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Xenobiotic Biotransformation
Michael A. Trush, PhD
Johns Hopkins University
Section A
Biotransformation: Basic Concepts
Glomerulus
Eerent arteriole
Bowmans capsule
Unfiltered drug
Passive
reabsorption
Filtered drug
Proximal tubule
Active
secretion
Water soluble
Lipid soluble
Biotransformation of Xenobiotics
Biological basis for xenobiotic metabolism:
w To convert lipid-soluble, non-polar, nonexcretable forms of chemicals to watersoluble, polar forms that are excretable in
bile and urine
Continued
Biotransformation of Xenobiotics
Biotransformation Reactions
w Phase I Reactions
Enzymatic reactions that add or
expose functional groups to xenobiotics
such as -OH, -SH, -NH2
or COOH
Functional groups are analogous to
having a trailer hitch on a vehicle
Continued
Biotransformation Reactions
w Phase II Reactions
Enzymatic reactions that result in the
conjugation of large water-soluble,
charged (polar)biomolecules to
xenobiotics
For these reactions to occur, a
functional group must be present on
either the parent compound or its
Phase I product
8
TRUCK
lipophilic
not charged
not water soluble
poorly excretable
Photo by John Pittman. Creative Commons BY-NC-SA.
Phase 1 enzymes
add or expose a
functional group
HITCH
TRUCK
lipophilic
not charged
not water soluble
poorly excretable
still lipophilic
possibly reactive
poorly water soluble
poorly excretable
catalyzed by P450s
10
Phase 1 enzymes
add or expose a
functional group
HITCH
TRUCK
lipophilic
not charged
not water soluble
poorly excretable
still lipophilic
possibly reactive
poorly water soluble
poorly excretable
catalyzed by P450s
Phase 2 enzymes
conjugate (transfer)
endogenous molecules*
to the functional group
TRAILER
not lipophilic
usually not reactive
water soluble products
excretable
catalyzed by transferases
Section B
Biotransformation: Enzymes
Cell(s)
Parenchymal cells (hepatocytes)
Proximal tubular cells (S3 segment)
Clara cells, Type II alveolar cells
Mucosa lining cells
Epithelial cells
Seminiferous tubules, Sertolis cells
14
15
Cytochrome P450
Characteristics
w Can metabolize many xenobiotics (broad
substrate specificity)
w Can catalyze many types of reactions
w Is widely distributed among tissues, and
tissue distribution can be quite varied
Continued 16
Cytochrome P450
Characteristics
w Exists in multiple forms (determined by
different genes)
w Levels can be increased by exposure to
chemicals in the food, water, or air
(induction)
17
Continued 18
19
20
21
Phase II Reactions
w Many of the characteristics described
earlier for cytochrome P450 also apply to
these Phase II enzymes
w However, cytochrome P450 is localized in
cellular membraness, whereas Phase II
enzymes are, for the most part, in the
cytoplasm (water portion) of cells
22
23
Glutathione-S-Transferase
Structure of Reduced Glutathione (MW 307)
24
Glutathione-S-Transferase
w Glutathione adducts are further metabolized in
the kidney to derivatives referred to as
mercapturic acids of the associated xenobiotic.
This occurs in the kidney.Mercapuric derivatives
are then found in the urine.
w Glutathione adducts are excreted in the bile and
feces un-changed.
w Some chemicals are reactive enough to form
glutathione adduct without the assistance of GSH
transferase.
25
Section C
Factors Affecting Biotransformation
Continued 27
28
Mouse
Rabbit
Rat
Dog
Duration of
Action
Relative Enzyme
Activity
(g hexobarbitol
metabolized/gm/liver/hr)
12 8
49 12
90 15
315 105
598 184
196 28
134 51
36 30
Source: G.P. Quinn, et.al. Species, Strain and Sex Differences in the Metabolism of
Hexobarbital, Aminopyrine and Aniline. Biochem. Pharmacol. 1:152, 1958
30
Species, Strain,
and Genetic Variation
Continued 31
Species, Strain,
and Genetic Variation
32
His+ revertants/mg
Metabolism of Aflatoxin B1
and Benzo[a]pyrene
4000
3000
2000
1000
A B CDE FG H I J
A B CDE FG H I J
Biopsy samples
50
40
30
20
10
0
9 10 11 12
34
Birth
Puberty
Adult
Age
Schematic representation of the ontogeny of
hepatic drug metabolic activity
35
36
37
Section D
Induction of
Biotransformation Enzymes
Induction of Xenobiotic
Metabolizing Systems
1. Many chemicals can induce the synthesis
of the enzymes involved in Phase I and II
xenobiotic metabolism and include
chemicals found in the environment, the
diet, and cigarette smoke
2. Inducers often exhibit specificity for the
enzymes which they induce
Continued 39
Induction of Xenobiotic
Metabolizing Systems
3. Depending on the inducer, fairly high
dose levels or repeated dosing may be
required; on the other hand, TCDD
(dioxin) is effective as an inducer at
1 microgram/kg in some species
Continued 40
Induction of Xenobiotic
Metabolizing Systems
4. Studies have demonstrated that a
cluster of genes referred to as the
Ah locus controls the induction of
xenobiotic enzyme activities by polycyclic
aromatic compounds and TCDD
Continued 41
Induction of Xenobiotic
Metabolizing Systems
5. Such toxic responses as cancer, chemicalinduced cataracts, aplastic anemia, and
fetal toxicity have been demonstrated to
be affected by this cluster of genes
6. Evidence exists for the Ah locus in man
42
Phenobarbital
Polycyclic
Hydrocarbons
Increase
No effect
Increase
No effect
Increase
No effect
Increase
Increase
Small increase
No effect
No effect
Increase
Increase
No effect
43
Polycyclic
Hydrocarbons
812 hours
36 hours
35 days
2448 hours
57 days
512 days
Marked
Slight
Large increase Small increase
Marked increase No effect
Increase
No effect
Increase
No effect
Increase
Small increase
Small increase Small increase
Increase
Small increase
None identified Identified
Phenobarbital
Continued 44
The Ah Receptor
w Ah receptor = Arylhydrocarbon receptor
w Examples = 3-methylcholanthrene
benzo[a]pyrene
w Also called TCDD receptor or dioxin
receptor
Continued 45
nucleus
cytoplasm
46
Section E
Bioactivation and Toxicity
48
49
50
51
Chemical Nature of
Reactive Intermediates
w ElectrophilesForm covalent (irreversible)
bonds with cellular nucleophiles such as
GSH, proteins and DNA
w Free RadicalsOdd or unpaired electron
Can act as electrophiles
Can abstract hydrogen from target
molecules, such as lipids or nucleic acids
Can activate molecular oxygen
52
53
54
Bioactivation of Acetaminophen
100
Glutathione
80
60
1
40
20
Covalent binding
20
40
60
100
200
400 600
1000
55
56
57
58
59
61