsomes. Borchert. O. C. J. 2007. Arylamine N-acetyltransferases: what we learn from genes and genomes. 174 – 189. . T. Sim. 177 – 183. Fakis. Wireko. Pharmacol. M. E. L. C. 7. Drug Metab. K. Soine. Dispos. P. Toxicol. H. H. c) C. V. P. Sci. Kamataki. 10. Y. G. 1994. Soine. Hanna. W. B. H. W. 1979. Williams. 1269 – 1278. Neighbors. H. E. Von Vett. M. Omura. K. H. H. Inui. E. Correlation between genotype and phenotype of the human arylamine N-acetyltransferase type I (NAT1). Testa.-M. Chen. A. Dekker. Sim. Ther. Arison. Xenobiotica 1989. Mushtaq. C. A. B. Res. R. Zjang. Bruhn. 1974. M. 7. J. 20. Sanjoba. Biochem.

Isoniazid: metabolic aspects and toxicological correlates. Nomenclature for N-acetyltransferases. Suzuki. 13 (Suppl. S. 63. 8. Nishio. P. The influence of dose and Nacetyltransferase-2 (NAT2) genotype and phenotype on the pharmacokinetics and pharmacodynamics of isoniazid. A. B. H. Chen. Jitsufuchi. J. Pharmacol. D. Polymorphism of the N-acetyltransferase 2 gene as a susceptibility risk factor for antituberculosis drug-induced hepatitis. S. Dispos. P. Meyer. Hepatology 2002. H. J. L. 156 – 170. Acetylation and its role in the mutagenicity of the antihypertensive agent hydralazine. Ed. Lemke. Hwang. Abramocivi. Grant. M. N-Acetylcyanamide. a metabolite of levosimendan. 1990. Biol. W. K. W. M. 560 – 564. H. Nagasawa. I. W. Pharmacol. Lin. Y. Nat. Preziosi. Parkin. C. A.2322 CHEMISTRY & BIODIVERSITY – Vol. Dispos. 23. M. Y. L. Structure of arylamine Nacetyltransferase reveals a catalytic triad. AcNDP. R. E. S. Ieiri. P. 35. J. 345 – 351. Lee. A. Drug Metab. D. G. Biol. 5. E. Noble. Clin. 5 (2008) [185] [186] [187] [188] [189] [190] [191] [192] [193] [194] [195] [196] Grant. L. R. B. 1697 – 1702. Carreon. Ono. Maritz. A. Werely. Kudo. T. Wu. Nikkanen. Dispos. McQueen. 30189 – 30197. J. M. Cole. D. 48. V. Sirgel. Martin. A. Grant. 254 – 260. Y. 213 – 222. S. Zonisamide: chemistry. 1995. Metabolism of OR-1896. Pharmacol. Mitchell. Determination and properties of acetyl conjugate of N-desisopropylpropranolol. 1270 – 1275. Drug Metab. mechanism of action. Estrada-Rodgers. Donald. M. Sim. D. Drug Metab. E. H. Wu. 34. S. Curr. W. Yu. Levy. U. Huang. Noda. . Dispos. F. 2007. Clin. H. 1995. 1064 – 1070. D. van Heiden. H. Noda. N. Wu. Johnston. T. J. Substrate selectivity of mouse N-acetyltransferases 1. acetylhydrazine. Br. Morioka. Drug Metab. H. Eto. Dispos. 888 – 894. Structural basis of substrate-binding specificity of human arylamine N-acetyltransferase. Chem. Drug Metab. acetylisoniazid. D. Pharmacokinetic study of zonisamide in patients undergoing brain surgery. M. and man. F. T. 2007. Drug Metab. H. Hekster. Determination of isoniazid and its hydrazino metabolites. D. D. N. Schulte. M. 1997. N. N. Y. Dombrovsky. H. 337 – 344. 2004. M. 1984. 1). S. N. Rozmahel. rabbit. C. Brenneman. R. H. T. 431 – 438. S. L. J. Further evidence for an acetylator phenotype difference in the metabolism of hydralazine in man. 852 – 858. Antila. Angew. P. 502 – 505. Pesonen. 34. 927 – 933. Yang. Drug Metab. Sugamori. R. 18. Kubota. Vuorela. P. E. Hayes. Kim. Pharm. Y. N. J. 23. J. S. Stiff. N. 18. C. M. and diacetylhydrazine in human plasma by GC-MC. V. C. J. Sandy. Ruder. 1454 – 1455. the major urinary metabolite of cyanamide in rat. Grant. Struct. S. T. Beneken Kolmer. J. Wong. R. Venter. Shimoda. K. and N4-acetylation of sulfa-6-monomethoxine in humans. D. 2006. Dalton. Kwon. Gao. and 3 expressed in COS-1 cells. Hein. Smith. in rats and humans. 2007. 2007. Seizure 2004. S. Hasunuma. D. V. J. M. 839 – 851. 2007. Eur. Leppik. K. J. R. Higuchi. Facchini. M. R. L. Miura. Pharm. and pharmacokinetics. Shirota. Pharmacol. Sim. S. Dispos. D. Ono. J. Dispos. J. N1-glucuronidation. M. Koskinen. Scheinin. Iijima. 1996. Seifart. Grant. Vree. H. Zemaitis. Miller. Brennerman. Lauterburg. 559 – 565. Bull. B. Wang. 883 – 889. Boissy. 2. A. V. P. E. D. Eur. Clin. H. Delgoda. 2000. Effect of arylamine acetyltransferase Nat3 gene knockout on N-acetylation in the mouse. 1990. R. A. Metabolism of the anticonvulsant agent zonisamide in the rat. S. 26. T. J. Chromatogr. Pharmacogenetics 1995. 38. Fukui. Lotta. S. Goodfellow. P. W. Pharmacokinetics. 282. Pykäläinen. Waters. S. Sinclair. Rothman. M. 633 – 639. Chem. Su. A. Drug Metab. C. Y. Gaedigk. W. A. J. 11. 2006. Lehtonen. Thompson. Demaster. G. A. In vivo and in vitro metabolism of arylamine procarcinogens in acetyltransferase-deficient mice. S5 – S10. Chang. Biol. Drug Metab. Schaaf. 46. D. H. Dose-escalation study of isoniazid in healthy volunteers with the rapid acetylator genotype of arylamine N-acetyltransferase 2. L. Lai. J. W. J. Dispos. A. Yamazoe. Azuma. I. Xenobiotica 2008. C. F. 2007. E. D. 7. 7621 – 7624. A selective chemical probe for coenzyme A-requiring enzymes. Int. I. Weber. Sci. Loppnau. J. Jiang. P. 1981. Plotnikov. Puttonen. Relling. 63. 12. E. Interspecies differences in pharmacokinetics and metabolism of S-3-(4-acetylamino-phenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3-trifluoromethylphenyl)-propionamide: the role of N-acetyltransferase. T. 1981. D. Tapanainen. A. J. K. 18. T. Eur. Timbrell. H. L. 224. M. A. J. Sugamori. L. G.. A. F. U. J. M. Tempel. 1 – 17. Pharm. dog. P. Pharmacokinetics of levosimendan and its active metabolite OR-1896 in rapid and slow acetylators. Vaahtera. Y. X. 35. S. Ohno. Kelleher.

711 – 720. L. Chem. Buhman. Metabolic activation of N-hydroxy-2aminofluorene and N-hydroxy-2-acetylaminofluorene by monomorphic N-acetyltransferase (NAT1) and polymorphic N-acetyltransferase (NAT2) in colon cytosols of Syrian hamsters congenic at the NAT2 locus. Drogemuller. Bioactivation of Nhydroxy-2-acetylaminofluorenes by N. Zenser. R. G. Goldman. F. Biochem. Dahlbäck. 44. Josephy. 1986. R. F. 1667 – 1672. 1311 – 1317. 1689 – 1697. 203 – 214. Varwell Marsh. P. Toxicol. R. G. Jackowski. Mammalian acyl-CoA : cholesterol acyltransferases. Ultimate carcinogenic metabolites from aromatic and heterocyclic aromatic amine: a computational study in relation to their mutagenic potency. R. Zhang. P. Res. K. F. Cancer Res. Boteju. an antiasthma glucocorticoid. Wagner. 1983. Reversible formation of fatty acid esters of budesonide. Chem. Biochim.Oacetyltransferase and arylamine N-acetyltransferase. A. Conney. 1995. Boteju. Pharmacokinetics of budesonide and its major ester metabolite after inhalation and intravenous administration of budesonide in the rat. Khan. K. 26. Res. J. Kaphalia. V. F. Toxicol. R. Rock. A. Rev. W. C. S. Drug Metab. 49 – 66. G. 2006. Stimulatory effect of clofibrate and gemfibrozil administration on the formation of fatty acid esters of estradiol by rat liver microsomes. H. F. Saito. 861. Toxicol. 684 – 689. Dodds. Lipid Res. Interrelationships between xenobiotic metabolism and lipid biosynthesis. M. Hochberg. Tunek. 14. Prog. Arylamine-nucleoside adduct formation: evidence for arylnitrene involvement in the reactions of an N-acetoxyarylamine.-J. 2007. Lau. D. 188 – 197. 2000. Accad. NHydroxyarylamine O-acetyltransferase in hamster liver: identity with arylhydroxamic acid N. A. Fatty acid conjugates of xenobiotics. A. Davis. LeMasters. A. Hein. Reversible fatty acid conjugation of budesonide. A. 1994. Inter-relationships between xenobiotic metabolism and lipid biochemistry. G. 161 – 168.. 180 – 186. Cancer 2006. E. 29. Hanna. 331 – 348. Interact. Kato. 20. Res. Hallstrçm. Brass. A. Pharmacol. K. Acta 2000. 1529. Eds. 70 – 74. 118. B. M. Metabolic activation of aromatic amine mutagens by simultaneous expression of human cytochrome P450 1A2. 2005. C. F. H. Prog. L. Hanna. Lett. P. 769 – 776. Chem. Jendbro. E. Falk-Nilsson. Rustan. B. 1985. Parikh. 5 (2008) 2323 [197] [198] [199] [200] [201] [202] [203] [204] [205] [206] D. E. K. 34.O-acyltransferase: substituent effects on covalent binding to DNA. NAT2 slow acetylation and bladder cancer in workers exposed to benzidine. E. 1995. S. V. Toxicol. 1995. Strandberg. Yin. V. Kamataki. 623 – 630. Lightstone. 7. J. Borosky. Res. P. Zhu. Drug Metab. A. H. S. B. J. Wang. M. H. G. J. Caldwell. 1994. B. 509 – 514. 2001. J. 1651 – 1657. C. J. S. 1998. Dispos. P. M. A. C. 32. Spencer. M. 25. 183 – 197. R. K. R. Farese. Felton. and N-acetyltransferase in E. O. Biological esterification of steroids. Insights into the O-acetylation reaction of hydroxylated heterocyclic amines by human N-acetyltransferases: a computational study. S. J. K. 296. Chem. E. Drug Metab. L. 136. B. 99. T. G. 1. 142 – 154. L. Chem. C. Dispos. Miller-Larsson. Kadlubar. 1997. D. 53. Ansari. 2001. M. R. K. 90. Biochem. Ther. W. F. Carcinogenesis 1993. Role of hepatic fatty acid : coenzyme A . Endocrinology 1995. 1 – 17. P. D. Sjçdin. Endocr. Lipid Res. Shinohara. Int. 219 – 247. Chem. Lakshmi. 1182 – 1190. 1993. 2005. Gupta. Coenzyme A: back in action. Evans. W. P. G. Brattsand. Knights. Drug Metab. NADPH-cytochrome P450 reductase. Dispos. Xu. T. D. 19. Mattsson. T. Hjertberg. S.-Biol. Overview of coenzyme A metabolism and its role in cellular toxicity. Edsbäcker. P. F. 125 – 153. Knights. C. Bell. 19. Toxicol. Tunek. Y. Hsu. Biophys. in human lung and liver microsomes. Pharmacol. coli. M. Doll. 847 – 862. 1998. 18. T. Caldwell. 8. P. The role of acetylation in benzidine metabolism and DNA adduct formation in dog and rat liver. Res. G. Gray. E. 75. Biochem. B. Curr. Soc. 171 – 180. Ferguson. H. M. Hochberg. Res. M. A comparison of the esterification of steroids by rat lecithin : cholesterol acyltransferase and acyl coenzyme A : cholesterol acyltransferase. 1998. S. J. Trans. Hanna. Toxicol. S. Chem. Exp. Pahija.CHEMISTRY & BIODIVERSITY – Vol. H. Hemstreet III. D. Leonardi. Y.-M. 11. Toxicol. Xenobiotic-CoA ligases: Kinetic and molecular characterization. Xenobiotic lipids: the inclusion of xenobiotic compounds in pathways of lipid biosynthesis. Guengerich. F. Lake. R. Johansson. Irreversible inactivation of arylkamine N-acetyltransferases in the presence of N-hydroxy-4-acetylaminobiphenyl: a comparison of human and hamster enzymes. B.

1992. E. M. 44. Clin. J. Z. N. Roy-de Vos. J. M. J. Coenzyme A esters of 2-aryloxyphenoxypropionate herbicides and 2arylpropionate antiinflammatory drugs are potent and stereoselective inhibitors of rat liver acetylCoA carboxylase.-Y. Biophys. Stereoselective and nonstereoselective effects of ibuprofen enantiomers on mitochondrial b-oxidation of fatty acids. Hansen. 1997. Etter. Sidenius. b) R. Pharmacol. Sallustio. J. Chem. Exp.2324 CHEMISTRY & BIODIVERSITY – Vol. 159 – 168. C. Alexson. F. 15. Toxicol. J. 18. Benoit. Brune. 2029 – 2032. Pharmacol. 1990. Amigo. J. 2005. Letteron. 758 – 764. W. The biochemistry and toxicology of benzoic acid metabolism and its relationship to the elimination of waste nitrogen. Yamada. J. I. 1985. Biol. Degott. Igarashi. K. Biochim. M. Sidenius. C. Int. J. 533 – 540. Larrey. M. Acta 1992. A. and palmitoylCoA conjugation in vitro: kinetic and molecular characterization of marmoset liver microsomes and expressed MLCL1. Levoin. Olsen. 1995. Knights. Taylor & Francis. M. Dispos. J. S. Biochem. 1124. Benet. Caldwell. Y. G. C. Freneaux. MacLeod. Testa. Sykes. B. Z. 1986. 3. Bjørnsdottir. P. Fukui. 487 – 492. E. M. G. Hilal. C. C. G. E. D. M. 529 – 535. Olsen. E. Chem. 2002. Long-chain acyl-CoA hydrolase in the brain. Yamaoka. I. Studies on the metabolism of tolmetin to the chemically reactive acyl-coenzyme A thioester intermediate in rats. K. I. 1993. 25. Ther. Drogemuller. 1992. M. C. 2002. A. J. L. Drew. Roberts. Kasuya. C. 75 – 81. M. ciprofibroyl-. 1994. S. Jouzeau. Blondeau. L. Masters. Br. J. 145 – 150. Ed. Brugger. a) J. Skonberg. J. 163. K. Amino acid conjugation. Pharmacol. S. R. C. 35. Waku. 2007. J. Hunt. Singh. Li. Physiol. Biochem. M. 273 – 278. I. B. 2004. Res. L. Chapleur. 255. Y. Berson. Hunt. 2001. M. S. J. Sidenius. Fromenty. Pessayre. 776 – 782. Bronfman. In vitro reactivity of carboxylic acid-CoA thioesters with glutathione. Alexson. Res. 1998. Enantioselective binding of 2-phenylpropionic acid to protein in vitro in rat hepatocytes. Grandcolas. 60. Elucidation of the mechanism of inhibition of cyclooxygenases by acyl-CoA and acyl glucuronide conjugates of ketoprofen. C. K. N. Amino Acids 2005. J. Lapicque. K. C. J. Activation of hypolipidaemic drugs to acyl-CoA thioesters. 22. Hutt. 176 – 182. Chem. Hansen. Benet. Biochem. Drogemuller. Nafenopin-. Bjørnsdottir. Miners. Toxicol. Labbe. 2007. Li. K. J. C. b) J. J. C. Toxicol. 46. 667 – 673. Characterization of an acyl-CoA thioesterase that functions as a major regulator of peroxisomal lipid metabolism. 1998. 1729 – 1736. L. Knights. Chem. Ther. Chem. J. Chretien. Cl. K. J. 273 – 305. 49. Neupert. Pharmacol. C. Cassida. Biochem. Lipid Res. Kinetic characterization of rat liver peroxisomal nafenopin-CoA ligase. U. in Conjugation Reactions in Drug Metabolism. C. S. Toxicol. Pharmacol. 1480 – 1487. Effects of ibuprofen enantiomers and its coenzyme A thioesters on human prostaglandin endoperoxide synthases. 239. K. Wright. R. F. M. J. Expert Opin. 2002. H. 1291 – 1296. El-Aaser. Nunthasomboon. F. Knights. 1335 – 1339. A revised nomenclature for mammalian acyl-CoA thioesterases/hydrolases. Origins and fates of fatty acyl-CoA esters. Life Sci. Yamada. K. 2005. Appl. Knights. Molecular specificity of a medium chain acyl-CoA synthetase for substrates and inhibitors. . 2004. Li. J. 396. In vitro covalent binding of nafenopin-CoA to human liver proteins. Biochem. S. Res. M. 59 – 72. L. H. Pharmacol. B. Guillaume. Interactions of anti-inflammatory 2-arylpropionates (profens) with the metabolism of fatty acids – in vitro studies. 56 – 64. L. P. Geisslinger. Kemal. Knights. Benet. P. On the role of the peroxisome in the metabolism of drugs and xenobiotics. Grillo. Morales. Tissue React. H. London. 63 – 90. D. Amino acid conjugation: contribution to the metabolism and toxicity of xenobiotic carboxylic acids. 1957 – 1969. J. U. Biochem. Exp. B. M. B. Biophys. Netter. Pharmacol. 781 – 784. p. 55. Drug Metab. E. 101 – 111. Skonberg. Mulder. 1998. A. Kase. a) U. H. Euchenhofer. 1990. Pharmacol. 28. Olsen. Z. Mayer. 1769 – 1775. Solaas. 17. A comparative quantum-chemical study of methyl acetate and S-methyl thioacetate. M. Podesta. Maltais. J. Pharmacol. Toxicol. Nunthasomboon. 1128 – 1138. C. G. 56. Hansen. Biochem. 5 (2008) [207] [208] [209] [210] [211] [212] [213] [214] [215] [216] [217] [218] ligases in the metabolism of xenobiotic carboxylic acids. Arch. Drug Metab. O. 50. M. E. S. 68. C. C. The effects of ibuprofen enantiomers on hepatocyte intermediary metabolism and mitochondrial respiration. K. Audergon. Pharmacol. Biophys. A. Toward an understanding of the biochemical reactivity of esters of coenzyme A. F. 16. Qureshi. J. In vitro and in vivo studies on acyl-coenzyme A-dependent bioactivation of zomepirac in rats. W. 277. 122. Tremblay.

[227] a) M. J. Pigott. Drug Metab. 1996. S. Nakamura. K.-Act. R. J. Life Sci. G. J. [230] S. H. C. A. Franklin. Notarianni. 49. Shen. 805 – 809. 1166 – 1175. W. 1996. Mauleon. Mohri. 3D-Quantitative structure-activity relationships for hydrophobic interactions. Shirley. H. M. N. 1994. T. Suesa. 1991. Pharmacol. Relat. Kasuya. Z. Iwaki. W. P. Ogiso. [228] M. K. 34. H. R. Nissim. Duffield. Glycine and taurine conjugation of bile acids by a single enzyme. Guan. Kasumov. M. M. Hutt. New secondary metabolites of phenylbutyrate in humans and rats. 269. R. Tost. 239 – 249. [222] A. 31. 629 – 649. Pharm. Z. Fears. Nicholson. Cupid. 25. Garcia. M. D. Relationship to metabolic chiral inversion. 269. Okada. J. F. 1144 – 1149. Exp. [231] a) A. . Differences in the metabolism and pharmacokinetics of two structurally similar PPAR agonists in dogs: involvement of taurine conjugation. Drug Metab. T. Kochansky. Farrell. H. Comte. D. Xenobiotica 1999. Biochem. J. Dispos. Montserrat. 32. J. 153 – 163. Pharm. 1986. Tanino. S. Dispos. T. Kaiser. 194 – 200. Z. 847 – 854. X. S. Dispos. Moorhouse. [229] P. 2005. Pharmacol. G. Dispos. Xenobiotica 1990. K. Fujiwara. 1994. Vickery. Xenobiotica 2002. 17. R. Takagi. Lipid Res. K. Ortega. B. Gomez. E. B. Igarashi. L. Shirley. Day. Drug Metab. Kasuya. Absorption. 3403 – 3405. T. Acute dose-dependent disposition studies of nicotinic acid in rats. Comparative metabolism of high doses of aspirin in man and rat. Dodds. Ther. Biol. Dahms. Tang. Vabre. E. K. P. The stereoselective uptake of ibuprofen enantiomers into adipose tissue. Wang. B. Williams. I. 20. Kinman. 51. Participation of a medium chain acylCoA synthetase in glycine conjugation of the benzoic acid derivatives with the electron-donating groups. Bennett. K. D. 24. R. [221] M. David. Xenobiotica 1986. 10 – 19. Diasio. 2004. A. R. Thomas. Kim. 19375 – 19379. Pharmacol. b) M. S. Carabaza. C. 177 – 195. Taurine conjugation of ibuprofen in humans and in rat liver in vitro. K. Lipophilic xenobiotic conjugates: the pharmacological and toxicological consequences of the participation of drugs and other foreign compounds as substrates in lipid biosynthesis. A. 25. T. Masuda. T. A. b) K. R. Kanazu. [220] B. S. Barnes. Biol. C. D. Brunner. R. Struct. F. 27 – 42. Mis. 1991. 29. 5 (2008) 2325 [219] T. Stereoselective metabolic pathways of ketoprofen in the rat: incorporation into triacylglycerols and enantiomeric inversion. Kerr. F. Chirality 1996. Johnson. 41. 1997. Halstead. Comparison of in vitro carnitine and glycine conjugation with branched-side chain and cyclic side chain carboxylic acids in rats. R. 1994. Xenobiotic triacylglycerol formations in isolated hepatocytes. Smith. Bull. D. R. Patel. K. F. Igarashi. 16. S. Howell. Murphy. M. Wehrli. N. Puchowicz. R.CHEMISTRY & BIODIVERSITY – Vol. I. Gopaul. Z. L. Xenobiotica 2004. D. 1025 – 1042. B. S. M. A. A. 35. R. 1179 – 1185. Drug Metab. [225] C. M. Jobbins. 773 – 779. 34. The electronic effects of benzoic acid substituents on glycine conjugation. C. Stow. F. F. Wang. Dodds. 79 – 85. Biochem. Gutierrez. Res. Wilson. 24. F. Pharmacol. Hora. H. Lindon. K. D. T. T. Drug Metab. The metabolism of aspirin in man: a population study. [224] V. Baillie. 1996. L. Chem. N. G. Amino acid conjugates: metabolites of 2propylpentanoic acid (valproic acid) in epileptic patients. Fukui. J. Hardwick. T. Nanbo. Carganico. [226] G. D. Oxidative metabolism of a rexinoid and rapid phase II metabolite identification by mass spectrometry. Falany. 1986. Stereoselective taurine conjugation of (R)-benoxaprofen enantiomer in rats: in vivo and in vitro studies using rat hepatic mitochondria. Holmes. Vincent. 551 – 553. Incorporation of xenobiotic carboxylic acids into lipids by cultured 3T3-L1 adipocytes. W. Yamaguchi. K. J. Comparative molecular field analysis (CoMFA) including molecular lipophilicity potentials as applied to the glycine conjugation of aromatic as well as aliphatic carboxylic acids. Xenobiotica 2004. Hayashi. 1997. Prog. Benet. Lappin. 32. Hutson. L. D. 665 – 674. Jikihara. metabolism and excreation of 4-chloro-2-methylphenoxyacetic acid (MCPA) in rat and dog. P. J. 163 – 172. G. van Ravenzwaay. J. Biochem. Dodds. Lynn. Quantitative structuremetabolism relationships (QSMR) using computational chemistry: pattern recognition analysis and statistical prediction of phase II conjugation reactions of substituted benzoic acids in the rat. K. Knihinicki. [223] T. 15. X. L. G. Wheelan. 2003.-S. K. Pascual. 22. Incorporation of xenobiotic carboxylic acids into lipids. Brunengraber. Brunengraber. Abbott. 8. Benet. Dispos. L. 114 – 121. F. P. Caldwell. 149 – 153. H. Fukui. Quant.

157 – 169. hormone-sensitive and lipoprotein lipases. N. S. Dodds. Biol. Limited palmitoyl-CoA penetration into microsomal vesicles as evidenced by a highly latent ethanol acyltransferase activity. T. Murakami. 1998. 5 (2008) [232] G. 1255 – 1261. K. J. R. Xenobiotica 1982. Kaneko. F. D. Ito. 1991. Knights. 1995. following intravenous administration to the rat. 2000. [235] R. 521 – 525. Y.and sn-2-(3-phenoxybenzoyl)-dipalmitoylglycerol. [242] B. Ther. J. DiGiovanna. Matsubara. [243] G. 1986. P. D. Gantt. 663. A. Chem. 1599 – 1606. 1195 – 1201. Mizojiri. Baggaley. Exp. Bowden. Differential metabolism of fenvalerate and granuloma formation. L. p. Sanders. Melegh. 192 – 203. Yamaguchi. Saurat. A potentially new metabolic pathway: ethyl esterification of acitretin. 23. P. Okuno. Dispos. 324. Xenobiotica 1992. Schooley. Etretinate: persistent serum levels after long-term therapy. A. 1998. in Retinoids: 10 Years On. Woo. C. 83. L. Walker. Bell. Wiegand. R. In vitro studies to elucidate the metabolic pathway of (þ)-S-145. Drug metabolism in peroxisomes: involvement of peroxisomal b-oxidation system in the oxidative chain-shortening of xenobiotic acyl compounds. Xenobiotic cholesteryl ester formation. J. T. 1998. a novel thromboxane A2 receptor antagonist in rats. Schooley. J. A. Norikura. I. K. [239] K. M. 1986. Touchi. Hutson. rac-1. C. J. Dispos. The metabolism of 3-phenoxybenzoic acid-containing xenobiotic triacylglycerols in vitro by pancreatic. 1996. Biophys. Med. L. 462 – 464. P. In vitro metabolism of acitretin by human liver microsomes: evidence of an acitretinoyl-coenzyme A thioester conjugate in the transesterification to etretinate. Staiger. E. S. Karger. 83 – 89. Role and organization of peroxisomal b-oxidation. [246] Y. Siafaka-Kapadai. G. M. B. Ruddel. Baba. G. H. G. 125. J. Adv. Hindley. Pharmacol. [240] http://lpi. Biochim. [233] A. Van Veldhoven. P. Biochem. F. Wiegand. N. Huselton. L. Basel. [244] G. W. Kaneko. M. E. Pharmacol. Arch. Cell Biochem. Seki. Ulrich. T. 2008. K. 65 – 70. 26. K. Sherry. Casteels. Quistad. 1989. Matsuo. 1243. A. The metabolism of the xenobiotic triacylglycerols. M. Pharmacol. L. Pharmacokinet. Touchi. T. D. B. 613 – 626. Peck. A. A. Nakanishi. Javors. P.. L. Incorporation of [ 3H]valproic acid into lipids in GT1-7 neurons. B. Diep. U. Van Veldhoven. Hutson. Terracciano. Gasser. M. a thromboxane A2 receptor antagonist. Slatter. Tatsuhara. M. CHEMISTRY & BIODIVERSITY – Vol.. Dispos. R. Xenobiotica 1995. T. Drug Metab. T. Sägesser. Differential metabolism of fenvalerate and granuloma formation. N. T. H. Sugimoto. Nature 1982. M. 56.-H. Q. G. Yamaguchi. Acta 1995. H. T. 18. Suga. Zech. Murakami. Bøhmer. Reichen. 261 – 272. 1591 – 1598. Drug Metab. Sumegi. Bell. 296. [234] E. 246 – 251. Ed. G. 1986. Wyss. The role of carnitine in the conjugation of acidic xenobiotics. Biophys. . Yamada. Jensen. M. Biotransformation of lifibrol (U-83860) to mixed glyceride metabolites by rat and human hepatocytes in primary culture. Fears. T. Nishida. Dodds. B 1995. Pharmacokinetics of acitretin in humans. 207 – 212. D. J. Determination of urinary valproylcarnitine by GC-MS with selected-ion monitoring. [245] T.-W. Appl. Biol. Krähenbühl. H. Haselden. Watanabe. R. Miyamoto. 1999. 427 – 433. Quistad. 2000. 155 – 162. L. Biochem. Dermatol. R. Exp. Toxicol. P. C. Y. 83. M.. Staiger. D. H. Drug Metab. Mannaert. Sun. K. B. A. A. [238] U. T. 56. 73 – 87. Sex differences in the metabolism of (þ)-S-145. Biochem. N. Preferential elimination of pivalate with supplemented carnitine via formation of pivaloylcarnitine in man. J. Toxicity of valproic acid in mice with decreased plasma and tissue carnitine stores. [236] H. Xenobiotica 1993. S. F. Formation of pivaloylcarnitine in heart and brown adipose tissue in the rat. T.and alpha-oxidation in mammals. Drug Metab. 148 – 156. 60. Skrede. 568 – 575. Chromatogr. 12. 14. II. P. [241] H. J. Appl. Patiris. 993 – 1002. Peroxisomal lipid degradation via beta. Yamaguchi. Pharmacol.-W. 23. 2003. Xenobiotic conjugation: a novel role for bile acids. Toxicol. T. Y. E. [237] J. 221 – 231. Baba. Muro. Mannaerts. E. Yoshimori. Klemisch. Todesco. 56. J. P. A. Feenstra. P. Pharmacol. C. 507 – 516. 32. 24. P. Miyamoto. Biochem. 253. Beier. J. Chou. in rats. Matsubara. K. 22. Knapp. M. P. Pharmacol. M. 466. Polokoff. L. 7173 – 7178. R. R. 1978. Pharmacol.

H. R. Threadgill. R. Hiltunen. Miyazaki. K. Smith. Marsh. D. Kotti. D. pharmacokinetics and toxicity of ibuprofen enantiomers. Hall. J. T. B. 99. Garcia Alia. Interact. Interactions between the in vitro metabolism of xenobiotics and fatty acids. Weinstein. Pharmacol. Houghton. Horner. Helander. T. 215 – 234. Sormunen. J. Acyl-coenzyme A formation of simastatin in mouse liver preparations. 1993. Wanders. Q.7a. Hao. A. 1992. 275. Hutt. K. Arano. Denis. G. S. I. J. G. Moss. B. A. H. H. R. Savolainen. Pharmacol. Drug Metab. Pharmacodynamics. A. W. Pharmacol. Y. T. Testa. Dispos. Marsh. 2008. W. [250] M. Biochem. G. C. Technetium-99m-labelled long chain fatty acid analogues metabolized by b-oxidation in the heart. Irie. E. Expression of rat liver long-chain acyl-CoA synthetase and characterization of its role in the metabolism of R-ibuprofen and other fatty acid-like xenobiotics. Drug Metab. K. Eichelbaum. M. Dispos. Ferdinandusse. Mayer. V. 1982. E. Biomed. T. H. Miyano.and 3-keto-3-phenylpropionic acids: novel metabolites of benzoic acid in horse urine. Rev. 2. J. 2003. Enantioselective pharmacokinetics of ibuprofen and involved mechanisms. Hunninghake. L. Xiaotao. P. 499 – 513. [255] B. Didonato. Moss. L. T. Yamanaka. Ishibashi. Tegeder. B. Sun. 34. Ikegawa. Woolf. Mizugaki. J. E. Y. S. 2000. Owatanabe. 1986. M. 341 – 354. P. H.2-dimethyl-N-(2. 1890 – 1896. A. A novel metabolic pathway in the metabolism of 5-(4’-chloro-n-butyl)picolinic acid. D. Biochem. Till. Testa. Takayama. Holmas. Molecular cloning of cDNA species for rat and mouse liver a-methylacyl-CoA racemases. F. Yamamoto. E. B. Uehara. Cuebas. Metabolic conjugation of some carboxylic acids in the horse. W. [252] I. p. Yu. 883 – 889. 2001. L. Testa. Phillips. R. J. Chirality 1995. J. Dispos. in Stereochemical Aspects of Drug Action and Disposition. Eds. H. Ulm. Biotransformation of lovastin. D. 235 – 251. 740 – 748. Houghton. M. Suppl. in Stereochemical Aspects of Drug Action and Disposition. H. M. J. 42. 7.and (25S)-3a. Audergon. M. 3225 – 3230. B. 41. A. Williams. S. Brune. Savolainen. T. I. 1347 – 1366. The role of coenzyme A in the biotransformation of 2-arylpropionic acids. E. J. J.CHEMISTRY & BIODIVERSITY – Vol. Darley. E. Miyazaki. Sagi. M. [253] H. S. 102 – 110. W. Eur. Y. Minatogawa. 1997. K. A mouse model for alpha-methylacyl-CoA racemase deficiency: . S. Y. S. Schmitz. Duggan. A. Drug Metab. Benoit. Springer Verlag. K. Subramanian. J. M. 61. Pan. Mayer. Chando. D. M. Metabolic chiral inversion of 2-arylpropionic acids. 3. J. Soraci. V. 50. 37. A. Prueksaritanont. Black. H. 2005. Akizawa. Novikov. Testa. Vyas. 1991. Drugs Future 1997. M. FEBS J. 801 – 807. T. R. Mass Spectrom.4. D. 469 – 476. E. Drug Metab. 1995. Eds. J. Med. J. S. S. R. 19. Dacremont. M. Goto. 21. Waterham. p. M. M. M. Brugger. S. Halpin. Springer Verlag. J. Lloyd. Berlin. M. Smith. Enantiomer 1997. M. A. G. 543 – 549. Biotransformation of pravastatin sodium in humans. Arch. Schmitz. J. [256] W. [251] H. H. C. Goto. 25. H.-Biol. T. Geisslinger. St. Wang. W. Hiltunen. 2002. K. Caldwell. C. Li. Testa. 2007. 363. J. Somogyi. 2003. Stereoisomeric bio-inversion of (25R). 123 – 130. Hirabayashi. D. S. 1089 – 1102. M. B. R. E. E. Ilves. H. W. Somogyi. Alpha-methylacyl-CoA racemase – an obscure metabolic enzyme takes centre stage. K. Conzelmann. M. K. Takayama. Odaka. M. 22. Mayer. 5 (2008) 2327 [247] T. Biochem. 2006. K. Sinz. Chiral recognition in biochemical pharmacology: An overview. The case of ibuprofen and other chiral profens. 90. Lipid Res. Uemura.12atrihydroxy-5b-cholestanoic acid CoA thioesters in rat liver peroxisome. Chem. Clin. 1976. Singhvi. Vainio. Toxicol. 1994. [249] C. J. 143 – 159. K. C. 333 – 342. T. J. M. 1997. Enantioselective pharmacodynamics and pharmacokinetics of chiral non-steroidal anti-inflammatory drugs. Berlin. M. Xenobiotica 1981. D. Drug Metab. S. 140 – 145. S. 651 – 656. J. [254] R. Dispos. Trivedi. A. In vitro and in vivo disposition of 2. Roy-de Vos. Reichel. Kari. Tegeder. Caldwell. T. Subcellular localization and physiological role of alpha-methylacyl-CoA racemase. Chem. Conzelmann. Schmitz. 326. Conzelmann. 655 – 663. 11. Everett. A. M. 17. Chain elongation of fusaric acid and related compounds in rat liver. S. Biochem. Evans. Wierzbicki. A. 1003 – 1011. Biochem. G. Geisslinger. J. 31. 3Hydroxy. E. The role of alphamethylacyl-CoA racemase in bile acid synthesis. 534 – 540. Bjorge. Adachi. H. Horner. [248] M. Magata. Eichelbaum. B. In vitro fenoprofenyl-coenzyme A thioester formation: interspecies variation. 237 – 256. IJlst. [257] D.6-trimethoxyphenyl)dodecanamide (CI-976).

Kohno. J. J. J. Ray. B. Influence of age on the enantiomeric disposition of ibuprofen in healthy volunteers. Biochem. Ho. The oxidizing power of the glutathione thiyl radical as measured by its electrode potential at physiological pH. E. Duff. A. Glutathione and its role in cellular functions. Pharm. Glutathione metabolism and its role in hepatotoxicity. Biophys. Pharmacol. Meister. Rogers. D. 1996. Ross. Kurtz. 2004. Lin. P. Green. J. and diabetes. 74. Kishimoto. R. 1980. S. S. T. Hutt. Varnes. Jackson. Madej. Wolen. 2007. Pharm. 5 (2008) [258] [259] [260] [261] [262] [263] [264] [265] [266] [267] [268] adjustement of bile acid synthesis and intolerance to dietary methyl-branched lipids. Casini. Pharmacokinetics and bioinversion of ibuprofen enantiomers in humans. Visvikis. Clin. Glutathione in human plasma: decline in associaiton with aging. Drug Metab. Pharmacol. J. M. C. D. 34. J. 36. Mol. Millis. Andersen. Grubb. D. T. Woodage. E. J. H. Samiec. M. R. 1037 – 1048. Rev. 4144 – 4146. P. K. J. 383 – 387. S. 1499 – 1503. A. 1 – 12. N. V. Lipid Res.and beta-oxidation systems in patients suffering from different peroxisomal disorders. Rajagopalan. Drug Metab. Br. D. Forman. H. Free Radical Biol.2328 CHEMISTRY & BIODIVERSITY – Vol. Weaver. Biochem. Drews-Botsch. 821 – 863. D. Rev. P. J. Clin. 52. L. P. 64. Swift. 58. glutathione. G. Cellular glutathione and thiols metabolism. Y. D. Effect of absorption rate on pharmacokinetics of ibuprofen in relation to chiral inversion in humans. E. F. C. E. 66. Hum. E. Reed. K. De Tata. in rat. E. S. Y. Bharath. Jones. 169 – 172. L. takeuchi. R. Hayakawa. Biochem. Stereospecific inversion of (R)(À)-benoxaprofen in rat and man.. S. (S)-ketoprofen in eight animal species. Brater. H. S. Stereoselectivity of ibuprofen metabolism and pharmacokinetics following the administration of the racemate to healthy volunteers. E. Epp. Wardman. N. A. 42. Methods Enzymol. S. R. Rubin. J. Rao. Drug Metab. Holland. C. Hutt. Arch. D. D. J. Glutathione. C. 1985. Calculation of inversion half-lives of arylpropionic acid class nonsteroidal antiinflammatory drugs. Biaglow. W. Mechanistic studies on metabolic chiral inversion of 4-(4-methylphenyl)-2-methylthiomethyl-4-oxobutanoic acid (KE-748). J. Inotsume. A. Xenobiotica 2002. Mason. a cellular protagonist. Pharm. 17205 – 17208. Chem. P. 55. K. G. Jackson. Fukushima. Stereochemistry of the peroxisomal branched-chain fatty acid alpha. 1990. 1999. B. Ding. Held. Genet. H. 955 – 965. G. 1989. Chem. 2002. K. D. Aberg. K. K. Sci. Rep. H. Ther. Small-molecule inhibitors of glutathione S-transferase P1-1 as anticancer therapeutic agents. Ciofalo. M. Org. 318 – 329. J. Pendleton. Role of glutathione and other thiols in cellular reponse to radiation and drugs. T. V. R. J. J. Endo. DeLeve. A. 1991. 1999. D. age-related macular degeneration. Mulder. B. H. Biochem. Sternberg Jr. A. J. 5. 20. Suwa. Yamaguchi. J. Demetriades. Med. Stereoselective inversion of (R)-fenoprofen to (S)-fenoprofen in humans. 1509 – 1513. G. H. D. 824 – 830. 438 – 444. 1993. 287 – 307. Burg. an active metabolite of the new anti-rheumatic agent 2-acetylthiomethyl-4-(4methylphenyl)-4-oxobutanoic acid (KE-298). and proteins. A. Clin. L. J. R. A. Pharmacol. D. R. N. 321 – 335. 2002. 82 – 84. R. Tuttle. Res. N. Stereoselective pharmacokinetics of ketoprofen and ketoprofen glucuronide in end-stage renal disease: evidence for a futile cycle of elimination. W. H. Rudy. 1997. S. P. Knadler. D. L. G. 2003. S. N. Toda. Pharmacol. R. C. G. Oxidation/reduction potential of glutathione. Fukushima. A. Swift. D. The changing faces of glutathione. L. 43. 2003. Inversion of (R). G. 2002. M. Depuy. T. Morales. Free Radical Biol. Med. 24. 494 – 500. L. Rabenstein. Br. D. van Lint. Tan. Laborde. 1998. J. S. Kantoci. Dickinson. Pompella. Vreken. Sun. Pharmacol. G. P. G. Kaur. Simmonds. W. 179 – 187. 7. S. Pharmacol. A. 500 – 504. P. 2007. Seibold. . Pharmacokinet. 1019 – 1026. 2002. Clark. 48. S. Rusch. 1988. C. Wanders. Kaplowitz. E. S. 462. Hall. Eur. J. Paolicchi. Yoshida. Liu. 59. Patel. Flagg. G. Patel. J. 13. J. Chem. 64. M. 1994. J. Weddle. P. 53. G. Wechter. Biol. D. Pharmacol. iron and Parkinsons disease. Ferdinandusse. C. 2007. 683 – 697. Tan. 579 – 587. Glutathione conjugates and their synthetic derivatives as inhibitors of glutathione-dependent enzymes involved in cancer and drug resistance. Pharmacol. Glutathione metabolism and its selective modification. J. S. Thiyl free radical metabolites of thiol drugs. Hayashi. H. K. 94 – 102. Y. 186. 283. P. Pharmacol. Bechtol. Dacremont. D. W. W. Chirality 1995. 27. Annu. A. Hsu. Sies. 11. Biochem. Cheng. E. 916 – 921. 32. K. 699 – 704. J.

G. Elcombe. G. Res. Hayes. Schnekenburger. Med. A. Drug Metab. M. Thomson. 22. Schaid. D. P. Structure. R. The three-dimensional structure of a class-Pi glutathione S-transferase complexed with glutathione: the active-site hydration provides insights into the reaction mechanism. N. Effect of vitamin E on glutathione-dependent enzymes. E. 30. E. Rev. 10. Rev. Biochem. M. Commandeur. 45. Biochem. J. S. H. 2001. J. Chem. Murdock. J. Mitchell. Diederich. J. V. Robinson. L. A. Tzeng. N. T. van der Aar. Modelling and bioinformatics studies of the human kappa-class glutathione transferase predict a novel third glutathione transferase family with similarity to prokaryotic 2-hydroxy-2-carboxylate isomerases. Dowd. Metodiewa. C. T. Dispos. Biochem. Biochem. Morceau. I. Takahashi. Structure. Anders. D. K. C. Biophys. E.-H. D. 68. A. 1988. 1992. T. S. 445 – 600. M. Jowsey. N. Sheenan. Tyrosine-7 in human class Pi glutathione Stransferase is important for lowering the pKa of the thiol group of glutathione in the enzymeglutathione complex. 271 – 276. Fahmy. Booth. Toxicol. Tsuchida. 1997. Eckloff. 559 – 569. 34. P. 569 – 643. Walsh. 39. Zhang. Estrella. 231 – 249. Biochemical and genetic characterization of a murine class kappa glutathione S-transferase. Biochem. Evelo. J. Tsuchida. Mol.-F. Annu. Biochem. M. Parraga. 51 – 88. Huttley. G. Hayes. Glutathione transferases. R. Pharmacol. P. R. E. Biochem. M. M. 44. Toxicol. M. Pelliger. C. Meade. Crit. Biophys. Arch. Winterbourn. Weinshilboum. Armstrong. M. 1999. C. A. 1995. Ther. Drug Metab. P. H. Mammalian class sigma glutathione S-transferases: catalytic properties and tissue specific expression of human and rat GSH-dependent prostaglandin D2 synthases. 284 – 290. J. J. Identification of novel glutathione transferases and polymorphic variants by expressed sequence tag database analysis. 4296 – 4299. Biochem. S. T. J. free radicals and chemotherapeutic agents – Mechanisms of free-radical induced toxicity and glutathione-dependent protection. N. 1905 – 1915. and evolution of the glutathione transferases. J. C. C. Bast. Protective effect of N-acetylcysteine against radiation induced DNA damage and hepatic toxicity in rats. Pharmacol. Haenen. R. C. S. R. M. Delhalle. H. Contribution of tyrosine 6 to the catalytic mechanism of isoenzyme 3-3 of glutathione S-transferase. 269 – 274. Pharmacol. 373. A. . Biochem. 75. 1992. D. 2001. G. Biochem. 29. Glutathione transferases and cancer. 35. X. function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. Flanagan. Biochem. L. Coll. 1992. 541 – 552. L. 379. Drug Metab. 314. K. 1988. 2005. Mol. Biol. B. Sato. P. D. M. 1998. The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. M. Kong. D. R. C. a) R. M. 5 (2008) 2329 [269] [270] [271] [272] [273] [274] [275] [276] [277] [278] Glutathione. Meyer. B. Jowsey. Rev. Blood glutathione as an index of radiation-induced oxidative stress in mice and humans. G. Increased glutathione S-transferase P1-1 expression by mRNA stabilization in hemin-indiced differentiation of K562 cells. M. M. J. R. D. Free Radical Biol. Pulford. Murphy. Pharmacol. Hayes. A. K. Rev. Suzuki. Trentesaux. van Haaften. Flanagan. Mukherjee. 1269 – 1277. Role of glutathione and other endogenous thiols in radiation protection. Biol. 2003. J. W. F. Strategies to characterize the mechanisms of action of glutathione S-transferases: a review. 1997. Navarro. E. Ji. K. Moore. J. 2006. Biophys. Smith. M. Satoh. B. 360. Glutathione S-transferase omega 1 and omega 2 pharmacogenetics. Moon. Vermeulen. 359. Drug Metab. 1994. U. 1998. Sato. 773 – 780. A. S. H. Hayakari. Arch. Obrador. 1 – 16. I. Biochem. Board. 27. 811 – 816. D. 1203 – 1209. Res. H. Duvoix. 333. 2003. 2004. Tan. Pharmacol. H. Blackburn. T. A. Vina. 544 – 547. Armstrong. S. Asensi. 215 – 253. Gilliland. Wieben. R. Foley. B. A. N. N. F. 30. N. Jowsey. On the antioxidant effects of ascorbic acid and glutathione. J. J. K. 2008. I. U. 2 – 18. O. M. 2001. M. Orton. Liu. Rev. D. Ther. R. 194 – 197. b) E. L. Jermin. A. R. Mansour. Chelvanayagam. Hatayama. J. W. Pharmacol. 267. I. Takasu. W. E. G. The reaction of superoxide with reduced glutathione. A.CHEMISTRY & BIODIVERSITY – Vol. Salavaggione. A. Hayes. 361. Board. W. 507 – 516. Mantle. K. A. Crit. Dispos. R. S. S. G. Biaglow. I. Quantitative differences in the active-site hydrophobicity of five human glutathione S-transferases. D. J. Meister. J. Biochem. 1992. G. Thomson. 1237 – 1246. Takahata. Pelleymounter. Garcia-Saez. Commun. 2004. B. J. catalytic mechanism. Hanafi. Johnson. J. Dicato. J. Chem. T. G. 337 – 384. J. S. 37. Biol. J. Russo. J. Inoue. I. Tetlow. 184. J. Hafez. I.

Med. c) B. Interact. [282] J. S. Ed. S. b) R. 1982. 265 – 270. Adachi. T. E. S. Rev. B. 2006. Z. MRP class of human ATP binding cassette (ABC) transporters: historical background and new research directions. 271 – 330. 22179 – 22185. Xenobiotica 1992. M. R. Matsumoto. G. Villalta. 351 – 360. Glutathione and the response of malignant cells to chemotherapy. p.10-dihydro-9-hydroxy-10-phenanthryl)-d-cysteine in human urine: comparison with glutathione-S-transferase genotypes in smokers. Chem. Lindstrom. Bondar. Ballatori. S. 18. [294] A. Trends Pharmacol. H. Dekant. B. 2005. Chem. E. Dispos. Mulder. 113 – 121. Nuccetelli. 35. Drug Metab. P. S. Ketterer. Drug Metab. P. . Chem. Simmons. [283] a) N. Scholz. 1053 – 1058. G. 43 – 50. Res. K. 5 (2008) [279] A. Drug Metab. Quantification of N-acetylS-(9. in Conjugation Reactions in Drug Metabolism. Ryazantsev. [289] C. Sies. Res. Res. Environ. Chem. Hamill. Stijntjes. Vermeulen. Gillam. Xenobiotica 1993. S. 455 – 464. G. H. Watson. 56. 1996. 105. J. J. W. W. Commandeur. Rinaldi. Ketterer. 271. 16193 – 16198. Kearns. Kinetics of the conjugation of aniline mustards with glutathione and thiosulfate. Clegg. 1135 – 1145. Rev. 1009 – 1014. T. Toxicol. Drug Metab. Bleasdale. Detoxication reactions of glutathione and glutathione transferases. Toxicol. L. V. W. J. W. Dekant. 1984. 1991. 38. J. Masubuchi. C. P. 1992. R. Akerboom. A. Reactions of benzene oxide with thiols including glutathione. Chem. Intrahepatic conversion of a glutathione conjugate to its mercapturic acid. P. 1234 – 1240. N. D. P. 2007. 1982. M. P. [281] a) B. Specificity of aminoacylase III-mediated deacetylation of mercapturic acids. Dirven. Kurtz. Ketterer. 3. The role of nonenzymatic reactions of glutathione in xenobiotic metabolism. 11. 1996. J. 2001. Sassani. London. 30. Barnes. Li. J. 161 – 187. M. W. The role of human glutathione transferases and epoxide hydrolases in the metabolism of xenobiotics. Antonini. 20. Makino. Vamvakas. Morgenstern. R. Hashmi. Glutathione-dependent bioactivation of xenobiotics. Anders. [295] P. M. Eliasson. Xenobiotica 1986. P. Rappaport. Ishikawa. W. Ascenzi. Kuo. A. J. G. T. T. 22. Cashman. Upadhyaya. 2002. [286] Y. S. Rev. B. M. Hochalter. Colvin. Bellizia. Health Perspect. Millis. N. M. A. [291] S. Free Radical Biol. [288] C. M. 16. Res. NMR studies of the conjugation of mechlorethamine with glutathione. A. Murayama. J. Oakley. 1307 – 1334. Sci. [285] J. Flavincontaining monooxygenase-dependent stereoselective S-oxygenation and cytotoxicity of cysteine-Sconjugates and mercapturates. R. P. 1995. 5. Enzymes and transport systems involved in the formation and disposition of glutathione S-conjugates. Newman. Parker. N. N. Dispos. J. 307 – 364. Cameron. Rao. 791 – 799. N. B.-Biol. Toxicol. Chem. Toyoda. Prediction of in vivo potential for metabolic activation of drugs into chemically reactive intermediates: correlation of in vitro and in vivo generation of reactive intermediates and in vitro glutathione conjugate formation in rats and humans. 9. A. M. A. Drug Metab. [292] M. Hall. T. Glutathione conjugation. G. Chem. S. 957 – 973. Gramcsik. E. Henderson. K. S. Taylor & Francis. Pharmacol. Biochem. Pharmacol. Swedmark. 15. Nonenzymatic contributions to xenobiotic metabolism. Chem. Glutathione-dependent toxicity. Tsuprun. [287] P. Med. M. Hinchman. G. J. van Ommen. J. W. I. Keppler. 35 – 52. G. 47. Testa. T. G. P. M. Seidegard. G. 1999. Vamvakas. van Bladeren. 105. Abuladze. D. 873 – 887. Hagiya. Export pumps for glutathione S-conjugates. P. Xenobiotica 2008. A. 763 – 771. Biol. Gramcsik. K. Battistoni. Waidyanatha. Coles. Dekant. M. Y. 833 – 862. b) B. Rev. 2007. Golding. Toxicol. Hecht. 1990. Structural flexibility modulates the activity of human glutathione transferase P1-1. Heath. Osterloh. Biol. Pushkin. 193 – 201. Ekstrçm. Glutathione conjugation of alkylating cytostatic drugs with a nitrogen mustard group and the role of glutathione S-transferases. 19. Toxicol. T. M. 13. Mulder. Caccuri. G. Reactive intermediates and the dynamics of glutathione transferases. W.  Hepatic mercapturic acid formation: involvement of cytosolic cysteinylglycine S-conjugate dipeptidase activity. Chem Res. [284] E. Effects of modifying structure on electrophilic reactions with biological nucleophiles. L. 22. 1997. 1998.2330 CHEMISTRY & BIODIVERSITY – Vol. W. [280] a) B. The dark side of a detoxification mechanism. 25 – 50. K. C. 1990. H. b) B. 33. 985 – 991. 13. W. T. A. M. Ricci. M. Jçsch. [293] M. Vamvakas. S. J. 1997. B. [290] D. Park. 266. 27. Hamill. Anders. P. 23. A. Drug Discovery Today 1998. Chiessi. Hoshijima.

E. M. 1996. U. K. K. Chem. S. Srivastava. S. Stereochemistry of styrene biotransformation. [301] J. 430 – 436. Portoghese. Reactivity of glutathione with a. Zjang. Berhane. Structural characterizaiton of in vivo rat glutathione adducts and a hydroxylated metabolite of simvastatin hydroxy acid. 580 – 585. 2001. 2001. A. Res. [297] J. Williams. A. Lau. 2002. Scobie. Free Radical Biol. Chen. Toxicol. [311] F. S. E. 1989. [299] J. 1829 – 1841. F. Chem. Dispos. Dispos. Drug Metab. Elfarra. 1991. Breau. Baillie. 2001. L. 1085 – 1093. 8. [304] S. Drug Metab. 73. 31. Mannervik. Drug Metab. Rev. and rearrangements of the glutathione conjugates of butadiene monoxide: evidence for the formation of stable sulfurane intermediates. Y.S. C. H. K. [307] D. Food Chem. 1994. S. Eklund. H. a novel metabolite of morphine from guinea pig bile. Toxicol. Fujioka. Mannervik. R. 1999. J. Vodicka. Verlukast (MK-0679) conjugation with glutathione by rat liver and kidney cytosols and excretion in the bile. Twa. Ito. Interactions of glutathione Stransferase-p with ethacrynic acid and its glutathione conjugate. Acad.  Human glutathione transferases catalyzing the bioactivation of anticancer thiopurine prodrugs. 2006. G. J. Hachiyama. hydrolytic. A. [312] M. 1994. 773 – 776. [310] M. J. chlorogenic acid. Stereoselective catalysis of a retro-Michael reaction by class mu glutathione transferases. J. Kemper. R. Vodickova. P. T. Engstrçm. Krause. S. B. [302] T. Yano. 1480 – 1484. L. P. Mihashi. Acta 1993. K. 27. 23. Trimble. Elfarra. A. [298] I. Bible Jr. Youngman. D. Subramanian. 1989. 77 – 81. Murphy. T. (8S)-(Glutathion-S-yl)dihydromorphinone. H. Selvaraj. [303] a) K. Ishida. S. Moridani. Quinones in biology: functions in electron transfer and oxygen activation. X.A. Armstrong. A. L. F. A. P. Dispos. 1432 – 1439. Proc. 1152 – 1160. Widersten. valdecoxib. Toxicol. 211 – 279. Dispos. Peters. J. T. P. 2. W. S. Salehi. Monks. Ansari.. A. Res. Monks. 2007. Larson. Boyland. 5 (2008) 2331 [296] R. Lau. R. D. Fang. Duescher. Š. S. Elfarra. L. Toxicol. Y. M. Chem. Kumagai. B. Metabolism of tert-butylhydroquinone to S-substituted conjugates in the male Fischer 344 rat. Biochem. and potential carcinogenicity. Urinary metabolites from F344 rats and B6C3F1 mice coadministered acrylamide and acrylonitrile for 1 or 5 days. Ahmad. 2003. 1967. J. Res. K. Detoxication of base propenals and other a. G. Gunnarsdottir. Chasseaud. Drug Metab. Drug Metab. [305] T. Biochem. J. A. 225 – 230. Sumner. 805 – 853. [309] H. Formation. van Bladeren. .b-unsaturated ketone flavouring substances. Y. B. Kozarich. N.CHEMISTRY & BIODIVERSITY – Vol. A. 1997. 38. Toki. J. T. D. Jordan. Drug Metab. Y. G. J. Hemminki. in mice. [306] S. 4. Yuan. Y. R. Yergey. Linhart. Oesch-Bartlomowicz. [308] B. Hall. A. 1164. Wang. Awasthi. K. Y. Kedziora. J. S. Kassahun. Jamshidzadeh. R. Biophys. Nicholl-Griffith. Res. 104. Chem. Dispos. B. Natl. 29. Rev. Glutathione and N-acetylcysteine conjugates of a-methyldopamine produce serotonergic neurotoxicity: possible role in methylenedioxyamphetamine-mediated neurotoxicity. N. 12. 830 – 836. F. Metabolism of butadiene monoxide by freshly isolated hepatocytes from mice and rats: different partitioning between oxidative. L. Med. S. Drug Metab. Gupta. van Ommen. A. Ikeda. and dihydrocaffeic acid metabolism: glutathione conjugate formation. 1995. Bernard. Sci. Adv. C. Nohl. Prueksaritanont. Iwase. T. L. Biochim. R. M. Drug Metab. 29. Dispos. 1995. A. 95 – 102. Consequences for the internal distribution of products in the active site. Pharmacol. and conjugation pathways. 353 – 367. J. Naccarati. Toxicol. genotoxicity. I. Styrene metabolism. A. c) E. W. Chem. Bai. Toxicol. 17. S. S. 10. OBrien. Caffeic acid. M. Nauhaus. K. 491 – 501. 91. 1150 – 1157. Pharmacol. Oesch. P. Ahmad. Reversibility in glutathione-conjugate formation. Sharer. J. H. Fennell. stability. P. 163 – 181. 33. Res. 173 – 178. Adv. B. 133 – 139. 30.b-unsaturated aldehyde products of radical reactions and lipid peroxidation by human glutathione transferases. S. R. Enzyme-catalysed conjugations of glutathione with unsaturated compounds. Pharmacokinetics and metabolism of a COX-2 inhibitor. Dulik. 1986. F. [300] R. 27. 9. Koskinen. b) P. A. J.

229 – 255. 709 – 717. Biochem. C. M. van Bladeren. M. Rev. T. Fundam. The regioselectivity of glutathione adduct formation with flavonoid quinone/quinone methides is pHdependent. Use of 19FNMR and GC-electron capture detection in the quantitative analysis of fluorine-containing metabolites in urine of sevoflurane-anesthetized patients. K. J. Pharmacol. B. M.-Biol. B. Structure-activity relationship for the biotransformation of haloalkenes by rat liver microsomal glutathione transferase I. M. Anders. Chem. L. Appl. D. 118. Macdonald. B. Toxicol. 15. 1997. M. Baggs. J. [325] S.2-difluoro-1-(trifluoromethyl)vinyl ether (compound A). [316] J. [315] D. Iverson. Chem. Nephrotoxicity of the glutathione and cysteine S-conjugates of the sevoflurane degradation product 2-(fluoromethoxy)-1. Res. R. P. Kanhai. Boersma. M. E. C. 261. Ther. Toxicol. Methyl isocyanate: a review of health effects research since Bhopal. S. M. 2003. Szymusiak. 1036 – 1041. R. Boersma. Formation of glutathione conjugates during oxidation of eugenol by microsomal fractions of rat liver and lung. Deese. ´ 666 – 668. Anders. Drug Metab. [321] L. Lau. M. 10. Sahin. 13. Park. E. 129 – 162. Reactions of quinone methides with proteins: analysis of myoglobin adduct formation by electrospray mass spectrometry. Vervoort. S. 15. S. Kharasch. I. Toxicol. Commandeur.2332 CHEMISTRY & BIODIVERSITY – Vol. 1993. Chem. 1248 – 1252. 1302 – 1308. Res. Bucher. Pharmacol. Sulfoxidation of cysteine and mercapturic acid conjugates of the sevoflurane degradation product fluoromethyl-2. ´ [317] D. Vervoort. M. 1296 – 1313. Chem. Bolton. 21. D. P. E. 1996. M. P. J. Thompson. Bowlin. B. 1989. Ther. M. J. De Kanter. J. 1998. 2001. 367 – 379. J. Toxicol. Anders. 36. S. Smith. P. J. Annu. Fitzsimmons. [322] M. B. 1992. U. Henschler. J. M. Dispos. Harman. Xenobiotica 2004. D. J.3. 17. Drug Metab. E. 1987. 583 – 594. 717 – 721. . W. A. 2004. Toxicol. Dalvie. W. Rev. N. Trugman. B. Am. Moldeus. Finkelstein. W. 30. S. Chem. 34. Biol. Annu. 343 – 351. G. Aypar. Cenas. Baggs. Stereochemistry of the microsomal glutathione S-transferase catalyzed addition of glutathione to chlorotrifluoroethene. L. 905 – 916. W. Commandeur. M. P. Toxicol. Egestad. Res. Lemanska. J. I. Exp. Pharmacol. Y. Pharmacol. F. N. S. Identification of a reactive metabolite of terbinafine: insights into terbinafine-induced hepatotoxicity. J. The pharmacology and toxicology of polyphenolic-glutathione conjugates. W. Toxicol. [324] W. Iyer. [314] K. N. [326] J. Toxicol. W. Sugumaran. D. Toxicol. N. Tyrakowska. Res.3. 22. J. 38. Nephrotoxicity of the glutathione and cysteine conjugates of 2-bromo-2-chloro-1. 2004. 1997. W. 2002. M. 14. 38. Exp. H. Mol. 39. M. 301 – 316. Uetrecht. A. C. G. J. 1992. Chem. Biological and toxicological consequences of quinone methide formation. Segura-Aguilar. G. Smith. Chem. Aniya. R. Res. Moldeus. Anders. Biochemistry 1991. N. 34. J. W. G. P. 435 – 445. Glutathione-dependent bioactivation of haloalkanes and haloalkenes. Reactions of glutathione with carcinogenic esters of N-arylhydroxamic acids. Koob. Anders. 1990.1-difluoroethene. Thompson. Chem. Siu. 1544 – 1551. Thompson. In vitro metabolic activation of thiabendazole via 5hydroxythiabendazole: identification of a glutathione conjugate of 5-hydroxythiabendazole. Res. R. S. Mass Spectrom. J. L. Hargus. Y. M. J. Lin. Chem. M. Le Blanc. Pharmacol. 86. Jolivette. [320] M. S.1. Smith. Novak. Glutathione-dependent bioactivation of haloalkenes. Altuntas. In vitro metabolism of tolcapone to reactive intermediates: relevance to tolcapone liver toxicity. Soc. D. H. 16. 1587 – 1595. 654 – 663. 2000. 175 – 181. J. Boeren. 123 – 128.3-pentafluoro-1-propene (compound A) in male Fischer 344 rats. C. T. J. T. 283. Interact. 2006. 1998. T. 5 (2008) [313] T. Rietjens. [323] H. K. 36. Toxicol. S. 9. Mickos. [318] H. Regioselectivity and reversibility of the glutathione conjugation of quercetin quinone methide. Rietjens. Anders. M. Heady. W. Res. 2002. A. P. A. 185 – 191. Biological reactivity of polyphenolic-glutathione conjugates. Vermeulen. L. J. Pharmacol. W. Sahin. Monks. [319] N. Bioactivation of the cysteine-Sconjugate and mercapturic acid of tetrafluoroethylene to acylating reactive intermediates in the rat: dependence of activation and deactivation activities on acetyl coenzyme A availability. N. Dekant. Vermeulen. Lau. 501 – 537. Xenobiotica 1991. M. Rev. Metabolism of 14C-dichloroethyne in rats. Monks. Orhan. Constantin-Teodosiu. J. Awad. E. Dekant.

1107 – 1117. Snel. K. N. 2003. J. J. Wheeler. 2001. Stourman. 12. Kim. 1118 – 1127. 264 – 270. M. S. Res. McCormick. T. D. Characterization of novel isocyanate-derived metabolites of the formamide N-formylamphetamine with the combined use of electrospray MS and stable isotope methodology. R. Asmuth. Toxicol. Armstrong. Ehlhardt. Arch. 14. F. Oda. Dommermuth. Botta. T. 1091 – 1096. C. 2001. K. Toxicol. Kagan. S. Y. R. Mundkowski. Glutathione transferase theta 1-1-dependent metabolism of the water disinfection byproduct bromodichloromethane. Pearson. Toxicol. K. Res. P. Chem. Chem. Jeong. W. 891 – 899. Baillie. Abbott. B. 240 – 248. W. R. W. 600 – 608. G. Shimada. 4. Schrçder. M. Baillie. B. D. T. 17. M. Chem. and human liver cytosol: 13C NMR spectroscopic studies. Watanabe. Lange. W. M. Borel. Chem. A. Toxicol. Chem. Dechert. Chem. Toxicol. Toxicol. Guengerich. T. 14. Biochem. 24. G. Conjugation of haloalkanes by bacterial and mammalian glutathione transferases: mono. P. 1228 – 1233. E. Wheeler. Res. [336] K. 68. Rashed. W. Watabe. [333] D. Biotransformation of the naturally occurring isothiocyanate sulforaphane in the rat: identification of phase I metabolites and glutathione conjugates. W. Conjugation of haloalkanes by bacterial and mammalian glutathione transferases: mono. K. Res. L. 2002. A. C. Baillie. Baillie. R. Guengerich. A. 291 – 296. Dommermuth. 15. Meyer. Kolm. Guengerich. J. Lee. Slatter. V. Glutathione-dependent metabolism of the antitumor agent sulofenur. Talalay. Biotransformation of methyl isocyanate in the rat. A. J. H. Kassahun. [337] M. F. 311. Toxicol. Glutathione conjugation and pharmacokinetics of 2-bromo-3-phenylpropionic acid in vitro and in the rat in vivo. 16. 4. A. Guengerich. Res. Res. Jeong. Stourman. E. 423 – 427. H. P. A. Morris. Slatter. Toxicol. Toxicol. V. N. Hyun. Yamazaki. 5 (2008) 2333 [327] J. P. Characterization of nucleoside and DNA adducts formed by S-(1-acetoxymethyl)glutathione and implications for dihalomethane-glutathione conjugates. 1994. Res. M. M. 1991. Identification of glutathione conjugates and mercapturic acids of 1. [332] K. rat. M. A. Mulder. J. Analysis of the kinetic mechanism of haloalkane conjugation by mammalian theta-class glutathione transferases. Manier. T. G. Jin.and vicinal dihaloethanes. S. K. F. P. 407 – 414. Toxicol. R. Lee. Intracellular Sglutathionyl adducts in murine lung and human bronchoepithelial cells after exposure to diisocyanatotoluene. Y.2dibromopropane in female BALB/c mice by liquid chromatography-electrospray ionization tandem MS. R. 2004. Toxicol. Acc. 45 – 54. Guengerich. H. 1493 – 1499. Chem. B. Mahadevan. H. Tyurin. G. F. W. 565 – 569. [335] C. Polhuij. E. Formation and mass spectrometric analysis of DNA and nucleoside adducts by S-(1-acetoxymethyl)glutathione and by glutathione S-transferase-mediated activation of dihalomethanes. G. 157 – 161. Kim. Chirality 1992. H. Toxicol. M. Res. Davis. Lee. Pegram. 19. U. 14. 2001. 1991. 1995. 1999. A. H. R. Chem. 1995. Armstrong. [328] T. J. S. Chem. 97 – 105. V. Dekant. S. Danielson. Evidence for glutathione conjugation as a major pathway of metabolism and implications for isocyanate-mediated toxicities. Mannervik. J. B. V. J. S. J. Marsch. 1997. Baillie. S. S. W. Activation and inactivation of . B. Limited reactivity of formyl chloride with glutathione and relevance to metabolism and toxicity of dichloromethane. B. Biochem. P. Metabolism of dichloromethane to formaldehyde in human erythrocytes: influence of polymorphism of glutathione transferase theta (GST T1-1). Goergens. Ketterer. Chem. H. Evidence for the formation of p-chlorophenyl isocyanate as a reactive intermediate. A. Aust. G. 8. G. 2003. P. G.CHEMISTRY & BIODIVERSITY – Vol. Res. D. A. Wheeler. Han. N. J. 453 – 459. A. D. F. 1995. 1994. Hallier. A. 7. R. Ross. 35. 10. Guengerich. Lee. G. Hashmi. Res. 931 – 936. P. R. Vuilleumier. B. Crease. Chem. W. Bioactivation of [13C]dichloromethane in mouse. Toxicol. M. F. Hartman. [338] S. Glutathione: A vehicle for the transport of chemically reactive metabolites in vivo. Isothiocyanates as substrates for human glutathione transferases: structure-activity studies. Jeon. Lemus. Toxicol. B. Chem. Rose. [331] C. P. Karol. Res. V. F. Res. T. J. J. Chem. M. [330] A. Hiratsuka. Martin. 2006. B. Thier.and dihalomethanes. Chem. Xenobiotica 2005. W. Zhang. Marsch. N. H. W. M. G. T. Hu. McCormick. K. J. 216 – 226. M. Jochheim. 16. [334] F. A. [329] R. 306. P. H. Davis. Anders. A. Res. Day. H. Martin. Forward and reverse catalysis and product sequestration by human glutathione S-transferases in the reaction of GSH with dietary aralkyl isothiocyanates. Res. H. Guengerich.

E. Heinrikson. J.-Biol.1-dichloroethylene to a reactive epoxide in human lung and liver . Comparative toxicity of (þ)-(R). Characterization of glutathione conjugates of the remoxipride hydroquinone metabolite NCQ-344 formed in vitro and detection following oxidation by human neutrophils. N. Commandeur. de Groot. van Bladeren. 12. J. Dybing. J. 446 – 459. H. Chem. 1997. A. A. D. T. 257 – 272. N-Acetyl-S-(1. and tissue-dependent differences. B. 3 – 21. Xenobiotica 1992. Res. A. 155 – 164. Chirality 1995. R. J. D. Res. Ther. 325 – 334. K. 4-Substituted 1-chloro-2-nitrobenzens: SAR and extension of the substrate model of rat glutathione S-transferase 4-4. Connors. Kouzi. E. J. E. 17. Elfarra. J. He.2. Chen. E. Metabolism of 1. 57. B. Conjugation of polychlorinated agrochemical sulfoxides and sulfones by glutathione. Baillie. Boone. van Ommen. Wobbes. Parker. E. A. P. E. L.2-dibromo-3-chloropropane. H. J. J. Fujioka. Glutathione S-transferase conjugation of organophosphorus pesticides yields S-phospho-. A. Toxicol. M. Chem. 359 – 364. and potential applications of glutathione Stransferase-catalyzed cleavage of sulfonamides. Relevance of the mercapturic acid pathway. J. 29. 1999. Toxicol. S. Pharmacol. Klasson-Wehler. A. E. von Euler-Chelpin. N. T. F. S. J. Drug Metab. G. A. C. J. Theoretical study of the SNV reaction of trichloroethylene (TCE) and CH3S À as a model for glutathione conjugation of TCE. S. Chem. Z. D. L. 333 – 340. Dulik. Toxicol. M. A. Comparative disposition and metabolism of 1. Metabolism and kinetics of trichloroethylene in relation to toxicity and carcinogenicity. H. C. Søderlund. 22. A. Pharm. Commandeur. S. J. E. Meerman. B. Xie. 10. T. Res. 28. J. S. Huwe. van Ommen. Ploemen. 411 – 418. and S-alkylglutathione derivatives. Conradi. P. J. Toxicol. Metabolism of pantoprazole involving conjugation with glutathione in rats. 531 – 543. Parker. L. Hueni. Shipley. Burton. B. E. D. Krause. B. Suarato. T. Drug Metab. J. 1999. G. T. Toxicol. Zao. 5 (2008) [339] [340] [341] [342] [343] [344] [345] [346] [347] [348] [349] [350] carcinogenic dihaloalkanes and other compounds by glutathione S-transferase 5-5 in Salmonella typhimurium tester strain NM5004. S-aryl-. E. 20. A. 10. Dybing. C. Ketterer. Meyer. 8. A. S. 638 – 646. Mechanism. J.-G. 41 – 48. 1996. 697 – 713. Chem. S.-Y. Ulreich.2-trichlorovinyl)-l-cysteine and 2. 4971 – 4981. T. K. D. 1997. W. Interact. 27. J. T.2dibromo-3-chloropropane by glutathione S-transferases. Overstreet. Vervoort. 35. M. E. M. 2000. Lindsay. Svensson. P. J. Goeptar. Soffers. D. M.2334 CHEMISTRY & BIODIVERSITY – Vol. X. P. Richard. D. F. Teffera. L. M. L. Res. Reid. A.2. Cytochrome P450dependent bioactivation of 1. species-.and (À)-(S)-1. G. Chem. Koeplinger. Res. 2005. M. A. Søderlund. Toxicol. Exp. A. Xenobiotica 2005. M. Y. Cummings. Z. Holme. Beconi. Dispos. Dispos. Role of cytochrome P450 and glutathione S-transferase a in the metabolism and cytotoxicity of trichloroethylene in rat kidney. Lash. I. Erve. P. Biochemistry 1990. F. C. Chem. Dekant. M.2trichloroethanol: two novel metabolites of tetrachloroethene in humans after occupational exposure. R. Casida. Metabolism and disposition of moxonidine in Fischer 344 rats. Dybing. J. Zhong. Qian. 2007. M. E. Jin. Lash. Res. N. Tomasselli. Dispos. Toxicol. L. Biochem. Nelson. Edmonson. Toxicol. H. Peterson. Abraham. Chem. G. M. 2004. Shang. Chem. 341 – 349. E. 318 – 327. J. 1996. Res. M. Bouwman. Dispos. A. Bakke. A. Chay.3-trichloropropane in rats and mice. 1991. Søderlund. Drug Metab. N. T. P. T. A. G. Drug Metab. Mechanism of the chemoprotective agent diallyl sulfide to glutathione conjugates in rats. H. Regioselectivity and QSAR for the conjugation of a series of fluoronitrobenzenes by purified glutathione S-transferase enzymes from rat and man. 7. Fisher. 1211 – 1217. Moonen. K. Nelson. 1995. N. L. C. 1995. 297. J. A. Vermeulen. L. van der Aar. 1996. Dowsley. S. 439 – 449. R. Metabolic activation of a pentafluorophenylethylamine derivative: formation of glutathione conjugates in vitro in the rat. S. van Bladeren. Chem. J. M. Stearns.2-dibromo-3-chloropropane: evidence for the formation of reactive episulfonium ion intermediates. Vermeulen. 564 – 571. 2001. J. 19. Pharmacol. N. L. Fenselau. Nelson. 9. Czeskis. S. Metabolic activation of 1. J. Xu. P. A. Pharmacol. Birner. 2000. 59. D. M. S. J. Forkert. Putt. K. A. G. Doss. 9.2. 24. M. M. H. 97. 308 – 316. Petsikas. Pearson. J. Shim. P. R. E. Rutkowska. W. J. structure-activity studies. R. D. H. Burka. B. Res. Renal and hepatic toxicity of trichloroethylene and its glutathione-derived metabolites in rats and mice: sex-. Rietjens. Bijloo. R. J. Mahmood. 992 – 998.

-K. Redox in redux: emergent roles for glutathione S-transferase P (GSTP) in regulation of cell signaling and S-glutathionylation.-K. A.CHEMISTRY & BIODIVERSITY – Vol. J. Sonnenbichler. D. N. S. 669 – 677. Dabrowiak. van Ommen. Wang. Arnold. Chem. Chem. A. 709 – 712. Tang. Biol. M. Chem. Interaction of g-glutamyltransferase with clofibryl-S-acyl-glutathione in vitro and in vivo in rats. A. D. Chem. Gregus. Glutathione transferase omega 1 catalyzes the reduction of S(phenacyl)glutathiones to acetophenones. 308. 2004. Pharmacol. M. Gallemann. J. 290. Otagiri. 1992. Chem. K. Lu. 82. F. Methylmercury-cysteinylglycine constitutes the main form of methylmercury in rat bile. J. Reactions of nitrosonitrobenzenes with biological thiols: identification and reactivity of glutathion-S-yl conjugates. Ishikawa. 27. Interact. Chem. Adv. N. W. Naganuma. Hatledid. P. E. Letkeman. 1996. H. Drug Metab. K. Souid. Ballatori. Souid. W. C. Toxicol. 71. S. Barber. Chem. A. J. Toxicol. 641 – 648. Ther. 2002.-F. C. 1257 – 1269. Pharm. 2000. 49 – 67. P. 1988. Hambley. J. M. 3052 – 3059. Pathol. Schäfer. 59. Kinetic studies on the reactions of platinum drugs with glutathione. D. H. M. The inhibition of glutathione S-transferases: mechanisms. Cyurasics. Exp. Several glutathione S-transferase isozymes that protect against oxidative injury are expressed in human liver mitochondria. 1999. Pearson. Pharmacol. S. P. M. 2003. Kinetic study on the reaction of cisplatin with thiols. Z. 9. J. Studies on the metabolism of troglitazone to reactive intermediates in vitro and in vivo. 2007. T. Platinum(IV) antitumour compounds: their bioinorganic chemistry. M. Res. 1999. E. Elsmore. Electrophilic intermediates in the reaction of glutathione and nitrosoarenes. M. Pharmacol. Ther. K. G.-Biol. J. 2007. 1994. A. Coord. 102 – 106. G. 35 – 46. J. K. Kazanis. Townsend.1-dichloroethylene epoxide in murine liver: identification of its glutathione conjugate 2-S-glutathionyl acetate. A. J. R. Imura. D. Kinetic analysis of the covalent binding of captopril to human serum albumin. Glutathione mercaptides as transport forms of metals. Sugii. Pharmacol. 1993. 2001. Oram. Q. Res. J. Sonnenbichler. Sci. Benet. Additional pathways of S- . Correia. Kassahun. Toxicol. 2006. P. Goodisman. Forkert. MacKenzie. D. T. 27 – 37. Toxicol. Sugiyama. 1619 – 1628. N. Doss. Eyer. Binding of dimethylarsinous acid to Cys-13a of rat hemoglobin is responsible for the retention of arsenic in rat blood. H. Hill. Carter. X. Res. J. 1991. D. I. Drug Metab. 197 – 210. 268. Dispos. 14. 1378 – 1384. Harada. P.  HPLC and MS characterization of the nephrotoxic biotransformation products of cisplatin. Glutathione-associated cis-diamminedichloroplatinum(II) metabolism and ATP-dependent efflux from leukemia cells. Marto. R. Baillie. Dispos. J. 2002. Exp. C. M. 86. M. C. 31. 1993. Soc. X. 232. A. P. P.-U. McIntosh. 705 – 713. R. L. Res. J. 73. 6. 151 – 163. Am. Commun. Ali-Osman. Chem. Goodisman. W. P. Grillo. 1997. 4. Chem. toxic consequences and therapeutic benefits. Cashman. Tew. 1991. A. Toxicol. Fernando. K. 62 – 70. 30. In vivo formation and location of 1. Evidence for novel biotransformation pathways involving quinone methide formation and thiazolidinedione ring scission. E. D. Fang. D. Wagner. 14. M. M. Reactions of arsenic(III) and arsenic(V) species with glutathione. S. 271 – 298. Foster. Anders. Res. Toxicol. B. Hanigan. J. 5 (2008) 2335 [351] [352] [353] [354] [355] [356] [357] [358] [359] [360] [361] [362] [363] [364] [365] [366] [367] microsomes. Scott. P. Urano. Maltby. MacDonald. Biochem. Cohen. Decker. Hagrman. I. L. N. M. Board. 114. R. J. X. Pharmacol. Z. 20116 – 20125. Greif. 1033 – 1040. Le. Narazaki. Role of glutathione in the biliary excretion of the arsenical drugs trimelarsan and melarsoprol. Biochem. 289. Chem. 1299 – 1306. Letkeman. D. 20. Formation of glutathionylspironolactone disulfide by rat liver cytochrome P450 or hog liver flavin-containing monooxygenases: a functional probe of two-electron oxidations of the thiosteroid?. 51. D. Toxicol. T. Wang. Pharmacol. D. T. The formation constants of mercury(II)-glutathione complexes. Pharmacol. K. 60. 20. P. 1375 – 1385. 2007. L. Rev. Pharmacol. Ther. McClelland. Deng. Gallagher. van Bladeren. Res. K. Res. I. 1992. Res. Li. Chem. 149 – 154. 215 – 219. Hall. Ther. M.-G. Exp. A. Biochem. T. A. 658 – 666. G. 2001. J. Gardner. W. D. Leung. Buhrow. Dean. Chem. L. Pharmacol. Ellis.

1995. Toxicol. Yamamoto. K. W. J. Mayr. 1998. J. 2006. T. Y. S. Dispos. 123 – 159. Chem. M. Musgrave. Gray. Leeder. Hill. K. Tsuji. Fukushima. M. 17. Ronfeld. R. Reed-Hagen. Drug Metab. Tremaine. C. 13. Senda. Kaufman. 1982. F. A. J. The reaction of amines with carbonyls: its significance in the nonenzymatic metabolism of xenobiotics. 1982. Chiba. J. K. M. Tremaine. Talseth. M. 20. 34. J. C. Ludden. McNay. Krueger. Blackmore. 133 – 134. Nishigaki. A. Zandi. J. Tateishi. P. 222. Ronfeld. R. Miller. Drug Metab. S. J. Meropenem exists in equilibrium with a carbon dioxide adduct in bicarbonate solution. M. Almarsson. E. W. L. C.2336 CHEMISTRY & BIODIVERSITY – Vol. Obach. R. M. 1998. H. L. 1989. in vivo and in vitro. Funke. van Tilborg. W. H. Metabolism and disposition of the 5-hydroxytryptamine uptake blocker sertraline in the rat and dog. Miller. Holmes. M. 1H NMR spectroscopic studies on the reactions of haloalkylamines with bicarbonate ions: formation of N-carbamates and 2-oxazolidones in cell culture media and blood plasma. 159 – 165. L. Ö. 58 – 63. Received June 20. M. C. OConnell. 121 – 130. Dispos. T. S. 8. 1989. H. 31. S. J. 11. Pharmacol. Drug Metab. S. Y. Coe. On the formation of carbamate glucuronides. Haegele. H. Drug Metab. Chem. Delbressine. S. N. 19. 900 – 906. Exp. S. A. a selective a4b2 acetylcholine receptor partial agonist. M. 1991. Drug Metab. Xenobiotica 1990. A. Stong. Drug Metab. Reactions of the nitroso and hydroxylamine metabolites of sulfamethoxazole with reduced glutathione – Implications for idiosyncratic toxicity. Toda. T. S. Petrich. M. M. Mexiletine carbonyloxy b-dglucuronide: a novel metabolite in human urine. Wu. D. Kato. T. J. M. G. D. J. 542 – 550. Xenobiotica 2003. T. P. B. Kadota. 17. 1411 – 1422. Spielberg. Metabolism and disposition of varenicline. Kuroiwa. P. Williams. E. Endogenous generation of hydralazine from labile hydralazine hydrazones. E. McDonnell. A. Dispos. J. Pharm. J. ODonnell. 2008 . Igarashi. M. Stroh. E. Kobayashi. 1046 – 1053. Welch. L. Ther. Kaspersen. A. W. Res. Dispos. A. L. Characterization of a carbamic acid ester glucuronide of the secondary amine sertraline. P. T. Cribb. Fukumoto. G. M. R. B. K. 663 – 666. Sci. Hayakawa. Nicholson. S. 2003. 87. Anthony. H. Obach. J. 871 – 884. 5 (2008) [368] [369] [370] [371] [372] [373] [374] [375] [376] [377] conjugate formation during interaction of 4-nitrosophenethole with glutathione. 33. Metabolism and disposition of novel des-fluoro quinolone garenoxacin in experimental animals and an interspecies scaling of pharmacokinetic parameters. K. Rev. Res. J. Toxicol. Dispos. J. 1409 – 1418. Clementi.