BIOPHARMACEUTICS AND PHARMACOKINETICS

All pharmaceuticals from tablets to state of the art immunotherapy undergo extensive
research and development prior to approval by the FDA.

The physicochemical characteristics of the active pharmaceutical ingredient (API, or

drug substance), the dosage form of the drug, and the route of administration are
critical determinants of the in-vivo performance, safety and efficacy of the product.

Both pharmacist and pharmaceutical scientist must understand these complex

relationships to comprehend the proper use and development of pharmaceuticals.

review your

TERMINOLOGIES USED IN BIOPHAMCEUTICS AND

PHARMACOKINETICS

1. Biopharmaceutics has many definitions, you may use any of these but still it
will sum up to the physicochemical characteristics, biological properties and the
amount the reaches the systemic circulation.
a. It is the study of the relationship of the drug products’ physical and chemical
properties to bioavailability.
b. It is a study of the relationship between physical and chemical and biological
sciences applied to drugs , dosage form and drug action.
c. It is the study of how physicochemical properties of drugs, dosage form, and
routes of administration affect the rate and extent of drug absorption.
d. It is the science that examines the interrelationship of the physicochemical
properties to the drug, dosage form in which the drug is given and the route of
administration on the rate and extent of systemic drug absorption.
e. Describes the extent to which the active ingredient is absorbed from a drug
product and becomes available at the site of action.
f. It is the measurement of the rate and extent of systemic absorption of the
therapeutic drug.

Note: the rate is how fast a drug can be absorbed, distributed and eliminated from
the body and extent is the amount of drug made available in the body.

2. Pharmacokinetics

It is the mathematics of the time course of absorption, distribution, metabolism

and excretion of drugs in the body. After a drug is released from its dosage form,
the drug is absorbed into the surrounding tissues, the body or both (drug
disposition)

3. Clinical Pharmacokinetics –this is the application of pharmacokinetic

methods and drug therapy. It involves a multidisciplinary approach and
individually optimized dosing strategies based on the patient’s disease state, and
 patient’s specific considerations. The study of clinical pharmacokinetics of drugs
in disease states requires input from medical and pharmaceutical research.
4. Pharmacodynamics – this is the study of the biochemical and physiologic
effects of drugs and their mechanism of action.
5. Pharmacogenetics – is the study of the genetic variations that affect drug
response.
6. Drug product- is the finished dosage form that contains the active ingredient in
association with non-drug ingredients (usually inactive, such as excipients) that
make up the vehicle or formulating matrix.
7. Drug delivery system – this includes formulation matrix, its container and the
patient.
8. Bioequivalence is the study of different formulations with compatible
bioavailability when studied in similar conditions.
9. Bioavailability refers to the measurement of the rate and extent of active drug
that reaches the systemic circulation.
10. Toxicokinetics-is the application of to the pharmacokinetic principles to the
design, conduct and interpretation of drug safety evaluation studies and in
validating dose related exposure in animals

INTRODUCTION TO BIOPHARMACEUTICS/ PHARMACOKINETICS

The are several definitions given above for biopharmaceutics which sum up to the
physicochemical factors that influence the rate and amount of drug that is absorbed.
These factors influence:

1. The stability of the drug within the drug product

2. The release of the drug from the drug product
3. The rate of distribution/release of the drug at the absorption site and
4. The systemic absorption of the drug

Absorption
Drug release &dissolution drug in systemic circulation

Excretion and metabolism Drug in tissues

Pharmacologic or clinical effect

Fig. 1 Scheme demonstrating the dynamic relationship between the drug and the drug
product and the pharmacological effect

Biopharmaceutics also considers;

1. The properties of the drug and dosage form in physiologic environment

2. The drugs intended therapeutic use
3. The route of administration
The study of biopharmaceutics is based on fundamental scientific principles and
experimental methodology.

Studies include both:

1. In-vitro methods- are procedures employing test apparatus and equipment

without laboratory animals. The image below shows that tests may be done
using the petri dish method.

Research Gate

2. In-vivo methods are more complex studies involving human subjects or

laboratory animals. The image below shows the use of rats/mice as test
animals.

Creative Animodel

This image shows both an in-vitro and in-vivo methods

These methods must be able to assess:

1. The impact of the physical and chemical properties of drugs

2. The stability of the drug
3. The amount of drug in the body
4. The biologic performance of the drug

HOW CAN WE MEASURE DRUG CONCENTRATION?

Drug concentrations are an important element in determining individual or population
pharmacokinetics. Drug concentrations are measured in biologic samples:

METHODS OF MEASUREMENT
SAMPLING OF BIOLOGIC SPECIMENS

 Invasive methods – include sampling blood, spinal fluid, synovial fluid, tissue
biopsy, or any biologic material that requires parenteral or surgical intervention in
the patient.
 Non-invasive methods- include sampling of urine and saliva, feces, expired air,
or biologic material that can be obtained without parenteral or surgical means

1. Drug concentration in blood, plasma and serum

Measurement of drug concentration levels in the blood, serum or plasma is the
most direct approach to assessing the pharmacokinetics of the drug in the body.
 Whole blood contains cellular elements including red blood cells, white
blood cells, platelets and other proteins such as albumin and globulins.
 Serum of plasma –most commonly used in drug measurement.
To obtain serum the whole blood is allowed to clot and the serum is
collected from the supernatant liquid after centrifugation.
To obtain plasma from the supernatant of the centrifuged whole blood to
which an anticoagulant such as heparin has been added.
 Plasma perfuses all tissues of the body including the cellular elements in
the blood. Changes in drug concentration in the plasma will reflect
changes in tissue drug concentration
2. Drug concentration in tissues
 The measurement of the drug concentration in tissue biopsy material may
be used to ascertain if the drug reached the tissues in the proper
concentration within the tissue.
 Drug concentrations in tissue biopsies may not reflect drug concentration
in other tissues or the drug concentration in all parts of the tissue from
which the biopsy material was removed.
3. Drug concentration in urine
 This is an indirect method to ascertain the bioavailability of a drug.
 The rate and extent of drug excreted in the urine reflects the rate and
extent of systemic circulation
4. Drug in feces
 It may reflect the drug that has not been absorbed after an oral dose, or
may reflect drug that has been expelled from biliary secretion after
systemic absorption.
 Fecal drug excretion is often performed in mass balance studies in which
the investigator attempts to account for the entire dose given to the
patient.
5. Drug concentration in saliva
 Measures free drug that diffuses into the saliva
 Saliva drug levels tend to approximate free drug rather than total plasma
concentration
 The saliva concentrations taken after equilibrium with the plasma drug
concentration generally provide more stable indication of drug levels in
the body.
  The use of saliva drug concentration as a therapeutic indicator should be
used with caution and preferably as a secondary indicator.

DRUG CONCENTRATION IN FORENSICS?

Forensic science is the application of science to personal injury, murder and other
legal proceedings.

Drug measurements in tissues obtained at autopsy or in other bodily fluids such as

saliva, urine and blood may be useful if a suspect or victim has taken an overdose of a
legal medication, has been poisoned or has been using drugs of abuse, such as opiates,
cocaine or marijuana.

SIGNIFICANCE OF PLASMA DRUG CONCENTRATION

1. Monitoring the course of therapy
The intensity of the pharmacologic and toxic effect is often related to the
concentration of the drug at the receptor sites, usually located in the tissue cells.
Because most of the tissue cells are richly perfused with tissue fluids or plasma,
checking the plasma drug level is a responsive method of monitoring the course
of therapy.
2. Monitoring the concentrations of drugs in the blood or plasma ascertains that
the calculated dose actually delivers the plasma level required for therapeutic
effect.
3. Monitoring of plasma drug concentrations allows for the adjustment of drug
dosage in order to individualize and optimize therapeutic drug regimens.

NB: Blood concentration measurement is not always a sure method of predicting

disease states. pharmacodynamics response to the drug may be more important
measure than just plasma drug concentration.
Example: ECG is important to assess in-patients medicated with cardiotonic
drugs such as digoxin; clotting time for prothrombin for an anticoagulant drug;
urine or blood glucose levels for diabetic patients.

ACTIVITY 1

1. Why is plasma or serum drug concentration drug concentration monitored? Why

is this used to determine drug concentration rather than the whole blood?
2. Explain the schematic diagram in Fig. 1
3. What are being assessed by in-vitro and in-vivo studies? Give at least 3 examples
that will involve both studies.
 ACTIVITY 1 (BIOPHARMACEUTICS)
1. Why is plasma or serum drug concentration drug concentration monitored? Why
is this used to determine drug concentration rather than the whole blood?

Measurement of drug concentration levels in the serum or plasma is the

most direct and effective approach to assessing the pharmacokinetics of the
drug in our body. It is monitored to make sure that the amount of medicine that
an individual takes is both safe and effective. Plasma or serum drug
concentration is used rather than the whole blood because whole blood is
generally harder to process and assay than the serum or plasma and it gives a
better result. Also, plasma may be considered a liquid tissue compartment
wherein it perfuses all the tissues in our body including the cellular elements in
the blood. Therefore, changes in the plasma drug concentration reflects changes
in tissue drug concentration.

2. Explain the schematic diagram in Fig. 1

DYNAMIC RELATIONSHIP BETWEEN THE DRUG AND THE DRUG PRODUCT ITS
THE PHARMACOLOGICAL EFFECT

First, the drug product will undergo liberation through disintegration or

dissolution wherein the drug is release and/or dissolved for easier absorption.
After that it will go to the site of absorption to execute the absorption process.
Then when the drug is absorbed it will travel to the systemic circulation through
blood. The drug carried by the blood will now go to the tissues in our body, but
some of it will go back to the systemic circulation and will undergo either
metabolism or excretion because excess drug should be released to avoid
toxicity. Lastly, the drug that went into the tissues will now exert its
pharmacologic or clinical effect into our body.

3. What are being assessed by in-vitro and in-vivo studies? Give at least 3 examples
that will involve both studies.
`

In-vitro study assess the effectiveness of the drug by studying them

outside of the body in which allows a substance to be studied safely without
subjecting humans to possible side effects or toxicity. On the other hand, In-vivo
studies assess how the body as a whole will respond to a particular substance.
Therefore, both studies assess the safeness and effectiveness of a particular
drug or substance.

3 EXAMPLES THAT WILL INVOLVE IN-VITRO AND IN-VIVO STUDIES

 1. In-vitro and In-vivo FERTILIZATION
2. In-vitro and In-vivo HEMOLYSIS
3. In-vitro and In-vivo GENE CLONING

Therapeutic drug monitoring (TDM) is testing that measures the amount of certain

medicines in your blood. It is done to make sure the amount of medicine you are taking
is both safe and effective. Most medicines can be dosed correctly without special
testing.

Therapeutic Drug Monitoring aims to individualize drug therapy and avoid both sub-
therapeutic and toxic plasma drug concentrations. A number of factors influence drug
concentration and a single sample will only reflect the concentration at the sampling
time.
Measuring the plasma concentration of a drug allows the doctor to track the dosage
to the individual patient and to obtain the maximum therapeutic effect with minimal risk
of toxicity. 

Blood is composed of plasma and red blood cells (RBCs). Serum is the fluid obtained
from blood after it is allowed to clot. Serum and plasma do not contain identical proteins.
RBCs may be considered a cellular component of the body in which the drug
concentration in the serum or plasma is in equilibrium, in the same way as with the
other tissues in the body. Whole blood samples are generally harder to process and
assay than serum or plasma samples. Plasma may be considered a liquid tissue
compartment in which the drug in the plasma fluid equilibrates with drug in the tissues
and cellular components.

3.

As a Latin term that means the “whole body,” in vivo refers to tests that are performed in a live
environment of a whole body—usually on live animals such as murine models. This step
supports drug safety testing before the launch of human clinical study—but there are
challenges, too.

In vivo is Latin for “within the living.” It refers to work that’s performed in a whole, living
organism.

A test performed in vitro ("in the glass") means that it is done outside of a living

organism and it usually involves isolated tissues, organs or cells.

In vitro

In vitro methods used in a laboratory can often include things like studying
bacterial, animal, or human cells in culture. Although this can provide a
controlled environment for an experiment, it occurs outside of a living
organism and results must be considered carefully.

EXAMPLES:

Fertilization

You’ve likely heard of in vitro fertilization (IVF). But what exactly does that mean?

IVF is a type of treatment for infertility. In IVF, one or more eggs are removed from
an ovary. The egg is then fertilized in a laboratory and implanted back into the uterus.

Because fertilization occurs within a laboratory environment and not within the body (in
vivo), the procedure is referred to as in vitro fertilization.

Antibiotic sensitivity

Antibiotics are medications that work to treat bacterial infections. They do this by

disrupting the bacteria’s ability to grow or thrive.

There are many types, or classes, of antibiotics and some bacteria are more sensitive to
some classes than others. Additionally, bacteria can evolve to be resistant against
antibiotics.

Although bacterial infections occur on or in our bodies, antibiotic sensitivity testing often

occurs within a laboratory setting (in vitro).
In vivo

When a study is performed in vivo, it can include things like performing

experiments in an animal model, or in a clinical trial in the case of humans. In
this case, the work is taking place inside a living organism.

DEFINITIONS OF IN VIVO AND IN VITRO

The terms “in vitro” and “in vivo” are used quite frequently in scientific articles and
research. Even if you don’t work in the healthcare or research field, you’ve probably
come across them before. “In vivo” and “in vitro” are basically two different types of
methods that can be used to describe experiments used in clinical trials, scientific
studies, and certain medical procedures.

In vivo

In Latin, in vivo means “within the living,” and it refers to experiments that are carried out
using a whole, living organism rather than dead or partial samples (in-vitro
environments). Two of the main examples of in vivo experiments are clinical trials and
animal testing.

In some cases, in vivo experiments can help researchers determine the effects of a
procedure or medication on living beings. In microbiology, the term “in vivo” is also used
to refer to experiments on live, isolated cells instead of a whole organism.

In vitro

In Latin, in vitro means “in glass.” In vitro studies use isolated components of an

organism that have been separated from their usual surroundings. Looking at only
specific components instead of the whole organism can make experiments easier for
researchers and allows for more detailed analyses.

In vitro tests have allowed researchers to test out many new different drugs at once.

These experiments are sometimes casually known as “test tube experiments.” They
provide a safe way for scientists to perform early tests on new procedures and
treatments without putting live subjects at risk. In vitro tests have allowed researchers
to test out many new different drugs at once. Only the ones that appear to be safe and
effective go on to the clinical trial stage.

In vitro techniques are also used in certain medical procedures, particularly assisted
fertility treatments. These treatments have allowed many couples who previously
couldn’t have conceived to get pregnant and start their own families. 

In vivo vs. in vitro: differences and similarities

In vivo and in vitro are different techniques that can be used for different things, but
there are also certain similarities between them. Here are some of the differences and
similarities between these two testing techniques:
  In vivo testing is more expensive than in vitro, and it takes longer to get results.
 Most biological experiments are carried out using in vitro techniques; however, in vivo
testing produces more specific and detailed results because it simulates the biological
conditions found in a live subject.
 In vivo testing has to adhere to more regulations because it uses live subjects.
 Both types of tests are used to develop new treatments that have advanced science and
medicine.

How are these experiment types used in clinical studies?

When a new drug is being developed, initial testing is carried out in vitro. Researchers
add the drug to a Petri dish that contains cells and closely monitor its effects. For
example, researchers may add a new chemotherapy drug to a dish containing cancer
cells. However, isolated cells in a Petri dish don’t necessarily react the same way that
cells inside a living being do. If in vitro research determines that the drug is potentially
effective and safe, it advances to in vivo tests.

Animal testing is another form of in vivo experimentation. If in vitro tests are successful,
many new drugs are tested on animals, such as laboratory rats, to see how the drug
acts on live subjects. However, many drugs work differently on human beings than they
do on laboratory animals, which is why animal testing isn’t enough to ensure that a drug
is completely safe.

If in vitro research determines that the drug is potentially effective and safe, it advances
to in vivo tests.
After animal testing, new medications undergo further in vivo testing during clinical
trials to determine their effects on human beings before they can be approved for
general use. In many cases, researchers compare the effects of new drugs against a
placebo. It’s important to note that clinical trials are voluntary, and patients can
withdraw from them whenever they want. 

In vitro fertilization, or IVF, is a procedure in which a woman’s mature eggs are retrieved

from her ovaries, and then sperm — coming from her partner or a donor — is used to
fertilize the egg. The resulting fertilized eggs or embryos are implanted into the
woman’s uterus for a pregnancy to develop.

In vitro fertilization is a procedure in which a woman’s mature eggs are retrieved from her
ovaries, and then sperm — coming from her partner or a donor — is used to fertilize the
egg.
There are several reasons why people choose IVF. Among others, some women want to
become mothers at an older age, and some have fertility issues due to a medical
condition.

In vitro testing is also commonly used for antibiotic sensitivity testing, which allows
doctors to determine exactly which antibiotic they should use to effectively treat a
patient’s infectious disease. This can also decrease the risk of creating multi-resistant
bacteria when different antibiotics are used to treat a disease.

Biomedical researchers constantly use both in vivo and in vitro techniques to develop
new drugs, diagnose conditions, and create new treatments.
Some biopsies represent a slightly different type of testing, which is called “ex vivo.”
Pathologists can run tests on live cultured cells that come from a biopsy sample to rule
out different conditionsand provide a diagnosis. In ex vivo tests, researchers have
removed the live cells from the organism and added them to an artificial environment,
trying as much as possible to keep natural conditions the same.