Planta Journal of Medicinal Research medica Organ der Editor-in-Chief A. Baerheim-Svendsen, E. Hecker, Heidelberg J. M. Rowson, Mablethorpe Gesellschaft für E. Reinhard, Tübingen Leiden R. Hegnauer, Leiden M. v. Schantz, Helsinki Arzneipflanzen• Pharmazeutisches Institut H. Böhm, Halle W. Herz, Tallahassee K. F. Sewing, Hannover forschung Auf der Morgenstelle 8 F. Bohlmann, Berlin K. Hostettmann, Lausanne E. J. Shellard, London D-7400 Tübingen A. Cave, Chatenay-Malabry H. Inouye, Kyoto S. Shibata, Tokyo P. Delaveau, Paris M. A. Iyengar, Manipal Ch. Tamm, Basel Editorial Board Ding Guang-sheng, F. Kaiser, Mannheim W. S. Woo, Seoul H. P. T. Ammon, Tübingen Shanghai F. H. Kemper, Münster Xiao Pei-gen, Beijing W. Barz, Münster C. -J. Estler, Erlangen W. R. Kukovetz, Graz E. Reinhard, Tübingen N. Farnsworth, Chicago J. Lemli, Leuven •O. Sticher, Zürich H. Floss, Columbus Liang Xiao-tian, Beijing H. Wagner, München H. Friedrich, Münster M. Lounasmaa, Helsinki M. H. Zenk, München D. Fritz, Weihenstephan M. Luckner, Halle A. G. Gonzalez, La Laguna J. Lutomski, Poznan Advisory Board O. R. Gottlieb, Sao Paulo H. Menßen, Köln N. Anand, Lucknow E. Graf, Köln E. Noack, Düsseldorf R. Anton, Strasbourg H. Haas, Mannheim J. D. Phillipson, London

Contents

Volume 49,1983

ISSN 0032-0943 (§)• Hippokrates Hippokrates Verlag Stuttgart II

Contents 49,1983

Atta-ur-Rahman, Bashir, M.\ Isolation of New Cannabinoi'ds in Phelipaea ramosa, a Parasite of Canna- from Catharanthus roseus 124 bis sativa) 250 Atta-ur-Rahman, Nisa, M., Farhi, S.: Isolation of Moenjod-

aramine from Buxus papilosa 126 Galun, £., Aviv, D., Dantes, A.y Freeman, A. \ Biotransfor• mation by Plant Cells Immobilized in Cross-Linked Poly- Balsevich, J., Kurz, W. G. W.: The Role of 9- and/or 10- acrylamide-Hydrazide. Monoterpene Reduction by En- oxygenated Derivatives of Geraniol, Geranial, Nerol, trapped Mentha Cells 9 and Neral in the Biosynthesis of Loganin and Ajmalicine 79 Ghosal, Sh., Singh, A.K., Biswas, K.: New6-Aryl-2-pyrones Bassleer, R., Marnette, J.-M., Wiliquet, Ph., De Pauw-Gillet, from Gentianapedicellata 240 M.-CL, Caprasse, M., Angenot, L.\ Etüde complemen-

taire de la cytotoxicite de la melinonine F, alcaloide derive Hafez, A.y Adolf, W., Hecker, E.\ Active Principles of the de la ß-carboline (Complementary Study of Cytotoxic Thymelaeaceae. III. Skin Irritant and Cocarcinogenic Activity of Melinonine F) 158 Factors from Pimelea simplex 3 Becker, H., Chavadej, S., Weberling, F.: Valepotriates in Valeriana thalictroides 64 leven, M., van den Berghe, D. A., Vlietinck, A. J.: Plant Becker, H., Herold, S.: RP-8 als Hilfsphase zur Akkumula• Antiviral Agents. IV. Influence of Lycorine on Growth tion von Valepotriaten aus Zellsuspensionskulturen von Pattern of Three Animal Viruses 109 Valeriana wallichii (RP-8 Auxiliary Phase for the Accu- Ishiguro, K., Yamaki, A/., Takagi, 5.: Studies on Iridoid- mulation of Valepotriates from Cell-Suspension-Culture related Compounds; III: Gentiopicral, the Aglucone of of Valeriana wallichii) 191 Gentiopicroside 208 Bounthanh, C, Richert, L., Beck, J. P., Haag-Berrurier, M., Anton, R.: The Action of Valepotriates on the Synthesis Jakovlev, V., Isaac, O., Flaskamp, E.: Pharmakologische of DNA and Proteins ofCultured Hepatoma Cells. . . . 138 Untersuchungen von Kamillen-Inhaltsstoffen, VI. Unter• Brianqon-Scheid, F., Lobstein-Guth, A., Anton, R.: HPLC suchungen zur antiphlogistischen Wirkung von Chama- Separation and Quantitative Determination of Biflavones zulen und Matricin (Pharmacological Investigations with in Leaves from Ginkgo biloba 204 Compounds of Chamomile, VI. Investigations on the Antiphlogistic Effects of Chamazulene and Matricine) 67 Caputo, O,, Delprino, L., Viola, F., Caramiello, R., Bal- Joshi, K. C, Singh, P., Taneja, S.: A Sesquiterpenoid Naph- liano, G.: Biosynthesis of Sterols and Triterpenoids in thol from Kydia calycina 127 Tissue Cultures of Cucurbita maxima 176 Jossang, A., Leboeuf, M., Cabalion, P., Cave, A.: Alcaloides Chagnon, M., Ndibwami, A., Dube, S., Bumaya, Ar. Ac- des Annonacees. XLV: Alcaloides de Polyalthia niti- tivite Anti-Inflammatoire d'Extraits de Crassocephalum dissima (Alkaloids from Annonaceae. XLV: Alkaloids multicorymbosum (Anti-inflammatory Action of an Ex- of Polyalthia nitidissima) 20 tract from Crassocephalum multicorymbosum) 255 Chattopadhyay, S., Chattopadhyay, U., Mathur, P. P., Saini, Kimura, Y., Ohminami, H., Okuda, H., Baba, K., Kozawa, K. S., Ghosal, S.: Effects of Hippadine, an Amarylli- M., Arichi, S.: Effects of Stilbene Components of Roots daceae , on Testicular Function in Rats 252 of Polygonum ssp. on Liver Injury in Peroxidized Oil-fed Chen Weiming, Yan Yaping, Liang Xiaotian: Alkaloids from Rats 51 Roots of A Istonia yunnanensis 62 Kisiel, W.: 8-Epidesacylcynaropicrin from Crepis capillaris 246 Kiso, Y., Suzuki, Y., Watanabe, N., Oshima, Y., Hikino, H.: Dehaussy, H., Tits, M., Angenot, L. \ La guattegaumerine, Antihepatotoxic Principles of Curcuma longa Rhizomes 185 nouvel alcaloide bisbenzylisoquinoleinique de Guatteria Kiso, Y., Tohkin, M., Hikino, H.\ Assay Method for Anti• gaumeri (Guattegaumerine, New Bisbenzylisoquinoline hepatotoxic Activity Using Carbon Tetrachloride In- Alkaloid from Guatteria gaumeri) 25 duced Cytotoxicity in Primary Cultured Hepatocytes . . 222

Dominguez, X. A., Franco, R., Cano, G., Ma. Consue lo Kram, G.f Franz, G.: Untersuchungen über die Schleim- Garcia, F., Xorge A, Dominguez, S. Jr., Leonardo de la polysaccharide aus Lindenblüten (Analysis of Linden Pena,M.: Isolation of a New Furano-l,4-Naphthaquinone, Flower Mucilage) 149 Diodantunezone from Lanthana achyranthifolia .... 63 Krüger, £>., Junior, P., Wichtl, M.\ Neue Cardenolidglyko- side aus Digitalis lanata (New Cardiac Glycosides from

Endo, K., Oshima, Y., Kikuchi, H., Koshihara, Y.} Hikino, Digitalis lanata) 74 H.: Hypotensive Principles of Uncaria Hooks 188 Engelshowe, R.: Dimere Proanthocyanidine als Gerbstoff• Langhammer, L., Schulze, G., Gujer, R., Magnolato, D., vorstufen in Juniperus communis (Tannin Producing Horisberger, M.\ Isolation and Structure of a Rarely DimericProanthocyanidins in Juniperus communis). . . 170 Occurring Cyanidanol Glycoside from Cortex Betulae . 181 Esterl, A., Gab, S., Bieniek, D.\ Zur Kenntnis der Inhalts• Lemli, J., Cuveele, /., Verhaeren, E.\ Chemical Identification stoffe von Isertia hypoleuca (On The Constituents of of Alexandrian and Tinnevelly Senna. Studies in the Isertia hypoleuca) 244 Field of Drugs Containing Anthracene Derivatives XXXIV 36

Fournier, G., Paris, M.: Mise en Evidence de Cannabinoides Long-Ze Lin, Wagner, H.t Seligmann, ö.\ Thalifaberine, Chez Phelipaea ramosa, Orobanchacees, Parasitant le Thalifabine and Huangshanine, Three New Dimeric Chanvre, Cannabis sativa, Cannabinacees (Detection of Aporphine-Benzylisoquinoline Alkaloids 55 Contents III

Man-Po Wong, Teh-Chang Chiang, Hson-Mou Chang: Chem• Rueffer, M., Nagakura, N., Zenk, M. H.: Partial Purification ical Studies on Dangshen, the Root of Codonopsis pilo- and Properties of S-Adenosylmethionine: (R), (S)-Nor- sula 60 laudanosoline-6-O-Methyltransferase from Misawa, M., Hayashi, M., Takayama, S.: Production of platyceras Cell Cultures 131 Antineoplastic Agents by Plant Tissue Cultures. I. In- duction of Callus Tissues and Detection of the Agents in Sariyar, G.: Alkaloids from triniifolium of Turkish Cultured Cells 115 Origin 43 Seip, E. H., Ott, H. H., Hecker, E.: Skin Irritant and Tumor Nahrstedt, A., Wray, V., G rot Jahn, L., Fikenscher, L. H., Promoting Diterpene Esters of the Tigliane Type from Hegnauer, R.: New Acylated Cyanogenic Diglycosides the Chinese Tallow Tree (Sapium sebiferum) 199 from Fruits of Anthemis cairica 143 Sepulveda-Boza, S., Friedrichs, £., Puff, H., Breitmaier, E.\

Nomura, T., Fukai, T.} Shimada, T., Ih-Sheng Chen: Com- Ein iso-Homoprotoberberin-Alkaloid aus den Wurzeln ponents of Root Bark of Morus australis. I. Structure of von Berberis actinacantha (An iso-Homoprotoberberin- a New 2-Arylbenzofuran Derivative, Mulberrofuran D. 90 Alkaloid from the Roots of Berberis actinacantha).... 32 Shoyama, Y., Hatano, K., Nishioka, I.: Clonal Multiplica- Obata-Sasamoto, H., Komamine, A.: Effect of Culture Con- tion of Pinellia ternata by Tissue Culture 14 ditions on DOPA Accumulation in a Callus Culture of Stoianova-Ivanova, B., Budzikiewicz, H., Koumanova, B., Stizolobium hassjoo 120 Tsoutsoulova, A., Mladenova, K., Brauner, A.: Essential Ohiri, F. C, Verpoorte, R., Baerheim Svendsen, A.: !H NMR Oil of Chrysanthemum indicum 236 Chemical Shift Values for Aromatic Protons in 2,3,9,10- and 2,3,10,11-tetrasubstituted Tetrahydroprotoberberine Teh-Chang Chiang, Hson-Mou Chang, Mak, Th. C. W.: New Alkaloids 162 Oleanene-type Triterpenes from Abrus precatorius and Ohiri, F. C, Verpoorte, R., Baerheim Svendsen, /i.:Tertiary X-ray Crystal Structure of Abrusgenic Acid-Methanol Phenolic Alkaloids from Chasmanthera dependens ... 17 1:1 Solvate 165 Ojewole, J. A. O., Adesina, S. K.: Mechanism of the Hypo- tensive Effect of Scopoletin Isolated from the Fruit of Tetrapleura tetraptera 46 van der Sluis, W. G., van der Nat, J. M., Spek, A. L., Ike- shiro, Y., Labadie, R. P.: Gentiogenal, a Conversion Pro- Ojewole, J. A. O., Adesina, S. K.: Cardiovascular and Neuro- duct of Gentiopicrin (Gentiopicroside) 211 muscular Actions of Scopoletin from Fruit of Tetrapleura tetraptera 99 Okogun, J. L, Adeboye, J. O., Okorie, D. A. \ Novel Struc- Wagner, H., Schwarting, G., Varljen, Bauer, R., Ham- tures of two Chromone Alkaloids from Root-Bark of dard, M. E., El-Faer, M. Z., Beal, J.\ Die chemische Zu• Schumanniophyton magnificum 95 sammensetzung der Convolvulaceen Harze IV. Die Gly- kosidsäuren von Ipomoea quamoclit, I. lacunosa, I. pan- Palacios, P., Gutkind, G., Rondina, R. V. D., de Torres, R., durata und Convolvulus al-sirensis (Chemical Consti• Coussio, J. D.: Genus Baccharis. II. Antimicrobial Activ• tuents of the Convolvulaceae-Resins IV. The Glycosidic ity of B. crispa and B. notosergila 128 Acids of Ipomoea quamoclit, I. lacunosa, 1. pandurata Pandey, V. B., Singh, J. P., Mattocks, A. R., Bailey, E.\ A and Convolvulus al-sirensis) 154 Note on "Isolation and Pharmacological Action of Helio- Watanabe, K, Watanabe, H., Goto, Y., Yamaguchi, M., trine, the Major Alkaloid of Heliotropium indicum Seeds" 254 Yamamoto, N., Hagino, K.\ Pharmacological Properties Pathak, V. P., Saini, T. R., Khanna, R. N.: A New Furano- of Magnolol and Hönokiol Extracted from Magnolia offi- flavone from Seeds of Pongamia glabra 61 cinalis: Central Depressant Effects 103 Perera, P., Sandberg, F., van Beek, T. A., Verpoorte, R.: Willuhn, G., Röttger, P.-M., Matthiessen, U.: Helenalin- Tertiary Alkaloids of dicho- und 11,13-Dihydrohelenalinester aus Blüten von Arnica toma Seeds 28 montana (Helenalin- and 11,13-Dihydrohelenalinester Perera, P., van Beek, T. A., Verpoorte, R.\ Dichomine, a from Flowers of Arnica montana) 226 Novel Type of Iboga Alkaloid 232 Witte, L., Berlin, J., Wray, V., Schubert, W., Kohl, W., Höfle, G., Hammer, J.: Mono- and Diterpenes from Cell Ravid, U., Putievsky, E.: Constituents of Essential Oilsfrom Cultures of Thuja occidentalis 216 Majorana syriaca, Coridothymus capitatus and Satureja thymbra 248 Yang Minghe, Chen Yanyong: Steroidal Sapogenins in Dios- Röder, E., Wiedenfeld, H., Honig, A.: Pyrrolizidinalkaloide corea collettii 38 aus Senecio aureus 57 Yu De-Quan, Das, B. C.: Structure of Hydroxymuscopyri- Rueffer, M., Nagakura, N., Zenk, M. H: A Highly Specific O- dine A and Hydroxymuscopyridine B, Two New Constit• Methyltransferase for Nororientaline Synthesis Isolated uents ofMusk 183 from Argemoneplatyceras Cell Cultures 196 Yu De-quan, Y., Das, B. C.: Alkaloids of Aconitum barbatum 85 IV

Subject Index 49,1983

A Cocarcinogens 3 Gentiogenal 211 Convolvulaceae resins 154 Gentiopicral 208 Abietane 220 Coreximine 14 Gentiopicrin 211 Abrusgenic acid 165 28 Gentiopicroside 208, 211 Abruslactone 165 Coumaric acid 186 Geranial 79 Acevaltrate 64, 191 Coumarins 46, 99 Geraniol 79 Adaptogenes 240 Cucurbitacin 176 Ginkgetin 204 Alkaloids 17, 20, 25, 28, 32, 43, 55, 57, 62, Curcumin 185 Glycosidic acids 154 79, 85, 95,109,124, 126,131,158,162, Curcuminoids 185 Glycyrrhetinic acid 224 188,196, 232, 244, 252, 254 Cyanogenic glycosides 143 Glycyrrhizin 224 Amyrin 177 Cycloartenol 177 Govanine 17 Antineoplastic agents 115 Cymenol 216 Guajazulene 67 Antiviral agents 109 Cynarin 224 Guattegaumerine 25 128 Cynaropicrin 246 Aporphine alkaloids 20 Cytotoxic action 158 Aporphine derivatives 43 H Apparicine 232 Arabinose 149 D Helenalin 226 Arylbenzofuran derivatives 90 Helenalinester 226 Daphnane derivatives 3 Avenasterol 177 Helenanolides 226 Dauricine 20 Heliotrine 254 Daurisoline 20 Hepatoma cells 138 Dehydrohelenalinester 226 B Hibiscone 127 Deoxyphorbol 199 Hibiscoquinone 127 Dichomine 232 Benzylisoquinoline alkaloids 131 Hinikione 220 Didrovaltrate 64, 138, 191 Berberine alkaloids 32 Hinokiol 220 Diginatigenin derivatives 74 Biflavones 204 Hippadine 252 Digoxigenin derivatives 74 Bilobetin 204 Homovaltrate 64, 191 Dihydrocadambine 188 Honokiol 103 Biosynthesis of alkaloids 131, 196 Dihydroquinamine 244 Huangshanine 55 Biosynthesis of ajmalicine 79 Diodantunezone 63 Hydroxymuscopyridine 183 Biosynthesis of loganin 79 Diosgenin 38 Hydroxymusizin glycoside 36 Biosynthesis of sterols 176 Diterpene esters 3, 199 Biosynthesis of triterpenoids 176 Diterpenes 216 Biotransformation 9 Diterpenoid alkaloids 85 Bisabolol 67 Dopa 120 Bisbenzylisoquinoline alkaloids 20, 25, 55 Iboga alkaloids 232 Bisnorargemonine 14 28 Borneol 236 Iboxygaine 232 Bornyl acetate 236 Immobilized cells 9 Enzymes 196 Indicine 254 Epilumicin 143 glucoside 188 C Epoxyphorbol 199 Indole alkaloids 28, 62, 79, 124. 158, 188, Erythrocentaurin 212 232 Cadambine 188 Essential oils 9, 67, 216, 236, 248 Iridoids 208,211 Caffeic acid 186 Isochinoline alkaloids 196 Camphor 216 Isodihydrocadambine 188 Cannabinoi'ds 250 Isoginkgetin 204 Cardenolides 74 Isoquinoline alkaloids 32 Carvacrol 216 Fenchone 220 Isothujone 220 181 Ferruginol 220 Isovaltrate 64, 191 Catechinxylopyranoside 181 Ferulic acid 186 Cell cultures 9, 14 79, 115, 120, 131, 176, Flavonoids 61, 63, 90, 128, 204 191,196,216 Fluorocarpamine 124 K Chamazulene 67 Fridelin 60 Cholestanol 177 Furoflavones 61 Kuwanon 90 Chromone alkaloids 95 Chrysanthenone 236 Chrysosplenetin 63 G L Cinchonamine 244 Cinnamic acid 186 Galactose 149 Lasiocarpine 254 Clonal multiplication 14 Genkwanin 128 Laudanosoline 196 Subject Index V

Levomenol 67 Pedicellin 240 Steroidal sapogenins 38 Liliolide 240 Penduletin 63 Stigmastenol 177 Lindolhamine 20 Perakine 62 Stilbene51 Linoleic acid 220 Perivine 232 Sugiol 220 Liriodenine 20 Phorbol 199 Swertiamarin 240 Lumicin 143 Phytosterols 60, 176 Lycorine 109 Piceid 51 Picroside 224 Pimelea factors 3 M Pleiocarpamine 124 Tabersonine 28 Podophyllotoxin 224 Tacaman alkaloids 232 Magnolol 103 Polysaccharides 149 Tannins 181 Mandelonitril glycosides 143 Proanthocyanidins 170 Taraxerol 60 Matricine 67 Protoberberin derivatives 32 Terpineol 216 Melionine 158 Protoberberine alkaloids 20 Tertiary phenolic alkaloids 14 Mentenediol 220 Protocatechuic acid 186 Tetrahydroalstonine 62 Menthone 9 Pseudoguaianolides 226, 246 Tetrahydroisochinoline alkaloids 196 Methionine 224 Puberaconitidine 85 Tetrahydroprotoberberine alkaloids 162 Methylabrusgenat 165 Puberaconitine 85 Thalifaberine 55 Methylnorlaudanosoline 196 Puberanidine 85 Thalifabine 55 Methyltransferase 131, 196 Puberanine 85 Thujaplicin derivatives 216 Moenjodaramine 126 Pulegone 9 Thujone 220 Monoterpenes 216 Pyrrolizidine alkaloids 57 Tigliane derivatives 3 Morphinane derivatives 43 Tinnevellin glycoside 36 Morusin 90 Tripdiolide 109 Mucilage 149 Triterpenoids 60, 165 Myrtanol 216 Tropolone 220 Tumor promotors 199 Resins 154 Resveratrol 51 N Reticuline 20, 131 U Rhamnose 149 Naphthalene glycosides 36 Rhoeadine derivatives 43 Ursolic acid 240 Naphthaquinones 63 Ushinsunine 20 Neolignane derivatives 103 Neomenthol 9 S Neral 79 V Nerol 79 Sabinene 220 Norlaudanosoline 131 Valepotriates 64, 138, 191 Sapogenins 38 Normacusine 62 Valtrate 64, 138,191 Saponins 38, 60 Nororientaline 196 Vellosimine 62 Sarpagine 62 Norreticuline 131 Vindolinine derivatives 124 Schumannificine 95 Vinorine 62 Sciadopitysin 204 Voacangine 28 Scopoletin 46, 99 O Voacristine 232 Secoiridoids 208,211 Voaphylline 28,232 Sesquiterpene lactones 226,246 Vobasine 232 alkaloids 131 Sesquiterpenes 127 Orientaline 196 Silybin 224 Simplexin 3 Y Sinapic acid 186 P Sitosterol 220 Skin irritants 199 Yamogenin 38 Pallidine 14 Spinasterol 177 Palmatic acid 220 Stemmadenine 28 Z Papaver alkaloids 43 Stepholidine 20 Pedicellatin 240 Steroidal alkaloids 126 Zierinxyloside 143 VI

Index of Names of Organisms 49,1983

Please note that the following plant fami- lies are listed as indicated below: Alsinaceae sub Caryophyllaceae Cacalia floridana 57 Fagara zanthoxyloides 116 Apiaceae sub Umbelliferae Caesalpinia gilliesii 116 Fumaria officinalis 133 Arecaceae sub Palmae Caesalpiniaceae 36 Asteraceae sub Compositae Campanulaceae 60 Brassicaceae sub Cruciferae Cannabis sativa 250 Clusiaceae sub Guttiferae Cassia angustifolia 36 Fabaceae sub Leguminosae (including Mi- Cassia senna 36 Gentiana pedicellata 240 mosoideae = Mimosaceae, Caesalpini- Catharanthus roseus 79,124 Gentianaceae 208, 211, 240 oideae = Caesalpiniaceae and Papilion- Celastraceae 115 Ginkgo biloba 204 oideae = Papilionaceae = Fabaceae Cephalotaxaceae 115 Ginkgoaceae 204 sensu stricto) Cephalotaxus harringtonia 115 Glaucium flavum 133 Hypericaceae sub Guttiferae Chamomilla recutita 67 Guatteria gaumeri 25 Lamiaceae sub Labiatae Chasmanthera dependens 17,162 Oenotheraceae sub Onagraceae Chelidonium majus 133 Poaceae sub Gramineae Chlorella ellipsoidea 178 H Chrysanthemum indicum 236 Cissampelos mucronata 133 Heliotropium indicum 116,254 Clivia miniata 109 Heliotropium supinum 254 Codonopsis pilosula 60 Hibiscus species 127 Abrus cantoniensis 165 Colchicum speciosum 116 Holacantha emoryi 116 Abrus precatorius 165 Compositae 57,67,128,143,226,236,246, Hura crepitans 3 Aconitum barbatum 85 255 Adlumia fungosa 133 Convolvulaceae 154 Albertisia papuana 22 Convolvulus al-sirensis 154 I Alstonia yunnanensis 62 Convolvulus microphyllus 154 Althaea officinalis 152 Coridothymus capitatus 248 Ipomoea lacunosa 154 Amaryllidaceae 109, 252 Corydalis pallida 133 Ipomoea operculata 154 Annonaceae 20, 25 Corydalis sempervirens 133 Ipomoea pandurata 154 Anthemis altissima 143 Crassocephalum multicorymbosum 255 Ipomoea quamoclit 154 Anthemis cairica 143 Crepis capillaris 246 Ipomoea turpethum 154 Anthocephalus cadamba 189 Crepis virens 246 Isertia hypoleuca 244 Apocynaceae 28, 62, 79,124, 232 Crinum asiaticum 252 Araceae 14 Crinum augustum 252 Argemone intermedia 133 Crinum latifolium 252 Argemone platyceras 131, 196 Crinum pratens 252 Arnica montana 226 Cucumis sativus 176 Juniperus communis 170 Artemisia caruthii 67 Cucurbita maxima 176 Cucurbita pepo 177 Cucurbitaceae 176 B Cupressaceae 170, 216 Curcuma longa 185 Kydia calycina 127 Baccharis crispa 128 Baccharis megapotamica 116 Baccharis notosergila 128 Baliospermum montanum 200 D Berberidaceae 32,133 Labiatae 9, 248 Berberis actinacantha 32 Dictiostelium discoideum 177 Lantana achyranthifolia 63 Berberis henryana 133 Digitalis lanata 74 Leguminosae 36, 46, 61, 99, 120,165 Berberis stolonifera 133 Digitalis schischkinii 74 Lindera oldhamii 22, 26 Berberis wilsonae 133 Dioscorea collettii 38 Loganiaceae 158 Betula species 181 Dioscoreaceae 38 Betulaceae 181 Blackstonia perfoliata 211 M Bombax malabaricum 127 Boraginaceae 254 Magnolia obovata 103 Brucea antidysenterica 116 Magnolia officinalis 103 Bryonia dioica 176 Erythrina lithosperma 22 Magnoliaceae 103 Buxaceae 126 Eschscholtzia tenuifolia 133 Majorana syriaca 248 Buxus papulosa 126 Euphorbiaceae 199 Malvaceae 127 Index of Names of Organisms Vll

Maytenus bucchananii 116 Penicillium expansum 211 Strychnos melinoniana 158 Menispermaceae 17, 133, 162 Phelipaea ramosa 250 Strychnos usambarensis 158 Menispermum dauricum 22 Pimelea linifolia 7 Mentha species 9 Pimelea prostrata 3 Mimosaceae 46, 99 Pimelea simplex 3 T Moraceae 90 Pinellia ternata 14 Morus alba 90 Podophyllum hexandrum 223 Tabernaemontana dichotoma 28, 232 Morus australis 90 Polyalthia beccarii 20 Tabernaemontana eglandulosa 232 Moschus moschiferus .183 Polyalthia emarginata 20 Taxus brevifolia 116 Mycobacterium species 10 Polyalthia nitidissima 20 Tetrapleura tetraptera 46, 99 Polyalthia oligosperma 20 Thalictrum dasycarpum 116 Polyalthia oliveri 20 Thalictrum faberi 55 N Polyalthia suaveolens 20 Thalictrum sparsiflorum 133 Polygonaceae 51 Thalictrum tuberosum 133 Nicotiana tabacum 250 Polygonum cuspidatum 51 Thuja occidentalis 216 Polygonum multiflorum 51 Thymelaeaceae 3 Polypodium vulgare 181 Thymus capitatus 248 O Pongamia glabra 61 Tilia species 149 Putterlickia verrucosa 115 Tiliaceae 149 Ochrosia moorei 116 Tripterygium wilfordii 115 Origanum compactum 249 Origanum floribundum 249 R Origanum hirtum 249 U Origanum majorana 249 Ranunculaceae 55, 85,133 Origanum maru 248 Rhus coriaria 248 Ulmus americana 181 Origanum smyrnaeum 249 Rubiaceae 95,188,244 Uncaria sinensis 188 Origanum syriacum 248 Uncaria species 188 Orobanchaceae 250 S

V P Sapium sebiferum 199 Satureja capitata 248 Panax ginseng 60 Satureja peltieri 249 Valeriana edulis ssp. procera 64 Panax pseudo-ginseng 60 Satureja thymbra 248 Valeriana kilimandscharica 64 Papaver armeniacum 43 Schistosoma japonicum 199 Valeriana thalictroides 64 Papaver cylindricum 43 Schumanniophyton magnificum 95 Valeriana wallichii 138,191 Papaver fugax 43 Schumanniophyton problematicum 97 Valerianaceae 64, 138, 191 Papaver Orientale 196 Scolytus multistriatus 181 Verbenaceae 63 131,196 Scolytus ratzeburgi 181 Papaver tauricola 43 Scrophulariaceae 74 Papaver triniifolium 43 Senecio aureus 57 Z 43,131,196 Stizolobium hassjoo 120 Papilionaceae 120 Streptomyces clavuligerus 10 Zingiberaceae 185 VIII

Pharmacology Index 49,1983

Biological Systems Pharmacological Effects/ Plant/Constituent Page Organs / Diseases Effects on

Central nervous System anticonvulsive Magnolia officinalis, Magnoliaceae 103 muscle relaxant Magnolia officinalis 103 sedative Magnolia officinalis 103 Peripheral nervous System muscle relaxant Tetrapleura tetraptera, Mimosaceae 99 Autonomous nervous System spasmolytic Tabernaemontana dichotoma, Apocynaceae 28 Cardiovascular System hypotensive Tetrapleura tetraptera, Mimosaceae 46 hypotensive Tetrapleura tetraptera, Mimosaceae 99 Hormonal System antifertility Hippadine, Amaryllidaceae 252 Infectious deseases antibacterial Baccharis crispa, Asteraceae 128 antiviral Clivia miniata, Amaryllidaceae 109 Tumors tumor promotion Pimelea simplex, Thymelaeaceae 3 Inflammation antiinflammatoric Chamomilla reculita, Asteraceae 67 antiinflammatoric Crassocephalum multicorymbosum 255 Journal of Planta MoHirinal •••"IUI 1983, Vol. 49, pp. 196-198, © Hippokrates Verlag GmbH Plant Re^aSI medO

A Highly Specific O-Methyltransferase for Nororientaline Synthesis Isolated from Argemone platyceras Cell Cultures

M. Rueffer*, N. Nagakura** and M. H. Zenk*

* Lehrstuhl für Pharmazeutische Biologie der Universität München, Federal Republic of Germany ** Kobe Women's College of Pharmacy, Kobe 658, Japan

Received: August 8,1983

Key Word Index: been isolated from opium poppy [3] and is metaboli- zed in Papaver Orientale via orientalinone to isothe- Argemone platyceras] Papaveraceae; Cell Suspen• baine [4]. Therefore nororientaline can be consid- sion cultures; Alkaloid biosynthesis; 6-O-methyl- ered as a specific precursor of a number of isoquinoli- norlaudanosoline)-5' -O-methyltransferase. ne alkaloids in higher . This report describes the partial purification and characterization of a new enzyme which was designated S-adenosyl-L-methio- Abstract nine: (6-0-methyl-norlaudanosoline)-5'-0-methyl- transferase. A highly specific new enzyme, S-adenosyl-L-me- thionine: (6-O-methyl-norlaudanosoline)-5; -O-me- Material and Methods thyltransferase which catalyses the formation of nor• Argemone platyceras cells were cultivated as described pre• orientaline from 6-O-methyl-norlaudanosoline and viously [2]. Chemicals used were essentially those described in the SAM was discovered, partially purified, and charac- preceeding communication [2]. The new enzyme, described here, terized. Argemone platyceras cell Suspension cultures was purified through Steps 1-4 as described previously [2]. How- ever, the enzyme was eiuted from hydroxyapatite columns at frac- served as enzyme source. tions 56-65 in contrast to the previously described NLS-6-O-me- thyltransferase which was eiuted between fractions 43 and 54. En• zyme assays were carried out as follows: (R, S)-6-0-methyl-nor- laudanosoline (0.3 mM), SAM-3H (10000 cpm, 0.1 mM), KP0 2- Introduction 4 buffer, pH 7.5 (130 mM) were incubated in a total volume of 150 ul at 35° C for 45 min. The reaction was terminated by the addition of Tetrahydroisoquinoline biosynthesis in plants in- 200 ul Na2C03-buffer (1 M, pH 9.5). The methylated product was volves a number of O- and N-methylation Steps [e.g. extracted by adding 400 ul isoamylacohol and shaking for 45 min. 1] of norlaudanosoline, the first precursor in the After centrifugation for 5 min. in an Eppendorf centrifuge, 200 ul pathway. In a previous communication, we have des• of the organic phase were transferred to scintillation vials and counted. A blank value of about 10 % (obtained by assay mixtures cribed the isolation of a first specific O-methyltrans- containing either no enzyme or no Substrate), had to be subtracted ferase. In Argemone platyceras cell Suspension cultu• from all incubation mixtures. Recovery of thje methylated product res this enzyme methylates predominantly the 6-0- was 92 % under these conditions. position of (S)- as well as (R)-norlaudanosoline, me- The products were separated and identified by HPLC using a diating the transfer of a methyl-group from S-adeno- Nucleosil-SA-column (25 mm x 3.2 mm i.d.) and 0.5 M ammo- nium phosphate : methanol (70:30) as a solvent System. Retention syl-L-methionine to NLS [2]. Düring the purification time of the potential products were: norisoorientaline, 7.14 min.; of this enzyme, a second enzymatic activity was dis• nororientaline, 8.20 min.; norprotosinomenine, 8.84 min.; norre- covered which utilises 6-O-methyl-NLS to introduce ticuline, 10.94 min. a second methyl group in the 5;-position of the mole- Preparative isoiations were done using the above incubation mixture x 100. The reaction product was extracted by ethylacetate cule to yield nororientaline. Again SAM serves as a and purified twice by thin layer chromatography (Si-gel; solvent methyl group donator. Nororientaline is a principal Systems: 1) Chloroform : methanol : acetic acid : water = immediate precursor of papaverine in Papaver som• 18:6:3:0.3; 2) acetone : Chloroform : diethylamine = 5:4:1). Mass niferum [1] and is also undoubtedly converted to spectra were determined in a Finnigan MAT 44S instrument. orientaline by N-methylation. Orientaline itself has Results and Discussion

Abbreviations: NLS = Norlaudanosoline; SAM = S-Adeno- Düring the purification of the 6-O-methyltransfe- syl-L-methionine; SAH = S-Adenosylhomocysteine. rase evidence was obtained that its reaction product O-Methyltransferase for Nororientaline Synthesis 197 Table I Purification protocol for the ö'-O-methyltransferase from A. platyceras cell Suspension cultures (125 g fwt)

Purification step Volume Total activity Protein Spec. activity Yield Purification (ml) (pkat) (mg/ml) (pkat/mg) factor

Crude extract 236 4647 1.10 17.97 1 1.4 (NH4)2S04 precipitation 23 5733 10.02 24.9 100 GelTiltration 69 5703 0.22 375.5 99.48 20.9 DEAE chromatography 35 1127 0.07 506.5 19.65 28.2 Hydroxyapatite 12 327.7 0.038 718.8 5.7 40.0

Table II Table III Some characteristics of (6-0-methylnorlaudanosoline)-5'-0- Substrate specifity of 40-fold purified 5'-0-methyltransferase methyltransferase from A. platyceras cell cultures using SAM as methyl group donator

Characteristics 5'-0-methyltransferase Substrate pkat/mg % Product of reaction Molecular weight 47.000 pH-optimum 7.5 (R, S)-6-0-methyl- Temperature Optimum 35° C norlaudanosoline 314 100 Nororientaline K for 6-O-methylnorlaudano- (R)-Norlaudanosoline 0 0 M - soline 0.4 mM (S)-Norlaudanosoline 0 0 - KM for SAM 0.05 mM (R, S)-Laudanosoline 0 0 Kj for SAH 0.034 mM -

is transformed by the introduction of a second methyl ted for the unspecific meta and para directing cate- group in the presence of excess SAM and crude en• chol-O-methyltransferases from plant origin [5]. The zyme preparations. The two enzymes involved are general properties of the enzyme, described here, are clearly separated by chromatography on hydroxya• very similar to the previously described norlaudano- patite. The enzyme was partially purified by using soline-6-O-methy ltransferase. (R,S)-6-0-methyl-norlaudanosoline as Substrate. As The Substrate specifity of the 5'-0-methyltransfe- shown in Table I the enzyme could be recovered with rase was of considerable interest. 18 Substrates were 6 % yield. Purification was about 40-fold. tested for their ability to accept methyl groups from The characteristic properties of the enzyme are li- SAM catalysed by S-adenosyl-L-methionine: (6-0- sted in Table II. The enzyme has a half life of 33 hrs methyl-norlaudanosoline)-5'-0-methyltransferase. at 30° C and retains füll enzymatic activity in the pres• None of the phenols, phenylpropanoids, flavonoids,

ence of 0.05 % NaN3 at 4° C after 4 weeks. The en• coumarins, and biogenic amines tested, served as zyme is, however, completely inactivated by freezing Substrates. Out of 12 different isoquinoline alkaloids even in the presence of 30 % glycerol. The enzyme is tested [2] only (R,S)-6-0-methyl-norlaudanosoline inhibited completely by Fe3+ and Hg2+ ions at 5 mM was methylated. Surprisingly neither (R) or (S)-nor- concentration and is strongly inhibited by 5 mM p- laudanosoline nor (R, S)-laudanosoline served as a chloromercuribenzoate (84 %), N-ethylmaleimide Substrate (Table III) which shows a hitherto unique (82 %), and 10mM jodobenzoic acid (79 %). This in- Substrate specifity of an O-methyltransferase in plant dicates that SH-groups are present at the active Cen• kingdom [6]. Since the 6-O-methyl-norlaudanosoline ter of the enzyme. Similar findings have been repor- used was a racemate, it is not known whether either

+ SAM + SAH

6-0-Methylnorlaudanosoline Nororientaline 198 Rueffer, Nagakura, Zenk

of the two stereoisomers is preferentially methyl• ated. As shown by HPLC (retention time 8.20 min.) Hippokrates and mass spectroscopy ([M + l]+ 316, 100%, meas- ured with dci), the reaction product of the enzyme was unequivocally nororientaline. As shown in Fig. 1, the new enzyme therefore exclusively methylates the 5'-position of 6-O-methyl-norlaudanosoline. 14 Using CH3-SAM as one Substrate and this novel enzyme as catalyst it was possible to synthesize 5'- 14 CH3-nororientaline with excellent yields. Using callus of A. platyceras, feeding experiments with the labelled Substrate showed that nororientaline was ex- Kompendium tensively metabolized by Argemone, yielding several alkaioids which are now being investigated. This finding demonstrates that the new enzyme is of phy- der siological importance in the plant tissue. Our results confirm previous findings [1, 2] that 6- Phytotherapie O-methylation clearly precedes methylation at C-7 and methylation at ring C within the reticuline/orien• Von S. CHRUBASIK und J. CHRUBASIK, Freiburg taline pathway. In addition the results obtained on the specific 1983. 156 Seiten, 15,5x23 cm, kartoniert NLS-6-O-methyltransferase [2] and especially the 5'- DM 34,— ISBN 3-7773-0618-5 O-methyltransferase described here clearly demon- strate that O-methylation precedes N-methylation. Der Trend zur Naturheilkunde und da besonders zur Phytotherapie nimmt zu. Doch um die Heil• kräuter therapeutisch nutzen zu können, bedarf Acknowledgements es eines intensiven Studiums. Dieses Kompendium soll Studierenden und Ärzten helfen, die interne Our thanks are due to Miss GABRIELE WEBER for excellent tech- Applikation pflanzlicher Drogen zu erleichtern und nical assistance and Dr. H. IBELGAUFTS for his kind linguistic help. dadurch zur Gesunderhaltung wie zur Behandlung This investigation was supported by SFB 145 of Deutsche For• von Krankheiten beizutragen. Das Buch ist für schungsgemeinschaft, Bonn, including a Finnigan MAT 44S in- alle Ärzte, die sich für die Phytotherapie inter• strument and by Fonds der Chemischen Industrie (literature Provi• essieren oder diese anwenden. sion).

References Ich bestelle aus dem Hippokrates Verlag Stuttgart durch die Buchhandlung: (1) Brochmann-Hanssen, E., C- (4) Batterby, A. R. in Taylor, W. H. Chen, R. Chen, H.-C. Chiang, I. and A. R. Battersby (eds.): Oxi- Expl. CHRUBASIK/CHRUBASIK, Kompendium A. Leung and K. McMurtrey: J. dative coupling of phenols, p. 119, der Phytotherapie. DM 34 — Chem. Soc. Perkin 1,1975, 1531. New York 1967, Marcel Dekker. ISBN 3-7773-0618-5 (2) Rueffer, M., N. Nagakura and (5) Tsang, Y.-F. and R. K. Ibra• M. H. Zenk: Planta Med., in press him: Z. Naturforsch. 34c, 46 (1983). (1979). (3) Brochmann-Hanssen, E., C- (6) Poulton, J. E., Stumpf, P. K. Unterschrift H. Chen, H.-C. Chiang, C.-C. Fu and E. E. Conn (eds.): The Bioche- and H. Nemoto: J. Pharm. Sei. 62, mistry of Plants, Vol. 7, p. 667, 1291 (1973). New York 1981, Academic Press. Name (möglichst Stempel)

Straße Address: Pharmazeutische Biologie, Karlstraße 29, Ort D-8000 München 2, Federal Republik ofGermany

Datum (Preisänderung vorbehalten) Hippokrates Verlag Stuttgart 57