Activetransport

1. Primary:ATPcoupled
2. Secondary:drivenbyiongradients

Passivetransport
1. Facilitateddiffusion:proteinmediatedtransport,notcoupledtoATPor
iongradients
2. Nonfacilitateddiffusionoflipophiliccompounds;nonionicdiffusion

Transportoftypes13ismediatedbytransportproteinsandissubstratespecific.Mostdrugs
dontfitthesubstratespecificitiesofanyofthesetransporters,sotheyhavetocrosscell
membranesbynonfacilitateddiffusion,whichoccursdirectlyacrossthelipidbilayerandis
independentofmembraneproteins.Theefficiencyofthismodeoftransportdependsmostlyon
thedrugsphysicochemicalproperties.

3.4.3 The polarity of drug molecules affects their rate of diffusion across
lipid bilayers

Theabilityofdrugmoleculestocrossalipidbilayerbypassivediffusioncorrelateswiththeir
abilitytopartitionintoandoutofthehydrophobicmembraneinterior.Afairlybutnotexcessively
hydrophobiccharacterismostconducivetotransport;verypolarmoleculeswillfailtoenterthe
membrane,whereasextremelyhydrophobicmoleculeswillenterreadilybutmayhaveahardtime
leavingitontheotherside.

The membrane permeability of drugs can be improved by removing polar

functional groups
Removalofchargedandpolarfunctionalgroupscanimprovetheabilityofadrugmoleculeto
crosslipidbilayers.Forexample,ephedrineandmetamphetamineareanaloguesofepinephrine
thathaveshedsomehydroxylgroups.Accordingly,theypenetratecellmembranesmorereadily
thantheparentcompound,whichpromotestheiruptakefromtheintestineandtheirdistributionto
thebrain.
However, the structural changes also cause a shift in the molecular mode of action. Like
epinephrine,bothephedrineandmetamphetaminestimulateadrenergicsynapses;however,while
epinephrine acts directly onadrenergic receptorsat the cell surface, ephedrine and
metamphetamineactprimarilyonintracellulartargets(seeslide6.14.3).Thisexampleillustrates
thatthescopeofstructuralalterationsforthesakeofimprovingpharmacokineticpropertiesis
limited.

Analternateapproachtoimprovingthemembranepenetrationofdrugmoleculesistoturnthem
intoprodrugs,inwhichpolarorionizablegroupsaremaskedwithhydrophobicresidues.After
uptake,thesehydrophobicgroupsmustbeabletoundergoenzymaticcleavageinordertorelease
theoriginaldrugmolecules.Mostcommonly,suchfunctionalgroupsareorganicesters.
Bacampicillinissucharesorptionester;itcontainsampicillin,whichafteruptakeintothe
intestinalepitheliumisreleasedbyesterases.Heroinismorphinewithtwoacetylgroupsaddedso
astomasktwopolarhydroxylgroups

Transfer through the lipid bilayer: passive diffusion

The transfer through a lipid bilayer is passive without requiring energy. The lipid bilayer membrane
constitutes a barrier:

impermeable to ions such as Na+, K+, Cl - , to polar molecules even non charged, i.e. nonionic, like glucose, and proteins.

permeable to non polar molecules (liposoluble or hydrophobic) of low or middle molecular

weight, and to molecules at the gas state and small molecules of low polarity.

Passive diffusion through a lipid bilayer

The passage through the membrane is directed from the most concentrated solution towards the least
concentrated solution until obtaining an equilibrium. The rate of transfer depends on the area S on the
membrane, on the concentrations C1 and C2 on both sides of the membrane and on a diffusion
constant K linked to the liposolubility and the size of the molecule (the smaller the molecule, the easier
the transfer).
V = K.S. (C2 - C1)
The liposoluble character of a molecule is determined by the measurement of its partition coefficient
between an aqueous solvent and an organic solvent like hexane. The liposoluble or non-polar molecules
accumulate in organic solvent and the polar molecules in water.
The polarity of a drug depends on its ionization and one distinguishes three categories:
1.

Permanently ionized molecules, whatever the pH, for example those with a quaternary
ammonium. These molecules, in theory, do not cross the lipid bilayer by passive diffusion.

2.

Neutral molecules, not ionized, whatever the pH. It is the case of the organic solvents which
cross the lipid bilayer easily. This rule has limitations, in particular concerning intestinal
absorption because it takes place from an aqueous medium. A very lipophilic molecule but
almost insoluble in water can be poorly absorbed. A low availability can be the consequence of
a low solubility in water.

3.

Molecules whose ionization depends on pH: in a neutral state, they cross the lipid bilayer , but
not in the ionized state. The acid drugs dissociate in a basic medium and the bases in an acidic
medium, to give ionized molecules. The pKa of an acid is the pH at which it is 50% dissociated.
For example, an acid drug R-COOH partially dissociated in R-COO - + H + , neutral form RCOOH crosses membranes but not the ionized form R-COO - .

Consequently, when two compartments at different pH are separated by a membrane, the equilibrium of
the concentrations of a drug R-COOH on both sides of the membrane will be moved and drug R-COOH
will accumulate in the basic compartment where it dissociates.

Transmembrane transfer of an acid drug according to pH

Binding of the drugs to plasma proteins, in particular to albumin, which can go from 0% to 99% modifies
their transfer through the membranes. In the free state, i.e. unbound to plasma proteins, the liposoluble
drugs, if there is a gradient of favorable concentration, cross the lipid membrane, whereas the fraction
bound to plasma or tissue proteins does not cross it.

Transmembrane transfer of a drug according to its protein binding

Albumin is a protein with a molecular weight of 68000 daltons, present in plasma at a concentration
approximately 50 g/L. It is synthesized by the liver and degraded primarily by the vascular endothelium.
Its plasma half-life is approximately 20 days. In addition to its role in oncotic pressure, it binds various
endogenous molecules and drugs. There are on a molecule of albumin six different types of sites with
different affinities for drugs.
The equilibrium for a drug between the free fraction, D, and the bound fraction, DP, is reversible and
non-static in the body because the blood circulation causes dynamic changes.

Blood cells, mainly red cells, can play a role comparable to that of albumin for drugs that they bind or
incorporate.

Transfer through membrane protein structures

The transfer through the protein structures of membranes is carried out by active transport, i.e. using
energy provided by cellular metabolism.
Direct active transport by pumps
The active transport requires energy, generally provided by ATP hydrolysis. The Na+/K+-ATPASE pump,
Mg2 + dependant, uses the energy of ATP to polarise the cell by extruding three Na+ ions and
incorporating two K+ ions, which creates a potential difference between the intracellular medium and the
extracellular medium. There are other pumps: a Ca2+-ATPASE pump localized on the level of the
cytoplasmic membrane and at the level of the endoplasmic reticulum and an H+ /K+-ATPASE pump.
A particular membrane protein, P-glycoprotein, or P170 because its molecular weight is 170 Kd, uses,
like the Na+/K+-ATPASE pump , the energy supplied by the hydrolysis of ATP to drive certain drugs out
of the cell. This glycoprotein, also called Multi drug transporter, protects the cell against xenobiotics but,
as it is expressed in the majority of cancerous cells, it also expel antineoplastic drugs out of cancerous
cells, thus explaining their resistance to chemotherapy which is called MDR Multi drug resistance.
Drugs such as verapamil or quinidine inhibit P-glycoprotein but have other properties making their use
as inhibitors of P-glycoprotein difficult. P-glycoprotein present in the blood-brain barrier reduces
penetration of certain drugs into the brain. It can also reduce their digestive absorption.
Various microorganisms have in their plasma membrane, in addition to P-glycoprotein, protondependant pumps which drive toxic molecules out of their cytoplasm.
Indirect active transport
The energy necessary for facilitated diffusion, or secondary active transport, is supplied by the ion
gradients on both sides of the membrane. It is not carried out through the lipid bilayer but through the
protein structures.
It relates to only a small number of compounds:
1.

molecules implicated in metabolism, like glucose, amino acids, certain transmitters and drugs
of close chemical structure. The energy necessary for their transport can be supplied by the
sodium gradient.
The kinetics of transfer is of Michaelis Menten type with maximum rate and possibility of
competition between molecules. When sodium and the substrate cross the membrane in the
same direction, the transport is called symport and antiport when they cross in opposite
directions.

2.

peptides formed of 2 or 3 acids amino are partially absorbed by the digestive tract and
reabsorbed in the nephron by specific carriers using the H+ gradient as source of energy.
These carriers are involved in the digestive absorption of drugs with a chemical peptidic-like
structure such as beta-lactamines. A polypeptide such as insulin can cross the blood-brain
barrier but the responsible mechanisms are poorly understood.

3.

ions like Na+, K+, Ca2+, Cl _ , whose membrane transporters are channels and exchangers:
o

channels whose opening and closure depend either on the intra/extracellular potential
difference (opening during depolarization), or on the presence of receptors which can
be activated or inhibited by various transmitters

exchangers, Na+/Ca2+ for example.

Transport by exocytosis and endocytosis

Exocytosis consists of the exit out of the cell of molecules contained in vesicles which fuse with the
plasma membrane and release their contents outside. It is the mode of release of transmitters.
Endocytosis consists of the absorption by a cell of an extracellular molecule. After its inclusion in a
vesicle formed by an invagination of the plasma membrane, the molecule penetrates into the cytoplasm.
It is, for example, the process used by hepatocytes to take up lipoproteins and transferrin,.
Oligonucleotides of 10 to 20 units, such as those used in antisense therapy (oligonucleotides
complementary to the mRNA on which they bind) penetrate into cells by various mechanisms but
primarily by endocytosis. In fact, an oligonucleotide penetrates better into cells that an isolated
nucleotide.

nstead, the electrochemical potential|electrochemical potential difference created by

pumping ions out of the cell is used.