PRODUCTION OF PHARMACEUTICALS BY GENETICALLY ENGINEERED CELLS (HORMONES AND INTERFERONS)
1. PRODUCTION OF PHARMACEUTICALS BY GENETICALLY
ENGINEERED CELLS (HORMONES AND INTERFERONS)
Prabhu Thirusangu
Molecular Biomedicine Laboratory,
Sahyadri Science College,
Kuvempu University
2. THE AGE OF BIOPHARMACEUTICALS….
The term ‘biopharmaceutical’ was first used in the 1980s and came to
describe a class of therapeutic protein produced by modern
biotechnological techniques, specifically via genetic engineering.
Biomedical research continues to broaden our understanding of the
molecular mechanisms underlining both health and disease.
Research undertaken since the 1950s has pinpointed a host of proteins
produced naturally in the body which have obvious therapeutic
applications.
Examples include the interferons, and interleukins, which regulate the
immune response; growth factors such as erythropoietin, which
stimulates red blood cell production; and neurotrophic factors, which
regulate the development and maintenance of neural tissue.
3. Production of Human Insulin by r- DNA Technology….
Human insulin produced by recombinant DNA technology was first
approved for general medical use in 1982.
It was the first product of recombinant DNA technology to be approved
for therapeutic use in humans.
The quantity of purified insulin obtained from the pancreas of one pig
satisfies the requirements of one diabetic for 3 days.
6. Route to the
Production by
Bacteria of
Human Insulin
Overviewof gene
cloning.
Cloning
animation
7. 3D- Structure of engineered Insulin
Recombinant Insulin
(Humulin)
8. ENGINEERED HUMAN INSULIN…
Product Company
Humulin (rh-insulin)
Novolin (rh-insulin)
Humalog (Insulin Lispro, an insulin analogue)
Insuman (rh-insulin)
Liprolog (Bio Lysprol, a short-acting insulin)
NovoRapid (Insulin Aspart, short-acting rh-insulin analogue)
Novomix 30 (contains Insulin Aspart, short-acting rh-insulin
analogue (see NovoRapid) as one ingredient
Novolog (Insulin Aspart, short acting rh-insulin; see also
NovoRapid entry above)
Novolog mix 70/30 (contains Insulin Aspart, short acting rh-
insulin analogue as one ingredient; see also Novomix 30 entry
above)
Actrapid/Velosulin/Monotard/Insulatard/Protaphane/Mixtar
d/Actraphane/ Ultratard (all contain rh-insulin produced in
S. cerevisiae, formulated as short/intermediate/long-acting
products)
Lantus (insulin glargine, long acting rh-insulin)
Optisulin (insulin glargine, long-acting rh-insulin analogue,
see Lantus)
Eli Lilly
Novo Nordisk
Eli Lilly
Hoechst AG
Eli Lilly
Novo Nordisk
Novo Nordisk
Novo Nordisk
Novo Nordisk
Novo Nordisk
Aventis Pharmaceuticals
Aventis Pharma
9. PRODUCTION OF recombinant HUMAN GROWTH HORMONE
(Hgh)….
Human growth hormone is a polypeptide hormone synthesized in the anterior
pituitary. It promotes normal body growth and lactation and influences various
aspects of cellular metabolism.
Dwarfism caused by insufficient production of HGH by the pituitary gland.
HGH can treat dwarfism – to help under sized children reach their normal
height and size.
Recombination process –
1. DNA coding for growth hormone is inserted
into the plasmid.
2. Introduction of recombined plasmid into E.Coli host
cell
3. Engineered E.Coli cell is allowed to multiply in the fermentor.
11. Cytokines constitute the single most important group of biopharmaceutical
substances. As coordinators of the immune and inflammatory response,
manipulation of cytokine activity can have a major influence on the body’s
response to a variety of medical conditions.
Administration of certain cytokines can enhance the immune response against a
wide range of infectious agents and cancer cells.
12. THE INTERFERONS…..
Interferons (IFNs) were the first family of cytokines to be discovered. In
1957 researchers observed that if susceptible animal cells were exposed to
a colonizing virus, these cells immediately become resistant to attack by
other viruses.
This resistance was induced by a substance secreted by virally-infected
cells, which was named ‘interferon’ (IFN).
Humans produce at least three distinct classes, IFN-a, IFN-b and IFN-g.
Biological effects,
Induction of cellular resistance to viral attack.
Regulation of most aspects of immune function.
Regulation of growth and differentiation of many cell types.
Sustenance of early phases of pregnancy in some animal species.
13. PRODUCTION OF INTERFERONS BY GENETIC
ENGINEERING…
A DNA sequence coding for the product was synthesized and inserted
into E. coli. The recombinant product accumulates intracellularly as
inclusion bodies
Large-scale manufacture entails an initial fermentation step. After
harvest, the E. coli cells are homogenized and the inclusion bodies
recovered via centrifugation. After solubilization and refolding, the
interferon is purified to homogeneity by a combination of
chromatographic steps.
The final product is formulated in the presence of a phosphate buffer
and sodium chloride.
It is resented as a 30 mg/ml solution in glass vials and displays a shelf-
life of 24 months when stored at 2–8°C`.
16. 3D – Structure of Human growth hormone
Recombinant interferons
17. Interferon toxicity…..
Like most drugs, administration of IFNs can elicit a number of
unwanted side effects.
Minor side effects - Range of flu-like symptoms, e.g. fever, headache,
chills.
Serious potential side effects –
Anorexia
Strong fatigue
Insomnia
Cardiovascular complication
Autoimmune reactions
Hepatic decompression
18. CONCLUSION…..
Interferons represent an important family of biopharmaceutical
products.
They have a proven track record in the treatment of selected medical
conditions, and their range of clinical applications continue to grow.
It is also likely that many may be used to greater efficacy in the future
by their application in combination with additional cytokines
Prabhu Thirusangu
19. Reference…
Bibliography……
Biopharmceuticals (2nd Edition) By Gary Walsh
Immunology By Kuby
Genetics By Benzamin
Webliography……
http://www.ncbi.nlm.nih.gov/pubmed/3296632
http://en. Wikipedia.com/pharmacology
