Phytochemistry 56 (2001) 237243

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Phytochemistry and medicinal plants

J. David Phillipson *
Centre for Pharmacognosy, The School of Pharmacy, University of London, 29-39 Brunswick Square, London WC1N 1AX, UK Received 14 July 2000; received in revised form 1 August 2000

Abstract A truncated history of the contribution of plants to medicine is given with reference to some of the less well known ancestors of the Harborne family. Six of the top 20 prescriptions dispensed in 1996 were natural products and the clinical use of drugs such as artemisinin, etoposide and taxol has once more focussed attention on plants as sources of novel drug entities. High through-put robotic screens have been developed by industry and it is possible to carry out 50,000 tests per day in the search for compounds which have specicity of action against a key enzyme or a subset of receptors. Bioassay-guided fractionation of plant extracts linked to chromatographic separation techniques leads to the isolation of biologically active molecules whose chemical structures can readily be determined by modern spectroscopic methods. The role of academics in the search for new drugs is discussed by reference to some of our research into natural products with activity on the central nervous system, on pain receptors, the malaria parasite Plasmodium falciparum, the wound healing properties of the sap of species of Croton (Dragon's blood), and a traditional Chinese medicine used to treat eczema. Expertise in phytochemistry has been essential for this research and the strong lead shown by Professor Jerey Harborne is gratefully acknowledged. # 2001 Published by Elsevier Science Ltd.
Keywords: J.B. Harborne; Medicinal plants; Phytochemistry; Academics; New drugs; Central nervous system; Eczema; Malaria; Pain; Wound healing

1. Introduction The use of plants as medicines goes back to early man. Certainly the great civilisations of the ancient Chinese, Indians, and North Africans provided written evidence of man's ingenuity in utilising plants for the treatment of a wide variety of diseases. In ancient Greece, for example, scholars classied plants and gave descriptions of them thus aiding the identication process. Theophrastus has been described by some as the father of botany (Fig. 1) but little, if anything, has been recorded on his distant relative J.B. Theophrastus1 who extolled the virtues of medicinal plants and forecast the possibility of discovering avonoids. As Europe entered the dark ages much of this information would have been lost had it not been for the monasteries that acted as centres for the production of medicinal plants which were used to heal the suering of mankind. There is still much we can learn from investigating the old herbals,
* Tel.: +44-207-753-5800; fax +44-207-753-5909. E-mail address: profjdp@msn.com 1 The lecture presented made reference to imaginary forefathers of Jerey B. Harbone.

particularly those less well known such as the one attributed to the monk J.B. Harbonus1. It was not until the 19th century that man began to isolate the active principles of medicinal plants and one particular landmark was the discovery of quinine from Cinchona bark by the French scientists Caventou and Pelletier (Fig. 2). Much less is known about the isolation of quinine by J.B. Caventou1 and J.B. Pelletier1. Such discoveries led to an interest in plants from the New World and expeditions scoured the almost impenetrable jungles and forests in the quest for new medicines (Fig. 3). One of the lesser known intrepid explorers was J.B. van Harbon1 who was never happier than when he was able to hatchet his way through the jungle stripping o the barks from every tree in sight. Such expeditions would last for years and it was not until the plants arrived at a well equipped phytochemical laboratory that the real discoveries could be made (Fig. 4). Laboratories such as those of Professor J.B. de Harbonney1 became centres for the isolation of the active principles of medicinal plants from around the globe. Years of toil would be rewarded by the isolation of numerous avonoids which were welcomed by the cognoscenti as well as the rapidly expanding pharmaceutical companies.

0031-9422/01/$ - see front matter # 2001 Published by Elsevier Science Ltd. PII: S0031-9422(00)00456-8

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Fig. 1. Theophrastus father of botany.

Fig. 2. First of the alkaloid chemists; Caventou, Pelletier and Quinine.

2. New drugs from nature Prior to World War 2, a series of natural products isolated from higher plants became clinical agents and a number are still in use today. Quinine from Cinchona

bark, morphine and codeine from the latex of the opium poppy, digoxin from Digitalis leaves, atropine (derived from ()-hyoscyamine) and hyoscine from species of the Solanaceae continue to be in clinical use. The antibiotic era dawned during and after World War 2 due to

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Fig. 3. Wresting the Jungle's secrets.

Fig. 4. The development of chemotherapy.

the antibacterial eects of a whole series of natural products isolated from species of Penicillium, Cephalosporium, and Streptomyces. In the post-war years there were relatively few discoveries of new drugs from higher

plants with the notable exception of reserpine from the Rauwola species heralding the age of the tranquillisers and also vinblastine and vincristine from Catharanthus roseus which were eective in cancer chemotherapy.

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Despite these discoveries the impact of phytochemistry on new drug development waned and inevitably the innovative pharmaceutical industry turned to synthetic chemicals. Successful clinical agents emerged from multidisciplinary research teams in which pharmacologists and synthetic chemists collaborated, e.g. atenolol (beta-blocker) and captopril (ACE-inhibitor) for treatment of hypertension, salbutamol (adrenoceptor stimulant) for asthma and the benzodiazepines (hypnotics and anxiolytics) for insomnia and anxiety attacks. During recent years, the attention of the pharmaceutical industry has switched once more to the natural world and this may be illustrated by reference to three clinical drugs, taxol, etoposide and artemisinin (Phillipson, 1999a). Taxol is obtained from the bark of the Western Pacic Yew, Taxus brevifolia. The isolation and structure determination of taxol followed on from experiments that showed that a crude extract was active against cancer cells in laboratory tests. Although this activity was discovered in the early 1960's, it was not until 1971 that the structure elucidation of this complex diterpene was determined. In 1979 it was reported that the mode of action was through promotion of the assembly of tubulin into microtubules. Clinical trials did not take place until the early 1980's and it was not until the 1990's that taxol and its semisynthetic derivative taxotere were shown to be clinically eective against breast and ovarian cancers. The long period for the development of taxol as a clinical agent, its diculty in procurement as a natural product and the complexity of its chemical structure all attest to the diculties faced by the pharmaceutical industry in developing clinical agents from natural sources. The resin podophyllin obtained from the root of the mayapple, Podophyllum peltatum, is toxic and is used clinically to remove warts. The major constituent of the resin is the lignan podophyllotoxin which inhibits cell division. Because of its toxic properties it would seem to be not worthwhile pursuing any medicinal activities even though its eects on cell division would indicate potential use in cancer chemotherapy. However, a semisynthetic modied glucoside, etoposide, which has a dierent mode of action inhibiting topoisomerase II, has found clinical application in the treatment of lung and testicular cancers. Artemisinin is an unusual sesquiterpene endoperoxide that has been isolated as the active principle of the Chinese antimalarial herb Artemisia annua. Clinical trials have demonstrated that artemisinin is an eective antimalarial and can be used to treat infections of multidrug resistant strains of Plasmodium falciparum the cause of human malignant cerebral malaria. Semi-synthetic derivatives including artemether (the methyl ether of dihydroartemisinin) have improved pharmacokinetic properties and are also of current clinical use. The active moiety of artemisinin is 1,2,4-trioxane and a series of

synthetic analogues show remarkable activity against Plasmodium species in vitro and in vivo. Whether or not these will prove to be eective clinical agents or will lead to new clinical drugs is a matter for future research. The prospect of new drugs and medicines from plant sources is discussed further with reference to some of our research investigations (Phillipson, 1995, 1999a,b). 3. Will further new drugs be developed from natural product research? The clinical applications of taxol, etoposide and artemisinin have helped to revive an interest in higher plants as sources of new drugs (Phillipson, 1999a). Despite the belief that the majority of clinical drugs are synthetic in origin, it is interesting to note that 6 out of the top 20 pharmaceutical prescription drugs dispensed in 1996 were natural products and that over 50% of the top 20 drugs could be linked to natural product research. In recent years the development of sensitive biological testing systems, mainly by industry, has led to the procedure of high through-put screening. Such screens are carried out robotically and it is possible for a pharmaceutical or biotechnological company to run 50,000 biological tests per day. The test screens are based on specic enzymes within an animal or microbial biosynthetic pathway or on receptors or subsets of receptors. New screens are continually being introduced and batteries of compounds, synthetic and natural, are tested as screens come on line. Hence, banks of compounds or extracts are needed for industrial biological tests. It is estimated that there are some 250,000 species of higher plants and the majority of these have not been examined in detail for their pharmacological activities. Specic plants may have been subjected to particular tests, e.g. for cardiac activity, but they have not been examined for any other type of activity. The major screens for biological activities of plant extracts have been carried out in the search for new anticancer, antiviral and antifertility drugs. The development of the rapid screening tests now in use in industry has meant that many more plants can be evaluated for a wide range of biological activities. Unfortunately the results of such tests do not necessarily reach the public domain and are kept in locked industrial les. There still remains an urgent need to develop new clinical drugs and this can be exemplied by the numerous diseases which result from the malfunction of the central nervous system (CNS), e.g. Alzheimers and Parkinsons disease, epilepsy, migraine, pain, schizophrenia, sleeping disorders. Natural products already have a proven track record for CNS activities, e.g. caffeine, codeine, morphine, nicotine, reserpine and it is possible that there are further such drugs still to be found from nature (Phillipson, 1999b).

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With this in mind, we collaborated with two major international pharmaceutical companies, Glaxo (now GlaxoWellcome) and Pzer. In one investigation, some 10 Chinese plants were assessed for their activities against 18 radioligand-receptor binding assays which are implicated with CNS. The other investigation was concerned with pain and some 600 species of plant were tested. The pain receptors used were bradykinin II, neurokinin I, and a calcitonin gene related peptide. Half of the plants were selected from the ethnobotanical literature as being used for the treatment of pain and the other half were from a random sample. The results showed that there were more positive hits for activity in the biological screens for the selected group of plants (Phillipson, 1999b). 4. Malaria In 1996 it was reported that there were between 1.5 and 2.7 million deaths annually and that the majority of these were children. There are in the order of 500 million new incidences of malaria annually. It is without doubt one of the major threats to mankind and chemotherapy is hindered by the increase in drug resistant strains, particularly of Plasmodium falciparum. In the mid 1980's we posed the question `` Are new antimalarial drugs awaiting discovery from plants?'' Quinine, the rst eective antimalarial drug is still in clinical use and the more recently discovered artemisinin has proved to be an incentive for further research into plants. The only major scientic paper of antimalarial testing of plant extracts by the mid 1980's dated back to 1947 when it was reported that some 600 species of higher plant representing some 126 families had been tested against avian malarias. Several plants were active but the research pointed to two particular plant families, Simaroubaceae and Amaryllidaceae which had numerous active species. It is pertinent to ask why it took more than 30 years for this research to be followed through. The answer probably lies in the techniques which were available for carrying out this type of research. Avian malarias were used because they were the only tests for activity against Plasmodium species apart from those using monkeys. The avian tests which used live chickens and ducklings were notoriously dicult to carry out and were not thought to be necessarily predictive of activity against Plasmodium species which aected humans. These tests were not suitable for bioassay-guided fractionation of plant extracts. Furthermore, the chemical techniques available were also not suitable for this type of research. In the 1940's and 1950's there were not the sophisticated chromatographic separation techniques which are available today. Even if an active principle were isolated there were none of the spectroscopic techniques available for structure determination such as nuclear magnetic resonance spectro-

scopy, mass spectrometry or X-ray crystallography. By the mid 1980's not only were these chemical techniques available but also it was possible to test for activity against P. falciparum in vitro and there was a reliable test in mice against P. berghei (Phillipson, 1995). Following the lead from the 1947 paper, we tested activities of 5 species of Simaroubaceae against P. falciparum in vitro and utilised bioassay-guided fractionation techniques to isolate a series of active terpenoids (quassinoids). Some 40 quassinoids became available for structureactivity studies and this led to the preparation of semi-synthetic and synthetic analogues. Despite considerable research eort, no new clinical drug was developed from this work. Investigation of a range of plants used in traditional medicine for the treatment of malaria led to the isolation of a series of other compounds with activity against P. falciparum including isoquinoline and indole alkaloids, avonoids, mono-, di- and sesquiterpenoids. These results provided some scientic evidence which helped towards the justication of claims for the use of a number of traditional medicines and they also provided template molecules for synthetic approaches to new antimalarial drugs. Widening the range of biological tests to include other species of protozoa demonstrated activity of natural products against other tropical diseases which aect mankind, e.g. trypanosomiasis and leishmaniasis (Phillipson, 1995 and 1999a). 5. Do traditional medicines necessarily contain a single active ingredient? The isolation and use of natural products such as digoxin, morphine and quinine has resulted in replacing the plant extracts used with single chemical entities. There is a basic supposition that any plant possessing clinical eectiveness must contain an active principle which can completely replace the plant extract. Three examples from our research have shown that this may not necessarily be true (Phillipson, 1995). Artemisinin is without doubt the potent antimalarial active principle of Artemisia annua. Crude extracts of A. artemisia contain a plethora of other compounds including a series of avonoids and some of these enhance the activity of artemisinin against P. falciparum in vitro. Whether these ndings have clinical relevance has not been determined but they do lend support to the view that there may be some advantages to the medical use of extracts as opposed to isolated single entities. Dragon's blood is a term used for the blood red sap obtained from the bark of a number of S. American Croton species which are used for the treatment of wounds. The major constituents of the sap are polymeric anthocyanidins which co-occur with many minor constituents including diterpenes and simple phenols. Chemical and biological investigation of the properties

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of Dragon's blood led us to conclude that there is not one single wound healing principle. When the sap is used to cover a wound it forms a protective occlusive layer whilst some of the simple phenols act as potent antimicrobial agents and other compounds exert antiinammatory eects. In the 1980's it was noted by clinical dermatologists at Great Ormond Street Hospital for Sick Children in London that some of their young patients with severe atopic eczema were showing signs of improvement in their disease state. These improvements were not due to hospital therapy but to the co-administration of a traditional Chinese medicine (TCM). The patients had visited a TCM practitioner in central London and had been prescribed a multi-herbal prescription from which an aqueous extract was prepared for oral use. In 1992, it was reported that a double blind placebo controlled clinical trial of a ten herb mixture for oral use in children with non-exudative eczema conrmed substantial clinical benet as assessed by currently accepted Western orthodox medical practitioners. Our scientic investigations utilising an anti-inammatory/analgesic test with mice showed that four of the ten herbs possessed signicant activity in the mice but they proved to be inactive clinically in children. After some considerable investigation we concluded that not only was there no single active ingredient but also that it required all ten herbs to be present for clinical eectiveness. There are more than 12 dierent biological activities from the herbs in this TCM prescription including anti-inammatory, immuno-modulatory, anti-allergic, sedative and anti-pruritic. The chemical composition of the 10 herbs is a complex mixture of natural product molecules. 6. Conclusions Plants continue to be used world-wide for the treatment of disease and novel drug entities continue to be developed through research into their constituents. In the developed countries, high-throughput screening tests are used for bioassay-guided fractionation leading to the isolation of active principles that may be developed into clinical agents either as the natural product or a synthetic modication or a synthesised analogue with enhanced clinical action or reduced adverse side eects. Despite the massive arsenal of clinical agents developed by the pharmaceutical industry there has been an aversion by many members of the public and herbal remedies have proved to be popular as alternative or complementary treatments of disease. There is a need to evaluate herbal treatments by clinical trials using currently accepted protocols. In the developing countries large numbers of the World's population are unable to aord pharmaceutical drugs and they continue to use their own systems of indigenous medicine that are

mainly plant based. There is a great need to harness scientic and clinical research in order to investigate the quality, safety and ecacy of these herbal therapies. The aim of the pharmaceutical industry is to develop novel drug entities for the treatment of disease. Such drugs require specicity of action and are, for e.g. aimed at a particular subset of receptor. Although natural products continue to supply banks of compounds for new screens, the focus of industry is currently on combinatorial synthesis for new drug development. It must not be forgotten that natural products which result from millennia of biosynthetic pathways modied by evolution have a well established track record as medicinal agents and present a wide range of structural diversity. Drug development through natural product research is not without its problems and there is, for e.g. a need to eliminate common natural products such as saponins, tannins, etc. from plant extracts prior to testing by biological screening procedures. Academics can play a useful role in this area of research. They cannot match industry in the wide range of screens but they can use selective targets and collaborate with industry. This type of research needs a multi-disciplinary approach and this includes expertise in phytochemistry. It is a pleasure and an honour to present this lecture and to acknowledge the lead which Professor Jerey Harborne has given to Phytochemistry over so many years. I am one of those who owe a great debt to him and to the example which he has set. My own specialisation of Pharmacognosy was virtually wiped out from Pharmacy undergraduate curricula and for many years has been considered to be an outmoded area of research. Techniques in Phytochemistry have revolutionised our ability to investigate the medicinal agents present in plants and this is acknowledged by the industrial interest in plants over recent years. Thanks to the hard work and tenacity of Jerey Harborne we have been able to publish research articles in Phytochemistry and to continue working on the wealth of chemical diversity that exists in the plant kingdom. Acknowledgements I am grateful to Parke, Davis and Company for permission to reproduce the gures from the book Great Moments in Pharmacy by G.A. Bender, Detroit, Northwood Institute Press, 2nd Edition, 1967. References
Phillipson, J.D., 1995. A matter of some sensitivity. Phytochemistry 38, 13191343. Phillipson, J.D., 1999a. New drugs from nature it could be yew. Phytotherapy Research 13, 28. Phillipson, J.D., 1999b. Radioligand-receptor binding assays in the search for bioactive principles from plants. J. Pharm. Pharmacol. 51, 493503.

J.D. Phillipson / Phytochemistry 56 (2001) 237243 David Phillipson is Emeritus Professor of Pharmacognosy at the Centre for Pharmacognosy and Phytotherapy at The School of Pharmacy, The University of London. He was formerly Professor and Head of Department of Pharmacognosy at The School before retiring in 1994. In 1995, he was appointed for 6 months at The Chinese University of Hong Kong as Wilson T.S. Wang Distinguished International Visiting Professor. He is an Honorary Professor at the Chinese Academy of Medical Sciences, Insti-

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tute of Medicinal Plant Development, Beijing. For many years, he has been an active member of the Phytochemical Society of Europe and between 1977 to 1988 he held oces of Secretary, Vice-Chairman and Chairman. His research interests include the chemistry and biological activities of plants used in traditional medicine. He has received awards from the Phytochemical Society of Europe including the Tate and Lyle Award (1992), Medal (1994) and Pergamon Prize for creativity in plant biochemistry (1996). In 1989, he and four other European scientists in collaboration with Professor Meinhart Zenk (then of the University of Munich) were awarded the Korber Foundation Prize for achievement in European Science. The Pharmaceutical Society of Great Britain presented him with their Harrison Memorial medal in 1999.