P. Pushpangadan National Botanical Research Institute

1. Biodiversity Wealth and Opportunities for Asia Pacific Countries in Biotechnology & Herbal Technology P. Pushpangadan National Botanical Research Institute (Council of Scientific &Industrial Research), Rana Pratap Marg, Lucknow-226001 (September 2003)

2. WORLD TRADE AND ECONOMIES: THE PARADIGM SHIFT • “Resource – based economies’ to ‘Knowledge –based economies’ • 21 st Century will be the Century of Knowledge”…… “A nation’s ability to convert knowledge in to wealth and social good through the process of innovation will determine its future” ( R A Mashelkar, 2001)

3. 21st Century 21st century is the century of Biology powered and propelled by scientific knowledge and technological expertise • Three technologies namely • Biotechnology • Herbal technology • Information technology (Bioinformatics) • are going to be the most powerful elements that are crucial for prosperity and welfare for the people of nations.

4. Herbal technology All technologies for the manufacture of value added plant products can be called as herbal technology • Herbal drugs and pharmaceuticals, • Nutraceuticals, • Functional foods, designer foods or health foods and health drinks • Cosmaceuticals • Biocontrol agents • Biopesticides

5. WORLD TRADE AND ECONOMIES: THE PARADIGM SHIFT • “Resource – based economies’ to ‘Knowledge –based economies’ • 21 st Century will be the Century of Knowledge”…… “A nation’s ability to convert knowledge in to wealth and social good through the process of innovation will determine its future” ( R A Mashelkar, 2001)

6. Genesis of the Global Concern on Biodiversity Conservation • UNEP constituted an ad-hoc Working Group of Technological and Legal experts to prepare an international legal instrument for conservation and sustainable use of Biodiversity which resulted in ‘CONVENTION ON BIOLOGICAL DIVERSITY’ (CBD). • 171 countries signed CBD in June 1992 during the Earth summit at Rio de Janeiro. • CBD came - into force as an International Law on 29th Dec. 1993. • 186 countries are now parties to CBD (as on Feb. 2003)

7. Conservation of Biodiversity Strategies & Priorities IUCN, UNEP & WWF 1980 came out with the first Global Strategy for Conservation. This Strategy defined conservation as: “Management of human use of biodiversity so that it may yield the greatest sustainable benefit to present generation while maintaining its potential to meet the needs and aspirations of future generation” This definition involves two complementary components ‘Conservation’ and ‘sustainability’

8. Biodiversity & TK: Capital Assets of Asia-Pacific Countries • Biodiversity and TK are two invaluable capital assets of South countries for: • Building up IPR- covered bio-industrial enterprises • Herbal Drugs • Pharmaceuticals • Natural product development sectors • Generating economic wealth and improving quality of life and well- being of people Contd…….

9. Biodiversity & TK: Capital Assets of Asia-Pacific Countries (Contd.) • Building up S&T capability in advanced technologies of bioprospecting • Human resource development in Biotechnology, Bioinformatics and Bioprospecting • Empowering local and indigenous communities for conservation, sustainable use and building up location –specific biodiversity enterprises through S&T intervention

10. Biodiversity & TK: Bridging the North-South Gap • Asia-Pacific countries SHOULD: • Develop capability in biotechnology, bioinformatics and bioprospecting through national, regional and global ‘biopartnership’ programmes • Address and resolve the issues of access to and transfer of genetic resources and technologies between North- South countries Contd……..

11. Biodiversity & TK: Bridging the North- South Gap Asia-Pacific Countries should: • Prevent bio-piracy and misappropriation of genetic resources and TK • Develop an international system for protection of TK • Develop effective mechanisms for fair and equitable benefit sharing and technology transfers

12. Convention of Biodiversity (CBD)Relevant Provisions of CBD • Article 3 recognizes the sovereign rights of States over their biological resources. • Article 15 states that when access to genetic resources is granted, it shall be on mutually agreed upon terms and subject to Prior Informed Consent. • Incentives to biodiversity-rich countries to conserve and sustainably use their genetic resources, including joint research, access to & transfer of technology (Articles 15,16).

13. Relevant Provisions of CBD(Contd...) • Article 16.2 addresses issues surrounding the access to and transfer of technology, governed by patents. • Article 16.5 anticipates the difference in objectives between IPR regimes and the CBD and seeks to ensure that IPRs don't run counter to the CBD. • Article 8(j) underlines the need to protect TK and points to the potential benefits to be realized from such knowledge through involvement of its holders and equitable benefit-sharing.

14. Relevant Provisions of TRIPs on Biological Resources • Under Article 27, virtually all inventions are to be patented if they are new, involve an innovative/inventive step and are capable of industrial application. • Exceptions to patentability include plants, animals ( other than microbes) and biological processes for the production of the above. However plant varieties must be protected either by sui generis or by patenting (27.3(b)).

15. Relevant Provisions of TRIPs on Biological Resources(Contd….) • Article 30 confers limited exceptions to the rights conferred on patent holders, taking into account the legitimate interests of third parties. • Article 29 imposes two conditions on patent applicants; that they disclose the invention clearly and completely enough for a person skilled in the art to reproduce it and it 'may' require an applicant to provide information concerning the applicant's corresponding foreign applications and grants.

16. TRIPS-CBD Relationship • Absence of explicit compatibility, Difference of approach and priority given to issues which are ultimately related. This has led to violation of the CBD (Articles 8,15 &16). • TRIPs ignores a vast range of valuable, traditional knowledge (TK) because it doesn't meet the standards of patentability.

17. TRIPS-CBD Relationship(Contd..) • TRIPs undermines CBD in cases of biopiracy, by putting the burden of proof on the source country rather than patentee. Identification of unique source material as required in Art.29 of TRIPs is insufficient. Lack of transparency in the patent application procedure. • TRIPs doesn't require the recognition of domestic laws protecting access to genetic resources and TK and subsequent benefit sharing.

18. The Need to Amend the TRIPs Agreement • Absence of a clear reference to CBD and the relationship with CBD could hinder the implementation of the latter by violating the primary principle of sovereignty over genetic resources. • TRIPs should provide international recognition of relevant domestic legislation of its member countries, especially as far as access and benefit sharing issues are concerned.

19. The Need to Amend the TRIPs Agreement (Contd...) • It is far more cost effective in the long run to establish an internationally accepted solution through TRIPs for the prevention of biopiracy. • TK associated patents have fetched large profits and it makes ethical and economic sense for TRIPs Agreement to recognize a need for benefit sharing.

20. The Need to Amend the TRIPs Agreement (Contd...) • It is far more cost effective in the long run to establish an internationally accepted solution through TRIPs for the prevention of biopiracy. • TK associated patents have fetched large profits and it makes ethical and economic sense for TRIPs Agreement to recognize a need for benefit sharing.

21. Bioprospecting and the new IPR regime Given the global trends in capturing the intellectual property markets, the Third World nations in the Asia pacific now needs to look ahead for the best possible ways and means by which they can generate IPR and build up IPR covered bioindustrial regimes. Biotechnology (BT), Information Technology (IT) and Herbal Technology (HT) are the three fast emerging and powerful areas of R&D in current century. The rich biodiversity, associated knowledge systems and human resources etc. are the strength of Asia-pacific countries, and therefore have the best opportunity.

22. Bioprospecting • Gene Prospecting • Genetic engineering • Crop development • Fermentation • Cell culture • Chemical Prospecting • Drugs and pharmaceuticals • Pesticides • Cosmetics • Food additives • Other industrially valuable • Chemical products • Bionic Prospecting • Designs • Sensor technologies • Architecture • Bioengineering • Biomodeling

23. Bioprospecting: Linkages and leads Biodiversity & IK/TK Biotechnology Bioprospecting Information technology • Drug development • Pharmaceuticals • Agrochemistry • Cosmetics • Proteins • Enzymes • New crop varieties • GMOs • GM foods • Designs etc. Herbal technology Conservation Sustainable use Benefit sharing Bioinformatics IPR

24. IPR Issues / Benefit Sharing Strategies • Appropriate Procedures for IPR Protection/Benefit sharing • Documentation & Registration of TK – Medicinal plant use & Conservation at local , state and national level. • Contribution to TKDL & TKRC • Value addition to TK & Indigenous Medicinal Plants – Scaling up IPRs • Herbal drugs, Pharmaceuticals, Natural products & byproducts, Nutraceuticals, Functional foods, etc.

25. Some features of international health care • Health care policies largely market driven by the pharmaceutical industry diverting attention from health preservation to illness cure • Prevention and eradication of diseases undermines the economic basis of this industry • No satisfactory drugs available for most of the degenerative disorders characteristic of graying population and for re-emerging resistant infections • Many currently used modern drugs do not have valid proven clinical utility. • USA has among the highest per capita annual expenditure on health care ($3600) but still about 15% population is denied even basic care facilities (the best Indian state like Kerala have an annual per capita expenditure ~ $15)

26. Herbal drugs in international health care • Economic aspects: Global market of herbal drugs, Nutraceuticals ~$60 billion with 6% annual growth rate. Major share of Chinese and Koreans. Indian share variously estimated at 0.35-3.0%. Chinese production increased 200% between 1995-1999. • Local acceptance: • Developed • USA: 42% use CAM spending over 29 billion US$ and 629 million visits in 1998. • UK: 28% use, spent 1.6 billion pounds and 127 million visits in 1998. • Australia: 60% use, A$ 620 billion in 1999. • Developing • Malaysia: Per capita consumption of traditional drugs, more than double of modern pharmaceuticals. • S.Korea: Per capita consumption of traditional drugs 36% more than modern drugs. • African countries: 9 to 10 patients attending hospital OPD have first consulted a traditional healer.

27. Advantages of herbal drugs • Modern drugs can produce serious side effects • Latrogenic diseases fourth leading cause of death in USA and other developed nations (JAMA, April 1998). • Side effects of drugs kill more Americans annually than the world war II and Vietnam war combined (M. Rath N. Y. Times 28.2.2003). • Around 2600 persons died in the Twin Tower tragedy on 11th September 2001 causing global repercussions. It is, however, not recognized that about the same number die in USA from side effects of prescription drugs every 10 days (JAMA, April 1998).

28. Herbal drugs are best suited for: • PRIMARY HEALTH CARE • INFECTIOUS DISEASES • AIDS and other viral infections • Opportunistic infections • MDR infections (e.g. T.B., Malaria) • DEGENERATIVE & GERONTOLOGICAL CONDITIONS • Osteoporosis • Chronic arthritis like osteoarthritis and rheumatoid arthritis • Neurological like Alzheimer, Parkinsonism • Anti-aging • Metabolic disorders • Diabetes • Dyslipidemias • Other conditions • Microcirculatory disorders • Liver diseases • Immunostimulants • Anti-cancer • Drugs affecting male libido

29. Standardization of Herbal drugsRaw Drugs • Passport data of Raw Plant Drugs (Crude drugs) • Correct taxonomic identification & authentication • Study on the medicinal part: root, stem, bark, leaves, flowers, fruits,nuts, gum, resins etc. • Collection details: Location, stage & development/ growth of the plants, time, pre-processing storage etc. • Organoleptic examination of raw drug: • Evaluation by means of sensory organs: touch, odour taste • Microscopic & molecular examination • Chemical composition (TLC, GLC, HPLC, DNA fingerprinting) • Biological activity of the whole plant • Shelf life of raw drugs

30. Standardization of Herbal drugs-Herbal Formulation • Follow defined Good Manufacturing Practices (GMP) • Scientific Verification • Toxicity evaluation • Chemical profiling • Pharmacodynamics – effect of drug in the body • Pharmacokinetics – absorption, distribution, metabolism, mechanism of action and execution • Dosage • Stability and shelf life • Presentation and Packing • Therapeutic merits – Compared with other drugs

31. Good Practices/Techniques in Herbal Products • Good Survey of literature (Ancient & Modern) • Develop and Observe Norms of: • Good Agricultural Practices (GAP) • Good Collection/Harvesting and Post Harvest Handling Practices (GCP/ GHP & GPHP) • Good Laboratory Practices (GLP) • Good Clinical Practices (GCP) • Good Manufacturing Practices (GMP) • Good Marketing Techniques (GMT)

32. Physico-chemical value Parameters required for quality evaluation of herbal drugs

33. Standardization & Quality Evaluation of Herbal drugs • Shape • External • Marking • Qualitative • Quantitative • SEM Studies • Powder Studies • Colour • Odour • Taste • Texture • Fracture Macroscopic Microscopic BOTANICAL ORGANOLEPTIC • Moist. Cont. • Extrac. Values • Ash Values • Fluores. Analy. QUALITY EVALUATION OF HERBAL DRUGS PHYSICAL Microbial Contamination BIOLOGICAL CHEMICAL • Toxicological • Pharmacological • Other specific activities HPTLC Finger printing Sec. Metabolites DNA Finger printing • Qualitative • Quantitative • Chromatography • Heavy metal • Pesticide residue • Mycotoxin Antagonistic HPTLC GLC HPLC • Bacterial • Fungal

34. ‘DARUHARIDRA’ - A Controversial drug of Ayurveda • Three plants used as ‘Daruharidra’ • In Northern India Berberis aristata DC. (Family- Berberidaceae), • In Southern part Coscinium fenestratum (Gaertn.) Colebr (Family- Menispermaceae), • In Easter part Coptis teeta Wall. (Family- Ranunculaceae) • Berberine is the active constituent present in all the three species thus on the basis of berberine all of these can be used as substitutes of ‘Daruharidra’. However, scientific investigation also suggest that different Berberis species viz. B. asiatica, B. chitra and B. lycium can also be used as substitute of ‘Daruharidra’ as all the species have almost similar chemical profile.

35. Different vernacular Names of Berberis spp. Arabic - Ambarbaris Bhutia - Tsema Bengali - Darvi Canarese - Bagisutra English - Indian Barbery, Tree turmeric Greek - Lykion indikon Garhwal Himalayas - Kingora, Kilmora, Kashmal Hindi - Daruhaldi, Darhald, Chitra Jaunsar - Kashmoi Japanese - Indo-ohbaku Latin - Berberis Malayalam - Maradarisina, Maramanjal Nepal - Chitra, Chotra Persian - Bedana, Chitra, Zirishk Punjab - Chachar Himanchal Pradesh - Kammul, Kashmal, Kaumul Sanskrit - Daruharidra, Darvi Tamil - Maramanjal, Mullukala Telugu - Kasturipaspu Urdu - Ambar 

36. Medicinal uses of Berberis spp. Ayurveda: It is bitter and at the same time hot and pungent useful in boils, diabetes, affections of ear, eye and mouth. The decoction is very useful preperation for opthalmic infection, cough, poison and boils. Local health traditions (folk remedies) of Berberis spp. in Northern India: B. aristata DC. and B. asiatica Roxb.: InGarhwal region (U. P.) : In Dhanulti and Raithal locality near to Uttarkashi of U.P. :Rasaut a root extracts is used for: Eye conjuctivitiis, Malarial fever, Skin diseases. In Kumaun region (U. P.) :  In Ranikhet locality of Nainital (U.P.) :Decoction of root bark is used in eye troubles. local people they use root extract for treating fever and skin diseases. Roots are also used for snake and scorpion bite. B. Chitria Lindl. InKangra valley (Himanchal Pradesh): The filtered decoction of root is used for treating conjunctivitis and other ophthalmic diseases. B. LyciumRoyle In Kangra vally(Himanchal Pradesh):The decoction of root is given orally as blood purifier. Raw fruits are also eaten for digestive disorders. Leaves are used as fodder. It is taken with the juice of radish for jaundice.

37. Market Survey of Berberis Spp. Roots of Berberis spp. are collected in fairly large quantities in Chamba District Of Himanchal Pradesh and in Tehri-Garhwal of Uttar Pradesh during Aug., Sept. and are being sold in the nearby markets or to the traders which finally reach the drug markets of India. Different market samples are identified as – Trichur samples - Stem of Coscinium fenestratum. Banglore samples - Root of Coscinium fenestratum. Amritsar, Aligarh, Dehradun, Hyderabad, Jammu, Lucknow, and Varanasi samples- the different Berberis species.

38. Major constituent of Berberis spp. Berberine

39. Quantitative Estimation of Berberine in different Berberis species

40. Quantitave estimation of berberine in different market samples of Berberis spp.

41. Comparative HPTLC profile of Berberine in different market samples

42. Densitometric scan of different samples of Berberis spp. at UV 266 nm

43. Berberis aristata D.C. Brief Taxonomic description B. aristataDC., Syst. Nat. 2:8.1821; Hook f. & Thomson in Fl. Brit. India 1:110. 1872 pp.: Naithani, Fl. Chamoli 1:46.1984; Sharad, LWG 221239, 1998. Berberis aristata DC. is a large deciduous shrub usually 1.8-3.6 m high; twigs whitish or pale yellowish brown, erect cylindrical, smooth and strongly striate; blaze 5-7.5 mm, bright yellow with coarse reticulate fibres; leaves 3.8-10 x 1.5-3.3 cm, obovate or elliptic, entire or spinous-toothed, base gradually narrowed, with prominent reticulate nerves, glossy dark green above and glossy pale green beneath; flowers numerous, stalked; inflorescence a simple drooping raceme, bracts small, linear, acuminate; sepals 8 or 9, imbricate, oval, petaloid, yellow; petals 6, in two whorls, strongly imbricate, concave, bright yellow veined with two oval linear glands at the base of the lateral veins; stamens 6 equal, hypogynous, opposite and slightly shorter than the petals; ovary simple, 1-celled, with a few erect ovules; style short, stigma peltate; fruit a small berry about 7-10 mm, ovoid or oblong ovoid, blue black with a whitish bloom tipped along with the persistent style and stigma. Flowering and Fruiting: August – October

44. Berberis aristata DC. – Macroscopic & Microscopic T.S. cellular structure of stem (x 100) Dried Root T.S. of the root (x 100) Fruits and Seeds TLS of the root (x100) Powder study (x400)

45. Botanical Analysis of root of Berberis aristata DC

46. Berberine HPTLC profile and densitometric chromatogram of B. aristataroot

47. THE AYURVEDIC THERAPEUTIC STRATEGY • Determine PRAKRUTI (Constitution) by -history taking -observations • NIDANA (Diagnosis) Nature, degree and extent of imbalance of Tridoshas. Library of 5800 clinical signs and symptoms in Ayurvedic texts • CHRONOBIOLOGY: Impact of season, time and environment on Tridoshas. • SWASTHAVRUTA: Life style modification • AHARA: Dietary modifications • PANCHAKARMA: Purification of the body • AUSHADHI: "Designer Medicine" unique for the particular patient prepared from a Pharmacopoeia utilising 1200 plants, 100 minerals and 100 animal products in numerous formulations.

48. OBJECTIVES OF RESEARCH ON AYURVEDIC DRUGS • Improved formulations and reduced number of Ayurvedic drugs • Use of GMP procedures and QC • Certified shelf life and improved dosage form • Validated indications and contraindications • Deletion of obsolete or toxic formulations • Use of Ayurvedic drugs in modern clinical practice • Inclusion in essential list of drugs • Adjunct to existing drugs • Treatment of diseases where modern drugs not available or unsatisfactory • Development of suitable formulations, standardized extracts or active constituents • IPR protection wherever feasible • Inclusion in Pharmacopoeias.

49. OBJECTIVES OF RESEARCH ON AYURVEDIC DRUGS • New indications for Ayurvedic drugs • Development of new drugs for Ayurvedic practice • Utilizing leads from other countries • Study of unscreened flora, specially endemic or threatened species • Studies on Ayurvedic drugs for veterinary use

50. Development of new drug/ novel uses for Ayurvedic drugs • Utilization of new leads on natural products from other countries. Several of these plants or related species are found in India • The vast unscreened flora of the country and published activity data on some of these • Study of further accessions from families yielding active plants