Presented by -

R Hemamalini

2010CYZ8795

Group G1

2.

Introduction Enzyme Replacement Therapy

Sources Enzyme treatment for special conditions Market Scenario Technologies for production

3.

Enzymes The

are

biological of

catalysts, action

usually are

proteins

that and

accelerate chemical reaction rates by a million fold or more. hallmarks enzyme specificity, efficiency. They are essential for cell and tissue viability. Clinical consequences result in their absence or malfunction.

Therapeutic enzymes are those enzymes which can be safely

used in medicine to cure diseases and medical conditions effectively and safely. Medication may involve only the

concerned enzyme or in combination with other therapies.

4.

Specificity

Incentives

High affinity

Potent

5.

Long-term dependency

Immunogenic

Need encapsulation

High Purity

6.

The use of animal enzymes instead of microbial enzymes, although these would be costlier. Modified by covalent attachment of polyethylene glycol, for e.g in asparaginase Entrapment of the enzyme within artificial liposomes, synthetic microspheres and red blood cell ghosts. Choose the sources carefully to avoid any unwanted

contamination by incompatible material and to enable ready

purification. Use prophylactic enzymes as a short-term aid only.

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Enzymes in medicine

Enzyme Replacement Therapy

Prophylaxis

Treating special conditions

Extraction of medically important compounds

Diagnosis

Drug manufacture

Cancer

Wound healing

Lactose intolerance

Clotting Anti-clotting

8.

Deficient enzyme activity

Accumulation of key products or substrates ERT Clinical symptoms

Alleviation of symptoms

9.

Adequate drugs for many of rare diseases and conditions have not been developed Orphan drugs Passed in 1983 in the United States Encourage pharmaceutical companies to develop treatments for

diseases affecting only small numbers of people (less than 200

000). In Europe and Australia, there is comparable legislation that

provides similar protection and incentives

Source:- Stefano Villa, Amelia Compagni and Michael R. Reich (2008) Orphan drug legislation: lessons for neglected tropical diseases, Int J Health Plann Mgmt,2008

10.

Adenosine deaminase - EC 3.5.4.4

11.

Severe Combined Immunodeficiency (SCID) Autosomal recessive, frequent infections result in early death.

Intracellular

purine

metabolites

accumulate

to

toxic

concentrations within cell. T-cell and B-cell function absent from birth ADAGEN (Pegademase Bovine) Enzon Inc. Sigma-Tau Pharmaceutical Inc. modified enzyme for ERT derived from bovine intestine injections rh ADA developed by R&D systems, Minneapolis, but for research only, Feb, 2011.

12.

Adenosine and deoxyadenosine accumulate

inhibit S-adenosyl homocysteine hydrolase

accumulation of SAH

failure of methylation reactions (important for detoxification of adenosine and deoxyadenosine)

Sucrase isomaltase EC 3.2.1.10(/48)

13.

Congenital sucrase-isomaltase deficiency (CSID) An autosomal recessive disease of the small intestine first discovered in 1960 by Weijers and colleagues. Sucrase and maltase activity absent or minimal. Sucrase replacement therapy with 100-fold more potent enzyme

Sucrosidase - Sucraid
Liquid form, Saccharomyces cerevisiae-derived, oral solution 8,500 I.U per ml of sucrase

Orphan drug in 1998 by FDA

14.

Phenylalanine monooxygenase (EC 1.14.16.1)

15.

Deficiency of the above enzyme -> Phenylketonuria (PKU) Genetic disease Treatment from birth with a low phenylalanine diet. Phenyl alanine ammonia lyase - EC 4.3.1.5 acts as a substitute for the deficient enzyme. PAL,converts phenylalanine to trans-cinnamic acid, a harmless metabolite. Sources: Plants, recombinant Modified with PEG for stability

16.

Glucocerebrosidase - EC 3.2.1.45
17.

Gauchers disease Found by Phillipe Gaucher in 1882

Glucosyl
O=C-CH2-CH2-CH2-(CH2)nCH3 OH N CH2-CH-CH-CH=CH-(CH2)12-CH3

Ceramide Cerebrosidase / -glucosidase

Biochemical basis for the

disease in 1965
by Brady et al..
O=C-CH2-CH2-CH2-(CH2)nCH3 OH N Glucose OH-CH2-CH-CH-CH=CH-(CH2)12CH3 Ceramide

The lack of the glucocerebrosidase, a - glucoronidase leads to accumulation of glucocerebroside. Autosomal recessive disorder Ceredase - ERT with - glucoronidase extracted from

human placental tissues, from Genzyme

Cerezyme recombinant human enzyme expressed in chinese hamster ovary cells from Genzyme. Annual cost of ERT: $80,000 for each patient annually. Administered as injections.

19.

20.

Cellulase

Amylase Lipase Protease Lactase DIGESTIVE SYSTEMIC

Nattokinase Serratio-

peptidase
Proteolytic

Source:- The Enzyme Revolution by Enzymedica

21.

Bilirubin oxidase

Heparinase
Sucrase-isomaltase Ribonuclease T1 L-Asparaginase Collagenase Streptokinase Serratiopeptidase Uricase

Papain Bromelain Phenylalanine ammonia lyase Nattokinase Amylase

Glucocerebrosidase DNase Urokinase Alpha Glucosidase Hyaluronidase Urokinase Trypsin Chymotrypsin

22.

Microbial production

Plant cultivation

Mammalian cell processes

Recombinant DNA technology

23.

Source:- Protein expression workflow by Sigma-Aldrich

24.

The global market for medical enzymes was estimated at $6

billion in 2010.
The market is growing at a compound annual growth rate (CAGR) of 3.9%, to reach $7.2 billion in 2015.

Therapeutic enzymes are the biggest segment in terms of

revenues generated- valued at $5.3 billion in 2010 and expected to reach $6.3 billion in 2015, at a 3.6% compound annual growth

rate (CAGR) .
Enzymes used in molecular research will experience the fastest compound annual growth rate (CAGR), 6.2%, over the study

period. This sector was worth $546 million in 2010 and should
reach nearly $739 million by 2015.
Source: BCC Research on Medical Enzymes: Technology and Global Markets

25.

Total

enzyme

consumption

figures

of

India

are

comparatively low: A major difference is noticed in the

enzyme consumption pattern, compared to world figures,

e.g. the use of enzyme in detergents is much smaller. Special efforts are needed to formulate favourable

government
interaction.

policies

to

promote

academic-industry

Some companies like: Advanced Enzymes, Biocon, Bharat

Pharma are producing therapeutic enzymes.

Source: TIFAC Report on Bioenzymes: Production technology Website of Advanced Enzymes, Biocon and Bharat Pharma

1. 2.

BCC Research on Medical Enzymes: Technology and Global Markets Cerezyme Imiglucerase for Injection http://www.cerezyme.com/~/media/Files/CerezymeUS/pdf/cerezyme_pi.pdf

3.

Christineh N. Sarkissian, Zhongqi Shao, Franc Oise Blain, Rosalie Peevers, Bongsheng Su,
Robert Heft, Thomas M. S. Chang and Charles R. Scriver, A different approach to treatment of phenylketonuria: Phenylalanine degradation with recombinant phenylalanine ammonia lyase, Proc. Natl. Acad. Sci. USA Vol. 96, pp. 23392344, March 1999

4.

D. R. HeadonAnd G. Walsh, The industrial production of enzymes, Biotech. Adv. Vol. 12, pp.

635--646,1994
5. David J. Saul, Moreland D. Gibbs and Peter L.Bergquist, Biocatalysis: Industrial Enzymes and the exploitation of micro-organisms, New Zealand Institute of Chemistry 6. Enzymes: EC Nomenclature 4th level Complete List of all Enzymes, http://www.biologie.uni-hamburg.de/b-online/e18_1/ec4.htm

7.
8.

Jennifer Hammer, Stan Bynum, Therapeutic Use of Enzymes for Vollara

Michael Vellard, The enzyme as drug: application of enzymes as pharmaceuticals, Current Opinion in Biotechnology 2003, 14:17

9.

Michel Duval, Stefan Suciu, Alina Ferster, Xavier Rialland, Brigitte Nelken, Patrick Lutz, Yves Benoit, Alain Robert, Anne-Marie Manel, Etienne Vilmer, Jacques Otten and Nol Philippe, Comparison of Escherichia coli-asparaginase withErwinia-asparaginase in the treatment of childhood lymphoid malignancies: results of a randomized European Organisation for Research and Treatment of Cancer--Children's Leukemia Group phase 3 trial, Blood, 15 April 2002 Volume 99, Number 8

10. 11.

Protein expression workflow by Sigma-Aldrich Rodney J.Y Ho, Milo Gibaldi (2003) in Biotechnology and Biopharmaceuticals, Wiley-Liss, NJ, pp. 245-269

12.

Stefano Villa, Amelia Compagni and Michael R. Reich (2008) Orphan drug legislation: lessons for neglected tropical diseases, Int J Health Plann Mgmt,2008

13.

Syndey Sandberg, Special Report: A nutraceutical approach to enhancing memory, Enzyme News

14. 15.

The Enzyme Revolution by Enzymedica Walsh, G (2003) in Biopharmaceuticals Biochemistry and Biotechnology, Wiley, West Sussex, 2nd Ed., pp. 351-402

Dr. Ravi Krishnan Elangovan for inspiring me to take up this topic Prof. S.K. Khare for his support and guidance

My colleagues and friends for their help and critically

reviewing the content My batchmates for helping me use their resources Family for their patience and understanding