HLTH 340

Lecture A2
Toxicokinetic processes:
absorption (part-1)

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HLTH 340 Lecture A2

Basic Steps in Toxicological Analysis

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HLTH 340 Lecture A2

Toxicokinetic processes- also termed

pharmacokinetics, ADME, disposition

toxicokinetics describes the movement of xenobiotic substances into and within the
organism subsequent to an environmental exposure

descriptive (semi-quantitative) analysis

quantitative analysis (mathematical formulas and graphs)
computer-based simulations (PB-PK models = physiologically-based pharmacokinetic models)

Absorption controls entry of xenobiotics through the external membrane barriers into the
blood (or lymphatic) circulation

local effect (tissues near site of absorption)

regional effect (tissues downstream from site of absorption -- first-pass effects
systemic effect (throughout the body)

Distribution determines the movement of xenobiotic molecules with the circuatory fluids
and specific organs and tissues

Metabolism (biotransformation) describes the biochemical processes that convert the

original (parent) xenobiotic to various metabolic products (metabolites)

Excretion controls the removal of the xenobiotic or its metabolites from the body
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Toxicokinetic (ADME) processes

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Toxicokinetic and toxicodynamic

pathways jointly affect toxicity

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Route of exposure
Route of exposure

The ROUTE (site) of exposure is an important

determinant of the ultimate DOSE different routes
may result in different rates of absorption.

Dermal (skin)

Inhalation (lung)

Oral (GI)

Injection

The ROUTE of exposure may be important if there

are tissue-specific toxic responses.

Toxic effects may be local (in a specific tissue) or

systemic (throughout the organism)

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Routes of Absorption, Distribution and Excretion

Ingestion

Inhalation

Intravenous

Intraperitoneal
Subcutaneous

Gastrointestinal
tract

Lung

Intramuscular

first-pass
effect

Dermal

Liver

distribution

Blood and lymph

Bile

extracellular
fluid

Kidney

Bladder

feces

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absorption

Urine

Lung

Secretory
Structures

Expired Air

HLTH 340 Lecture A2

body
organs

soft
tissue

Alveoli

Secretions

fat

bone

excretion
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First-pass extraction: the hepatic portal vein carries

absorbed nutrients and xenobiotics to the liver

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HLTH 340 Lecture A2

Absorption of molecules across

external and internal membrane barriers
passive diffusion

receptor-mediated transport
(selective)

(non-selective)

transcellular
paracellular

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Types of membrane transport mechanisms:

active transport and passive transport

external dose
(site of
absorption)

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internal
dose
(blood)

HLTH 340 Lecture A2

external dose
(site of
absorption)

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Intestinal absorption via passive diffusion using

paracellular and transcellular permeation pathways

intercellular tight junction

(can be open, closed, or leaky

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Paracellular permeation through a membrane barrier

occurs between adjacent cell membranes
The characteristics of the paracellular pathway are
defined by specific junctional complexes that span
the intercellular space. There are four types of
complexes:
(1)
(2)
(3)
(4)

apical
(outside)

zona occludens, or tight junction;

zona adherens, or intermediate junction;
desmosomes; and
gap junctions.
Specific proteins localized to each complex link
adjacent cells and the cytoskeleton. Original models
of the paracellular pathway as a static barrier are
being replaced by a more dynamic model in which
the junctional complexes are involved in signaling
and regulation, most likely through protein
phosphorylation or dephosphorylation.
The tight junction is the most apical complex and is
believed to control permeability across the
paracellular pathway through a series of strands and
grooves. Molecular definition of the specific
components of the tight junction ( eg , Z0-1, Z0-2,
occludin, cingulin) may permit a clearer
understanding of how the tight junction functions as
a barrier
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baso-lateral
(inside)

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The tight junction (TJ) barrier structure forms pore

structures between adjacent cell membranes
claudins

The TJ barrier consists of two components

physiological pores and pathological
breaks.
All epithelial TJs have a system of small
approximately 8-angstrom pores that varies
among cell types in ionic charge selectivity and in
porosity, i.e. the apparent number of pores.
The mechanism controlling overall porosity
is unclear, but it is known that preferences for ionic
charges is controlled by claudins.
The claudins form the pore structure
or influence their size and shape.
Each claudin has a characteristic
influence on the permeability for small
cations and anions.
The passage of material larger than approximately
8-angstroms shows no charge selectivity. This
small pathway may represent a pathological break
between cells. Such disruptions can arise in
response to proinflammatory factors like
interferon-gamma and tumor necrosis factoralpha.

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Transcellular passive diffusion is the commonest type of

absorption across membrane barriers

passive diffusion - a process that requires no molecular transport system or energy

source (random migration by individual solute molecules)

absorption rate for passive diffusion is determined by 3 major factors

passive diffusion cannot concentrate substances across membrane barrier (no pumping action)
bidirectional -- flow of molecules will follow the concentration gradient in either direction (in or out of tissue)

surface area through which diffusion is occurring (membrane lining of gut, lung, and skin)
concentration gradient [Cexternal] >> [Cinternal]
permeability of the substance through the membrane barrier

permeability is typically determined by each substances physicochemical properties

molecular weight
smaller molecules (MW < 500 daltons) are often able to migrate through biomembranes by passive diffusion
over 80% of effective drugs have a MW < 450 daltons
hydrophobicity
tendency of a substance to dissolve preferentially in fatty or oily biological media, but not in water
ionization
molecules that carry positively or negatively charged functional groups have ionic properties
charged ionic groups experience electrostatic interactions with ionic phospholipid membrane groups
polarity (hydrogen bonding)
molecules with uneven electrical charge distribution (polar compounds) form H-bonds with water

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Lipid sieve model of cell membrane

HYDRO
PHILE

The lipid sieve model helps

to explain how small
molecules that are lipophilic
can permeate through the
cellular phospholipid
membrane by passive
diffusion

EXTERIOR

HYDRO
PHILE

HYDRO
PHILE

polar heads
HYDRO
PHILE
LIPOPHILE

non-polar tails

Phospholipid
Bilayer

hydrophilic molecules cannot

permeate the membrane
unless there is a specific
paracellular transport channel
or membrane-associated
active transport pump.

INTERIOR

FACILITATED DIFFUSION
OR
ACTIVE TRANSPORT
HYDRO
PHILE

PASSIVE
DIFFUSION
LIPOPHILE

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Molecular dynamics computer simulation of

membrane diffusion during xenobiotic absorption

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Lipophilic and hydrophilic solubility

lipid solubility affects transcellular passive diffusion through the phospholipid

biomembranes

hydrophilic (water soluble)

lipophilic (fat and oil soluble)

ionic molecules carry one or more positive or negative charges

polar molecules carry partial positive or negative charges
phospholipid molecules on the membrane surface contain a zwitterionic charge distribution
negatively charged phosphate groups PO 4-positively charged choline groups N-[CH 3]4+
charged phospholipid groups will repel or bind ionic hydrophiles via electrostatic interactions
charged phospholipid groups will form hydrogen bonds (H-bonds) with uncharged hydrophiles that have
polar functional groups (esters, amides, etc.)
most hydrophiles cannot pass across membranes by transcelluar passive diffusion

electrically neutral molecules with no positive or negative charges

no electrostatic repulsion or H-bond attraction at the membrane surface
readily penetrate into and through the the non-polar interior of biomembranes
many small lipophiles can pass through biomembranes by transcellular passive diffusion
usually small lipophiles can be more readily absorbed than most small hydrophiles

lipophilicity factors are used to predict passive absorption of drugs and xenobiotics

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lipophilicity = hydrophobicity - [polarityH-bonding + ionic interactions]

calculated rate of absorption = 1/size (MW) x 1/lipophilicity (log K o/w)

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Partition coefficient is a quantitative measure of the

degree of lipophilicity of a given molecule

partition coefficient (Kp, Ko/w)

measure concentration of xenobiotic in 2-phase solvent mixture

oily non-aqueous phase solvent (octanol) and watery aqueous phase (H2O)
oil and water dont mix

Ko/w = conc (octanol) / conc (water)

measures relative degree of solubility in lipid (lipophilicity) and water (hydrophilicity)

Ko/w > 1 is lipophilic

Ko/w<1 is hydrophilic

Ko/w = 0 - 1 is amphiphilic (mixed)

log Ko/w often expressed in log10 units

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example:

Ko/w = 1000 --> log Ko/w = 3 (strongly lipophilic)

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Lipinskis rule of five for predicting xenobiotic

absorption by transcellular passive diffusion
Poor transcellular absorption and membrane
permeation is more likely when:

there are more than 5 H-bond donors in the molecular structure

(mainly OH and NH groups)

the molecular weight is over 500

the molecules log Ko/w is over 5
there are more than 10 H-bond acceptors in the molecular structure (mainly N
and O containing polar groups)

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Effect of lipophilicity on the absorption rate of 3 related

xenobiotic substances (barbiturate drugs)
ko/w

ko/w

ko/w

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Effect of partition coefficient on absorption rate

extremely
lipophilic
strongly
lipophilic

4-5

3
2

1
log Kp < 0 substances are
poorly absorbed due to
ionic interactions or H-bonding

hydrophilic

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<0

moderately
lipophilic

log Kp > 5 substances are poorly absorbed

due to membrane trapping
or lack of water solubility

0 - 0.9 mixed or
amphiphilic

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Absorption of large or non-permeable xenobiotic

molecules can occur via cellular endocytosis

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Absorption into brain of manganese (Mn2+) ions via

active transport channels and cellular endocytosis
TMI slide (illustrative purposes only)

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