The Organic Chemistry of Drug Design and Drug Action

Chapter 8 Prodrugs and Drug Delivery Systems

Prodrugs and Drug Delivery Systems
Prodrug - a pharmacologically inactive compound that is converted to an active drug by a metabolic biotransformation Ideally, conversion occurs as soon as the desired goal for designing the prodrug is achieved. Prodrugs and soft drugs are opposite: • a prodrug is inactive - requires metabolism to give active form • a soft drug is active - uses metabolism to promote excretion A pro-soft drug would require metabolism to convert it to a soft drug

Utility of Prodrugs
1. Aqueous Solubility - to increase water solubility so it can be injected in a small volume 2. Absorption and Distribution - to increase lipid solubility to penetrate membranes for better absorption 3. Site Specificity - to target a particular organ or tissue if a high concentration of certain enzymes is at a particular site or append something that directs the drug to a particular site---often tried to limit the toxicity of anticancer drugs.

Utility of Prodrugs (cont’d)
4. Instability - to prevent rapid metabolism; avoid first-pass effect 5. Prolonged Release - to attain a slow, steady release of the drug 6. Toxicity - to make less toxic until it reaches the site of action 7. Poor Patient Acceptability - to remove an unpleasant taste or odor; gastric irritation 8. Formulation Problems - to convert a drug that is a gas or volatile liquid into a solid

Types of Prodrugs
Drug Latentiation - rational prodrug design
I. Carrier-linked prodrug A compound that contains an active drug linked to a carrier group that is removed enzymatically A. bipartate - comprised of one carrier attached to drug B. tripartate - carrier connected to a linker that is connected to drug C. mutual - two, usually synergistic, drugs attached to each other II. Bioprecursor prodrug A compound metabolized by molecular modification into a new compound, which is a drug or is metabolized further to a drug - not just simple cleavage of a group from the prodrug—e.g., amine getting oxidized to CO2H, to afford the active.

Protecting Group Analogy for the Concept of Prodrugs
Scheme 8.1
A. RCO2H EtOH HCl Δ reaction on R RCO2 Et reaction on R R'CO2Et H 3O + Δ R'CO2H

analogous to carrier-linked
The ester group can be used To modify properties of The drug

B.

RCH=CH2

R'CH=CH 2

1. O3 2. H2O 2

R'CO2H

analogous to bioprecursor

Keep in mind that a prodrug whose design is based on rat metabolism may not be effective in humans.

Mechanisms of Prodrug Activation
Carrier-Linked Prodrugs
Most common activation reaction is hydrolysis. Rate of hydrolysis can be modified by locating alkyl groups in area of the carbonyl group to Increase steric hindrance, and retard hydrolysis rate.

Ideal Drug Carriers
1. Protect the drug until it reaches the site of action 2. Localize the drug at the site of action 3. Allow for release of drug 4. Minimize host toxicity 5. Are biodegradable, inert, and nonimmunogenic 6. Are easily prepared and inexpensive 7. Are stable in the dosage form

Carrier Linkages for Various Functional Groups
Alcohols, Carboxylic Acids, and Related Groups Most common prodrug form is an ester • esterases are ubiquitous
• can prepare esters with any degree of hydrophilicity or lipophilicity • ester stability can be controlled by appropriate electronic and steric manipulations

Prodrugs for AlcoholContaining Drugs
Table 8.1
O

Drug—OH

Drug—OX
Effect on Water Solubility (R = aliphatic or aromatic) decreases (increases lipophilicity) increases (pKa ~ 8)

alcohols
X

C—R O + C—CH 2NHMe2 O C—CH 2CH2COOO C NH

Ester analogs as prodrugs can affect lipophilicity or hydrophilicity

increases (pKa ~ 5)

increases (pKa ~ 4)

+

PO3= (phosphate ester)

increases (pKa ~ 2 and ~ 6)

O CCH 2SO3-

increases (pKa ~ 1)

To accelerate hydrolysis rate: • attach an electron-withdrawing group if a base hydrolysis mechanism is important • attach an electron-donating group if an acid hydolysis mechanism is important To slow down hydrolysis rate: • make sterically-hindered esters • make long-chain fatty acid esters

Another Approach to Accelerate Hydrolysis
Intramolecular hydrolysis of succinate esters
Scheme 8.2
O
-O

H Drug—O-

OH O

O Drug—OH + O
-OOC

COO-

Drug—O O

Also, acetals or ketals can be made for rapid hydrolysis in the acidic medium of the GI tract.

Enolic hydroxyl groups can be esterified as well.
HOOC
S F O Cl N O H2N oxindole 8.1 OH

O O F O Cl N O H2N 8.2 S

antirheumatic agent

Carboxylic Acid-Containing Groups
Esterify as with alcohols

Maintaining Water Solubility of Carboxylate Prodrugs
O + Drug—C—O—CH 2CH2—NRR'2 8.3

Can vary pKa by appropriate choice of R and R′

Prodrugs for Phosphate- or Phosphonate-Containing Drugs
O P O 2 8.4 O O

Amine Prodrugs
Table 8.2
Drug—NH 2 X Drug—NHX

O CR

+ CCHNH3 R

O

O

O CHAr NAr

C OPh —CH 2NHCAr

Amides are commonly not used because of stability Activated amides (low basicity amines or amino acids) are effective pKa of amines can be lowered by 3 units by conversion to N-Mannich bases (X = CH2NHCOAr)

N-Mannich base (R = CH2NHCOPh) has a log D7.4 two units greater than the parent compound.
OH CH3 NHR phenylpropanolamine hydrochloride (R = H . HCl) 8.5

Another approach to lower pKa of amines and make more lipophilic. Imine (Schiff base) prodrug
OH F N O NH2

Cl progabide 8.6

hydrolyze imine and amide to GABA inside brain

anticonvulsant

A Reductive Carrier-Linked Prodrug Approach
Scheme 8.3
OH Y Drug X 8.7 Z NO2 bioreduction Drug X Y :NH Z Drug XH + Y Z 8.8 OH N

Prodrugs of Sulfonamides
A water soluble prodrug of the anti-inflammatory drug valdecoxib (8.9) has been made (8.10).
Ph N O H 2N O CH3 S O valdecoxib 8.9
Na+ N O S O O parecoxib sodium 8.10 Ph N O CH3

Prodrug Analogs of Carbonyl Compounds
Table 8.3
Drug C=O R C X Y C=NR' C=NOH OR' C OR' O C N H S C N H Drug C R Y X

imines oximes ketals

Examples of Carrier-Linked Bipartate Prodrugs

Prodrug for Increased Water Solubility
HO Me O R Me OH O

O R' = CCH2CH2CO2 Na
OR'

Prodrug forms
R' = PO3Na 2 for aqueous injection or opthalmic use

prednisolone (R = R' = H) methylprednisolone (R = CH3 , R' = H) 8.11

poor water solubility

corticosteroid

Choice of water solubilizing group: The ester must be stable enough in water for a shelf life of > 2 years (13 year half-life), but must be hydrolyzed in vivo with a half-life < 10 minutes. Therefore, in vivo/in vitro lability ratio about 106.

To avoid formulation of etoposide with detergent, PEG, and EtOH (used to increase water solubility), it has been converted to the phosphate prodrug.
CH3 O HO O O O O O HO O O

CH3O

OR

OCH3

etoposide (R = H) etoposide phosphate (R = PO3 H2 ) 8.12

Prodrug for Improved Absorption Through Skin
HO CH3 F O F fluocinolone acetonide (R = H) fluocinonide (R = COCH3 ) 8.14 O CH3 OR O O CH 3 CH3

corticosteroids - inflammation, allergic, pruritic skin conditions

Better absorption into cornea for the treatment of glaucoma
RO RO OH NHCH3

dipivefrin (R = Me3 CCO) epinephrine (R = H) 8.15

The cornea has significant esterase activity

Prodrug for Site Specificity
RO

Bowel sterilant
OR

N H

O

oxyphenisatin (R = H) 8.16

(administer rectally)

prodrug R = Ac (administer orally) hydrolyzed in intestines

Prodrug for Site Specificity
The blood-brain barrier prevents hydrophilic molecules from entering the brain, unless actively transported. The anticonvulsant drug vigabatrin (see Chapter 5) crosses poorly. A glyceryl lipid (8.17, R = linolenoyl) containing one GABA ester and one vigabatrin ester was 300 times more potent in vivo than vigabatrin.
OCOR O O O 8.17 O NH2 NH2

Site Specificity Using Enzymes at the Site of Action

OR

RO = diethylstilbestrol diphosphate (R = PO 3 ) diethylstilbestrol (R = H) 8.18

Phosphatase should release the drug selectively in tumor cells. This approach has not been successful because the prodrugs are too polar, enzyme selectivity is not sufficient, or tumor cell perfusion rate is poor.

Enzyme-Prodrug Therapies
For selective activation of prodrugs in tumor cells Two steps: 1. incorporate a prodrug-activating enzyme into a target tumor cell 2. administer a nontoxic prodrug which is a substrate for the exogenous enzyme incorporated

Criteria for Success with Enzyme-Prodrug Therapies
1. The prodrug-activating enzyme is either nonhuman or a human protein expressed poorly 2. The prodrug-activating enzyme must have high catalytic activity 3. The prodrug must be a good substrate for the incorporated enzyme and not for other endogenous enzymes 4. The prodrug must be able to cross tumor cell membranes 5. The prodrug should have low cytotoxicity and the drug high cytotoxicity 6. The activated drug should be highly diffusable to kill neighboring nonexpressing cells (bystander killing effect) 7. The half-life of the active drug is long enough for bystander killing effect but short enough to avoid leaking out of tumor cells

Antibody-Directed Enzyme Prodrug Therapy (ADEPT)
An approach for site-specific delivery of cancer drugs. Phase One: An antibody-enzyme conjugate is administered which binds to the surface of the tumor cells. The antibody used has been targeted for the particular tumor cell. The enzyme chosen for the conjugate is one that will be used to cleave the carrier group off of the prodrug administered in the next phase. Phase Two: After the antibody-enzyme has accumulated on the tumor cell and the excess conjugate is cleared from the blood and normal tissues, the prodrug is administered. The enzyme conjugated with the antibody at the tumor cell surface catalyzes the conversion of the prodrug to the drug when it reaches the tumor cell.

Advantages: 1. Increased selectivity for targeted cell 2. Each enzyme molecule converts many prodrug molecules 3. The released drug is at the site of action 4. Demonstrated to be effective at the clinical level 5. Concentrates the drug at the site of action Disadvantages: 1. Immunogenicity and rejection of antibody-enzyme conjugate 2. Complexity of the two-phase system and i.v. administration 3. Potential for leakback of the active drug

ADEPT

An example is carboxypeptidase G2 or alkaline phosphatase linked to an antibody to activate a nitrogen mustard prodrug.

Scheme 8.4
I N I carboxypeptidase G2 H N O 8.19 electron-withdrawing; deactivates nitrogen mustard I N I L-Glu + CO 2

+ OH

O

CO2H CO2H

electron-donating; activates nitrogen mustard

Humanization of antibodies minimizes immunogenicity. Note the prodrug-activating enzyme is a bacterial enzyme.

Antibody-Directed Abzyme Prodrug Therapy (ADAPT)
Instead of using a prodrug-activating enzyme, a humanized prodrug-activating catalytic antibody (abzyme) can be used. Ideally, the abzyme catalyzes a reaction not known to occur in humans, so the only site where the prodrug could be activated is at the tumor cell where the abzyme is bound. Antibody 38C2 catalyzes sequential retro-aldol and retro-Michael reactions not catalyzed by any known human enzyme. Found to be long-lived in vivo, to activate prodrugs selectively, and to kill colon and prostate cancer cells.

Abzyme 38C2 Activation of a Doxorubicin Prodrug
Scheme 8.5
O OH O OH OH CH3O O OH H O O CH3 NH O HO 8.20 O BH O 38C2 CH3O O OH H O CH3 HO O O NH O O O -CO2 O OH BH O OH O OH 38C2 OH H O CH3 HO O NH OO O OH O OH OH O

O retro-aldol

CH3O O retro-Michael

+H+ OH

O OH OH

CH3O

O CH3

OH H O O

NH2 HO doxorubicin

Gene-Directed Enzyme Prodrug Therapy (GDEPT) Also known as suicide gene therapy
A gene encoding the prodrug-activating enzyme is expressed in target cancer cells under the control of tumor-selective promoters or by viral transfection. These cells activate the prodrug as in ADEPT.
Cl N O 2N N NMe O O NH O N H OMe Cl OMe OMe nitroreductase H HO N: N NMe O O Cl N O N H OMe NH N O N H OMe OMe OMe

-CO2 +H+

OMe OMe

Scheme 8.6

NH 2 amino-seco-CBI-TMI 8.21

Prodrug for Stability
protection from first-pass effect
Oral administration has lower bioavailability than i.v. injection. O NHCH(CH3 )2
OR'

R propanolol (R = R' = H) 8.22

antihypertension
O prodrug R' = CCH2 CH2COOH

plasma levels 8 times that with propanolol

Prodrugs for Slow and Prolonged Release
1. To reduce the number and frequency of doses 2. To eliminate night time administration 3. To minimize patient noncompliance 4. To eliminate peaks and valleys of fast release (relieve strain on cells) 5. To reduce toxic levels 6. To reduce GI side effects Long-chain fatty acid esters hydrolyze slowly Intramuscular injection is used also

O F N

OR

Cl haloperidol (R = H) haloperidol decanoate (R = CO(CH2) 8CH 3 ) 8.24

Sedative/tranquilizer/antipsychotic
O prodrug R' = C(CH2) 8CH3

slow release

inject i.m. Antipsychotic activity for about 1 month

Prodrugs to Minimize Toxicity
Many of the prodrugs just discussed also have lowered toxicity. For example, epinephrine (for glaucoma) has ocular and systemic side effects not found in dipivaloylepinephrine.

Prodrug to Increase Patient Acceptance
CH3

The antibacterial drug clindamycin (8.28) is bitter and not well tolerated by children.

N

H

H N O OH

Cl

H

O HO

Clindamycin palmitate clindomycin (R = H) clindomycin phosphate (R = PO 3H2) is not bitter.

SCH3 OR

clindomycin palmitate (R = O(CH 2) 14 CH3 ) 8.28

Either not soluble in saliva or does not bind to the bitter taste receptor or both.

Prodrug to Eliminate Formulation Problems
Formaldehyde is a gas with a pungent odor that is used as a disinfectant. Too toxic for direct use.
N CH2O + NH3 H+ H3O+ N N methenamine 8.30 N

It is a stable solid that decomposes in aqueous acid. The pH of urine in the bladder is about 4.8, so methenamine is used as a urinary tract antiseptic. Has to be enteric coated to prevent hydrolysis in the stomach.

Areas of Improvement for Prodrugs
• site specificity • protection of drug from biodegradation • minimization of side effects

Macromolecular Drug Delivery
To address these shortcomings, macromolecular drug delivery systems have been developed.
A bipartate carrier-linked prodrug in which the drug is attached to a macromolecule, such as a synthetic polymer, protein, lectin, antibody, cell, etc. Absorption/distribution depends on the physicochemical properties of macromolecular carrier, not of the drug. Therefore, attain better targeting. Minimize interactions with other tissues or enzymes. Fewer metabolic problems; increased therapeutic index.

Disadvantages of Macromolecular Delivery Systems
• Macromolecules may not be well absorbed • Alternative means of administration may be needed (injection) • Immunogenicity problems

Macromolecular Drug Carriers
Synthetic polymers
CH 2 CH2 CH x OH CH O O O 8.32 O y

Aspirin linked to poly(vinyl alcohol) has about the same potency as aspirin, but less toxic.

Steric Hindrance by Polymer Carrier
poly(methacrylate)
CH3 CH2 C x C=O O CH3 CH2 C y

C=O O

S=O O

to enhance water solubility

testosterone

8.34

No androgenic effect Polymer backbone may be sterically hindering the release of the testosterone.

A spacer arm was added, and it was as effective as testosterone. CH3 CH3
CH2 C x CH2 C y C=O O
-

C=O O spacer arm + Cl NMe2 O O

H3 C

S=O

to enhance water solubility

O

8.35

Poly(α-Amino Acid) Carriers
poly(L-glutamine)
O NH CH C x O O O C CH

spacer
O N H

O 8.37

norethindrone - contraceptive

Slow release over nine months in rats

General Site-Specific Macromolecular Drug Delivery System
Polymer chain

Solubilizer

spacer

Homing device

either hydrophilic or hydrophobic

Drug 8.38

for site specificity

Site-Specific Delivery of a Nitrogen Mustard
O O NHCH C x NHCH C y O NHCH C z

poly(L-Glu)

O

O ONH

spacer arm
O NH Ig

water-solubilizing
Cl N 8.39 Cl

antibody from rabbit antiserum against mouse lymphoma cells

All 5 mice tested were alive and tumor free after 60 days (all controls died). Also, therapeutic index greatly enhanced (40 fold).

Tumor Cell Selectivity
Drug attached to albumin (R = albumin) Tumor cells take up proteins rapidly. Proteins broken down inside cells, releasing the drug. Shown to inhibit growth of Ectomelia virus in mouse liver, whereas free inhibitor did not.
NH2 N RO O N O HO OH cytosine arabinoside (R = H) 8.41

antitumor

Antibody-Targeted Chemotherapy
Scheme 8.7

calicheamicin, except as disulfide instead of trisulfide
HO O CH3O O CH3 H N O gemtuzumab ozogamicin 8.42 S S RSCH3O O S 8.43 NH H O polysaccharide O NH H O polysaccharide HO

antibody

Lys

H N O

N O

humanized

spacer

Does not release calicheamicin nonenzymatically. Exhibits no immune response.

Tripartate Drugs
(Self-immolative Prodrugs)
A bipartate prodrug may be ineffective because the linkage is too labile or too stable. In a tripartate prodrug, the carrier is not attached to the drug; rather, to the linker. Therefore, more flexibility in the types of functional groups and linkages that can be used, and it moves the cleavage site away from the carrier. The linker-drug bond must cleave spontaneously (i.e., be self-immolative) after the carrier-linker bond is broken.

Tripartate Prodrugs
Scheme 8.8
enzyme

Carrier

Linker

Drug

Carrier

+

Linker

Drug

spontaneous

Linker

+

Drug

Typical Approach
Scheme 8.9
O Drug—X—CH2 —O—CR esterase Drug—X—CH2 —Ofast Drug—X+ CH 2O + RCOOH

Tripartate Prodrugs of Ampicillin
O

Poor oral absorption (40%) Excess antibiotic may destroy important intestinal bacteria used in digestion and for biosynthesis of cofactors. Also, more rapid onset of resistance.

NH NH2 O ampicillin 8.44 N

S COO-

antibacterial

Various esters made were too stable in humans (although they were hydrolyzed in rodents) - thought the thiazolidine ring sterically hindered the esterase.

Tripartate Prodrugs of Ampicillin
Scheme 8.10
O Ph NH2 O N O O R O O R' NH Ph S esterase NH2 O N O 8.46 O R 8.44 H O R EtOH + CO 2 O NH S + .. OH R'COOH when R' = OEt

bacampicillin (R = CH3 , R' = OEt) pivampicillin (R = H, R' = t-Bu) 8.45

98-99% absorbed

Ampicillin is released in < 15 minutes

Reversible Redox Drug Delivery System to the CNS
Scheme 8.11
O Drug—X N CH3 8.47

hydrolysis deactivated
O crosses blood-brain barrier Drug—X N CH3 HOOC + N+ CH3 8.49 Drug — XH enzyme oxidation Drug—X

hydrolysis activated
O

N+ 8.48 CH 3

hydrophilic drug

enzyme hydrolysis

electron-donating, lipophilic carrier

electronwithdrawing; hydrophilic

Passive diffusion of 8.47 into the brain; active transport of 8.49 out of the brain XH of the drug is NH2, OH, or COOH If oxidation occurs before it gets into the brain, it cannot cross the blood-brain barrier.

When the drug is a carboxylic acid, a selfimmolative reaction also can be used.
Scheme 8.12
O Drug C OCH2 8.50 O O C carrier esterase O O Drug—C—O—CH2 —O- + HOC fast —CH2O carrier

Drug—COO-

Example of Redox Drug Delivery
Antibody generation in the brain is not significant. β-Lactams are too hydrophilic to cross the blood-brain barrier effectively.
R O H N O N O O O N Me esterase R O H N O N OO S -CH2O R O H N O N O O O S O N Me 8.49 O S O enzymatic oxidation R O H N O N O O O N Me S O

8.51

Scheme 8.13

High concentrations of β-lactams delivered into brain.

Tripartate Prodrug for Delivery of Antibacterials
Permeases are bacterial transport proteins for uptake of peptides.
Scheme 8.14
HN O O CH3 N H 8.53 H + NH 4 + O COO N COOO F 1. permease 2. peptidase + H3N HN COOCH3 + O .. H2N O + H2N H COO
-

O F N COO-

+ H3N

+

HN O N H

F

Only L,L-dipeptides are active

Mutual Prodrugs
A bipartate or tripartate prodrug in which the carrier is a synergistic drug with the drug to which it is linked.
Antibacterial ampicillin
O Ph NH 2 N H O N O sultamicillin 8.59 S O O O O S O N O

β-lactamase inactivator penicillanic acid sulfone

Hydrolysis gives 1:1:1 ampicillin : penicillanic acid sulfone : formaldehyde

Ideal Mutual Prodrugs
• Well absorbed • Both components are released together and quantitatively after absorption • Maximal effect of the combination of the two drugs occurs at 1:1 ratio • Distribution/elimination of components are similar

Bioprecursor Prodrugs
Carrier-linked prodrugs largely use hydrolytic activation Bioprecursor drugs mostly use oxidative or reductive activation The metabolically-activated alkylating agents discussed in Chapter 6 are actually examples of bioprecursor prodrugs.

Protonation Activation
Discovery of Omeprazole Cimetidine and ranitidine (Chapter 3) reduce gastric acid secretion by antagonizing the H2 histamine receptor. Another way to lower gastric acid secretion is by inhibition of the enzyme H+,K+-ATPase (also called the proton pump), which exchanges protons for potassium ions in parietal stomach cells, thereby increasing stomach acidity.

Lead Discovery
Lead compound found in a random screen.
S N 8.60 NH2

Liver toxicity observed thought to be because of the thioamide group.

Lead Modification
Related analogs made, and 8.61 had good antisecretory N activity. S
N 8.61 N H

Modification gave 8.62 with high activity.
N S 8.62 N N H

The sulfoxide (8.63) was more potent, but it blocked iodine uptake into the thyroid.
O N S N N H timoprazole 8.63

Lead Modification (cont’d)
Modification of 8.63 gave 8.64 having no iodine blockage activity.
CH3 N O S N CO2CH3 CH3

Picoprazole shown to be an inhibitor of H+,K+-ATPase. SAR of analogs indicated electron-donating groups on H3 C the pyridine ring were favorable. Increased inhibition of H+,K+-ATPase. Best analog was omeprazole (8.65).

N H picoprazole 8.64

OCH3 CH3 N

O S

N N H

OCH3

omeprazole 8.65

The pKa of the pyridine ring of omeprazole is about 4, so it is not protonated and able to cross the secretory canaliculus of the parietal cell. The pH inside the cell is below 1, so this initiates the protonation reaction below.
Scheme 8.16
H3 C OCH 3 CH3 N .. S HN N H+ OCH 3 O HN N H H3 C OCH3 CH3 N S O .. HN N H3 C OCH3 CH3 N S -H2 O OH N H3 C OCH 3 CH3 N S N S

OCH3 omeprazole 8.65

OCH 3 8.66

OCH 3

+H+

H3 C

OCH 3 CH3 N N S NH S

Formation of 8.66 leads to covalent attachment. Omeprazole also inhibits isozymes of carbonic anhydrase, another mechanism for lowering gastric acid secretion.

OCH 3

Hydrolytic Activation
Hydrolysis can be a mechanism for bioprecursor prodrug activation, if the product requires additional activation.
Scheme 8.17
OS S+ O N H Me S N K2CO3/RX O Me R S OS+ N HO O Me S N

O Me OH Me HO leinamycin 8.70 O

Me

HO 8.71

O

antitumor agent

prodrugs of leinamycin

HOO Me

O O

Hydrolysis of these analogs gives an intermediate that reacts further to the activated form.
OS+ N Me SS N O O Me OS+ N HO O Me S N S OS+ O N H Me S N DNA cleavage Me O O O O O 8.73 O

S O Me

HO

O Me OH

Me

O O 8.72

O

Me

increased stability over leinamycin

O HO2 C Me HO S N H

Me

S N

this was synthesized and gave 8.74 as well as DNA cleavage

Scheme 8.18

HO Me O O 8.74 O

Elimination Activation
Scheme 8.19
B: H N B O N H O H leflunomide 8.75 CH3 CF 3 N O C N H CH3 8.76 CF 3

HO

rheumatoid arthritis drug

potent inhibitor of dihydroorotate dehydrogenase

Inhibition of dihydroorotate dehydrogenase blocks pyrimidine biosynthesis in human T lymphocytes.

Oxidative Activation
N-Dealkylation Sedative 8.20
CH 3 N Cl O X N N N CH3 P450 CH 3 Cl CH3 N O X CH 3 N N N CH3 H P450 Cl N O X N N NH2 ..

CH 3 N Cl

N N -H2 O N X Cl

CH3 N

N N .. NH X

HO

alprazoalam (X = H) triazolam (X = Cl) 8.77

O-Dealkylation
Analgesic activity of phenacetin is a result of O-dealkylation to acetaminophen.
O HN CH3

OR phenacetin (R = CH2 CH3 ) acetaminophen (R = H) 8.78

Oxidative Deamination
Neoplastic (cancer) cells have a high concentration of phosphoramidases, so hundreds of phosphamide analogs of nitrogen mustards were made for selective activation in these cells.
Scheme 8.21
H N P HO: O Cl P-450 H N P O O Cl Cl B: 8.81 H H NH 2 O P O N O N O N 8.80 Cl Cl

Cl cyclophosphamide 8.79

HPO4=

+

NH3

+ HN 8.84

Cl Cl

spontaneous or phosphoramidase

H2N P O O N 8.82 DNA

O Cl + Cl H 8.83

Cyclophosphamide was very effective, but it required liver homogenates (contains P450) for activation. Therefore oxidation is required, not hydrolysis.

DNA O HN H2N N H N

+ N N DNA

O OH HN H 2N N N N 8.86

H N

OH

8.85

isolated

N-Oxidation
identified
O CH3 NHNHCH 2 CNHCHMe 2 P450 CH3 NHNHCH 2 HO procarbazine 8.87 O CNHCHMe 2 O -H2 O CH3N=N—CH + B—H H B: CNHCHMe 2 8.88

advanced Hodgkin’s disease
CH 3N-NH2 + OHC H 8.89

O CNHCHMe 2 H 2O CH3N-N=CH H

O CNHCHMe 2

CH3N=NH + CH3-N CH3 N DNA HN H 2N

O

+N N

CH3 HN H2N

O

CH3 N

+ N2

N DNA

N 8.90

N

Scheme 8.22

identified

N-Oxidation
Pralidoxime chloride is an antidote for nerve poisons. It reacts with acetylcholinesterase that has been inactivated by organophosphorus toxins.

Cl -

+N CH3

N OH

pralidoxime chloride 8.91

To increase the permeability of pralidoxime into the CNS, the pyridinium ring was reduced (8.92).
N CH3 8.92 N OH

oxidation into brain
N OH

N - + Cl CH3

pralidoxime chloride 8.91

Similar to the reversible redox drug delivery strategy for getting drugs into the brain by attaching them to a dihydronicotinic acid, hydrophobic 8.92 crosses the blood-brain barrier; oxidation to 8.91 prevents efflux from brain.

Mechanism of Acetylcholinesterase
Scheme 8.23
O Me 3N—CH 2CH2—O Trp H CH3 Trp Phe B+ :B H O Me 3N—CH2CH2—OH Trp Trp Phe B: O O CH3 + HB

H2O + Me 3NCH2CH2OH + CH3 COOH

Inactivation of Acetylcholinesterase by Diisopropyl Phosphorofluoridate
Scheme 8.24 affinity labeling agent
O Trp Phe Trp F P H B+ O O H 8.93 :B Trp Phe Trp B O O P O O HB O

irreversible inhibition

Inactivation prevents degradation of the excitatory neurotransmitter acetylcholine. Accumulation of acetylcholine causes muscle cells in airways to contract and secrete mucous, then muscles become paralyzed.

Reactivation of Inactivated Acetylcholinesterase by Pralidoxime
Scheme 8.25
O Trp Phe Trp B P O O HB O pralidoxime N Trp Me Phe Trp N H O O: B O O P O HB

O N Me N O P O O

N Trp Me Phe Trp

O N P O O O BH OH :B

Temporary inhibition of acetylcholinesterase enhances cholinergic action on skeletal muscle.
Scheme 8.26
O Me 3N Trp Phe Trp O H B+ O NMe2 :B H Me 3N Trp Trp Phe OH B: O O NMe2 + HB

neostigmine 8.94

Used for the neuromuscular disease myasthenia gravis

covalent, but reversible slow inhibition
Me 3N OH

H 2O

+

CO 2 + Me2NH

Reversible (noncovalent) inhibitors of acetylcholinesterase, such as donepezil and tacrine are used for Alzheimer’s disease. Enhances neurotransmission involved in memory.
H3CO H3CO O NH2 N HCl donepezil hydrochloride 8.95
.

N.

HCl

tacrine hydrochloride 8.96

S-Oxidation
Poor oral bioavailability of brefeldin A. Converted to Michael addition sulfide prodrug (8.98). S-Oxidation and elimination gives brefeldin A.
Scheme 8.27
H HO HO H brefeldin A 8.97 O O CH3 RSH HO H 8.98 H HO RS H O O CH3 [O] HO H :B H HO RS O H H 8.99 H O O CH3

-RSOH

antitumor, antiviral agent

Aromatic Hydroxylation
Cyclohexenones as prodrugs for catechols
Scheme 8.28

N

P450 HO

N

-H2 O

N

N

O 8.100

O

O

OH P450

oxidation next to sp2 carbonyl

aromatic hydroxylation

N HO OH 8.101

Alkene Epoxidation
O

N O NH2
O

N NH2

carbamazepine 8.102

8.103

active anticonvulsant agent

Transamination
Stimulation of pyruvate dehydrogenase results in a change of myocardial metabolism from fatty acid to glucose utilization. Glucose metabolism requires less O2 consumption. Therefore, utilization of glucose metabolism would be beneficial to patients with ischemic heart disease (arterial blood flow blocked; less O2 available).

Arylglyoxylic acids (8.104) stimulate pyruvate dehydrogenase, but have a short duration of action.
O COOH R 8.104

Oxfenicine (8.105, R = OH) is actively transported and is transaminated (a PLP aminotransferase) in the heart to 8.104 (R = OH).
NH2 COOH R oxfenicine (R = OH) 8.105

Reductive Activation
Azo Reduction
Scheme 8.29
COOH NHSO2 N=N sulfasalazine 8.106 OH

N

ulcerative colitis
H 2N

COOH OH 8.107

Anaerobic cleavage by bacteria in lower bowel
+
N NHSO2 sulfapyridine 8.108 NH2

For inflammatory bowel disease

Causes side effects

To prevent side effect by sulfapyridine a macromolecular delivery system was developed.
n

poly(vinylamine)

Not absorbed or metabolized in small intestine.

NH SO2

spacer
N N 8.109

CO2Na OH

CO2Na NH2 OH

Released by reduction at the disease site.

Sulfapyridine is not released (still attached to polymer). More potent than sulfasalazine.

Azido Reduction
R N HO N O OH HO vidarabine (R = NH 2) 8.110 N N

antiviral drug

Vidarabine is rapidly deaminated by adenosine deaminase. 8.110, R = N3 is not a substrate for adenosine deaminase and also can cross the blood-brain barrier for brain infections.

Sulfoxide Reduction
F CO2H CH3

CH3 S

O sulindac 8.111

anti-arthritis

Sulindac is inactive in vitro; the sulfide is active in vitro and in vivo.

Sulindac is an indane isostere of indomethacin, which was designed as a serotonin analog. The 5-F replaced the 5-OMe group to increase analgesic properties. The p-SOCH3 group replaced p-Cl to increase water solubility.
F CO2H CH3

MeO N O

CO2H CH3 HO N H serotonin

NH2

CH3 S

O sulindac 8.111

Cl indomethacin 8.112

Disulfide Reduction
To increase the lipophilicity of thiamin for absorption into the CNS.
Scheme 8.30
N CH3 N N NH2 O H S S OH .. GSH O CH3 N N N NH2 +B O H H SOH

8.113

nonenzymatic
+ N N NH2 thiamin 8.114 S N: N OH S

N CH3

OH CH3

N

NH2 OH

poorly absorbed into CNS

CH3O N HN NH2

To diminish toxicity of primaquine and target it for cells with the malaria parasite, a macromolecular drug delivery system was designed.
Intracellular thiol much higher than in blood; selective reduction inside the cell
lactose N O N H S S NH3+ 8.116 serum albumin

primaquine 8.115

antimalarial, toxic
CH3O

HN

primaquine

for improved uptake in liver

lactose

Therapeutic index of 8.116 is 12 times higher than 8.115 in mice.

Nitro Reduction
Scheme 8.32
O Cl O N H3 C O O 2N 8.118 O O
-O

HN O O O HO N

F bioreduction Cl O N H3 C O .. HO NH P O HO O

O HN O O N F

Cl .. N H3 C O P OHO 8.120 O HN O O

O F N

P O

8.119 O O F O N O HO N H3 C P OHO O HN O O N F

HN O O O HO 8.121 N

F O H 2O O P O

HN O

P O-

Mechanism-based inactivator of thymidylate synthase

Nucleotide Activation
Scheme 8.33
=

O3PO

SH N N N N H

O HO OH

O

O N
=O 3PO

SH N N O HO OH 8.123 N

O—P—O—P—OO
-

O

-

6-mercaptopurine 8.122

hypoxanthine-guanine phosphoribosyltransferase

Anti-leukemia drug

Inhibits several enzymes in the purine nucleotide biosynthesis pathway.

Phosphorylation Activation O
O HN H2N RO N O
HO 8.125

N N

HN H 2N HO N O

N N

acyclovir (R = H) 8.124

antiviral

2′-deoxyguanosine

Resembles structure of 2′-deoxyguanosine Uninfected cells do not phosphorylate acyclovir (selective toxicity) viral thymidine kinase R = PO3= viral guanylate kinase R = P2O6-3 viral phosphoglycerate kinase R = P3O9-4

Acyclovir triphosphate is a substrate for viral α-DNA polymerase but not for normal α-DNA polymerase Incorporation into viral DNA leads to a dead-end complex (not active). Disrupts viral replication cycle and destroys the virus. Even if the triphosphate of acyclovir were released, it is too polar to be taken up by normal cells. High selective toxicity

Resistance to Acyclovir
Modification of thymidine kinase Change in substrate specificity for thymidine kinase Altered viral α-DNA polymerase

Only 15-20% of acyclovir is absorbed Therefore, prodrugs have been designed to increase oral absorption.
Hydrolyzes here
N H2N HO N O 8.126 NH2 N N

Prodrug for a prodrug

adenosine deaminase

acyclovir

Hydroxylates here
N H 2N HO N O 8.127 N N

xanthine oxidase

acyclovir This prodrug is 18 times more water soluble than acyclovir.

A bipartate carrier-linked prodrug of acyclovir, the L-valyl ester of acyclovir, has 3-5 fold higher oral bioavailability with the same safety profile.
O HN H 2N H 2N O O valaciclovir 8.128 N O N N

An analog of acyclovir whose structure is even closer to that of 2′-deoxyguanosine is ganciclovir.
O HN H 2N HO N O N N

HO ganciclovir 8.129

More potent than acyclovir against human cytomegalovirus.

Two carbon isosteres of ganciclovir are available.
O HN H 2N HO N N N
N H 2N AcO N N N

HO penciclovir 8.130

C in place of O

AcO famciclovir 8.131

C in place of O

Better oral absorption than penciclovir Converted to penciclovir rapidly

Sulfation Activation
Activity of minoxidil requires sulfotransferase-catalyzed sulfation to minoxidil sulfate.
OR H2N N N N NH2

minoxidil (R = ) minoxidil sulfate (R = SO 3- ) 8.132

hair growth

Inhibitors of the sulfotransferase inhibit the activity of minoxidil, but not minoxidil sulfate.

Decarboxylation Activation
An imbalance in the inhibitory neurotransmitter dopamine and the excitatory neurotransmitter acetylcholine produces movement disorders, e.g. Parkinson’s disease. In Parkinson’s there is a loss of dopaminergic neurons and a low dopamine concentration. Dopamine treatment does not work because it cannot cross blood-brain barrier, but there is an active transport system for L-dopa (levodopa, 8.133, R = COOH).

HO HO NH2 H R levodopa (R = COOH) 8.133

After crossing bloodbrain barrier

L-aromatic amino acid decarboxylase (PLP)

dopamine (R = H)

Does not reverse the disease, only slows progression.

Combination Therapy
Monoamine oxidase B (MAO B) degrades dopamine in the brain. Therefore, a MAO B-selective inactivator is used to protect the dopamine - selegiline (L-deprenyl). Peripheral L-aromatic amino acid decarboxylase destroys >95% of the L-dopa in the first pass. Maybe only 1% actually gets into brain.

To protect L-dopa from peripheral (but not CNS) degradation, inhibit peripheral L-aromatic amino acid decarboxylase with a charged molecule that does not cross the blood-brain barrier.
HO HO H3 C NHNH3 COO+

HO HO

OH O N N H H benserazide 8.135 NH2 OH

carbidopa 8.134

(used in U.S.)

(used in Europe and Canada)

Selectively inhibits peripheral L-amino acid decarboxylase. The dose of L-dopa can be reduced by 75%.

Selective Inactivation of Brain Monoamine Oxidase A
Earlier we found that inactivation of brain MAO A has an antidepressant effect, but a cardiovascular side effect occurs from inactivation of peripheral MAO A. 8.136 (R = COOH) is actively transported into the brain, where L-aromatic amino acid decarboxylase converts it to 8.136 (R = H), a selective MAO A mechanism-based inactivator. Carbidopa protects 8.136 (R = COOH) from F decarboxylation outside of the brain.
NH2 R OH 8.136

Selective Delivery of Dopamine to Kidneys
Dopamine increases renal blood flow. Prodrugs were designed for selective renal vasodilation.
Scheme 8.34
COO+ NH 3 O H N COO OH OH L-γ-glutamyl transpeptidase OH + NH 3 COOOH

8.137

Glu

L-dopa
L-aromatic amino acid decarboxylase + NH3 OH OH

L-γ-glutamyl-L-dopa

Dopamine accumulates in the kidneys because of high concentrations of L-γ-glutamyltranspeptidase and L-aromatic amino acid decarboxylase there.

Example of a carrier-linked prodrug of a bioprecursor prodrug for dopamine.