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    • Chapter 14:Application of Recent Advances inImmunology for Developing NovelBiotherapeutics in IndiaKumar Shah, MD
    • Advances inBiopharmaceutical Technology in India January 2008 Editor: Eric S. Langer BioPlan Associates, Inc. Rockville, MD, USA SOCIETY FOR INDUSTRIAL MICROBIOLOGY Arlington, VA, USA
    • BioPlan Associates, Inc.15200 Shady Grove Road, Suite 202Rockville MD 20850 USA301-921-9074www.bioplanassociates.comandSociety for Industrial Microbiology3929 Old Lee Highway Suite 92AFairfax, VA 22030-2421703-691-3357Copyright © 2008 by Eric S. LangerAll rights reserved, including the right of reproduction in whole or in part in any form. Nopart of this publication may be reproduced, stored in a retrieval system, or transmitted inany form or by any means, electronic, mechanical, photocopying, recording or otherwise,without the written permission of the publisher.For information on special discounts or permissions contact BioPlan Associates, Inc. at301-921-9074, or info@bioplanassociates.comEditor: Eric S. LangerProject Director, Enterprise: Yibing (Eliza) ZhouProject Director, Scientific: John MorrowProduction: ES Illustration and Design, Inc.Text and Cover Design: Esperance ShatarahCover photos courtesy of Shantha BiotechISBN 978-1-934106-07-5 ii
    • AcknowledgmentThis project would not have been possible without the exceptional efforts of the many peopleinvolved. In particular, we would like to thank: Yibing (Eliza) Zhou, Project Director, Enterprise John Morrow, Project Director, Scientific ES Illustration and Design, Inc., Production Manager, cover graphics.We would also especially like to thank our reviewers, whose expertise ensured this volumeaddressed today’s most important issues: Prof. P.S. Biesen, Director, Madhav Institute of Technology and Science, India Prof. Ananda Chakrabarty, University of Illinois, USA Dr. Thomas E. Colonna, President, Biotech Consultant LLC, USA Dr. John Curling, President, John Curling Consulting, Sweden Dr. Milind Deshpande, Technical Director, Fermentation and Processing, University of Iowa, USA Dr. P.K. Ghosh, President, Biotechnology, Cadila Pharmaceuticals Ltd., India Dr. Vijai Kumar, President & Chief Medical Officer, Excel Life Sciences, Inc., USA Dr. Yennapu Madhavi, Scientist, National Institute of Science Technology and Development Studies, India Dr. John Morrow, President, Newport Biotech, USA Dr. Brandon Price, President. Falconridge Associates, Inc., USA Dr. Gayatri Saberwal, Scientist, Institute of Bioinformatics and Applied Biotechnology (IBAB), India Dr. V.K. Srinivas, General Manager R&D, Bharat Biotech, India Dr. S. Vsisalakshi, Scientist, National Institute of Science Technology and development Studies, India Dr. Scott Wheelwright, President, Strategic Manufacturing Worldwide, Inc., USAWe also extend our sincere appreciation to Shantha Biotech, and Ms. Sravanthi Reddy for hersupport and use of cover photos and graphics.Thank you for your efforts, and your recognition of the importance of this study.Eric S. LangerManaging Editor iii
    • PrefaceT his study was undertaken, managed and coordinated by BioPlan Associates, Inc., a biopharmaceutical management and marketing research consulting firm in Rockville, MD, based on nearly 20 yearsexperience and knowledge of the market segment. BioPlan surveyed theindustry to identify required content, and then selected subject matter expertsto author relevant chapters to this study.The Society for Industrial Microbiology (SIM), in recognizing the importanceof applied sciences in biotechnology processes, has lent its name to thisendeavor. The Society for Industrial Microbiology is a nonprofit professionalassociation dedicated to the advancement of microbiological sciences,especially as they apply to industrial products, biotechnology, materials,and processes. Founded in 1949, SIM promotes the exchange of scientificinformation through its meetings and publications, and serves as liaison amongthe specialized fields of microbiology. Membership in the Society is extendedto all scientists in the general field of microbiology.India is one of the fastest growing economies in the world. The country hasinvested heavily in advancing its pharmaceutical and biopharmaceuticaltechnologies to improve its healthcare systems, its population’s general health,and its overall economy.Both scientists and entrepreneurs in India have made important contributionsto advancing the field at many levels. This study provides a framework fromwhich both those new to India’s rapid advancements in biotherapeutics andvaccines, and those with long histories can recognize the potential, and planfor the future. The findings of this study support worldwide public health andeconomic policy.Each chapter provides unbiased, peer-reviewed perspectives of the currentstate of the science and technology associated with biopharmaceuticals inIndia. While no single work can encompass all the advances being made inthe field, this study offers a comprehensive assessment of the technological andeconomic advancements in India.The intended audiences include decision-makers at biopharmaceuticalresearch organizations, biotherapeutic manufacturers, contract manufacturingorganizations, suppliers to the industry, policy-makers, and internationalentities evaluating this market. We plan to keep this study current by providingregular updates as technologies, and the industry advance.iv
    • Advances in BiopharmaceuticalTechnology in India Table of Contents PART 1: Introduction .............................................................viiChapter 1 Prospects for Modern Biotechnology in India ....................................................... 1 Prasanta K. Ghosh, Prasenjeet Ghosh, Soma Ghosh, and Kushal Shodhan PART 2: State of India’s Life Sciences Industry ......................67Chapter 2 A Window into India’s Biopharma Sector .............................................................69 Narayan KulkarniChapter 3 Indian Biopharmaceutical System and Policies ................................................109 Yennapu Madhavi, Ph.D.Chapter 4 Government Support for Biotech and Biopharmaceuticals Industry ..........127 Dr. B.M. GandhiChapter 5 Biopharmaceutical Products in India ..................................................................179 Scott M. Wheelwright, Ph.D., and Hazel Aranha, Ph.D.Chapter 6 Biogeneric Manufacturing in India ......................................................................203 Ashesh Kumar, Ph.D.Chapter 7 Biopharmaceutical Market Situation ..................................................................235 Merlin H. Goldman, PhD MBA CEng MIChemEChapter 8 The Contract Research Industry in India ............................................................281 Umakanta Sahoo, MBA, Ph.D., and Faiz Kermani, Ph.D.Chapter 9 Regulatory Landscape for Clinical Trials in India ..............................................313 Romi Singh, Ph.D. and Brijesh RegalINDIA BRIEF 1 Pre-Clinical and Clinical Trial in India ............................................................................................. 327 V.K.Srinivas, Ph.DChapter 10 Life Sciences Education in India...................................................................................333 Dr Dipti Sawant, Ph.D., MBA, CCRA v
    • PART 3: Scientific Issues in India .........................................383Chapter 11 Vaccine Industry in India .......................................................................................385 Dr. Prasad S. Kulkarni, M.D. and Suresh S. Jadhav, Ph.D.Chapter 12 Development of Plasma-Derived Biopharmaceuticals in India: Challenges and Opportunities .............................................................................465 John M. Curling, Christopher P. Bryant, Timothy K. Hayes, Ranjeet S. AjmaniChapter 13 Bioprocess Expression and Production Technologies in India .......................519 David M MousdaleINDIA BRIEF 2 Technology Transfer: Impact and Importance for Indian Biotechnology Growth ........ 555 Susan Kling FinstonChapter 14 Application of Recent Advances in Immunology for Developing Novel Biotherapeutics in India.........................................................................................563 Kumar Shah, MDChapter 15 Early-stage Key R&D Components for Successful Development of Protein Therapeutics ..............................................................................................589 Krishna M. Madduri, Ph.D.Chapter 16 Understanding Ayurveda Medicine ....................................................................605 Dr. Sampada Amol Mahajan PART 4: Biobusiness in India ...............................................619Chapter 17 Biopharmaceutical Outsourcing: A Comparative Overview of the Landscape between India and China ...........................................................621 Minna A. DamaniChapter 18 Biopharmaceutical Research Collaborations between India and the West: A Guide to Prospective Partnerships .................................................661 Uri Reichman, Ph.D., M.B.A., Bharat Khurana, D.V.M., Ph.D., and Steven M. FergusonChapter 19 Outsourcing R&D to India......................................................................................723 Probir Roy Chowdhury and Sajai SinghINDIA BRIEF 3 Outsourcing Biopharma R&D to India........................................................................................... 741 Jim Schnabel, Senior Research Associate, BioPlan Associates, Inc.Chapter 20 Venture Capital in India Today..............................................................................747 Sajai Singh and Probir Roy Chowdhury vi
    • 14 Application of Recent Advances in Immunology for Developing Novel Biotherapeutics in India KUMAR SHAH, MD, President Amit Shah, Scientific Officer Department of Research and Development Endocrine Technology, LLC 304 Livingston Street, Brooklyn, NY 11217 Phone: 718-222-1065 Fax: 718-852-7412 E-mail:
    • Advances in Biopharmaceutical Technology in India About the Authors KUMAR SHAH, MD is a Founder and President of Endocrine Technology, LLC in NY, USA. He has over 20 publications and several patents related to the modulation of Factor H. He is a Diplomat in Internal medicine, Diabetes, Endo- crinology and Metabolism. He is in active consulting practice. In India he served as clinical research physician for Roche Products and acted as a consultant to initiate “Cell Transplant Laboratory for the cure of diabetes” for the institute of kidney diseases, Ahmedabad, Gujarat State. He has presented paper in National HIV/AIDS conferences held in Mumbai (HIV Congress: 11-13th March, 2005). AMit SHAH is a scientific officer of Endocrine Technology, LLC. He is involved in developing immune modulation technology of the Company and in the management of information technology. He has a degree in computer sciences and research interests in biotechnology field. 564
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel Biotherapeutics ABSTRACT I mmunity is a major barrier in the translational development of bio-pharmaceutical drugs. The US Food and Drug Administration estimates a cost saving of US$100 Million if the human safety of new drugs could be predicted with 10% accuracy. The recent advances in the fundamentals of immunology have assigned a host protective role to Factor H. Factor H is an immune regulatory protein of a billion year old early human defense system such as Alternate Complement System. The Alternate Complement System plays a crucial role in activating host inflammatory and memory responses when foreign material such as bacteria, viruses, cancer cells, allogenic and xenogenic somatic cells, biotherapeutics drugs or bioterrorism related organisms interact with host and its immune system. The potential of Factor H modulation to develop novel bio-pharmaceutical products to prevent, treat and control global diseases is detailed. Large drug companies are looking to devel- oping countries such as India for cost-effective clinical development of novel biotherapeutic drugs such as vaccines for HIV/AIDS. India offers the necessary legal and scientific framework for multinational compa- nies with product patents to develop novel biotherapeutic products cost-effectively. 565
    • Advances in Biopharmaceutical Technology in India Application of Recent Advances in immunology for Developing Novel Biotherapeutics in india 1. introduction I n translational development of biotherapeutic drugs, immunity is a major barrier.1 The human safety issue remains a problem that affects companies in both clinical trials and in post marketing phase.2-4 The drug industry’s problems associated with meeting the regulatory safety and efficacy standards for new products are analyzed in an FDA report on inno- vation stagnation published in 2004 (FDA).5 The cost of successful product from bench to bed-side has soared to US$1.7 billion by 2002. A growing list of fatal diseases compounds the problem of product pipe line and threatens the global consumers with next wave of morbidity and mortality from HIV/ AIDS, tuberculosis, malaria, SARS and bird-flu like fatal pandemics along with evolving threats of bio-terrorism. FDA estimates that if product safety can be predicted with 10% accuracy, it has potential to save up to US$100 million in new product development costs. The recent advances in the fundamentals of immunology have assigned a host protective role to Factor H.6, 7 Factor H is 150 kD immune regulatory protein of a billion year old early human defense system such as Alternate Complement System. The Alternate Complement System plays a crucial role in generating host inflammatory responses and is involved in education, instruction and priming adaptive or memory responses.8, 9 The potential of Factor H modulation to develop novel bio-phar- maceutical products to prevent, treat and control global diseases is detailed.10-12 2. immunity as a Barrier in translational Drug Development H istorically, the critical event in the development of modern biothera- peutic drugs was deciphering the double helix structure of DNA in 1953 by Watson and Crick. Since then, recombinant DNA tech- nology has given birth to numerous biotherapeutics such as recombinant insulin and erythropoietin. June 26, 2000, another milestone in genetics, is the day scientists announced the sequencing and assembling of the first rough draft –about 99% -of the genetic code that makes up the human genome. 566
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel BiotherapeuticsNovel biotherapeutic products incorporate living cells or subcomponents in thefinal product for the prevention, treatment and cure of diseases. This approachhas the potential to tackle a growing number of deadly diseases. Large drugcompanies are looking forward to developing countries for the cost-effectiveclinical development of novel biotherapeutic drugs such as vaccines for HIV/AIDS. TRIP implementation in India offers the necessary legal framework formultinational companies with product patents to cost-effectively develop novelbiotherapeutics.Biotherapeutic products such as vaccine, recombinant proteins and immunerelated drugs including monoclonal antibodies13 is class of drugs experiencingthe fastest rise in the new drug development process. Proteins are dynamicmolecules with the ability to adapt to various environment and physiologicalfactors. This flexibility allows a protein to transmit information from onebiochemical pathway to another. Changes to the shape of a protein can acti- tRiP implemen-vate, inactivate, or affect the function of a protein. The importance of sialic tation in indiaacid in the proper function of recombinant protein and antibody is increas- offers theingly recognized14-16 Recombinant human protein therapeutics; require necessary legalcomplex glycosylation, such as sialytation, for stability, solubility, folding, phar- framework formacokinetics, pharmacodynamics activity and to prevent host inflammatory multinationalreactions. Developing and marketing biotherapeutics based on recombinant companies withproteins and antibodies or its generic equivalents require specialized dedicated product patentsfacilitates of cell culture and fermentation technology. Manufacturing and to cost-effectivelymarketing of such products require different mind set and expertise than that is develop novelgenerally required for generic drugs. biotherapeutics.The biotherapeutics drugs such as recombinant proteins and monoclonalantibodies have added new layers of complexity and cost to the drug devel-opment process. For example, there are differences in the sialic acid bindingsites in apes and human. The differences in sialic acid binding between Apesand human beings apparently has contributed to the occurrence of cytokinestorm or “Acute inflammatory syndrome” in recently carried out human trialof antibody after it was proved safe in monkey studies. Six healthy young malevolunteers at a contract research organization were enrolled in the first phase1 clinical trial of TGN1412, a novel super agonist anti-CD 28 monoclonalantibody that directly stimulates T cells. Within 90 minutes after receiving asingle intravenous dose of the drug, all six volunteers had a systemic inflamma-tory response characterized by a rapid induction of proinflammatory cytokinesand accompanied by headache, myalgias, nausea, diarrhea, erythrema, vaso-dilatation and hypotension Within 12 to 16 hours after infusion, they becamecritically ill, with pulmonary infiltrates and lung injury, renal failure anddisseminated intravascular coagulation. After intensive hospital therapies all sixpatients survived.The full safety profile of a drug is rarely known at the time of approval by theFood and Drug Administration (FDA). Most drug-development programs are 567
    • Advances in Biopharmaceutical Technology in India designed for the treatment of symptomatic indications are under powered to detect any increased risk of rare drug reactions or change in background event rates attributable to the drug. Large, post marketing, randomized, controlled trials provide robust data on drug safety but are subject to multiple source of bias. Observational studies of a drug’s effects in clinical practice can offer addi- tional information on risks. Post-marketing observational studies permit the evaluation of drug safety in large number of patients in a real world setting, where practice patterns, including off label use of his drug can be assessed. Aprotinin was approved by FDA in 1993 as a mean of reducing periopera- tive blood loss in patients undergoing coronary artery bypass grafting. Neither the clinical trial database nor the numerous randomized controlled clinical trials conducted after approval identified an association between aprotinin and any short term increase in the risk of death or nonfatal cardiovascular or anyDue to the serious renal toxic effects.differences in Overwhelmingly, the drug development process involves two dimensionalsialic acid binding, models of in-vitro screening of chemotherapeutic drugs for efficacy againstthe successful pathogens and cancer cells. Such screening efforts do not take into accountresults from small the third dimension that is how the drug will behave once it is introduced inand large animal warm blooded animals. Immune response is different in small animals, largemodel often are animals and in human models. The drug interacts three dimensionally withnot translated host and its immune system most strongly in humans. Sialic acid binding is theinto human safety common point of interactions. Due to the differences in sialic acid binding, theand success. successful results from small and large animal model often are not translated into human safety and success. The host immune system reacts with endo- toxins, foreign proteins, antibodies and cells to induce inflammatory responses where sialic acid variations and its interactions with the alternate complement system are common denominators. Table 1 summarizes the role of immunity as a barrier in the translational development of drugs. 3. the Changing Role of Fundamentals of immunology: T he studies on immunity during the last few decades have mostly concentrated on the adaptive response in immunology and its hall- marks, that is, the generation of a large repertoire of antigen–recogni- tion receptors and immunological memory that relates to cellular and antibody responses. Current understandings of immunology are not complete and over the period of time have undergone numerous revisions 17-19 Therapeutic immune suppressive drugs developed and based on above understandings are extremely toxic, have many adverse effects and are associated with increased risk of infection and tumor potentials.20 Recent anthrax event prompted the preparation of a document titled “Stra- tegic Plan for Bio-defense Research” in 2002 by Anthony Fauci, Director of National Institution of Allergy and Infectious diseases (NIAID) branch of National Institute of Health (NIH) in USA ( 568
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel BiotherapeuticstABLE 1: immunity in drug development S NO PROPERtiES PHARMACEUtiCALS BiOtHERAPEUtiCALS NOVEL BiOtHERAPEUtiCS 1 Type of Drugs Synthetics Derived from live cell Combined with Live Cell or its Component 2 Molecular Size Small Large Variable EX: 1.Vaccine 1.Recombinant Proteins 2.Cell Transplant 2.Antibodies 3 Drug Discovery Structural Analogs Cell Culture System, IN-VIVO of synthetic Fermentation Combination compound Technology of cell or its By PhD (IN VITRO) Components scientist /chemist 4 Product short life Long Short sensitive Shorter to temp changes Sensitive to temp Changes 5 Adverse Effects Hapten based 1.Endotoxin 1.Endotoxin (Immune 2.Sialic acid difference 2.Sialic acid difference Related) 3.Early Immune reaction 3.Allo/xenogenic 4.Infection Risk 5.Early Immune Related 6.Delay Immune Related 6 COST <US$1 billion >US$1 billion >US$1.5 billionFrom a scientific and medical perspective, this plan is best seen as a variant ofthe general problem of emerging infectious diseases that have central char-acteristics of high morbidity and mortality due to inadequate diagnostic andtherapeutic interventions. The document highlighted the need to study threedimensional interactions of microorganisms with host and its immune system.It was acknowledged in the document that this is the field where experts lackedin-depth understandings. The plan emphasizes the importance of gaining abetter understandings in the three dimensional interactions of how pathogensinteract with the host and its immune system. The investments were justifiedby pointing out that such a research and development plan will have potentialspin off applications in immune related diseases of developed countries suchas in transplants and autoimmune diseases. The research will extend benefitsto immune related diseases of developing countries such as in their fightagainst Tuberculosis, HIV and Malaria. It was suggested that internationalcooperation with developing countries will facilitate the development of suchtherapeutics. 569
    • Advances in Biopharmaceutical Technology in India One of the major failing of the fundamentals of immunology is incomplete understandings as to how the immune system protects itself when acti- vated against foreign intruders. Currently scientists believe that the host is protected because of the presence of immune regulatory receptors in the cell membrane.21 In humans, immune regulatory proteins embedded in the host membrane are encoded by a cluster of genes located on the long arm of chromosome 1 (1Q32). This region is called as the regulator of complement activation (RCA) gene cluster. Although the proteins within the RCA family may vary in size, they share significant primary amino acid structure similari- ties. They are organized in tandem structural units termed as short consensus repeats (SCRs). The evolutionary relationships suggest membrane bound RCA proteins are functionally related. The recent advances in immunology have shifted attention to the working ofin humans, early human defense systems and the potential to educate, prime and developimmune memory based immune responses. Evolutionary immunologists date innateregulatory or natural defense system back 700 million years or more. The complementproteins system is an ancient microbial defense system. This is a system of fluid prote-embedded in the ases that interact with immune cells and its receptors to protect human beingshost membrane from ancient time against foreign intruders. The individual fluid proteases areare encoded numbered C1-C9. The complement system has three ways to activate andby a cluster of augment human defense system. One is antibody based that lead to the acti-genes located on vation of classical complement system. The second approach is lectin basedthe long arm of that is activated in response to presence of lectin based pathogen recogni-chromosome 1 tion molecules and third is through Alternate Complement system. As shown in Figure 1, the objective of three pathways of complement system, C3 is to(1Q32). generate common molecule C3. According to the recent advances in Immu- nology, the innate complement system differentiates self, and non-self, through generation of common molecule C3 and its active catalyzed product C3b. C3b is constantly generated in body fluid due to “enzyme tick-over hypoth- esis” by Alternate Complement System. If non-self or bacteria is identified because of carbohydrate signature it carries, C3b preferentially binds to Factor B to generate C3bB. This is further acted upon by Factor D to generate C3b convertase, C3bBb. Factor D is 25 kD immune activating circulatory protein of Alternate Complement System. It is mainly synthesized in adipose tissues. It also amplifies the activated responses of Classical and Lectin based comple- ment system. Factor H now regulate the complement activation by breaking down C3bBb to inactive C3bH. Active C3 fragmented products, C3a-C5a and C5b-C9, membrane attack complex have inflammatory components and stimulate the production of inflammatory cytokines. C5b-C9 in addition has cytotoxic component that form membrane attack complex and cause cytolysis. C5b-C9 cytotoxic immune responses are fluid counter part of cellular cyto- toxic immune responses initiated by Natural killer cells. Factor H also act as a controlling point in the amplifying cascade and is involved in inhibiting 570
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel Biotherapeutics C3 C3 C3b Factor B Ab C3bB NK cells B cell + Factor D C3bBb C5b-C9 CYTOTOXIC Th 1 cells Factor H Th 2 RESPONSES C3bH CD8T Factor I cells CD4T + IC3b C3a-C5a Phagocytosis CD4 Mature CD4 Immature Activation of Human Defence Responses + IC3b Breakdown Product Costimulate B CellsFigure 1: Recent advances in immunologyLegend: Figure 11. C3, three activating pathways of the complement system vis: Alternate, Lectin-based and Classical complement pathways2. C3, Common end product of 1.3. C3b, an activated C3 with thioester bond removed.4. Factor D, serine protease activiating protein of Alternate complement activating pathway5. C3bBb, C3 convertase that flag foreign pathogens6. Factor H, complement regulatory protein that has been assigned new role of recognizing self, host tissue as per recent advances in immunology. Microbes pirate this protein to mask its identity as self.7. C3a-C5a, activating intermediary complement products with host inflammatory actions.8. Macrophage, also CD 4 or immature dendritic cell in which entry of bacteria is facilitated through its surface receptors.9. CD 4 T matured dendritic cell where processed pathogen by 8 through phagocytosis express foreign signal by depositing peptide fragments in MHC 1 and MHC 11 groove10. CD 4 T cell of adaptive immunity that sense foreign signal according to “two Signal hypoth- esis”.11. Th 2 subtype of CD 4T cell that is involved in humeral cellular responses.12. Ab, antibody that is formed by stimulation of B cells by 11. 571
    • Advances in Biopharmaceutical Technology in India the formation of “Membrane attack complex” and at the same time inacti- vating C3b. The inactivated products of C3b such as iC3b, C3d, C3dg etc act as opsonin or natural adjuvant that attach breakdown products of foreign pathogen. Both inflammatory cytokines and natural adjuvant help in the maturation of dendritic cells by expression of co receptor function such as B 7 molecule. This according to “Two signal hypothesis” prime T cell responses. Another route by which iC3b and its breakdown product are involved in priming antibody responses is by acting as a natural adjuvant by binding to B cell receptors that amplify plasma cell function. Both T and B cell responses are involved in generating memory responses. In the absence of foreign pathogen, C3b binds with Factor H to form C3bH. Factor H mainly bind to sialic acid binding sites on the host cell surface. Factor H is a circulatory complement regulatory protein of Alternate Comple-Both t and B ment System. It has unique structure of twenty short consensus repeat (SCR)cell responses domain. Each SCR contain approximately 60 amino acids. Factor H is heavilyare involved glycosylated and has high sialic acid content. Factor H is the key self- recog-in generating nition protein that discriminates host –like features on microorganisms andmemory generate spectrum of activation rates for the complement system in differentresponses. microorganisms. A central question in innate immunity is how its various systems distinguish between potential targets and hosts. Factor H uses its 20 SCR domains to search for and interact with many ligands on a given target. Each SCR domain contributes to the recognition pattern, and if these sites work cooperatively in the groups of twos, threes and fours, etc, then by simple combinational math Factor H would have ability to discriminate among over 106 target surfaces. In the absence of foreign pathogen C3b does not form C3b convertase. The importance of recent advances in immunology emphasizing shift to early human defense system and is ability to educate, initiate and prime delayed adaptive immune responses is summarized in Table 2. The dominant role of circulating immune regulatory proteins to protect host cells against activated immune system is slowly being recognized. In evaluating the structure-function relationship of circulating immune regulatory proteins with membrane bound proteins, a study of genes involved in the regulation of complement activation (RCA) is particularly instructive. Both membrane bound and circulatory immune regulatory proteins are related. However, circu- latory immune regulatory immune proteins appear to be a dominant compo- nent in this relationship. 572
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel BiotherapeuticstABLE 2 Recent Advances in the Fundamentals of immunology Complement System Pathways S NO PROPERtiES CLASSiCAL LECtiN ALtERNAtE 1 RECOGNITION Ab-Complex MBL-Carbohydrate C3b-Microbe 2 COMMON MOLECULE C3 C3 C3 3 AMPLIFICATION ALTERNATE PATHWAY ALTERNATE PATHWAY ALTERNATE PATHWAY 4 REGULATION FACTOR H FACTOR H FACTOR H tYPE FUNCtiON SiGNiFiCANCE 5 EFFECTOR 1.C3b Convertase Tagging of foreign Pathogen particle- RESPONSES 2.C3a-C5a Chemo taxis Inflammation 3.C5b-C9b Cytotoxic Cell Lysis 4a.C3b CR1 Binding 4b.iC3b CR3/CR4 Binding 4c.C3d CR2 Binding 6 ADAPTIVE 2+3 of item 5 T Cell Responses Memory Responses RESPONSES 4a to 4c of item 5 B Cell Responses Memory Responses4. the Market Opportunity:I ndia is increasingly being promoted as an outsourcing destination for the development of biotherapeutic drugs. The advantages cited are lower costs, and shorter time duration for development of new drugs.22-23 TheUnited States market is the most matured and advanced in terms of patentlaws, drug development process, regulatory oversight to protect public safety,the market approval of drugs, and reimbursement systems. The US marketis therefore most lucrative for novel drugs. The US has a health care cost of$ 1 trillion in the year 2000. This cost will climb to $2.3 trillion by 2015.24TRIP implementation in the developing world coincides with the loss ofpatent protection on numerous drugs and biotechnology products in US. Thisprovides new incentives for drug companies in developing countries. India’smanufacturing and formulation experiences over the past three decades comesin handy for generic drugs where patents have expired. The cost of newdrug discovery and its clinical trials are soaring. The cost structure to developnew drug in developed world is very high. Multinational drug companies aretherefore moving to developing countries to lower their costs and to tap newmarkets. The drug companies in developing countries have tremendous incen-tives to rapidly adapt US based regulatory standards and business norms. Thecompliance by drug companies will facilitate their participation in developingand manufacturing generic and biogeneric drugs for developed markets. It 573
    • Advances in Biopharmaceutical Technology in India will help them to form collaborative partnerships with multinational drug companies and outsource their needs for new drug discovery, clinical trials, manufacturing and marketing. Developing countries such as India and China are experiencing rapid expansion of their health care markets. The consumer base in China and India covers 30 % or more of the global population. The ongoing trend to outsource medical care from the developed world to the developing world has accelerated. Both are adding new market opportunities in developing World. There are over 4000 biotechnology companies globally trying to compete to develop safe and effective products. By 2015, as TRIP implementation extends to under developed countries, the drug companies globally will compete for expanding market share. A critical insight or an improved understanding of human biology, particularly in the fundamentals of immunology for example at the molecular level havethe consumer the potential to improve safety of drugs in translational drug developmentbase in China and program and also has the potential to provide better solutions to diseases thatindia covers 30 affect global populations. In the modern drug development process immunity% or more of the is a common barrier for current and future therapeutics. As pointed out earlier,global population. circulating immune regulatory proteins such as Factor H bind to the poly anionic or sialic acid residues present on the host cells. Factor H along with membrane immune regulatory receptors neutralize activated immune proteins on the cell surface. Bacteria and viruses co-evolved with human existence. They constantly fight with human defense system for their survival advan- tages. The presence of sialic acid content in their cell membrane allows them to mask their identity as self. Many bacteria and viruses such as avian, feline, murine and simian RNA viruses are inactivated and lysed by human serum. In recent years, a large number of bacteria, viruses, parasites, and cancer cells are identified that pirate the functioning of human defense system by binding to the circulating immune regulatory protein or its downstream breakdown prod- ucts. This is due to their sialic acid content. Therapeutic modulation of Factor H and its potential to develop novel biotherapeutic drugs in different diseases is detailed as under. 5. therapeutic Applications of Factor H modulation A. Atypical Hemolytic Uremia Syndrome (aHUS): H emolytic Uremic Syndrome (HUS) is a severe disease frequently leading to end stage renal failure. Clinical features include rapid deterioration of renal function, anemia, low platelet count and frag- mented red blood cells in the blood. Current classification of HUS delineates two major types. Classical HUS occurs almost exclusively in childhood and is caused by bacteria releasing Shigela-like toxins. The disease starts with signs of enteritis, generally initiated by Escheria Coli strains, mainly 0157. Atypical HUS (aHUS) usually occurs in adults, and has been reported in association with a variety of conditions 574
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel Biotherapeuticssuch as therapeutic drug usage (Ovulation inhibitors and immunosuppres-sive drugs) diseases such as malignancies and systemic lupus erythramatosus),Pregnancy and after child birth. Familial occurrence of aHUS is reported insiblings and in a few families with autosomal dominant inheritance and rarelywith autosomal recessive transmission. According to one study of 111 patientswith aHUS, 14 % patients had FH1 germ line mutations.25, 26 To understandthe dominant relationship of circulatory immune proteins over membranebound immune regulatory proteins, it will be useful to consider what happensin the absence of Factor H. Congenital deficiency of Factor H or its mutation,leads to the activation of Alternate Complement System due to unopposedaction of Factor D. Since activated immune proteins are either not inactivatedor partially inactivated by membrane immune regulatory proteins, it leads to“Atypical hemolytic uremia syndrome” (aHUS). The therapy at clinical level isto replace Factor H that is mainly synthesized in liver. An alternate approach there is ais to inhibit activated complement system such as by modulation of Factor H. perception thatThus Factor H modulation could be life saving in these patients. successful drugB. HIV/AIDS Therapeutics: companies withi. The Role of India: There is a perception that successful drug companies novel productswith novel products such as an HIV vaccine must balance ethics and morality such as an HiVin business practices, and issues of profiting from deadly yet preventable and vaccine musttreatable diseases such as HIV must be considered. This has played a role in balance ethicsencouraging India generic drug companies to take advantage of the Indian and moralitypatent laws of the 1970s that allowed them to meet growing public demand in businessfor cost-effective medicines. It is a surprise to many people including Indian practices…government officials that it is estimated there are close to 6 million HIVpatients in India and. HIV is expected to grow four folds to 25 M by 2025.There is an added risk that 50 % of these patients will have reactivated tuber-culosis that may be resistant to currently available drugs. Since half of HIVpatients in developing countries may harbor tuberculosis that may be resis-tant to multiple drugs, the future scenario of public health in India is a chal-lenge. In March 2005, the Indian parliament passed the third amendment tothe Indian Patent Act (1970) that led to the introduction of “Product Patentregime”. This has raised serious questions regarding the role of generic drugindustry and its ability to compete and at the same time provide the continuingavailability of essential medicines at affordable prices.ii. The Dilemma of Brand vs. Generic Drugs: New drug developmentin developed countries is costly. It is a high risk venture that takes 10-15 yearsof sustained effort. The chances of success is only 30 % after animal tests arecompleted. In 20th century the traditional pharmaceutical industry played adominant role in the control of major infectious and chronic diseases. Thisreduced greatly the morbidity and mortality for example HIV/AIDS in devel-oped countries. The high charges for new medicines by major pharmaceuticalcompanies were justified in developed countries. The medical management 575
    • Advances in Biopharmaceutical Technology in India of patients in developed countries is supported by adequate reimbursements for new technology and patented drugs. Perhaps, the high reimbursement rate and profits in developed country could be used effectively to innovate at marketing level to lower the cost of drugs through outsourcing. However, high cost drug models in developing and under developed countries is problematic. There are no adequate laws to protect patented inventions. There is no reim- bursement mechanism for marketed patented products. The ongoing drama of affordability of essential medicines for example in HIV/AIDS is played out in major news papers. In the 1970s India introduced product process patent laws. The laws were mainly geared to stimulating competitive capabilities of local drug industries. It was an enormously frustrating experience at the regulatory level to introduce advanced products based on Western patents. Indian patent laws of the 1970s[t]he indian provided a way for Indian drug companies to do what was legally allowed:drug companies copy patented products by bypassing the manufacturing and drug developmentreduced the process. From a regulatory point of view the generic drug company only needscost of drug to prove that their products are bioequivalent to patented drugs. Thereby, thedevelopment Indian drug companies reduced the cost of drug development to 1/1000th ofto 1/1000th of patented cost. This allowed the Indian drug industry to flourish in developingpatented cost. and third world countries. It allowed them to compete with multinational drug companies effectively. Copied products were marketed at a fraction of the cost that was charged by multinational companies. This benefited consumers to meet their dire needs for products that were safe, low cost and quality based. In the public eyes generic drug companies were perceived as benefactors of humanity and multinational drug companies were branded as exploiters of human rights. Despite scientific progress and therapeutic advances –cost ineffective medicines and cheap generic counterparts have proliferated and the demand –supply gap for truly life saving drugs have widened. This is a true international emergency of the 21st Century. Leaders around the world are scrambling to find mean- ingful solutions to contain the rapid global spread of infectious disease such as HIV/AIDS. iii. The Adverse Impact of Patent laws of 1970s: The patent laws of the 1970s included controversial Section 5(1) which provided for process patents and stifled innovation in India. There is no market mechanism to adequately reimburse patented products. The invention and discovery of new drug development is considered costly, economically not viable, and wasteful of time, money and energy by academic centers, financial centers and experts in India. This inertia is reflected in poor venture capital investment in research and development of new drugs. The hospital system, the education system and medical industry in general remained apathetic and negligent in their mission objectives to serve humanity. Availability of cheap drugs and lack of appro- priate reimbursement mechanisms or insurance system led to the neglect of 576
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel Biotherapeuticsmedical illnesses and its follow up in developing countries. This has put Indiainnovation several years behind developed countries.iv. New Patent regime in India: The Indian parliament’s third amend-ment to the Indian Patent Act (1970) that led to the introduction of “ProductPatent regime” raised serious questions regarding the role of generic drugindustry and its ability to compete and at the same time provide the continuingavailability of essential medicines at affordable prices. The recent introduc-tion of Trade related intellectual Property Agreements (TRIP) is intended tocorrect the past imbalance by creating level Plainfield where an individual or asmall company can compete with giant drug companies based on intellectualproperties.In the US, the patent process is governed by ’Prior User’s right’ while inEurope it is governed by ’First to file’. The typical requirement for an inventionpatented in the US is that it has to be useful, new and not obvious.27India have adapted Trade Related Intellectual Property Agreements (TRIPs)and raised the bar for patented inventions to avoid spurious claims. In orderto develop successful patent under section 2 (i) (j) of Indian patent act, a newdefinition for “new invention” has been added. ‘New invention’ is defined as,“any invention or technology which has not been anticipated by the publica-tion in any document or used in the country or else where in the world beforethe date of filing of the application with complete specification, i.e. the subjectmatter has not fallen in public domain or that it does not form part of the stateof art”. Patents and Gleevec: Gleevec (spelled Glivac outside the United States) is used to treat chronic myeloid leukemia. This drug has been patented by a multinational drug company in 35 countries and is sold for $26,000/ year. The drug was rejected in India under section 3(d) and is a subject of ongoing legal battle. Gleevac, the brand name version of imatinab mesylate is offered by generic drug companies at 1/10th price by obviating the need for clinical trials. (28). Under the amended section 3(d) of Indian patent act the mere discovery of new use, new property, use of a known process, machine or apparatus or combination of two products is not patentable if it does not result in new product with increased efficacy of known compound. According to current Indian patent laws not patentable clause under section 3 (i) include “any process for the medicinal, surgical, curative, prophylactic, diagnostic or therapeutic or other treatment of human beings or any process for a similar treatment of animals to render them free of disease or to increase their economic value or that of products.” 577
    • Advances in Biopharmaceutical Technology in India With reference to the inventive step, the Indian Patent Act states: “‘Inventive step’ means a feature of an invention that involves technical advance as compared to existing knowledge or having economic significance or both that makes the invention not obvious to a person skilled in the art”. The off label use of the drug can be patented as method of treatment in devel- oped countries. It is stated that medical –process patents threaten to complicate medical practice, increase cost, and restrict access to new procedures. Owing to such concerns, nearly 80 countries refuse to grant patents on medical proce- dure.29 Adaptation of US patent laws of ’Prior user right’ as opposed to ‘First to file’ European and Indian patent laws have the potential to remove abusive practices and may contribute to healthy industry competition. The patient is benefited by having access to needed drugs and the industry experiences healthier competition. The outcome of the implementation of TRIP laws in[P]atients with India is that in August and September 2006, patients with cancer, lawyers forcancer, lawyers patient advocacy groups, and representatives of nongovernmental organiza-for patient tions (NGOs) converged on the offices of a multinational drug company inadvocacy Mumbai, India, to protest the company’s efforts to obtain an Indian patent.groups, and This situation where novel applications of an established drug, in a new formu-representatives of lation and in new devices is of growing importance with the rapidly risingnongovernmental cost of drug development in resource limited country such as India. There isorganizations an urgent need to develop innovative and new solutions at the clinical level.(NGOs) converged Although, most physicians are allowed to use established drugs for new appli-on the offices of cations, in a given patient the safety and efficacy of the off label drug usea multinational remains questionable. If an off label drug in a new formulation is effective indrug company in a fatal disease – there is nothing wrong in patenting the drug and it should beMumbai, india, welcome. From regulatory perspectives, the drug authorities have taken a standto protest the to allow the use of such drugs only after adequate clinical trials. This has a fixed clinical trial cost. The off label uses of the drug for new disease applica-company’s efforts tions either in existing or new formulation is a commercial activity. Anotherto obtain an important step taken by US FDA is to punish the drug company with severeindian patent. penalties for promoting the off-label use of drugs to physicians. Since clinical trials of a drug itself is a costly enterprise, there is very little incentive on the part of physicians or the drug industry to undertake large clinical trials to assess the importance of existing drugs in new deadly diseases unless they can patent it and prevent competition from generic companies. Patenting of new applications of existing drugs in new formulations is the fastest way to generate new therapeutic drugs at cost effective prices. It obviates the need for manu- facturing that can be outsourced. This can result in saving of US$200 million in up front cost. According to the US FDA, if the safety of the drug is known or if it can be predictable by 10 % it has potential to save US$100 million in drug development costs. The existing drug in new formulations has predictive potential of safety by at least 50 %. This should reduce the cost of drug devel- opment by US$500 million. 578
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel BiotherapeuticsIn the era of new patent regimes, the mindset of Indian drug companies andgovernment organization is tuned to serve the need of multinational drugcompanies as outsourcing partner and offset the impact of TRIP by findingnew opportunities in the developed world. A large number of patented drugsthat have annual sales of over US$100 billion in the US face expiry of patents.This is a new opportunity for Indian drug companies to compete internation-ally in developed countries for the generic drug market. Implementation ofTRIPs coincides with the global explosion of knowledge. India has provenexpertise in the management of information technology and its evolving tools.It is this leadership that is combined with patented technology that is helpingIndian companies to handle knowledge base at global level. This is defining thenew levels of competition and cooperation to gain new leadership role in thefight against global diseases.v. Factor H Modulation for HIV Therapy: Responding to the rising A large numberproblem of drug resistance, experts have opined for the need for better, simpler of patentedand safer drugs that target the HIV genomic structure in a novel way and at drugs that havemultiple points so that the risk of resistance obtained with current antiretro- annual sales ofviral drugs could be reduced.30 Another leading expert in immunology has over US$100commented that in view of the limited therapeutic efficacy of current Highly billion in the USActive Anti Retroviral Therapy (HAART) regimes, there is an urgent need face expiry ofto combine such therapy with immune based approaches that may improve patents. this is atherapeutic efficacy of cocktail therapies.31 The modern drug discovery begins new opportunitywith genomics and proteomics. Thus for example the role of proteomics is for indian drugevaluated to identify HIV-1 therapeutic targets in host proteome. Identification companies toof novel targets for therapy that may not develop resistance is sorely needed. compete…Therefore, the effects of Tat protein on cellular gene expression during variousphases of the cell cycle of HIV replication have been studied. This has led toidentification of several host proteins including Factor H that may offer noveltherapeutic target.32 In the basic science literature the in-vitro experimentsdone with serum of HIV patients has shown that Factor H modulation killHIV outside the immune cells.33 The recent advances in the fundamentalsof immunology have identified the growing importance of Factor H in hostprotection. Factor H is an immune regulatory protein of Ancient humandefense system, Alternate Complement System. Factor H is pirated by HIVwhile simultaneously it also activates complement system. The combina-tion of piracy of Factor H and activation of complement system allows HIVto neutralize and prevent the generation of fluid based cytotoxic immuneresponses. The breakdown products of immune activation play an importantrole in persistence of HIV infection in HIV treated patients and contributeto future resistance pattern.34, 35 Factor H modulation therefore has potentialto prevent, treat and cure HIV. 36 However, the translational development ofan antibody therapy is problematic. Differences in sialic acid between apesand human beings may precipitate ’acute inflammatory syndrome.’ Similar 579
    • Advances in Biopharmaceutical Technology in India syndrome may also be precipitated with the development of Factor H anti- body. The congenital deficiency of Factor H leads to Atypical acute hemolytic uremia syndrome (aHUS) characterized by hemolysis, hypotension and renal failure. A novel therapeutic approach that inhibit Factor H while regulating the activation of Factor D is urgently needed to overcome above risk and enhance safety of novel biotherapeutic products. Figure 2A details how HIV pirates the functioning of human defense system. HIV is an enveloped virus and has allo MHC 1 and MHC 2 proteins of the primary host as well as the immune regulatory machinery on its surface membrane. Once HIV enters the host body, it is efficiently sensed by comple- ment system. Both Classical and lectin based complement system is activated. This effort is amplified by Alternate complement system to generate C3b Convertase, 11, 12 and its down stream products such as C3a-C5a and C5b to C9. However, HIV simultaneously binds to circulating Factor H on its surface to form C3bH-HIV, 13 that inactivate C3b Convertase on its surface to prevent C5b-C9 cytotoxic immune response. The exact details of HIV binding site to Factor H has been detailed and appear to be on gp 120 as well as on gp 41. C3a-C5a draws immune cells to inflammatory sites to cause host damage, 14. The inactivated C3b products are used as natural opsonin to bind to HIV particle. Inflammatory milieu is used to facilitate HIV entry inside CD 4 cells. HIV multiplies and repeats its cycle of freshly infecting new immune cells. Inactivated C3 products such as iC3b and its breakdown products along with C3a-C5a that normally prime dendritic cell maturation is circumvented by HIV. HIV uses complement breakdown products as opsonins and use comple- ment receptors on the cell surface also to shuttle to follicular dendritic cells and persist there. This is the underlying mechanism for the failure of HAART regime. The crucial role played by Factor H in protecting HIV is evident if HIV is incubated in Factor H depleted sera from HIV patients. The absence of Factor H leads antibodies of HIV to sense the presence of HIV as foreign molecule and leads to prompt cytotoxic immune response to kill HIV outside the cell in the serum with 80% efficiency. Use of membrane bound immune regulatory Antibody, however brings only marginal improvements. Several new advances in the understanding of microbial interaction with Factor H have occurred. One critical problem is how one will control the prevention of atypical hemolytic uremia. One approach is controlled manipulation of Factor H and Factor D by developing a fusion molecule. Such an approach will lead to inhibit Factor H while preventing the over activation of Factor D. Dual strategy should work well to improve current methods of HAAART therapy. One potential advantage is by targeting immune evasion mechanism, the future risk of mutation and potential outbreak of new viruses could be prevented. As shown in Figure 2B, HIV interacts with Factor D, Factor H and CD 4 T cells. Therapeutic strategy aimed to control Factor D will reduce host related 580
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel Biotherapeutics Self C3 Non-self (Level of Hierarchy) Factor H Factor B C3bH-HIV C3b C3bB 11 Factor D Aborted C3bBb 12 C3bH-HIV New HIV 13 Host Damage 14 DC Immature 15 CD 4 Cell DC Mature Aborted CD 4 Cell T Cell Memory Aborted ResponseFigure 2A: HiV piracy of Factor H C3 C3 Ab C3b B cell Factor D C3bBb HIV Th 2 Factor H C3a-C5a CD4T CD4 Mature Host In ammation CD4 ImmatureFigure 2B: HiV piracy of Factor H 581
    • Advances in Biopharmaceutical Technology in India inflammatory damage by HIV and will prevent its persistence in follicular dendritic cells. This strategy will improve outcome of HAART regime. The persistence of HIV and the risk of resistance can be reduced. An alternative strategy is to inhibit Factor H to stimulate Alternate complement system. Since in blood of HIV patients there are already circulating Antibody, removal of Factor H will unmask the identity of HIV as foreign pathogen. Circulating Antibody will initiate cytotoxic immune responses to kill HIV. Incorporation of this strategy in most modern vaccines will lead to expedient solution to HIV/ AIDS crisis. Figure 3 shows the interaction of HIV with Factor D and Factor H is outside the CD 4 cells. Thus Factor H modulation will kill HIV outside the cells while protecting host from undue activation of Factor D. Figure 3 582
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel Biotherapeutics5 C. HIV Vaccine and its Availability:Historically, vaccination is the best method of preventing infection.37, 38 Amongthe difficulties confronting researchers are viral heterogeneity, the lack ofpractical animal model and ethical dilemmas involved in conducting primaryprevention trials in the United States and abroad. India is being promoted asan outsourcing destination to develop novel biotherapeutic drugs. Interna-tional Association of Vaccine (IAVI) has spearheaded the project to do HIVvaccine trials in India. A large number of drug companies have responded tothis call and are heading to India to initiate vaccine trials. HIV/AIDS vaccineis a reference example of novel bio-therapeutic product that includes a compo-nent of viral particle. The critical problems are the ethics, human rights andneed to protect volunteers. HIV has sialic acid binding sites. This bindingsite is in the cell membrane and is located at gp120/41. This apparently hasresulted in the difference in simian virus and HIV in human beings. Thus for india is beingexample, Simian virus doesn’t progress to AIDS in monkeys. The vaccine for promoted asHIV developed and tested in monkey model has yet to prove its clinical merit an outsourcingin human trials. In human beings HIV readily progress to AIDS. The vaccine destination todata generated in monkeys therefore is not predictive of its capability to block develop novelor prevent progression of HIV to AIDS. Most modern HIV vaccine efforts are biotherapeuticgenerated to develop vaccine responses that target Gp 120/41. However, due Factor H binding, they are not able to generate antibody responses. Thisis the reason for current failure of HIV Vaccine to induce cytotoxic immuneresponses.The crucial role played by Factor H in protecting HIV is evident if HIV isincubated in Factor H depleted sera from HIV patients. The absence of FactorH leads antibodies of HIV to sense the presence of HIV as foreign moleculeand leads to prompt cytotoxic immune response to kill HIV outside the cell inthe serum with 80% efficiency. Use of membrane bound immune regulatoryAntibody, however brings only marginal improvements. One potential advan-tage is by targeting immune evasion mechanism, the future risk of mutationand potential outbreak of new viruses could be prevented. On the basis of thisdata, Factor H modulation is considered by experts as novel “Complemen-tary AIDS Vaccine.39, 40 Since in HIV vaccine trials ethics is a major concern,Factor H modulation is a stand alone HIV Vaccine approach where there is noHIV particle is incorporated.D. Age-related macular disease:One additional approach to drug discovery is information- Information ofspecific genes and its related proteins. These tools begin with bioinformaticsand exploration of public gene bank data sites. To create a protein, messengerRNA (mRNA) – a copy of one DNA strand of a gene – is sent out of thecell’s nucleus and into cytoplasm, where protein is synthesized by ribosomethat recognize and link to gather specific amino acid chains that make up aprotein. Single Nucleotide Polymorphism (SNP) is a technique to study genes 583
    • Advances in Biopharmaceutical Technology in India and its variations in amino acid and mutation. In the spectrum of blindness, few rival Age-related Macular Disease (AMD). With aging population, AMD is rapidly identified as novel area for drug development by multinational compa- nies.41 Modern drug discovery efforts have expanded to take advantage of new discovery tools of genomics.42-44 Macula is the portion of retina that perceives the sharpest external image. With aging, the cells of macula undergo degen- erative changes. The whole mark of AMD is an inflammatory lesion called as Drusen’s lesion. Several drug companies have products under development to inhibit the molecular target identified as angiogenesis. However, therapeutic manipulation of this target is applicable to less than 10% of advanced cases limiting therapeutic efficacy and market size. Several large scale trials have found that in 50% or more cases of AMD, Factor H is mutated leading to the activation of Alternate Complement System.the globalization E. The Cure of Type 1 Diabetes:trends along with The use of recombinant insulin, its novel formulations and newer better oralrising standards hypoglycemic agents have improved therapeutic results in the management ofis creating eating diabetic patients. However, these efforts have not cured diabetes. It is not onlypatterns that is a disease of developed countries but also a major problem in developing coun-not healthy and tries. The globalization trends along with rising standards is creating eatingcontributing to patterns that is not healthy and contributing to obesity, diabetes and increasedobesity, diabetes cardiovascular diseases.45 First generation of biotechnology companies success-and increased fully manufactured recombinant insulin and its varied formulations are nowcardiovascular available in clinical practice. The cure of diabetes by pancreas transplant isdiseases. now an accepted clinical procedure and has success rate of 80-90% in the first year while by five year it declines to 57%.46 A successful cure of diabetes and proof of concept was provided for this method in 1993 by using biocompat- ible membrane that is substantially free from endotoxin content.47 Current methods to cure of diabetes require that biocompatible material used to encapsulate insulin producing cells must be substantially free from endotoxin content. Advances in immunology are leading to the better understandings of the immune rejection process. In spite of these advances, the currently prac- ticed technology for the cure of diabetes has many flows and requires further improvements. In 400 plus patients transplanted the success rate of cure of diabetes drops from 50% in first year to 33% by third year and to less than 10% by 5th year. Insulin producing cells get immune rejected. Prior to the year 2000 the yield of insulin producing cells was less than 10 % per pancreas and required as much as 5.6 pancreas to cure one patient with Type 1 Diabetes. By 2006, the technology has been improved, reducing the need for pancreas from 5.8 to one.48 A further technology advancement involving stem cells to manufacture insulin producing cells will outdate the need for donor pancreas in next few years.49 Such an advance will allow one to cure Type 1 diabetes at an earlier stage and in a broader level. It is estimated that in the USA total supply of pancreas is 6000 per year. Since, the cure of diabetes by single 584
    • CHAPTER 14 n Application of Recent Advances in Immunology for Developing Novel Biotherapeuticspancreas in 2006, is reported, it is expected that number of islet cell transplantwill rise dramatically in coming decade. Economics of each cure of diabetescost approximately US$150,000/year that has current success rate of 50% atone year but drops to 10% by 5th year.50 Technology improvements may openup a huge market opportunity. With organ donation laws in place in India,where no islet cell transplants are performed, this provides an opportunity tocollaborate and cooperate. The model example could be extended to clinicaltrials of other diseases. Collaborative sharing of data and open access to trialdata, will speed the introduction of new biotherapeutics, identify early deadlycomplications and adverse effects and provide a mechanism of accelerating theclearance of regulatory hurdles.F. Therapy for Renal Diseases:In 1995, Institute of kidney diseases in Ahmedabad, GS. initiated “Cell the institute ofTransplant Laboratory for the cure of diabetes.” However, due to difficultiesinvolved in curing diabetes, they shifted attention to developing stem cell tech- Kidney diseases,nologies to address immune rejection problems in kidney transplant patients. incidentallyThe Institute of Kidney diseases, incidentally is a largest transplantation is a largestcenter in Asia offering low cost renal transplants. The key problem identified transplantationin kidney transplant patients has been the development of memory responses center in Asialeading to immune rejection and inadequate response to immune suppressive offering low costdrugs. The Institute of Kidney Diseases has shown their stem cell technology renal transplants.advances and how CD 34 cells from donor bone marrow could be transplantedinto recipient patients.51 This procedure helped to reduce cost and effec-tively reduced the need for immune rejection drugs leading to the successfuloutcome in transplanted patients. Many of the transplanted patients however,were related donors where immune system responses are likely to be weaker.The institute presented several cases of renal diseases as a part of continuingmedical education. New advances in the fundamentals of immunology havethrown new lights on the cause and occurrences of adverse effects. This willlead to better safe immune suppressive drugs with least adverse effects.52G. Cancer Metastasis:The human immune system is designed to defeat disease and has the abilityto combat cancer cells. Recent advances of immunology explain and providenew insight into how cancer cells evade immune responses.53 A large numberof cancers such as breast, bladder, cervix, lung, prostate, ovarian, tropho-blastic tumors, and thyroid and glioblastoma multiformes are identified thatsecrete Factor H or pirate its functioning.54-57) The piracy of Factor H allowscancer cells to protect against complement mediated formation of MembraneAttack Complex (MAC). Cancer cells may either prevent the formation of C3convertase or inactivate MAC on its surface. Therapeutic implications are thatincorporation of Factor H modulator can be exploited strategically to generatevaccine responses in cancers or to provide immunotherapy. 585
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    • About Advances BiopharmaceuticalTechnology in IndiaT he biopharmaceutical industry in India has grown dramatically over the past few years, and sales have exceeded US$1.5 billion. This study describes the Indianbiopharmaceutical industry, its history, its advantages and opportunities, as well asits challenges and risks. Today, the biopharmaceutical industry in India has broughtseveral protein drugs to market and is developing many more. The next several yearswill be interesting as India takes its place on the global stage. Biopharmaceuticalproducts have a long history in India, and trace their roots back several thousandyears through schools of healing practice. The Indian government is currentlyworking towards developing that experience into a sound biotech industry. Thecountry’s objective is to help minimize foreign dependence, especially in high-techareas. This study describes the industry’s history, and the Indian government policiesthat have helped enable the manufacture of modern biotech products at affordableprices. We discuss the patent factors and history that have shaped the Indian industry,including the industry’s reliance on production of outside-of-patent products. As theIndian government continues its efforts to create alliances between private industry andresearch institutes, the next decade should show a significant growth in the Indian biotechindustry, and novel biotech drugs may ultimately dominate. India is expected to emergeas a strong player in the production and sale of biotech products in the coming years, aslocal consumption rises, and as its local biotech industry takes steps to develop a globallycompetitive local industry that stands on a foundation of basic research. Authors from:  M J Biopharm Pvt. Ltd. Endocrine Technology, LLC GAEA Resources Inc. Bharat Biotech International Ltd. Magnetical Finston Consulting, LLC neo BioMed Services CanBiotech Chiltern International Private Ltd. beòcarta Ltd. Biospectrum Asia Health Interactions Chiltern International Pvt. Ltd. Serum Institute of India Ltd. national Institute of Health Jim Schnabel, BioPlan Associates, Cadila Pharmaceuticals Ltd. Uniformed Services University of the Inc. J. Sagar Associates Health Sciences John Curling Consulting AB Amgen, Inc. national Institute of Science ProMetic BioTherapeutics, Inc. Technology and development Apothecaries Ltd. Studies Celestial Biologicals Ltd Chiltern International Private Limited dow AgroSciences LLC Strategic Manufacturing Worldwide, Inc. ISBN 978-1-934106-07-5 9 781934 106075