DRUG

METABOLISM
(PHASE II)
Dr. Mohammad Javed Naim
Assistant professor
Pharmaceutical Chemistry-I
Semester I
Lecture 2
Date: 16 Oct 2023
Outline
◦ Phase II metabolism
◦ Drug metabolizing enzymes
◦ Factors affecting drug metabolism
Objectives
◦ Our main aim is to study Phase II metabolic reactions in detail with its various metabolizing enzymes and factors
affecting metabolism so that we can easily understand the process of metabolism and what kind of drug molecule
are getting metabolized by which pathway.
 Phase II metabolism
❖The activated xenobiotic metabolites are conjugated with charged species such as glutathione (GSH),
sulfate, glycine, or glucuronic acid.
❖Sites on drugs where conjugation reactions occur include carboxy (-COOH), hydroxy (-OH), amino (NH2),
and thiol (-SH) groups.
❖Products of conjugation reactions have increased molecular weight and tend to be less active than their
substrates, unlike Phase I reactions which often produce active metabolites.
❖The addition of large anionic groups (such as GSH) detoxifies reactive electrophiles and produces more
polar metabolites that cannot diffuse across membranes, and may, therefore, be actively transported.
❖These reactions are catalysed by a large group of broad-specificity transferases, which in combination can
metabolise almost any hydrophobic compound that contains nucleophilic or electrophilic groups.
 Reactions

❖Methylation

❖Sulphation/ Sulfate conjugation

❖Acetylation

❖Glucuronidation

❖Glutathione conjugation

❖Glycine conjugation
 Mechanism Involved enzyme Co-factor Location
Methylation Methyltransferase S-adenosyl-L-methionine Liver, kidney, lung, CNS
3'-phosphoadenosine-5'-
Sulphation Sulfotransferases Liver, kidney, intestine
phosphosulfate

•N-acetyltransferases
Acetylation •bile acid-CoA:amino acid N-
acyltransferases
Liver, lung, spleen, gastric
Acetyl coenzyme A
mucosa, RBC, lymphocytes

Liver, kidney, intestine, lung,

Glucuronidation UDP-glucuronosyltransferases UDP-glucuronic acid
skin, prostate, brain

Glutathione conjugation Glutathione S-transferases Glutathione Liver, kidney

1.Two step process:XM-

ligase (forms a xenobiotic acyl-
CoA)
Glycine conjugation Glycine lLver, kidney
2.Glycine N-
acyltransferase (forms the
glycine conjugate)
Methylation

❖Most of the endogenous amines undergoes methylation reaction with the help of methyltransferase

enzyme. E.g. COMT (Catechol Ortho Methyl transferase)

 Sulfation/Sulfate conjugation

❖Drugs having hydroxyl group, Phenol and aromatic amines undergoes sulfate conjugation in the presence

of enzyme sulfotransferase.

❖Most of the steroidal drugs undergoes metabolism by sulfate conjugation.

 Acetylation

❖Acetylation involves acetylation with acetyl coenzyme A in the presence of enzyme acetyl transferase

which is present in liver, spleen and RBCs..

❖Drugs containing amino or hydrazine functional group undergoes acetylation.

Glucuronidation
❖ It involves metabolite conjugation with activated form of glucuronic acid.

❖ Molecules associated with alcoholic hydroxyl, phenolic hydroxyl and carboxylic group undergoes

metabolism by this pathway..

Glycine conjugation
❖ Most important route for conjugation of drug for elimination.

❖ Glycine is the most important amino acid which forms water soluble ionic conjugates with aliphatic,

aromatic and heterocyclic carboxylic acids.

Glutathione conjugation
❖ Important in elimination of polycyclic phenols and halides.

❖ It is catalysed by enzyme glutathione transferase.

Inhibition of drug metabolism
❖One drug can inhibit the metabolism of another drug.

❖Increase in circulating levels of slowly metabolized drug.

❖Prolongation or Potentiation of its effects.

❖Consequences:

❖Precipitation of toxicity

❖Can be therapeutically beneficial. E.g. Aversion of Alcohol with Disulfiram

◦ Comparison between Phase I and Phase II metabolism
 ◦ Phase I ◦ Phase II

❖Oxidation (via cytochrome P450), reduction,
and hydrolysis reactions.
❖Phase I reactions convert a parent drug to more
polar (water soluble) active metabolites by
unmasking or inserting a polar functional group
(-OH, -SH, -NH2).
❖Geriatric patients have decreased phase I
metabolism.
▪ Drugs metabolized via phase I reactions have
longer half-lives
▪ Geriatric patients metabolism drugs by phase
II reactions
❖ Glucuronidation, acetylation, and sulfation reactions.
❖Conjugation reactions" that increase water solubility of
drug with a polar moiety
❖Glucuronate, acetate, and sulfate, respectively
❖Phase II reactions convert a parent drug to more polar
(water soluble) inactive metabolites by conjugation of
subgroups to -OH, -SH, -NH2 functional groups on drug.
❖Drugs metabolized via phase II reactions are renally
excreted
❖Patients deficient in acetylation capacity (slow
acetylators) may have prolonged or toxic responses to
normal doses of certain drugs because of decreased rates
of metabolism
◦ Drug metabolizing enzymes
Role of cytochrome P450 Monooxygenases in
Oxidative Biotransformation's
❖General Stoichiometry that describes the oxidation of many xenobiotics (R-H) to their corresponding

oxidised metabolites (R-OH) is given by the following chemical reaction:

❖The enzyme system carrying out this biotransformation are referred to as mixed function oxidases or

monooxygenases.

❖The reaction requires both molecular oxygen and NADPH.

❖During this oxidative process, one atom of molecular oxygen is introduced into the substrate (R-H) to give

R-OH and the other atom is incorporated to form water molecule.

❖Cytochrome (CYP) enzymes are responsible for transferring the Oxygen atom to the substrate.
❖These cytochrome (CYP) enzymes are haeme proteins.

❖The haeme portion is an iron containing porphyrin called protoporphyrin IX and the protein portion is

called apoprotein.

❖ Cytochrome (CYP) is found in high concentration in live (major organ involved in the metabolism of

xenobiotics).

❖The presence of this enzyme in many other tissues (Lung, kidney, intestine, skin, placenta, adrenal cortex)

shows that these tissues have drug oxidising capability too.

❖ The CYP monooxygenase are located in the endoplasmic reticulum. Many of the cytochrome enzymes are

responsible for the biosynthesis of steroidal hormones and metabolism of certain vitamins.
CYP450 Oxidation reduction cycle
◦ Factors affecting drug metabolism
A large no. of physical, chemical and biological factors affects the metabolism of drugs which are as

mentioned below

1. Physico-chemical properties of drug:

Size, shape, acidity, basicity, solubility, lipophilicity and pKa value of the drug.

2. Chemical factors:

Various chemicals affects drug metabolism-Presence of enzyme inducers and inhibitors alters the action of

drug metabolizing enzymes.

Enzyme induction (Chemical inducers): These are the chemicals which in creases the activity of the enzymes.

E.g. 3-methylchloranthene and cigarette smoking increases the metabolism of certain drugs.

Alcohol increases metabolism of coumarins and phenytoin's and barbiturates increases metabolism of oral

contraceptive drugs and cortisol.

Rifampicin and isoniazid stimulates their own metabolism (self induction of metabolism).

Enzyme inhibitors: Chemical which decreases the activity of enzymes responsible for metabolism. Inhibition

of enzyme may be direct, indirect, comparative or non-comparative.

MAO inhibitors decreases the metabolism of barbiturates.

Coumarins decreases the biotransformation of phenytoin.

3. Environmental factors: Temperature, Pressure, Atmospheric humidity.

4. Biological/ Physiological factors: Age of patient, diet, gender, altered physiological state like

pregnancy, disease or hormonal imbalance.

5. Pharmacodynamic factors: Dose, frequency, route, tissue distribution and protein binding.

6. Stereochemical factors: Depend on the stereochemistry of the drug molecule and stereoisomerism as

well.

7. Genetic factors: Biotransformation and conjugation exhibit species variation leading to extended or

excessive therapeutic effect or overdose.

Lipid water partition coefficient

❖The lipid/water partition coefficient denotes the ratio of the concentration of a drug in two immiscible or

slightly miscible phases.

❖The distribution law is exact only for ideal solutions under the following conditions:

✓ When the two liquid phases are completely immiscible;

✓ When the solute neither associates nor dissociates in either phase;

✓ When the solute concentration is relatively low;

✓ When the solute is only slightly soluble in either phase.

References

❖Ionescu, C., & Caira, M. R. (Eds.). (2005). Drug metabolism: current concepts.

❖Khan, M. F., & Philip, A. (Eds.). (2018). Fundamentals of medicinal chemistry and drug metabolism (Vol.

1). Bentham Science Publishers.

❖Abraham, D. J. (2006). Burger's medicinal chemistry and drug discovery.