Toxicokinetics & Toxicodynamics

 Toxicokinetics (Determines the no. molecules that

can reach the receptors)
• Uptake
• Transport
• Metabolism & transformation
• Sequestration
• Excretion
 Toxicodynamics (Determines the no. of receptors
that can interact with toxicants)
• Binding
• Interaction
• Induction of toxic effects
 Uptake and Elimination

K1 K2
Biological Elimination
Uptake
System

K1 > K2 : Accumulation & Toxic effect

 Toxicokinetics
1. Uptake
2. Transport
3. Metabolism & Transformation
4. Sequestration
5. Excretion
 Uptake routes
1. Ingestion (toxicity may be modified by
enzymes, pH and microbes)
2. Respiration (Air borne toxicants)
3. Body surface (Lipid soluble toxicants
such as carbon terta chloride and
organophosphate)
 Uptake Barriers

1. Cell membrane
2. Cell wall/cuticles/stomata
3. Epithelial cells of GI tract
4. Respiratory surface (lung, gill
tracheae)
5. Body surface
 Uptake of Toxicants

1. Passive diffusion
2. Facilitated transport
3. Active transport
4. Pinocytosis
 Uptake by Passive diffusion
 Uncharged molecules may diffuse along
conc. gradient until equilibrium is reached
 Not substrate specific
 Small molecules of < 0.4 nm (e.g. CO, N20,
HCN) can move through cell pores
 Lipophilic chemicals may diffuse through
the lipid bilayer
 Uptake by Passive diffusion
 First order rate process, depends on:
– Concentration gradient
– Surface area (aveoli = 25 x body surface)
– Thickness (fluid mosaic phospholipid bi-layer
ca. 7 nm)
– Lipid solubility & ionization(dissolved before
transport, polar chemicals have limited
diffusion rate)
– Molecular size (membrane pore size = 4-40 A,
allowing MW of 100-70,000 to pass through)
 Diffusion governed by Flicks law
D/dt = KA (Co - Ci) / X
 Where:
– dD/dt = rate of transport accross the membrane
– K= constant
– A= Cross sectional area of membrane exposed to the
compound
– Co = Concentration of the toxicant outside the
membrane
– Ci = Concentration of the toxicant inside the
membrane
– X= Thickness of the membrane
Uptake by Facilitated Transport
 Carried by trans-membrane carrier along
concentration gradient
 Energy independent
 May enhance transport up to 50,000 folds
 Example: Calmodulin for facilitated
transport of Ca
 Uptake by Active Transport
 Independent of or against conc. gradient
 Require energy
 Substrate –specific
 Rate limited by no. of carriers
 Example:
– P-glycoprotein pump for xenobiotics (e.g. OC)
– Ca-pump (Ca2+ -ATPase)
 Uptake by Pinocytosis
 For large molecules ( ca 1 um)
 Outside: Infolding of cell membrane
 Inside: release of molecules
 Example:
– Airborne toxicants across alveoli cells
– Carrageenan accross intestine
 Transport & Deposition
 Transport
• Blood
• Lymph, haemolymph
• Water stream in xylem
• Cytoplamic strands in phloem
 Deposition
Toxicant Target organs
Pb Bone, teeth, brain
Cd Kidney, bone, gonad
OC, PCB Adipose tissue,milk
OP Nervous tissue
Aflatoxin Liver
 Metabolism & Transformation
 Evolved to deal with metabolites and
naturally occurring toxicants
 Principle of detoxification:
1. Convert toxicants into more water
soluble form (more polar & hydrophilic)
2. Dissolve in aqueous/gas phases and
eliminate by excretion (urine/sweat) of
exhalation
3. Sequestrate in inactive tissues (e.g bone,
fat)
 P450 system
 A heme-containing cytochrome protein
located in ER, and is involved in electron
transport.
 Highly conservative, occur in most plants
& animals
 Two phases of transformation
 May increase or decrease toxicity of
toxicants after transformation (e.g turn
Benzo[a]pyrene into benzo[a]pyrene diol
epoxide, and nitroamines into methyl
radicals)
 Inducible by toxicants
 Induction of P450
Toxicant

Aryl Hydrocarbon
Receptor

Toxicant-Receptor
Complex Bind at
Specific site hours

Translocating
protein
m-RNA for CYP1A
 Phase I Transformation
 Mixed Function Oxidase (MFO) System in
smooth ER is responsible (Microsomes)
 In vertebrates, primarily found in liver
parenchyma cells, but also other tissues
(e.g intestine, gill)
 In invertebrates, found in hepatopancrease
& digestive glands
 Lower MFO activities in molluscs
 Add polar group(s) to increase
hydrophilicity for Phase II transformation
 Examples of Phase I Transformation
 Hydrolysis

RCOO-R’ + H2O ---------> RCOO-H + R’-OH

 Hydroxylation

NADP NADP+
R-H --------------------------> R-OH + H2O
 Examples of Phase I Transformation

 Epoxidation
O
R-CH==CH-R’ -----------> R---CH ----CH-R’
Phase II transformation
• Cytochrome P450 II enzyme systems
in cytosol is responsible
• Covalent conjugation to water soluble
endogenous metabloites (e.g. sugars,
peptides, glucuronic acid, glutathione,
phosphates & sulphate)
• May involve deamination, acyclic
hydroxylation, aromatic hydroxylation,
and dealkylation
• Further increase hydrophilicity for
excretion in bile, urine and sweat
 Important Phase II enzymes

 Glutathion S-transferases (GST)

 Epoxide Hydrolase (EH)
 UDP-glucuronosyltransferase (UDP-
GTS)
 Sulfotransferase (ST).
 Examples of Phase II Transformation

 Deamination

R-NH2 ---------------------------> R=O + NH3

Examples of Phase II Transformation
 Dealkylation

R-CH2-CH3 ----------------------> R + CH3-CH2O

 Dehalogenation:

R-Cl ---------------------------------> R-H + Cl+

Glutathione-S-transferase (GST)

O GST
R------R’ ----------------------> HO-R-SG

GST
R-Cl ------------------------------> R-SG + Cl
 Sequestration
 Animals may store toxicants in inert tissues
(e.g. bone, fat, hair, nail) to reduce toxicity
 Plants may store toxicants in bark, leaves,
vacuoles for shedding later on
 Lipophilic toxicants (e.g. DDT, PCBs) may
be stored in milk at high conc and pass to
the young
 Metallothionein (MT) or phytochelatin
may be used to bind metals
 Excretion
 Gas (e.g. ammonia) and volatile (e.g. alcohol)
toxicants may be excreted from the gill or lung
by simple diffusion
 Water soluble toxicants (molecular wt. < 70,000)
may be excreted through the kidney by active or
passive transport
 Conjugates with high molecular wt. (>300) may
be excreted into bile through active transport
 Lipid soluble and non-ionised toxicants may be
reabsorbed (systematic toxicity)
 Tutorial Questions
1. Find TWO enzymes/proteins which are
inducible by xenobiotics or metals
2. Molluscs have low P450 activities. They are
often used as pollution indicators for metals
and xenobiotics. Explain why.
3. Lipophilic compounds may normally have a
longer biological half life. Explain why.
4. Why exposure of animals to sub-lethal level of
toxicants may increase tolerance of the
organisms to the chemical.