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
EtOH reaction H 3O + analogous to
A. RCO2H RCO2 Et R'CO2Et R'CO2H
HCl on R Δ carrier-linked
Δ The ester group can be used
To modify properties of
The drug

reaction 1. O3 analogous to
B. RCH=CH2 R'CH=CH 2 R'CO2H
on R 2. H2O 2 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
 Drug—OH Drug—OX
Prodrugs for Alcohol- alcohols
Containing Drugs X Effect on Water Solubility

O
Table 8.1 (R = aliphatic or aromatic)
C—R decreases (increases lipophilicity)
Ester analogs as O
+ increases (pKa ~ 8)
prodrugs can affect C—CH 2NHMe2

lipophilicity or O
increases (pKa ~ 5)
C—CH 2CH2COO-
hydrophilicity
O
increases (pKa ~ 4)
C
NH
+
increases (pKa ~ 2 and ~ 6)
PO3= (phosphate ester)

O
increases (pKa ~ 1)
CCH 2SO3-
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
H OH O
-O
- -OOC COO-
Drug—O O Drug—OH +
Drug—O
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
OH O
F O S
F
O
Cl N O
O Cl N
H2N O
H2N
oxindole
8.1 8.2
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
O O
P
O

8.4
 Amine Prodrugs
Table 8.2
Drug—NH 2 Drug—NHX

O O O O
+
CR CCHNH3 C OPh —CH 2NHCAr CHAr NAr
R

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
O
N
F NH2

hydrolyze imine and

amide to GABA
Cl inside brain
progabide
8.6
anticonvulsant
 A Reductive Carrier-Linked Prodrug
Approach
Scheme 8.3
OH OH

NO2 :NH N
Y Y XH Y
bioreduction Drug +
X Z X Z Z
Drug Drug
8.7 8.8
 Prodrugs of Sulfonamides
A water soluble prodrug of the anti-inflammatory
drug valdecoxib (8.9) has been made (8.10).
Ph N Ph N
O O
Na+
H 2N CH3 N CH3
S S
O O O O O

valdecoxib parecoxib sodium

8.9 8.10
 Prodrug Analogs of
Carbonyl Compounds
Table 8.3
Drug Drug X
C=O C
R R Y

X
C
Y

OR' O S
C=NR' C=NOH C C C
OR' N N
H H

imines oximes ketals

Examples of Carrier-Linked
Bipartate Prodrugs
 O
Prodrug for Increased
R' = CCH2CH2CO2 Na
Water Solubility
OR'
Me OH Prodrug forms
HO
O
Me R' = PO3Na 2
for aqueous injection or
opthalmic use
O
R
prednisolone (R = R' = H) poor water
methylprednisolone (R = CH3 , R' = H) solubility
8.11
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.
O O
CH3 O
HO O
HO
O
O
O
O

CH3O OCH3
OR

etoposide (R = H)
etoposide phosphate (R = PO3 H2 )
8.12
Prodrug for Improved Absorption
Through Skin
OR
O
CH3 CH 3
HO O
CH3
CH3 O

F
O
F
fluocinolone acetonide (R = H)
fluocinonide (R = COCH3 )
8.14
corticosteroids - inflammation,
allergic, pruritic skin conditions
Better absorption into cornea for the treatment of
glaucoma
RO
OH
RO
NHCH3

dipivefrin (R = Me3 CCO)

epinephrine (R = H)
8.15

The cornea has significant esterase activity

Prodrug for Site Specificity
RO Bowel sterilant
OR

O
N
H
oxyphenisatin (R = H) (administer rectally)
8.16

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 NH2
O
O
NH2
O
8.17
 OR
Site Specificity
Using Enzymes at
the Site of Action

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:
ADEPT
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
An example is carboxypeptidase G2 or alkaline phosphatase
linked to an antibody to activate a nitrogen mustard prodrug.
Scheme 8.4
I I I
I
N N

carboxypeptidase G2
+ L-Glu + CO 2

H
O N CO2H OH

O CO2H
electron-donating;
8.19 activates nitrogen mustard
electron-withdrawing;
deactivates 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 O OH O O OH O
OH OH OH
OH 38C2 38C2
OH OH

CH3O O OH H O CH3O O OH H O B- O CH3O O OH H O

O
CH3 O retro-aldol O retro-Michael O
CH3 H CH3
NH O NH NH
HO HO O HO O
O H O O O
8.20 O O-
-CO2 +H+
B-

O OH O
OH
OH

CH3O O OH H O
CH3 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 Cl
OMe OMe

N nitroreductase H N OMe
OMe N
N HO N: H
O 2N O H O OMe
NMe OMe NMe

N N O NH
O NH -CO2
O O +H+

Cl
OMe

N OMe
N
O H
OMe

NH 2
Scheme 8.6 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 OR
F N

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 N H
H
N Cl
clindamycin (8.28) is
bitter and not well H O HO O
OH
tolerated by children. SCH3
OR
Clindamycin palmitate clindomycin (R = H)
is not bitter. clindomycin phosphate (R = PO 3H2)
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

H+
CH2O + NH3 N N
H3O+ N

methenamine
8.30
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
CH2 CH CH 2 CH
x y
OH O

O
O
8.32 O

Aspirin linked to poly(vinyl alcohol) has about the

same potency as aspirin, but less toxic.
 Steric Hindrance by Polymer Carrier
poly(methacrylate)
CH3 CH3
CH2 C CH2 C
x y
C=O C=O
O
O
to enhance
water
S=O solubility
O
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 CH2 C y
x
C=O C=O
O
O
to enhance
-
water
+ Cl
spacer NMe2 S=O solubility
arm
O H3 C

O
8.35
 Poly(α-Amino Acid) Carriers
O
poly(L-glutamine) NH CH C
x
spacer
O
O N O O
H C CH

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 for site specificity
8.38
Site-Specific Delivery of a Nitrogen Mustard
O O O
NHCH C NHCH C NHCH C poly(L-Glu)
x y z
spacer
O arm
O O
NH
O- NH

water-solubilizing Ig antibody from

N
rabbit antiserum
Cl Cl against mouse
lymphoma cells
8.39
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)
NH2
Tumor cells take up
proteins rapidly. Proteins N
broken down inside cells,
O N
releasing the drug. RO
O
HO
Shown to inhibit growth
of Ectomelia virus in OH
mouse liver, whereas free cytosine arabinoside (R = H)
inhibitor did not. 8.41
antitumor
 Antibody-Targeted Chemotherapy
Scheme 8.7 calicheamicin, except as disulfide
HO instead of trisulfide
O

NH O
CH3O
H polysaccharide HO
O
O
CH3 S
H O
N S NH
CH3O
N H polysaccharide
H RS-
N O O
antibody Lys O
S
O
gemtuzumab ozogamicin
8.42 8.43

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 —O- + RCOOH

fast

Drug—X- + CH 2O
 Tripartate Prodrugs of Ampicillin
O

Poor oral absorption (40%) NH

S
NH2
Excess antibiotic may destroy
N
important intestinal bacteria O
used in digestion and for COO-
ampicillin
biosynthesis of cofactors. 8.44
Also, more rapid onset of antibacterial
resistance.
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 O
Ph Ph
NH NH
S S
NH2 esterase NH2
+ R'COOH
N N
O O ..
O O R' O OH when
O O R' = OEt
R O R
8.46
bacampicillin (R = CH3 , R' = OEt) EtOH
pivampicillin (R = H, R' = t-Bu) O + CO 2
8.45
R H

98-99% absorbed 8.44

Ampicillin is released in < 15 minutes

 Reversible Redox Drug Delivery System to the CNS
Scheme 8.11 hydrolysis
hydrolysis deactivated activated
O O O

Drug—X crosses Drug—X enzyme Drug—X

blood-brain oxidation
N barrier N N+
CH3 CH3 CH 3
8.48
8.47
hydrophilic HOOC enzyme
hydrolysis
drug + Drug — XH
electron-
N+
electron-donating, CH3
withdrawing;
lipophilic carrier 8.49 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 self-
immolative reaction also can be used.
Scheme 8.12
O O O O
Drug C OCH2 O C carrier esterase Drug—C—O—CH2 —O- + HOC carrier

8.50 fast —CH2O

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.
H
R N S H
R N S
O
O N enzymatic O
O O N
O O oxidation O
O O O
N O
N
Me
8.51 Me
esterase
H H
R N S N O
R S
-CH2O
O N O N O
O O
O-
O O N
Scheme 8.13 O O Me
8.49
High concentrations of β-lactams delivered into brain.
 Tripartate Prodrug for Delivery of
Antibacterials
Permeases are bacterial transport proteins for uptake
of peptides.
O
Scheme 8.14 O
F
HN F
HN
O N +
1. permease H3N COO- O
O N
+ +
H3N 2. peptidase
CH3 ..
N COO- H2N COO-
H
CH3
O
8.53 H F
+ HN
H2N +
-
COO N
+ H O
NH 4 + H
O COO -

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 β-lactamase inactivator

ampicillin penicillanic acid sulfone
O
Ph
N O O
H S S
NH 2
N
O O O N
O O O

sultamicillin
8.59
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.

N NH2

8.60

Liver toxicity observed thought to be because of

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

Modification gave 8.62 with high activity.

N
S
N
N
8.62 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)
CH3
Modification of 8.63 gave O
N CO2CH3
8.64 having no iodine S
N
blockage activity. N
H
CH3
picoprazole
8.64
Picoprazole shown to be an
inhibitor of H+,K+-ATPase.
SAR of analogs indicated OCH3
CH3
electron-donating groups on H3 C O
N OCH3
the pyridine ring were S
N
favorable. Increased inhibition N
H
of H+,K+-ATPase. omeprazole
8.65
Best analog was 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.
OCH 3 OCH3 OCH3 OCH 3
H3 C CH3
Scheme 8.16 H3 C CH3 H3 C CH3 H3 C CH3

N -H2 O
.. N N N
S
S ..
O HN S HN S S
HN O OH N N
N N
N H
H+

OCH 3 OCH 3 OCH 3

OCH3
8.66
omeprazole
8.65
+H+

OCH 3
H3 C CH3

Formation of 8.66 leads to covalent attachment. N

Omeprazole also inhibits isozymes of carbonic N NH

S
S

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 R
Me S
O S Me
O- O O- S
S+ N N S+ N
S H N
Me
K2CO3/RX
HO O
O Me OH

Me HO O Me
HO O
leinamycin
8.70 8.71
antitumor agent prodrugs of leinamycin
HO- O Hydrolysis of these analogs gives an intermediate
O

Me
O
that reacts further to the activated form.
S- O Me S
S
O- Me S O O- Me S O-
O
S+ N N
S+ N S+ N S H
N N
Me Me
HO O HO O
O Me OH DNA
cleavage

Me O
Me Me O
O O O O

O O
O this was
8.73 synthesized
8.72

increased stability and gave

over leinamycin O
Me
S 8.74 as well
HO2 C
Me
N
H
N as DNA
HO S
cleavage
HO Me
Scheme 8.18 O O

8.74
 Elimination Activation
Scheme 8.19
CF 3 CF 3
B: O O
H N
N C
H N
H
N
O CH3
HO CH3
H
B 8.76 potent inhibitor of
leflunomide dihydroorotate
8.75
rheumatoid arthritis drug dehydrogenase

Inhibition of dihydroorotate dehydrogenase blocks

pyrimidine biosynthesis in human T lymphocytes.
 Oxidative Activation
N-Dealkylation
Sedative 8.20
CH 3 CH3 CH 3
N N N

N N N N N
N
CH3 P450 CH3 P450 NH
.. 2
O N N O
Cl O
CH 3 Cl H Cl
X X X

CH 3 N CH3 N
N
N
N
-H2 O N

Cl N ..
Cl NH
HO X
X

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.
HO:
H H O
H NH
Scheme 8.21 N O P-450 N O 2
O
P Cl P Cl P
H Cl
O N O N O N
B:
Cl Cl Cl
cyclophosphamide 8.80 8.81
8.79

H2N O
- P O
O Cl
Cl spontaneous N H
HPO4= + NH3 + HN or +
phosphoramidase Cl
Cl 8.83
8.82
Cyclophosphamide was very 8.84

effective, but it required liver DNA DNA

homogenates (contains P450) O

H
N O
H
N
+ OH OH
N N
for activation. Therefore HN HN

H2N N N H 2N N N
oxidation is required, not DNA
8.86
hydrolysis. 8.85
isolated
 N-Oxidation
identified
O O O
P450 -H2 O
CH3 NHNHCH 2 CNHCHMe 2 CH3 NHNHCH 2 CNHCHMe 2 CH3N=N—CH CNHCHMe 2
+
HO B—H H
procarbazine 8.88
8.87 B:

advanced Hodgkin’s
disease O O
H 2O
CH 3N-NH2 + OHC CNHCHMe 2 CH3N-N=CH CNHCHMe 2
H H

8.89

CH3N=NH
O CH3 O CH3
+ +N
CH3-N N HN HN N
DNA

CH3 + N2 H 2N N N H2N N N
DNA
8.90
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.

N OH
+N
Cl -
CH3
pralidoxime chloride
8.91
 To increase the permeability of pralidoxime into
the CNS, the pyridinium ring was reduced (8.92).
N OH
N
CH3
8.92

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 Me 3N—CH2CH2—OH
Trp
H CH3 Trp
Trp Trp O +
Phe B+ :B B: HB
H Phe CH3
O O

H2O

+
Me 3NCH2CH2OH + CH3 COOH
 Inactivation of Acetylcholinesterase by
Diisopropyl Phosphorofluoridate
Scheme 8.24
affinity labeling agent
O
O O O
F P
Trp P
H O Trp O
Phe Trp B+ :B O
H Phe Trp B HB
O

8.93
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 O H O
N O
P N O:
pralidoxime P
Trp O Trp Me O
O O
Phe Trp B HB Phe Trp B HB

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

neostigmine covalent, but

8.94
reversible slow H 2O
Used for the neuromuscular
inhibition
disease myasthenia gravis
+ CO 2 + Me2NH
Me 3N OH
 Reversible (noncovalent) inhibitors of
acetylcholinesterase, such as donepezil and tacrine
are used for Alzheimer’s disease. Enhances
neurotransmission involved in memory.
O
H3CO NH2

H3CO
N
N.
. HCl
HCl
donepezil hydrochloride tacrine hydrochloride
8.95 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
:B
H HO H HO H HO H H
O O
HO RSH [O] O
O CH3 HO RS H HO RS
O CH3 H O CH3
H H O
H
brefeldin A
8.97 8.98
8.99

-RSOH

antitumor,
antiviral agent
 Aromatic Hydroxylation
Cyclohexenones as prodrugs for catechols
Scheme 8.28

N -H2 O N N
P450 N

HO
O O OH
O
P450
8.100 aromatic hydroxylation
oxidation next to
sp2 carbonyl N

HO
OH
8.101
 Alkene Epoxidation

N
N
O NH2
O 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 OH Anaerobic cleavage

N
by bacteria in lower
sulfasalazine
8.106 bowel
ulcerative colitis COOH

H 2N OH + NHSO2 NH2
N

8.107 sulfapyridine
8.108
For inflammatory Causes side effects
bowel disease
To prevent side effect by sulfapyridine a
macromolecular delivery system was developed.
n poly(vinylamine)
NH
SO2
Not absorbed or
metabolized in small
spacer CO2Na
intestine.
N N OH

8.109
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
N

N N
HO
O
OH

HO
vidarabine (R = NH 2)
8.110
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
CO2H
F
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.

CO2H CO2H
F MeO
CH3 CH3
N NH2

O HO

CH3 S N
Cl
O H
indomethacin serotonin
sulindac 8.112
8.111
 Disulfide Reduction
To increase the lipophilicity of thiamin for absorption
into the CNS.
OH OH
Scheme 8.30 ..
N GSH N
N S S O N S-
O H O H
CH3 N NH2 CH3 N NH2
+B H
8.113

nonenzymatic

OH
+ N:
N N OH N S
S
CH3 N NH2 CH3 N NH2 OH

thiamin
8.114
poorly absorbed into CNS
CH3O
To diminish toxicity of primaquine
N and target it for cells with the malaria
HN parasite, a macromolecular drug
NH2

delivery system was designed.

primaquine
8.115
antimalarial, toxic Intracellular thiol much higher
than in blood; selective
CH3O reduction inside the cell
lactose

N O
HN serum
N S S
H albumin
NH3+

primaquine 8.116 for lactose

improved
uptake in liver

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

 Nitro Reduction
Scheme 8.32
O O O
F F Cl
HN HN F
O O HN
Cl Cl O
O bioreduction O ..
N P O N N P O N N P O O N
H3 C O O H3 C O O
O- O
H3 C

HO HO
O O HO
8.120
..
O 2N HO NH
8.118 8.119

O O O
F F F
HN HN HN
O O
O
O H 2O P O O N N P O O N
-O
P O N
O O O H3 C O- O
O-

HO HO HO
8.121

Mechanism-based inactivator of
thymidylate synthase
 Nucleotide Activation
Scheme 8.33
= SH
O3PO O O O
SH N N
O—P—O—P—O-
N N N N
HO OH O -
O - =O
3PO O
N N hypoxanthine-guanine
H phosphoribosyltransferase
6-mercaptopurine HO OH
8.122
Anti-leukemia drug 8.123
Inhibits several enzymes in
the purine nucleotide
biosynthesis pathway.
 Phosphorylation Activation
O
O N
HN
HN N
H 2N N N
H2N N N HO
RO O
O
HO
acyclovir (R = H)
8.124 8.125
antiviral 2′-deoxyguanosine
Resembles structure of 2′-deoxyguanosine
viral thymidine kinase
Uninfected cells do not R = PO3=
phosphorylate acyclovir viral guanylate kinase
(selective toxicity) 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 NH2
here Prodrug for
N
N
a prodrug
H2N N N
HO
O

8.126
adenosine deaminase

acyclovir
 Hydroxylates
here
N
N

H 2N N N
HO
O

8.127

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 N

H 2N H 2N N N
O
O
O

valaciclovir
8.128
An analog of acyclovir whose structure is even
closer to that of 2′-deoxyguanosine is ganciclovir.
O

HN N

H 2N N N
HO
O

HO
ganciclovir
8.129

More potent than acyclovir against human

cytomegalovirus.
Two carbon isosteres of ganciclovir are available.
O
N N
HN N

H 2N N N H 2N N N
HO AcO

C in place AcO
C in place
HO
of O of O
penciclovir famciclovir
8.130 8.131

Better oral absorption

than penciclovir
Converted to penciclovir
rapidly
 Sulfation Activation
Activity of minoxidil requires sulfotransferase-catalyzed
sulfation to minoxidil sulfate.
OR
H2N 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 blood- L-aromatic amino acid

brain barrier 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 OH
HO O NH2
+
NHNH3 HO
H3 C N N OH
COO- H H
carbidopa benserazide
8.134 8.135
(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
decarboxylation outside of the brain. F

NH2

OH
8.136
 Selective Delivery of Dopamine to Kidneys
Dopamine increases renal blood flow.
Prodrugs were designed for selective renal vasodilation.
Scheme 8.34 OH
OH
COO- L-γ-glutamyl
H transpeptidase
+ N OH + OH
NH 3 NH 3
O COO - COO-
Glu
8.137 L-dopa
L-γ-glutamyl-L-dopa L-aromatic amino acid
decarboxylase

+
Dopamine accumulates in the kidneys because of NH3 OH
high concentrations of L-γ-glutamyltranspeptidase
and L-aromatic amino acid decarboxylase there. OH

Example of a carrier-linked prodrug of a bioprecursor

prodrug for dopamine.