Biodiversity Wealth and Opportunities for Asia Pacific Countries in Biotechnology &

Herbal Technology

P. PushpangadanNational Botanical Research Institute

(Council of Scientific &Industrial Research),Rana Pratap Marg, Lucknow-226001

(September 2003)

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)

21st Century21st 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.

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

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)

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)

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’

Conservation of Biodiversity

Strategies & Priorities

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…….

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

Biodiversity & TK: Capital Assets of Asia-Pacific Countries

(Contd.)

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……..

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

Biodiversity & TK: Bridging the North- South Gap

Asia-Pacific Countries should:

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).

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.

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)).

• 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.

Relevant Provisions of TRIPs on Biological Resources

(Contd….)

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.

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.

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.

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.

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.

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.

Chemical Prospecting Drugs and pharmaceuticals Pesticides Cosmetics Food additives Other industrially valuable Chemical products

Bioprospecting

Gene Prospecting Genetic engineering Crop development Fermentation Cell culture

Bionic Prospecting DesignsSensor technologiesArchitecture Bioengineering Biomodeling

Bioprospecting: Linkages and leads

Biodiversity & IK/TK

Conservation Sustainable use

Benefit sharing

Bioinformatics

IPR

Biotechnology

Information technology

Herbal technology

Bioprospecting

Drug development

Pharmaceuticals

Agrochemistry

Cosmetics

Proteins

Enzymes

New crop varieties

GMOs

GM foods

Designs etc.

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.

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)

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.

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).

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 arthritisNeurological like Alzheimer, Parkinsonism Anti-aging

Metabolic disordersDiabetesDyslipidemias

Other conditionsMicrocirculatory disordersLiver diseasesImmunostimulantsAnti-cancerDrugs affecting male libido

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

Standardization of Herbal drugs-Herbal Formulation

Follow defined Good Manufacturing Practices (GMP) Scientific Verification

Toxicity evaluationChemical profilingPharmacodynamics – effect of drug in the bodyPharmacokinetics – absorption, distribution, metabolism, mechanism of action and executionDosageStability and shelf lifePresentation and PackingTherapeutic merits – Compared with other drugs

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)

Good Practices/Techniques in Herbal Products

Physico-chemical value

Parameters required for quality evaluation of herbal drugs

QUALITY EVALUATION QUALITY EVALUATION OF HERBAL DRUGSOF HERBAL DRUGS

CHEMICALCHEMICAL

BIOLOGICAL

BIOLOGICAL

ORGANOLEPTIC

ORGANOLEPTICBOTANICALBOTANICAL

PH

YS

ICA

LP

HY

SIC

AL

• Moist. Cont.• Extrac. Values• Ash Values• Fluores. Analy.

Macroscopic Microscopic

• Qualitative • Quantitative• SEM Studies • Powder Studies

•Shape

•External

•Marking

• Colour • Odour• Taste• Texture • Fracture

Antagonistic

Microbial Contamination

•Toxicological•Pharmacological

•Other specific activities

• Bacterial• Fungal

•Qualitative• Quantitative • Chromatography• Heavy metal • Pesticide residue• Mycotoxin

HPTLC GLC HPLC

HPTLC Finger printing Sec. Metabolites DNA Finger printing

Standardization & Quality Evaluation of Herbal drugs

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.

‘‘DARUDARU HARIDRA’HARIDRA’ - A Controversial drug of - A Controversial drug of AyurvedaAyurveda

Arabic - AmbarbarisBhutia - TsemaBengali - DarviCanarese - BagisutraEnglish - Indian Barbery, Tree turmericGreek - Lykion indikonGarhwal Himalayas - Kingora, Kilmora, KashmalHindi - Daruhaldi, Darhald, Chitra Jaunsar - KashmoiJapanese - Indo-ohbakuLatin - Berberis Malayalam - Maradarisina, MaramanjalNepal - Chitra, ChotraPersian - Bedana, Chitra, ZirishkPunjab - ChacharHimanchal Pradesh - Kammul, Kashmal, Kaumul Sanskrit - Daruharidra, DarviTamil - Maramanjal, MullukalaTelugu - KasturipaspuUrdu - Ambar 

Different vernacular Names 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. :

 In Garhwal 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.

 In Kangra valley (Himanchal Pradesh): The filtered decoction of root is used for treating conjunctivitis and other ophthalmic diseases.

 B. Lycium Royle

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.

Medicinal uses 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.

Market Survey of Berberis Spp.

N+

O

O

H3CO

OCH3

Major constituent of Berberis spp.

Berberine

3.55

1.417

2.25

1.024

5.2

1.4

3.99

2.005

0

1

2

3

4

5

6

RootStemRootStemRootStemRootStem

B. aristataB. asiaticaB. chitriaB. lycium

Berberine (%)

B. aristata RootB. aristata StemB. asiatica RootB. asiatica Stem

B. chitria RootB. chitria StemB. lycium RootB. lycium StemQuantitative Estimation of Berberine in different Berberis species

2.272

1.927

1.45

2.813.01

2.27

3.36

3.71

2.18

2.73

0

0.5

1

1.5

2

2.5

3

3.5

4

Locations

Berberine (%

)

AligarhAmritsarBangalore 1Bangalore 2DelhiJammuHyderabadLucknowTrichurVaranasi

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

Comparative HPTLC profile of Berberine in different market samples

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

B. aristata DC., 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

Berberis aristata D.C. Brief Taxonomic description

Berberis aristata DC. – Macroscopic & Microscopic

Dried Root

Fruits and Seeds

T.S. of the root (x 100)

T.S. cellular structure of stem (x 100)

TLS of the root (x100)

Powder study (x400)

Botanical Analysis of root of Berberis aristata DC

Macroscopic Woody, yellowish brown, cylindrical, knotty with thin brittle bark.Cut surface bright yellowFracture hard, texture short, odourless and bitter in taste.

Outline Circular

Cork cells Brown, 10-20 Layered, rectangular

Cortical zone

30-35 layered, parenchymatous, filled with tannin, starch grains and rhomboidal crystals of Ca-oxlate.

Sclereids Solitary or in group of 2 to 10

Pericyclic fibres

Mostly solitary but sometimes in groups of 2 to 10.

Alkaloidal contents

Present

Vessels Solitary or in group of 2 or 3

Medullary Rays

Heterogenous, 2 to 4 cells broad, pitted, filled with starch and alkaloidal content

HPTLC profile and densitometric chromatogram of B. aristata root

Berberine

THE AYURVEDIC THERAPEUTIC STRATEGY

1. Determine PRAKRUTI (Constitution) by -history taking -observations

2. NIDANA (Diagnosis) Nature, degree and extent of imbalance of Tridoshas. Library of 5800 clinical signs and symptoms in Ayurvedic texts

3. CHRONOBIOLOGY: Impact of season, time and environment on Tridoshas.

4. SWASTHAVRUTA: Life style modification5. AHARA: Dietary modifications 6. PANCHAKARMA: Purification of the body 7. AUSHADHI: "Designer Medicine" unique for the particular patient

prepared from a Pharmacopoeia utilising 1200 plants, 100 minerals and 100 animal products in numerous formulations.

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 formValidated indications and contraindicationsDeletion of obsolete or toxic formulations

Use of Ayurvedic drugs in modern clinical practiceInclusion in essential list of drugsAdjunct to existing drugs Treatment of diseases where modern drugs not available or unsatisfactory Development of suitable formulations, standardized extracts or active constituentsIPR protection wherever feasibleInclusion in Pharmacopoeias.

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

OBJECTIVES OF RESEARCH ON AYURVEDIC DRUGS

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

Vision of Herbal Drug Industry

“To provide intellectual capital to make available safe, cost effective, affordable therapeutics to the people of Indo-Pacific region – to help to reduce the percentage of mortality, morbidity and to emerge as significant players in the global market place.”

BIOPARTNERSHIP: RELEVANCE OF APCTT

Easy and regulated access to genetic resources & TK and biotechnologies

Exchange of information pertaining to conservation and sustainable use of biogenetic resources and associated TK

Mutually Agreed Terms

Prior Informed Consent

Equitable Benefit Sharing Agreement

APCTT can bring the Asia-Pacific Countries should come together to develop Strategies for:

Build up S&T CapabilitiesCapitalize biodiversity and TK for bioindustrial developmentInsulate from BiopiracyEnsure national sovereign rights over biodiversity and TK Empower local and indigenous communities, including womenBuild up location –specific biodiversity enterprises using local bio-resources and TK through S&T applications

BIOPARTNERSHIP: RELEVANCE OF APCTT

APCTT can help Asia Pacific Countries:

AREAS: Biodiversity & TK

Biotechnology

Bioinformatics

Bioprospecting

BIOPARTNERSHIP: RELEVANCE OF APCTT

APCTT can help in Capacity building and Training for Asia-Pacific countries

APCTT can help in in Capacity Building and Training for Asia-Pacific countries

STRATEGIES:

1. Development of Transparent Policies and Mechanisms to ensure:

Access to and transfer of genetic resources and technologies among participating countries

Evolving equitable benefit sharing models based on sustainable use and S & T based value addition to bioresources and associated TK

Contd...

STRATEGIES (Contd.):

2. Promotion of multi-country collaborative R&D projects on various facets of Bioprospecting, particularly herbal drug and pharmaceutical prospecting and other natural product development sectors

3. Generation of IPR- covered products, processes, technologies and services, and thereby converting the bio-resources and associated TK in to economic wealth of the country and its people.

Action ProgrammeImportant points that the biodiversity richthird world nations should undertake:

Complete inventory & documentation of all Biological resources including the microorganisms Check list/database of the floristic wealth of the nation along with the associated knowledge system Ground check to know the actual situation and identify the gaps: Study - genetic diversity, distribution pattern, association pattern and gradients Identify- rare, endemic and endangered status of spp. , if any.

Action Programme

Prepare -passport data of all important and endemic biodiversity. Passport data should cover morphological, cytological, chemical and molecular level (DNA/gene level) information so as to prevent bio/gene piracy.

Identification of problems and solutions in conservation, threatened status of species, ecosystems -with causes of threats.

Identification of problems and solutions in conservation.