Types of Compounds Forming Glucuronides

TYPE EXAMPLES CH3

O
H N
N CH3

O-Glucuronide
OH HO O OH
Phenols Acetaminophen morphine
OH H
OH Cl O N CH3
H
N
Cl CH3
O
O2N OH

Alcohols Chloramphenicol Propranolol

OH

O O

Enols Hydroxycoumarine
CH3
N
H2N S NHOH
O2 OH
N-hydroxyamines/amides
N-hydroxydapsone N-Hydroxy-2-acetylaminoflourene
 COOH

OH Salicylic acid
Aryl acids
CH3
O

OH

O
Fenoprofen
Arylalkyl acids

N
O
N-Glucuronides NH2 O O CH3
H S
N N
H CH3
N 7-Amino-5- Sulfonamides
Arylamines H2N
O2N nitroindazole Sulfisoxazole

H
N N
Alkylamines CH3
N
CH3 3o Amines
Desipramine
O
NH2
H3C O
Cyproheptadine
Amides Meprobamate
O
H3C NH2
O
 N
S-Glucuronides HS
N
CH3 Methimazole
Sulfhydryl
H3C
S
H3C N
Carbodithioic acid
SH
Disulfirum (reduced form)

C-Glucuronides N
CH3
N

O Phenylbutazone
 Sulfate Conjugation
!  Occurs less frequently than does glucuronidation presumably due to
fewer number of inorganic sulfates in mammals and fewer number of
functional groups (phenols, alcohols, arylamines and N-hydroxy
compounds)
!  Three enzyme-catalyzed reactions are involved in sulfate conjugation

O O O O
- -
O S O P O ATP ADP O S O P O RXH PAP
O ATP PPi Ad - Ad O
- O O
O O O O
- -
O S O- +2 Mg
+2 Sulfotransferase O S XR
Mg (soluble)
O APS phosphokinase O
ATP sulfurylase -2
Sulfate HO OH O3 PO OH Sulfate
Adenosine-5'- 3'-phosphoadenosine-5'- conjugate
phosphosulfate (APS) phosphosulfate (PAPS)
 Sulfation of Drugs

!  Phenolic sulfation predominates

!  Phenolic O-glucuonidation competes favorably with sulfation due to limited
sulfate availability
!  Sulfate conjugates can be hydrolyzed back to the parent compound by
various sulfatases
!  Sulfoconjugation plays an important role in the hepatotoxicity and
carcinogenecity of N-hydroxyarylamides
!  In infants and young children where glucuronyltransferase activity is not
well developed, have predominating O-sulfate conjugation
!  Examples include: α-methyldopa, albuterol, terbutaline, acetaminophen,
phenacetin

H OH H OH H
HO N N CH3 HO N CH3
H HO CH3
CH3
H3C COOH
CH3 CH3
HO HO
OH
α-Methyldopa Albuterol Terbutaline
Possible Mechanism of Phenacetin Toxicity

Electrophilic nitreneum
 Amino Acid Conjugation

!  The first mammalian drug metabolite isolated, hippuric acid, was the
product of glycine conjugation of benzoic acid
R O R O O
COH CONHCH2COH

Benzoic Acid, R = H Hippuric Acid, R = H

Salicylic Acid, R = OH Salicyluric Acid, R = OH

!  Amino acid conjugation of a variety of caroxylic acids, such as aromatic,

arylacetic, and heterocyclic carboxylic acids leads to amide bond formation
!  Glycine conjugates are the most common
!  Taurine, arginine, asparagine, histidine, lysine, glutamate, aspartate,
alanine, and serine conjugates have also been found
 Mechanism of Amino Acid conjugation

Drug-COOH An Acyl-CoA Intermediate

Glycine Conjugate R = H
Glutamine Conjugate R = CH2CH2CONH2
 Brompheniramine Metabolism

CH3 CH3
N NH NH2
N CH3 N N N CHO N COOH
P450 P450 P450 Aldehyde
dehydrogenase

Br Br Br Br Br
Brompheniramine Carboxylic Acid metabolite

Glycine
CH3 N-acyltransferase
H
N N COOH
N CH3 N
O

Br Br
Brompheniramine N-oxide Glycine conjugate
 Glutathione Conjugation
NH 2 O
H
HO N OH
N
HS H
O O O O O O
H S
N S
HO N OH O O O
H H
O NH 2 N
HO N OH
H
NH 2 O
Glutathione reduced form (GSH) Glutathione oxidized form (GSSG)

!  Glutathione is a tripeptide (Glu-Cys-Gly) – found virtually in all

mammalian tissues
!  Its thiol functions as scavenger of harmful electrophilic parent drugs
or their metabolites
!  Examples include SN2 reaction, SNAr reaction, and Michael addition
 SN2 Examples

GSH -
A. R X Y R X SG + Y SN2 X = C, O, S; Y = leaving group or epoxide
Glutathione-
S-Transferase

1. CH3O2SO -
SG CH3O2SO SG
OSO2CH3 S+ G
Busulfan
ONO2 ONO2 ONO2
2.
ONO2 - ONO2 - ONO2 + GSSG
H SG H SG H
O NO2 O SG OH

Nitroglycerine
CH 3
O
O
O N
O O O
CH3
Naproxcinod
 SNAr Examples

X SG
GSH
B. SNRr
Z Z
O -
+ -
O
N N O N +
N O-
1. N SG
S N N NO2 SH
H3C -
SG H3C S
N N N
N N N +
N SG
H3C N
N N N
N N 1-Methyl-4-nitro-5- H
H H
Azathioprine (S-glutathionyl) 6-Mercaptopurine
imidazole
 Michael Addition
H+
-
C. Z SG SG Michael Addition
Z

CH3 CH3 CH3

N N N
-
SG SG

HO O OH HO O O HO O OH

CH3 CH3
-
SG N GS N

O O
O OH HO OH
 Mercapturic Acid Conjugates

Drug Drug
Amino Acid
S (AA) γ-Glutamyl-AA S
O O O O
H H
N N
HO N OH HO NH 2
H γ -Glutamyl
O NH 2 transpeptidase O
Glutathione Conjugate

Drug Acetyl
Glycine S Drug
CoA CoASH
S
O
HO
Cysteinyl NH 2 H 2N
Glycinase N CH 3
O H
S-substituted O
Cysteine Mercapturic
Derivative acid conjugate
 Acetyl Conjugation
!  Metabolism for drugs containing a primary amino group, (aliphatic and aromatic
amines), amino acids, sulfonamides, hydrazines, and hydrazides
! The function of acetylation is to deactivate the drug, although N-
acetylprocainamide is as potent as the parent antiarrhythmic drug procainamide
(Procanbid) or more toxic than the parent drug, e.g., N-acetylisoniazid
! Acetylation is two-step, covalent catalytic process involving N-acetyl transferase

O O O
CoASH
H2N R
H3 C SCoA H3 C X H3 C NHR
X-
X-
N-Acetylation of amines
Genetic polymorphism in N-acetyltransferase activity
Multiple NAT2 alleles (NAT2*5, *6, *7, and *14) have substantially decreased
acetylation activity and are common in Caucasians and populations of African
descent. In these groups, most individuals carry at least one copy of a slow
acetylator allele, and less than 10% are homozygous for the wild type (fast
acetylator) trait. The ratio of NAT2 activity is 7 in Caucasians to 18 in the Chinese
population.
Example of Acetylated Drugs

O O

HO S OH
NH O NH2

CH3
Cilastatin
CH3

HO
H
N NH
S
H3C N
O
COOH Imipenem
 Fatty Acid and Cholesterol Conjugation

!  Hydroxyl-containing drugs can undergo conjugation with a wide

range of endogenous fatty acids such as saturated acids from
C10 to C16 and unsaturated acids such as oleic and linoleic acids
!  Cholesterol ester metabolites have been detected for drugs
containing either an ester or a carboxylic acid

O Cl
O N
Cl
HO OH
Cholesterol
Fatty Acid
Cl O (CH2) 10 COOH

O Prednimustine
 Methyl Conjugation
!  Minor conjugation pathway, important in biosynthesis of epinephrine
and melatonin; in the catabolism of norepinephrine, dopamine,
serotonin, and histamine; and in modulating the activities of
macromolecules (proteins and nucleic acids)
!  Except for the formation of quarternary ammonium salts, methylation
of an amine reduces the polarity and hydrophilicity of the substrates
!  A variety of methyl transferase, such as COMT (catechol O-methyl
transferase), phenol-O-methyltransferase, N-methyl transferase, S-
methyltransferase etc are responsible for catalyzing the transfer of
methyl group from SAM to RXH
H2 N H2N COOH H2 N COOH
COOH
ATP PPi + Pi
Methyltransferase
CH3 -X-R +
H3CS Methionine S + HX-R S
adenosyltransferase CH3 Ad
Ad O
O
Mthetionine

HO OH
HO OH
S-Adenosylmethionine
Mechanism of methyl conjugation
 Case Study
Case 2. Imagine yourself as a drug information specialist at a poison control center.
A technician from the coroner’s office is investigating a case and requires assistance
in identifying the possible sources of benzodiazepines (BZDs) in the toxicology
profile of a particular corpse. The technician has identified four distinct BZDs in this
blood sample. She believes that the major component is diazepam (1) (72% of the
identified BZDs) and that the remaining three components are metabolites (NOTE:
the assay identifies only active compounds). H
N
O

Cl N

Q. What are the three structures of CH3

O H O
potential ACTIVE metabolites for N N
2 OH
diazepam? Cl N Cl N

CH3
O
1 N
4
OH
Cl N

3
 Study Guide

1.  What Roles are Played by Drug Metabolism? Know with structural
examples
2.  Role of stereochemistry in metabolism of drugs with example of warfarin,
ibuprofen and itomidate
3.  What is first pass effect; enterohepatic circulation? Why and how they
occur? Drug examples
4.  Metabolisms in the intestinal mucosa
5.  CYP450, Hepatic microsomal flavin containing monooxygenases (MFMO
or FMO) Monoamine Oxidase (MAO) and Hydrolases. Drugs metabolised
by these enzymes and the active sites of these enzymes. Types of
metabolic reaction catalyzed by these enzymes
6.  Specific CYP enzymes with the number of drugs they metabolize
7.  Few CYP family with their main functions
8.  Drug interaction basics related to metabolic enzymes
 Study Guide Cont.
9.  Mechanism and routes of aromatic hydroxylation. The effects of electron
donating and withdrawing groups in aromatic hydroxylation. Drug examples.
What is NIH shift?
10. Oxidation of olefins. Role of epoxide hydrolase. Can olefenic epoxide be
converted to alcohol as in aromatic epoxide by NIH shift?
11. What type of C in a drug molecule can not be hydroxylated?
12. What is allylic and benzylic hydroxylation? Show drug examples.
13. Show the drug examples where hydroxylation occur on Cα to C=O and C=N
bonds
14. Show the drug examples where hydroxylation occur at aliphatic and alicyclic
carbon atoms. Which carbons are more easily hydroxylated?
15. What is N-oxidatin and N-dealkylation. What enzymes are involved? How do
you differentiate between N-dealkylation and deamination. Drug examples.
What types of drugs generates lactams instead of causing dealkylation?
16. What is the difference between mixed function oxidases and amine oxidases?
 Study Guide Cont.

17. What is the difference between ethanol oxidation and O-dealkylation?

18. What is S-dealkylation, desulfuration and S-oxidation? Drug examples.
19. How does steric factors influence S- O- and N-dealkylations?
20. Oxidative dehalogenation with special example of chloramphenicol. Why
chloramphenicol cause toxicity to the babies?
21. What is MFMO and its active site? What types of functional groups are
metabolized by this enzyme? Drug examples.
22. MAO, dehydrogenases, xanthene oxidases and their functions with drug
examples. Difference between MAO-A and MAO-B.
23. Alcohol and aldehyde dehydrogenases, the coenzymes and the types of drugs
they work on.
24. Azo and nitro reductases, their coenzymes and the drugs they act on.
 Study Guide Cont.
25. Different types of hydrolytic enzymes. Compare rate of hydrolysis of esters,
amides, carbonates and carbamates.
26. What are prodrugs and antedrugs? What are the advantages? Examples.
27. What are different types of conjugation reactions?
28. The enzymes and substrates involved in glucuronidation, and sulfate
conjugation.
29. Why acetaminophen is toxic to neonates? Mechanism of phenacetin and
acetaminophen toxicity.
30. What types of drugs or metabolites may form glycin conjugates?
31. What are different mechanisms involved in glutathione conjugation? What is
mercapturic acid conjugate? Mercapturic acid conjugate of acetaminophen is a
sign of its toxicity – why?
32. Mechanism of acetylation. What is slow and fast acetylator?
33. What is COMT? What coenzymes is involved in its action? What types of drugs
and/or neurotransmitters are metabolized by COMT?
 Acknowledgements

Faculty.swosu.edu/faruk.../PHARM4515-5%20(Drug%20Metabolism)!

Books: 1. Wilson and Gisvold
s Textbook of Organic Medicinal and

Pharmaceutical Chemistry 11th ed. Lippincott, Williams & Wilkins ed.
2. Foye
s Principles of Medicinal Chemistry