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CONTENTS
Introduction
Linear & Nonlinearity Pharmacokinetics
Detection of non-linearity in pharmacokinetics
Causes of nonlinearity
Michaelis – Menten equation
Estimation of Km and Vmax #
LINEAR PHARMACOKINETICS
At therapeutic doses, the change in the amount of drug in
the body or the change in its plasma concentration due to
absorption, distribution, binding, metabolism or excretion,
is proportional to its dose.
Whether administered as a single dose or as multiple
doses.
In such situation the rate processes are said to follow first
order or linear kinetics and all semilog plots of C Vs t for
different doses when collected for dose administered, are
superimposable.
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»The important pharmacokinetic parameters viz.
F, Ka, KE, Vd, ClR, ClH
which describes the time course of a drug in the body remain
unaffected by the dose.
» Pharmacokinetics is dose independent.
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NONLINEAR PHARMACOKINETICS
» The rate process of drug’s ADME are depend upon carrier
or enzymes that are substrate specific, have definite capacities
and are susceptible to saturation at a high drug concentration.
» In such cases, an essentially first-order kinetics transform
into a mixture of first-order and zero-order rate processes and
the pharmacokinetic parameters are changed with the size of
the administered dose.
»Pharmacokinetics of such drugs are said to
be dose dependent.
Terms synonymous with it are mixed-order, nonlinear
and capacity-limited kinetics.
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DETECTION OF NON-LINEARITY IN
PHARMACOKINETICS
• There are several tests to detect non –linearity in
pharmacokinetics but the simplest ones are:
1) First test:- Determination of steady state plasma
concentration at different doses.
2) Second test:- Determination of some important
pharmacokinetic parameters such as fraction
bioavailability, elimination half life or total systemic
clearance at different doses of drug.
Any change in these parameters is indicative to non-
linearity which are usually constant.
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CAUSES OF NON-LINEARITY
Drug absorption
• Three causes:- I) Solubility / dissolution of drug is rate-
limited; Griseofulvin - at high concentration in
intestine.
II) Carrier - mediated transport system; Ascorbic
acid - saturation of transport system.
III) Presystemic gut wall / hepatic
metabolism attains saturation; Propranolol.
• These parameters affected F, Ka, Cmax and
AUC.
• A decrease in these parameters is observed in former two
causes and an increase in latter cause.
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Drug distribution
At high doses non-linearity due to
• Two causes:- I) Binding sites on plasma proteins get
saturated; Phenylbutazone.
II) Tissue binding sites get saturated.
• In both cases there is increase in plasma drug
concentration.
• Increase in Vd only in (I)
• Clearance with high ER get increased due to saturation of
binding sites. #
Drug metabolism
• Non-linearity occurs due to capacity limited metabolism,
small changes in dose administration - large variations in
plasma concentration at steady state - large intersubject
variability.
• Two imp causes:- I) Capacity - limited metabolism - enzyme
&/
cofactor saturation; Phenytoin, Alcohol.
II) Enzyme induction - decrease in plasma
concentration; Carbamazepine.
• Autoinduction in dose dependent concentration.
• Saturation of enzymes - decrease in ClH - increase in Css.
• In case of enzyme induction reverse condition.
• Other reasons includes saturation of binding sites, inhibitory #
effects of the metabolites on the action of enzymes.
Drug excretion
• Two active processes which are saturable,
I)Active tubular secretion - Penicillin G
II)Active tubular reabsorption - Water soluble
vitamins & Glucose.
• Saturation of carrier systems - decrease in renal clearance in
case of I & increase in II. Half life also increases.
• Other reasons like forced diuresis, change in urine pH,
nephrotoxicity & saturation of binding sites.
• In case of biliary excretion non - linearity due to saturation -
Tetracycline & Indomethacin.
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Examples of drugs showing nonlinear pharmacokinetics
Causes Drugs
GI absorption:-
Saturable transport in gut wall Riboflavin, Gabapentin
Saturable GI decomposition Penicillin G, Omeprazole
Intestinal metabolism Propranolol, Salicylamide
Distribution:-
Saturable plasma protein binding Phenylbutazone, Lidocaine
Tissue binding Imipramine
Metabolism:-
Saturable metabolism Enzyme Phenytion, Salicylic acid
induction Metabolite inhibition Carbamazepine
Renal elimination:- Active Diazepam
secretion Tubular reabsorption
Change in urine pH Para- aminohippuric acid
Ascorbic acid, Riboflavin
Salicylic acid, Dextroamphetamine
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• Example of Phenytoin
https://www.youtube.com/watch?v=RO4nALnWQtk
youtube.com/watch?v=UvnBmvVyD0M
https://www.youtube.com/watch?v=90noN
X9XMy0
For numerical
https://www.youtube.com/watch?v=DfjyFfr-
mus
MICHAELIS MENTEN ENZYME
KINETICS
It is also called as Capacity-limited metabolism or
Mixed order kinetics.
E+D ED E +M
Enzymes usually react with the substrate to form enzyme
substrate complexes; then the product is formed. The
enzyme can go back to react with another substrate to
form another molecule of the product.
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MICHAELIS MENTEN EQUATION
• The kinetics of capacity limited or saturable processes is best described
by Michaelis-Menten equation.
dC Vmax . C
= ……………….. I
dt KM+ C
Where ,
-dC/dt = rate of decline of drug conc. with time
Vmax = theoretical maximum rate of the
process
KM = Michaelis constant
• Three situation can now be considered depending
upon the value of Km and C.
1) when KM = C:
under this situation , eq I reduces to,
•-dC/dt = V /2 ...................II
max
•
The rate of process is equal to half of its maximum rate. #
•This process is represented in the plot of dc/dt vs. C. shown in fig. 1
2) If a drug at low conc. undergoes a saturable biotransformation
then KM>>C:
• here , KM +C =KM and eq. I reduces to,
-dC/dt =Vmax C /KM………………III
• above eq. is identical to the one that describe first order elimination of
drug, where Vmax/KM= KE.
3) When KM<<C:
•Under this condition ,K M +C= C and eq. I will become,
-dC/dt =Vmax …………….IV
above eq. is identical to the one that describe a zero order process i.e.
the rate process occurs at constant rate Vmax and is independent of
drug conc.
E.g. metabolism of ethanol #
Zero order rate at high doses
Mixed order rate at
intermediated doses
Dc First order rate at low doses
dt
C
Figure 1
A plot of
MME
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ESTIMATION OF Vmax & Km
In enzymatic kinetic work, the classic Michaelis-Menten
equation:
V = Vmax
KM +. C ………..(1)
C where, V= reaction rate,
C= substrate conc. both are used to determine Vmax
& Km.
The velocity of the reaction(V) at various
concentration levels of drug(C) are determined either
by in-vitro experiments or in-vivo experiments at
constant enzyme levels.
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Method 1
By reciprocating equation (1), we get :
1 = Km . 1 + 1 ……..(2)
V Vmax . C Vmax
When 1/V is plotted against 1/C, a straight line is
obtained with a slope of Km/Vmax and an Intercept of
1/Vmax.
E.g. : A plot of 1/ V vs 1/ C (shown in the fig. 2)
gave an intercept of 0.33µmol and a slope of 1.65, Now,
calculate Vmax and KM.
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Km/Vmax
1/ Vmax
-1/ Km
Figure 2
Plot of 1/V vs 1/C for determining Km & Vmax
Now, Intercept= 1/ Vmax = 0.33 µ mol.
Vmax = 3 µ mol/ml min
Slope = Km/ Vmax So, 1.65 = Km/ Vmax Km
= 1.65 Х 3 = 4.95µmol/ ml
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X-axis intercept= -1/ Km
Method 2
Multiplying eq. 2 by C, we
get :
C = Km+ C ………..... (3)
V Vmax
Vmax
A plot of C /V vs C gives a straight line with 1 / Vmax as the slope
and Km / Vmax as the intercept (shown in the fig. 3).
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ax
ax
Figure 3.
Plot of C/V vs C for determining Km & Vmax
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Method 3
The equation can also be written as :
V = - Km V + Vmax
…………(4)
C
A plot of V vs V / C gives a straight line with a slope of –Km & an
Intercept of Vmax. (shown in the fig. 4)
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Figure 4
Plot of V vs V / C for determining Km & Vmax
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CALCULATION OF KM & VMAX
STEADY- STATE CONCENTRATION
• If drug is administered for constant rate IV infusion/ in a
multiple dosage regimen, the steady-state conc. is given in terms
of dosing rate (DR):
DR = Css ……………….. (1)
ClT
• If the steady-state is reached, then the dosing rate = the rate of
decline in plasma drug conc. & if the decline occurs due to a
single capacity-limited process then eq. I become as:
Vmax Css
DR = ……………….. (2)
KM+ C ss
• From a plot of Css vs. DR, a typical curve having a shape of
hocky-stick is obtained which is shown in fig. 5. #
Curve for a drug following nonlinear kinetics
By plotting the steady-state concentration against dosing
rates
Css
Km
Vmax / 2 Vmax
DR (in mg/hr or mg/day )
Figure 5
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METHODS USED TO
DETERMINE THE
KM & VMAX AT
STEADY-STATE
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• There are three methods which are used to define the K M & Vmax
at steady-state with appreciable accuracy:
1) Lineweaver-Burk Plot:- the reciprocal of eq. (2) we get
1 KM 1
= ……………….. (3)
DR Vmax Css +
Vmax
• If 1/DR is plotted against 1/Css a straight line is obtained
having slope KM/Vmax & y-intercept 1/Vmax.
2) Direct linear plot:-
• Plotting a pair of Css, i.e.Css1,&Css2 against corresponding
dosing rates DR1 & DR2 we get following fig. 6 which
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gives values KM &Vmax
Direct linear plot for estimation of KM & Vmax
at steady-state conc. Of a drug, when it is
administered at different dosing rates
DR
Vmax
DR1
DR2
KM
Css 1 Css 2
0 KM
Css
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Figure 6
3) Graphical
method:-
DR
DR/Css
Figure 7
Plot of DR vs DR/Css for determining Km &
Vmax #
3) Graphical
• method:-
In this method by rearranging eq. (2) we get
KM DR ……………….. (4)
DR = Vmax
Css
-
• In graph DR is plotted against DR/Css, a straight line is
obtained with slope –KM & y - intercept Vmax.
•K M & Vmax can be estimated by simultaneous eq. as
DR1 = Vmax - KMDR1 …………….…...(5)
Css1
……………….. (6)
DR2 = Vmax -
KM DR2
Css2
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• On solving above eq. 5 & 6 we get,
DR2- DR1
KM = DR1 DR2 ……………….. (7)
-
Css 1 Css 2
• By substituting values of DR1, DR2, Css1 & Css2 we get value of KM &
from KM we can found value of Vmax at steady-state
concentration.
•
From experimental observations, it shows that KM is much less
variable than Vmax.
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REFERENCE
1. Biopharmaceutics and Pharmacokinetics a treatise by
Brahmankar DM, Jaiswal SB.
2. Biopharmaceutics & pharmacokinetics by Dr. Shobha Rani
R. Hiremath.
