Xenobiotic metabolism

INTRODUCTION TO TOXICOLOGY AND TOXICOLOGICAL

CHEMISTRY
Most pollutants and hazardous substances are of concern for their
potential to harm living organisms
Toxicology
Toxicology is the science of poisons
A poison or toxicant is a substance that is harmful to living organisms
because of its detrimental effects on
Tissues Organs Biological Processes
 MajorSitesofExposure,Metabolism,andStorage,Routesof
DistributionandEliminationofToxicSubstancesfromtheBody

Ingestion(entrysite) Gastrointestinaltract

Inhaledair Feces
(excretion)
(entrysite) Bile
Dermalexposure
Portal (entrysite)
Pulmonarysystem vein Liver
(lungandalveoli)

Skin

Exhaledair
(excretion)
Bloodandlymphsystem

Protein Metabolism Distributionoffree,

binding bound,ormetabolite
form

Cell membrane Kidney

Receptorcells
Bone Bladder
Fat
Toxicant Urine
storage (excretion)
Exposurefactors
Dose Toxicantconcentration Duration
Frequency Rate Site Route
Acute Chronic
systemic systemic

Acute Chronic
local local
Timeofexposure

Additiveandsynergisticeffectsfortoxicantswiththesamefunction
Potentiationfromaninactivesubstance,antagonismfromanother
activesubstance
 DoseResponseRelationships
Doseistheamount(perunitbodymass)oftoxicanttowhichthe
organismisexposed
Responseistheresultingeffectonanorganism
100

Percent
deaths
50

LD50

0
Logdose
 RELATIVE TOXICITIES
SubstanceApproximateLD50*Toxicityrating
5
105 1.Practicallynontoxic
DEHP 4 >1.510 4mg/kg
Ethanol 10 2.Slightlytoxic,510 3
Sodiumchloride 3
Malathion to1.5104 mg/kg
10 3.Moderatelytoxic,
Chlordane 2 500to5000mg/kg
Heptachlor 10
4.Verytoxic,50to
Parathion 500mg/kg
10
5.Extremelytoxic,
TEPP 1 5to50mg/kg

Tetr odotoxin# 10 1
Inlandtaipanvenom
10 2 6.Supertoxic,
3 <5mg/kg
TCDD5 10

10 4

Botulinustoxin 10 5

*LD50valuesareinunitsofmgoftoxicantperkgofbodymass.
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paralysisandensuingrespiratoryfailure);but,doubtlessly,thebiggestthrillisthestrikinghubrisofthe
event,theostentatiousrisktakingthatinvolvessomuchforsolittle.
 ReversibilityandSensitivity
Exposureto Reversible
toxicant

Irreversible
Effect

Sublethal Lethal
effect Marginofsafety
effect

Hyposensitivity
Normal
Hypersensitivity
XENOBIOTIC AND ENDOGENOUS SUBSTANCES
Xenobiotic substances are foreign to living systems
Endogenous substances occur naturally in biological systems
Figure 21.4. Organisms require acceptable levels of endogenous
substances and may respond adversely to deficiencies or excess
 What is xenobiotic?

A xenobiotic (Gk xenos stranger) is a compound that is

foreign to the body.
The principal classes of xenobiotics of medical relevance are
drugs, chemical carcinogens, and various compounds that
have found their way into our environment
More than 200,000 manufactured environmental
chemicals exist (including heavy metal).
Most of these compounds are subject to metabolism
(chemical alteration) in the human body, with the liver being
the main organ involved;
occasionally, a xenobiotic may be excreted unchanged.
TOXICOLOGICALCHEMISTRY
Toxicologicalchemistryisthesciencethatdealswiththechemical
natureandreactionsoftoxicsubstances,includingtheirorigins,uses,
andchemicalaspectsofexposure,fates,anddisposal.

Toxicant ..... +..... Organism Toxic

effect

Toxicologicalchemistry Toxicology

Organismsmetabolizexenobioticspeciesbyenzymecatalyzed
PhaseIandPhaseIIreactions.
Quantitativestructureactivityrelationships(QSAR)relatethe
chemicalnatureofsubstancestotheirbiochemicalreactions.
 Most Toxic Substances and their Precursors
(Protoxicants) are Metabolized, Commonly by
Phase I and Phase II Reactions
Designed to detoxify
Product is usually more readily eliminated
May produce a toxic or more toxic form
 Biotransformation
Water soluble xenobiotics are easier to eliminate
( t1/2)
Urine, feces but not exhalation
If within barrier, no out
Multiple enzymes (families)
Constitutively expressed
Inducible
Broad specificity
Polymorphic (allelic variants)
Stereo-isomer specificity: 6-OH in hormones:
CYP2A1 6-OH
CYP3A 6-OH
 Biotransformation

Potentially toxic xenobiotic Relatively harmless

Metabolic
Detoxification activation

Inactive metabolite Reactive intermediate

Converting lipophilic to water
soluble compounds
Lipophilic
Xenobiotic (non-polar)
Phase I - Activation

Reactive intermediate

Phase II - Conjugation

Conjugate
Water soluble
Excretion (polar)
 Phase I

introduction of functional group

hydrophilicity increases slightly
may inactivate or activate original compound
major player is CYP or mixed function
oxygenase (MFO) system in conjunction with
NAD(P)H
location of reactions is smooth endoplasmic
reticulum
 Phase II

conjugation with endogenous molecules

(GSH, glycine, cystein, glucuronic acid)

hydrophilicity increases substantially

neutralization of active metabolic intermediates
facilitation of elimination
location of reactions is cytoplasm
 Phase I Reactions, Figure 21.6

Most Phase I reactions are microsomal mixed-function oxidase

Reactions
Catalyzed by the cytochrome P-450 enzyme system
Associated with cell endoplasmic reticulum
Occurring most abundantly in the liver of vertebrates
 Phase I reactions Table 6.1

Oxidation
Hydroxylation (addition of -OH group)
N- and O- Dealkylation (removal of -CH side chains)
Deamination (removal of -NH side chains) O epoxide
Epoxidation (formation of epoxides) C C
Oxygen addition (sulfoxidation, N-oxidation)
Hydrogen removal

Reduction
Hydrogen addition (unsaturated bonds to saturated)
Donor molecules include GSH, FAD, NAD(P)H
Oxygen removal

C O
Hydrolysis
Splitting of C-N-C (amide) and C-O-C (ester) bonds

See also Chapter 6 of Casarett and Doulls Toxicology

 Biotransformation
Activation of xenobiotics is a key element
(e.g. benzene, vinyl chloride)
Reactive intermediates include epoxides and free
radical species (unpaired electrons) that are short-lived
and hence highly reactive
Protection is provided by
endogenous antioxidant substances, e.g. GSH
vitamins C and E
antioxidant enzymes, SOD, GPX, CAT in coupled reactions
Antioxidant molecules are oxidized in the process but
have the capacity to regenerate the reduced form from
the oxidized - NAD(P)H is a key player

See also p. 40-44 of Casarett and Doulls Toxicology

 Cytochrome P450 (CYP)
Mixed Function Oxidases (MFO)
Located in many tissues but highly in liver ER
Human: 16 gene families
CYP 1,2,3 perform drug metabolism
>48 genes sequenced
Key forms: CYP1A2, CYP2C9, CYP2C19, CYP2D6,
CYP2E1, and CYP3A4
Highly inducible
Alcohol CYP2E1
Dioxin/PCBs CYP1A
Barbiturates CYP2B
CYP genes have multiple alleles (2D6 has 53, and 2E1 has
13)
The CYP-450 reaction cycle
A

G
(B)

D
Oxidation of vinyl chloride to an
epoxide
 Metabolic enzymes
1. Microsomal:
1. CYP450 monooxygenases
2. Flavin monooxygenase
2. Non-microsomal
1. Alcohol dehydrogenase
2. Aldehyde dehydrogenase
3. Monoamine and diamine oxidases
3. Both
1. Esterases and Amidases
2. Prostaglandin synthase
3. Peroxidases
At least30 different enzymes catalyze reactions involved
inxenobiotic metabolism
Cooxidation of
acetaminophen
by prostaglandin
endoperoxide
synthetase

Compare to fig. 6-2

Hydrolysis of esters and amides
Hydrolysis of organophosphates
Hydrolysis of epoxides
Stereoselective
hydroxylation
Metabolism of
benzo(a)pyrene to
9,10 epoxide:
Potent mutagen
that binds DNA
Azo- and nitro- reduction
 Intestinal flora as part of biotransformation

Ready for elimination

n Flora action
tio
r p
o
a bs
re
Oxidation reactions
 Benzene trasformation to
leukemia-causing metabolite
Flavin mono-oxygenases
(FMO) catalyzed reactions

Nitrogen compounds
 The term detoxification is sometimes used
formany of the reactions involved in the
metabolism of xenobiotics.
However, the term is not always appropriate
because, as mentioned above, in some cases the
reactionsto which xenobiotics are subject
actually increase their biologic activity and
toxicity.
 Phase II Reactions, Figure 21.7
O
Carboxyl: C O Endogenous
+ conjugating
Xenobiotic Hydroxyl: OH agent
compound,
Halogen: F,Cl,Br,I
oftenPhase
1reaction C
Epox ide : O
product Conjugation
C
H product
Amino: N
Higherpolarity
H
Functionalgroupsthatr eact Greaterwater
withaconjugatingagent
O solubility
Moreeasily
Glucuronideconjugate C OH eliminated
O
OH X R
HO
OH
 Phase II reactions
Glycoside conjugation - glucuronidation
Sulfate - sulfation
Glutathione (GSH)
Methylation
Acylation
Acetylation
Amino acid conjugation
Deacetylation
Phosphate conjugation
Glucuronidation of phenol
Sulfation of phenol and toluene
GSH conjugation of
acetaminophen
 Glutathione

-glutamyl-cysteinyl-glycine

Active site of a GST:

 Mention some toxic metal and its effect on
health?
Toxic heavy metals such as lead, cadmium and mercury can
disrupt brain chemistry and human functioning.
 How Lead affects Health:
Bones - Instead of calcium, lead is incorporated into bone.
Brain - Lead can inhibit copper-dependent enzymes needed for
neurotransmitters, causing hyperactivity.
Energy - Fatigue is triggered by increasing rate of destruction of
red blood cells. Also, lead inhibits copper and iron-dependent
enzymes in the Krebs cycle.
Kidneys - Gout can occur from lead toxicity raising uric acid
levels and impairing kidney functions.
Minerals - Lead inhibits calcium, zinc, manganese, copper, and
iron causing deficiencies.
Thyroid gland - Lead can inactivate the thyroid hormone thyroxin
because it interferes with the iodine uptake to the thyroid gland.
 How Mercury affects health:
Kidneys - mercury accumulates in the kidneys causing kidney
damage
Energy - mercury compounds inhibit the enzyme ATPase,
which impairs energy production in all body cells.
Nervous system - degeneration of nerve fibers and reduced
motor conduction speed.
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