“REDUCTION OF CARBONYL COMPOUND

BY LiAlH4 & NaBH4”

Presentation submitted in the seminar of 2nd semester

(2019-20).
MASTER OF PHARMACY
IN
PHARMACETICAL CHEMISTRY

By
Panchal Himanshu Nilesh

Guided By
Dr.Prashant Murumkar

FACULTY OF PHARMACY
THE MAHARAJA SAYAJIRAO UNIVERSITY
OF BARODA
VADODARA- 390002, GUJARAT.
 CERTIFICATE
This is to certify that the seminar entitled “REDUCTION OF
CARBONYL COMPOUNDS USING EMMENSEN AND
WOLFF ISHNER REDUCTION” has been presented by
PANCHAL HIMANSHU to faculty of Pharmacy at THE
MAHARAJA SAYAJI RAO UNIVERSITY OF BARODA, under
my guidance.

GUIDE HEAD OF THE DEPARTMENT

Dr. Prashant Murumkar Prof. K.K. Sawant
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1. Introduction
• Almost all drugs possess some undesirable physicochemical and biological properties.
• The concept of prodrug is first introduced by Adrian Albert in 1958. He stated that
“Compounds that undergo biotransformation prior to eliciting their
pharmacological effect are the prodrugs.”
• ”A prodrug is any compound that undergoes biotransformation before exhibiting
its pharmacological effects” definition of prodrug given by IUPAC.
• Prodrug is pharmacologically inactive compound. Generally, the metabolic
transformation is necessary to convert the prodrug into its active form which is
catalyzed by specific enzymes mainly hydrolases and ideally this should selectively
occur at the target tissue to prevent undesirable side effects.
• The development of prodrugs is now well established as a strategy to improve the
physicochemical, biopharmaceutical or pharmacokinetic properties of
pharmacologically potent compounds and thereby increase usefulness of a potential
drug. This process is schematically represented in fig.1.1.

(Active drug)

Figure.1.1: Schematic representation of biotransformation process of activation of

prodrug.

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2. History
• Many herbal extracts historically used in medicine contain glycosides (sugar
derivatives) of the active agent, which are hydrolyzed in the intestines to release the
active and more bioavailable aglycone. For example, salicin is a β-D-
glucopyranoside that is cleaved by esterase to release salicylic acid.
• Aspirin acetylsalicylic acid first made by Felix Hoffmann at Bayer in 1897 is a
synthetic prodrug of salicylic acid.
• However, in other cases such as Codeine and Morphine the administered drug
is enzymatically activated to form sugar derivatives (morphine-glucuronides) that are
more active than the parent compound.
• The first synthetic antimicrobial drug Arsphenamine, discovered in 1909
by Sahachiro Hata in the laboratory of Paul Ehrlich is not toxic to bacteria until it has
been converted to an active form by the body.
• Likewise, Prontosil the first sulfa drug (discovered by Gerhard Domagk in 1932),
must be cleaved in the body to release the active molecule sulfanilamide. Since that
time many other examples have been identified.
• Terfenadine the first non-sedating antihistamine had to be withdrawn from the market
because of the small risk of a serious side effect.
• However, Terfenadine was discovered to be the prodrug of the active
molecule Fexofenadine which does not carry the same risks as the parent compound.
• Therefore, Fexofenadine could be placed on the market as a safe replacement for the
original drug. Loratadine another non-sedating antihistamine is the prodrug
of desloratadine which is largely responsible for the antihistaminergic effects of the
parent compound. However, in this case the parent compound does not have the side
effects associated with terfenadine and so both loratadine and its active metabolite
desloratadine are currently marketed.

3. Rationales of prodrug
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1. Aqueous Solubility
2. Absorption and Distribution
3. Site Specificity
4. Instability
5. Prolonged Release
6. Toxicity
7. Poor Patient Acceptability
8. Formulation Problems

4. Types of prodrug
• There are mainly 2 types of prodrugs.
1. Carrier-Linked Prodrugs
-Bipartite prodrug
-Tripartite prodrug
-Mutual Prodrug
-Macromolecular Drug Carrier Systems
2. Bioprecursors prodrug

1.4.1 Carrier-Linked Prodrugs-

➢ A carrier-linked prodrug is a compound that contains an active drug linked to a carrier
group that can be removed enzymatically such as an ester which is hydrolyzed to an
active carboxylic acid-containing drug.
➢ A bipartite prodrug is a prodrug composed of one carrier attached to the drug.
➢ When a carrier is connected to a linker that is connected to the drug it is called a
tripartite prodrug.
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➢ A mutual prodrug (also known as a co-drug) consists of two usually synergistic drugs
attached to each other (one drug is the carrier for the other and vice versa).

Figure 1.2: Carrier-Linked Prodrugs.

➢ In Macromolecular Drug Carrier Systems drug is covalently attached to a

macromolecule such as a synthetic polymer a glycoprotein, a lipoprotein, a lectin, a
hormone, albumin, a liposome, DNA, dextran, an antibody or a cell.
➢ The pharmacokinetic characteristics of these drugs change dramatically which
depends on the physicochemical properties of the macromolecular carrier not the
drug.
➢ Several examples of macromolecular drug carrier systems follow.
a) Synthetic Polymers
b) Poly(α-Amino Acids)

1.4.2 Bio- precursor prodrugs.

➢ Produce their effects after in vivo chemical modification of their inactive form.
➢ Bioprecursor prodrugs rely on oxidative or reductive activation reactions unlike the
hydrolytic activation of carrier-linked prodrugs.
➢ They metabolized into a new compound that may itself be active or further
metabolized to an active metabolite.
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➢ Type of metabolic activation reaction involved. The first example is the simplest of
metabolic transformations, namely, protonation as a mechanism for prodrug
activation
• Protonation activation
• Hydrolytic Activation
• Elimination Activation
• Oxidative Activation
❖ N- and O-Dealkylations
❖ Oxidative Deamination
❖ N-Oxidation
❖ S-Oxidation
❖ Aromatic Hydroxylation
❖ Other Oxidations
• Reductive Activation
❖ Azo Reduction
❖ Azido Reduction
❖ Sulfoxide Reduction
❖ Disulfide Reduction
❖ Nitro Reduction
• Nucleotide Activation
• Phosphorylation Activation
• Sulfation Activation
• Decarboxylation Activation

1.4.3. Functional Groups Amenable to Prodrug Design.

1. Esters as prodrugs of carboxyl, hydroxyl and thiol functionalities

➢ Esters are the most common prodrugs used and it is estimated that approximately
49% of all marketed prodrugs are activated by enzymatic hydrolysis.
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➢ Ester prodrugs are most often used to enhance the lipophilicity and thus the passive
membrane permeability of water soluble drugs by masking charged groups such as
carboxylic acids and phosphates.
➢ The synthesis of an ester prodrug is often straightforward. Once in the body the ester
bond is readily hydrolysed by ubiquitous esterases found in the blood, liver and other
organs and tissues including carboxyl esterases, acetylcholinesterases,
butyrylcholinesterases, paraoxonases and arylesterases.

2. Carbonates and carbamates as prodrugs of carboxyl, hydroxyl or amine

functionalities.
➢ Carbonates and carbamates differ from esters by the presence of an oxygen or
nitrogen on both sides of the carbonyl carbon.
➢ They are often enzymatically more stable than the corresponding esters but are more
susceptible to hydrolysis than amides.
➢ Carbonates are derivatives of carboxylic acids and alcohols while carbamates are
carboxylic acid and amine derivatives.
➢ The bioconversion of many carbonate and carbamate prodrugs requires esterases for
the formation of the parent drug.

3. Amides as prodrugs of carboxylic acids and amines

➢ Amides are derivatives of amine and carboxyl functionalities of a molecule. In
prodrug design amides have been used only to a limited extent owing to their
relatively high enzymatic stability in vivo.
➢ An amide bond is usually hydrolyzed by ubiquitous carboxylesterases, peptidases or
proteases. Amides are often designed for enhanced oral absorption.
➢ Lipophilicity of various compounds like acid chlorides and acids can be altered in
many groups of compounds by amide formation.
➢ This approach is successful to improve the stability of drug in vivo in many of the
pharmaceutical agents and gives targeted drug delivery due to presence of enzyme
amydase.
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4. Oximes as derivatives of ketone, amidine and guanidine

➢ Oximes (for example, ketoxime, amidoxime and guanidoximes) are derivatives of
ketone, amidines and guanidine thus providing an opportunity to modify molecules
that lack hydroxyl, amine or carboxyl functionalities.
➢ Oximes are hydrolyzed by the versatile microsomal cytochrome P450 (CYP450)
enzymes.
➢ Oximes especially strongly basic amidines and guanidoximes can be used to enhance
the membrane permeability and absorption of a parent drug.

5. Examples of several types of prodrug

1.5.1. Bipartite prodrug
➢ As oncostatic drugs are endowed with poor selectivity. The lack of selectivity of
anticancer drugs and associated toxicity hampers their effectiveness and long term
use. Hence, not surprisingly there is an urgent need to improve their selectivity.
➢ Prodrug technology can be used to site specific delivery of anticancer drugs.
➢ Anticancer prodrugs can be designed to target specific molecules (enzymes, peptide
transporters, antigens) that are overexpressed in tumor cells in comparison to normal
cells. The new promising chemotherapeutic prodrugs include:
a) Enzyme-activated prodrugs.
• Antibody-directed enzyme prodrug therapy (ADEPT).
• Gene-directed enzyme prodrug therapy (GDEPT).
• Virus-directed enzyme prodrug therapy (VDEPT).
b) Targeting-ligand conjugated prodrugs
• Antibody-drug conjugates
• Peptide-drug conjugates

Eg. EIDD 2801

EIDD-2801 is the isopropylester prodrug of N4-hydroxycytidine.With improved oral

bioavailability in non human primates, it is hydrolyzed in vivo, and distributes into tissues
where it becomes the active 5’-triphosphate form. The active drug incorporates into the
genome of RNA viruses, leading to an accumulation of mutations known as viral error
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catastrophe. Recent studies have shown EIDD-2801 inhibits replication of human and bat
corona viruses, including SARS-CoV-2, in mice and human airway epithelial cells.

Figure- 1.3: EIDD 2801

Figure- 1.4: N4-hydroxycytidine

1.5.2. Tripartate prodrug

➢ Redox drug delivery system is the best example of tripartite prodrug system.
➢ The blood–brain barrier is an important membrane for protection of the brain from
polar, hydrophilic molecules that do not belong there.
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Figure1.5: Mechanism of active drug release from tripartate prodrug.

Figure 1.6: Brentuximab vedotin, the first antibody-drug conjugates with a more
stable linker.

Figure-1.7- Redox drug delivery system in brain

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➢ Reversible redox drug delivery system for getting drugs into the central nervous
system and then once in from preventing their passive efflux
➢ Furthermore, the nitrogen atom in the dihydropyridine ring is conjugated.
➢ Through the double bond into the carbonyl (a vinylogous amide) thereby making the
carbonyl less reactive toward nucleophiles such as water and therefore more stable to
hydrolysis.
➢ Once inside the brain, the lipophilic carrier is converted enzymatically into a highly
hydrophilic species in which the pyridinium nitrogen atom is no longer conjugated
into the carbonyl in fact, the pyridinium group is electron withdrawing so it now
activates the carbonyl for nucleophilic attack.
➢ The drug is readily released by enzymatic hydrolysis and the N-methylnicotinic acid
is relatively nontoxic and actively transported out of the brain.
➢ The XH group on the drug is an amino, hydroxyl or carboxyl group. When it is a
carboxylic acid the linkage is an acyloxymethyl ester which decomposes by the self-
immolative.
➢ The oxidation of the dihydropyridine to the pyridinium ion (half-life generally 20–50
min) prevents the drug from escaping out of the brain because it becomes charged.
This drives the equilibrium of the lipophilic precursor throughout all the tissues of the
body to favor the brain. Any oxidation occurring outside the brain produces a
hydrophilic species that can be rapidly eliminated from the body.

1.5.3. Macromolecular Drug Carrier Systems

• Synthetic polymers-
➢ Aspirin linked to poly (vinyl alcohol) (figure-1.8.) was shown to have the same
potency as aspirin, but was less toxic. Another anti-inflammatory agent, ibuprofen
(the carboxylic acid from hydrolysis, Advil), was attached as a poly (oxyethylene)
diester. This macromolecular carrier system resulted in a sustained release of
ibuprofen, giving prolonged anti-inflammatory activity and a higher plasma half-life
relative to the free drug.
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CH 2 CH CH 2 CH
OH

O O

Figure-1.8: Aspirin linked to poly (vinyl alcohol).

➢ Because it is necessary for the drug to be released from the polymer backbone, steric
hindrance by the polymer to chemical or enzymatic hydrolysis may cause problems.
To avoid steric hindrance by the polymer in the first synthetic step, a spacer was
incorporated between the polymer and the first building block.
• Alpha amino acid
➢ The disadvantage of using synthetic polymers is that they are generally not
biodegradable, and they can take 5–12 months to be eliminated from the body.
Poly(α-amino acids) are biodegradable (at least the l-isomers are); the rate of
biodegradability depends on the choice of amino acid
➢ Conjugation of the antitumor drug methotrexate (Rheumatrex) to poly(l-lysine)
(figure-1.8.); attachment of the polymer also may be to the α-carboxyl group of the
terminal lysine residue) markedly increased the cellular uptake of the drug and
provided a new way to overcome drug resistance related to deficient drug transport.
Because the activity of methotrexate is a function of its inhibitory properties of
dihydrofolate reductase and fig-1.9. is a poor inhibitor of this enzyme in vitro, the
methotrexate must become detached from the polymer backbone inside the cell.
Furthermore, attachment of methotrexate to poly(d-lysine), which, unlike poly(l-
lysine) does not undergo proteolytic digestion inside the cell, gave a conjugate devoid
of activity with resistant or normal cell lines. Methotrexate attached to poly(l-lysine)
is also more inhibitory to the growth of human solid tumor cell lines than to the
growth of human lymphocytes; free methotrexate is equally toxic to both kinds of
cells.
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O
NH CH C

HOOC N N NH2
HN
NH
N
O N N
NH2

Figure-1.9: Methotrexate attached to poly(l-lysine).

1.5.4. Mutual prodrug-
➢ When it is necessary for two synergistic drugs to be at the same site at the same time,
a mutual prodrug approach should be considered. A mutual prodrug (or codrug) is a
bipartite or tripartite prodrug in which the carrier is a synergistic drug with the drug to
which it is linked. The problems that arise from this type of prodrug relate to the
much larger molecular weight of the combined drugs and the associated side effects
from each.
➢ For example, the combination of the penicillin derivative amoxicillin (2.0.; Amoxil)
and the β-lactamase inactivator potassium clavulanate (2.1.) (the combination is
called Augmentin) is used for oral treatment of infections caused by β-lactamase-
producing bacteria.
O

NH S
HO
NH2
N
COOH
Figure-2.0. Amoxicillin

OH

N
O
COOK

Figure-2.1. Potassium clavulanate

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1.5.5. Bioprecursor Prodrugs

A) Proton Activation-
➢ Antiulcer drugs cimetidine and ranitidine These compounds lowered gastric acid
secretion by antagonizing the H2 histamine receptor.
➢ Another approach for lowering gastric acid secretion is by inhibition of the enzyme
H+,K+- ATPase (also known as the proton pump), which is responsible for acid
secretion by the parietal cell.

Figure-2.2. Mechanism of inactivation of H+,K+-ATPase by omeprazole

B). Oxidative Activation

a) N- and O-Dealkylations
➢ Open ring analogs of benzodiazepines such as the anxiolyticdrug alprazolam (2.1.1, X
= H; Xanax) and the sedative triazolam (2.1.1, X = Cl; Halcion), undergo metabolic
N-dealkylati and spontaneous cyclization.
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Figure 2.3: Bioprecursor prodrugs for alprazolam and triazolam

b) Oxidative Deamination

➢ Because of the high concentration of phosphoramidases in neoplastic cells, hundreds

of phosphamide analogs of nitrogen mustards were synthesized and tested as
carrierlinked antitumor prodrugs. Cyclophosphamide (1 in figure 2.4 ; Cytoxan)
emerged as an important drug for the treatment of a wide variety of malignant
diseases; however, it was later found that it was inactive in tissue culture, suggesting
that simple hydrolysis was not involved. Preincubation of the compound with liver
homogenates, however, activated it, suggesting that cyclophosphamide is a prodrug
requiring an oxidative mechanism.
➢ The activation mechanism is believed to be that shown in figure 2.4 (there are other
metabolites that are not shown in this scheme derived from each of the intermediates).
It is not clear which of the toxic metabolites, the phosphoramide mustard (4) or the
parent nitrogen mustard (6), is responsible for the therapeutic action; the major adduct
isolated by high-performance liquid chromatography from in vitro and in vivo studies
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in rat is N-(2-hydroxyethyl)-N-[2-(7-guaninyl)ethyl]amine.(8), the hydrolysis product

of (7). Nitrogen mustard reacts with DNA.
➢ Acrolein (5) is a potent Michael acceptor that may be responsible for the
hemorrhagic cystitis side effect; administration of sulfhydryl compounds, which react
readily with acrolein, can prevent this side effect.

Figure- 2.5. Cytochrome P450-catalyzed activation of cyclophosphamide.

c) Reductive Activation

a) Azo Reduction-

➢ Prontosil, is activated by reduction of its azo linkage to the true bacteriostatic agent,
sulfanilamide. Sulfasalazine (1; Azulfidine), which is used in the treatment of
inflammatory bowel disease (ulcerative colitis) and rheumatoid arthritis is reductively
cleaved by anaerobic bacteria in the lower bowel to 5-aminosalicylic acid (2) and
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sulfapyridine (3) (figure-2.6); (2) is the therapeutic agent, and produces adverse side
effects.
➢ A macromolecular drug delivery system was developed to improve the therapeutic
index of this
drug. The drug (2) was azo-linked at the 5-position through a spacer to poly(vinyl
amine) to give figure 2.7.
➢ The advantages of this polymeric drug delivery system are that it is not absorbed or
metabolized in the small intestine, (2) can be released by reduction at the disease site,
and the carrier polymer is not absorbed or metabolized.
➢ The water-soluble polymer-linked drug fig.2.7 was more potent than (1) or (2) in the
guinea pig ulcerative colitis model.

Figure 2.6: Reductive activation of sulfasalazine

n
NH

O 2S

COONa
N
N OH

Figure 2.7: polymer linked sulfasalazine.

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b) Sulfoxide Reduction
➢ The antiarthritis drug sulindac (2.8; Clinoril) is an indene isostere of the nonsteroidal
antiinflammatory (antiarthritis) drug indomethacin (2.9; Indocin), which originally
was designed as a serotonin analog. Sulindac is less irritating to the gastrointestinal
tract and produces many fewer and more mild central nervous system effects than
does indomethacin.
➢ The 5-fluoro group was substituted for the methoxyl group to improve the analgesic
properties, and the p-methylsulfinyl group was substituted for the chlorine atom to
increase the solubility. Sulfoxide Reduction The antiarthritis drug sulindac (2.8;
Clinoril) is an indene isostere of the nonsteroidal antiinflammatory (antiarthritis) drug
indomethacin (2.9; Indocin), which originally was designed as a serotonin analog.
Sulindac is less irritating to the gastrointestinal tract and produces many fewer and
more mild central nervous system effects than does indomethacin.. The 5-fluoro
group was substituted for the methoxyl group to improve the analgesic properties, and
the p-methylsulfinyl group was substituted for the chlorine atom to increase the
solubility.

COOH
F COOH

CH 3 MeO
CH3
N
O
H3C
S
O Cl

Figure 2.8: Clinoril Figure 2.9: Indocin

C) Nucleotide Activation.
➢ The antineoplastic agent 6-mercaptopurine (fig-3.0 (1); Purinethol) produces a 50%
remission rate for acute childhood leukemias. Although (1) inhibits several enzyme
systems, these inhibitions are irrelevant to its anticancer activity. Only tumors that
convert the drug to its nucleotide are affected. 6-Mercaptopurine is activated.
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➢ By a hypoxanthine-guanine phosphoribosyltransferasecatalyzed reaction with 5

phosphoribosylpyrophosphate (figure-3.), The nucleotide (2) inhibits several enzymes
in the purine nucleotide biosynthetic pathway, but the most prominent site is one of
the early enzymes in the de novo pathway, namely, phosphoribosylpyrophosphate
amidotransferase, which catalyzes the conversion of phosphoribosylpyrophosphate to
phosphoribosylamine.
➢ 5-Fluorouracil is similar to 6-mercaptopurine in the sense that it must first be
converted to the corresponding deoxyribonucleotide for it to be active.

Figure 3: Nucleotide activation of 6-mercaptopurine.

Applications

1. To improve aqueous solubility

➢ When our targeted dosage form is injectable or drops and also drug is insoluble in
water that it would need to be taken up in more than a liter of saline to administer the
appropriate dose.
➢ These drugs could be safe, effective and potent but they would not be viable for their
applications. Hence, water solubilizing group can be attached to the drugs which can
metabolically released after drug administration.

2. To improve Absorption and Distribution.

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➢ If drugs have problem with absorption from administration site and distribution from
administration site to the target site.
➢ It can be made more water soluble or lipid soluble depending on the desired site of
action by attaching water soluble or lipid soluble group with drug that attached
group will be removed enzymatically in the body.

3. To make site specific drug.

➢ Target specific drugs can be made for any organ when there is high concentration of
particular enzyme is present. Hence, it can cleave the produg at the site of interest and
covert in to active drug.

4. To improve patient acceptability.

➢ An active drug may have an unpleasant taste or odor, produce gastric irritation, or
cause pain when administered (for example, when injected). The structure of the drug
could be modified to alleviate these problems but once administered the prodrug
should be metabolized to the active drug.

5. To solve formulation problem.

➢ If the drug is a volatile liquid it would be more desirable to have it in a solid form so
that it could be formulated as a tablet. An inactive solid derivative could be prepared
which would be converted in the body to the active drug.

6. Improve stability.
➢ A drug may be rapidly metabolized and rendered inactive before it reaches the site of
action. The structure may be modified to block that metabolism until the drug is at the
desired site.

7. Prolonged Release
➢ It may be desirable to have a steady low concentration of a drug released over a long
period of time. The drug may be altered so that it is metabolically converted to the
active form slowly.
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8. To decrease Toxicity
➢ A drug may be toxic in its active form and would have a greater therapeutic index if it
were administered in a nontoxic inactive form that was converted into the active form
only at the site of action.
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