Pharmaceutical Biotechnology. INTRODUCTION TO PHARMACEUTICAL BIOTECHNOLOGY & SOURCES OF BIO-PHARMACEUTICAL PRODUCTS

1. Pharmaceutical Biotechnology (Biot 3112)
For Third Year Biotechnology Students.
Instructor: - Temam 2G (M.Sc. In Biotechnology)
WACHEMO UNIVERSITY 2022
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2. UNIT ONE
Historical overview of Pharmaceutical biotechnology,
pharmaceutical products & Sources of pharmaceutical
products
Objective: at the end of this class you will understand:-
What is Pharmaceutical Biotechnology
What is Pharmacology (Science of drug)
AIM of pharmaceutical biotechnology
History of Pharmaceutical industry
Biopharmaceuticals
Pharmaceutical biotechnological product
Opportunities for Pharmaceutical Development
Biopharmaceuticals approved in USA &EU
Biopharmaceutical Companies
Requirements of successful development of biopharmaceuticals
Therapeutic and Pharmaceutical applications of Biotechnology

3. Pharmaceutical Biotechnology
 Pharmaceutical biotechnology is combination of two
branches:-
Pharmaceutical science and
Biotechnology.
 Pharmaceutical Science: - a science study about
formulation, compounding & administration of drugs
 Biotechnology: - The application of:-
Living organisms
Part of living organisms, or
their derivatives :- in making or modifying products
Pharmaceutical biotechnology:- study of production,
manufacturing and registration of biopharmaceuticals.
 biopharmaceuticals:-biological drugs
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4. Pharmacology (Science of drug)
 It is study about drugs, their preparation and properties
and uses and effects in medicine
 Drugs:- a substance that is used as a medicine (narcotic).
 Medicine:- Something that treats, prevents & relief symptoms of
diseases or
 branch of medical sciences that study about nonsurgical techniques
 Pharmaceutical biotechnology use microorganisms or
hybrids of tumor cells and leukocytes to create /produce:-
 new, safe and more effective pharmaceuticals
 pharmaceuticals identical to conventionally made and more cost-
effective than conventionally made.
Ex. Productions of recombinant human insulin in 1982
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5. AIM of pharmaceutical biotechnology
 It is design, and production of drugs that are:-
Adapted to each persons genetic make up
Give the maximum therapeutic effect.
Role biotechnology to success aim of pharmaceutical biotechnology
Production of genetically modified organisms used for
pharmaceutical industries.
Principles of biotechnology is used to develop drugs
 Majority of therapeutic drugs are bio-formulations
Examples:
antibodies,
nucleic acid products and
vaccines 5

6.  Stages of bio-formulations drug developing
Understanding principles of health and disease
Governing function of related biomolecules
Synthesis and purification of molecules
 Determining product shelf life, stability, toxicity and
immunogenicity
Drug delivery systems; patenting and clinical trials.
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7. History of the pharmaceutical industry
The pharmaceutical industry is barely 80 years old.
From very beginnings, it has grown rapidly, reaching an
estimated value of US$100 billion by the mid 1980s.
Its current value is likely double
There are more than 10,000 pharmaceutical
companies in existence in today world
From them only about 100 of these can claim to be of
true international significance.
These companies manufacture more than 5000
individual pharmaceutical substances used medicine.
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8. The history of pharmaceutical biotechnology
includes Alexander Fleming’s discovery of
penicillin in a common mold, in 1928
The subsequent development prompted by World
War II injuries of large-scale manufacturing
methods to grow the organism in tanks of broth.
Pharmaceutical biotechnology has since changed
enormously.
Some pharmaceuticals were traditionally obtained
by direct extraction from biological source material.
Many of the protein-based pharmaceuticals are
produced by genetic engineering
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9. Common Terms used pharmaceutical literature
 Terms of pharmaceutical literature:
 biologic
 biopharmaceutical
 products of pharmaceutical biotechnology or
 biotechnology medicines
 These terms are sometimes used interchangeably
 Ex. biologic’ refers medicinal products derived from
biological such as blood, vaccines & toxins
 They have different meaning in different situation.
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10. Biopharmaceuticals
The term Biopharmaceutical was first used in
1980s to describe:-
Therapeutic proteins produced by genetic
engineering or hybridoma technology and
Nucleic acid and whole-cell-based products

11. Biopharmaceuticals

13. PHARMACEUTICAL BIOTECHNOLOGICAL
PRODUCT
 Currently there are many pharmaceutical
biotechnological product that are used for therapeutic
purpose.
 These pharmaceutical biotechnological products are:-
 Antibodies
Recombinant DNA Products.
Cytokines and growth factors
Hormonal products
Recombinant blood products
Therapeutic enzymes
Vaccines and adjuvants 13

14. Biopharmaceuticals approved in USA &EU Over last 25yr
 Recombinant products like:-
1.Blood factors, e.g. Factor VIII :-Treat Hemophilia A
2.Thrombolytic & anticoagulants:-treat myocardial infarction(heart attack
3.Hormones, e.g. insulin, HGH:-
 Treats diabetes mellitus, growth disturbances in children
4.Growth factors, e.g. erythropoietin :-Treats Anemia.
5.Interferons and Interleukins:- e.g. Interferon-α & -β
 Treats Hepatitis B, C and various cancers
6. Vaccines:- e.g. Hepatitis B :-Treat Hepatitis B
7. Monoclonal antibodies:- e.g. Herceptin, ProtaScint
 Treats breast cancer, prostate adenocarcinoma
8. Enzymes, e.g. Myozyme :-Treat Pompe disease
9. Nucleic acid products, e.g. Macugen:-Treats macular degeneration
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15. Biopharmaceutical Companies
 The first known biopharmaceutical Companies in world
are :
1. Amgen
2. Biogen and
3. Genentech
 They are pioneering of biopharmaceutical companies
1. AMGen:- (Applied Molecular Genetics) is founded in 1980s
 Focus on developing novel therapeutics for:-
Oncology,
Inflammation,
Bone disease,
Neurology,
Metabolism and nephrology. 15

16. 2.Biogen:-
 Founded in Geneva, Switzerland, in 1978
 It developed and markets interferon based product
Avonex (intf beta-1a treat sclerosis)
3.Genentech:- founded in 1976 by Herbert Boyer and
Robert Swanson.
 It has 10 protein-based products on the market.
 These include:-
hGHs (Nutropin),
Herceptin’ and ‘Rituxan (antibody- based products),
thrombolytic agents (Activase’ and ‘TNKase)
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17. Role of Recombinant DNA and Molecular Genetics
Advance in Biopharmaceutical
 Recombinant DNA and Molecular Genetics Advance in
1970s and 1980s
 It result in advancing:
 Biopharmaceutical class of drugs
 Improved methods for finding interesting molecules.
 This advent help scientists as:-
predict alter protein’s sequence and produce altered
protein in quantity
Allowed rationalize relationships of structure and
function in drug design.
Allowed production of recombinant proteins for
drugs
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18. Opportunities for Pharmaceutical Development
 Opportunities of Pharmaceutical development are
developments of:-
New technologies for investigating complex biological
systems
New technologies for measuring drug effects
New technologies for predicting outcomes
Effective Integrating New Technologies
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19. Potential of biopharmaceutical for Advances of
Medicines
The Potential of biopharmaceutical for Advances of Medicines is:-
i. Unmet Medical Needs
 Treatments for known diseases that currently lack treatments
 Treatments for diseases not yet recognized
ii. Drug Efficacy:-More reliable patient response to therapies
iii. Drug Safety:- Fewer side effects
Challenges for Pharmaceutical Innovation from Current Advances
a. Effective acquisition and integration of technological advances
b. Conversion of data from genomics, proteomics and other high-
throughput data-gathering technologies into medically relevant
knowledge
c. Successful application of that knowledge toward improved
productivity in drug development

20. Significance of Biopharmaceuticals / Recombinant
Proteins
i. It overcomes the problem of source availability:
 Many proteins of therapeutic potential are produced
naturally in the body in minute quantities.
 Examples:- interferon's, interleukins and colony-stimulating
factors.
 This rendered impractical their direct extraction from
native source material in quantities sufficient to meet likely
clinical demand.
 Recombinant production allows the manufacture of any
protein in whatever quantity it is required.
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21. ii. It overcomes problems of product safety.
Direct extraction of product from some native
biological sources in the past has led to the
unwitting transmission of disease.
Examples;- the transmission of blood-borne
pathogens such as: hepatitis B and C and human
immunodeficiency virus (HIV) via infected blood
products and the transmission of Creutzfeldt–
Jakob disease to persons receiving human growth
hormone (GH) preparations derived from human
pituitaries.
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22. iii. It provides an alternative to direct extraction from
inappropriate/dangerous source material.
A number of therapeutic proteins have been
traditionally extracted from human urine.
Example, Follicle stimulating hormone (FSH), the
fertility hormone is obtained from the urine of
postmenopausal women, and a related hormone,
human chorionic gonadotrophin (hCG), is extracted
from the urine of pregnant women.
Urine is not considered a particularly desirable source
of pharmaceutical products.
Other potential biopharmaceuticals are produced
naturally in downright dangerous sources.
recombinant production in less dangerous organisms,
such as Escherichia coli or Saccharomycese cerevisiae,
would be considered preferable by most. 22

23. Requirements of successful development of
biopharmaceuticals
The successful development of biopharmaceuticals
requires:
Advanced biochemical/biomedical research to identify
and characterize the native compounds
Skilled molecular biology and cloning technology to
identify the relevant gene sequences and insert them
into a production mammalian or microbiological host
Bioprocess technology to grow the organisms and to
isolate, concentrate and purify the chosen compounds
Clinical and marketing expertise.
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24. Therapeutic and Pharmaceutical applications of
Biotechnology
 Biotechnology helps the pharmaceutical industry to
develop new products, new processes, methods and
services and to improve existing ones
 With biotechnology techniques, drugs are developed
using microorganisms with no use of chemicals or
synthetic materials. These drugs are usually sourced from
large molecules of proteins. Thus they attack the hidden
mechanisms of the disease and destroy them without any
side effects
 The major Therapeutic and Pharmaceutical applications
of Biotechnology is Gene Therapy, Pharmaco-genomics,
Genetic Testing & manufacture of vaccines

25. 1. Gene Therapy:-
 used to diagnosing diseases like Parkinson’s and cancer
2.Pharmaco-genomics:-
 it’s a combination of genomics and pharmaceuticals
 It is genetically modified technique used for learning genetic
information of a person
 It analyses the body’s response to drugs.
3. Genetic Testing
 It used to find genetic diseases in parents, sex and carrier
screening
 It used DNA probes that have some sequence identical to the
mutated sequence
4. manufacture of vaccines
 It is genetically engineering and cell culture enable an effective,
quick and economical development of vaccines.
 Recombinant DNA technology enables antigen of a certain
pathogen to be produced in a host cell which is relatively non
pathogenic

26. Pharmaceutical
Biotechnology
CHAPTER 2
Sources of Biopharmaceuticals products
Wachemo university
By:- Temam 2g
2021

27. UNIT TWO
SOURCES OF BIOPHARMACEUTICALS PRODUCTS
 Objectives: at the end of this class you will understand:-
Sources of Biopharmaceuticals
 Heterologous protein production
Use of E. coli in recombinant biopharmaceutical production
Advantages of using E. coli in recombinant production
Disadvantages of using E. coli in recombinant production
Fungal &Yeast cell in recombinant biopharmaceutical
production
Disadvantages of using Yeast cell-based system
biopharmaceutical production
Use of Animal cell culture system in Biopharmaceuticals
Advantages & Disadvantages of Animal cell-based culture
systems biopharmaceutical production

28. Sources of Biopharmaceuticals
 Biopharmaceutical, also known as a biologic(al) medical
product, or biologic,
 Biopharmaceutical is any pharmaceutical drug product
manufactured in, extracted from, or semisynthesized from
biological sources
 Biopharmaceuticals may be produced from:-
Microbial cells (e.g., recombinant E. coli or yeast
cultures)
Mammalian cell lines (mammalian Cell culture) and
Plant cell cultures (Plant tissue culture)
 semisynthesized differ from Total synthesis that are
complex molecule that completely chemically synthesized
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29. Some pharmaceutical product from chemical sources
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30. Biopharmaceuticals produced from Biological sources
 It the drugs produced by genetic engineering
 They are recombinant proteins produced either:
In recombinant E. coli or
Mammalian cell lines or
Plant cell cultures (Plant tissue culture)
 It produced by introducing cDNA coding for protein of
interest into producer cell.
• Ex. E. coli K12
 Microorganisms used for such activity must be cultured
i. in large quantities,
ii. inexpensively and
iii. In short time by standard fermentation
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31. Pharmaceuticals product from Biological sources
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32. Heterologous protein production
 It expression of recombinant proteins in cells in which
they do not naturally occur
 Used in production of biological drug
Use of E. coli in recombinant biopharmaceutical production
 E. coli is a popular expression platform for the production of
recombinant proteins used in therapeutics
 The first biopharmaceutical produced by genetic engineering that
gain marketing approval in 1982 was recombinant human insulin
(tradename ‘Humulin’) produced in E. coli.
 Example of more recently approved biopharmaceutical that is
produced in E. coli is Kepivance, a recombinant keratinocyte
growth factor used to treat oral mucositis
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33. Advantages of using E. coli in recombinant production
is:-
long served for studies
Highly of expression of heterologous proteins
(expression of the recombinant protein reach up to
30% total cellular protein)
High-expression promoters
Its cells grow rapidly
Use inexpensive media
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34. Drawbacks of E. coli as a biopharmaceutical producer.
 Drawbacks of using E. coli as a biopharmaceutical
producer include:-
Heterologous proteins accumulate intracellularly.
Inability to undertake post-translational modifications
(particularly glycosylation) of proteins. Glycosylation is
the process of adding glycosyl radicals to a protein to
form a glycoprotein. Glcosyl is monovalent radical
derived from a cyclic form of glucose by removing of
hemiacetal hydroxy group.
Presence of lipopolysaccharide (LPS) on its surface.
The vast bulk of proteins synthesized naturally by E.
coli homologous proteins are intracellular.
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35. Few are exported to the periplasmic space or released
as true extracellular proteins.
Heterologous proteins expressed in E. coli thus
invariably accumulate in the cell cytoplasm
Intracellular protein production complicates
downstream processing (relative to extracellular
production) as additional primary processing steps are
required, i.e. cellular homogenization with subsequent
removal of cell debris by centrifugation or filtration
More extensive chromatographic purification is
required in order to separate the protein of interest;
from the several thousand additional homologous
proteins produced by the E. coli cells.
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36.  An additional complication of high-level intracellular
heterologous protein expression is inclusion body
formation. Inclusion bodies (refractile bodies) are
insoluble aggregates of partially folded heterologous
product. Because of their dense nature, they are easily
observed by dark-field microscopy. Presumably, when
expressed at high levels, heterologous proteins overload
the normal cellular protein-folding mechanisms.
 However, the formation of inclusion bodies displays one
processing advantage: this advantage is facilitates the
achievement of a significant degree of subsequent
purification by a single centrifugation step. Because of
their high density, inclusion bodies sediment even more
rapidly than cell debris. Low speed centrifugation thus
facilitates the easy and selective collection of inclusion
bodies directly after cellular homogenization.
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37.  After collection, inclusion bodies are generally incubated with
strong denaturants such as detergents, solvents or urea. This
promotes complete solubilization of the inclusion body i.e.
complete denaturation of the proteins therein. The denaturant
is then removed by techniques such as dialysis or filtration
Shortly of using E. coli in recombinant biopharmaceutical
production has the following drawbacks.
Heterologous proteins accumulate intracellularly
Glycosylation of proteins
Glycosylation:-Inability to undertake post-translational
modifications or
adding glycosyl radicals to a protein to form a glycoprotein.
presence of lipopolysaccharide (LPS) on its surface.
inclusion body formation

38. Fungal production systems
 It produce heterologous protein in industrial enzymes
Ex. α-amylase and glucoamylase.
 It express high various extracellular proteins
 It carry out post-translational modifications.
Yeast cell-based system
 It used for production of several recombinant
biopharmaceuticals approved for medical use
Disadvantages of Yeast cell-based system
Their glycosylation pattern varies from native
glycoprotein
Express <5% of heterologous proteins of cellular
protein.

39. ANIMAL CELL CULTURE SYSTEM
 It is expression of recombinant proteins in animal cell.
 Therapeutic proteins are produced from animal cells by genetic
manipulation through advances techniques.
 Mammalian cell culture is technically more complex and expensive
than microbial cell fermentation
 Therefore, it is usually only used in the manufacture of therapeutic
proteins that are extensive and show essential post-translational
modifications
 Advantage of animal cell culture in therapeutic is ability to carry out
post translational modification of protein product.
 Due to this many biopharmaceuticals is produced in animal cell lines
Example. -CHO(Chinese hamster ovary) and
-BHK(baby hamster kidney) cells are the most popular
-Hybridoma cells in the case of some monoclonal antibodies

40. Steps in animal cell culture

41.  In addition to recombinant biopharmaceuticals, animal cell culture
is used to produce various other biologically based
biopharmaceuticals such as:
 vaccines and hybridoma cell-produced monoclonal antibodies.
 Lymphoblastoid cell line (the Namalwa cell line) produced
interferons, to synthesize high levels of several IFN-α’s naturally
Disadvantages of Animal cell-based culture systems
 Animal cell-based systems has many disadvantages when compared
with E. coli
 This include:-
 Animal cells display a very complex nutritional requirement
 They grow more slowly and
 more susceptible to physical damage b/c they don’t have cell
wall,
 This increased production cost

42. Applications of cell line/ animal cell culture
Recombinant Vaccines Production
Virus cultivation and study
Cellular and molecular biology
In Cancer Research
Gene therapy
Immunological studies
Others: Cell lines are also used in
 in-vitro fertilization (IVF) technology,
recombinant protein and drug selection and
improvement.
 Production of MAb - Hybridoma
 Tissue engineering - artificial organs
 In vitro cell toxicity and drug screening

43. Comparison of animal cell culture and microbial cell
culture.
The culture of animal cells differs from that of
microbial cells in:-
They require more complex media
Extended duration of fermentation due to slow growth
of animal cells
They are more fragile than microbial cells due to the
absence of an outer cell wall.
Antibiotics Supplemental serum is required.
Reactor design and operation differs somewhat from
microbial fermentations.

45. Applications of transgenic plat
and animals in biopharmaceutical
production is:-
#ASSIGNMENT

46. END OF
UNIT 1&2
THANK YOU
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