Biopharmaceutical Technology

Faculty Name : Mr.Sagar Nagare

Lecture 11

Bioactivation and tissue

toxicity
Bioactivation

Definition:-Bioactivation is defined as: Enzymatically formed metabolites,

which are more reactive than the mother substance and excreted metabolites.
OR
Formation of highly reactive metabolites (from relatively inert chemical
compounds) which interact with the tissues to precipitate one or more of the
several forms of toxicities such as carcinogenesis is called as bioactivation or
toxicological activation.
Tissue toxicity:
The most significant toxicological effects of xenobiotics are reactive metabolites are:-
 can react with nucleophilic sites
 SH groups (glutathione, cysteine)
 NH2 and – COOH groups (DNA, RNA, proteins)

Lecture 11 bioactivation and tissue

toxicity
Bioactivation

Imbalance between formation and detoxification of reactive metabolites

can arise from:
 enzyme induction (increased biotransformation and formation of reactive
metabolites)
 high dose of xenobiotic
 depletion of cellular defence mechanisms.
 saturation of non-toxic pathways

Lecture 11 bioactivation and tissue

toxicity
Imbalance between formation and detoxification of reactive metabolites
can arise from:
 enzyme induction (increased biotransformation and formation of reactive
metabolites)
 high dose of xenobiotic
 depletion of cellular defence mechanisms.
 saturation of non-toxic pathways

Lecture 11 bioactivation and tissue

toxicity
MECHANISM OF TISSUE TOXICITY

Lecture 11 bioactivation and tissue

toxicity
 Examples of bioactivating compounds

Compounds Reactive pathway or Factors increasing

intermediate product toxicity
Acetaminophen N-hydroxylation Sulphate and GSH
depletion
Acetyl hydrazine N-hydroxylation
Aflatoxin B Epoxidation Further metabolism
Benzene Epoxidation
Benzo[a]pyrene Epoxidation
PCB Epoxidation GSH depletion
Tetrachlorcarbon Free radicals Reductive metabolism
Halothane Free radicals Reductive metabolism
Parathion Oxidation with
sulphur formation

Lecture 11 bioactivation and tissue

toxicity
The reactive, chemically unstable species, capable of toxification,
are broadly divided into two categories:
 Electrophiles
 Free radicals

Electrophiles: - are species deficient in electron pair. The enzyme system

through which they generated is cytochrome P-450. Carbon, nitrogen or
sulphur containing compounds can be metabolically activated to yield
electrophiles.

 Important electrophiles are:

 Epoxides;-e.g., epoxide of benzo(a)pyrene present in cigarette smoke
which causes cancer.
 Hydroxylamines, nitroso and azoxy derivatives, nitrenium ions and
elemental sulphur.

Lecture 11 bioactivation and tissue

toxicity
Mechanism:-

 The mechanism by which electrophiles precipitate toxicity is through

covalent binding to nucleophilic tissue components such as
macromolecules(proteins, nucleic acids, and lipids) or low molecular weight
cellular constituents.
 Covalent binding to DNA is responsible for carcinogenicity and tumour
formation.
The body’s defence against electrophiles is their inactivation by conjugation
with glutathione, the most abundant cellular nucleophile with –SH group.
An e.g. of tissue toxicity due to electrophiles is hepatotoxicity of paracetamol
metabolites

Lecture 11 bioactivation and tissue

toxicity
HEPATOTOXICITY OF PARACETAMOL METABOLITES

Lecture 11 bioactivation and tissue

toxicity
Free radicals

 Free radicals are species containing an odd number of electron. They may
be positively charged (cation radical), negatively charged (anion radical) or
neutral radical.
R+ R- R
Cation radical Anion radical neutral radical

 Free radicals are generally formed via NADPH cytochrome P-450

reductase or other flavin containing reductases.
 Xenobiotics that on metabolic activation yield free radicals are Quinone’s,
aryl amines, nitro aryls and carbon tetrachloride.
 Endogenous compounds such as epinephrine and DOPA can also generate
free radicals.
 Most free radicals are organic.
 They provide toxicity by peroxidation of cellular components.

Lecture 11 bioactivation and tissue

toxicity
Free radicals

•An important class of free radicals is inorganic free radicals such as hydrogen
peroxide and superoxide anion.
•These oxidative moieties can cause tremendous tissue damage leading to
mutations or cancer.
•The potential toxicity of free radicals is far greater than that of the
electrophiles.
•Cellular defence mechanisms against free radicals include control imposed by
membrane structure, neutralization by glutathione, control exerted by non-
enzymatic antioxidant scavengers such as vitamin A, E and C and enzymatic
inactivation of oxygen derived free radicals.
• Generation of reactive metabolites is indicated by modification in enzyme activities,
formation of glutathione conjugates and depletion in tissue levels of glutathione.
Since the availability of glutathione in the body determines the threshold for toxic
response, thiols(e.g., N-acetyl cysteine) can be used to treat poisoning by drugs
such as paracetamol that yield reactive metabolites.

Lecture 11 bioactivation and tissue

toxicity
Compounds and their metabolic reactions that generate toxic
intermediates

Benzo(a)pyrene Aromatic epoxidation Lung cancer

Aflatoxin B1 Olefin epoxidation Hepatic cancer
Thalidomide Hydrolytic cleavage of Teratogenesis
lactam
Chlorinated Oxidative Nephrotoxicity
hydrocarbons dehalogenation

Lecture 11 bioactivation and tissue

toxicity
Reference

BIOPHARMACEUTICS AND PHARMACOKINETICS BY

D.M.BRAHMANKAR,SUNIL B.JAISWAL.P.NO-189-191
www.sciencedirect.com
Biopharmaceutics & Pharmacokinetics by Milo Gibaldi, 4th
edition

Lecture 11 bioactivation and tissue

toxicity
Thank You