Context: Whether natural product drug discovery programs should rely on wild plants collected "randomly" from the natural environment, or whether they should also include plants collected on the basis of use in traditional medicine remains an open question.Objective: This study analyzes whether plants with ethnomedical uses from Vietnam and Laos have a higher hit rate in bioassay testing than plants collected from a national park in Vietnam with the goal of maximizing taxonomic diversity ("random" collection).Materials and Methods: All plants were extracted and subjected to bioassay in the same laboratories. Results of assays of plant collections and plant parts (samples) were scored as active or inactive based on whether any extracts had a positive result in a bioassay. Contingency tables were analyzed using χ2 statistics.Results: Random collections had a higher hit rate than ethnomedical collections, but for samples, ethnomedical plants were more likely to be active. Ethnomedical collections and samples had higher hit rates for tuberculosis, while samples, but not collections, had a higher hit rate for malaria. Little evidence was found to support an advantage for ethnomedical plants in HIV, chemoprevention and cancer bioassays. Plants whose ethnomedical uses directly correlated to a bioassay did not have a significantly higher hit rate than random plants.Discussion: Plants with ethnomedical uses generally had a higher rate of activity in some drug discovery bioassays, but the assays did not directly confirm specific uses.Conclusions: Ethnomedical uses may contribute to a higher rate of activity in drug discovery screening.

The ICBG (International Cooperative Biodiversity Groups) program, through which institutions located in biotechnology-rich countries in the North collaborate with institutions located in the biodiversity-rich countries in the South (with the support of an industrial partner) to discover and develop natural-product drugs, is an experiment in the design of bioprospecting efforts. This chapter describes the general aims and organization of the ICBGs and describes in great detail the agreements that governed the University of Illinois at Chicago-Vietnam-Laos ICBG. The chapter includes material concerning IP (intellectual property) rights issues, informed consent, various forms of benefit sharing (including the sharing of shortand long-term, namely, royalty, benefits), capacity building, and community reciprocity. It offers a model for other such agreements. HANDBOOK OF BEST PRACTICES | 1511 One model of a biodiversity prospecting effort is a program called ICBG (International Cooperative Biodiversity Groups). Based in the United States, ICBG falls under the auspices of the Fogarty International Center (FIC) of the United States National Institutes of Health (NIH). It also collaborates with the National Science Foundation (NSF) and the U.S. Department of Agriculture (USDA).3 A five-year cycle program, it went into operation in 1993 in response to a request for applications issued by FIC in 1992.4 The ICBG second cycle began on 1 October, 1998, as a result of new and recompeting proposals in response to a request for applications issued by FIC in 1997.5 On 17 October 2002, a request for applications for a 2003–2008 ICBG cycle recompetition was again issued.6 2. The ICBG pRoGRAm International Cooperative Biodiversity Groups, or ICBGs, address the interdependent issues of drug discovery, biodiversity conservation, and sustainable economic growth. They are founded in the belief that efforts to examine the medicinal CHAPTER 16.5

The impact of the University of Illinois at Chicago-based Vietnam-Laos International Cooperative Biodiversity Group (ICBG) Program "Studies on Biodiversity of Vietnam and Laos", which has been in operation for the period of 1998-2005, touches on five major areas of endeavor: (a) biodiversity inventory and conservation; (b) studies on medicinal plants; (c) drug discovery and development; (d) economic development; and (e) issues on intellectual property rights and benefit sharing in natural products drug discovery and development. Highlights are presented and the significance is discussed.

Ethnobotany/ethnopharmacology has contributed to the discovery of many important plant-derived drugs. Field explorations to seek and document indigenous/traditional medical knowledge (IMK/TMK), and/or the biodiversity with which the IMK/TMK is attached, and its conversion into a commercialized product is known as bioprospecting or biodiversity prospecting. When performed in a large-scale operation, the effort is referred to as mass bioprospecting. Experiences from the mass bioprospecting efforts undertaken by the United States National Cancer Institute, the National Cooperative Drug Discovery Groups (NCDDG) and the International Cooperative Biodiversity Groups (ICBG) programs demonstrate that mass bioprospecting is a complex process, involving expertise from diverse areas of human endeavors, but central to it is the Memorandum of Agreement (MOA) that recognizes issues on genetic access, prior informed consent, intellectual property and the sharing of benefits that may arise as a result of the effort. Future mass bioprospecting endeavors must take heed of the lessons learned from past and present experiences in the planning for a successful mass bioprospecting venture.

The Convention on Biodiversity mandates a new approach to the discovery of natural product drugs, one that incorporates concepts of national ownership of genetic resources, intellectual property rights in traditional knowledge, and sharing of economic benefits with countries that are the source of new natural products. The International Cooperative Biodiversity Group (ICBG) program was established to support experimentation in implementation of the Convention through development and execution of international agreements for bioprospecting. The agreement of one such ICBG program, between the University of Illinois at Chicago and institutions in Vietnam and Laos, is presented here. The core elements contained in the single, five-way Memorandum of Agreement are the arrangements for intellectual property rights, treatment of informed consent, and plans for benefit-sharing (including the sharing of short- and long-term royalty benefits, capacity building, and community reciprocity). Program participants were able to develop a practical and flexible agreement that satisfies the wishes of all institutions that are parties to it.

An International Cooperative Biodiversity Group (ICBG) program based at the University of Illinois at Chicago initiated its activities in 1998, with the following specific objectives: (a) inventory and conservation of of plants of Cuc Phuong National Park in Vietnam and of medicinal plants of Laos; (b) drug discovery (and development) based on plants of Vietnam and Laos; and (c) economic development of communities participating in the ICBG project both in Vietnam and Laos. Member-institutions and an industrial partner of this ICBG are bound by a Memorandum of Agreement that recognizes property and intellectual property rights, prior informed consent for access to genetic resources and to indigenous knowledge, the sharing of benefits that may arise from the drug discovery effort, and the provision of short-term and long-term benefits to host country institutions and communities. The drug discovery effort is targeted to the search for agents for therapies against malaria (antimalarial assay of plant extracts, using Plasmodium falciparum clones), AIDS (anti-HIV-l activity using HOG.R5 reporter cell line (through transactivation of the green fluorescent protein/GFP gene), cancer (screening of plant extracts in 6 human tumor cell lines - KB, Col-2, LU-l, LNCaP, HUVEC, hTert-RPEl), tuberculosis (screening of extracts in the microplate Alamar Blue assay against Mycobacterium tuberculosis all performed at UIC, and CNS-related diseases (with special focus on Alzheimer's disease, pain and rheumatoid arthritis, and asthma), peformed at Glaxo Smith Kline (UK). Source plants were selected based on two approaches: biodiversity-based (plants of Cuc Phuong National Park) and ethnobotany-based (medicinal plants of Cuc Phuong National Park in Vietnam and medicinal plants of Laos). At mc, as of July, 2001, active leads had been identified in the anti-HIV, anticancer, antimalarial, and anti- TB assay, after the screening of more than 800 extracts. At least 25 biologically active compounds have been isolated, 13 of which are new with anti-HIV activity, and 3 also new with antimalarial activity. At GSK of 21 plant samples with a history of use to treat CNS-related diseases tested to date, a number showed activity against one or more of the CNS assay targets used, but no new compounds have been isolated. The results of the drug discovery effort to date indicate that tropical plant diversity of Vietnam and Laos unquestionably harbors biologically active chemical entities, which, through further research, may eventually yield candidates for drug development. Although the substantial monetary benefit of the drug discovery process (royalties) is a long way off, the UIC ICBG program provides direct and real-term benefits to host country institutions and communities.

Bioassay-directed fractionation of an extract prepared from the dried leaves and stem barks of Ficus fistulosa Reinw. ex Blume (Moraceae) led to the isolation of verrucarin L acetate (1), together with 3alpha-hydroxyisohop-22(29)-en-24-oic acid, 3beta-gluco-sitosterol, 3,4-dihydro-6,7-dimethoxyisocarbostyril, 3,4,5-trimethoxybenzyl alcohol, alpha-methyl-3,4,5-trimethoxybenzyl alcohol, indole-3-carboxaldehyde, palmanine, and aurantiamide acetate. Roridin E (2) was identified in a subfraction from the dried leaves and stems of Rhaphidophora decursiva Schott (Araceae). Verrucarin L acetate and roridin E were characterized as macrocyclic trichothecene sesquiterpenoids and found to inhibit the growth of Plasmodium falciparum with IC 50 values below 1 ng/ml.

Bioassay-directed fractionation of an extract prepared from the dried leaves and stem barks of Ficus fistulosa Reinw. ex Blume (Moraceae) led to the isolation of verrucarin L acetate (1), together with 3alpha-hydroxyisohop-22(29)-en-24-oic acid, 3beta-gluco-sitosterol, 3,4-dihydro-6,7-dimethoxyisocarbostyril, 3,4,5-trimethoxybenzyl alcohol, alpha-methyl-3,4,5-trimethoxybenzyl alcohol, indole-3-carboxaldehyde, palmanine, and aurantiamide acetate. Roridin E (2) was identified in a subfraction from the dried leaves and stems of Rhaphidophora decursiva Schott (Araceae). Verrucarin L acetate and roridin E were characterized as macrocyclic trichothecene sesquiterpenoids and found to inhibit the growth of Plasmodium falciparum with IC 50 values below 1 ng/ml.

Bioassay-directed fractionation led to the isolation of 14 compounds, six of which possess antimalarial activity, from the dried leaves and stems of Rhaphidophora decursiva. Polysyphorin (1) and rhaphidecurperoxin (6) showed strong activities against Plasmodium falciparum. Rhaphidecursinol A (2), rhaphidecursinol B (3), grandisin (4), and epigrandisin (5) were less active against the same organism. Among the isolates, rhaphidecursinol A (2) and rhaphidecursinol B (3) were determined to be new neolignans, and rhaphidecurperoxin (6) is a new benzoperoxide. Known compounds isolated include polysyphorin (1), grandisin (4), epigrandisin (5), (+)-medioresinol, (-)-pinoresinol, (-)-syringaresinol, (+)-glaberide I, (+)-dehydrovomifoliol, (-)-liliolide, (-)-hydroxydihydrobovolide, and N-butylbenzamide, of which compound 1 appears worthy of further evaluation as an antimalarial agent. Structure elucidation and identification were accomplished by spectroscopic means including 1D and 2D NMR analyses.

The UIC-based ICBG is a multidisciplinary, international research program, comprised of one academic institution (PCRPS/UIC, the base institution), two research institutions (NCST, Vietnam: RIMP, Laos). one national park (CPNP, Vietnam), and one industrial partner (GW). The goals of this ICBG are (a) to perform inventory and to promote the conservation of biodiversity of Vietnam and Laos; (b) to search for biologically active compounds front plants of Vietnam and Laos as potential candidates for drug development: and (c) to promote economic development in host countries (development of human resources, including strengthening of institutional infrastructure, and improvement of standard of living of communities participating in the project). The project's efforts are targeted to terrestrial organisms, specifically, the seed plants (angiosperms and gymnosperms) in Vietnam and Laos, while the therapeutic focus includes antimalarial, anticancer, antiviral/anti-AIDS therapies, and therapy against diseases of the central nervous system, including pain and Alzheimer's disease. For the implementation of the project, the 5-member Group is bound by a 5-way Memorandum of Agreement that defines the obligations and responsibilities of each member, and the joint obligations and responsibilities of all members together: Implementation of the project is effected through individual bilateral subcontractual agreements between the UIC and other member institutions. Members agree that more than 51% of royalty stream may accrue will return to the countries providing genetic material that led to the discovery and commercialization of a compound.

From Ficus fistulosa two triterpenoids 3β-acetyl ursa-14:15-en-16-one and lanosterol-11-one acetate were isolated along with five known triterpenoids, 3β-acetyl-22,23,24,25,26,27-hexanordamaran-20-one, 24-methylenecycloartenol, sorghumol (isoarborinol), 11α,12α-oxidotaraxeryl acetate, and ursa-9(11):12-dien-3β-ol acetate. Their structures were elucidated on the basis of spectral data interpretation.

Nitrate reductase deficient (NR-) cell lines isolated on N. tabacum (1.3) and N. plumbaginifolia (4) fall into two phenotypic classes: Nia- mutants, which are defective in the NR apoprotein, and Cnx- mutants, which are defective in molybdenum cofactor essential for NR and xanthine dehydrogenase. Protoplast fusion has been used to study allelism of these mutants. Previously, two complementation groups (nia, cnxA) were determined in N. tabacum (2, 6) and four groups in N. plumbaginifolia (4, 5). The objective of the present work is to test newly isolated Cnx- mutants of N. tabacum (3) and to see if complementation of NR occurs in interspecific hybrids.

Summary Protoplasts of Arabidopsis thaliana race Columbia were prepared from fast growing suspension culture cells. About 50–60% of the protoplasts formed calli when plated at a density of about 3 × 105 protoplasts per ml in the medium of Gamsorg et al. (1968) modified to contain 1 mg 2,4-D, 0.15 mg BA and 1000 mg CaCl2 × 2H2O per liter and 0.4 M glucose. About 40% of these calli formed shoots when transferred onto plant regeneration medium.

Protoplasts of Arabidopsis thaliana race Columbia were prepared from fast growing suspension culture cells. About 50–60% of the protoplasts formed calli when plated at a density of about 3 × 105 protoplasts per ml in the medium of Gamsorg et al. (1968) modified to contain 1 mg 2,4-D, 0.15 mg BA and 1000 mg CaCl2 × 2H2O per liter and 0.4 M glucose. About 40% of these calli formed shoots when transferred onto plant regeneration medium.