Biopharmaceutical
analysis
By
Dr. Aliaa Magdy
Radwan
2022/2023
Drug discovery and design
• Drug (pharmaceutical) is defined as a chemical substance that is used
to treat, cure or prevent diseases in humans and animals.
• Drug discovery is the process through which potential new therapeutic
entities are identified, using a combination of computational,
experimental, and clinical models.
1. The synthesis of the drug .
2. The method of administration
3. The development of tests and procedures to establish how it operates in the body .
4. A safety assessment.
• There is no drug which is completely
safe, including non-prescription drugs
such as aspirin and paracetamol.
• Overdoses of paracetamol can cause
coma and death.
• Aspirin can cause gastric irritation and
bleeding in some people if taken for
long time or in big doses.
• These unwanted effects are commonly
referred to as side effects.
• The antihistaminic promethazine is
licenced for the treatment of hayfever
but also induces drowsiness, which may
aid sleep
• Tachyphylaxis (drug resistance or tolerance) is a medical term that defined as a
rapid appearance of progressive decrease in response to a given dose after
repetitive administration of a pharmacologically or physiologically active
substances such as antibiotics.
• There are different reasons that may be responsible for drug resistance:
1. Increase in the rate of production of an enzyme that metabolizes the drug.
For example, the effectiveness of barbiturates often decreases with repeated
use
2. Down-regulation of receptors
3. The appearance of a significantly high proportion of drug resistant strains of
microorganisms.
History of drug discovery
• In the ancient times, folk medicines were used (mainly derived from plant
products, animals, or mineral sources).
• In the middle ages (400-1500), different diseases are spread out in Europe
such as bubonic plague, leprosy, smallpox, tuberculosis, and scabies.
• In the late 18th century, there were many advances made in medicine, laid
the foundation for scientific thoughts in medicinal preparations and
medical treatments. (Edward Jenner ’ s Smallpox Vaccine)
• In the late 19th century, the drug discovery and development started to
follow scientific techniques. More and more drugs were discovered, tested,
and synthesized in large scale.
Drug discovery
• In 20th century, the beginning of modern pharmaceutical industry and
the development of synthetic drugs (Paul Ehrlich who produced the
antiprotozoal arsphenamine (Salvarsan)
• First rational drug design, wanted to maximize toxicity to pathogen and
minimize toxicity to human (Therapeutic index)
• Therapeutic index (TI)= LD50/ED50
• LD50: lethal dose that required to kill 50% of the tested animals
• ED50: the dose that required to produce therapeutic effect in 50% of
the tested animals
Structure–activity relationship (SAR)
• SAR is Ehrlich’s approach that is used nowadays in drug discovery
research to predict the biological activity of structurally related
compounds.
• QSAR (quantitative structure activity relationship) is a computational
model that used to predict the physicochemical and biological
properties of novel molecules prior to their synthesis.
Receptive substances
• was formulated by John Langley in 1905 to explain the theory of drug action.
• He proposed that receptive substances present in the body.
• Drugs can be classified into two types: Agonist and Antagonist.
• These receptive substances could receive stimulating compound (Agonist) that
cause biological response or non-stimulating compound (Antagonist) that
prevent biological response.
• This theory enabled the chemists to place groups in the molecular
structure of the drug to enhance its absorption, distribution,
metabolism, and excretion (ex: sulphonic acid group).
Local anesthetic anti-rhythmic
Cholinesterase inhibitors
Lecture 2
Classification of drugs
• The drugs can be classified according to:
1) Chemical structure
2) Pharmacological action
3) The site of target system (Physiological classification)
1) Chemical structure
• The drugs can be classified depending on their carbon skeleton into
steroids, peptides, and penicillins.
Sex hormone Diuretic agent antibacterial agent
2) Pharmacological action
• The drugs can be classified depending on their pharmacodynamics
behavior (How a drug affects or influences the cells of an organism).
• Pharmacodynamics: is the study of the biochemical and physiologic
effects of pharmaceutical drugs
• Analgesics (pain killers), anti-inflammatory, diuretics (water pills),
vasodilators, and antimicrobial (antibiotics such as penicillin)
3) Physiological classification
• According to WHO, the drugs can be classified based on the target site they act
on.
1) Agents acting on the central nervous system (CNS) : such as that used for
nervous disorders (epilepsy and pain)
2) Pharmacodynamic agents: include vasodilators and antiallergy agents
3) Chemotherapeutic agents: antibiotics and fungicides
4) Miscellaneous agents: such as hormones and drugs that act on endocrine
functions
5) Prodrugs: pharmacologically inert but converted to the active form at or near
their target site
Used to treat Parkinson’s syndrome
Leads and analogues
• The drug discovery and development process begins with the discovery of
a lead compound that exhibits the desired biological activity.
• Structure-activity relationship (SAR) was then studies by organic chemists
• Analogues are derivatives of lead compound with optimized pharmacokinetic
or pharmacodynamic properties
Lead compound Analog
• Leads and analogues should have some desirable properties such as:
• Bioavailability
• Solubility
• Structure
• Stability
1) Bioavailability (F)
• is a measure of the rate and fraction of the initial dose of a drug that successfully
reach the systemic circulation (measures the actual amount of drug that
reaches the systemic circulation following oral dosing)
• It is a part of pharmacokinetics paradigm that studying the movement of drug
through the body (ABCD).
• The route of administration (ROA) and the dose of a drug have a significant
impact on both the rate and extent of bioavailability
• The dose of a drug is indirectly proportional to its bioavailability
Bioavailability Dosage of drug
To reach the minimum effective concentration
Bioavailability assessment
• In 1997, Lipinski et al. proposed a set of four rules that would predict
whether a molecule was likely to be orally bioavailable as follow:
1) MW ≤ 500 Da
2) Hydrogen bond acceptors(HBAs such as N,O) ≤ 10
3) Hydrogen bond donors (HBD such as NH,OH) ≤ 5
4) Calculated LogP (cLogP) ≤ 5 and ≥ 0
• Lipophilicity (LogP) is one of the most important properties of all these
rules, defined as the partition coefficient ratio of a compound between the
hydrophobic and hydrophilic phases
• A partition coefficient (P) is the ratio of concentrations of a compound in a
mixture of two immiscible solvents at equilibrium (Octanol/water).
2) Solubility
• The drugs should have some extent in both lipid and water.
Lipophilic (hydrophobic) compounds hydrophilic (lipophobic) compounds
dissolve in lipid solvents do not dissolve in lipids but dissolve in water
contain large numbers of non-polar groups, contain polar groups such as acid, amine and
such as benzene rings hydroxy functional groups
Poor water solubility prevent the Should have degree of lipid solubility to pass
development of drugs such as Taxol through membrane
It is desirable that leads and analogues have a balance between their water
solubility and their lipophilicity
3) Structure
• A major consideration in the selection of leads and analogues is their
stereochemistry
• The interaction between drugs and receptors or enzyme may be reversible or
irreversible.
• Reversible: The drug or analogue forms week, electrostatic bonds, such as
hydrogen bonds and van der Waals’ forces, with the receptor (ex:local
anaesthetic benzocaine)
• Irreversible: drugs and analogues act by forming strong covalent bonds with
the receptor (ex: melphalan and omeprazole).
• Omeprazole used for the treatment of gastric ulcers by inhibiting the secretion
of gastric acid.
4) Stability
• Drug stability refers to the extent to which a pharmaceutical substance is
able to keep its therapeutic properties throughout its storage period or
shelf life
• It can be broadly divided into two main areas: stability after
administration and shelf-life
• Stability after administration: the drug must be
stable enough to reach its target site in sufficient
concentration and must not be metabolized too
quickly.
Three strategies are commonly used for improving a
drug’s stability
1. Modifying its structure.
prepare a more stable analogue with the same pharmacological activity
For example: Pilocarpine which is used to control glaucoma
Cyclodextrins
• In the pharmaceutical industry, cyclodextrins have mainly been used as
complexing agents
• They are bottomless flower-pot-shaped cylindrical oligosaccharides consisting
of about 6–8 glucose units
• The formation of these complexes may improve the water solubility,
bioavailability and pharmacological action and reduce the side effects of some
drugs
• For example, complexing with hydroxypropyl-β-cyclodextrin is used to improve
both the stability and solubility of thalidomide
2. Administering the drug as a more stable prodrug
• Prodrug formation can also be used to improve drug stability (For example,
cyclophosphamide)
• Its drug stability in the gastrointestinal tract can be improved using enteric
coatings, which dissolve only when the drug reaches the small intestine.
• once a drug has carried out its function it needs to be removed from the body
3. Using a suitable dosage form
Shelf-life
• Shelf-life is the time taken for a drug’s pharmacological activity to decline to an
unacceptable level.
• Shelf-life deterioration can be occurred due to :
➢Microbial degradation
It can be avoided by preparing the drug in sterile conditions and using
antimicrobial vehicles.
➢Chemical interactions
It is usually occurred by heat, light, atmospheric oxidation, and hydrolysis by
atmospheric moisture.
It can be avoided by correct storage of drug, light-proof containers, and air-tight
lids.
Sources of drugs
and drug action
LECTURE3
Sources of leads and drugs
1) Ethnopharmaceutical sources
• By studying and screening the ancient and traditional folk medicine
(Ethnopharmacology).
• For example, the antimalarial quinine from cinchona bark, the cardiac
stimulants from foxgloves and the antidepressant reserpine isolated
from Rauwolfia serpentina.
• Common foxglove leaves contain about 30 different cardioactive
compounds.
R1=H is called digitoxigenin
• Digitoxin is a trisaccharide derivative of digitoxigenin, is the only Digitalis purpurea, common foxglove
compound to be used clinically to treat congestive heart failure
2) Plant sources
• All parts of plant, from roots to seeds and flowers can act as a
source of a lead compound.
• The collecting of plant samples must be carried out with due
consideration of its environmental impact.
• It is essential that a full botanical record of the plant is made if it
does not already exist. This record contain a description and
pictures of the plant and any related species, and its growing
conditions.
• A detailed record:
➢ The date of the collection of the samples
➢ The method used for its collection,
➢ its harvest site storage
➢ method of preparation for onward transportation to the
investigating laboratory
Botanical record
• Plant samples are normally extracted and put
through screening programs.
• One of the most common problem in drug discovery
from plant source is the extraction, purification and
assessment of the pharmacological activity of an
active compound which may cause ecological
problem (for example, the anticancer drug Taxol).
• Examples of some of the drugs in clinical use
obtained from plants. Taxol and vincristine are
anticancer agents isolated from Taxus breifolia and
Vinca rosea Linn, respectively. Pilocarpine is used to
treat glaucoma and is obtained from Pilocarpus
jaborandi Holmes Rutaceae. Morphine, which is used
as an analgesic, is isolated from the opium poppy.
Pacific Yew (Taxus breifolia) tree
3) Marine sources
• These marine sources including, marine microorganisms, fungi, shellfish, and sponges.
• In the last 40 years, there are large number of active compounds and drugs have been
produced from these sources.
• These compounds have different biological activities and are an important source of new
lead compounds and drugs.
• Examples of active compounds isolated from marine sources:
➢ Avarol is an immunodeficiency virus inhibitor and is extracted from the sponge Disidea
avara
➢ The antibiotic cephalosporin C was isolated from the fungus Cephalosporin acremonium.
➢ Tetrodotoxin and saxitoxin exhibit local anaesthetic activity.
➢ Ara-A is an FDA - approved antiviral isolated from the sponge Tethya crypta
4) Microorganisms
• In 1877, Louis Pasteur was the first one who observed the inhibitory effect of
microorganism and showed that they can inhibit the growth of anthrax bacilli in urine.
• In 1920, Fleming demonstrated that Penicillin notatum inhibited staphylococcus cultures
that resulted in the isolation of penicillin by Chain and Florey in 1940.
• In 1941, Dubos isolated the antibiotic gramicidin from Bacillus brevis.
• In 1944, they discovered and isolated streptomycin antibiotic from actinomycete
Streptomyces griseus.
• To date, there are several thousand active compounds have been discovered from
microorganisms, for example the antibiotic chloramphenicol (Streptomyces venezuelae),
the immunosuppressant cyclosporin A
5) Animal sources
• There are different drugs have been extracted and
isolated from animal sources and used to treat
different diseases.
• For example:
➢ Adrenaline hormone
➢ Insulin was first commercially produced from bovine
and porcine sources then in the last 20 century it was
produced from bacteria using recombinant genetic
engineering.
Drug action
• Drug action is believed to be due to the interaction of
that drug with enzymes, receptors or other molecules
found in the body, resulting in change in or inhibit the
biological activity of these molecules.
• Factors affecting the concentration of drug in
biological system:
➢ The pharmacokinetic phase
➢ The pharmacodynamic phase of drug action.
The pharmacokinetic phase(ADME)
• concerns the study of the parameters that control the journey of the
drug from its point of administration to its point of action .
• This include absorption, distribution, metabolism, and excretion of
the drug.
Absorption of drug
Absorption is defined as the passage of the drug from its site of
administration into the general circulatory system after enteral
administration (oral administration).
The absorption of drugs through membranes and tissue barriers can
occur by a number of different mechanisms.
Neutral molecules are more readily absorbed through membranes than
charged species.
Factors affecting drug absorption
1) The drug solubility and the balance between hydrophilic and
hydrophobic groups.
• Drugs that are too polar are absorbed by paracellular diffusion
(passive transport) while less polar or non-polar drugs are
absorbed by transcellular diffusion (active transport).
Factors affecting drug absorption
2) the surface area of the region of tissue over which the absorption is occurring and the time the drug spends in contact with that
region.
for example: Ionization of orally administered aspirin is suppressed in the stomach by acids produced. As a result, it is absorbed in
this uncharged form through the stomach lining into the blood stream in significant quantities.
On the other hand, aspirin will be almost fully ionized in the small intestine. Consequently, aspirin should not be readily absorbed in
this region of the GI tract. However, the very large surface area of the small intestine (300 m2 ) together with the time spent in this
region ( 6 hours) results in aspirin being absorbed in significant quantities in this region of the GI tract.
3) the pH of the medium from which absorption occurs
4) the drug’s dosage form
5) the drug’s particle size
6) rate of dissolution
Drug dosage form
• The form in which a medicine is administered is known as its dosage form. It can be
subdivided into:
• Liquid: solutions, suspensions, and emulsions.
• Semisolid: Creams, ointments and gels.
• Solid: tablets, capsules, and suppositories.
• These dosage forms normally consist of the active constituent and other
ingredients (excipients).
• Excipient is an inactive substance that formulated alongside the active ingredient
of drug in a drug delivery system in order to aid in the manufacturing process and
enhance stability, bioavailability, safety, effectiveness and delivery of the drug
including, fillers, binders, preservative or antioxidant.
• A change in the nature of the excipients can significantly affect the release of the
active ingredient from the dosage form. For example, the anticonvulsant phenytoin.
compression and before releasing into the packing
