Ayurvedic Herbal Drugs in the Treatment of Malignant Swellings

[Full title: Selection and Evaluation of Ayurvedic Herbal Drugs which might be Useful in the Treatment of Malignant Swellings / By Hobbe Friso Smit, Herman Johan Woerdenbag, Jan Hinderikus Zwaving, Ram Har Singh, Rudi Paul Labadie]

Although the origin of Ayurveda goes back to a far past, the practice of medicine in India has developed ever since. The aim of this article is not to describe Ayurvedic topics as they come to us through the vast corpus of classical literature, but to select herbal drugs as a lead for anti-tumour activity by means of careful observation and interpretation of the traditional system of medicine in Northern India, as it is understood today. For the sake of clarity, a limited view on Ayurveda has been chosen, which served our purpose. Other approaches are of course not excluded. Introduction In Ayurveda, all matter is thought to be composed of five mahabhutas, namely akasa, vayu, tejas or agni, jala, and prthivi. The main constituents of the human body are dosas, dhatus and malas, which are derivatives of these pancamahabhutas. Dosas are physiological factors of the body and are categorised into vata, pitta and kapha. Vata is associated with movement, pitta with biotransformation and kapha with cohesion. These three dosas determine the prakrti, the constitution of an individual. The dhatus are seven in number: rasa, rakta, mamsa, medas, asthi, majjan and sukra. The three main malas are urine, faeces and sweat. For the burning and the transformation of the food in the body, there are thirteen agnis. One, the jatharagni, is responsible for the digestion of nutritious substances. Digestion takes place in three stages: madhurapaka, amlapaka, and katupaka. Five agnis, the pancabhutagnis, are responsible for the processing of the pancamahabhutas, each agni for a corresponding bhuta. Seven dhatvagnis are responsible for the transformation of each dhatu into the next one of the series. From the absorbed nutritious substance (ahararasa), rasadhatu is produced first. From rasa, rakta is formed, then mamsa, medas, asthi, majjan and sukra are produced successively (Sharma, P.V. 1976; Dash 1971, 66-77; 81-90; Singhal/Tripathi/Chaturvedi 1981, 233; Sharma, P.V. 1981-1985, vol. 2; Pandey 1987, 5-39; Dash 1989, 32-37). Under normal conditions, the dosas, dhatus and malas correspond to certain standards with regard to their quantities, qualities and functions. However, this situation is not static, and due to several endogenous and exogenous facWe have dispensed with also giving the common synonyms of the Sanskrit expressions cited above.

mahabhuta prthivi, jala Figure 1 The 20 gunas with their mahabhuta composition according to Sharma, P.V. 1976, p. 46 nya ning bung bung mahabhuta tejas/agni, vayu, akasa jala sita usna tejas/agni 114 Journal of the European Ayurvedic Society 5 (1997) tors, the dosas may become unbalanced, resulting in disease. Every disease is related to an imbalance of the dosas (Srikantha Murthy 1987, 16 f.; 42-44). There are said to be six cognisable entities in Ayurveda (padarthas), namely: dravya, guna, karman, samanya, visesa and samavaya. All dravyas are composed of the pancamahabhutas and are cognised through guna and karman, which are inherent in dravya. Guna is the property of a drug with regard to the body. There are forty-one gunas, but usually twenty are used, grouped in ten pairs, one guna being opposite another (fig. 1). Apart from these jala snigdha ruksna prthivi, tejas/agni, vayu prthivi, jala manda tiksna tejas/agni prthivi sthira sara vayu jala, akasa mrdu || kathina |prthivi prthivi, tejas/agni, vayu, akasa visada picchila jala tejas/agni slaksna khara vayu tejas/agni, vayu, akasa suksma sthula prthivi prthivi sandra drava jala

H.F. Smit, H.J. Woerdenbag, et al., Ayurvedic Herbal Drugs 115 gunas, there are other properties of dravya, which are: rasa, vipaka, virya, and prabhava. Rasa is the taste of a drug. There are six rasas: madhura, lavana, amla, katu, tikta and kasaya. Each rasa is composed of two of the five mahabhutas. Vipaka, the state of the ingested material after digestion, is generally madhura, amla or katu. The virya of a drug is one of the factors responsible for its action, and is expressed in terms of the strongest gunas, e.g. usna virya. Prabhava is the specific property which can not be explained by guna, rasa, vipaka and virya. Karman is the action of a drug on the body and can be samsodhana or samsamana. Karman can also be expressed in terms of the three dosas (Sharma, P.V. 1976, 3-67; Singhal/Chunekar 1982, 175-208). Arbuda² In the classical sources of Ayurveda, equivalents to the modern denomination 'cancer' are not found. However, some diseases are described which might be associated with tumours. One of these diseases is arbuda. MonierWilliams 1990 translates this as 'swelling, tumour, polypus'. It describes the clinical state of a disease which is associated with local swelling. Although several diseases meet this clinical description (e.g. inflammatory diseases and warts), neoplasms might be among them. An arbuda is said to have a big size and a round shape, to be immovable and deep-seated, and to grow slowly. The swelling is experienced as slightly painful and seldom suppurating (Ca,Ci 12,87; Su,Ni 11,13-15 a;21). Several types of arbuda are mentioned in the Ayurvedic literature. Usually they are classified according to their origin, e.g.: vataja, pittaja and kaphaja arbuda from vitiated dosas; raktaja, mamsaja and medoja arbuda from vitiated dhatus (Su,Ni 11,14 b). Furthermore, arbudas which arise in specific organs are mentioned, e.g.: netrarbuda: eye (Su,Ut 3,24), karnarbuda: ear (Su,Ut 20,5;16 b,c), nasarbuda: nose (Ca,Ci 26,109 a; Su,Ut 22,19 a), talvarbuda: palate (Su,Ni 16,44) and lingarbuda: penis (Su,Ni 14,14 a). Another disease which might be associated partly with tumours is gulma (Ca,Ci 5), which is translated by Monier-Williams 1990 as 'a chronic enlargement of the spleen or any glandular enlargement in the abdomen'. Other diseases associated with swelling are: sotha (Ca,Su 18), plihodara (Ca,Ci 13,37), yakrddalyudara (Su,Ni 7,16), kacchapa (Su,Ni 16,43 a), kanthasaluka (Su,Ni 16,51), gilayu (Su,Ni 16,58), sataghni (Su,Ni 16,57), pratyasthila (Su,Ni 1,91), vatasthila (Su,Ni 1,90), karnini (Su,Ut 38,15 b) and kakanaka kustha 2 References to Ca, Su and Ah are taken from Sharma, P.V. 1981-1985, Singhal 1981-1982, and Srikantha Murthy 1991-1992, respectively.

(Ca,Ci 7,20) (Bajracharya 1987). Pathogenesis In Ayurveda the course of every disease is described in a general model. Such a model is based on the changes in the dosa-balance and the subsequent consequences. The process of pathogenesis of arbuda can be described likewise. Except for the dosas, the metabolic processes as such also play an important role (fig.2). By means of several causative factors, for example, heavy and unctuous food, kapha can be aggravated (Su,Su 21,23). This affects the jatharagni and causes mandagni (Ah,Sa 3,73;76). In the case of mandagni the food is not digested properly and ama is formed by the mixing of aggravated dosas. This affects the dhatus in such a way that changes in their qualities occur, and they become vitiated. Another explanation is that, because of mandagni, the first dhatu, rasa, is not formed properly, becomes vitiated and accumulates. This is known as ama (Ah,Su 13,25-27). During the circulation of vitiated rasa and vitiated dosas through the body, all kinds of pathological processes can take place. The body-channels can become obstructed (srotorodha) (Ca,Ci 15,37). If a srotas is completely blocked, the rasa and the vitiated dosas might follow an alternative route (vimargagamana). Furthermore, the rasa and the vitiated dosas will get obstructed through abnormalities of the srotas of the dhatus and will not be able to reach the dhatus (Su,Su 24,10). All these pathological factors, i.e. vitiated dosas, vitiated dhatus, malfunctioning of agnis, formation of ama, will lead to a disturbed transformation of the dhatus, especially in the rakta, mamsa and the medas (Su,Ni 11,13 a). Because the mamsa nourishes the skin (Ca,Ci 15,17), a pathogenic defect of the muscular tissue might result in the injury of the rohini, the sixth layer of the skin. If this process continues, it might result in the formation of an arbuda (Su,Sa 4,4). Adhyarbuda and dvirarbuda can also occur (Su, Ni 11,20). Eventually, this can lead to the death of the patient (Dwarakanatha 1986; Bajracharya 1987, 8-10). Material and Methods The objective of this research project was to find out which Ayurvedic plants can be used as a lead for anti-tumour activity. To achieve this, the general model for the pathogenesis of arbuda, described above, was used to form criteria for the selection of plants from a list of herbal drugs which are used in Ayurveda. This list was prepared from literature. In this study the

H.F. Smit, H.J. Woerdenbag, et al., Ayurvedic Herbal Drugs 117 nidana (causative factors) ↓ sancaya accumulation of kapha →mandagni ↓ ↓ formation of ama ↓ ↓ ↓ formation of samarasadhatu ↓ prakopa excitation of kapha prasara spreading of excited dosa → srotodusti -> ↓ ↓ ↓ srotorodha ↓ vimargagamana ↓ sthanasamsraya localisation of excited dosa -> ↓ ↓ ↓ dosadusyasammurchana ↓ vitiation of mamsa ↓ vitiation of rohini ↓ ↓ purvarupa ↓ vyakti manifestation of disease -> rupa i.e. arbuda ↓ bheda complications Figure 2 Model of pathogenesis of arbuda according to Ayurveda

generally accepted identity of the traditional plant names mentioned in literature was used (Singh/Chunekar 1972; Vaidya 1982). This identification might be open to question in certain cases (Labadie 1984; Labadie 1986). However, the selected plants which were available were collected and their botanical identity was authenticated by three independent experts, as mentioned in the section 'Collection' on p. 121. Fifteen samples of the collected plants were tested for cytotoxicity in vitro. With the described general model of pathogenesis of arbuda in mind (fig.2), two factors for general treatment of arbuda are proposed. Firstly, consolidation of agni is essential (Ca,Ci 15,34). All agnis are regulated by pitta; when pitta increases, agni will also increase. Secondly, there should be a decrease in kapha (Su,Ni 11,21). Kapha and vata are counter-productive: an increase in vata will decrease kapha. Therefore, pitta and vata were selected as main criteria. To know which plants match these criteria, those gunas that increase pitta and vata were selected. Focusing on the mahabhuta composition of the drug, those drugs displaying tejas/agni-properties are given the highest priority, for they increase pitta, followed by those displaying properties of vayu and akasa, for they increase vata. Furthermore, those drugs being katu in taste (rasa) increase both pitta and vata. These drugs generally are katu in vipaka also. Drugs which are madhura, amla and lavana in their rasa should not be used, for they increase kapha. The twenty gunas were categorised according to their proposed priority in treatment of arbuda. Drugs with the properties laghu and suksma can be used, because they increase both pitta and vata. Drugs with tiksna and usna properties can also be used, for they increase pitta. However, drugs which are guru, sthula, manda, sita, picchila, snigdha, sthira, kathina and sandra in their guna should not be used, because they increase kapha and therefore promote arbuda. The virya of the drug should be usna. With these criteria, we made a selection from a list of nearly 500 plant species, used in Ayurveda (Singh/Chunekar 1972; Satyavati/Raina/Sharma 1976; Vaidya 1982; Ojha/Mishra 1985; Satyavati/Gupta/Tandon 1987; Dash 1991; Sharma, P.V. 1992). From the resulting list of 100 species, those which are used traditionally in the treatment of cancer (Manandhar 1980; Malla 1984; Sharma, P.V. 1986; Jain 1991; Jain/DeFillips, 1991; Kirtikar/Basu 1991), and species which were shown previously to have cytotoxic or cytostatic activity (Hegnauer 1962-1990; Dhar/Dhawan/Prasad/Rastogi 1974; Ambasta 1986; Malhotra 1990; Rastogi/Mehrotra, 1990; Rastogi/Mehrotra, 1991) were selected, yielding a list of the following forty-four species:

H.F. Smit, H.J. Woerdenbag, et al., Ayurvedic Herbal Drugs 119 Botanical name Family Sanskrit name Acorus calamus L. Alpinia galanga Willd. Anamirta cocculus Wight. & Arn. Argemone mexicana L. Aristolochia indica L. Araceae Zingiberaceae vaca malayavaca Menispermaceae garalaphala Papaveraceae svarnaksiri Aristolochiaceae isvari Basella rubra L. Bauhinia variegata L. Basellaceae upodika Caesalpiniaceae kancanara Calotropis gigantea (L.) R. Br. Calotropis procera (Ait.) R.Br. Carica papaya L. Carum roxburghianum Kurz. Cleome gynandra L. Colchicum luteum Baker Commiphora mukul (Hook. ex Stocks) Engl. Corallocarpus epigaeus Benth. ex Hook.f. Crataeva nurvala Buch.-Ham. Curcuma longa L. Curcuma zedoaria Rosc. Datura metel L. Delphinium denudatum Wall. Euphorbia neriifolia L. Ficus hispida L.f. Gloriosa superba L. Lagenaria siceraria (Mol.) Standl. Mallotus philippinensis Muell.Arg. Melia azedarach L. Asclepiadaceae alarka Asclepiadaceae arka Caricaceae erandakarkati Apiaceae ajamoda Capparidaceae tilaparni Liliaceae suranjana Burseraceae guggulu Cucurbitaceae sukanasa Capparidaceae varuna Zingiberaceae haridra Zingiberaceae karcura Solanaceae dhattura Ranunculaceae nirvisa Euphorbiaceae snuhi Moraceae kakodumbara Liliaceae langali Cucurbitaceae iksvaku Euphorbiaceae kampillaka Meliaceae Moringa oleifera Lamk. Moringaceae mahanimba sobhanjana Nerium indicum Mill. Apocynaceae karavira Nigella sativa L. Piper betle L. Plumbago zeylanica L. Polyalthia longifolia Thw. Randia uliginosa DC. Ranunculaceae kalajaji Piperaceae tambula Plumbaginaceae citraka Podophyllum hexandrum Royle Berberidaceae vanatrapusi Annonaceae kasthadaru Rubiaceae pinditaka Rhinacanthus nasutus (L.) Kurz. Salvinia cucullata Roxb. Acanthaceae yuthiparni Salviniacee akhukarni Scindapsus officinalis (Roxb.) Schott. Araceae gajapippali Semecarpus anacardium L.f. Anacardiaceae bhallataka

Solanum indicum L. Solanaceae Solanum xanthocarpum Schrad. & Wendl. Solanaceae brhati kantakari Sphaeranthus indicus L. Asteraceae Streblus asper Lour. Moraceae mundi sakhota Urginea indica Kunth. Liliaceae vanapalandu Vitex negundo L. Verbenaceae nirgundi Collection For the collection of the selected plant species, several botanical excursions were undertaken in Kumaon District, Uttar Pradesh, India and Helambu Area, Central Nepal. The methodology used was as follows: selection of the plant, global identification of the plant, global and detailed photography of the plant and its parts for later identification, collection of plant material for the herbarium and for experimental purposes. In the base camp, the collected plants were identified further with the help of local florae (Brandis 1972; Malla/Shrestha/Rajbhandari 1973; Malla 1976; Malla 1984; Malla 1986; Osmaston 1978; Manandhar 1980; Uniyal 1989; Hooker 1990; Naithani 1990; Polunin/Stainton 1990; Stainton 1990; Kirtikar/Basu 1991), and dried in the mild sun. In this way, only some of the selected species could be collected, because of the difficulty of finding specific species, the restriction to seasons in which species could be identified and collected, and the limited amount of available plant material. To overcome these problems, some dried species were obtained from herb markets at Kathmandu and Pathan, both in Nepal, and at Varanasi, India. Rhizomes of Acorus calamus L., fruits of Datura metel L., stems of Plumbago zeylanica L., fruits of Semecarpus anacardium L.f., and fruits of Solanum indicum L. were obtained from the herb market at Kathmandu; rhizomes of Curcuma zedoaria Rosc. were obtained from the herb market at Pathan; and flowers of Calotropis procera (Ait.) R.Br., fruits of Melia azedarach L., roots of Plumbago zeylanica L., fruits of Scindapsus officinalis Schott., and flowers of Sphaeranthus indicus L. were obtained from the herb market at Varanasi. At the campus of the Banaras Hindu University at Varanasi, the bark of Moringa oleifera Lam. was collected and dried. Finally, glands of Mallotus philippinensis Muell. Arg., and fruits of Solanum xanthocarpum Schrad. & Wendl. were obtained from Gorkha Ayurved Company, Gorkha, Nepal. These herbs were destined for the production of Ayurvedic medicines, and appeared to be of good quality. However, when herbs are ob-

H.F. Smit, H.J. Woerdenbag, et al., Ayurvedic Herbal Drugs 121 tained from the market, other problems arise. The identification of the plant material is more difficult, the material is not fresh, and sometimes of inferior quality. Also, substitution and contamination may appear (Labadie 1984; Labadie 1986). To cope with these problems, every collection was guided by an expert of the local flora. More plant species were collected, but only fifteen simplices were of good quality and available in sufficient quantity. The identity of the plants was authenticated by Mr. R.H. Subedi, Gorkha Ayurved Company, Kathmandu, Nepal, Dr. R.R. Koirala, Banaras Hindu University, Varanasi, India, and Dr. M.R. Uniyal, Central Council for Research in Ayurveda and Siddha, New Delhi, India. Voucher specimens are deposited at the Department of Pharmacognosy, University of Utrecht, Netherlands. Botanical name Part IC 50 (mg/ml) Acorus calamus L. rhizoma > 100 Calotropis procera (Ait.) R.Br. flos < 10 Curcuma zedoaria Rosc. rhizoma > 100 Datura metel L. fructus Mallotus philippinensis Muell.Arg glandula Melia azedarach L. fructus > 100 10-100 10-100 Moringa oleifera Lam. cortex > 100 Plumbago zeylanica L. stipes > 100 Plumbago zeylanica L. radix 10-100 Scindapsus officinalis Schott fructus 10-100 Semecarpus anacardium L.f. fructus < 10 Solanum indicum L. fructus > 100 Solanum xanthocarpum Schrad. & Wendl. fructus 10-100 Sphaeranthus indicus L. flos 10-100 Vitex negundo L. folium > 100 cisplatin <10 Figure 3 Cytotoxicity of the extracts of selected plants and the reference compound cisplatin against COLO 320 tumour cells Research The dried material of fourteen plant species³ (fig. 3) was ground and extracted 3 Two simplices were obtained from the same species (Plumbago zeylanica L.).

with 70% ethanol (v/v) using Soxhlet extraction. The extracts were dried, dissolved in ethanol 96%, and diluted in a concentration range. The cytotoxicity of the samples was tested against COLO 320, a human colorectal carcinoma cell line, using the MTT assay (Carmichael/De Graff/Gazdar/Minna/Mitchell 1987). The cell growth inhibition was calculated and the IC 50 value, the drug concentration causing 50% growth inhibition of the tumour cells, was used as a parameter for cytotoxicity (Smit/Woerdenbag/Singh/Meulenbeld/Labadie/ Zwaving 1995). Results The forty-four Ayurvedic plants with potential anti-cancer properties, selected as described in the section 'Material and Methods', are listed on pp. 119 f. From this list, the material of fourteen plants could be collected in India and Nepal. In fig.3 the results of the cytotoxicity tests of the extracts from parts of these fourteen plants are given (Smit/Woerdenbag/Singh/Meulenbeld/ Labadie/Zwaving 1995). Discussion and Conclusion The extracts were prepared with 70% (v/v) ethanol. Although not all possible cytotoxic compounds will be extracted by this method, it might give a representative sample of the plant because a rather broad range of constituents from apolar to polar are extracted with this solvent. Extracts of the flowers of Calotropis procera (Ait.) R.Br. and the nuts of Semecarpus anacardium L.f. displayed the strongest cytotoxic effect, with IC 50 values <? 10 mg/ml. Extracts of several other investigated plants, however, did not show a cytotoxic effect up to 100 µg/ml, the highest concentration tested. In previous experiments with C. procera, extracts of the roots and the leaves showed cytotoxic activity against human epidermal nasopharynx carcinoma (Ayoub/Kingston 1981). The cardenolide calotropin was shown to be responsible for this activity (Kupchan/Knox/Kelsey/Renauld 1964). The cytostatic activity of extracts of S. anacardium has also been described in literature. In one experiment the chloroform extract of the nuts showed an activity of 150% T/C in a P 388 test system in mice, at a dose of 50 mg/kg (Gothoskar/Chitnis/Adwankar/Ranadvie 1971). In another study a fraction of the aqueous methanolic extract of the nuts was tested on Eagles 9 KB nasopharynx carcinoma cell cultures, yielding an IC 50 value of 2.3 uml. This fraction consisted mainly of pentadecylcatechols. These pentadecylcatechols, however, showed no activity on in vivo P 388 leukaemia tests in mice, up to a dose of 80 mg/ml (Hembree/Chang/McLaughlin/Peck/Cassady 1978).

H.F. Smit, H.J. Woerdenbag, et al., Ayurvedic Herbal Drugs 123 As for the other plants tested, limited data on cytotoxicity were found in the literature. Chloroform soluble and insoluble fractions of ethanolic extracts of the whole plant of Solanum indicum L. showed cytotoxicity on seven cancer cell lines: COLO-205 (colon), KB (nasopharynx), HeLa (uterine cervix), HA 22 T (hepatoma), Hep-2 (laryngeal epidermoid), GBM 8401/TSGH (glioma), H 1477 (melanoma). The purified constituents dioscin and methyl protodioscin showed more potent effects by DEA and MTT assay. Dioscin, methyl protoprosapogenin A of dioscin, methyl protodioscin and protodioscin demonstrated cytotoxicity on cultured C 6 glioma cells by PRE assay, and methyl protoprosapogenin A of dioscin, methyl protodioscin and protodioscin showed a tumour inhibitory effect in vivo in C 6 glioma cells. In addition, dioscin showed an inhibitory effect on the DNA synthesis of C 6 glioma cells (Chiang/Tseng/Wang/Chen/Kan 1991). In a two-year study on rats in which the essential oil of the 'Jammu' variety of Acorus calamus L. was given to twenty-five male rats and twentyfive female rats at dietary levels of 0, 500, 1000, 2500 and 5000 ppm, malignant tumours were noted initially after fifty-nine weeks in the duodenal regions of the rats at all levels. Tumours of the same type were not seen in the controls. The oil of the Jammu Calamus, prepared by steam distillation from the rhizomes, contains 75.8% SS-asaron, which was shown to be a carcinogenic compound (Taylor/Jones/Hagan/Gross/Davis/Cook 1967; Stahl/Keller 1981). Anti-cancer and anti-mitotic activity has been reported of plumbagin, the most abundant active principle in the rhizomes of Plumbago zeylanica L. It was shown that plumbagin regresses fibrosarcoma induced by methyl cholanthrene by 70% when given intratumour at a dosage of 2 mg/kg bodyweight in rats (Purushothaman/Mohana/Susan 1983). Our method using data from Ayurveda was shown to provide possible new leads for products which might be useful for the treatment of cancer. Further research on the cytostatic activity of the most active plants, Calotropis procera (Ait.) R.Br. and Semecarpus anacardium L.f. seems interesting. In addition, a more detailed investigation focused on the active compounds is of interest for their molecular nature as well as their mechanisms of action. With the MTT-assay, only a direct toxic effect on the cell can be demonstrated. If the cytotoxic effect is established via an indirect pathway (e.g. by means of the immune system), this is not revealed through the MTT-assay. Acknowledgements We are grateful to the following organisations in the Netherlands for providing funds for this research project: Van Leersum Fund, Groningen; Dr. Hendrik Muller's Vaderlandsch Fonds, The Hague; Foundation Delphi, Groningen; Faculty of Mathe-

H matics and Natural Sciences, University of Groningen, Groningen; Foundation of the Vrij vrouwe van Renswoude, The Hague; Foundation Van der Upwichfonds, Arnhem; Stipendia Fund Royal Dutch Association for the Advancement of Pharmacy (K.N.M.P.), The Hague; Nederlandse Vereniging voor Fytotherapie, Meppel; VSM Geneesmiddelen B.V., Alkmaar. We are also very grateful to the following persons for helping in several aspects in this project: especially Dr. G.J. Meulenbeld, Bedum, The Netherlands, whose valuable advice has been most essential for both the project and this article; further: Dr. H.T. Bakker, Institute of Indian Studies, University of Groningen, The Netherlands; Dr. V.K. Joshi, Department of Dravya Guna, Banaras Hindu University, Varanasi, India; Dr. R.R. Koirala, Varanasi, India; Dr. V.N. Pandey, Central Council for Research in Ayurveda and Siddha, New Delhi, India; Dr. S. Shrestha, Kathmandu, Nepal; Prof. K. Singh, Department of Shalya Shalakya, Gujarat Ayurved University, Jamnagar, India; Prof. L.M. Singh, Kathmandu, Nepal; Mr. R.H. Subedi, Gorkha Ayurved Company, Kathmandu, Nepal; Dr. M.R. Uniyal, Central Council for Research in Ayurveda and Siddha, New Delhi, India; Mrs. Yara Anderson, Department of Pharmaceutical Biology, University of Groningen, The Netherlands. Abbreviations Ah Astangahrdaya Ca Carakasamhita Ci MON Ni Nidanasthana Sa Sarirasthana Su Susrutasamhita Cikitsasthana/Cikitsitasthana Su Sutrasthana Ut Uttarasthana/Uttaratantra References Ambasta, S.P. (1986): The useful plants of India. Delhi, India: Publications & Information Directorate, CSIR. Ayoub, S.M.H.; Kingston, D.G.I. (1981): 'Screening of plants used in Sudan folk medicine for anticancer activity (I).' Fitoterapia 52, 281-284. Bajracharya, M.B. (1987): The Ayurvedic records of cancer treatment. Kathmandu, Nepal: Piyushavarsi Ausadhalaya. Brandis, D. (1972): The Forest Flora of North-West and Central India. Reprint (1874) Dehra Dun, India: Bishen Singh Mahendra Pal Singh. Carmichael, J.; De Graff, W.G.; Gazdar, A.F.; Minna, J.D.; Mitchell, J.B. (1987): 'Evaluation of a tetrazolium-based colorimetric assay: assessment of chemosensitivity testing.' Cancer Research 47, 936-942. Chiang, H.C.; Tseng, T.H.; Wang, C.J.; Chen, C.F.; Kan, W.S. (1991): 'Experimental antitumour agents from Solanum indicum L.' Anticancer Research 11, 1911-1917.

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Summary According to Ayurveda, food and drugs are composed of five basic elements, the pancamahabhutas. Their properties and qualities are expressed in rasa, vipaka, guna, virya, prabhava and karman. In the research programme described in this article, an inventory of Ayurvedic herbal drugs was made, along with their properties and qualities. From this list, we aimed to select plants that would serve as a lead for possible anti-tumour activity. To achieve this, in translation the basic Ayurvedic literature (Carakasamhita, Susrutasamhita and Astangahrdaya) was reviewed for any lead to malignant growths. One syndrome, arbuda, which means 'swelling', was selected to serve as a lead towards possible cancerous diseases. According to the Ayurvedic literature, a model for the pathogenesis of arbuda was made. Based on this, selection criteria were formed; these were used to select plants from the list of Ayurvedic herbal drugs. Some of the selected plant species could be collected in India and Nepal. The dried material of fourteen plants was subjected to ethanol (70% v/v) extraction and the extracts were tested for cytotoxicity against COLO 320 tumour cells, using the microculture tetrazolium (MTT) assay. The IC 50 value - the concentration causing 50% growth inhibition of the tumour cells - was used as a parameter for cytotoxicity. Extracts of the flowers of Calotropis procera (Ait.) R.Br. (Asclepiadaceae) and of the nuts of Semecarpus anacardium L.f. (Anacardiaceae), displayed the strongest cytotoxic effect with IC 50 values < 10 µg/ml. The extracts of several other plants however, did not show any cytotoxic effect up to 100 mg/ml, the highest concentration tested.