BIOPHARMACEUTICS

ABSORPTION
SYSTEMIC ABSORPTION IS DEPENDENT ON:
•The anatomy and physiology of the drug absorption
site
•The physicochemical properties of the drug
•The nature of the drug product/formulation factors
 Physiological Factors Affecting Oral
Absorption
Objective:
To understand the physiological factors which affect the oral
absorption of drug products
Physiological Factors:
A. Membrane physiology:
– Considering the structure of membranes
– Transport processes
B. Gasstrointestinal physiology:
– Characteristics of gastrointestinal physiology
– Gastric motility and emptying
– Influence of food
– Other factors
 Diagram Scheme of ADME
processes

DISINTEGRATION DISSOLUTION ABSORPTION

PHARMACEUTICAL PHASE Protein Binding PHARMACOKINETIC PHASE

TISSUE

DRUG IN
SYST.CIRCULATION: THERAPY
FREE ⇄ BIND DISTRIBUTION RECEPTOR PHARMACOLOGI
AL EFFECT

METABOLISM TOXIC

ELIMINATION PHARMACODYNAMIC PHASE

Excretion
• The ultimate goal of drug absorption is to
have the drug reach the site of action in a
concentration which produces a
pharmacological effect. No matter how
the drug is given (other than I.V.) it must
pass through a number of biological
membranes before it reaches the site of
action.
 A. Membrane physiology
1. Membrane structure
• In 1900 Overton performed some simple but
classic experiments related to membrane
structure. By measuring the permeability of
various types of compounds across the
membranes of a frog muscle he found that
lipid molecules could readily cross this
membrane, larger lipid insoluble molecules
couldn't and small polar compounds could
slowly cross the membrane.
Diagram XI-2, the Davson-Danielli
Model
Model of the plasma membrane including proteins and carbohydrates as
well as lipids. Integral proteins are embedded in the lipid bilayer;
peripheral proteins are merely associated with the membrane surface.
Carbohydrate attached to proteins: glycoproteins, to lipid: glycolipids.
These proteins provide a pathway for the selective transfer of certain
polar molecules and charged ions through the lipid barrier
Fluid Mosaic Model by Singer and Nicholson
• These results suggest that the biologic
membrane is mainly lipid in nature but
contains small aqueous channels or pores.
Other experiments involving surface
tension measurements have suggested that
there is also a layer of protein on the
membrane. These results and others have
been incorporated into a general model for
the biological membrane. This is the
Davson-Danielli model.
• The membrane then acts as a lipid barrier
with small holes throughout.
 Nature of the drug transport in
the body
• Transcellular Transport
• Paracellular Transport
• Intestinal Epithelial cell Transport
 Diagram XI-3, Simplified
Model of Membrane

Transcellular

Paracellular
This is the general structure. Membranes in different parts of the body
have somewhat different characteristics which influence drug action and
distribution. In particular, pore size and pore distribution is not uniform
between different parts of the body.
Examples of some membrane types.
• Blood-brain barrier. The membranes between
the blood and brain have effectively no pores. This
will prevent many polar materials (often toxic
materials) from entering the brain. However,
smaller lipid materials or lipid soluble materials,
such as diethyl ether, halothane, can easily enter
the brain. These compounds are used as general
anesthetics.
• Renal tubules. In the kidney there are a number
of regions important for drug elimination. In the
tubules drugs may be reabsorbed. However,
because the membranes are relatively non-porous,
only lipid compounds or non-ionized species
(dependent of pH and pKa) are reabsorbed
• Blood capillaries and renal glomerular
membranes. These membranes are quite porous
allowing non-polar and polar molecules (up to a
fairly large size, just below that of albumin, M.Wt
69,000) to pass through. This is especially useful in
the kidney since it allows excretion of polar (drug
and waste compounds) substances.
• Placenta Membranes
• Testis Membranes
 Transcellular Transport/pathway
• Passive Diffusion
• Carrier Mediated/Facilitated Transport or
Diffusion : Active transport, Facilitated
transport, Carrier-Mediated Intestinal
transport
• Vesicular Transport. Endocytosis/Exocytosis :
pinocytosis, phagocytosis, receptor-mediated
endocytosis, transcytosis
• Pore (Convective) Transport
• Ion pair transport
 Transport across the membranes
(Transcellular Transport)
a)Passive Diffusion with a Concentration Gradient

• CGI

CP

h
• Most drugs cross biologic membranes by
passive diffusion. Diffusion occurs when
the drug concentration on one side of the
membrane is higher than that on the other
side.
• Drug diffuses across the membrane in
an attempt to equalize the drug
concentration on both sides of the
membrane.
• If the drug partitions into the lipid
membrane, a concentration gradient
can be established.
 Equation 1: Rate of Diffusion
Fick’s law of Diffusion
• The rate of transport of drug across the
membrane can be described by Fick's
first law of diffusion:
dQ DAK
• Rate of diffusion = = ( CGI − CP )
dt h
dQ/dt = rate of diffusion; D = diffusion coefficient; K =
lipid water patition coefficient of drug; A = surface area
of membrane; h = membrane thickness; CGI-CP =
difference between the concentration of drug in GIT and
in the plasma
b)Carrier Mediated /Facilitated Transport or Diffusion
1.Active transport. The body has a number of specialized
mechanisms for transporting particular compounds; for
example, glucose and amino acids. Sometimes drugs can
participate in this process; e.g. 5-fluorouracil. Active
transport requires a carrier molecule and a form of energy
• Energy consuming process, ATP hydrolysis or
transmembraneous sodium gradient and/or electrical
potential
• Carrier/transporter mediated, the process can be saturated
• against a concentration gradient across cell membrane
• competitive inhibition is possible : metabolic inhibitors or
substrate analogues
• Temperature dependence
• Important role in the intestinal, renal and biliary excretion
of many drugs
Four types of membrane transport proteins couple the energy-
releasing hydrolysis of ATP with the energy-requiring
transport of substances against their concentration gradient

• P-glycoprotein is one of the transmembrane

protein acts as a carrier-mediated intestinal
transporter identified in the intestine
• Pgp appears to reduce apparent intestinal
epithelial cell permeability from lumen to
blood for various lipophilic or cytotoxic
drugs
 P-class Pumps
• P-class pumps are composed of two different polypeptides,
α and β, and become phosphorylated as part of the transport
cycle. The sequence around the phosphorylated residue,
located in the larger α subunits, is homologous among
different pumps
• In P-class pumps, phosphorylation of the α subunits and a
change in conformational states are essential for coupled
transport of H+, Na+, K+, or Ca2+ ions
• The P-class Na+/K+ ATPase pumps three Na+ ions out of
and two K+ ions into the cell per ATP hydrolyzed. A
homolog, the Ca2+ ATPase, pumps two Ca2+ ions out of
the cell or, in muscle, into the sarcoplasmic reticulum per
ATP hydrolyzed. The combined action of these pumps in
animal creates an intracellular ion milieu oh high K+.ow
Na+ very different from the extracellular fluid milieu of
high Na+, high Ca2+, and low K+
 F-class and V-class pumps
• F-class and V-class pumps do not form
phosphoprotein intermediates. Their structures are
similar and contain similar proteins but none of their
subunits are related to those of P-class pumps
• In the multisubunit V-and F-class ATPases, which
pumps protons exclusively, a phosphorylated protein
is not an intermediate in transport
• A V-class H+ pump in animal lysosomal and
endosomal membranes and plant vacuola membranes
is responsible for maintaining a lower pH inside the
organelles than in the surrounding cytosol
 ABC superfamily
( ATP Binding Catalytic domains or Cassete)
• All members of the large ABC superfamily of proteins
contain four domains: two transmembrane (T) domains and
two cytosolic ATP-binding (A) domains that couple ATP
hydrolysis to solute movement. These core domains are
present as separate subunits in some ABC proteins, but are
fused into a single polypeptide in other ABS proteins
• All members of the large and diverse ABC superfamily of
transport proteins contain 4 core domains: two
transmembrane domains, which form a pathway for solute
movement and determine substrate specificity, and two
cytosolic ATP-binding domains
• The ABC superfamily includes bacterial amino acid and
sugar permeases; the mammalian MDR1 protein, which
export a wide array of drug from cells; CTFR protein, a Cl
channel that is defective in cystic fibrosis
Carrier Mediated Transport Process
2.Facilitated Diffusion
• A drug carrier is required but no energy is
necessary. e.g. vitamin B12 transport
• saturable if not enough carrier, subjected to inhibition
by competitive inhibitor
• no transport against a concentration gradient, downhill
but faster
• Requires concentration gradient for its driving force, as
does passive diffusion
• Much faster rate than would be anticipated based on the
molecular size and polarity of the molecule
• Minor role in drug absorption
 3.Carrier-Mediated Intestinal Transport
• Various carrier-mediated systems (transporters) are
present at the intestinal brush border and basolateral
membrane for the absorption of specific ions and
nutrients essential for the body. Many drug are
absorbed by these carriers because of the structural
similarity to natural substrates (see Table)
• Other carrier-mediated intestinal transporter:
amino acid transporter, oligopeptide transporter,
phosphate transporter, bile acid transporter,
glucose transporter, monocarboxylic acid
transporter
• Many oral cephalosporins are absorbed through
the amino acid transporter. Cefazolin, a
parenteral-only cephalosporin, cannot be
absorbed through this mechanism, is not
available orally
 Intestine Transporters and Examples of Drug Transported
Transporter Examples
Amino acid Gabapentin p-cycloserine
Methyldopa Baclofen
L-dopa
Oligopeptide Cefadroxyl Cephradine
Cefixime Ceftibuten
Cephalexin Captopril
Lisinopril Thrombin inhibitor
Phosphate Fostomycin Foscarnet

Bile acid S3744

Glucose P-nitrophenyl-β-D-glucopyranoside
P-glycoprotein efflux Etoposide Vinblastine
Cyclosporin A
Monocarboxylic acid Salicylic acid Benzoic acid
Pravastatin
 Figure 13-1 from your book p.374
• Summary of intestinal epithelial transporters.
Transporters shown by square and oval shapes
demonstrate active and facilitated transported,
respectively
• Name of cloned transporters are shown with
square or oval shapes
• Active transporters: arrows in same direction
represent symport of substance and the driving
force
• Arrows going in the reverse direction mean the
antiport
 Efflux of drugs from the intestine
• Counter transport efflux proteins that expel
specific drugs back into the lumen of GIT after
they have been absorbed
• Example: P-glycoproteins
• Requires energy
• Against a concentration gradient
• Competitively inhibited by structural analogues
or metabolism inhibitors
• Saturable process
 Transport across Cell Membranes
Active Transport by ATP-Powered Pumps
Figure 15-17. Possible mechanisms of action of the MDR1
protein. (a) The flippase model proposes that a lipid-
soluble molecule first dissolves in the cytosolic-facing
leaflet of the plasma membrane ( 1 ) and then diffuses in the
membrane until binding to a site on the MDR1 protein that
is within the bilayer ( 2 ). Powered by ATP hydrolysis, the
substrate molecule flips into the exoplasmic leaflet ( 3 ),
from which it can move directly into the aqueous phase on
the outside of the cell ( 4 ). (b) According to the pump
model, MDR1 has a single multisubstrate binding site and
transports molecules by a mechanism similar to that of other
ATP-powered pumps. [Adapted from G. Ferro-Luzzi Ames
and H. Legar, 1992, FASEB J. 6:2660; N. Nelson, 1992,
Curr. Opin. Cell Biol. 4:654; C. F. Higgins and M. M.
Gottesman, 1992, Trends Biochem. Sci. 17:18; and C. F.
Higgins, 1995, Cell 82:693.]
 Physiological Role of P-Glycoprotein
• P-glycoprotein is found in high levels at apical
surface of cells typically associated with
transport of: biliary canalicular membrane,
brush border of renal proximal tubules, apical
surface of intestinal mucosal cells, endothelial
cells of brain and testis
• It has been proposed that the normal
physiological role of P-glycoprotein is one of
detoxification through active secretion of
xenobiotics
 Role of P-glycoprotein in cancer
• Approximately 50% of human cancers express P-
glycoprotein at levels sufficient to confer MDR
• Cancers which acquire expression of P-
glycoprotein following treatment of the patient
include leukemias, myeloma, lymphomas, breast,
ovarian cancer; preliminary results with P-gp
inhibitors suggest improved response to
chemotherapy in some of these patients
• Cancers which express P-gp at time of diagnosis
include colon, kidney, pancreas, liver; these do not
respond to P-gp inhibitors alone and have other
mechanisms of resistance
 P-Glycoprotein and Multi Drug
Resistance

• Multi Drug Resistance (MDR) is the phenomenon

whereby cancer cells develop resistance to
cytotoxic drugs
• MDR is a result of over expression of P-glyco-
protein: - MDR1 in human;
- mdr1 and pgp1 in rodents
• P-glycoprotein utilizes ATP hydrolysis to pump
cytotoxic drugs out of cells
P-glycoprotein as a transmembrane drug efflux pump

• The Multi Drug Resistance gene MDR1, which encodes the

cell-surface molecule P-glycoprotein (PGP) can confer
resistance to a wide variety of drugs. PGP transport drugs
out of the cell, which is a process that requires the presence
of two ATP binding domains.These domains are a defining
characteristic of this family of ATP Binding Cassete (ABC)
transporters.
• The exact mechanism of drug efflux is not well understood,
but might involve either direct transport out of the
cytoplasm or redistribution of the drug as it transverses the
plasma membrane. Some cytotoxic drugs that are known
substrates for PGP include etoposide, daunomycin, taxol,
vinblastine and doxorubicin. PGP is modified by sugar
moieties on the external surface of the protein
c) Vesicular Transport/Endocytosis&Exocytosis:
The processes of moving specific macro-molecules
into and out of cells, respectively
Pinocytosis&Phagocytosis:
• Engulfment particles or dissolved materials by the
cell
• For example Vitamin A, D, E, and K, Sabin polio
vaccine and various large proteins.
• Receptor-mediated endocytosis
• Transcytosis
d)Pore (Convective) Transport
e) Ion pair transport
• For example quaternary ammonium compounds
 Pore (Convective) Transport
• Very small molecules such as urea, water and
sugars, are able to cross cell membranes rapidly, as
if the membrane contained channels or pores. The
model of drug permeation through aqueous pores is
used to explain renal excretion of drugs and uptake
of drugs into the liver
• A certain type of protein called a transport protein
may form an open channel across the lipid
membrane of the cell. Small molecules including
drugs move through the channel by diffusion more
rapidly than at other parts of the membrane
 Ion Pair Transport
• Strong electrolytes drugs are highly ionized or charged
molecules, such as quaternary nitrogen compounds with
extreme pKa values – maintain their charge at all
physiologic pH values and penetrate membranes poorly
• When the ionized drug is linked up with an oppositely
charged ion, an ion pair is formed with neutral charge –
diffuses more easily across the membrane : propranolol (a
basic drug) paired with oleic acid; quinine paired with
hexylsalicylate; complexation of Amphotericin B and
DSPG (DiSteroylPhosphatidylGlycerol)
• Ion pairing may transiently alter distribution, reduce high
plasma free drug concentration, and reduce renal toxicity
 Paracellular Transport/pathway
• Water and small hydrophylic molecules pass through
numerous aqueous pores
• Transport of material across aqueous pores between the
cells
• The cells are joined together via closely fitting tight
junctions on their apical side
• Generally, absorptive epithelia tend to be leakier than other
epithelia, decreases in importance down the length of the
GIT (decreases in number and size of pores)
• Important for the transport of ions, sugars, amino acids and
peptides at concentration above the capacity of their carriers
• Small, hydrophilic and charged drugs
• Molecular weight cut-off: 200 Da
• Convective (solvent drag) and diffusive component
 The Mechanism of Paracellular
Transport
• Filtration
• Bulk flow
 B. Gastrointestinal
physiology
• Look at the GI Tract file
• Conclusion of factors affecting GIT absorption
rate:
1. Coefficient Partition between lipid-water
2. Local blood flow
3. Intestine surface area
4. Gastric emptying time
5. Gastrointestinal motility
6. Intestinal motility
7. Food
8. Formulation factors
 First-pass metabolism
• Drugs may be absorbed well, but still fail to
reach the systemic circulation
• All blood from the gut (except mouth and
lower rectal) passes through the portal
system to the liver
• Many drugs are extracted and metabolized
on their first-pass – may inactivate the
drugs
• The alternative route: bucally (glyceryl
trinitrate)
 Influence of dietary components on the
gastrointestinal metabolism and transport of
drugs
• Ingestion of meal ~ physiologic changes
(gastric pH, gastric emptying, hepatic blood
flow, etc) that significantly alter the rate and
extent of drug absorption
• Components of food ~ alter drug absorption
through alteration in drug solubility.
Nutritional status ~ variability in the
pharmacokinetic of certain drugs
• Grape fruit juice can increase the BA of certain
drugs, by reducing presystemic intestinal
metabolism, led to renewed interest in food-drug
interactions
• Effects of grapefruit flavonoid, naringin, and
furanocoumarin, 6’-7’-dihydrocybergamottin, on
the activity of CYP3A4. The possibility of grape
fruit juice might affect drug absorption via
interaction with intestinal P-glycoprotein (P-gp) is
being explored
• The use of herbal extracts, phytopharmaceutical
raise: cause changes in pharmacokinetics of
conventional drugs?
 Absorpsi non Oral
• Nasal Drug Delivery
• Inhalation Drug Delivery
• Topical and Transdermal Drug Delivery
NASAL
• Nasal mucose ≅ sublingual mucose : good
absorption
• Systemic - richly supplied with blood vessel:
α-adrenergic for infants
• Local Decongestant : rhinitis
• Diabetes incipidus : Desmopressin
• Carcinoma prostat : Gonadi Liberin analog
( oligopeptide, damage in GIT)
 Inhalation Drug Delivery
• Local/systemic
• Surface area 70 m2
• Bronchodilator
• Small particle droplet size
 Topical Delivery
• Usually : local,
• now with Transdermal Drug Delivery:
systemic – patch.
• Advantages: continuous release of drug
over a period of time, low presystemic
clearance, good patient compliance
• Scopolamin, Nitroglycerin, Estradiol, HRT
• Sometimes : local iritation
 Skin Absorption
• Transepidermal
• Absorption barrier: non vascular stratum corneum
(less water content: ± 10%)
• Absorption barrier : reservoir
• Lipophylic compound with small hydrophylicity :
increase per cutan absorption
• Hydrophobic compound : fat, oil, showed low per
cutan absorption because stratum corneum has less
lipid
• Skin penetration for lipid-insoluble drug
happened through hair folicle, sweat glands,
sebaceae glands
 Factors enhancing Skin Penetration
• Increasing skin temperature
• Using hyperemic stimuli : DMSO
• Increasing water content/hydration by compound
like urea
• Irritated tissue
• Damaging stratum corneum mechanically,
chemically, heat, burnt and wound
• Absorption rate depends on age, infants good
absorption through skin because has not yet
developed. Be careful to give corticosteroid cream