 Oxidative – The most common pathway for

PHASE 1 (FUNCTIONALIZATION): METABOLISM

another Phase I.
Metabolism  Polar Functional groups
1. OH
 plays a central role in the elimination of drugs and other 2. COOH
foreign compounds (xenobiotics) from the body. 3. NH2
 an essential tool for pharmacists in their role of 4. SH
selecting and monitoring appropriate drug therapy for
their patients. R ROH RRCOOH
 occur at some point between absorption into the
circulation and its renal elimination. R RSH R RNH₂
BIOTRANSFORMATION
 By direct introduction of the functional group
INACTIVE
 Example: aromatic and aliphatic hydroxylation
(prodrug)
 By modifying or unmasking existing functionalities
ACTIVE
(prodrug)  Examples:
PARENT
DRUG reduction of ketones and aldehydes to alcohols
NON - TOXIC
oxidation of alcohols to acids
hydrolysis of ester and amides to yield COOH, NH2,
TOXIC
and OH groups; reduction of azo and nitro compounds
 Inactive – Parent compound must be converted to to give NH2 moieties; oxidative N-, O-, and S-
Active. dealkylation to give NH2, OH and SH groups.
PHASE I REACTION
REACTIONS UNDER PHASE I
Goals
1. Oxidation
- Produce a more water-soluble compound.
- Most are mediated by microsomes.
- Produce a molecule that can undergo subsequent phase
- Although oxidation would seem to imply the addition of
II reactions.
oxygen, that is not always the case.
- Oxidation refers to the change in oxidation state of the
substrate.

2 TYPES OF OXIDATIONS:

1. Oxygen is incorporated into the drug molecule (e.g.

hydroxylation)
2. Oxidation causes the loss of part of the drug molecule
(e.g. oxidative deamination, dealkylation).
1. LIVER  Flavoprotein, NADPH-cytochrome reductase.
 Main site of metabolism and detoxification of  One mole of this enzyme contains one mole each of
endo/exogenous compounds flavin mononucleotide (FMN) and flavin adenine
 Rich in almost all the drug-metabolizing enzymes. dinucleotide (FAD).
 Other sites include the gut, lungs, skin, and kidneys.  Enzyme is also called NADPH-cytochrome P450
2. PO DRUGS reductase.
 Pass through the liver before being further distributed  Electron transporter
in body compartments.  Cytochrome P450
3. FIRST-PASS EFFECT  named based on its light absorption at 450 nm
 Metabolism before reaching systemic circulation. when complexed with carbon monoxide.
 Limit the BA of orally administered drugs.  is a hemoprotein containing an iron atom that can
 Intestinal metabolism alternate between the ferrous (Fe++) and ferric
 Liver metabolism (Fe+++) states.
 Enterohepatic recycling  Electron acceptor.
 Gut microorganisms - glucuronidases  No substrate specificity.
4. EXTRAHEPATIC METABOLISM 
 Intestine, kidney, lungs, adrenal glands, placenta,
brain, skin. HUMAN LIVER P450 enzymes
 Substrate specific  CYP3A4

Phase I (Functionalization)

 polar functional groups are introduced into the molecule

or unmasked by:
  Responsible for metabolism of over 50% of
prescription drugs metabolized by the liver.
Aromatic hydroxylation
CYP isoform Substrates - refers to the mixed-function oxidation of aromatic
3A4 APAP. Erythromycin, Lidocaine, Lovastatin, compounds (arenes) to their corresponding phenolic
Miconazole, Quinidine, Verapamil metabolites (arenols).
2D6 Clozapine, Codeine, Desipramine, - proceed initially through an epoxide intermediate called
Dextromethorphan, Fluoxetine, Haloperidol, an "arene oxide.” which rearranges rapidly and
Oxycodone, Paroxetine, Propafenone spontaneously to the arenol.
2C8 Taxol, Retinoic Acid Derivatives
2C9 Celecoxib, Diclofenac, Ibuprofen, Losartan,
Phenytoin, Tolbutamide
Microsomal Mixed Function Oxidases (MFQOs)

 “Microsomes” form in vitro after cell homogenization

and fractionation of ER
 Rough microsomes are primarily associated with protein  NIH Shift leading to the
synthesis. formation of arenols.
 Smooth microsomes contain a class of oxidative  May also acted upon by
enzymes called “Mixed Function Oxidases” or epoxide hydrases
“Monooxygenases.” leading to the formation
of trans-dihydrodiols.
MFO Requirements  Or be acted upon by
- require a reducing agent. (Electron donor) sulfhydryl group in GSH
1. NADPH (Nicotinamide Adenine Dinucleotide to yield GSH adduct.
Phosphate)  It is important to
2. Molecular oxygen prevent the formation of
- one oxygen atom appearing in the product. ROS by arene oxides.
- the other oxygen in the form of water.  Major route of metabolism for
Oxidative pathways phenyl-containing drugs.
- the cytochrome P-450 monooxygenase system plays a  Hydroxylation occurs at the
vital role in this biotransformation by: para position.
- Binding of the substrate (drug molecule, represented by  Microsomal aromatic
RH) to the oxidized (Fe+3) resting state of CYP-450 to hydroxylation reactions appear
form P-450- substrate complex. to proceed most readily in
- One electron from NADPH (nicotinamide-adenine activated (electron-rich) rings.
dinucleotide phosphate) is transferred to the P-450-  Drugs containing electron
substrate complex, which reduces Fe+3 to Fe+2. The P- withdrawing groups Cl, - NR3,
450-substrate complex is capable of binding dioxygen COOH, SO2NHR).
(O2).
- Dioxygen-P-450-substrate complex undergoes another Examples of drugs that undergo Aromatic Hydroxylation in
reduction to form peroxide dianion-P-450 (Fe+3)- humans.
substrate complex.
- Water containing one of the oxygen atoms is released to
form Activated oxygen (FeO)+3 that is highly electron-
deficient and a potent oxidizing agent.
- The activated oxygen is transferred to the substrate to
form ROH.

Cytochrome P450 cycle in drug oxidations

Examples of drugs resistant to Aromatic Hydroxylation in
humans

 Clonidine (Catapres)
 The
deactivating
groups (Cl. -N H
= C).
 Probenecid
(Benemid)
  electron-withdrawing groups: carboxy and Green Pigments
sulfamido group.  abnormal heme derivative
 N-alkylated protoporphyrins – Allylisopropyl
acetamide – Secobarbital – Fluoroxene

Compounds with two aromatic rings

 If there are two or

more phenyl rings,
hydroxylation
proceeds in the
electron-rich ring.
 There are certain
compounds that are resistant to aromatic hydroxylation
despite of the aromatic rings.
DUE TO presence of multi electronegative chlorine atoms
attached to the rings.
 Eg. Polychlorinated biphenyls (PCBs) 2,3,7,8-
tetrachlorodibenzo-p-dioxin

Oxidation of Olefins
Oxidation at Benzylic Carbon Atoms
 The metabolic oxidation of olefinic carbon—carbon
 Carbon atoms attached to aromatic rings (benzylic
double bonds leads to the corresponding epoxide (or
position) are susceptible to oxidation, forming the
oxirane).
alcohol (or carbinol) metabolite.
 The epoxide is cleaved by epoxide hydrases to form
 1⁰ alcohol metabolites are often oxidized further to
trans-diols.
aldehydes and carboxylic acids (CH2OH → CHO →
 There are inhibitors such as cyclohexene oxide and
COOH), and secondary alcohols are converted to ketone.
trichloropropene.
Epoxides
 more stable than the arene oxides formed from
aromatic compounds.
 susceptible to enzymatic hydration by epoxide hydrase
to form (also called 1,2-diols or 1,2-dihydroxy
compounds.

OLEFINIC EPOXIDATION
Carbamazepine (Tegretol)
 The epoxide is reasonably stable and can be
measured quantitatively in the plasma of patients
receiving the parent drug.

 Toxicity of olefinic compounds may result from their

metabolic conversion to chemically reactive epoxides.

 AFLATOXIN
 Other compounds DES, stilbene, and vinyl chloride.
 OXIDATION AT ALIPHATIC CARBON ATOMS

Oxidation at Allylic Carbon Atoms  Metabolic oxidation at the terminal or the penultimate
methyl group often is referred to as ω oxidation.
 Oxidation of the penultimate carbon atom (i.e., next-to-
the-last carbon) is called ω— I oxidation).

 Allylic hydroxylation generally does not lead to the

generation of reactive intermediates.

Oxidation at Carbon alpha to Carbonyl and Imine

 Common to benzodiazepines
 Oxidized to 3-hydroxy metabolites
 Diazepam (Valium)
 3-hydroxydiazepam (also called N-
methyloxazepam)
 Flurazepam (Dalmane)
 Nimetazepam are Anticonvulsants.

Oxidation Involving Carbon-Hetero Systems

 Hydroxylation of the alpha-carbon atom attached
directly to the heteroatom (N, O, S).
 Hydroxylation or oxidation of the heteroatom
  (N, S only, e.g.. N-hydroxylation. N-oxide
formation, sulfoxide, and sulfone formation).

OXIDATION INVOLVING CARBON-NITROGEN SYSTEMS

Three basic classes of nitrogen-containing compounds
1. Aliphatic (primary, secondary, and tertiary) and alicyclic
(secondary and tertiary) amines.
2. Aromatic and heterocyclic nitrogen compounds.
 Primary Aliphatic amines are bio-transformed by
oxidative deamination (through the carbinolamine
3. Amides pathway) or by N-oxidation.
 Susceptible to either a carbon hydroxylation or N- Enzyme: MFO
oxidation.  Endogenous primary amines (e.g. Dopamine,
 Drugs containing nitrogen norepinephrine, tryptamine, and serotonin)
 natural products Enzyme: MAO
morphine, cocaine, nicotine)  In primary aliphatic amines, such as phentermine,
 Other drugs chlorphentermine and amantadine, N-oxidation appears
 phenothiazines. antihistamines, tricyclic to be a major biotransformation pathway because a-
antidepressants, beta-adrenergic agents, carbon hydroxylation cannot occur.
phenylethylamines, benzodiazepine.
Tertiary Aliphatic and Alicyclic Amines

N- HYDROXYLATION of secondary amines generates the

corresponding N-hydroxylamine metabolites.

Primary aromatic amines are oxidized to hydroxylamines then

further oxidized to nitroso (Chlorphentermine, Aniline)
N-hydroxylation of secondary amines
 generates the corresponding N-hydroxylamine
metabolites.

Secondary and Primary Amines

 Secondary amines (either parent compounds
or metabolites) are susceptible to oxidative N-
dealkylation, oxidative deamination, and N-
oxidation reactions.
 Dealkylation of secondary amines gives rise to
the corresponding primary amine metabolite.
 ethers) is catalyzed by microsomal mixed function
oxidases.

Oxidative O-dealkylation
 It involves the oxidation of the alpha carbon.
 A
mi
de
s
 N- alkyl substituents are also dealkylated through N-

Oxidation Involving Carbon-Sulfur Systems

1. S-dealkylation
2. Desulfuration
3. and S-oxidation reactions

S-dealkylation - It also involves the alpha carbon

hydroxylation.

dealkylation.
 Hydroxylation also happens to the alpha carbon of the
amide or phosphamide.

Oxidation Involving Carbon-Oxygen Systems

DESULFURATION

 Oxidative O-dealkylation of carbon–oxygen

systems (principally
   Hydroxyl and amino groups are susceptible to
conjugation.
 Carbonyl reduction
- Alcohol
 Nitro, and azo reduction
- amino derivative
 N-oxides
- tertiary amine
 Sulfoxides
- Sulfides
 Disulfide
- -SH + -SH
Reductions of Carbonyl Containing Compounds
SULFOXIDATION  Ketones are resistant
to oxidation are

reduced
to

Oxidation of Alcohols and Aldehydes secondary alcohol.

 Aldehydes are
 Many oxidative processes (e.g., benzylic, allylic,
reduced to form
alicyclic, or aliphatic hydroxylation) generate
primary alcohol.
alcohol or carbinol metabolites as intermediate
 Enzyme : aldo-keto
products.
reductases.
 If not conjugated, these alcohol products are
further oxidized to aldehydes (if primary alcohols) Chlorpheniramine
or to

Carbinolamine (2-hydroxymedazepam)

Acetophenone
REDUCTION
 addition of hydrogen or gain of electrons. Acetohexaminde
 play an important role in the metabolism of many
compounds containing carbonyl, nitro, and azo group.
 - not recommended in diabetic patients with renal  Reduction of N-oxides to tertiary amine.
failure, because of the possible accumulation of its
active metabolite hydroxyhexamide.

 The (R) ( + ) enantiomer of the oral anticoagulant

warfarin undergoes extensive reduction of its side chain
keto group to generate the (R,S)( +) alcohol as the major
plasma metabolite.  Reduction of
sulfur-
REDUCTION OF NITRO AND AZO COMPOUNDS containing
 Leads to primary amine metabolites. functional groups, such as the disulfide and
sulfoxide.

HYDROLYSIS
• E.g
 reaction of water with substrate resulting in
- Clonazepam
breaking scissile carbon heteroatom bonds)
- Nirazepam
- Dantrolene  The reaction is frequently enzyme - mediated
- Metronidazole although serum pH may cause reaction.
Reduction of Nitro Compounds Ester hydrolysis
 Clonazepam and Nitrazepam - mediated by nonspecific esterases found in the
- are metabolized to 7-amino metabolites. liver, kidney, and intestine and
pseudocholinesterases present in plasma.
Amide hydrolysis
- mediated by
liver microsomal
amidases,
esterases and
deacylases.
 major
Azo Reductions
biotransformation pathway for drugs containing
 Proceed via a hydrazo intermediate (-NH-NH-) that
an ester functionality.
is cleaved reductively to yield the corresponding
aromatic amines.

PRONTOSIL
TARTRAZINE

AMARANTH

Miscellaneous
Reductions