ABSORPTION

Absorption is the process of movement of unchanged drug molecule from the site of
administration to systemic circulation. Cell membrane act as a barrier for drug movement .It consist of
double layer of amphiphilic phospholipids, the membrane structure looks like Mayonnaise sandwich
model .

Cell membrane having thickness of 25 A˚ constitute to layers of proteins and 4 to 10 A˚pore size between
cells of proteins .

Lipid layer occupies middle position with phospholipids polar head and towards protein membrane ,non
polar tails towards lipid cells .

MECHANISM OF DRUG ABSORPTION :

Transcellular pathway ( across cell )

Paracellular pathway ( between cell gap )

Vesicular pathway ( engulf ) .

TRANSCELLULAR PATHWAY :

Most of the drugs follow this type

Based on energy consumption ;categorized into ;

(i) Active (ii) passive

ACTIVE TRANSPORT :

The help of energy molecules ( ATP ) drug molecules are transported from lower concentration to
higher concentration ( uphill movement ) .

The movement may be primary or secondary

In primary movement only ,drug molecules move while secondary movement along with nutrient drug
molecules .( ex: absorption of glucose ) .

PRIMARY :
Primary transport may be with ions or proteins , ion transport (ex: organic anion absorb drug like statins .

Diphenhydramine ( anti histamine ) transport through cations

p- glycoprotein (PGP ) called as ABC transport that is ATP binding cassettes.

Through this efflux mechanism ,hydrophobic drugs are diffused into brain ,this mechanism is called
exsorption .

SECONDARY :

Secondary transport depends on direction of movement ,they may be symport ( co- transport –same
direction ) . ex: beta lactum antibiotics are transported along with nutritient in same direction .

Antiport or counter transport ,nutrient or drug move in opposite direction ( ex: sodium absorption with
potassium depletion in renal .

PASSIVE TRANSPORT :

Without requiring much energy molecule from higher to lower concentration is called passive diffusion
(down hill movement )

This transport classified into 3 categories;

 Passive diffusion (Fick diffusion )

 Facilitate diffusion

 Core transport.

FICK DIFFUSION:

Through a semi- permeable membrane molecule diffuse from higher to lower concentration without
energy requirement is called FICK’s DIFFUSION .

The diffusion rate mainly depends upon concentration gradient ( concentration difference between both
side of membrane ).

Amount of molecule diffused ( flux) = dq/dt = DA Ko/w(C GIT – Cp) / h

Concentration gradient = CGIT – Cp .

D= diffusion coefficient of drug

A= area of membrane

Ko/w = partition coefficient of drug

h = thickness of membrane

According to flux ,rate of diffusion is directly proportional to surface area of membrane .

Small intestine maximum surface area, hence maximum absorption takes place .

The rate of diffusion is indirectly propotional to thickness of membrane .

Gastric membrane very thick when compared to intestine hence absorption lowers .

Higher partition coefficient increase rate of diffusion .

For continuous absorption, concentration gradient should be positive that is concentration of drug in GIT
> concentration of drug in plasma.

To maintain this (+) concentration gradient either periodic addition of drug in GIT fluid or replacement
of dissolution medium in invitro studies are done .this condition called SINK condition .

CORE TRANSPORT:

Solvent molecules easily filtered through 4 channels without requiring energy.

Urea, water, sucrose follows this absorption pathway .’

This movement otherwise called as solvent drag, convective transport bulk flow .

The hepatic circulation, low molecular weight substances follow this transport .

FACILITATED DIFFUSION

With the help of enzyme proteins or ions, molecules are transported without energy. This diffusion is
called as facilitated or carrier mediated transport.

Cationic drugs ex; propranolol hydrochloride transported through endogenous anion by forming a
complex

Enzyme proteins act as transport carrier for the false nutrients like vitamins and chemotherapeutic drugs
like 5 flurouracil or 5 bromo uracil.
Enzymes are susceptible for saturation and follows non linear Michaels Menton kinetics

Entry of glucose into RBC, intestinal absorption of vitamin B 12 are facilitated by biochemical intrinsic
factors.

PARACELLUAR PATHWAY S:

When the molecules move between the cell is called paracellular or intra cellular transport.

Aqueous pores present in GIT easily absorb cardiac glycosides.

The openings may be temporary or permanent. These absorption are also called as persorption .

VESICULAR TRANSPORT:

It is a minor transport mechanism in which engulfment of drug molecules takes place in the form of
vesicles.

This process is called endocytosis or vesicular transport, it may be phagocytosis ( cell eating ) i.e.solid
molecules are taken up by cell membrane .

Pinocytosis( cell drinking) for liquid molecules,fat soluble vitamin A,D,E,K follows this type of
absorption .

Oral polio vaccine is absorbed through phagocytosis.

After dissolution pre-uptake phase molecules taken up by GIT membrane and transported to systemic
circulation either to hepatic or lymphatic .

High molecular weight liphophilic drugs follow lymphatic pathway whereas lower molecular weight
follows hepatic pathway or circulation .Some drugs before reaching liver get eliminatd that is called pre-
systemic elimination or first pass metabolism .

FACTORS AFFECTING GI ABSORPTION OF DRUGS :

There are 2 factors mainly affecting GI absorption .

(i) Drug & dosage form related factors ( pharmaceutical factors )

(ii) Physiological and pathophysiological factors .

Drug and dose related factors:

 Physiochemical characters of drugs significantly affect rate of absorption, mainly the following
characters affect absorption;

a. Solubility and dissolution

b. Particle size and surface area
c. Polymorphic nature of the drug
d. Molecular water
e. Salt and ester form of drugs.
f. Stereo chemical nature of drug
g. Lipophilic nature of drug with its ionization.

SOLUBILITY AND DISSOLUTION:

Amount of solid dissolved in a solvent at particular temperature ,pressure and pH is called absolute
solubility .

According to BCS system drugs are classified into 4 classes;

Class I – high soluble, high permeable (HS, HP) ex: diclofenac sodium

Class 2- low soluble, high permeable (LS, HP) ex: nimesulide

Class 3: high soluble low permeable (HS, LP) ex: ranitidine HCl

Class 4: low soluble low permeable (LS, LP) ex: proteins and peptides .

For GIT absorption, first 3 classes of drugs are converted as formulations.

Class 1 drug absorption is directly proportional to their solubility poor soluble drugs are converted into
salt form to increase the solubility.

Solublizers like surfactant or co-solvent are added in class 2 drugs to increase the solubility.

Dissolution theories are to explain dissolution, diffusion layer model theory (Philip theory) .

NOYE’S WHITNEY EQUATION :

Rate of dissolution dc/dt = DA K w/o ( Cs – Cb) / Vh

D- Diffusion coefficient of drug

A – Area of the membrane on diffused layer

Kw/o - partition coefficient between GI fluid and plasma

Cs - concentration of drug in stagnant layer

C b – concentration of drug in bulk layer

V – Volume of solvent

h - Thickness of GIT membrane.

According to the dissolution equation , reducing the particle size increase the dissolution; minimizing the
fluid intake with drug formulation minimize the dissolution ,thickness of membrane formulation
reduces the dissolution the concentration gradient ( Cs –Cb) should be positive (driving force for
dissolution) .periodical withdrawal of dissolution medium and replacement with buffers maintains sink
condition

During dissolution, solid disintegrated, then de-aggregated, the deaggregated particles get dissolved and
diffused.

Dissolution is the rate limiting step for absorption.

POLYMORPHIC NATURE OF DRUG:

When drug exist more than one physical form is called polymorphic form.

Stable polymorphs are easily interchangeable

Chloramphenicol palmitate available in forms where A is active & B is inactive

Sulfonamide, barbiturates show different type of polymorphic nature.

Proper isolation of polymorph is required for better absorption

MOLECULAR WATER:

Solvent water gets entrapped into molecular structure of drug and become solid

In some cases, it encircles the drug moiety and acts as hydrates

Hydrated solids are called pseudo polymorphs presence of water molecule lowers the absorption
SALT AND ESTER FORM OF THE DRUG:

Acidic and basic salt form of drug easily gets ionized and dissolved where as ester forms of the drug
behave like prodrug reduces the dissolution.

Eg. Erythromycin HCl (salt) used for immediate release, erythromycin besylate (ester) used for SR
tablets.

STEREOCHEMICAL NATURE:

Most of drug available as racemic mixture

Most of levoform produce higher therapeutic activity than dextro form

Eg. Levocetrizine highly active than cetrizine

LIPOPHILICITY AND IONIZATION OF DRUG (pH PARTITION THEORY)

This theory says GIT membrane is a lipid membrane and unionized drug having lipid nature easily diffuse
through this membrane. But ionisation mainly depends upon the pH if absorption site.

According to Henderson Hasselberg equation

Relation between pKa and pH

Weak acid pH= pKa + log [ionized]

[unionized]

Weak base pH= pka + log [unionized]

[ionized]

Most of drugs are either weak acid or weak base so they do not get 100% ionisation.

When ionisation and unionization become equal pKa = pH (When 50% ionized and 50% unionized
pH=pKa)

According to this theory, Strong acid and Strong base are rejected from absorption.

Very weak acid like phenobarbital is unionized at all pH and easily get absorbed.
Moderate weak acid like aspirin, unionized in gastric pH and absorbed, whereas ionized in intestine
shows poor absorption.

Very weak base like theophylline, unionized at all pH, equally absorbed in stomach and intestine.

Moderate weak base like codeine ionized at gastric pH but unionized at intestinal pH and absorbed.

Strong acid like disodium bromoglycolate and Strong Base like guanethidine shows poor absorption at
GIT because of ionization at all pH.

HLB (Hydrophilic Lipophilic Balance) play major role in partition coefficient.

Hydrophobic drugs easily diffuse through lipid membrane whereas water soluble drug shows poor
absorption.

Modifications/ Limitations of Theory:

This theory is lacking in explaining the following:

a) Ionized drug absorbed through ion via transport, but this theory states ionized drug will not get
absorbed.
b) This theory states about membrane nature (lipid) but stomach and intestine shows different levels
of absorption because of surface area and residence time (intestine large surface area and
residence time >6 hours, stomach limited surface area with 2-3 hours residence time).
c) Presence of virtual membrane (stagnant layer) acts as another barrier for absorption.
d) Presence of peristaltic movement or stirring changes the dissolution to significant level.

LIPINSKI derived a formula called Rule of 5:

According to it:
Molecular weight of drug <500 easily gets absorbed.
If lipophillicity, logP<5 easily get absorbed.
If no. of H bond donors <5 easily gets absorbed.

DOSAGE FORM RELATED FACTORS:

After disintegration, oral solid formulation gets dissolved and absorbed, delay released
formulations like enteric-coated tablets are intact in gastric fluid and easily dissolved in alkaline
pH of intestine.
o Polymers mainly used as excipients to modify release pattern, aqueous polymers increase
the dissolution rate while insoluble polymers slow down the dissolution rate.
o Granulation methods and tablets and packing of granules and capsules affect absorption
significantly.
o Wet granulation facilitates disintegration, dry granulation retards disintegration. Higher
compression force increase hardness of tablet and decreases disintegration.
o Tightly packed granules and capsules slow down the absorption.
o Liquide orals containing aqueous vehicle (syrup) makes faster absorption while glycerine
added liquid extract slow down the absorption.
o Diluents are generally added to increase the bulkiness,but sometimes it makes physical
incompatibility by forming complexes.
o Eg: Calcium phosphate forms insoluble complex with tetracycline.
o Binders are added to make cohesive mass, insoluble binders like ethyl cellulose reduces
the absorption rate.
o Disintegrants are added to facilitate disintegration, but higher concentration of
disintegrant slow down release of drugs Eg: microcrystalline cellulose at higher
concentration, slow down drug release.
o Lubricants and glidants are added to increase the flow property of granules.
o Eg: magnesium stearate as lubricants slow down absorption but talc as glidant increases
absorption.
o Suspending agents added in liquid-orals increases the viscosity and reduces the
absorption rate.
o Surfactant reduces the surface tension but increases the absorption.
o The order of absorption for oral formulation:
Solution > emulsion > suspension > capsules > tablets > coated tablets > enteric coated
tablet > sustained release tablet(SR).
o Suspension are semisolids, easily get disintegrated and absorbed.
o Capsules containing gelatin easily get hydrated and dissolved.
o Enteric coated and sustained release tablets slow down disintegration, dissolution and
absorption.
PATIENTS RELATED BIOLOGICAL FACTORS:
Physiological factors:
Length of GIT is 415 cm, major component of GIT are stomach, small intestine, large
intestine, ileum.
Comparative pH, surface area, blood flow and transit time are as follows:

GI parts pH Surface area Blood flow Transit time

Stomach 1.2 0.2 sq. meter 0.15L/ min 2-3 hrs
Small intestine 6.7 200 sq. meter 1L/ min 3-6 hrs
Large intestine 7.8 0.15 sq. meter 0.02L/ min 6-12 hrs

-Small intestine have larger surface area due to folding (kercking).

-Each kercking have villi and microvilli (folding in foldings)
-So, surface area increase upto 30 folds
-This increment in surface area plays a major role in absorption than stomach.
-Blood flow 1L/ min in small intestine, further increases the absorption.
-Gastric emptying time is within 2-3 hrs whereas in intestines, it is 18 hrs.
-Maximum residence time in intestine favors absorption of basic drugs.
-Azopolymers disintegrate in the presence of anaerobic bacteria are used for colon cancer
of drugs
Ex: 5- aminosalicylic acid

PATHOPHYSIOLOGICAL FACTOR:
-Pediatric and geriatric patients have different surface area and GIT have varying absorption.
-Bacteria overgrowth in elder patients in intestine significantly affect absorption.
-Fatty meals generally slow down the absorption but drugs like GRISEOFULVIN easily
absorbed I presence of fatty meal.
-Temperature of meal changes dissolution and absorption.
-Sleeping position slow down absorption
-Stress increases the gastric secretion and slow down absorption.
-Gastroenteritis, Diabetes mellitus, Hypothyroidism retard gastric emptying, whereas
duodenal ulcer, hyperthyroidism promotes gastric emptying.
-Antacid, narcotic analgesics, antidepressants slow down the gastric emptying, but
prokinetics facilitates absorption.
-Disease state like GI infection, cancer, cardiovascular infection, hepatic disease significantly
varies absorption.
 DRUG – DRUG INTERATION GIT:
Antidiarrhoeal preparation containing adsorbant prevent absorption of the drugs.
Antacids, minerals form insoluble complexes of drugs that reduces absorption
Eg: Co-administration of antacid or proton pump inhibitor changes gastric pH on absorption
Anticholinergic drugs retard the mobility of GIT, whereas prokinetics facilitate the emptying
Milk products, grape fruit juices from insoluble complexes totally to prevent insoluble
complex

METHODS TO STUDY GI ABSORPTION

 In vitro ,ex vivo and in vivo techniques are used to evaluate GI absorption of drugs.
 Stimulation of body condition at laboratory levels is called invitro techniques.
 The following in vitro techniques are used to examine GI absorption
1. Diffusion studies : Using egg shell membrane or synthetic membrane between a
donor and receptor compartment and allowing the drug molecule to diffuse through
the membrane are done
Samples are withdrawn from receptor compartment and quantified for drug content
using UV spectrophotometer.
2. Cell culture techniques
 CACO 2 cells {cancerous cells of colon carcinoma} are allowed to grow in a nutrient broth.
 Procedures (like antimicrobial studies) are conducted using drugs in the cells line and
minimum inhibitory concentration are noted and correlated for drug absorption
 Fluorescent markers are also added to examine drug absorption
Exvivo techniques are :
Isolated live tissues of animal or human origin are used in lab setup for studies . This
procedure is called Ex vivo studies

The following studies used for absorption techniques :

i. Intestine portion of animal origin are isolated and used as diffusion. membrane in
the everted position by forming a sac or cutting a rubber band(ring)

Periodical sample withdrawal from receptor compartment and analysing drug content give idea about
absorption

SINGLE PERFUSION, MULTIPLE PERFUSION (Ex vivo studies)

 1. Two extreme end of intestine portion of animals are dissected and attached to peristaltic pump
like dialysis process
2. Perfusion fluids containing drugs are pumped into intestine portion at one end and samples
collected at another end for absorption process

IN VIVO HUMAN STUDEIS

1. Using endoscopic tech. Absorption can be photographed. This method had poor patient complaints
and costly
2. Using markers like mannitol which decrease GI absorption of many drugs can be correlated with
blood level drug
3. Capsule containing isotope technetium(Te99) can be scanned by radiography
4. Remote delivery capsules (Heidelberg capsules) having magnetic response are used to evaluate drug
absorption with help of magnetic field.

PERCUTANEOUS / SKIN/ TRANSDERMAL ABSORPTION

Skin made of 3 layers: epidermal, dermal, subdermal.

Epidermal layer made of toughest keratin, hence called stratum corneaum.

Dermal layer having sweat glands and hair follicles. Blood circulation is prominent in the Layer so called
cutaneous layer.

Below dermal is subdermal layer generally called subcutaneous layer.

Skin is having longer surface area and 1/3rd blood circulation

Skin passes first pass metabolism.

Lipophilic drugs easilygets absorbed through transcellular pathway, hydrophilic drugs follows
paracellular pathway.

Heavy metals absorbs through glands pathway.

Stratum corneaum is very tough area for absorption; so skin contains ketatolytics or surfactants facilitate
absorption by dissolving the layer.

Skin pH ranges between 5-6, external factors like humidity, temperature, microbial attack changes skin
structure and affect the absorption
  Slow release parentrals are injected into subdermal layers .whereas immediate release
preparations are given as dermal injections.
 Iontophoresis (using electric current).sonophoresis (using sound waves) are used to
increase drug permeation through skin. example-Nicotine patches

MUCOSAL ABSORPTION
 Buccal ,sublingual,nasal,rectal,vaginal mucosals are used for drug delivery
 Mucosal route bypass the first pass metabolism and provide similar bioavailability like
IV route
 Injections are to be sterile ,whereas mucosal formulation need not to be sterile
 Mucosal administration favors self administration and increase the patient compliance
 All the hospitals in emergency cases follow inhalation (mucosal)route of delivery

DISTRIBUTION AND FACTORS AFFECTING DISTRIBUTION

 Distribution is reversible transfer of drug between extravascular fluids and tissues.

 Unbound drug molecules depends on rate of perfusion and tissue permeability. It moves from
blood to tissue.
 Tissue permeability mainly depends on physicochemical nature of drugs.
 Drugs having molecular weight less than 50 dalton and water solubility diffuse through aqueous
fluid whereas drugs having higher molecular weight greater than 500 dalton follows transcellular
pathway.
 Molecular weight lesser than 500 dalton drugs are very difficult to diffuse so site specific
formulations are preferred.
 The degree of ionization at blood pH 7.4 plays a vital role in distribution. unionized drug easily
get diffused and lipid molecules get faster distribution. In case of poisoning antidote for
ionization is given .Example acid poisoning treated with sodium bicarbonate.

 Patition coefficient also plays a major role in distribution Ko /w thiopental is a weaker acid
( Ko/w =2) whereas salicylic acid is stronger acid (Ko/w= 0.0005)
 Thiopental shows 80 folds higher distribution than salicylic acid.
 Highly perfused tissues like lungs , kidney, heart, brain mainly depends on perfusion rate in
distribution whereas poorly perfused tisssues like bone and adipose tissues depends protein
binding in distribution.
 High perfused tissues facilitate faster elimination while low perfused tissues accumulate.
 Fat content and total body water: higher in infants so higher distribution is possible in infants.
 Volume of distribution during pregnancy get increased.
 Obesity restrict the distribution and produce accumulation.
 Fatty diet facilitate the absorption of NSAIDs by increasing the distribution.
 MEMBRANE BARRIERS FOR DISTRIBUTION

The following membrane barriers play a major role in distribution

 Capillary endothelial membrane

 Cell membrane
 BBB
 Blood CSF barrier
 Blood placenta barrier
 Blood testis barrier

CAPILLARY ENDOTHELIAL BARRIER:

Molecular weight less than 500D diffuse through this barrier but higher molecular weight
substance are restricted

CELL MEMBRANE BARRIER:

Molecular weight less than 50 d polar drug follows bulk flow where as lipid drugs having molecular
weight less than 500 dalton follows passive diffusion

Ionised drug through active diffusion may be permitted

BLOOD BRAIN BARRIER :

 Brain capillary consists of endothelial cell which are joined to one another by continuous tight
intracellular junction. Presence of spatial cells like astrocytes and pericyte which are supporting
the tissue of endothelial membrane form a solid envelope around the brain capillary.
 As a result passage of polar molecule totally blocked.
 Brain delivery of molecules may be required in CNS disorder. So they molecules are converted to
liphophilic – high Ko / w (partition coefficient)to induce passive diffusion.
 Energy molecule is glucose and amino acid are transported by active diffusion.
 Drugs like penicillin are water soluble never cross blood brain barrier.
 For the treatment of parkinsonism levodopa prodrug is given to deliver dopamine.
 Three different approaches can be used in brain targeting
a) Permeation enhance- dimethyl sulphoxide
b) usage of osmogen (mannitol) – disturb the capillary and allow [permeation.
c) redox system- polar drugs converted as lipid soluble with dihydropyridine.
In presence of CNS enzyme dihydropyridine getting reduced to predinium ions and getting
oxides which cannot diffuse back out of brain.

BLOOD CSF BARRIER:

 Capillary endothelium have open junction as well as tight junction of coroid plexus of glial cells
have tight junction which is similar to brain .
  Highly lipid soluble drug easily diffuse CSF sulphamethaxole and trimethoprim and beta-
blockers have higher rate of CSF distribution.
 Intrathecal injection produce prolong pain because of endothelial rupture.

BLOOD PLACENTAL BARRIER:

 Maternal and fetal blood capillary are separated through placental barrier.
 This barrier consist of endothelium, connective tissue, thromboblast and syndium
 In earlier pregnancy, the membrane used to be in thickness of 25 micrometer but reduce to 2
micrometer during later stage of pregnancy.
 Molecular weight less than 1000 dalton and lipid drug easily diffuse through placental barrier.
Nutrients are transported through carrier mediated transport and immunoglobulin by endocytosis.
 Terotogenic agents produce fetal abnormalities. So during pregnancy the prescription has to be
strictly based on guidelines.

BLOOD TESTIS BARRIER:

Sertoli endothelium act as a barrier in the testies. Alcohol consumption, narcotics and some
categories easily permeates and produces spermicidal effects.

PROTEIN BINDING (DISTRIBUTION)

Complex formulation between drug and extravascular tissue protein are called as protein
binding. After receptor binding drug shows pharmacological response. Binding may be ionic binding or
wanderwaal forces. It binding happens in the blood cells or plasma then it is called plasma protein
binding. If it is happening in the tissue level then it is called tissue localization. Protein binding can be
analysed through filteration or spectroscopic method.

[P] + [D] [PD]

SIGNIFICANCE OF PROTEIN BINDING:

 Plasma protein binding maintain the sink condition for continuous absorption.
 Water insoluble drug heparin, steroids, fat soluble vitamins after binding with become soluble
and distributed throughout body.
 After dissociation of protein complex free drug produce therapeutic response
 Protein binding facilitate the prolonged action.
 Free drug half life can be increased. Eg. Penicillin having short half life after binding with protein
shows prolongation.
 Higher protein bound molecules displaces the digoxin which leads to toxicitizer digoxin. The
displaced toxicity can be minimized by protein binding.
  Displacement can be used as a diagnostic method. Eg. Detection of chloroquine induced
melanoma with radiolabelled I-131.
 Thyroid gland have greater affinity for iodine radioisotope of iodine cane be used for thyroid
disorder.
 Protein binding can be used in drug delivery of hydrophilic moiety.
Eg. Cholesterol binding of anti-cancer drugs used as a target delivery of tumour cells.
 HDL cholesterol mainly concentrate at adrenal glands so nitrogen mustard with HDL cholesterol
used as a targeting device for prostate cancer.

Plasma Protein Binding

Plasma Protein Molecular weight % of Binding Drugs binds

Human Serum 65000 5% All drugs
Albumin (HSA)
α1-acid glycoproteins 44000 0.1% Basic drugs
Lipoproteins >2 lakhs variable Basic lipophilic drugs
α1- globulin 30000 0.001 Starch
α2- globulin 1,40,000 0.05 Fat soluble vitamins
Haemoglobin (Hb) 65000 66 Barbiturates

1) Human Serum Albumin:

It is mostly abundant and binds with all category of drugs. Endogenous fatty acids,
bilirubin, tryptophan, also binds with human serum albumin.

o Site I – Warfarin binding site (NSAID, sulfadrugs) ≥90% drugs binding

o Site II – Diazepam binding site (Benzodiazepines, Ibuprofen)
o Site III – Digoxin binding site
o Site IV – Tamoxifen binding site

2) α1-acid glycoproteins:
Basic drugs like emipramine, propanolol, quinidine binds with α- acid glycoproteins
(orosomucoid)

3) Lipoproteins:
They are classified into 4 categories:

 Chylomicrons
 LDL
 HDL
 Triglycerides
 o Chylomicrons- least dense and larger in size, hydrophilic lipid drugs binds with
lipoproteins and this protein binding is non- competitive.
o Binding of lipoprotein play a major role in distribution of fatty acids in circulation

4) Globulins:
o α1- globulin(transcortin) – Corticosteroids, Cyanocobalamin(Vit.B12), All hormones.
o α2- globulin( ceruloplasmin) – Fatty acids
o β1- globulin(transferrin) – Folic acid
o β2- globulin(carotinoids)
o γ- globulin(Immunoglobulins) – Antigens
o Haemoglobulin – Binding with RBC and carbonic anhydrase enzyme, Hb mainly
participate in oxygen transfer.

TISSUE LOCALIZATION OF DRUGS

● 40% of body weight made up of tissues which is 100% higher than human serum albumin.
● Tissue binding increases apparent volume of distribution of drugs and decreases the required drug
concentration.
● Tissue binding is irreversible which leads to accumulation and toxicity.
● The following order tissue predominates Liver<Kidney<Lungs<Muscles.

1. LIVER:- Halogenated hydrocarbon and paracetamol irreversibly binds and produce

hepatotoxicity.
2. KIDNEY:- Higher protein molecule binds with renal and produce nephrotoxicity..
3. LUNGS:- Basic drugs like Imipramine, Glimepride gets accumulated in the lungs.
4. SKIN:- Chloroquine and phenothiazine accumulate in skin and interact with melanin.
5. EYE:- Chloroquine reacts with Melanin and produce retinopathy.
6. HAIR:- Arsenic and heavy metal produces hair fall and baldness.
7. BONE:- Tetracycline and lead deposit in bone produce discolouration and brittleness in
bones and tooth.
8. FAT(Adipose Tissue):- Pesticide and lipophilic like thiopental accumulate on fatty
tissue and produce severe toxicity.
9. TISSUE NUCLEIc ACID:- Chloroquine binds with DNA and produce teratogenicity.

DISPLACEMENT INTERACTION
● When drug-drug interaction happen of binding site is called displacement interaction.
● Higher affinity drugs displace the lower one sometime it may be beneficial but generally toxic.

★ BENEFICIAL - Warfarin and Phenylbutazone at human serum albumin prolong the anticoagulant effect
of warfarin.
★ TOXIC DISPLACEMENT REACTION - Sodium Salicylate displace bilirubin from albumin site
which crosse blood brain barrier (BBB) and from mental retardation of children.
 EXCRETION

DEFINITION:

It is defined as irreversible transformation of drug and its metabolite form internal to external
environment.

RENAL EXCRETION:

Kidney plays a major role in tye excretion of non-volatile water water soluble water molecules
having 500 dalton molecules weight. Basic unit of renal is nephron. Each kidney has 1 million nephrons.
Nephron is made up of bowmanns capsule, glomerulus proximal tubule, loop of henle, distal tubule and
collecting tubule.

Glomerulus filteration (GFR), active tubular secretion, tubular reabsorption. There are three
process that nephron participates:

Rate of excretion = glomerular filteration + tubular secretion – tubular reabsorption

GLOMERULAR FILTERATION:

It is unidirectional non-selective filteration. Hydrostatic pressure of bowmann capsule is a

driving force. 25% of cardiac output ( 1.2L of blood/min) goes to renal and 120ml/min is filtered through
glomerulus (GFR). If 180l OF substract passing one day through renal only 1.5L is excreted out.

Endogenous amine creatinine (or) exogenous polysaccharide insulin, used as a marker, to

evaluate GFR and kidney function.

TUBULAR SECRETION:

Active tubular secretion, with energy anionic and cationic drug are secreted in separate system.
Tubular secretion depend on Ph and protein binding but GFR depends on blood flow.

Paraamino hippuric acid (PAH acid) and iodo pyrazate used as marker to asses tubular secretion.
Secretion follows competitive elimination for gonococcal infection, penicillin G + probencid to
competitively block and prolong penicillin G action in tubular secretion.\

TUBULAR REABSORPTION:

Tubular reabsorption to maintain the fluid level, tubular reabsorption act as passive or active,
most of the filter molecule got reabsorped. Concentration gradient plays a major role in reabsorption.
Electrolyte, glucose, vitamin are actively reabsorbed some liphophilic substance are absorbed through
passive process.

CONCEPT OF CLEARANCE

clearance is hypothetical volume of liquid containing drugs from which drug is completely removed in a
specific time.

Clearance cl = dc/dt
 C

Cl T = clR + clNR

Rate of Filteration (Gfr) + Rate of Secretion - Rate of Reabsorption

clR=

Plasma Drug Concentration

RENAL CLEARANCE MECHANISM EXAMPLE

0 Filteration followed by complete Glucose
reabsorption
<130 Filtered partially reabsorbed Lipophilic drug
130 Filtration Inulin, creatinine
>130 Filtration + secretion Ionic drug
650 Total secretion PAH

Inulin is exogenous polysaccharide creatine is endogenous amine. Eliminating creatinine is easy, so it is

used as a marker to asses renal function

RF = clcr of patient

Clcr of normal adult

Children (1-18),

Clcr = 0.48H/Scr[ W/70] 0.7

Male ,

Clcr = [(140-Age)/72* serum creatinine]W

Female,

Clcr = [(140-Age)/85* serum creatinine]W

If renal function < 25% , then dialysis either peritonial or hemodialysis to be conducted

If renal function < 10% transplantation is recommended

In case of poisoning using charcoal hemofiltration or perfusion are done.

In renal failure patient dose or dosing interval are adjusted as follows

Dose adjustment in renal failure = normal dose [ fu( RF -1) + 1]

Where,
 Fu = fraction in urine

RF = renal function

Dosing frequency adjustment = normal dosing interval /RF

FACTORS AFFECTING RENAL EXCRETION:

A) MOLECULAR WEIGHT: Drugs having less than 500 dalton molecular weight follows
glomerular filtration whereas higher molecular weight substances eliminated through hepatic
biliary clearance.
B) ACIDIC AND BASIC DRUGS: After getting hydrolysed , eliminated through tubular
secretion.
C) LIPOPHILIC SUBSTANCES: get reabsorbed
D) URINE PH: Acidic drugs in alkaline condition gets completely eliminated whereas basic drugs
get reabsorbed.
E) Blood flow: If blood flow is higher, filtration dominates.
F) PROTEIN BINDING: Protein bound molecule prone to get reabsorbed.
G) Acute and chronic renal function disturbs the normal excretion and toxicity.

NON RENAL EXCRETION:

A) Hepatic biliary clearance(entero hepatic recycling)

B) Pulmonary excretion
C) Salivary excretion
D) Mammillary excretion
E) Skinexcretion
F) Genital excretion

SALIVARY EXCRETION:

Salivary pH varies from 5.8-8.4. Acidic drugs like caffeine, theophylline, phenytoin and carbamazepine.

Phenytoin by passive diffusion gets eliminated through saliva, penicillin gets secreted out through saliva.

Phenytoin produces gingival hyperplasia, sulphonamide and clonidine recycled through saliva

If ph becomes acidic, alkaline drug elimination predominates.

If ph becomes alkaline, acidic drug elimination predominates.

PULMONARY EXCRETION:

Through passive diffusion , volatile substances like halothane, nitrous oxide are eliminated. Carcinogenic
benzene steam distillated in lungs and produces toxicity.

MAMILLARY EXCRETION:
Lactic acid is a extracellular fluid which also has lipoproteins. ph of milk is 7, unionized molecule is
easily secreted through milk, so lactating mothers while using the following medications are advised to
check for the side effects in infants.

a) CHLORAMPHENICOL : bone marrow suppression

b) DIAZEPAM : prolonged secretion
c) HEROIN : neonatal dependence
d) PROPYL THIOURICACID : suppression of thyroid symptoms
e) TETRACYCLINE : staining of infant tooth

SKIN ELIMINATION:

Through sweat gland, benzoic acid , salicylic acid, alcohol passively diffuse out and produce
hypersensitivity.

Heavy metals like lead, mercury, arsenic excreted through sweat glands and produce dermatitis.

BILIARY CLEARANCE
(Entero Hepatic recycling)

DRUGS BILE

SMALL LIVER
INTESTINE

FECAL Hydrolysis
ELIMINATION Mol.Wt. > 500
BLOOD
Daltons

Hepatic cells living the bile duct produces bile acids through active secretion. Secreted bile acid reaches
duodenum and facilitates absorption of fats. If molecular weights are more than 500Da (glucouronide
conjugation) eliminated through faecal way. Hydrolysed molecules are recycled through Entero hepatic
cycling; sulphobromo ofthalene is used as a marker to evaluate Biliary clearance if elimination within half
an hour, an Entero hepatic cycling is normal.

Endogenous substances like Vit B12 , folic acid, steroidal hormones and bile acids are preserved through
Entero hepatic recycling. Sulpha drugs, chloramphenicol, morphine and indomethacine follow Biliary
clearance. Inulin, sucrose, phospholipids follows renal clearance . Na +, K+, Cl – ions have equal ratio
between plasma and bile acid so they equally follows renal and Biliary clearance.
Pesticides like DDT are reabsorbed through entero hepatic recycling. Polystyrene used as antidote to
break recycling and prevent toxicity.

Biliary clearance = Bile acid secretion rate

Plasma drug concentration

PHARMACOKINETICS

It is a science deals with drug absorption distribution metabolism and elimination (ADME). Kinetic
studies can be conducted using following models

In vitro (dissolution apparatus):Ex-vivo (isolated animal tissues): In vivo (animal/human): Insitu/In silico
(tissue culture eg- cancer of colon cells Caco-2 cell line)

In vivo samples may be invasive (blood) or non-invasive (urine)

Pharmacokinetics may be extended to

Clinical pharmacokinetics (application of PK principles for patient clinical management)

Population pharmacokinetics (survey of PK parameters in particular ethnic groups)

Toxicokinetics (evaluation of PK parameters of poison or allergens)

PLASMA LEVEL TIME CURVE

The plot between plasma drug concentration VS time is used to find out the pharmacokinetics and
pharmacodynamics parameters.

PHARMACOKINETIC PARAMETERS

Cmax ,tmax and AUC

Cmax

It is the peak plasma drug concentration.

tmax

The time to reach peak plasma drug concentration.

AUC

It is the area under the curve when the time is in X-axis and plasma drug concentration is in Y-axis.
  The area can be calculated using trepezoidal rule.
 Area = ½ (Y1+Y2) (X2-X1)

PHARMACODYNAMICS PARAMETERS

 Using plasma level time curve the following pharmacodynamic parameters can be obtained.

MEC (MINIMUM EFFECTIVE CONCENTATION)

After reaching MEC, drug shows therapeutic actions.

 The time to reach MEC for a drug is called onset time .

 The ascending phase is absorption phase and the descending pattern shows biotransformation.
 Delayed release formulation (enteric coated tablets) take some time for absorption.
 The time difference between tablet ingestion to the absorption is called lag time.
 Duration of action is the time interval in which the drug maintains the concentration at effective
level.

MSC (MAXIMUM SAFE CONCENTRATION) OR MTC (MINIMUM TOXICOLOGICAL

CONCENTRATION)

 During the Cmax , the therapeutic action is called intensity of action.

 The therapeutic range is the range between MEC to MSC.
 The therapeutic window is the range between MSC to MEC.
 What will be the therapeutic index if the drug shows 50-500µg/ml?
 If the numbers are 2-3, then they are called narrow therapeutic index drugs.

ZERO ORDER KINETICS V/S FIRST ORDER KINETICS

When a drug A converted to metabolite B the change of drug is noted as

A B = -dA / dt

Where as formation of metabolite B,

+dB / dt

In general concentration change

dc/ dt = -kC n

k = rate constant

n = order of reaction

Rate of reaction is defined as change of molecules / unit time.

Order of reaction is defined as the manner in which active ingredient participate in reaction .

When n=0(zero order reaction)

dc/ dt = -k

It is independent on reactant concentration.

When n=1 (first order)

dc/ dt = -kc

It is a reaction depending on concentration of reactant

ZERO ORDER FIRST ORDER

Rate of reaction is independent on reactant Dependent on reactant concentration.
concentration.
dc/dt = -k dc/dt = -kc
concentration at time t concentration at time t
ct = co-k ct = coe-kt
t = c2-c1/k t = Inc2-Inc1/k
In plot In plot

The time taken to get 50% of initial concentration The time taken to get 50% of initial concentration
t1/2 = 0.5c0/k t1/2 =0.693/k
IV infusion, controlled release formulation. All chemical decomposition, biotransformation.

PHARMACOKINETIC MODELS

Kinetic process is a complex one so studying this process using in vivo sample model independent
approaches are used.

A model is defined as hypothetical unit to correlate mathematical terms with quantitative analysis of drug.
So a model gives idea about

1. Behaviour of drugs in tissue

2. Predicting plasma drug concentration
3. Extending single dose to multiple dose
4. Calculation of optimal dosage regimen
5. Toxicity of drug
6. Correlating kinetics with pharmacological response
7. Evaluate bio equivalence
8. Find drug drug interaction
9. Drug influence in phathological change
10. Predicting possible active metabolites
MODEL APPROACH

Three different models

 Distribution model
 Empirical model
 Physiological model

DISTRIBUTION MODEL (Receptor model)

This model assumes even organ contains no. of receptor & each receptor bins with specific drug
after binding, drug receptor complex exhibit therapeutic responses (lock & key therapy).

EMPIRICAL MODELS (COMPARTMENT MODELS)

These are hypothetical in nature. They are arranged in serial or parallel in which they communicate each
other. Each compartment is not a real physiological or anatomical region. Each compartment is
uniformly distributed and follows first order kinetics. Based on arrangement they are classified into :

1. MAMMILARY MODEL (SATELLITE MODEL)

In this model ,the main compartment (central compartment) well connected with peripheral compartment

Highly perfuse tissues like liver, kidney, lungs, heart act as central compartment. While low perfuse
tissues like bones and skin act as peripheral compartment.

Movement from central compartment follows first order kinetics, one compartment model shows only
central compartment model, whereas multi compartment model have central and peripheral compartment

No. of rate constant participate in the event can be given as:

R =2n-1 (IV BOLUS)

R= 2n (other than IV bolus)

N = No. of compartment

One compartment oral :

Ka Ke
central
Absoption rate constant Elimination rate constant

One compartment IV Bolus

central Ke
One compartment IV infusion

Ka central Ke

Rate of infusion

Two compartment IV Bolus

K12 Peripheral
central

Two compartment IV Infusion

K0 central K12 Peripheral

Ka Ke K21
K21
ORAL

DISTRIBUTION CONSTANT

CARTNERY MODEL (RAILCOACH MODEL )

Compartments are arranged in serial manner. This model is considered as basic model to describe
characters of a drug in animal model.

ADVANTAGES:

1. Based on mask balance easy to describe drug movement in animal model.

2. Visual representation of various processes is possible.
3. Easy to find out influencing factors.
4. Each rate process can be easily calculated.
5. Easy to correlate normal physiology and pathophysiology.
6. With limited data extending to maximum population possible.
7. Initial toxic screening possible.
8. Multiple drug analysis possible.
9. Dosage regimen is possible in this model.

DISADVANTAGES

1. Extension of animal physiology& human produce error.

2. To correlate ADME from basic data is very difficult.
3. Multi-exponential relation(Aspirin used as analgesic, NSAID, and anti-platelet) very difficult to
correlate.
4. Different route of administration produces different results.
PHYSIOLOGICAL MODEL

This model describes each kinetic parameter in individual organ and gives perfect data. Based on
equilibrium, organs are classified as:

 Rapidly equilibrating tissues. (RET)

 Slowly equilibrating tissues. (SET)

RET
RE
T
LUNGS

AA VV
HEART
RR
EE
TT
EE LIVER
II
RR
YY KIDNEY NN
SET

BONES AND
SKIN
Based on flow-rate of blood, this model is utilized for input and output rate calculations. Each organ has a
membrane and it acts as a barrier for distribution. So this model is called Perfusion model, Permeation-
rate model or Diffusion-limited model.

ADVANTAGES:

1. Mathematical relation is straightforward to anatomical region.

2. Pathophysiological changes can be correlated with drug response.
3. Mechanism of ADME can be easily explained.
4. Animal and human correlation is possible.

DISADVANTAGES:

1. Time consuming to study

2. Costly
3. Complete sample collections are required.
4. Prediction of individual patient may not be accurate.
 DIFFERENCE BETWEEN PHYSIOLOGICAL & COMPARTMENT

Realistic Hypothetical
PHYSIOLOGICAL COMPARTMENT
Time consuming simple & flexible
Used in advanced stage used in preliminary screening
Mathematics is straight forward easy curve fitting but tough mathematics
Give entire data about individual organ limited data about individual organ
Correlation between human & animal is possible correlation may not be accurate

ADME mechanism is easy to explain explanation is difficult

Pathophysiological changes can be assessed pathophysiological changes not be assessed

NON-LINEAR KINETIC

NON-COMPARTMENT ANALYSIS

It's a model independent approach based on the initial or first movement.the extrapolation done for the
entire spectrum. Statistical moment theory (SMT) is a hypothesis of predicting future response based on
past performance. Scientifically it can be defined that changes in macroscopic event with respect to
residence of a trace molecule and its probable density. This model doesn't require and compartment and
assume drugs and metabolites following the same pattern. Collection of experimental data for a single
dose of drug and extrapolation to the future response with higher doses are generally done

MRT is defined as average amount of time spend by the drug in the body before being eliminated .Using
MRT pharmacokinetics parameters like half life of elimination AUC and bioavailability are calculated.

ADVANTAGES

PK parameters can be calculated using simple algebra

Mathematical relation between drug nas metabolite can be correlated

With limited data entire spectrum of kinetics can be calculated

DISADVANTAGES

Humans are having inter -intra subject variation .these model cannot predict those variations

This method not applicable for non linear kinetics

 Non-linear kinetics

Linear Kinetic are following first order kinetic and superimposition principle increased in
concentration directly proportional to increased the effect. Nnon- linear linetics do not follow first order
kinetic instead it follows mixed order of first order and zero order.

Emergency medicine (narrow therapeutic drugs) generally follows non-linear pathway. Micheals
and Menton derived the equation for enzyme saturation kinetics.

(rate of changes) -dc/dt = Vmax C/ Km+ C

Vmax = maximum velocity

C= concentration of drug

Km= Michealis Menton rate constant

1) If Km > > C
-dc/dt = Vmax X C / Km (C negligible, so first order or concentration first order )

2) If Km = C= 1

-dc/dt = Vmax . C/C = Vmax (zero order) , high concentration zero order

CAUSES FOR NON-LINEARITY:

ADME are prone to follow enzyme- saturation kinetics.

1) ABSORPTION LEVEL:
Eg: insoluble drugs like griseofulvin because of lower solubility and dissolution easily get
saturation at GIT.
Eg: Water soluble vitamins follows enzyme carrier pathways and leads to non-linear absorption.
Eg: first pass metabolite drug [ pre-systemic elimination]
Eg: propanolol hydrochloride gives poor bioavailability.
2) DISTRIBUTION LEVEL:
After reaching blood circulation drug binds with plasma tissue. Pheylbutazone (NSAIDs) binds
with plasma level.
Saturation of plasma protein leads to higher volume of distribution.
Thiopentol and fentanyl binds at tissue level and reduce the volume of distribution.
3) METABOLISM LEVEL:
Carbamazepine at higher dose induce enzyme metabolism which leads to autolysis.
Phenytoin, theophylline and alcohol having capacity limited metaboilism. Higher doses leads to
higher concentration of study state (drug tolerance)
4) EXCRETION LEVEL:
Eg:
1) Penicillin G and probrncid both follows active tubular secretion. Probencid decreases the
clearance of penicillin G and prolongs antibiotic effect (beneficial drug interaction)
 2) Water soluble vitamins and glucose are reabsorbed to tubular saturation that leads to
elimination.
3) Tetracycline, indomethacin eliminated through bile duct and saturation leads to toxicity.

Estimation of Michalis Menton constants (Km,Vmax)

BIOAVAILABILITY:

It is the rate and extent of drug reaches into systemic circulation ( absorption).

BIOAVAILABILITY (F) = BIOAVAILABLE DOSE/ ADMINISTERED DOSE.

IV BOLUS:

It bypass the first pass metabolism and provide absolute bioavailability.

METHOD FOR ESTIMATING BIOAVAILABILITY:

NDA ( new drug application)

ANDA ( abbreviated new drug application)

NDA and ANDA requires bioequilance studies in healthy human volunteers (clinical trial). The
bioavailability estimation are done as follows:

1. PK method

Plasma level time curve

Urine estimation curve

2. PD method
3. Therapeutic equilance
4. Invitro dissolution

URINE ESTIMATION CURVE:

Cumax- cumulated amount of drug in urine

Tumax- time to get maximum drug in urine

PD METHOD:

Pharmacological response in chart like EEG,ECG are compared between sample and reference.

THERAPEUTIC EQUIVALENCE:

Therapeutic response from the patient are taken as feedback for sample and reference.
 INVITRO DISSOLUTION:

Pharmacokinetic dissolution apparatus are simulating GI environment. Oral solid formulation are
conducted for dissolution.( dissolve completely)

Periodical samples are withdrawn and analysed by uv spectrophotometer for drug content(invite
release)

BIOPHARMACEUTICAL CLASSIFICATION SYSTEM (BCS)

Based on solubility and permeability drugs are classified into 4 types

TYPE I : dissolution is proportional to bioavailability.

TYPE II : 0% dissolution and 100% bioavailability. Low soluble drugs require solubility enhancement.

TYPE III: 100% dissolution and 0% bioavailability. Low permeable drugs require permeability
enchancer.

TYPE IV: 0% dissolution and bioavailability . only IV form is available.

IV I

LS , LP HS , HP

eg : peptide hormones eg: diclofenac sodium

III II

HS , LP LS , HP

eg: ranitidine Hcl eg: nimusulide

LS - low soluble

LP –low permeable

HS – high soluble

HP- high permeable

INVITRO IN VIVO CORRELATION (IVIVC)

To minimize clinical study invitro dissolution studies are used as an alternate (surrogate) for batch to
differentiation . This IVIVC is correlation to minimize.
 INVITRO INVIVO
t50% (time to release 50% drugs) AUC , Cmax
t90% Ka , F (fraction bioavailability)
MDT MAT
Cumulative % drug release Xu
t50% LD 50 (or) ED 50

LEVEL OF CORRELATION

 Level A (USFDA recommended)

Invitro drug release at time 0-8 Vs in vivo parameters 0-8 hrs

 Level B (average)
MDT Vs MAT
 Level C ( single point)
t 90% Vs Ka

BIOAVAILABILITY ENHANCEMENT THROUGH SOLUBILITY AND DISSOLUTION

Type II nd type III drugs are having bioavailability issues.
Type II drugs require solubility enhancement.

TYPE II ( solubility and dissolution enhancement)

i. MICRONIZATION : eg :Griseofulvin solubility enhanced by size reduction.
ii. NANO SUSPENSION: eg : amphotericin B converted as nanosize particle using homogenizer.
iii. SUPER CRITICAL FLIUD RECRYSTALLIZATION (GREEN CHEMISTRY) : using Co2
drug particle recrystallized to smaller size.
iv. SPRAY FREEZING : using helium gas
v. EVAPORATIVE PRECIPITATION : eg spironolactone (converted into small size).
vi. SURFACTANT : steroids
vii. SOLID DISPERSION.
viii. BETA CYCLODEXTRIN COMPLEX : converted into solubilizing form. eg: barbiturates.
TYPE III (permeability enhancement)
i. Liquid suspension
ii. Emulsion
iii. Solid-liquid nanoparticle
iv. Liposomes
v. Ion – exchange resin complex
vi. Addition of lipid permeation enhancer.

BIOEQUIVALENCE
In basic, moiety salt are different ( eg: diclofenac sodium, diclofenac potassium) . they are called
chemical equivalent. Same drug in different brandsor formulation (eg: diclofenac tablets and capsules). If
the drugs are from same therapeutic category then they are called as therapeutic equivalence ( diclofenac
and aceclofenac).

In rate and extent of absorption are similar for 2 products of same drug. They are called
bioequivalent

US FDA recommended bioequivalent studies in following conditions.

 Narrow therapeutic index drugs

 Changes in excipients level
 Changes in stereochemical nature of drug
 Change in storage condition
 Change in manufacturing process

BIOWAVIER

It is the process of exempting biequivalence study. Condition for bioequivalence are

 If 2 formulation of a drug show similar dissolution profile

 Changes only in diluent

BIOEQUIVALENCE STUDY PROTOCOL DESIGN

TITLE :

 Principle investigator
 Project number and date

STUDY OBJECTIVE

STUDY DESIGN

 Designs
 Drug product and reference
 Dosage regimen
 Sampling schedule
 Housing
 Meals

STUDY POPULATION

CLINICAL PROCEDURE
  Dosage administration
 Sampling
 Activity of subject

ETHICS

 Principle
 Institution review board
 Inform consent
 Indication for withdrawal
 ADR in emergency

FACILITIES

DATA ANALYSIS

 Techniques
 Statistics

ACCOUNTABILITY

 Narcotics

ANNEXURE

 Schedule Y

STUDY DESIGNS

EXPERIMENTAL

A) Meta analysis
B) Randomized block design
C) Randomized controlled design
D) Repeated measure cross over design
E) Latin square design

OBSERVATIONAL

A) Cohort study
B) Cross sectional survey
C) Case control study
D) Case report

EXPERIMENT

A) META ANALYSIS
 Systemic overview of therapeutic issue

B) RANDOMIZED CONTROLLED DESIGN

In experimental , all treatments are randomly alloted to all subjects
Eg: 1 3 5 7 9------A
2 4 6 8 10----B
Advantages :
Easy to conduct an analysis
Disadvantage
All subjects are homogenous, error may be there.
C) RANDOMIZED BLOCK DESIGN
Homogeneous groups are allloted as blocks and treatment assigned randomly within the block .

Advantages :
More number of treatment can be conducted at same time and if dropping are there new
subject can be added in same block

Disadvantage :

Homogeneous group can be baised.

D) REPEATED MEASURE CROSS OVER DESIGN

Two or more treatments are given one after the another in the same group of patients
known as repeated measure cross over design.

Eg :

1 treatment 2nd treatment

A B
Advantages :
Variation are minimum and cost effective method
Disadvantage :
The order of the treatment may have carry on effect
Proper washout period should be given

URINARY DATA ANALYSIS

In absence of plasma data, urine samples are best substituent to calculate pharmacokinetic parameter like
volume of distribution Vd , clearance Cl , elimination rate constant Ke , half life t1/2 .

 Non invasive technique ( no need of sterile procedure)

 No need of technical assistant to collect sample.
 Low cost analytical techniques can be used.

LIMITATION

 Total sample volume to be collected without wastage.

 Collection time to be noted by patient.

CRITERIA:

 Each sample should have 10%of parent drug.

 Analytical technique should differentiate drug and its metabolite.
 Before sampling, bladder has to be emptied.
 200ml fluid with medicine and after 2 hour 200ml fluid should be given to patient to maintain
micturation.
 Timing and volume of urine sample to be noted.
 Change in colour or pH makes error, so it has to be noted.
 Upto 7th t1/2 of the drug, sample should be collected.

RATE OF EXCRETION ARE ( AMOUNT REMAINING TO BE

EXCRETED)
(+) no need of sampling upto 7th t1/2 (log data) (+) more accurate result
(-) points in graph overlaps, calculation may (-) sampling for SR tablet upto 7 th t1/2 , time
not be accurate. consuming.

ONE COMPARTMENTAL IV BOLUS OPEN MODEL

(Instant distribution model)

Hospitalised patient during emergency generally prescribed as iv bolus dose because of instant
distribution and absolute bioavailability of drugs .

The distribution leads equal distribution throughout the body unidirectional; elimination called as open
model.

Iv bolus

One compartment Ke

First order kinetics

Pharmacokinetics parameters which are closely related to patient physiological mechanisms are called as
primary parameters.

Example :volume of distribution, clearance time, extraction ratio

Secondary parameters are derived parameters mainly related to drug.

Example :Ke, t1/2

dx/dt =-KeXo

Ct=Co *e-Ket
logCt=logCo –Ket/2.303

t1/2 =0.693/Ke

Primary parameters

APPARENT VOLUME OF DISTRIBUTION

Apparent volume of distribution is defined as hypothetical volume of body fluids in which drug is distributed .

V = amount of drug(x)
d

concentration of drug(c)

C=x/v

100ml

100ml 100ml

500ml water 500ml water

+1gm charcoal +1gm charcoal
500ml water +1gm cotton
sponge

Consider three beakers having volume of 500ml each

In second beaker B if 1gm charcoal is added to maintain the same menincus another 100ml water is
added.

In third beaker if 1gm charcoal and 1gm cotton sponge is added then 200ml additional water is added to
maintain the level .charcoal acting as adsorbent whereas cotton sponge acting as absorbent .
Human body having 5-6litres of blood looks like beaker A whereas beaker B indicates the presence of
extracellular fluid (12litre ) . Beaker C imitates total body water i.e vascular ,extracellular and
intracellular fluid (42litre). To find out plasma volume Evans blue ,indocyanine green and iodine 131
albumin are used as markers.To check the extracellular fluid volume inulin ,mannitol, and sodium
chloride are used as markers.To determine the intracellular fluids D2O and antipyrine used as markers .

Drugs distributed in blood level Example: warfarin having volume of distribution 10litre whereas
antimalarial drug chloroquine having volume of distribution 15000litres

Drugs having low volume of distribution easily gets eliminated but higher volume of distribution of drugs
leads to accumulation and toxicity

While deciding dosage regimen physicians are adviced to prefer drugs having low volume of
distribution.

Vd=FXo/KeAUC

EXTRACTION RATIO

It is an index showing how efficiently eliminating organs clear blood flow which contains drugs .

ER=Cin-Cout

Cin

Cl=Q.ER

Q=BLOOD FLOW

F=1-ER

Drugs having high extraction ratio (above 0.7) mainly depends on perfusion rate .

Example: propranolol hcl

Drugs having low ER (LESS THAN 0.3) depends on protein binding

Example;theophylline hcl

Liver is having higher ER indocyanine green is used as marker to asses it.

Enzymes play a major role in extraction .Each organ has inherent ability to remove xenobiotics which is called intrinsic capacity clearance

One compartment IV infusion:

IV bolus dose may produce precipitation to overcome these IV infusion generally given. Chronic
pharmacotherapy and chemotherapy with parental nutrition are recommended as IV infusion.
 R = 2n no of compartment
No of rate
Body
Ro Ke constants R = 2(1)=2

e-ket

e-ket

e-ket

e-ket

Plantean

input = output
 Infusion stopped
Cp
Ro

Time
e-ket

IV Bolus
 MSC

IV infusion

Cp
MEC
Fast dissolving tablet

Time

To reach instant effective plasma conc, drugs are given as dissohring tablets or IV bolus injection. This
dose is called priming dose or loading dose.

TWO COMPARTMENT IV INFUSION

Ro k12 (organ)
Central compartment
Rate of (blood) k21 Peripheral compartment

Infusion (internal distribution constants)

Ro=rate of infusion,

Ke=elimination rate constant,

Rate constant: R=2n = 2(2)=4

Organ specific drugs generally follows two compartment system.

Eg: beta blockers IV infusion is preferred in chronic maintenance of therapy without producing
precipitation toxicity.

Concentration of drug (ct)=Ro/Vc Ke [1+[(Ke-β)/(β-α)]e-αt +(Ke-α/α-β)e-βt]

elimination distribution

vc=volume of drug in central compartment,

β= elimination constant,

α=distribution constant,

ke=total elimination constant.

steady state concentration(css)=Ro/Vdβ=Ro/ClT=INPUT/OUTPUT

LOADING DOSE: XO,L=Css.Vc=Ro/Vc.β ×Vc=Ro/β

if β=Ke then Vd=Vc.

TWO COMPARTMENT IV BOLUS MODEL

IV bolus route is considered as absolute bioavailable so it is preferred during emergency highly perfused
tissues act as central compartment where as low perfused tissues act as peripheral compartment.

Though brain is highly perfused due to blood brain barrier it is considered as peripheral compartment.

R = 2n – 1

If n=2

R = 2 x 2-1

R=3

K12

CENTRAL COMPARTMENT PERIPHERAL COMPARTMENT

K21

Ke

ELIMINATION
Organ specific drugs generally follows two compartment systems.

The concentration of rug in central compartment

dCp/dt= K21 Cp – K12 Cc – Ke Cc

drug concentration in peripheral compartment

dCp/dt = K12 Cc – K21Cp

Cc = Ae-αt + Be-βt

Where,

Ae-αt – distribution

Be-βt – elimination

α , β- hybrid constants
K12 , K21- micro transfer constant

C = Be-βt(during elimination , Ae-αt = 0 )

Cr = C-C = Ae-αt (distribution)

Co = A + B

Ke = αβCo/Aβ + Bα)

K12 = AB(β-α)2/Cc (Aβ + Bα)

K21 = Aβ + Bα/Co

AUC = A/α + B/β

Vd = x/Ke. AUC

Vp = Vc. K12 / K21

ClT = β .Vd

ONE COMPARTMENT ORAL ADMINISTRATION (EXTRAVASCULAR MODEL)

 Oral route is the most preferred route for patient due to self administration and painless delivery.

 More than 75% pharmaceutical products are oral products.

 Extravascular induces oral and mucosal, but mucosal routes are having faster adsorption.
 Oral route is having higher residence time than mucosal route and having advantage of lesser
irritation.

 Oral adsorption may follow zero order kinetics if the products are controlled drug delivery
systems but conventional products follows first order adsorption.

 Plasma level time curve of oral route shows biphasic pattern, ascending curve upto Cmax
followed by descending one during elimination.

 Oral route used to have log time.

 Log time is the interval between time of application to starting of absorption.

 One compartment model for extravascular administration has two rate constants.

Determination of Elimination rate constant:

 By semilog lpot between plasma drug concentration Vs time shows linear descending phase with
a slope value of –Ke/2.303

 The rate of process dx/dt,

dx/dt = KaXa – KeX where;

 Ka – First order absorption rate constant

Xa _ Amount of drug to be absorbed
Ke _ Elimination rate constant
X _ Elimination amount at time ‘t’.

 After integration and substitution, plasmadrug concentration at time ‘t’.

C = KaFX/vd (ka – ke) [e–ket- eKat] where:

F – fraction oral bioavailable

Xo _ initial dose
Vd _ volume of distribution

 At Cmax , input = output

Ke . e-Ket = Ka . e-Kat

 Converting into logarithmic,

 Log Ke -Ket/2.303 = log Ka – Kat/2.303

 At Cmax , time t is max.

t max = 2.303 log(Ka/Ke)/Ka – Ke

 Cmax = 0.37FXo/Vd

 Log time is calculated by,

Log time = 2.303 log(A/B)/Ka – Ke where:

A – Intercept of Ka plot
B – Intercept of Ke plot

 AUC = B/Ke – A/Ka

Cl = FXo/AUC

 Slower absorption and slower elimination lead to prolonged tmax.

 Elimination significantly change AUC than absorption.

PLATEAU PRINCIPLE

 Low dose and prolonged duration never reach effective level. Higher dose and higher frequency
leads to toxic level and may be fatal.
 Normal dose with normal frequency during multiple dosing after fifth half-life of the drug will
become plateau.
 Accumulated concentration is called as UPPER ASYMPHTODE. Elimination concentration is
called as LOWER ASYMPHTODE.
 After fifth half-life of the drug most of the drugs follows steady state principle so prepare dose
adjustment is required to be done after fifth dose.

DOSAGE REGIMEN : (MULTIPLE DOSING)

Acute and Chronic illness required multiple dosing instead of single one.Therapeutic objective is to cure
the illness with shortest time with minimum side effect by the use of least amount of time.

Dosage regimen is a manner drug dosage is given optimal multiple dosage regimen is the one in which
drug is administered in suitable dose by suitable route with sufficient frequency that ensures maintanence
of plasma drug concentration within therapeutic without fluctuation and accumulation for entire duration
of therapy.
Chemotherapeutic and narrow therapeutic index drugs are required proper dosage regimen. If not leads to
resistance and toxicity.

APPROACHES FOR DOSAGE REGIMEN:

1. Empirical
2. Population Average
3. Individualized

1)EMPIRICAL: Physician decide dosage regimen based on previous experience. It may be biased, not
accurable.

2) POPULATION AVERAGE: Based on population pharmacokinetic parameters, average dose is

calculated. T?his may be fixed model, irrespective of ethnicity, normal dose is fixed dose.
ADAPTIVE DOSE: Considering the parameters like age, gender, surface area, BMI, dose is adjusted to
the patient.

3)INDIVIDUALIZED: Cytochrome P450 of enzyme varies from patient to patient.This variation leads to
different levels of metabolism. ( Pharmacogenomics).

Tailoring medications according to individual requirement is the ideal scientific approach. It may be
costly but gives 100% therapeutic outcomes. (Therapeutic drug monitoring)

TWO COMPARTMENT ORAL MODEL

(LOO- RIEGELMAN METHOD)

R = 2n = 2(2) = 4

C= Ne-Kat + Le-Kdt + Me-Ket

LIMITATION:-
1. Simultaneously IV bolus model is required to calculate value (Pharmacokinetic).
2. It cannot be extended to Wagner nelson method.
Ka = Absorption rate constant.
Kd= Distribution rate constant.
Ke= Elimination rate constant.