PHARMACOL

OGY PHL 313

Chapter 4
Biotransformation (Metabolism)
 Excretion can terminate the activity of some, but

Why the body not all, drugs

• Renal excretion is good for polar molecules or
needs to small molecular volumes
• Organic drugs tend to be lipophilic and non
metabolize ionizable (or minimally ionized) at physiological
pH
drugs  readily reabsorbed from glomerular filtrate

? • Termination of these drugs is expedited by

transformation to more water-soluble molecules
 In the body parts
• Primarily in the liver
• Also in the:
• GI (Gastro-intestinal)
• Stomach and upper intestines, endogenous process
tract

Locations of • Lung
• Lower intestine assisted by micro-organisms

the metabolism • kidney

• Brain
• Skin
In the cellular Level
Metabolism is a cellular process, occurring in the smooth
ER/microsomes or in mitochondria, lysosomes, the cytosol or the
nucleus
 Does metabolism always mean inactivation?
No, there are three possibilities
1. Often, metabolites are less efficacious than
their parent compounds, or they may have no
activity
Fate of the drugs 2. Other metabolites may have enhanced activity
(metabolism may be exploited with pro-drugs,
use physiological process to activate drugs), i.e.
metabolism = activation
3. Some metabolites may have increased and
excessive toxicities
 Occurs through two phases (steps)
Phase I reactions
• Introduce enhanced polarity / unmask a functional group

How the • Generally add / reveal: -OH, -SH, -NH2

• Most often inactive metabolites

metabolism • If enough polarity is added, may be ready for excretion

occur in Phase II reactions

• Conjugations are made between phase I modified, or parent drugs

the body • Several conjugates are possible, including glucuronidation,

sulfation, acetic acid conjugation and more..

Important Note: Phase II may precede Phase I reactions (They do not to be

organized)
Phases of drug metabolism
 Phase I metabolism means Introduction of a polar functional
group (like -OH, -NH2, -SH), or modification of existing functional
group in a drug molecule such that it becomes more polar than
before.

Phase I Types of phase I metabolism reactions include:

Functionalizatio 1. Oxidation 2. Reduction 3. Hydrolysis 4. Isomerization

n reactions The main function is to make the compounds more polar and
hydrophilic to be easily excreted, and to prepare compounds for
phase II metabolism
Results typically is the loss of pharmacological activity of the
parent drug, but sometimes may be equally or more active than
parent drug, or it may be converted to a toxic material
 The smooth endoplasmic reticulum (SER) is the home of
a lot of phase I metabolic reactions
Can form vesicles called microsomes when cell is
homogenized, and they contain enzymes still capable of
metabolism, with special enzymes classes:

Phase I • Mixed function oxidases (MFO)

• Monooxygenases
Location in Enzyme activity requires a reducing agent (NADPH) and
molecular oxygen (O2).
the cellular NADPH: Nicotinamide adenine dinucleotide phosphate,
level abbreviated NADPH is a coenzyme used in anabolic
reactions, such as lipid and nucleic acid synthesis, which
require NADPH as a reducing agent to initiate the
reaction.
 1. NADPH-cytochrome P450 oxidoreductase (POR):
It is a membrane-bound enzyme required for
electron transfer from NADPH to cytochrome P450 in
the endoplasmic reticulum

Phase I 2. Cytochrome P450, a hemoprotein = P450 or CYP

Key enzymes:
They are a superfamily of enzymes containing heme
microsomal as a cofactor that function as monooxygenases,
these proteins oxidize steroids, fatty acids, and
enzymes xenobiotics, and are important for the clearance of
various compounds, as well as for hormone synthesis
and breakdown
Phase I
Nomenclature of CYP450
Enzymes
 A “super-family” of enzymes with a broad
substrate selectivity, based on shared homology
of amino acid sequence
 Categorized into 17 families, over 50 isoforms
identified (sequences > 40% identical)
 CYP1, CYP2, CYP3, or CYP4
 Subfamilies exhibit >55% sequence similarity
 identified by a letter, CYP1A, CYP2D
 May have different isoforms, another
Arabic number, CYP2D6, CYP3A4
 Capital letters human and small
letters animals

 A gene is said to be polymorphic if more

than one allele occupies that gene’s locus
within a population
Phase I
Isoforms of
Enzyme
CYP450
s
There are numerous isoforms of P450s
Most important / predominant forms
are:
• 1A2 (8.9%)
• 2A6 (3.4%)
• 2B6 (7.2%)
• 2C9 (12.8%)
• 2D6 (20%)
• 2E1 (3%)
• 3A4 (30%) Responsible For >50% Of
hepatic Xenotoxic metabolism
 Phase II Metabolism means conjugation reactions
occur in which a polar molecule is linked covalently
to a suitable functional group on a drug or one of
its Phase I metabolites so the drug or the
Phase II metabolite becomes easily excreted because the
physicochemical properties of that drug have
Conjugatio changed due to conjugation

n reactions Results will vary depend on the drug and the

metabolites, they may becomes easily excreted, and
loose their activity or they still active or even change
by increasing or decreasing the activity. Highly polar,
rapidly excreted in urine and feces
 Enzymes (transferases) located predominantly in
cytosol; products are usually inactive (exception is
Phase II morphine 6-glucuronide)
Enzyme activity Phase II reaction require UDP-GA as
Location in a cofactor

the cellular Note, Phase II reactions are generally faster than

level Phase I
 1. Glucuronic acid conjugation
2. Sulfate conjugation
Phase II 3. Amino acid Glycine conjugation
Conjugatio 4. Glutathione conjugation

n reactions 5. Acetylation
6. Methylation
 1- Glucuronic acid conjugation: For alcohols, phenols, carboxylic acids.
Enzyme: glucuronyl transferase
• Most widespread, important of the conjugation reactions
• Cofactor UDP–Glucuronic acid is in high abundance
• Closely related to glycogen synthesis
Phase II • Found in all tissues of the body
• Other sugars, glucose, xylose or ribose can be conjugated

Key 2- Sulfate conjugation: For alcohols, phenols, amines. Enzyme:

microsomal sulfotransferases
• Major conjugation pathway for phenols, also alcohols and amines

enzymes • Compounds that can be glucuronidated can also be sulfated, there is

competition between the two pathways.
• In general, sulfate conjugation predominates at low substrate
concentration and glucuronide conjugation predominates at high
substrate concentration
P450 enzyme
induction and
inhibition
Beyond being metabolized by a CYP enzyme, many drugs can affect CYP expression or
function, altering their own metabolism, or that of other drugs
• INDUCTION
• Some drugs can increase the rate of CYP synthesis/decrease the rate of
degradation
• Enhance their own metabolism, and that of other drugs being co-
administered
• Can do so by increasing transcription, translation, or adding heme
groups
• INHIBITION
• Drug can also inhibit function of CYPs, leading to decrease metabolism of
themselves, and co-administered drugs
• May be irreversible/covalent binding  suicide inhibitors
P450 enzyme
induction
and
inhibition
P450 enzyme
induction
and
inhibition
Toxic
Byproducts

Not all
metabolism is
for clearance
 Individual differences:
1. Just as variant as ED50 and LD50 doses, so are
Clinical metabolic rates

Relevanc 2.
3.
Genetics
Drug-drug interactions
e
 Often autosomal recessive traits
• Phase I polymorphisms
• Many CYP enzymes
• Can be Poor Metabolizers (PM)
• Can be ultra rapid metabolizer (URM)
• Can be extensive metabolizer (EM) – middle
• What is effect on drug efficacy, activation,

Genetics toxicity with each?

• Phase II polymorphisms
• Can be slower enzyme reactions (i.e. slow acetylator
phenotype)
• Can be deficient in enzyme
• The role of testing in drug efficacy and safety
• May be important in determining extreme cases of
toxicity, activation, or drug efficacy
• TPMT with 6-MP
Variations
in phase I
metabolism
 • Drugs can act as substrates, inducers, or inhibitors
• Effect of induction/inhibition on not only self-
metabolism, but cross-metabolism is clear
• May affect co-administration, or even
delayed administration of ‘other’ drugs
• Also may have cross-over/competition of co-

Drug-drug administered compounds

• Not likely to affect later drug

interactions administrations
• Overall, may see need for doses
changed of 1, or both,
drugs
• Sometimes metabolism is inhibited, may also be
enhanced (see next slides)
• If discontinue one of co-administered drug, may need
to dose modify ‘other’ to avoid rapid changes in *its*
metabolism
Co-morbid
conditions affecting
metabolism
• Hepatic disease *may* modify
metabolism (depends on
pathology)
• Cirrhosis, hepatitis, even
limitations in blood flow can
affect metabolism
• May also see alterations in
metabolism from pulmonary
insufficiency, endocrine
dysfunction, diabetes, and
inflammation