Presented by - R Hemamalini 2010CYZ8795 Group G1

2.

Market Scenario

Sources

Technologies for production

Enzyme Replacement Therapy

Enzyme treatment for special conditions

Introduction

Enzymes are biological catalysts, usually proteins that

accelerate chemical reaction rates by a million fold or more.

The hallmarks of enzyme action are specificity, and

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.

3.

High affinity

Specificity

Potent

Incentives

4.

Need encapsulation

Long-term dependency

High Purity

Immunogenic

5.

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.

6.

Enzymes in

medicine

EnzymeReplacement

TherapyProphylaxis

Treatingspecial

conditions

Extraction of

medically important compounds

DiagnosisDrug

manufacture

CancerWound healing

Lactose intolerance

Clotting Anti-clotting

7.

Deficient enzyme activity

Accumulation of key products or substrates ERT

Clinical symptoms

Alleviation of symptoms

8.

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

9.

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

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.

11.

Adenosine and deoxyadenosine accumulate

inhibit S-adenosyl homocysteine hydrolase

accumulation of SAH

failure of methylation reactions

(important for detoxification of adenosine and deoxyadenosine)

12.

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

Sucrase – isomaltase EC 3.2.1.10(/48) 13.

14.

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

Phenylalanine monooxygenase (EC 1.14.16.1)

15.

16.

Gaucher’s

disease

Found by

Phillipe Gaucher

in 1882

Biochemical

basis for the

disease in 1965

by Brady et al..

Glucosyl

CH2-CH-CH-CH=CH-(CH2)12-CH3

O=C-CH2-CH2-CH2-(CH2)n-CH3

N OH

Ceramide

OH-CH2-CH-CH-CH=CH-(CH2)12-CH3

O=C-CH2-CH2-CH2-(CH2)n-CH3

N OH

Glucose Ceramide

Glucocerebrosidase - EC 3.2.1.45 Glucocerebrosidase - EC 3.2.1.45

Cerebrosidase /β-glucosidase

17.

The lack of the glucocerebrosidase, a β- glucoronidase

leads to accumulation of glucocerebroside.

Autosomal recessive disorder

Ceredase - ERT with β- 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.

DIGESTIVE SYSTEMIC

Amylase

Lipase

Protease

Lactase

Cellulase

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

UrokinaseTrypsin

Chymotrypsin

22.

Microbial production

Mammalian cell processes

Plant cultivation

Recombinant DNA technology

23.

Source:- Protein expression workflow by Sigma-Aldrich

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

24.

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 policies to promote academic-industry

interaction.

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

25.

1. BCC Research on Medical Enzymes: Technology and Global Markets

2. 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. 2339–2344, 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. Jennifer Hammer, Stan Bynum, Therapeutic Use of Enzymes for Vollara

8. Michael Vellard, The enzyme as drug: application of enzymes as pharmaceuticals, Current

Opinion in Biotechnology 2003, 14:1–7

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 Noël 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. Protein expression workflow by Sigma-Aldrich

11. 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. The Enzyme Revolution by Enzymedica

15. 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