ENUGU STATE UNIVERSITY OF SCIENCE

AND TECHNOLOGY
COLLEGE OF MEDICINE
FACULTY OF BASIC MEDICAL SCIENCES
DEPARTMENT OF MEDICAL
BIOCHEMISTRY
XENOBIOTIC BIOTRANSFORMATION
By

Martins Obinna Ogugofor

 XENOBIOTIC BIOTRANSFORMATION

• The term ‘‘Xenobiotic’’ is coined from the two words:

• Xeno: foreign/ different origin
• Biotic: relating or resulting from organisms
• Thus, xenobiotics are referred to as compounds found in an
organism but are not naturally synthesized by the organism or
naturally present in the environment.
• Examples of xenobiotics include pesticides, pharmaceutical
compounds, personal care products, illicit drugs, industrial
products, and nuclear waste.
Examples of Xenobiotics
 Exposure To Xenobiotics
• Xenobiotic exposure normally occurs via the following routes:
• Inhalation (Lungs)

• Ingestion (Gut)

• Adsorption (Skin)

• Injection (Muscle/circulation)
 ORAL ROUTE
• One of the most important absorption site.
• Important for food/water contaminants and drugs.
• Absorption of xenobiotics can take place along entire GI tract.
• To enter the body via the GIT, chemicals must pass through the GIT lining
and capillary membranes before entering the blood.
• The greatest absorption of xenobiotics takes place in the small intestine
especially alkaline toxicants due to its large surface area.
• The rate of absorption increases with the residency time. The residency time
of a xenobiotic in the small intestine is dependent on intestinal motility.
 INHALATION AS A ROUT OF
EXPOSURE TO XENOBIOTICS
• Xenobiotics such as gases, vapors of volatile liquids and aerosols are
absorbed by this route.

• The absorption of gas depends on its solubility in blood.

• The physical form and particle size of the xenobiotic determines penetration.

• Particular very small particles are able to reach alveoles and can enter
bloodstream.
 HUMAN SKIN AS A ROUTE OF
EXPOSURE TO XENOBIOTICS
• Human skin comes into contact with many toxic agents
such as pesticides and other environmental and
occupational chemicals.

• A multilayered barrier not very permeable.

• The rate -determining barrier is the stratum corneum, the

upper layer of epidermis.
 BIOTRANSFORMATION

• Biotransformation is a metabolic process that takes place mainly in the

liver that facilitates the excretion of both exogenous and endogenous
substances.
• It could also be seen as a process by which substances that enter the
body are changed from hydrophobic to hydrophilic molecules to ensure
elimination from the body.
• However, biotransformations sometimes give rise to toxic metabolites,
via a process called bioactivation. Eg. Biotransformation of
acetaminophen could lead to the formation of a toxic metabolite N-
acetyl-p-benzoquinone imine (NAPQI) which can bind to cellular
macromolecules.
 Classification of
Biotransformation
• Biotransformation process is divided into the following phases:
• Phase I: This involves some reactions such as oxidation, reduction and
hydrolysis to yield a polar, water-soluble, metabolite that could still be
active. The metabolites synthesized in phase I becomes the substrate for
the phase II reactions.
• Phase I reactions are mainly catalyzed by the enzyme, cytochrome P450
system, which is a family of membrane-bound enzymes found within the
endoplasmic reticulum of hepatocytes
• Note: The important biochemical reactions here are: Oxidation, Reduction,
Hydrolysis
 Classification of Biotransformation
Process Continues
• Phase II: Phase II biotransformation process involves the formation a large polar
metabolite by the attachment of endogenous hydrophilic groups to yield water-
soluble inactive compounds that can be excreted by the body.
• The reactions implicated in this phase include
• Methylation:This is carried out by methyltransferases, using S-
adenosylmethionine is methyl group donor
• Glucuronidation (most common): UDP-glucuronic acid is the glucuronyl donor,
and a variety of glucuronosyltransferases, present in both the endoplasmic
reticulum and cytosol, are the catalysts. Molecules such as 2-
acetylaminofluorene, aniline, benzoic acid, meprobamate, phenol, and many
steroids are excreted as glucuronides. The glucuronide may be attached to
oxygen, nitrogen, or sulfur groups of the substrates
• Acetylation: The drug isoniazid, used in the treatment of tuberculosis, is subject to
acetylation
• Sulfation: The sulfate donor in these and other biologic sulfation reactions is
adenosine 3-phosphate-5-phosphosulfate (PAPS)
 Conjugation with glutathione: The enzymes catalyzing these reactions are called
glutathione S-transferases and are present in high amounts in liver cytosol and in
lower amounts in other tissues.
• A variety of glutathione S-transferases are present in human tissue.
• If the potentially toxic xenobiotics were not conjugated to GSH, they would be free to
combine covalently with DNA, RNA, or cell protein and could thus lead to serious cell
damage. GSH is therefore an important defense mechanism against certain toxic
compounds, such as some drugs and carcinogens
 Conjugation with amino acids (glycine and glutamic acid)
REPRESENTATION OF
BIOTRANSFORMATION
 DETOXIFICATION
• The term “detoxification” is sometimes used for many of the
reactions involved in the metabolism of xenobiotics.

• However, it is not appropriate to interchange biotranformation

and detoxification because sometimes biotransformation leads
to the generation of more toxic metabolite in a process known
as BIOACTIVATION. Thus, detoxification should encompass the
process of reducing the toxic load of a particular toxicant
 ROLE OF CYTOCHROME P450 IN
DETOXIFICATION
• The term cytochrome P450 refers to a family of heme proteins present in all
mammalian cell types, except mature red blood cells and skeletal muscle cells, which
catalyze oxidation of a wide variety of structurally diverse compounds.
• Substrates for this enzyme system include endogenously synthesized compounds,
such as steroids and fatty acids (including prostaglandins and leukotrienes), and
exogenous compounds, such as drugs, food additives, or industrial byproducts that
enter the body through food sources, injection, inhalation from the air, or absorption
through the skin.
• Some effects of cytochrome P450 system in medicine include: (i) inactivation or
activation of therapeutic agents; (ii) conversion of chemicals to highly reactive
molecules, which may produce unwanted cellular damage, cell death, or mutations;
(iii) production of steroid hormones; and (iv) metabolism of fatty acids and their
derivatives.
 REACTIONS OF CYTOCHROME
P450
• RH + O2 + NADPH + H+ R OH + H2O + NADP
• RH represents various xenobiotics such as drugs, carcinogens,
pesticides, petroleum products, and pollutants.
• One atom of oxygen enters ROH while the other forma water.
This dual fate of the oxygen accounts for the former naming of
monooxygenases as “mixed function oxidases.
• In the above reaction, the reduced form of cytochrome P450 is
converted to oxidized cytochrome p450.
• Electrons are transferred from NADPH to NADPH-
cytochrome P450 reductase and then to cytochrome P450.
This leads to the reductive activation of molecular oxygen,
and one atom of oxygen is subsequently inserted into the
substrate.
DIAGRAM SHOWING THE MECHANISM OF
CYTOCHROME P450 CATALYZED REACTION
 LOCATION OF CYTOCHROME
P450
 CYP 450 are majorly found the in liver and small intestine but could be found
in all tissues. In liver and most other tissues, they are present mainly in the
membranes of the smooth endoplasmic reticulum, which constitute part of
the microsomal fraction when tissue is subjected to subcellular fractionation.
 In hepatic microsomes, cytochrome P450s can comprise as much as 20% of
the total protein.
 In the adrenal, they are found in mitochondria as well as in the endoplasmic
reticulum.
 The mitochondrial cytochrome P450 system differs from the microsomal
system in that it uses an NADPH linked flavoprotein, adrenodoxin reductase,
and a nonheme iron-sulfur protein, adrenodoxin
INDUCTION OF CYTOCHROME
P450
• The mechanisms of induction of cytochromes P450 could occur
at either the transcriptional or posttranscriptional level,
however, it is not always easy to decipher the mode of a
particular induction based on the inducing compound.
• Induction at transcriptional level has to do with the conversion
of the sequences of genes encoding cytochrome P450 into
mRNA while induction at posttranscriptional level involve ant
process that enhances the level of mRNA transcribed, and this
could involve stabilization of the mRNA or nuclear RNA
processing.
 CLINICAL IMPLICATION OF
CYTOCHROME P450 INDUCTION
• Induction of the cytochrome P450 system may result in altered efficacy of therapeutic
drugs, as the accelerated rate of hydroxylation will increase the inactivation and/or
enhance the excretion rate of drugs. Induction may also cause unexpected and
unwanted side effects of therapeutic agents due to increased formation of toxic
metabolites that may cause cell injury if produced in large enough concentrations.
• For example, consider the circumstance where a patient is taking the anticoagulant
warfarin to prevent blood clotting. This drug is metabolized by CYP2C9.
Concomitantly, the patient is started on phenobarbital (an inducer of this P450) to
treat a certain type of epilepsy, but the dose of warfarin is not changed. After some
days, the level of CYP2C9 in the patient’s liver will be elevated three- to fourfold.
• This indicates that warfarin will be metabolized much more quickly than before, and
its dosage will have become inadequate. Therefore, the dose must be increased if
warfarin is to be therapeutically effective. In the same vein, a problem could arise
later on if the phenobarbital is discontinued but the increased dosage of warfarin
stays the same.
 CLINICAL IMPLICATION CONTINUES

• The patient will be at risk of bleeding, since the high dose of warfarin
will be even more active than before, because the level of CYP2C9 will
decline once phenobarbital has been stopped.
• Another example of enzyme induction involves CYP2E1, which is
induced by consumption of ethanol. It is important to note that
CYPP450 metabolizes various solvents and components found in
tobacco smoke, of which some are carcinogens.
• Hence, if the activity of CYP2E1 is elevated by induction, this may
increase the risk of carcinogenicity developing from exposure to such
compounds.
 EXCRETION OF XENOBIOTICS
• The primary excretion/elimination routes
• Urinary-Renal
• Fecal
• Respiratory- Exhalation
• Others
 URINARY-RENAL EXCRETION
• Renal excretion is made up of 3 distinct processes namely:
Glomerular filtration
o Molecular weights larger than 60 kDa and plasma protein
bound xenobiotics can not be filtered at the glomeruli.
o Presence of albumin or blood cells in urine is the
indication of glomerular damage.
 Active Tubular Secretion

Xenobiotics could be excreted into the urine by active

secretion at proximal tubule of the nephron.

Protein bound xenobiotics, polar metabolites and

conjugates are readily secreted by this mechanism.

 Secretion occurs by active transport mechanisms

 TUBULAR EXCRETION-
REABSORPTION
• Xenobiotics can be reabsorbed from urine into blood capillaries via
passive diffusion at distal tubule of the nephron.
• Urine pH greatly affects reabsorption or urinary excretion. Hence, by
altering the pH of the urine xenobiotic reabsorption back to
bloodstream or remain in the tubular lumen and excreted with urine
could be achieved.
• If the urine is alkaline, acidic drugs (xenobiotic) are more ionized and
less lipophilic, and hence excreted more readily. In acidic urine,
alkaline drugs (xenobiotic) are readily excreted. Thus, acidic urine
causes reabsorption of acidic drugs which reduces renal excretion.
 FECAL EXCRETION
• Biliary Excretion
• In liver xenobiotics as metabolites or conjugates are transferred
into the bile and thereafter the intestine.
• Xenobiotics can either be excreted with feces or undergo an
enterohepatic circulation.
• This method is regulated by active transport. e.g. Organic bases
and acids, neutral substances, and metals (As, Pb, Hg) are
excreted by biliary route
 Entero-Hepatic Circulation
• Hydrolytic enzymes in the intestinal flora can hydrolize
polar glucuronide and sulfate conjugates making them
lipophilic for reabsorption.
• Hence, reabsorption takes place and xenobiotic returns to
the liver again by the portal vein.
• This repeated process is called enterohepatic circulation
• Half-life of a chemical is prolonged and toxicity can be
enhanced.
 RESPIRATORY EXHALATION
• The lungs are also an important excretion route for
xenobiotics and their metabolites in the gaseous phase of
blood.
• Elimination of gases is normally inversely proportional to
the rate of their absorption.
• Xenobiotics are eliminated by passive diffusion. e.g.
Ethanol, anesthetic gases
 FECAL EXCRETION
• In some cases, some xenobiotics can be eliminated via the
feces by direct intestinal excretion.

• This is a relatively slow process.

• It is the major elimination route for compounds that have

low rates of biotransformation and/or low renal or biliary
clearance.
THANKS