Angela B.

Telesforo

 OBJECTIVES:
1. To discuss the biotransformation of xenobiotics.
2. To explain the role of enzymes in the biotransformation of
xenobiotics.
3. To differentiate Phase 1 from Phase 2 reactions.
4. To compare the different reactions involved in Phase 1
and Phase 2 reactions.

What is Biotransformation?
It is the conversion of chemicals to a more
water soluble compounds.
Xenobiotic a chemical compound ( drug, pesticide,
carcinogen ) that is foreign to a living
organism.
Endogenous chemical growing or originating from
within.

Substrate substance to be catalyzed

Substrate

enzyme
co-enzyme

transformed product

Ex.
ethyl alcohol
(CH3CH2OH)

alcohol
acetaldehyde
dehydrogenase (CH3CHO)

 Enzymes play a vital role in biotransformation

Transformation of Xenobiotics either be beneficial

or harmful
Depending on the dose and circumstances
Phase 1

- addition of a functional group.

Phase 2

- conjugation of the modified xenobiotic

with another substance.

Conjugated Products
larger molecule than substrate

Generally polar in nature ( water soluble )

Have poor ability to cross cell membranes

Phase 1 Reactions
HYDROLYSIS

- reaction with the addition of water ( OH + H)

( esters, amines, hydrazines, carbamates )

ex.
procaine

p-aminobenzoic acid +
diethylaminoethanol

Enzymes involved in
Hydrolysis
Carboxylesterases ( serum & tissues )
- hydrolyze endogenous lipid compounds
- generate pharmacologically active metabolites
Cholinesterases
- limit the toxicity of organophosphates
Epoxide hydrolase
- detoxify electrophilic epoxides ( cause cellular
toxicity and genetic mutations )

 REDUCTION

- substrate gains electrons

- occur with xenobiotics in which oxygen content
is low
- reduction reactions frequently result in
activation of a xenobiotic than detoxification

Ex.
Azo reduction nitrogen-nitrogen double bonds
Nitro reduction NO2
catalyzed by:
* CYP450
* NADPH-quinone oxidoreductase

ex.
nitrobenzene + H2

aniline + O2

OXIDATION

- reactions in which substrate loses electrons

* oxygenation
*dehydrogenation
*electron transfer

Enzymes involved in
Oxidation
Alcohol dehydrogenase
primary alcohols
secondary alcohols
Aldehyde dehydrogenase
aldehydes

aldehydes
ketones

carboxylic acids
( NAD cofactor )

Monoamine oxidase ( MAO )

oxidative deamination of primary, secondary, and
tertiary amines, including serotonin and
some xenobiotics.

Prostaglandin H synthetase
( cyclooxygenase )
arachidonic acid
prostaglandins

Cytochrome P450 ( CYP )

- found in hepatic ER microsomes
- heme containing
- classified into subfamilies based on amino acid
sequence identity
- named in a species-specific manner

 Factors that contribute to Decreased CYP enzyme

activity
1. A genetic mutation gives rise to the poor and
intermediate metabolizer genotypes
2. Exposure to an environmental factor ( infectious

disease or an inflammatory process ) - suppresses

CYP enzyme expression
3. Exposure to a xenobiotic - inhibits or inactivates a

preexisting CYP enzyme

 By inhibiting cytochrome P450, one drug can

impair the biotransformation of another leading
to an exaggerated pharmacologic or toxicologic
response to the second drug

 Factors that contribute to Increased enzyme activity

1.

Gene duplication leading to over-expression of a

CYP enzyme

2. Exposure to drugs and other xenobiotics that

induce the synthesis of cytochrome P450

3. Stimulation of preexisting enzyme by a xenobiotic

Induction of cytochrome P450 by xenobiotics increases

CYP enzyme activity

By inducing cytochrome P450, one drug can

stimulate the metabolism of a second drug and
thereby decrease or ameliorate its therapeutic

effect.

 Environmental Factors known to affect CYP levels

Medications
Foods
Social habits ( alcohol consumption, cigarette

smoking )
Disease status ( diabetes, inflammation, viral &
bacterial infection, hyperthyroidism,
hypothyroidism )

It is possible that two or more CYP enzymes can

contribute to the metabolism of a single compound.

 Information on which human CYP enzyme metabolizes

a drug can help predict or explain drug interactions
Inducers of cytochrome P450 increase the rate of

xenobiotic biotransformation

 P450 induction lowers blood levels, which compromises

the therapeutic goal of drug therapy but does not cause
an exaggerated response to the drug
P450 induction can cause pharmacokinetic tolerance
whereby larger drug doses must be administered to
achieve therapeutic blood levels due to increased drug

biotransformation

Phase II Reactions
CONJUGATION
Conjugations result in a large increase in xenobiotic

hydrop0hilicity greatly facilitates excretion of foreign

chemicals. ( except methylation & acetylation)
Most conjugation enzymes are mainly located in the

cytosol.

 Glucuronidation
Requires the cosubstrate uridine diphosphate-

glucuronic acid ( UDP-glucuronic acid )

Reaction is catalyzed by UDPglucuronosyltransferases ( UGTs )
Endogenous substrates include bilirubin, steroid
hormones, and thyroid hormones
Conjugates of are polar, water-soluble metabolites
Excreted from the body in bile or urine

Cofactor availability can limit the rate of

glucuronidation of drugs that are administered in

high doses and are conjugated extensively, such as
aspirin and acetaminophen

 Sulfonation ( sulfate conjugation )

Catalyzed by sulfotransferases which produces a

highly water-soluble sulfuric acid ester

The cosubstrate for the reaction is
3-phosphoadenosine-5-phosphosulfate (PAPS)
which is synthesized from inorganic sulfate
Involves the transfer of sulfonate from PAPS to the
xenobiotic
Conjugates are excreted mainly in urine

 Sulfonation is an effective means of decreasing

the pharmacologic and toxicologic activity

of xenobiotics

 Methylation
Minor pathway of biotransformation
Decreases the water solubility of xenobiotics
Masks functional groups that might otherwise be

conjugated by other enzymes

The cosubstrate for methylation is
S-adenosylmethionine ( SAM )
Methylation can also lead to increased toxicity
O-Methylation, N-Methylation, S-Methylation

 Acetylation
N-acetylation is a major route of biotransformation

for xenobiotics
aromatic amine
hydrazine

aromatic amide
hydrazide

N-acetylation of certain xenobiotics, such as

isoniazid, facilitates their urinary excretion

N-acetylation is catalyzed by cytosolic

N-acetyltransferases ( NAT ) requiring the

cosubstrate acetyl-coenzyme A ( acetyl-CoA )
NAT1 and NAT2 ( acetyltransferases in humans )
Slow NAT2 acetylators are predisposed to drug

toxicities

Drug toxicities
Excessive hypotension from hydralazine
Peripheral neuropathy from isoniazid and dapsone
Systemic lupus erythematosus from hydralazine and

procainamide
Toxic effects of coadministration of anticonvulsant
phenytoin with isoniazid

 Amino Acid Conjugation

2 pathways

conjugation of xenobiotics containing:

Carboxylic acid group with the amino group of

amino acids glycine, glutamine and taurine

Aromatic hydroxylamine with the carboxylic

acid group of amino acids serine and proline

Amino acid conjugates of xenobiotics are eliminated

primarily in urine
Conjugation of hydroxylamines with amino acids is

catalyzed by cytosolic aminoacyl-tRNA synthetases

and requires ATP

 Glutathione conjugation
Tripeptide glutathione comprises of glycine,

cysteine, and glutamic acid

Catalyzed by a family of glutathione
S-transferases that are present in most tissues
Types of conjugation reactions

Displacement reactions glutathione displaces an

electron-withdrawing group
2. Addition reactions - glutathione is added to an
activated double bond or strained ring system
1.

 Glutathione conjugates formed in the liver can be

effluxed into bile and blood, and they can be

converted to mercapturic acids in the kidney and
excreted in urine
Conjugation with glutathione represents an important

detoxication reaction :
- because electrophiles are potentially toxic
species that can bind to critical nucleophiles
( proteins & nucleic acids ) causing cellular
damage and genetic mutations.

Glutathione is a cofactor for glutathione peroxidase,

important in protecting cells against lipid and

hemoglobin peroxidation
Conjugation with glutathione enhances the toxicity of

a xenobiotic by:
1. releasing a toxic metabolite
2. Being inherently toxic
3. Being degraded to a toxic metabolite