Guided By: Submitted By:

Prof. Preeti K Suresh Madam Harsha Farenjiya Khatik

 Introduction

➢ The process of biotransformation or metabolism is the enzymatic conversion of a drug to a metabolite.

➢ It explains the enzymatic reaction rate of change(velocity) of plasma drug concentration.

➢ It is used to describe non-linear pharmacokinetics.

➢ Non-Linear Pharmacokinetics:
• Follows Mixed-order kinetics.
At lower dose, drug shows first order kinetics but at higher dose, it shows zero order due to saturation, so it is
known as mixed-order kinetics. (eg. Phenytoin or ethanol).
• It doesn’t follow first-order kinetics as the dose increases.
• It may result from the saturation of an enzyme or carrier mediated system.
• The AUC is not proportional to the dose.
• The amount of the drug excreted in the urine is not proportional to the dose.
 Introduction

The enzyme interacts with the substrate to form an enzyme-substrate complex, which leads to
synthesis of the product and the release of the enzyme :

k1 k3
E + [S] ES E + P
k2

Where E , S and P are enzyme, substrate and product, respectively.

 Assumptions

• Total enzyme concentration remains constant during the reaction.

• Amount of enzyme is very small compared to amount of substrate.

• The product concentration is low so that the product inhibition is negligible.

• The reversible reaction P S is not considered because the equation describes initial rates
when [ P] is near zero.

• The ES complex is a STEADY STATE INTERMEDIATE i.e. the concentration of ES remains

relatively constant because it is produced and broken down at the same rate.

Hence, The rate of an enzymatic reaction is dependent on the concentrations of both the enzyme and
the drug and that an energetically favored drug–enzyme intermediate is initially formed, followed by
the formation of the product and regeneration of the enzyme.
Mathematical Expression of M-M Equation :

As per the M-M equation, the velocity (v) of metabolism is given by the equation :

Vmax [S]
v =
Km + [S]

Where,
v = Forward rate of decomposition of the enzyme–drug [ES] intermediate
Vmax = Maximum Velocity of the reaction.
S = Substrate Concentration
Km = Michaelis constant
 Relationship between Rate of Reaction & [S]

For an enzyme-catalysed reaction, there is usually a hyperbolic relationship between the rate of reaction and
the concentration of substrate, as shown below:

B
Relationship between Rate of Reaction & [S]

C (A) At low concentration of substrate, there is a steep increase in

the rate of reaction with increasing substrate concentration. The
catalytic site of the enzyme is empty, waiting for substrate to
B bind, for much of the time, and the rate at which product can be
formed is limited by the concentration of substrate which is
available.

There are abundant enzymes to catalyze the reaction, and the

rate of metabolism is a first-order process.

→ Km = [ S ]

→ v = Vmax/ 2
Relationship between Rate of Reaction & [S]

C (B) Concentration of the substrate increases, rate of the reaction

increases but not in a directly proportional way like previous
phase.
B
It doesn’t follow first-order kinetics as the dose increases.

Hence, This region is comprise of Mixed-order kinetics.

→ Km >> [ S ]

→ v = Vmax . [ S ]
Km
Relationship between Rate of Reaction & [S]

C (B) As the concentration of substrate increases, the enzyme

becomes saturated with substrate. As soon as the catalytic site is
empty, more substrate is available to bind and undergo reaction.
B The rate of formation of product now depends on the activity of
the enzyme itself, and adding more substrate will not affect the
rate of the reaction to any significant effect.

Saturation of the enzyme usually occurs when the plasma drug

concentration is relatively high, all the enzyme molecules
become complexed with drug, and the reaction rate is at a
maximum rate; the rate process then becomes a zero-order
process.

→ Km << [ S ]

→ v = Vmax
Relationship between Rate of Reaction & [S]

The rate of reaction when the enzyme is saturated with

substrate is the maximum rate of reaction, Vmax.

The relationship between rate of reaction and concentration of

substrate depends on the affinity of the enzyme for its
substrate. This is usually expressed as the Km (Michaelis
constant) of the enzyme, an inverse measure of affinity.

For practical purposes, Km is the concentration of substrate

which permits the enzyme to achieve half Vmax.

An enzyme with a high Km has a low affinity for its substrate,

and requires a greater concentration of substrate to achieve
Vmax.
Relationship between Rate of Reaction & [S]

The rate of reaction when the enzyme is saturated with

substrate is the maximum rate of reaction, Vmax.

The relationship between rate of reaction and concentration of

substrate depends on the affinity of the enzyme for its
substrate. This is usually expressed as the Km (Michaelis
constant) of the enzyme, an inverse measure of affinity.

For practical purposes, Km is the concentration of substrate

which permits the enzyme to achieve half Vmax.

An enzyme with a high Km has a low affinity for its substrate,

and requires a greater concentration of substrate to achieve
Vmax.
 Significance of Michaelis Constant(Km) :

• By knowing the Km value of a particular enzyme-substrate system, one can predict whether the cell needs
more enzymes or more substrate to speed up the enzymatic reaction.

• If an enzyme can catalyse a reaction with two similar substrates (e.g., glucose and fructose) in the cell, it will
prefer that substrate for which the enzyme has lower Km value.

• Km value gives an approximate measure of the concentration of substrate of the enzyme in that part of the cell
where reaction is occurring. For instance, those enzymes which catalyse reactions with relatively more
concentrated substrates (such as sucrose), usually have relatively high Km value. On the other hand, the
enzymes that react with substrates which are present in very low concentrations (such as hormones) have
comparatively lower Km values for the substrates.
 Applications of M-M Equation :

• Michaelis–Menten equation is used for modeling drug conversion in the body.

• It is used to characterize the enzymatic rate at different substrate concentrations.

• It is also widely applied to characterize the elimination of chemical (the first-order

kinetics) compounds from the body.

• M–M kinetics have also been applied to a variety of spheres outside of biochemical
reactions, including alveolar clearance of dusts, the richness of species pools, clearance
of blood alcohol, the photosynthesis-irradiance relationship, and bacterial phage
infection.
 Limitations of M-M equation :

M–M equation assumes that one drug molecule is catalyzed sequentially by one enzyme at
a time. However, enzymes may catalyze more than one drug molecule (multiple sites) at a
time, which may be demonstrated in vitro.

In the body, drug may be eliminated by enzymatic reactions (metabolism) to one or more
metabolites and by the excretion of the unchanged drug via the kidney.

The steady-state and rapid equilibrium kinetics do not give detailed information on the
existence of multiple intermediates or on their lifetimes.

When there is a substrate inhibition or activation due to the binding of a second substrate
molecule, the Michaelis–Menten equation does not hold.
THANK YOU