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Re: Pharmacogenomics

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Respected members,

Pharmacogenetics involves the study of single gene mutations and their effect on

drug response. The term pharmacogenomics is much broader and it involves

surveying the entire genome to assess several determinants of drug responses.

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Genotype is the genetic constitution of an organism (specific set of alleles are

inherited at a locus)

Phenotype is the observable biochemical or physiological characteristics of the

expression of the gene.

There is association between the presence of a certain mutations (genotype) and

the resulting physical trait, personality, abnormality or pattern of

abnormalities (phenotype).

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Respected members,

Some basic concepts are,

- Three billion base pairs in human genome

- These bases in the DNA form the genotype of an individual.

- Genotype is responsible for the expression of phenotype.

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If mutation of gene occur i.e. permanent, heritable change in the genetic

sequence subsequently phenotype may also change, but it is not compulsory that

every mutation will be expressed phenotypically. Mutations could be due to:

- Single base substitution

- Insertions or deletions

- Chromosomal mutations

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Respected members,

Variations in the DNA sequence that is present at an allele and is with

frequency of & #8805;1% in a human population. It can be SNPs(Single nucleotide

polymorphism) or INDELS (Insertion, deletion)

SNP occur when a single nucleotide (A,T,C,or G) in the genome

sequence is altered, e.g., AAGGCTAA to ATGGCTAA

There is approximately one SNP per 1000-2000 base pairs. SNPs exist every 100 to

300 bases along the 3-billion-base human genome.

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SNPs Comprise 90% of all human genetic variation. They are found in both

coding (i.e., gene) and noncoding regions of the genome.

Most of the SNPs are situated in the intron(noncoding) part of the DNA.

So usually they have no effect on cell function, but some could predispose

people to disease or influence their response to a drug.

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Respected members,

There are 2 main mechanisms for altered drug response.

1. Pharmacokinetic

2. Pharmacodynamic

Pharmacokinetic variation is at level the transport and metabolism.

a.Transport--in the past 17 years have pharmacologists became

fully aware of the impact of transmembrane drug transporters

on the uptake of drugs from the gut into the human

body and from the circulation into the target tissues and

into the metabolizing and eliminating organs. This carriermediated

transmembrane transport becomes particularly

important in molecules with larger molecular diameters

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Respected members,

Some substrates for P-gp are

& #61591; Anticancer drugs-Actinomycin D, Vincristine.

& #61591; Cardiac drugs- Digoxin, Quinidine.

& #61591; HIV Protease inhibitors- Ritonavir, Indinavir.

& #61591; Immunosuppresants- Cyclosporine, Tacrolimus

& #61591; Antibiotics- Erytromycin, Levofloxacin

Lipid Lowering agents- Lovastatin, Atorvastatin

Regards,

Dr. Amruta Dawari

JR1 Dept. of Pharmacology,

GMCH, Nagpur.

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Respected members,

Some of the best studied metabolizing enzymes in relation to altered drug

responce are

- & #151;Cytochrome P450

& #151;-N-acetyl transferase isoenzyme

& #151;-UDP glucuronoyl transferase

& #151;-Methyl transferase

Regards,

Dr. Amruta dawari

JR1,Dept of Pharmacology

GMCH,Nagpur

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Respected members,

CYP 450 are most important & major metabolizing enzymes. There is

large Intra and Interspecies variability. There are

- 12 families of CYP 450 in mammels

- 30 families of CYP 450 in humans

The code is same for genes which codes for protein

CYP 1 – CYP 4 are mostly used for metabolism of various drugs

Same drug may be metabolised by two Iso-enzymes e.g.

& #61607; Imipramine Hydroxylation occurs by CYP 2D6

& #61607; Imipramine Demethylation occurs by CYP 1A2, CYP 2C19

Dr. Amruta Dawari

JR1 Dept. of Pharmacology,

GMCH, Nagpur

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Respected members,

CYP 3A4 metabolize 50% of all drugs & CYP 2D6 metabolize 20% of all drugs.

Unlike other P450 (e.g. CYP 2D6) there is no evidence of deletion/null allele

for CYP 3A4. Variance in coding region of CYP 3A4 occur at allele frequency of

less than 5% & appear as heterozygous with wild type allele. These coding

varicosity may contribute to, but are not likely to be major cause of

interindividual difference of CYP3A4 dependent clearance. There is need to

further explore CYP 3A4 polymorphism and it's functional impact.

Dr. Amruta Dawari

JR1 Dept. of Pharmacology,

GMCH, Nagpur

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Respected members,

Some of the mutant alleles of CYP 2D6*1 are:

*1XN,*2XN which show increased enzymatic activity.

*3,*4,*5,*6 which show absence of enzyme.

*9,*10,*17 show decreased enzyme activity.

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Substrate for CYP 2D6 are, clonidine, codeine, promethazine, propranolol,

clozappine, fluoxetine, haloperidol, amitryptiline.

Mutant alleles of CYP 2C19*1 are *2, *3, *4, *5, *6, *7, *8 which show absence

of enzye & substrates for this enzyme are

proguanil, imipramine, ritonavir, nelfinavir, cyclophosphamide

Mutant alleles of CYP 2C9*1 are *2, *3, *4, *5, *6 which sow reduced activity of

enzyme & substrate for this enzyme are warfarin, losartan, phenytoin,

tolbutamide.

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NAT2 (n-acetyl transferase 2)show considerable heterogeneity worldwide

Eg. Slow acetylator phenotype frequency is

50% in American whites and Blacks

60-70% in North Europeans

5-10% in Southeast Asians

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1.TPMT: 6-Mercaptopurine, 6-thioguanine, azathioprine

2.Dihydropyrimidine dehydrogenase (DPD) 5-Fluorouracil, capecitabine

3.Uridin diphospho-glucuronic acid

transferase type (1A1)UGT1A1: Bilirubin, irinotecan

4.Vitamin K epoxide reductase (VKORC1): Warfarin, acenocoumarol, phenprocoumon

5. NAT (NAT 2) :INH, Hydralazine,Sulfonamides,Procainamide

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Respected members,

Here are some examoles of pharmacodynamic variations of drug response.

There are genetic polymorphisms in the gene coding for the

beta1 adrenergic receptor (ADRB1) and in that coding for the

beta2 receptor (ADRB2). In ADRB1, a Ser49Gly variant

may be associated with enhanced agonist-induced downregulation,

whereas a Gly389Arg variant was found to bring

about a fourfold higher agonist stimulated signal transduction

to the Gs protein compared with the Gly allele.

Dr. Amruta Dawari

JR1 Dept. of Pharmacology,

GMCH, Nagpur

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Respected members,

Severe (and in some cases fatal) allergic reactions

to the HIV nucleoside analogue drug abacavir have been

explained by the HLA-B*5701 allele, and a HLA-B*5701,

HLA-DR7, and HLA-DQ3 haplotype even had positive and

negative predictive values of 100 and 97%, respectively

, strongly advocating routine testing before abacavir

prescription. Such a testing may not only be medically

needed, but it may also be cost-effective.

Amruta Dawari

JR1 Dept. of Pharmacology,

GMCH, Nagpur

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Respected members,

Identification of pharmacogenomic markers

1)Linkage study: Genotyping families with micro satellite-markers to correlate

inheritance of particular chromosomal region with inheritance of disease in

family members.

Eg. Malignant hyperthermia

But it is impractical.

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Respected members,

2)Association studies

Correlate the presence of chromosomal region and trait in unrelated individuals

of a population.

Eg. Apo E4 is strongly associated with Alzheimer's disease

Dr. Amruta Dawari

JR1 Dept. of Pharmacology,

GMCH, Nagpur

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Respected members,

3)DNA micro arrays (DNA chips)is anevolving technology, by this method it

is possible to examine patients for presence of specific SNP's quickly and

affordably.

Microarray solid-phase bound DNA molecules to simultaneously genotype large

numbers of SNPs

(up to more than a million) in a single sample. Used in the genome-wide

association studies.

Dr. Amruta Dawari

JR1 Dept. of Pharmacology,

GMCH, Nagpur

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Respected members,

Genomics versus epigenomics

We (scientists and physicians engaged in pharmacogenetics

and genomics) have learned and repeatedly taught the

following: analyzing inherited genetic variation has the big

advantage that analysing the genome of one cell of an

organism provides reliable information on all the other cells

independent of age, tissue localization or environmental

factors. However, there are somatic cell mutations as well as

tissue-specific epigenetic effects, such as DNA methylation,

histone modification or micro-RNA expression, that significantly

and constantly can change the pattern of gene

expression of the cell. Such changes can significantly modify

drug efficacy or initiate adverse effects and, therefore, they

have to be taken into account in future clinical pharmacology.

Dr. Amruta Dawari

JR1 Dept. of Pharmacology,

GMCH, Nagpur

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Respected members,

The concept of personalized medicine evolves from pharmacogenomics.

What is personalized medicine?

Developing drugs on the basis of individual genetic differences

How does it work?

-Tailoring therapies to genetically similar subpopulations results in improved

efficacy and less toxicity.

Regards,

Dr. Amruta Dawari

JR 1, Dept of pharmacology,

GMCH, Nagpur.

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Respected members,

In personalized medicine with the help of genetic information we can give

drug to only those patient who are likely to respond to drug & avoid giving drug

to non responders & patients likel to have toxicity of that drug.

Regards,

Dr. Amruta Dawari

JR 1, Dept of pharmacology,

GMCH, Nagpur.

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Respected members,

Chalanges to be met by pharmacogenomics are,

1. Cost of gene tasting e.g. a simple test for CYP may cost you 200-1000 $

2.Polygenic nature of diseases

3.Tests for genetic polymorphism are not standardized. Most of them lack

specificity.

4.Physicians lack awareness.

5.Complexity during prescription

6.There is no regulation regarding genetic testing

7.Patients have fear regarding genetic testing

Regards,

Dr. Amruta Dawari

JR 1, Dept of pharmacology,

GMCH, Nagpur.

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