27

PROTEIN THERAPEUTICS

Embed Size (px)

Citation preview

EVALUATION SEMINAR ON PROTEIN THERAPEUTICS

PRESENTED BY:

Ms. PRIYANKA SHRESTHA

1ST M. PHARM

DEPT. OF PHARMACOLOGY

FACILIATED TO:

DR. SHIVALINGE GOWDA K.P.

H.O.D, DEPARTMENT OF

PHARMACOLOGY

TABLE OF CONTENTS  DEFINITION

ATTRACTIVE FEATURES OF PROTEIN MOEITY

PROTEIN DISCOVERY: HISTORY AND EVOLUTION

SCOPE OF PROTEIN THERAPEUTICS

CLASSIFICATION OF PROTEIN THERAPEUTICS

PRODUCTION OF THERAPEUTIC PROTEINS

OBSTACLES IN PRODUCTION

SOURCES

EXAMPLES OF PRODUCTION

DELIVERY SYSTEM

EXAMPLES OF SOME MARKETED PROTEIN THERAPEUTICS

APPLICATION AND MARKET OF PROTEIN THERAPEUTICS

REFERENCE

THERAPEUTIC PROTEIN

Proteins which are engineered in the laboratory for pharmaceutical use are referred to as therapeutic proteins.

Proteins which are absent or low in individuals with an illness such as Cancer, Infectious diseases, Hemophilia, Anemia, Multiple sclerosis, Hepatitis B/C, etc. are artificially synthesized on large scale through genetically modified host cells and delivered.

PROTEIN THERAPEUTICS

This therapeutic approach in treating diseases using proteins

and peptides is termed protein therapeutics.

Protein therapy is similar to gene therapy, but unlike gene

therapy, protein therapy delivers protein to the body in

specific amounts (as would be ordinarily present), to help

repair illness, treat pain or remake structures.

Introduced in 1920’s Human insulin is considered to be the

first therapeutic protein.

ATTRACTIVE FEATURES Proteins have been considered for the following facts

Diversity of functional groups: free thiols (on cysteine

residue) & amine (on the N-terminus or on lysine residue)

Imitations by simple chemical compounds are less

Lower side effects: due to high specificity there’s less

potential for protein to interrupt the normal biological

processes

Less likely for the body to evoke immune responses as the

body naturally produces many of the proteins

Clinical development and FDA approval time are

comparatively faster than that for small molecule drugs.

PROTEIN DISCOVERY: HISTORY

Proteins were accepted as a distinct class of biological

molecules in the 18th century by Antoine Fourcroy and other.

They are found to be able to coagulate in distinct conditions

e.g. albumen from egg whites, blood serum albumin, fibrin,

and wheat gluten.

The elemental analysis of protein by Gerhardus Johannes

Mulder & use the name ‘Protein’ coined by JönsJakob

Berzelius in ~1839 in his papers.

The evolution of Protein therapeutics

1953: First accurate model of DNA was suggested

1982: Recombinant DNA technology was used to create human insulin

1986: Approval of Interferon Alfa and muromonab-CD3

1993: CBER's Office of Therapeutics Research and Review (OTRR) was

formed

1997: First whole chimeric antibody, Rituximab, and first humanized

Antibody, Daclizumab, approved

2002: Market for biotechnology products represented approximately

$30 Billions to $400 billion in yearly worldwide pharmaceutical sales

2006: An inhaled form of insulin (Exubera) was approved, expanding

protein Products into a new dosage form

SCOPE OF PROTEIN THERAPEUTICS

The hope is that the protein, which is not present in adequate

levels, will function as it is designed to do.

For example, use of certain proteins in addressing

cardiovascular disease has been evaluated in some studies.

Especially when veins or arteries become blocked, the right

types of proteins might help here by building new passages for

blood flow.

Some doctors suggest that protein therapy of this type might

eventually be so successful that it could eliminate the need for

complicated surgeries like bypass surgery.

CLASSIFICATION

Classification based on pharmacological action:

Group I: protein therapeutics with enzymatic or regulatory

activity

I a: Replacement of a protein that is deficient or abnormal: e.g. -

Exubera, Increlex

I b: Augmentation of an existing pathway: e.g. - Ovidrel , Neupogen

I c: Provides a novel function or activity: e.g. - Myoblock

Group II : protein therapeutics with special targeting activity

II a: Interferes with a molecule or organism: e.g. - Avastin

II b: Delivers other compounds or proteins (such as radionuclie,

cytotoxic drug or effector protein): e.g. – Ontak

Group III : Protein vaccines

IIIa : Protecting against a deleterious foreign agent: e.g. - Engerix

IIIb : Treating an autoimmune disease. : e.g. - Rophylac

Group IV : Protein diagnostics: e.g. – Geref

Classification based on molecular types:

Antibody based drugs, Fc fusion proteins, anticoagulants, blood

factors, growth factors, hormones, interferon, bone morphogenetic

proteins, interleukins and thrombolytic.

Classification based on molecular mechanism:

Binding non-covalently to target e.g. –mAbs

Affecting covalent bonds e.g. – enzymes

Exerting activity without specific interactions e.g. - serum albumin

PRODUCTION OF THERAPEUTIC PROTEINS

OBSTACLES IN PRODUCTION:

Protein solubility, distribution, stability

Delivery route (blood or digestive juice may degrade it)

Physiologically active-post translational modifications

Cost

SOURCES

Various types of cells like

bacterial cells,

yeast cells,

insect cells,

microbes (Bacillus sp., Actinomycetes sp., E coli),

plants,

animals,

also mammalian cells

are used for producing therapeutic proteins under defined conditions.

EXAMPLE OF PRODUCTION

MANUFACTURING SYNTHETIC HUMAN INSULIN

51 amino acids = two chains

linked by disulfide bonds

21 in chain A

30 in chain B

DIAGRAMMATIC REPRESENTATION

Synthesis of the DNA containing the nucleotide sequences of

the A and B polypeptide chains of insulin. 

Plasmid + restriction enzyme Insertion of the insulin gene into

plasmid (circular DNA)  

Restriction enzymes cut plasmidic DNA

DNA ligase agglutinates the insulin gene and the plasmidic

DNA Plasmid + insulin gene

Introduction of recombinant plasmids into bacteria: E. coli

E.coli = factory for insulin production

Using E. coli mutants to avoid insulin degradation

Bacterium reproduces the insulin gene replicates along

with plasmid E. Coli

Formed protein partly of a byproduct the A or B chain of

insulin

Extraction and purification of A and B chain

Connections of A- and B-chain by reaction forming disulfide

cross bridges

results in Pure synthetic human insulin

Example of production explained

Production of recombinant protein therapeutics in cultivated mammalian cells

Production of recombinant protein therapeutics in milk of transgenic animals and schematic representation of the process used to purify atryn from the milk of transgenic goats

DELIVERY SYSTEM

Proteins with a peptide sequence that shows the capability

to translocate membrane rapidly, termed as ‘‘cell

penetrating peptide (CPP) or protein transduction

domain(PTD) are covalently modified by methods

mentioned below:

1. Recombinant fusion protein from a vector containing DNA

sequence of the CPP sequence are directly expressed

2. Linker such as disulfide bond linkage that is cleavable

under reductive environment are used for protein or

chemical conjugation of CPP to the protein

Peptide based biomaterial are used for delivery to protect

protein from protease degradation & Strategy to improve

delivery efficiency for the following reasons :

-Easy to synthesize -Easy characterization -Less toxic & has

higher immunogenicity than high molecular wt polymers.

Due to its amphipathic character of peptides can associate

rapidly with protein cargos in solution in self-assembly manner,

possibly through noncovalent hydrophobic interaction.

Protein polymers conjugate for Targeted delivery

Most commonly employed polymer : Polyethylene

glycol(PEG) & Poly(N-isopropyl acrylamide) (PNIPAM)

These polymers that alter their solubility or propensity for

self-assembly when exposed to changes in pH or

temperature allow their responsive nature to be conferred

to the protein to which they are attached.

Functionalizable with active esters & hence can be

conjugated with protein amine.

 

MARKETED PRODUCTS

rhDNase IFOLLICLE STIMULATING HORMONE (FSH)

APPLICATIONS

The therapeutic proteins in market and clinical trials today are

produced by recombinant method.

Higher efficiency and fewer side effects of protein therapeutics are

driving the protein therapeutics market comprising 16% of prescription

drug sales in 2011 of which antibody-based drug accounts to be the

fastest growing class of protein therapeutics.

REFERENCES: Protein therapeutics: a summary and pharmacological classification Benjamin Leader,

Quentin J. Baca and David E. Gola

Production of recombinant protein therapeutics in cultivated mammalian cells Florian M

Wurm

Recombinant Proteins Amith Reddy Eastern New Mexico University

Recombinant therapeutic proteins Beenish Choudhary

DESIGN AND PURIFICATION OF PROTEINS Biotechnology project, 18/05/09 Marielle

Brockhoff, Aurore Lacas , Raphael Lieberherr Sebastian Olényi, Morgane Perdomini,

Zrinka Raguz.

production of recomb. Proteins BRUNO THADEUS

Protein therapeutics Soma Mukherjee SMU, Chemistry 5th April’2011

http://en.wikipedia.org/wiki/Protein_therapy

Protein_Therapeutics--Xiao_Yu.ppt

THANK YOU..!