Fundamental Aspects of Bio Prospecting

Fundamental Aspects of Bio Prospecting – By Dilip Kumar Mandal and Samit Ray

Introduction:

Bio prospecting (biodiversity prospecting) involves searching for, collecting and deriving genetic material from samples of biodiversity that can be used in commercialized pharmaceutical, agricultural, industrial or chemical processing end products.

In other words, it is the exploration of microorganisms, wild plants and animals for commercially valuable genetic and bio-chemical resources.

In many cases, bio prospecting is a search for useful organic compounds in microorganisms, plants and fungi that grow in extreme environments such as rainforests, deserts and hot springs.

Most of the raw materials for biotechnology oriented industry comes from the wilds dwindling rainforests of the southern hemisphere. For example, many scientists believe that the cures for AIDS, cancer and other diseases lie hidden beneath these green vegetation.

So, bio prospecting involves finding the precious gene or cell line first and then patent the invention, since genetic materials from the developing world may yield cure for diseases as well as yield cash benefits through market. Agencies, scientific and corporate, use the folk wisdom of indigenous people to locate and understand the use of medicinal plants.

Then this knowledge is commercially exploited. Humankind has been studying, manipulating and exploiting natural diversity ever since the emergence of Homo sapiens over 150,000 years ago. Our early ancestors explored biodiversity and learned how to derive benefits from nature.

Early bio prospecting led to the improvement of methods for growing food, building shelters and maintaining health. Modern-day bio prospecting is simply an extension of our long history of exploring nature to improve the quality of our life.

The main objectives of bio prospecting are to fulfill economic and conservation goals and to augment medical and agricultural advances needed to combat disease and sustain a growing human population. However, business enterprises should form partnership with academician and with indigenous people if it wants to avoid paying high prices for the ‘green gold’ (the plants and animals with properties which businesses could use for new products and services) of bio prospecting.

Academic Work and Bio Prospecting- Case of Taq Polymerase:

Dr. Thomas Brock (1966) studied microorganisms living in yellow stone’s hot springs. He named one of the curious microorganism he isolated Thermus aquaticus. This microorganism lives and thrives in water so hot that it would kill an ordinary animal. Dr. Brock grew Thermus aquaticus in the laboratory and gave a living sample to the American Type Culture Collection for safe keeping. Dr. Brock’s work was an academic one.

In 1985, a biotechnology company named Cetus Corporation was developing a new way to duplicate genetic material. Dr. Kary Mullis, working at Cetus corporation invented a way to duplicate DNA, called Polymerase Chain Reaction (PCR).

But the high temperatures required by PCR destroyed the polymerase enzymes and fresh enzymes had to be added throughout the PCR process. Scientists at Cetus isolated an enzyme, named Taq polymerase, from Thermus aquaticus, which can withstand high temperatures of PCR process. PCR using Taq polymerase was so effective that a whole new scientific field has flourished as scientists finally had a convenient way to study DNA. Dr. Brock’s academic work in Yellow stone had a practical application that he never imagined in his academic career.

Biotechnology and Bio Prospecting:

Bio prospecting is often erroneously confused with biotechnology. Bio prospecting does frequently use advanced biotechnological processes and techniques. But it is only a very small subset of the vast field of biotechnology and two must be treated separately. Bio prospecting use biotechnological techniques to search for desirable characteristics in genetic material.

Non-genetic biological resources that involves indigenous knowledge of ethno botany is also a subject of bio prospecting. For example, bio prospecting may use recombinant gene technology to store target genes in retrievable form in representative environmental gene collection.

A bio prospecting firm does this by cloning or splicing DNA fragments into a vector. The DNA fragment spliced into the vector DNA is called a recombinant molecule or clone. A representative gene collection is the collection of these clones representing the entire DNA isolated from the organisms in a specimen.

In an attempt to derive an enzyme that allows the paper-making process to use less environmentally harmful bleach such efforts may be useful. Target compounds can now be reproduced in a laboratory by genetic engineering.

When a gene is identified in a wild organism that controls production of a useful compound, the compound-producing gene is inserted into laboratory bacteria and as the bacteria grow, they produce more of the compound which humans can then use. Pharmaceutical manufacturers use such biotechnology techniques to produce insulin from microbial stocks.

Bio Prospecting and Indigenous Peoples:

Indigenous traditional knowledge help to easily identify valuable chemical compounds from plants, animals and microorganisms. These knowledge are collected from territories traditionally inhabited by indigenous people. Between 1956 and 1976 the U.S. National Cancer Institute screened over 35,000 plants and animals for anti-cancer compound.

The programme was terminated in 1981 because of its failure to identify a greater number of new anti-cancer agents. A retrospective study concluded that the success rate in finding valuable species could have been doubled if medicinal folk knowledge had been used to target species. Scientists have found that 86% of the plants used by Samoan healers displayed significant biological activity in the laboratory tests. Crude extracts of plants used by one healer in Belize, yielded four times as many positive results in laboratory tests for anti-HIV activity than did specimens collected randomly.

It is generally accepted that one in 10,000 chemicals derived from mass screening of plants, animals and microbes eventually results in a potentially profitable drug. By contrast, Shaman Pharmaceuticals (now Shaman Botanicals), the U.S. based bio prospecting firm, that collects plants by talking to indigenous healers and watching them work, claims a success rate of 50%.

About half of the plants collected by Shaman’s researchers come up positive in screening tests, and it makes the indigenous knowledge 5,000 times more effective than random collection. This approach is less expensive and more focused. A knowledge about the working of indigenous remedies and preparation offer clues to the nature of chemical compounds involved.

In India, a well known example of bio prospecting and benefit sharing is the Kani- TBGRI (Tropical Botanic Garden & Research Institute) – model in Kerala. Kani is a tribal community inhabiting the southern Western Ghats in Kerala. In 1987, scientists from the TBGRI undertook an ethno botanical study in the tribal inhabited Western Ghat regions in Kerala.

They got an interesting ethno botanical information on a wild plant Trichophus zeylanicus, locally called “Arogyapacha” by the kani tribe. The Kani tribals accompanying the scientists, reported that eating fruits of this plant kept them energetic and agile. The Kani men were initially reluctant to reveal the source of fruit. The scientists convinced the Kani men that information would be used to conduct scientific investigation and if any marketable products developed, the benefits accrued would be shared with the tribe. Then the Kani men showed the plant, identified as Trichophus zeylanicus.

Later, pharmacological studies of the fruit confirmed its anti-fatigue properties. The leaves of the plant contained various glycolipids and other non-steroidal compounds with anti-stress and anti-hepatotoxic properties. The scientists developed a polyherbal formulation by Arogyapacha Ayurvedic pharmaceutical methods which was named ‘Jeevni’. After clinical evaluation this herbal drug was released for commercial production.

The manufacturing license of ‘Jeevni’ was transferred to the Aryavaidya Pharmacy, Coimbatore Ltd. for a license fee of Rs. 10 lakhs for a period of 7 years. TBGRI has agreed to share 50% of license fee and royalties with the tribal community. In November 1997, a number of Kanis with the assistance from TBGRI, registered a trust called Kerala Kani Samudaya Kshema Trust, comprising nine members from the tribals.

The Objectives of the Trust Deed Include:

(a) welfare and development activities for the Kanis of Kerala;

(b) preparation of biodiversity register to document the Kanis’ knowledge base,

(c) evolving and supporting methods to promote the sustainable use and conservation of biological resources.

Pharmaceutical Bio Prospecting and Phytomedicine:

There should be clear understanding of the difference between pharmaceutical bio prospecting and herbal medicine or phytomedicine. Pharmaceutical bio prospecting generally requires minimal sample quantities. It involves a high return and a low-volume harvesting is required as it strives to synthesize compounds from a limited amount of natural products. Phytomedicines involve high volume harvesting. High-volume harvesting may endanger the biological diversity in general and threaten a particular species. Thus pharmaceutical bio prospecting does not ordinarily endanger biodiversity but current practices in phytomedicine industry always pose a threat to biodiversity.

On the other hand, phytomedicine industry harvests various parts of plants and animals for consumption with minimal processing because of the high costs involved for a relatively low-return industry. In other words, the danger to biodiversity lies more in current practices of phytomedicine industry than in pharmaceutical bio prospecting. Conservation practices should be applied to phytomedicine industry. For this, forest farms can be organized to grow and harvest targeted plants for commercial use. Medicinal plants can be cultivated as follow crops.

The Role of Pharmaceutical Industry in Bio Prospecting:

A growing number of pharmaceutical corporations, biotechnological companies are stalking the forests, fields and waters of the developing world in search of biological riches and indigenous knowledge. Northern-based institutions seek access to tropical biodiversity for the primary purpose of developing patented and profitable products. Conservation and equity remain secondary issues.

The medicinal plants and microbes from the South contribute at least $30 billion a year to the North’s pharmaceutical industry. One conservative estimate states that the market for natural product research specimens within the U.S. pharmaceutical industry alone is $30-60 million per annum.

Today, there is greatly renewed interest in natural product screening for medicinal compounds. In 1980, none of the U.S. pharmaceutical industry research budget was spent on research on higher plants. Now, it is estimated that over 200 companies and research organizations worldwide are screening plant and animal compounds for medicinal properties.

In mid-1980s, one industrial analyst warned that, at current extinction rates of plants and animals, the Earth is losing one major drug every two years. Each medicinal plant lost in the tropical rainforests could lose drug firms possible sales of more than $200 million. Only less than 1% of the world’s 250,000 tropical plants have been screened for potential pharmaceutical applications. Around 80% of people in developing countries rely on medicines largely based on plants and animals.

With the advances in molecular biology and the availability of more sophisticated tools for screening, it is increasingly cost effective for pharmaceutical industries to conduct natural product research. In a high-technology regime, extracts from biological specimens undergo rapid and precise screening procedures that allow for the isolation of active compounds displaying a specifically targeted activity. As a result the market for exotic biological specimens is expanding rapidly.

Bio Prospecting for Conservation and Sustainable Development:

Two Fundamental Issues Concerning Bio Prospecting are:

(a) Sustainable use of biological resources and their conservation and

(b) The economic development of source countries and local communities.

The search for useful products derived from biological resources coupled with innovative ways to link benefits with conservation of biodiversity and sustainable economic development is attracting much attention worldwide. For thousands of years, biodiversity has been the source of useful compounds and materials for food, energy, shelter, medicines and environmental services. The pharma­ceutical industry has benefited from biodiversity through drug development from natural compound, while the agricultural industry improves crops by breeding them with wild relatives.

About half of the current best selling pharmaceuticals are from natural products. The combined market world wide for pharmaceuticals, agrochemicals and seeds is over $400 billion per year and the genetic diversity provides the starting material for a major portion of this market.

The issue of conservation is closely related to pharmaceutical industry. The loss of natural resources in the form of species extinctions accompanied by habitat loss is important to pharmaceutical companies in their search for novel natural product. Amongst the species being lost will be some which have the ability to produce important, but as yet undiscovered, chemical molecules.

In case of plant species, the issue of collection and re-collection are to be considered. In the event of a successful discovery, it is imperative that raw materials be available in sufficient quantities for follow-up work and / or compound production at a reasonable scale.

A balance must be reached between small initial collections requiring further follow-up and maintaining availability of samples. Larger initial collections may restrict the diversity of samples available for collection. For example, recollection issue is extremely important with marine algae. Marine algal species are difficult to grow under artificial conditions.

This means that the natural source of algal materials must be judiciously conserved and sustainably managed to ensure continuing supply. Pharmaceutical industry’s involvement in conserving biodiversity in situ is essential due to variations in secondary metabolite production between different samples of the same species from different habitats.

It is not clear, however, whether these differences mirror the diversity of genotypes within a sampling site, or if the diversity is a result of habitat variability or epigenetic variability. Studies undertaken with marine algae to examine this have highlighted the complex relationship that exists between genotype, environment and phenotype as expressed in secondary metabolic product.

Recently, bio prospecting has acquired increased attention as the developing countries seek to conserve their biodiversity and also share benefits from bio prospecting. Bio prospecting offers a potentially interesting approach to link biodiversity conservation with the biotechnology sector.

But its success will depend upon the ability of these efforts in isolating useful compounds at a cost comparable to other techniques of drug development. It is too early to say the extent of success to be achieved with bio prospecting in the area of conservation and economic development.

It is considered that from the application of genetic engineering in agricultural research, the world will get most of its food, fuel, fibre, chemical, feedstock, and even some pharmaceuticals from genetically modified plants. The mega diversity countries have significant stake for harnessing the potential of bio prospecting for sustainable economic development.

Compensated Bio Prospecting:

By the early 1990s, objections to uncompensated bio prospecting that does not share benefits with the source country became contentious. Since 1992, the Convention on Biological Diversity (CBD) has embodied the principles of compensated bio prospecting globally.

Prior to that, no formal international guidelines existed to compensate host countries when bio prospecting by outsiders eventually led to commercial products. Host countries received nothing even as they witnessed outsiders reap profits from native biodiversity. The following example would suffice to illustrate the condition.

In 1969, a scientist from Switzerland, working for Sandoz laboratories, took a holiday in Norway and collected soil samples from the Hardangervidda mountain plateau in south central Norway. These soil samples were subjected to Sandoz’ test procedure in Switzerland. In 1972, a strong immunosuppressive was found in cyclosporine, produced by the fungus Tolypocladium inflatum.

In 1983, the U.S. Food and Drug Administration approved the use of cyclosporine to prevent organ transplant rejection. In 1997 alone, Novertis (new name of Sandoz, after merger with Ciba-Geigy in 1996) earned $1.2 billion from this drug. Norway was never compensated. In a situation of regulated access regime, Norway could have had a reasonable claim of 2% of annual royalties from Novartis’s sale of cyclosporine medicines. And this would have amounted to $24 million for 1997.

The CBD recognizes that states have sovereign rights over their natural resources and that terms and conditions for access to these materials are within the domain of national legislation.

The Convention also recognizes the ‘knowledge, innovations and practices of indigenous and local communities” and specifically “encourage the equitable sharing of benefits arising from the utilization of such knowledge, innovations and practices”. The convention sanctions bilateral agreements by making repeated reference to “mutually agreed terms” for access to genetic materials, subject to prior informed consent” of the state.

Compensated bio prospecting involves obtaining prior informed consent from the source country, sharing benefits and promoting sustainable use of biodiversity. Where indigenous knowledge holders are involved, efforts made to recognize and protect their rights. Benefits can take various forms, from royalties to negotiated advance and milestone payments, capacity building, facilities and equipment transfer, personnel training, sharing of research and other forms.

Negotiated Bio Prospecting Agreements:

Currently, a negotiated agreement is the preferred tool used to provide compensation for access to biodiversity. Specific terms of compensation are left to the parties to agree upon incorporating the principles embodied in the Conventions on Biological Diversity. Negotiated agreements that provide balanced rights and benefits for its parties are preferred to the more problematic working out of intellectual property rights.

The first major bilateral contract for bioprospecting was made public in September, 1991 (prior to CBD) when Merck & Co. (a U.S. pharmaceutical corporation) announced a 2-year, $1.35 million deal with the Instituto Nacional de Biodivesidad (INBio) of Costa Rica, a private, non-government research institute.

Under the contract, INBio will allow Merck drug screening from wild plants, insects and microorganisms. In return, Merck will provide INBio a 2-years research budget of $1.35 million, an undisclosed share of royalties on any resulting commercial products and technical assistance and training to help establish drug research in Costa Rica. InBio also agreed to contribute 10% of its up-front payment from Merck and 50% of any royalties it may eventually receive to Costa Rica’s National Park Fund to help conserve nation parks.

Although the Merck / INBio agreement was hailed as a model agreement for bioprospecting, it ignores the rights and roles of indigenous people. Since the announcement of the Merck / INBio agreement, other contracts between Northern-based corporations / institutions and southern-based research institutes / government agencies have followed.

Convention on Biological Diversity (CBD):

The role of CBD in bio prospecting and sharing of natural resources has already been mentioned in earlier part of this article. The CBD was opened for signature on June 5, 1992 at the United Nations Conference on environment and development in Rio de Janeiro in Brazil. The CBD is an international agreement that applies to sovereign nations.

This international protocol gave expression to the novel idea that the bioprospecting process could yield conservation and development benefits. Carried out properly, the process could provide economic incentives for the conservation of biodiversity as well as for local and regional development.

It should also be noted that the CBD is widely considered an unforceable protocol that relies on voluntary compliance among the parties. Article 1 of the CBD states its objective as: The conservation of biological diversity, the sustainable use of its components and the fair and equitable sharing of the benefits arising out of the utilization of genetic resources including by appropriate access to genetic resources and by appropriate transfer of relevant technologies taking into account all rights over those resources and to technologies and by appropriate funding.

Regardless of the CBD and its principles, various environmental groups and researchers continue to view bio prospecting as initiative of the North to globalize the control, management and biological diversity of resources which lie primarily in the third world.

Bio Prospecting and Bio Piracy:

Bio piracy refers to the appropriation, generally by means of patents, of indigenous biomedical knowledge by foreign entities without compensatory payment. Bio piracy is also used to cover the various forms of power in balance between richer and poorer countries which arise out of poorer countries tendencies towards high biodiversity and richer countries’ wanting the benefits of that high biodiversity. Bio piracy is a value-laden term which has established itself as the primary concept in academic ethical debate on the subject.

For political correctness, the combination “bio piracy and bio prospecting” can be used. Bio piracy and bio prospecting can be easily defined in terms of each other – bio piracy is illegal or unethical bio prospecting, bio prospecting is legalized or ethical bio piracy.

The classic case of bio piracy is the Rosy Periwinkle (Madagaskar Periwinkle), a plant native to Madagascar. Research into the plant was prompted by the plants’ traditional medicinal role and resulted in the discovery of a large number of biologically active chemicals, including the children’s cancer cure vincristine. Vincristine was initially patented and marketed by Eli Lilly and earned millions of dollars annually but the country of origin did not receive any payment.

This particular case is a good example to show that bio piracy cases are not as simple as they seem.

Complicating Factors are:

(a) The Rosy Periwinkle, while, native to Madagascar, had been widely introduced into other tropical countries around the world before the discovery of vincristine. This meant that researchers could obtain local knowledge from one country and plant samples from another,

(b) The locally known medical properties of the plant were not the same as the medical properties discovered and commercially used by Eli Lilly. The use of the plant as a cure for diabetes was the original stimulus for research, but cures for cancer were the most important result,

(c) Different countries are reported to have acquired different believes about the medical properties of the plant.

In 1995 the U.S. Department of Agriculture and a pharmaceutical firm received a patent on a technique to extract an antifungal agent from the neem tree (Azadirachta indica), which grows throughout India. Indian villages have long understood the tree’s medicinal values. As a result of widespread public outcry, legal action by Indian government followed, with the patent eventually being overturned in 2005.

Future Road-Map of Bio Prospecting:

1. Characterization of biodiversity in each and every climatic zones.

2. Bio prospecting of novel genes, biomolecules and enzymes.

3. Crop improvement including proteins, fats and carbohydrates.

4. Bio prospecting deep sea bed – Antioxidant, antifungal, anti-HIV, antibiotic, anti­cancer, anti-tuberculosis and anti-malaria drugs are derived from ocean organisms. Applications for the treatment of Alzheimer’s disease, cystic fibrosis are also under consideration. Deep-sea ecosystem hold the promise of huge potential to future human well-being.

5. Bio prospecting for novel organisms for the production of bio-diesel and bioremediation.

6. Investigating bacteriophages as a source of new treatment for drug-resistant bacteria.

7. To conserve the biodiversity.

8. To protect indigenous knowledge that will ensure our learning of nature’s benefits.