Molecular Biology of

Pharmacology
Indwiani Astuti
Dept. of Pharmacology & Toxicology
Fac. Of Medicine Gadjah Mada University

Structure of DNA

The central dogma of molecular

biology
The

flow of the expression of genetic

information in cells is almost exclusively
one way: DNA RNA Protein

A gene codes for a protein

DNA

CCTGAGCCAACTATTGATGAA

transcription

mRNA

CCUGAGCCAACUAUUGAUGAA

translation

Protein

PEPTIDE

Introduction
Pharmacology:

knowledge of how drugs

interact with body constituents to produce
therapeutic effects
The elucidation of molecular mechanisms of
drug response, the development of new drugs,
& the formulation of clinical guidelines for safe
& effective use of drugs in therapy or
prevention of disease & in relief symptoms.

Pharmacological responses by molecular interaction of

drugs with cells, tissues, or other body constituents
The key word is molecular
What specific biological molecules must be present ?
How do drugs & biological molecules interact to
produce changes ?
How are these changes converted into observable
responses ?

GENETIC
POLYMORPHISMS

Pharmacokinetic
Transporters
Plasma protein binding
Metabolism

Pharmacodynamic
Receptors
Ion channels
Enzymes
Immune molecules

Efficacy & Safety

Pharmacogenomic/genetic
There

is substantial heterogenesity in
the way individuals respond to
medications, in terms of both toxicity and
treatment efficacy.
"pharmacogenetics," which initially
focused largely on genetic
polymorphisms in drug metabolizing
enzymes, and how this translates into
inherited differences in drug effects

Pharmacogenetics developed
The

first observations of genetic variation in

drug response date from the 1950s,
involving the muscle relaxant suxamethonium
chloride, and drugs metabolized by Nacetyltransferase.
One in 3500 Caucasians has less efficient
variant of the enzyme (butyrylcholinesterase)
that metabolizes suxamethonium chloride
(muscle relaxant) drugs effect is prolonged,
with slower recovery from surgical paralysis.

Variation in the N-acetyltransferase gene divides

people into slow acetylators and fast acetylators,
with very different half-lives and blood concentrations
of such important drugs as isoniazid (antituberculosis)
and procainamide (antiarrhythmic).
slow acetylator recessive
ratio fast : slow

Europe 40 : 60; USA 55 : 45

Japan 85 : 15; Oriental 10 : 90
Inuit 95 : 5

Genetic polymorphisms in drug metabolizing

enzymes

From: Evans WE, Relling MV. Pharmacogenomics: Translating functional genomics into rational
therapeutics. Science 286:487-491, 1999.

In the 1960s it became apparent that other drug

metabolising enzymes, particularly the cytochrome
P450-dependent monooxygenases, were also
polymorphic.
in the 1970s with the identification of the cytochrome
P450 CYP2D6 debrisoquine hydroxylase
polymorphism
Polymorphisms in further cytochrome P450s, as well
as in other drug metabolising enzymes, were
subsequently discovered.
With the development of recombinant DNA technology,
the molecular basis of most of these polymorphisms

 The

study of Pharmacogenomics focuses on

drug action based on a patients genotype
(DNA) and/or molecular phenotype
(DNA/RNA/protein).
Pharmacogenomics tests resultant from such
analyses will allow for the recognition of those
patients for whom a particular therapy would
be most efficacious, or for which there would
be least likely incidence of adverse events.
Thus, the major goal of pharmacogenomics is
to individualize patient therapy.

APPLICATION OF PHARMACOGENETICS TO DRUG

DISCOVERY AND USE

Most therapeutic drugs are administered orally and

need to be absorbed across the GI tract to enter the
circulation.
This process can be influenced by the presence of
drug transporters, such as the multi-drug resistance
protein, MDR1, and metabolic enzymes such as the
cytochrome P450-dependent monooxygenases, in
particular members of the cytochrome P450 CYP3A
gene family (CYP3A4, CYP3A5 and CYP2D6) (4). The
importance of the drug transporter proteins is
exemplified by the finding that marked changes in drug
absorption and pharmacokinetics are observed in mice
nulled at the Mdr1 gene locus

Pharmacogenetics and cancer

chemotherapy
polymorphism

of drug-metabolizing

enzymes
polymorphism of receptors
leads to a prolonged half-life and a
reduced elimination of cytotoxic drugs.
A consequence could be an adverse
side reaction due to an excessive drug
toxicity in non-cancer tissues.

Dehydropyrimidine dehydrogenase (DPD) is the

initial rate limiting enzyme in the catabolism of 5
fluorouracil (5FU)

located on chromosome 1p22 as a single copy and consists

of 23 exons for a total lenght of 950 kb
1 splice-site mutation, 2 deletions and 4 missense mutations.

Thiopurine S-methyltransferase (TPMT) is a

cytosolic enzyme that preferentially catalyzes the Smethylation of mercaptopurine (6MP), azothioprine
and thioguanine hematopoietic toxicity (leucopenia,
anaemia, pancytopenia)

located to human chromosome 6 and several mutant alleles

have been isolated from TPMT-deficient patients.

 Glutathione

S--transferases catalyze
the conjugation of electrophilic
compounds to cellular glutathione
absence

of GSTm 1 in children with acute

lymphoblastic leukemia is associated with a
82% remission versus only 52% in children
positive for GSTm .
Busulfan is an alkylating agent

N-acetyltransferases exist as two isoforms

respectively termed NAT1 and NAT2 and have been
initially described as the enzymes responsible for the
acetylation of isoniazid and caffeine (conversion of 1methylxanthine into 5-acetylamino 6-amino 3-methyl
uracil).

NAT2 is located on human chromosome 8q11.

Amonafide (NSC308847) fast acetylators defined

by their caffeine phenotype of NAT2 have significantly
greater leukopenia than slow acetylators, as well as
higher bone marrow toxicity at the standard dose of
300mg/m2 daily for 5 days.

Angiotensinogen: the application of pharmacogenomics to

the treatment of hypertension

The

AGT gene, in particular, has shown

the strongest and most consistent
associations (5 9) with the risk of
CHD and certain forms of hypertension.
The AGT gene, located on chromosome
1, consists of approximately 13,000
nucleotide base pairs organized into 5
exons, and 4 introns

PHARMACOGENETICS IN PSYCHIATRY

CYP2D6 genotyping in patients

receiving drugs for the treatment of
depression and psychiatric disorders.
This enzyme is responsible for the
metabolism of the majority of drugs used
in the treatment of psychiatric illness and
many of the patients suffer drug side
effects

New drugs design (for exz.)

Receptor

with intrinsic Tyrosine Kinase

Activity
Large

group of receptors for growth factors

(insulin, EGF, PDGF, hepatocyte GF etc)
Extracellular domain contains regions bind
GF, Intracellular domain contains a kinase
activity capable of phosphorylating proteins
on tyrosine residues

Herceptin (c-erb2)
Product

oncogen (EGFR)
Receptor Growth factors family
Indication: Solid tumor with Her-2 +
Breast Cancer

Tyrosine kinase sensitive

CML

expression of gene bcr-abl + (Chr.

Ph)
Imatinib (ST1571) Gleevec

BLOCK EGFR
Tyrosine kinase
inhibitors

Signal cascades are useful:

1. At each step of the cascade, the signal is amplified
2. The information that arrived at the plasma membrane in the form of a
signal is communicated to the nucleus
3. The multitude of steps enables a signal to have different effects in
different cells (because they have different target proteins)

Nanotechnology

Artificial bone, cartilage, & skin with no immune

rejection
Vitoss

(Orthovita): nanoparticels bone

growth (orthopaed)
Navavax-estrasorb (cream):
nanoparticels Skin burns
Nucrest-Silcrest : nanocrystaline Skin
burns
Nanodot (Nasa): cells repairs

Improved, direct chemotherapy and

radiotherapy
Drug

delivery

Maximizing

bioavailability both over a period

of time and at specific places in the body
Deliver drug directly to the site without
interacting with the rest of the body
Smart drugs

Nanotechnology for Drug Delivery

Molecule

encapsulated within nanoscale

cavities inside polymer : time-released
drugs
Grind solid drugs into fine powders : to
increase surface area and reactivity & to
increase solubility
Encapsulate polar drug in a nonpolar
coating : easily pass through the cell
membrane

Coat DNA with cholesterol to easily pass through the

oily cell membrane
Liposome structures to deliver soluble protein
(cytokine) such as interferon to cancer cells
Magnetic nanoparticles : local bioavailability control by
external magnetic field
Triggered response : inactive drug moleculewakes
up on encountering a particular signal
Antacid enclosed in a coating of polymer that dissolves
only in highly acidic conditions