15 Bioprospecting : Creating a Value for Biodiversity

Bioprospecting is the exploration of biological material for commercially valuable genetic and biochemical properties (Reid et al., 1993). This chapter will focus on the search for activities that could form the basis of new pharmaceuticals. Historically, most of the active ingredients in medicines have been natural products (Sneader, 1996), and natural products continue to form a productive source of new drugs (Newman and Cragg, 2007; Butler, 2008). Given that most drug discovery activity takes place in companies in the developed world and that most biodiversity is found in countries of the southern hemisphere, there needs to be a means whereby access to biodiversity is possible under terms and conditions that are mutually acceptable. After hundreds of years of unregulated collection of samples for many different purposes, the United Nations produced a framework for preserving the world’s biodiversity while encouraging the sustainable use of biodiversity. This Convention on Biological Diversity has been widely accepted, and it is discussed in the following section. The chapter will continue with descriptions of various attempts to calculate an economic value for biodiversity, followed by an outline of current bioprospecting practices.


Introduction
Bioprospecting is the exploration of biological material for commercially valuable genetic and biochemical properties (Reid et al., 1993). This chapter will focus on the search for activities that could form the basis of new pharmaceuticals. Historically, most of the active ingredients in medicines have been natural products (Sneader, 1996), and natural products continue to form a productive source of new drugs (Newman and Cragg, 2007;Butler, 2008). Given that most drug discovery activity takes place in companies in the developed world and that most biodiversity is found in countries of the southern hemisphere, there needs to be a means whereby access to biodiversity is possible under terms and conditions that are mutually acceptable. After hundreds of years of unregulated collection of samples for many different purposes, the United Nations produced a framework for preserving the world's biodiversity while encouraging the sustainable use of biodiversity. This Convention on Biological Diversity has been widely accepted, and it is discussed in the following section. The chapter will continue with descriptions of various attempts to calculate an economic value for biodiversity, followed by an outline of current bioprospecting practices.

United Nations Convention on Biological Diversity
The United Nations Convention on Biological Diversity (CBD) (www.biodiv.org) was one of the major outcomes of the Earth Summit in Rio de Janeiro in June 1992. The CBD has three main goals:  the conservation of biodiversity  the sustainable use of the components of biodiversity  the sharing of benefits arising from the commercial and other utilization of genetic resources in a fair and equitable way Signatories to the CBD recognise that countries have sovereign rights over their genetic and biological resources (i.e., biodiversity) within their boundaries, and agree to the conditions in the CBD for the preservation and sustainable use of biodiversity. In relation to accessing natural products for drug discovery, the CBD has a number of Articles (see Appendix) that set the tone for future interactions between companies and research organisations with countries with desired biodiversity. Biodiverse-rich countries that have ratified the CBD have to facilitate access to their biological resources (Article 15.2).
Nagoya Protocol is explicit about the responsibilities of signatories to create 'national focal points' and 'competent national authorities' to make available information on how to access genetic resources and traditional knowledge and to be responsible for granting access to biodiversity. In Africa, a diverse range of policies and laws relevant to access and benefitsharing is in place in some countries, but these are most developed in South Africa through the National Environmental Management: Biodiversity Act (10 of 2004) ('the Biodiversity Act') and the regulations passed under this Act in 2008. The Biodiversity Act requires bioprospectors to obtain a permit from the Government for bioprospecting involving indigenous biological resources, and for the export of these resources. Prior informed consent is required with landowners and indigenous communities before a permit is issued. Benefit-sharing agreements must be entered into with indigenous communities who use the resource traditionally, or who have knowledge of its properties (Wynberg et al., 2009).

Value of bioresources
A key issue in bioprospecting is benefit-sharing. However, much of the debate on this topic assumes that there are benefits to share. The historical successes in drug discovery based on natural products would suggest that there should be a continued appetite for accessing natural products for use in drug discovery programmes. Despite this, the pharmaceutical industry, in general, has reduced its use of natural products (Harvey, 2008), and there are few current examples of large-scale programmes designed to access a wide variety of natural products collected from their native environment. Moreover, advances in techniques for manipulating microbes to produce novels chemicals make the use of locally sourced bacterial samples more attractive to industry (Kingston, 2011). Never-the-less, it is reasonable to ask what an appropriate price might be for biodiversity, its conservation and its availability for bioprospecting. This is an area with much debate but little consensus (see, for example, Castree, 2003). Future royalty streams from blockbuster drugs (drugs that generate $1 billion in sales per year) directly or indirectly derived from bioresources were anticipated to contribute significantly to the conservation of biodiversity in source countries, and to the development of indigenous knowledge holder communities. However the age of blockbuster drugs seems to largely be over, and the pharmaceutical sector, as it is currently structured, is unable to deliver enough new products to market to generate revenues sufficient to sustain its own growth. Nearly all major drug developers are critically examining current R&D practices and, in some cases, considering a radical overhaul of their R&D models (Kaitlin, 2010). Only 13 natural productderived drugs were approved in the United States between 2005(Harvey, 2008, and FDA approvals of new drugs reached a 24-year low in 2007 (Li & Vederas, 2009). With the contraction and consolidation seen in the pharmaceutical industry, and with only 3 in 10 new products generating revenues equal to or greater than average pharmaceutical industry R&D costs (Kaitlin, 2010), it would seem that royalty streams from natural product-derived pharmaceuticals are too uncertain to be included in the valuation of biodiversity. A pragmatic approach to valuation is to focus on the monetary value paid by end users. In the early 1990s, Merck & Co. entered an agreement with the National Biodiversity Institute of Costa Rica, INBio, that provided a defined amount of cash (reported as $1 million) for a certain number of samples; although this deal is frequently mentioned in articles on the pros and cons of bioprospecting, key elements of the financial terms are still confidential, notably the payment per sample. More recently, GlaxoSmith Kline had an arrangement with the biotechnology company Extracta in Brazil: at its reported value of $3.2 million for a collection 326 of 30,000 samples (Dias & da Costa, 2007), this does not seem like a high price -about $100 per sample. The Swiss company Novartis also had an agreement with a Brazilian organisation, Bioamazonia, reported to be $4 million for 10,000 samples of micro-organisms. It should be noted that political concerns in Brazil about inappropriate commercialisation of its biodiversity caused both interactions to be terminated, highlighting the need for clear national focal points and competent authorities, as called for in the Nagoya Protocol. In a development from academic collaborations on natural products and drug discovery, the Strathclyde Institute for Drug Research, UK acted as a broker for natural product samples provided by its collaborators and offered samples under licence to companies for bioassay. Because each species could be used several times, the cumulative return on a single extract could be quite impressive, reaching $500-1,500 per gram. This can be compared with commodity prices obtained for a known herbal medicine that has proved active in doubleblind clinical trials: raw material costs for Hypericum perforatum (St John's wort) are $8-10 per kg. However, there is currently little demand from companies for samples of natural product extracts, and the brokerage activity has stopped. Another approach to valuation of biodiversity has argued that a value can be put on the potential contribution of an area of land to drug discovery (Simpson et al., 1996;Rausser & Small, 2000). Depending on the assumptions used, this varied from $21 per hectare (Simpson et al., 1996) to $9,177 per hectare (Rausser & Small, 2000). The latter valuation was thought to be sufficient to provide an economic reward to sustain biodiversity conservation. A later paper (Costello & Ward, 2003) sought to explain the very different valuations reached by the previous authors. They concluded that Rausser and Small's focus on species in biodiversity hotspots was largely responsible. Another approach to valuation of biodiversity used the pharmaceutical industry figures for costs and rewards of drug discovery and development to formulate a discounted cash flow model that gave a Net Present Value for an extract in a screening programme (Artuso, 1997). This was $487 per extract. A similar approach was used to explore the impact of varying the balance between upfront payments for sample supply and long-term royalties from sales of commercialised products (Lesser & Krattinger, 2007). In theory, more value is attached to a deal with an emphasis on royalty payments, but the authors note that larger up-front fees may be more of an incentive for conservation of biodiversity. The same approach was used to explore the potential added value of traditional knowledge and of pre-screened extracts. On average, use of samples suggested by traditional knowledge was expected to double the value of the deal. However, it is unlikely that this approach, which is based on averaging the theoretical outcome of testing large collections of samples, is appropriate for use of samples from traditional medicinal uses. These are more likely to be commercialised (if at all) on an individual basis. In addition to the possibility of providing a source for new chemical entities or derivatives, traditional medicinal plants may be commercialised as standardised plant extracts developed into polymolecular botanical medicines, dietary supplements or functional foods. The value of a registered botanical medicine may be considerable. In December 2005, the German Federal Institute for Drugs and Medical Devices (BfArM, Bonn) approved a new licence for the use of a proprietary extract of the root of the South African plant Pelargonium sidoides, (EPs® 7630), known as Umckaloabo, as a drug (Conrad et al., 2007). This registered liquid herbal medicine has been reported to have an annual turnover in Germany alone in 2006 of €80,000,000 (Brendler & van Wyk, 2008). In-country fractionation and screening could also, potentially, raise the value of a collection of natural product extracts on the basis of reducing the risk of failure to the commercial development partner. However, there has to be the necessary infrastructure and expertise to allow the screening, and the preliminary tests have to be relevant to the commercial partner (Lesser & Krattinger, 2007). The discussion above has only considered direct monetary benefits. However, both the Bonn Guidelines and the Nagoya Protocol list possible non-monetary benefits in addition to the monetary ones (Table 1). The non-monetary benefits need to be looked at seriously because technology transfer and improvements in capacity may contribute to sustainable development in biodiversity-rich countries, although that assumption has been questioned (Castree, 2003 Admittance to ex situ facilities of genetic resources and to databases; f.
Transfer to the provider of the genetic resources of knowledge and technology under fair and most favourable terms, including on concessional and preferential terms where agreed, in particular, knowledge and technology that make use of genetic resources, including biotechnology, or that are relevant to the conservation and sustainable utilization of biological diversity; g. Strengthening capacities for technology transfer; h. Institutional capacity-building; i.
Human and material resources to strengthen the capacities for the administration and enforcement of access regulations; j.
Training related to genetic resources with the full participation of countries providing genetic resources, and where possible, in such countries; k. Access to scientific information relevant to conservation and sustainable use of biological diversity, including biological inventories and taxonomic studies; l.
Contributions to the local economy; m. Research directed towards priority needs, such as health and food security, taking into account domestic uses of genetic resources in the Party providing genetic resources; n. Institutional and professional relationships that can arise from an access and benefit-sharing agreement and subsequent collaborative activities; o. Food and livelihood security benefits; p. Social recognition; q. Joint ownership of relevant intellectual property rights. Under the CBD, there is a clear need to reach agreement with the source of biodiversity on appropriate sharing of benefits arising from any commercialisation. There is also a commitment in the CBD to recognise and protect indigenous knowledge about uses of biodiversity. When it comes to benefit-sharing from commercial developments from such traditional knowledge, agreements can be hard to reach (see Boyd, 1996;Mays & Mazan, 1996). In part, this can be because of a cultural clash, e.g. where traditional knowledge is regarded as communal and not capable of being owned in a Western sense (see Cotton, 1997;Prathapan & Rajan, 2011). There can certainly be arguments over inventorship when the natural products in question are in widespread use, and there are frequent disputes about what constitutes appropriate benefits. In some cultures, monetary returns may have little meaning, and various attempts have been made to set up, for example, charitable foundations to distribute benefits in other ways (Mulholland & Wilman, 2003). Various access and benefit-sharing agreements have been analysed (Castree, 2003;Mulholland & Wilman, 2003;Medaglia, 2004;Laird & Wynberg, 2008). Clear criteria for success, in either economic or conservation terms, are missing so that it is difficult to reach objective conclusions. It seems to be extremely unlikely that bioprospecting will be a sufficient economic driver to support conservation of biodiversity. Indeed, the case of paclitaxel has been cited to show that a bioprospecting success can lead to negative consequences (Frisvold & Day-Rubenstein, 2008). Bioprospecting should perhaps be examined as an activity with some opportunities for local benefits and one that has to be regulated to make sure that it does not endanger biodiversity or deny fair and equitable benefits to indigenous knowledge-holders.

Historical successes in pharmaceutical bioprospecting
Bioprospecting (in terms of seeking leads for new drugs from natural products) can follow two main approaches: use of leads from traditional medical uses (i.e., from "ethnopharmacology"), and use of natural products as a highly diverse set of chemicals for random screening.

Traditional medicines
Historically, nature was the origin of all medicines (Sneader, 1996), and ethnopharmacology has provided some very notable past successes, including morphine (isolated in 1804), quinine (isolated in 1820), digitoxin (isolated in 1841), ephedrine (isolated in 1897), and tubocurarine (isolated in 1935). These compounds, or their analogues and derivatives, are still in widespread use. A further 50 examples are given by Cox (1994). More recent developments with an association with traditional uses include artemisinin and derivatives for malaria and prostratin as an anti-viral (see, e.g., Kingston, 2011). The development of a pharmaceutical product (as an appetite suppressant) from the traditionally used South African plants in the Hoodia genus was stopped, although the plant moved into development as a food supplement (Laird & Wynberg, 2008;van Heerden, 2008). Unfortunately, the early promise was not upheld and the commercial development rights have reverted to CSIR in South Africa. The approach to the commercial development of Hoodia was a case with inappropriate agreements between the South African research organisation CSIR and commercial development partners: the original agreements did not include the holders of the traditional knowledge, the San people. The absence of any benefits accruing to the San was subsequently successfully challenged by the South African San Council, and a mutually acceptable agreement was finally reached between the CSIR and the South African San Council after 18 months of negotiations (Wynberg et al., 2009). The more recent efforts to develop a medicinal or dietary supplement product from another plant (Sceletium tortuosum) originally used by the San may be instructive: the company involved, HG&H Pharmaceuticals (Pty) Ltd, successfully concluded a prior informed consent benefit-sharing agreement with the South African San Council and was also awarded the first bioprospecting and export permit to be issued by the South African Government (see http://www.zembrin.com/). There have also been leads from traditional medicines used as the starting point for the development of analogues that become the active ingredients of the final medicinal product. An example is podophyllotoxin, a compound isolated from Podophyllum peltatum, a plant used traditionally in North America for treating warts: this stimulated the work that led to the anti-cancer agent etoposide. Other examples can be found in the review by Newman and Cragg (2007), and more details and examples of ethnopharmacological investigations can be found in the books by Chadwick and Marsh (1994) and Cotton (1997). As discussed in the preceding section, accessing natural products used as traditional medicines can lead to many challenges -relating to ownership of intellectual property and benefit-sharing. It is also not necessarily a successful strategy for developing pharmaceutical products, as evidenced by the failure to date of either Shaman Pharmaceuticals or Phytopharm to commercialise new drugs or botanical medicines from traditional medicines.

Lucky finds
Random screening of natural products does not presuppose the existence of particular biological activities in any set of natural products: it relies on the assay to detect the activity.
The key to success is likely to be having the most chemically diverse collection of natural products, and this can be approached by using collections from diverse genetic sources. Notable successes include the development of cyclosporine A from a fungus (Tolypocladium inflatum) collected in Norway, and the development of rapamycin from a microbe (Streptomyces hygroscopicus) from Easter Island. The anti-cancer agent paclitaxel was discovered as a result of the National Cancer Institute's large-scale screening of plant extracts (Frisvold & Day-Rubenstein, 2008). Conservationists have highlighted the fact that 70% of the world's plant species and more than 60% of the world's vertebrate species are found on 1.4% of the land area of the world, and that some of the regions containing the greatest biodiversity are being threatened by development (Mittermeir et al., 1999). The richest regions have been defined as 25 mega diverse "hotspots" on the basis of the number of species found there and the high proportion of endemic species, i.e. those that occur naturally only in that region. The hotspots are shown in Table 2 along with the number of plant species they contain. Collections from areas with high endemism would be expected to yield many unusual compounds. However, apart from INBio's efforts in Costa Rica, there have been no systematic and widespread collections of samples from the biodiversity hotspots for bioprospecting. Seventy percent of the earth's surface is covered by sea, and the marine environment contains examples of most types of organisms. There may be more than 10 million species of marine macro fauna (Poore and Wilson, 1993) and many more species of marine microorganisms. There are few marine-based collections for bioprospecting: possibly, those of Magellan BioScience (www.magellanbioscience.com) in USA and MarBank in Norway. There has been relatively little work on bioprospecting such marine biodiversity, although salinosporamide from a marine bacteria is in early-stage clinical trials and ecteinascidin 743 (trabectedin) from a marine tunicate has been approved in Europe for the treatment of some cancers.
New and unusual biodiversity is still being discovered as unusual habitats are being sampled (Harvey 2007). Since this biodiversity has never been available for biological testing, it can be predicted that novel chemicals with potential as drug leads will be discovered if such biodiversity can be accessed for screening. However, this is largely driven by small-scale academic endeavours. Few companies are currently involved in providing access to biodiversity samples. Table 3 lists companies cited in 2001 as being active in this area (Harvey, 2002). Apart from Albany Molecular Research (AMRI), all have gone out of business or changed strategy. Analyticon still provides screening samples, but these are prepared as synthetic modifications of natural product scaffolds rather than purified natural products.

Current practices in bioprospecting
Most bioprospecting is currently performed on a small scale by numerous academic groups throughout the world. There are some larger programmes based on multi-group collaborations. These include the various International Cooperative Biodiversity Groups (ICBG) funded by the NIH in the USA and efforts coordinated by individual universities such as Rutgers in New Jersey and Strathclyde in Scotland. ICBG programmes involve US institutions and commercial companies with overseas participants in Costa Rica, Fiji, Indonesia, Madagascar, Panama, Papua New Guinea, the Philippines, and Vietnam and Laos (Cao & Kingston, 2009;Kingston, 2011). Rutgers University hosts a relatively new initiative: the Global Institute for BioExploration, GIBEX. This is an international network that aims to promote successful drug discovery from biodiversity through developing pharmacological screening methods that can be readily transferred to groups in partner countries (see http://www.gibex.org/). The University of Strathclyde has a long history of research in phytochemistry in collaboration with research groups in institutes overseas. This formed the basis for the creation of a worldwide network for drug discovery based on natural products. A highly diverse collection of plant extracts was assembled (covering more than 90% of the world's plant families) and used in drug discovery screening assays, either with a commercial partner or through collaborations between members of the network (see http://www.sidr.org/). Very few large pharmaceutical companies have maintained a strong presence in natural product-based drug discovery. Novartis has developed extensive collaborations with a few academic centres in the Far East, notably in China and Thailand. AstraZeneca had a long connection with a group based at Griffith University in Queensland. This continues within the Eskitis Institute (see http://www.griffith.edu.au/science-aviation/eskitis-institute-cellmolecular-therapies). There are smaller specialist companies involved in bioprospecting. These include MerLion Pharmaceuticals in Singapore (http://www.merlionpharma.com/) and Sequoia Sciences in St Louis, Missouri (http://www.sequoiasciences.com/). A different approach has been developed to make use of the structural diversity of isolated natural products: in silico drug discovery or virtual screening. In this, the chemical structures and physico-chemical properties of compounds are gathered in a computerised database that can be searched to find matches either to complement the three-dimensional structure of a drug target or the chemical features of a compound with the desired activity. This has been used at the University of Strathclyde in its Drug Discovery Portal (www.ddp.strath.ac.uk).
Chemists can submit structures to the Portal's database and biologists can propose therapeutic targets. Compounds are screened in silico by computational chemists within the Portal against the targets and the relevant chemists and biologists are notified of any predicted hits. The chemists then supply compounds to the biologists for testing on real assays against the target. Because chemists can only enter compounds into the Portal's database if they can guarantee that they can supply the compounds and because biologists can only suggest targets if they have relevant assays available, the reduction to practice can be very rapid after the initial in silico screening. The unique chemical database is rapidly expanding (currently over 14,500 compounds), with academic scientists from 21 institutions in five continents contributing, and the compounds have been shown to be highly diverse but still generally drug-like in their properties Harvey et al., 2010;Schuster & Wolber, 2010). Another approach to using natural products and virtual screening has been developed at the University of Innsbruck, Austria. Several databases of natural products have been created: a large general natural product database ("NPD") including over 110,000 compounds of molecular weights between 150 and 700, a small database of about 10,000 constituents from medicinal plants mentioned by Dioscorides ("DIOS" database) and a TCM database of about 10,000 compounds known from plants used in Traditional Chinese Medicine (Rollinger et al., 2004;). The assumed advantage of using compounds from known medicinal plants is that they may be less likely to be toxic than randomly sampled constituents.

Future prospects
Bioprospecting has been proposed as a potential means to encourage the conservation and sustainable use of biodiversity. The legal framework under the auspices of the United Nations is slowly being implemented by biodiversity-rich countries, but much still needs to be done if there is to be a genuine facilitation of bioprospecting. Perhaps the implementation of the Nagoya Protocol will provide the necessary impetus. However, the appetite for bioprospecting by pharmaceutical development companies has clearly diminished since the Rio Earth Summit in 1992, partly because of the complexities relating to access and benefit-sharing, often in the absence of adequate national regulatory clarity and institutional capacity. Despite the continuing appearance of successful drug development projects based on natural products, there is a sentiment that this approach may be too old-fashioned to be considered seriously: screening of natural products for new leads. Various technical problems undoubtedly exist with the screening and isolation of natural products, but the rewards for overcoming them would seem to justify the effort required, and technical solutions are being described in the literature. For example, purification and identification of natural products are believed to be difficult and slow: high throughput separation methods coupled with sensitive analytical techniques can resolve this (Bugni et al, 2008;Hu et al, 2008). Natural products are chemically complex: comparisons of the chemical properties of collections of natural products show that they more closely match the "chemical space" of successful drugs than collections of synthetic chemicals (Grabowski and Schneider 2007;Ganesan 2008). Natural products are reputed to give too many false positives on modern screening assays, but phenotypic assays are becoming more and more popular and it has been suggested that natural products, with their drug-like properties, are well-matched to such cell-based approaches, and extracts of natural products can be processed to remove reactive compounds or even convert them into novel drug-like structures (Rishton 2008). Natural products may only be available in small amounts: techniques for direct synthesis (Sunazuka et al, 2008) or production by molecular biology (Kennedy 2008) have been rapidly developing. While there is certainly no single "best" way to conduct drug discovery, just as there is not a single panacea for all ailments, it is surely time for a fresh look at the relatively unexplored opportunities provided by modern approaches to applying natural products in drug discovery. Perhaps the lead will have to be provided by the numerous academic groups active in bioprospecting. However, these groups would stand more chance of success if they could pool resources and work towards finding validated lead compounds that are likely to be suitable for development into medicines for unmet therapeutic needs. The growth of translational research and the establishment of centres of translational research will enable academic groups to become essential partners in pharmaceutical innovation. appropriate, and in accordance with Articles 16 and 19 and, where necessary, through the financial mechanism established by Articles 20 and 21 with the aim of sharing in a fair and equitable way the results of research and development and the benefits arising from the commercial and other utilization of genetic resources with the Contracting Party providing such resources. Such sharing shall be upon mutually agreed terms. Article 16. Access to and Transfer of Technology 1. Each Contracting Party, recognizing that technology includes biotechnology, and that both access to and transfer of technology among Contracting Parties are essential elements for the attainment of the objectives of this Convention, undertakes subject to the provisions of this Article to provide and/or facilitate access for and transfer to other Contracting Parties of technologies that are relevant to the conservation and sustainable use of biological diversity or make use of genetic resources and do not cause significant damage to the environment. 2. Access to and transfer of technology referred to in paragraph 1 above to developing countries shall be provided and/or facilitated under fair and most favourable terms, including on concessional and preferential terms where mutually agreed, and, where necessary, in accordance with the financial mechanism established by Articles 20 and 21. In the case of technology subject to patents and other intellectual property rights, such access and transfer shall be provided on terms which recognize and are consistent with the adequate and effective protection of intellectual property rights. The application of this paragraph shall be consistent with paragraphs 3, 4 and 5 below. 3. Each Contracting Party shall take legislative, administrative or policy measures, as appropriate, with the aim that Contracting Parties, in particular those that are developing countries, which provide genetic resources are provided access to and transfer of technology which makes use of those resources, on mutually agreed terms, including technology protected by patents and other intellectual property rights, where necessary, through the provisions of Articles 20 and 21 and in accordance with international law and consistent with paragraphs 4 and 5 below. 4. Each Contracting Party shall take legislative, administrative or policy measures, as appropriate, with the aim that the private sector facilitates access to, joint development and transfer of technology referred to in paragraph 1 above for the benefit of both governmental institutions and the private sector of developing countries and in this regard shall abide by the obligations included in paragraphs 1, 2 and 3 above. 5. The Contracting Parties, recognizing that patents and other intellectual property rights may have an influence on the implementation of this Convention, shall cooperate in this regard subject to national legislation and international law in order to ensure that such rights are supportive of and do not run counter to its objectives.