Journal of Medicinal Plant Research

Planta medica

Organ der Gesellschaft für Arzneipflanzen­forschung

Editor-in-Chief E . Reinhard, Tübingen Pharmazeutisches Institut Auf der Morgenstelle 8 D-7400 Tübingen

Editorial Board H . P. T. Ammon, Tübingen W. Barz, Münster E . Reinhard, Tübingen

•O. Sticher, Zürich H . Wagner, München M . H . Zenk, München

Advisory Board N . Anand, Lucknow R. Anton, Strasbourg

A . Baerheim-Svendsen, Leiden

H . Böhm, Halle F. Bohlmann, Berlin A . Cave, Chatenay-Malabry P. Delaveau, Paris Ding Guang-sheng,

Shanghai C. -J. Estler, Erlangen N . Farnsworth, Chicago H . Floss, Columbus H . Friedrich, Münster D . Fritz, Weihenstephan A . G . Gonzalez, La Laguna O. R. Gottlieb, Sao Paulo E . Graf, Köln H . Haas, Mannheim

E . Hecker, Heidelberg R. Hegnauer, Leiden W. Herz, Tallahassee K . Hostettmann, Lausanne H . Inouye, Kyoto M . A . Iyengar, Manipal F. Kaiser, Mannheim F. H . Kemper, Münster W. R. Kukovetz, Graz J. Lemli, Leuven Liang Xiao-tian, Beijing M . Lounasmaa, Helsinki M . Luckner, Halle J. Lutomski, Poznan H . Menßen, Köln E . Noack, Düsseldorf J. D . Phillipson, London

J. M . Rowson, Mablethorpe M . v. Schantz, Helsinki K . F. Sewing, Hannover E . J . Shellard, London S. Shibata, Tokyo Ch. Tamm, Basel W. S. Woo, Seoul Xiao Pei-gen, Beijing

Contents

Volume 49,1983

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

II

Contents 49,1983

A t t a - u r - R a h m a n , B a s h i r , M . \ Isolation of New Alkaloids from C a t h a r a n t h u s roseus 124

A t t a - u r - R a h m a n , N i s a , M . , F a r h i , S.: Isolation of Moenjod-aramine from Buxus p a p i l o s a 126

B a l s e v i c h , J . , K u r z , W. G. W.: The Role of 9- and/or 10-oxygenated Derivatives of Geraniol, Geranial, Nerol, and Neral in the Biosynthesis of Loganin and Ajmalicine 79

Bassleer, R., M a r n e t t e , J . - M . , W i l i q u e t , P h . , D e P a u w - G i l l e t , M . - C L , C a p r a s s e , M . , A n g e n o t , L . \ Etüde complemen-taire de la cytotoxicite de la melinonine F, alcaloide derive de la ß-carboline (Complementary Study of Cytotoxic Activity of Melinonine F) 158

Becker, H . , C h a v a d e j , S., W e b e r l i n g , F . : Valepotriates in V a l e r i a n a t h a l i c t r o i d e s 64

Becker, H . , H e r o l d , S.: RP-8 als Hilfsphase zur Akkumula­tion von Valepotriaten aus Zellsuspensionskulturen von V a l e r i a n a w a l l i c h i i (RP-8 Auxiliary Phase for the Accu-mulation of Valepotriates from Cell-Suspension-Culture of V a l e r i a n a w a l l i c h i i ) 191

B o u n t h a n h , C, R i c h e r t , L . , Beck, J . P . , H a a g - B e r r u r i e r , M . , A n t o n , R.: The Action of Valepotriates on the Synthesis of D N A and Proteins ofCultured Hepatoma Cells. . . . 138

B r i a n q o n - S c h e i d , F . , L o b s t e i n - G u t h , A . , A n t o n , R.: H P L C Separation and Quantitative Determination of Biflavones in Leaves from G i n k g o b i l o b a 204

C a p u t o , O,, D e l p r i n o , L . , V i o l a , F . , C a r a m i e l l o , R., B a l -l i a n o , G.: Biosynthesis of Sterols and Triterpenoids in Tissue Cultures of C u c u r b i t a m a x i m a 176

C h a g n o n , M . , N d i b w a m i , A . , D u b e , S., B u m a y a , A r . Ac-tivite Anti-Inflammatoire d'Extraits de C r a s s o c e p h a l u m m u l t i c o r y m b o s u m (Anti-inflammatory Action of an Ex-tract from C r a s s o c e p h a l u m m u l t i c o r y m b o s u m ) 255

C h a t t o p a d h y a y , S., C h a t t o p a d h y a y , U., M a t h u r , P . P . , S a i n i , K . S., G h o s a l , S.: Effects of Hippadine, an Amarylli-daceae Alkaloid, on Testicular Function in Rats 252

Chen W e i m i n g , Yan Y a p i n g , L i a n g X i a o t i a n : Alkaloids from Roots of A I s t o n i a yunnanensis 62

Dehaussy, H . , T i t s , M . , A n g e n o t , L . \ La guattegaumerine, nouvel alcaloide bisbenzylisoquinoleinique de G u a t t e r i a g a u m e r i (Guattegaumerine, New Bisbenzylisoquinoline Alkaloid from G u a t t e r i a g a u m e r i ) 25

D o m i n g u e z , X . A . , F r a n c o , R., C a n o , G., M a . Consue l o G a r c i a , F . , X o r g e A , D o m i n g u e z , S. J r . , L e o n a r d o de la P e n a , M . : Isolation of a New Furano-l,4-Naphthaquinone, Diodantunezone from L a n t h a n a a c h y r a n t h i f o l i a . . . . 63

E n d o , K . , O s h i m a , Y., K i k u c h i , H . , K o s h i h a r a , Y.} H i k i n o , H . : Hypotensive Principles of U n c a r i a H o o k s 188

E n g e l s h o w e , R.: Dimere Proanthocyanidine als Gerbstoff­vorstufen in J u n i p e r u s c o m m u n i s (Tannin Producing DimericProanthocyanidins in J u n i p e r u s c o m m u n i s ) . . . 170

E s t e r l , A . , G a b , S., B i e n i e k , D . \ Zur Kenntnis der Inhalts­stoffe von I s e r t i a h y p o l e u c a (On The Constituents of I s e r t i a h y p o l e u c a ) 244

F o u r n i e r , G., P a r i s , M . : Mise en Evidence de Cannabinoides Chez P h e l i p a e a r a m o s a , Orobanchacees, Parasitant le Chanvre, C a n n a b i s s a t i v a , Cannabinacees (Detection of

Cannabinoi'ds in P h e l i p a e a r a m o s a , a Parasite of C a n n a -b i s s a t i v a ) 250

G a l u n , £., A v i v , D. , D a n t e s , A . y F r e e m a n , A . \ Biotransfor­mation by Plant Cells Immobilized in Cross-Linked Poly-acrylamide-Hydrazide. Monoterpene Reduction by En-trapped M e n t h a C e l l s 9

G h o s a l , Sh., S i n g h , A . K . , B i s w a s , K . : New6-Aryl-2-pyrones from G e n t i a n a p e d i c e l l a t a 240

H a f e z , A . y A d o l f , W., H e c k e r , E . \ Active Principles of the Thymelaeaceae. III. Skin Irritant and Cocarcinogenic Factors from P i m e l e a s i m p l e x 3

l e v e n , M . , v a n den B e r g h e , D . A . , V l i e t i n c k , A . J . : Plant Antiviral Agents. IV. Influence of Lycorine on Growth Pattern of Three Animal Viruses 109

I s h i g u r o , K . , Y a m a k i , A/., T a k a g i , 5.: Studies on Iridoid-related Compounds; III: Gentiopicral, the Aglucone of Gentiopicroside 208

J a k o v l e v , V., I s a a c , O., F l a s k a m p , E . : Pharmakologische Untersuchungen von Kamillen-Inhaltsstoffen, VI . Unter­suchungen zur antiphlogistischen Wirkung von Chama-zulen und Matricin (Pharmacological Investigations with Compounds of Chamomile, V I . Investigations on the Antiphlogistic Effects of Chamazulene and Matricine) 67

J o s h i , K . C , S i n g h , P . , T a n e j a , S.: A Sesquiterpenoid Naph-thol from K y d i a c a l y c i n a 127

Jossang, A . , Leboeuf, M . , C a b a l i o n , P . , C a v e , A . : Alcaloides des Annonacees. X L V : Alcaloides de P o l y a l t h i a n i t i -d i s s i m a (Alkaloids from Annonaceae. X L V : Alkaloids of P o l y a l t h i a n i t i d i s s i m a ) 20

K i m u r a , Y., O h m i n a m i , H . , O k u d a , H . , B a b a , K . , K o z a w a , M . , A r i c h i , S.: Effects of Stilbene Components of Roots of P o l y g o n u m ssp. on Liver Injury in Peroxidized Oil-fed Rats 51

K i s i e l , W.: 8-Epidesacylcynaropicrin from C r e p i s c a p i l l a r i s 246 K i s o , Y., Suzuki, Y., W a t a n a b e , N . , O s h i m a , Y., H i k i n o , H . :

Antihepatotoxic Principles of C u r c u m a l o n g a Rhizomes 185 K i s o , Y., T o h k i n , M . , H i k i n o , H . \ Assay Method for Anti­

hepatotoxic Activity Using Carbon Tetrachloride In-duced Cytotoxicity in Primary Cultured Hepatocytes . . 222

K r a m , G.f F r a n z , G.: Untersuchungen über die Schleim-polysaccharide aus Lindenblüten (Analysis of Linden Flower Mucilage) 149

Krüger, £>., J u n i o r , P . , W i c h t l , M . \ Neue Cardenolidglyko-side aus D i g i t a l i s l a n a t a (New Cardiac Glycosides from D i g i t a l i s l a n a t a ) 74

L a n g h a m m e r , L . , Schulze, G., G u j e r , R., M a g n o l a t o , D . , H o r i s b e r g e r , M . \ Isolation and Structure of a Rarely Occurring Cyanidanol Glycoside from Cortex Betulae . 181

L e m l i , J . , C u v e e l e , /., V e r h a e r e n , E . \ Chemical Identification of Alexandrian and Tinnevelly Senna. Studies in the Field of Drugs Containing Anthracene Derivatives X X X I V 36

L o n g - Z e L i n , W a g n e r , H . t S e l i g m a n n , ö.\ Thalifaberine, Thalifabine and Huangshanine, Three New Dimeric Aporphine-Benzylisoquinoline Alkaloids 55

Contents III

M a n - P o W o n g , T e h - C h a n g C h i a n g , H s o n - M o u C h a n g : Chem­ical Studies on Dangshen, the Root of C o d o n o p s i s p i l o -s u l a 60

M i s a w a , M . , H a y a s h i , M . , T a k a y a m a , S.: Production of Antineoplastic Agents by Plant Tissue Cultures. I. In-duction of Callus Tissues and Detection of the Agents in Cultured Cells 115

N a h r s t e d t , A . , W r a y , V., G r o t J a h n , L . , F i k e n s c h e r , L . H . , H e g n a u e r , R.: New Acylated Cyanogenic Diglycosides from Fruits of A n t h e m i s c a i r i c a 143

N o m u r a , T., F u k a i , T.} S h i m a d a , T., I h - S h e n g C h e n : Com-ponents of Root Bark of Morus australis. I. Structure of a New 2-Arylbenzofuran Derivative, Mulberrofuran D . 90

O b a t a - S a s a m o t o , H . , K o m a m i n e , A . : Effect of Culture Con-ditions on D O P A Accumulation in a Callus Culture of S t i z o l o b i u m hassjoo 120

O h i r i , F. C, V e r p o o r t e , R., B a e r h e i m Svendsen, A . : ! H N M R Chemical Shift Values for Aromatic Protons in 2,3,9,10-and 2,3,10,11-tetrasubstituted Tetrahydroprotoberberine Alkaloids 162

O h i r i , F . C , V e r p o o r t e , R., B a e r h e i m Svendsen, /i.:Tertiary Phenolic Alkaloids from C h a s m a n t h e r a dependens . . . 17

O j e w o l e , J . A . O., A d e s i n a , S. K . : Mechanism of the Hypo-tensive Effect of Scopoletin Isolated from the Fruit of T e t r a p l e u r a t e t r a p t e r a 46

O j e w o l e , J . A . O., A d e s i n a , S. K . : Cardiovascular and Neuro-muscular Actions of Scopoletin from Fruit of T e t r a p l e u r a t e t r a p t e r a 99

O k o g u n , J . L , Adeboye, J . O., O k o r i e , D . A . \ Novel Struc-tures of two Chromone Alkaloids from Root-Bark of S c h u m a n n i o p h y t o n m a g n i f i c u m 95

P a l a c i o s , P . , G u t k i n d , G., R o n d i n a , R. V. D . , de T o r r e s , R., C o u s s i o , J . D . : Genus B a c c h a r i s . II. Antimicrobial Activ­ity of B . c r i s p a and B . n o t o s e r g i l a 128

Pandey, V. B . , S i n g h , J . P . , M a t t o c k s , A . R., B a i l e y , E . \ A Note on "Isolation and Pharmacological Action of Helio-trine, the Major Alkaloid of H e l i o t r o p i u m i n d i c u m Seeds" 254

P a t h a k , V. P . , S a i n i , T. R., K h a n n a , R. N . : A New Furano-flavone from Seeds of P o n g a m i a g l a b r a 61

P e r e r a , P . , S a n d b e r g , F . , v a n Beek, T. A . , V e r p o o r t e , R.: Tertiary Indole Alkaloids of T a b e r n a e m o n t a n a d i c h o -t o m a Seeds 28

P e r e r a , P . , v a n Beek, T. A . , V e r p o o r t e , R.\ Dichomine, a Novel Type of Iboga Alkaloid 232

R a v i d , U., P u t i e v s k y , E . : Constituents of Essential Oilsfrom M a j o r a n a s y r i a c a , C o r i d o t h y m u s c a p i t a t u s and S a t u r e j a t h y m b r a 248

Röder, E . , W i e d e n f e l d , H . , H o n i g , A . : Pyrrolizidinalkaloide aus Senecio a u r e u s 57

Rueffer, M . , N a g a k u r a , N . , Zenk, M . H : A Highly Specific O-Methyltransferase for Nororientaline Synthesis Isolated from A r g e m o n e p l a t y c e r a s Cell Cultures 196

Rueffer, M . , N a g a k u r a , N . , Zenk, M . H . : Partial Purification and Properties of S-Adenosylmethionine: (R), (S)-Nor-laudanosoline-6-O-Methyltransferase from A r g e m o n e p l a t y c e r a s Cell Cultures 131

S a r i y a r , G . : Alkaloids from P a p a v e r t r i n i i f o l i u m of Turkish Origin 43

Seip, E . H . , O t t , H . H . , H e c k e r , E . : Skin Irritant and Tumor Promoting Diterpene Esters of the Tigliane Type from the Chinese Tallow Tree ( S a p i u m s e b i f e r u m ) 199

Sepulveda-Boza, S., F r i e d r i c h s , £., Puff, H . , B r e i t m a i e r , E . \ Ein iso-Homoprotoberberin-Alkaloid aus den Wurzeln von B e r b e r i s a c t i n a c a n t h a (An iso-Homoprotoberberin-Alkaloid from the Roots of B e r b e r i s a c t i n a c a n t h a ) . . . . 32

Shoyama, Y., H a t a n o , K . , N i s h i o k a , I . : Clonal Multiplica-tion of P i n e l l i a t e r n a t a by Tissue Culture 14

S t o i a n o v a - I v a n o v a , B . , B u d z i k i e w i c z , H . , K o u m a n o v a , B . , T s o u t s o u l o v a , A . , M l a d e n o v a , K . , B r a u n e r , A . : Essential Oil of C h r y s a n t h e m u m i n d i c u m 236

T e h - C h a n g C h i a n g , H s o n - M o u C h a n g , M a k , T h . C. W.: New Oleanene-type Triterpenes from A b r u s p r e c a t o r i u s and X-ray Crystal Structure of Abrusgenic Acid-Methanol 1:1 Solvate 165

v a n der S l u i s , W. G., v a n der N a t , J . M . , Spek, A . L . , I k e -s h i r o , Y., L a b a d i e , R. P . : Gentiogenal, a Conversion Pro-duct of Gentiopicrin (Gentiopicroside) 211

W a g n e r , H . , S c h w a r t i n g , G., V a r l j e n , B a u e r , R., H a m -d a r d , M . E . , E l - F a e r , M . Z., B e a l , J . \ Die chemische Zu­sammensetzung der Convolvulaceen Harze IV. Die Gly-kosidsäuren von I p o m o e a q u a m o c l i t , I . l a c u n o s a , I . p a n -d u r a t a und C o n v o l v u l u s a l - s i r e n s i s (Chemical Consti­tuents of the Convolvulaceae-Resins IV. The Glycosidic Acids of I p o m o e a q u a m o c l i t , I . l a c u n o s a , 1. p a n d u r a t a and C o n v o l v u l u s a l - s i r e n s i s ) 154

W a t a n a b e , K , W a t a n a b e , H . , G o t o , Y., Y a m a g u c h i , M . , Y a m a m o t o , N . , H a g i n o , K . \ Pharmacological Properties of Magnolol and Hönokiol Extracted from M a g n o l i a offi-c i n a l i s : Central Depressant Effects 103

W i l l u h n , G., Röttger, P . - M . , M a t t h i e s s e n , U.: Helenalin-und 11,13-Dihydrohelenalinester aus Blüten von A r n i c a m o n t a n a (Helenalin- and 11,13-Dihydrohelenalinester from Flowers of A r n i c a m o n t a n a ) 226

W i t t e , L . , B e r l i n , J . , W r a y , V., Schubert, W., K o h l , W., Höfle, G. , H a m m e r , J . : Mono- and Diterpenes from Cell Cultures of T h u j a o c c i d e n t a l i s 216

Y a n g M i n g h e , Chen Y a n y o n g : Steroidal Sapogenins in D i o s -c o r e a c o l l e t t i i 38

Yu D e - Q u a n , D a s , B . C.: Structure of Hydroxymuscopyri-dine A and Hydroxymuscopyridine B , Two New Constit­uents ofMusk 183

Yu D e - q u a n , Y., D a s , B . C.: Alkaloids of A c o n i t u m b a r b a t u m 85

IV

Subject Index 49,1983

A

Abietane 220 Abrusgenic acid 165 Abruslactone 165 Acevaltrate 64, 191 Adaptogenes 240 Alkaloids 17, 20, 25, 28, 32, 43, 55, 57, 62,

79, 85, 95,109,124, 126,131,158,162, 188,196, 232, 244, 252, 254

Amyrin 177 Antineoplastic agents 115 Antiviral agents 109 Apigenin 128 Aporphine alkaloids 20 Aporphine derivatives 43 Apparicine 232 Arabinose 149 Arylbenzofuran derivatives 90 Avenasterol 177

B

Benzylisoquinoline alkaloids 131 Berberine alkaloids 32 Biflavones 204 Bilobetin 204 Biosynthesis of alkaloids 131, 196 Biosynthesis of ajmalicine 79 Biosynthesis of loganin 79 Biosynthesis of sterols 176 Biosynthesis of triterpenoids 176 Biotransformation 9 Bisabolol 67 Bisbenzylisoquinoline alkaloids 20, 25, 55 Bisnorargemonine 14 Borneol 236 Bornyl acetate 236

C

Cadambine 188 Caffeic acid 186 Camphor 216 Cannabinoi'ds 250 Cardenolides 74 Carvacrol 216 Catechin 181 Catechinxylopyranoside 181 Cell cultures 9, 14 79, 115, 120, 131, 176,

191,196,216 Chamazulene 67 Cholestanol 177 Chromone alkaloids 95 Chrysanthenone 236 Chrysosplenetin 63 Cinchonamine 244 Cinnamic acid 186 Clonal multiplication 14

Cocarcinogens 3 Convolvulaceae resins 154 Coreximine 14 Coronaridine 28 Coumaric acid 186 Coumarins 46, 99 Cucurbitacin 176 Curcumin 185 Curcuminoids 185 Cyanogenic glycosides 143 Cycloartenol 177 Cymenol 216 Cynarin 224 Cynaropicrin 246 Cytotoxic action 158

Gentiogenal 211 Gentiopicral 208 Gentiopicrin 211 Gentiopicroside 208, 211 Geranial 79 Geraniol 79 Ginkgetin 204 Glycosidic acids 154 Glycyrrhetinic acid 224 Glycyrrhizin 224 Govanine 17 Guajazulene 67 Guattegaumerine 25

H

D

Daphnane derivatives 3 Dauricine 20 Daurisoline 20 Dehydrohelenalinester 226 Deoxyphorbol 199 Dichomine 232 Didrovaltrate 64, 138, 191 Diginatigenin derivatives 74 Digoxigenin derivatives 74 Dihydrocadambine 188 Dihydroquinamine 244 Diodantunezone 63 Diosgenin 38 Diterpene esters 3, 199 Diterpenes 216 Diterpenoid alkaloids 85 Dopa 120

Enzymes 196 Epilumicin 143 Epoxyphorbol 199 Erythrocentaurin 212 Essential oils 9, 67, 216, 236, 248

Fenchone 220 Ferruginol 220 Ferulic acid 186 Flavonoids 61, 63, 90, 128, 204 Fluorocarpamine 124 Fridelin 60 Furoflavones 61

Helenalin 226 Helenalinester 226 Helenanolides 226 Heliotrine 254 Hepatoma cells 138 Hibiscone 127 Hibiscoquinone 127 Hinikione 220 Hinokiol 220 Hippadine 252 Homovaltrate 64, 191 Honokiol 103 Huangshanine 55 Hydroxymuscopyridine 183 Hydroxymusizin glycoside 36

Iboga alkaloids 232 Ibogamine 28 Iboxygaine 232 Immobilized cells 9 Indicine 254 Indole alkaloid glucoside 188 Indole alkaloids 28, 62, 79, 124. 158, 188,

232 Iridoids 208,211 Isochinoline alkaloids 196 Isodihydrocadambine 188 Isoginkgetin 204 Isoquinoline alkaloids 32 Isothujone 220 Isovaltrate 64, 191

K

Kuwanon 90

G

Galactose 149 Genkwanin 128

L

Lasiocarpine 254 Laudanosoline 196

Subject Index V

Levomenol 67 Liliolide 240 Lindolhamine 20 Linoleic acid 220 Liriodenine 20 Lumicin 143 Lycorine 109

M

Magnolol 103 Mandelonitril glycosides 143 Matricine 67 Melionine 158 Mentenediol 220 Menthone 9 Methionine 224 Methylabrusgenat 165 Methylnorlaudanosoline 196 Methyltransferase 131, 196 Moenjodaramine 126 Monoterpenes 216 Morphinane derivatives 43 Morusin 90 Mucilage 149 Myrtanol 216

N

Naphthalene glycosides 36 Naphthaquinones 63 Neolignane derivatives 103 Neomenthol 9 Neral 79 Nerol 79 Norlaudanosoline 131 Normacusine 62 Nororientaline 196 Norreticuline 131

O

Opium alkaloids 131 Orientaline 196

P

Pallidine 14 Palmatic acid 220 Papaver alkaloids 43 Pedicellatin 240

Pedicellin 240 Penduletin 63 Perakine 62 Perivine 232 Phorbol 199 Phytosterols 60, 176 Piceid 51 Picroside 224 Pimelea factors 3 Pleiocarpamine 124 Podophyllotoxin 224 Polysaccharides 149 Proanthocyanidins 170 Protoberberin derivatives 32 Protoberberine alkaloids 20 Protocatechuic acid 186 Pseudoguaianolides 226, 246 Puberaconitidine 85 Puberaconitine 85 Puberanidine 85 Puberanine 85 Pulegone 9 Pyrrolizidine alkaloids 57

Resins 154 Resveratrol 51 Reticuline 20, 131 Rhamnose 149 Rhoeadine derivatives 43

Steroidal sapogenins 38 Stigmastenol 177 Stilbene51 Sugiol 220 Swertiamarin 240

Tabersonine 28 Tacaman alkaloids 232 Tannins 181 Taraxerol 60 Terpineol 216 Tertiary phenolic alkaloids 14 Tetrahydroalstonine 62 Tetrahydroisochinoline alkaloids 196 Tetrahydroprotoberberine alkaloids 162 Thalifaberine 55 Thalifabine 55 Thujaplicin derivatives 216 Thujone 220 Tigliane derivatives 3 Tinnevellin glycoside 36 Tripdiolide 109 Triterpenoids 60, 165 Tropolone 220 Tumor promotors 199

U

Ursolic acid 240 Ushinsunine 20

S

Sabinene 220 Sapogenins 38 Saponins 38, 60 Sarpagine 62 Schumannificine 95 Sciadopitysin 204 Scopoletin 46, 99 Secoiridoids 208,211 Sesquiterpene lactones 226,246 Sesquiterpenes 127 Silybin 224 Simplexin 3 Sinapic acid 186 Sitosterol 220 Skin irritants 199 Spinasterol 177 Stemmadenine 28 Stepholidine 20 Steroidal alkaloids 126

V

Valepotriates 64, 138, 191 Valtrate 64, 138,191 Vellosimine 62 Vindolinine derivatives 124 Vinorine 62 Voacangine 28 Voacristine 232 Voaphylline 28,232 Vobasine 232

Y

Yamogenin 38

Z

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 C a r y o p h y l l a c e a e Apiaceae sub U m b e l l i f e r a e Arecaceae sub P a l m a e Asteraceae sub C o m p o s i t a e Brassicaceae sub Cruciferae Clusiaceae sub G u t t i f e r a e Fabaceae sub L e g u m i n o s a e (including M i -

mosoideae = M i m o s a c e a e , Caesalpini-oideae = C a e s a l p i n i a c e a e and Papilion-oideae = P a p i l i o n a c e a e = Fabaceae sensu stricto)

Hypericaceae sub G u t t i f e r a e Lamiaceae sub L a b i a t a e Oenotheraceae sub O n a g r a c e a e Poaceae sub G r a m i n e a e

Abrus cantoniensis 165 Abrus precatorius 165 Aconitum barbatum 85 Adlumia fungosa 133 Albertisia papuana 22 Alstonia yunnanensis 62 Althaea officinalis 152 Amaryllidaceae 109, 252 Annonaceae 20, 25 Anthemis altissima 143 Anthemis cairica 143 Anthocephalus cadamba 189 Apocynaceae 28, 62, 79,124, 232 Araceae 14 Argemone intermedia 133 Argemone platyceras 131, 196 Arnica montana 226 Artemisia caruthii 67

B

Baccharis crispa 128 Baccharis megapotamica 116 Baccharis notosergila 128 Baliospermum montanum 200 Berberidaceae 32,133 Berberis actinacantha 32 Berberis henryana 133 Berberis stolonifera 133 Berberis wilsonae 133 Betula species 181 Betulaceae 181 Blackstonia perfoliata 211 Bombax malabaricum 127 Boraginaceae 254 Brucea antidysenterica 116 Bryonia dioica 176 Buxaceae 126 Buxus papulosa 126

Cacalia floridana 57 Caesalpinia gilliesii 116 Caesalpiniaceae 36 Campanulaceae 60 Cannabis sativa 250 Cassia angustifolia 36 Cassia senna 36 Catharanthus roseus 79,124 Celastraceae 115 Cephalotaxaceae 115 Cephalotaxus harringtonia 115 Chamomilla recutita 67 Chasmanthera dependens 17,162 Chelidonium majus 133 Chlorella ellipsoidea 178 Chrysanthemum indicum 236 Cissampelos mucronata 133 Clivia miniata 109 Codonopsis pilosula 60 Colchicum speciosum 116 Compositae 57,67,128,143,226,236,246,

255 Convolvulaceae 154 Convolvulus al-sirensis 154 Convolvulus microphyllus 154 Coridothymus capitatus 248 Corydalis pallida 133 Corydalis sempervirens 133 Crassocephalum multicorymbosum 255 Crepis capillaris 246 Crepis virens 246 Crinum asiaticum 252 Crinum augustum 252 Crinum latifolium 252 Crinum pratens 252 Cucumis sativus 176 Cucurbita maxima 176 Cucurbita pepo 177 Cucurbitaceae 176 Cupressaceae 170, 216 Curcuma longa 185

D

Dictiostelium discoideum 177 Digitalis lanata 74 Digitalis schischkinii 74 Dioscorea collettii 38 Dioscoreaceae 38

Fagara zanthoxyloides 116 Fumaria officinalis 133

Gentiana pedicellata 240 Gentianaceae 208, 211, 240 Ginkgo biloba 204 Ginkgoaceae 204 Glaucium flavum 133 Guatteria gaumeri 25

H

Erythrina lithosperma 22 Eschscholtzia tenuifolia 133 Euphorbiaceae 199

Heliotropium indicum 116,254 Heliotropium supinum 254 Hibiscus species 127 Holacantha emoryi 116 Hura crepitans 3

I

Ipomoea lacunosa 154 Ipomoea operculata 154 Ipomoea pandurata 154 Ipomoea quamoclit 154 Ipomoea turpethum 154 Isertia hypoleuca 244

Juniperus communis 170

Kydia calycina 127

Labiatae 9, 248 Lantana achyranthifolia 63 Leguminosae 36, 46, 61, 99, 120,165 Lindera oldhamii 22, 26 Loganiaceae 158

M

Magnolia obovata 103 Magnolia officinalis 103 Magnoliaceae 103 Majorana syriaca 248 Malvaceae 127

Index of Names of Organisms Vl l

Maytenus bucchananii 116 Menispermaceae 17, 133, 162 Menispermum dauricum 22 Mentha species 9 Mimosaceae 46, 99 Moraceae 90 Morus alba 90 Morus australis 90 Moschus moschiferus .183 Mycobacterium species 10

N

Nicotiana tabacum 250

O

Ochrosia moorei 116 Origanum compactum 249 Origanum floribundum 249 Origanum hirtum 249 Origanum majorana 249 Origanum maru 248 Origanum smyrnaeum 249 Origanum syriacum 248 Orobanchaceae 250

P

Panax ginseng 60 Panax pseudo-ginseng 60 Papaver armeniacum 43 Papaver cylindricum 43 Papaver fugax 43 Papaver Orientale 196 Papaver somniferum 131,196 Papaver tauricola 43 Papaver triniifolium 43 Papaveraceae 43,131,196 Papilionaceae 120

Penicillium expansum 211 Phelipaea ramosa 250 Pimelea linifolia 7 Pimelea prostrata 3 Pimelea simplex 3 Pinellia ternata 14 Podophyllum hexandrum 223 Polyalthia beccarii 20 Polyalthia emarginata 20 Polyalthia nitidissima 20 Polyalthia oligosperma 20 Polyalthia oliveri 20 Polyalthia suaveolens 20 Polygonaceae 51 Polygonum cuspidatum 51 Polygonum multiflorum 51 Polypodium vulgare 181 Pongamia glabra 61 Putterlickia verrucosa 115

R

Ranunculaceae 55, 85,133 Rhus coriaria 248 Rubiaceae 95,188,244

S

Sapium sebiferum 199 Satureja capitata 248 Satureja peltieri 249 Satureja thymbra 248 Schistosoma japonicum 199 Schumanniophyton magnificum 95 Schumanniophyton problematicum 97 Scolytus multistriatus 181 Scolytus ratzeburgi 181 Scrophulariaceae 74 Senecio aureus 57 Stizolobium hassjoo 120 Streptomyces clavuligerus 10

Strychnos melinoniana 158 Strychnos usambarensis 158

T

Tabernaemontana dichotoma 28, 232 Tabernaemontana eglandulosa 232 Taxus brevifolia 116 Tetrapleura tetraptera 46, 99 Thalictrum dasycarpum 116 Thalictrum faberi 55 Thalictrum sparsiflorum 133 Thalictrum tuberosum 133 Thuja occidentalis 216 Thymelaeaceae 3 Thymus capitatus 248 Tilia species 149 Tiliaceae 149 Tripterygium wilfordii 115

U

Ulmus americana 181 Uncaria sinensis 188 Uncaria species 188

V

Valeriana edulis ssp. procera 64 Valeriana kilimandscharica 64 Valeriana thalictroides 64 Valeriana wallichii 138,191 Valerianaceae 64, 138, 191 Verbenaceae 63

Z

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

J o u r n a l o f Planta MoHirinal •••"IUI

1 9 8 3 , V o l . 4 9 , p p . 1 9 6 - 1 9 8 , © H i p p o k r a t e s V e r l a g G m b H Plant Re^aSI m e d O

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

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

* Lehrstuhl für Pharmazeut ische Biologie der Universität München, Federal Republic of Germany

** Kobe Women 's College of Pharmacy, Kobe 658, Japan

Received: August 8 , 1 9 8 3

Key Word Index:

Argemone platyceras] Papaveraceae; Cell Suspen­sion cultures; Alkaloid biosynthesis; 6-O-methyl-norlaudanosoline)-5' -O-methyltransferase.

Abstract

A highly specific new enzyme, S-adenosyl-L-me-thionine: (6-O-methyl-norlaudanosoline)-5; -O-me-thyltransferase which catalyses the formation of nor­orientaline from 6-O-methyl-norlaudanosoline and SAM was discovered, partially purified, and charac-terized. Argemone platyceras cell Suspension cultures served as enzyme source.

Introduction

Tetrahydroisoquinoline biosynthesis in plants in-volves a number of O- and N-methylation Steps [e.g. 1] of norlaudanosoline, the first precursor in the pathway. In a previous communication, we have des­cribed the isolation of a first specific O-methyltrans-ferase. In Argemone platyceras cell Suspension cultu­res this enzyme methylates predominantly the 6-0-position of (S)- as well as (R)-norlaudanosoline, me-diating the transfer of a methyl-group from S-adeno-syl-L-methionine to NLS [2]. Düring the purification of this enzyme, a second enzymatic activity was dis­covered which utilises 6-O-methyl-NLS to introduce a second methyl group in the 5;-position of the mole-cule to yield nororientaline. Again SAM serves as a methyl group donator. Nororientaline is a principal immediate precursor of papaverine in Papaver som­niferum [1] and is also undoubtedly converted to orientaline by N-methylation. Orientaline itself has

been isolated from opium poppy [3] and is metaboli-zed in Papaver Orientale via orientalinone to isothe-baine [4]. Therefore nororientaline can be consid-ered as a specific precursor of a number of isoquinoli-ne alkaloids in higher plants. This report describes the partial purification and characterization of a new enzyme which was designated S-adenosyl-L-methio-nine: (6-0-methyl-norlaudanosoline)-5'-0-methyl-transferase.

Material and Methods A r g e m o n e p l a t y c e r a s cells were cultivated as described pre­

viously [2]. Chemicals used were essentially those described in the preceeding communication [2]. The new enzyme, described here, was purified through Steps 1-4 as described previously [2]. How-ever, the enzyme was eiuted from hydroxyapatite columns at frac-tions 56-65 in contrast to the previously described NLS-6-O-me-thyltransferase which was eiuted between fractions 43 and 54. En­zyme assays were carried out as follows: (R, S)-6-0-methyl-nor-laudanosoline (0.3 mM), S A M - 3 H (10000 cpm, 0.1 mM), K P 0 4

2 -buffer, p H 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 200 ul Na 2C0 3-buffer (1 M , p H 9.5). The methylated product was extracted by adding 400 ul isoamylacohol and shaking for 45 min. After centrifugation for 5 min. in an Eppendorf centrifuge, 200 ul of the organic phase were transferred to scintillation vials and counted. A blank value of about 10 % (obtained by assay mixtures containing either no enzyme or no Substrate), had to be subtracted from all incubation mixtures. Recovery of thje methylated product was 92 % under these conditions.

The products were separated and identified by H P L C using a 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 time of the potential products were: norisoorientaline, 7.14 min.; nororientaline, 8.20 min.; norprotosinomenine, 8.84 min.; norre-ticuline, 10.94 min.

Preparative isoiations were done using the above incubation mixture x 100. The reaction product was extracted by ethylacetate and purified twice by thin layer chromatography (Si-gel; solvent Systems: 1) Chloroform : methanol : acetic acid : water = 18:6:3:0.3; 2) acetone : Chloroform : diethylamine = 5:4:1). Mass spectra were determined in a Finnigan M A T 44S instrument.

Results and Discussion

Abbreviations: NLS = Norlaudanosoline; S A M = S-Adeno-syl-L-methionine; S A H = S-Adenosylhomocysteine.

Düring the purification of the 6-O-methyltransfe-rase evidence was obtained that its reaction product

O-Methyltransferase for Nororientaline Synthesis 1 9 7

Table I Purification protocol for the ö ' -O-methyl t ransferase f rom 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

( N H 4 ) 2 S 0 4 precipitation 23 5733 10.02 24.9 100 1.4

GelTiltration 69 5703 0.22 375.5 99.48 20.9

DEAE chromatography 35 1127 0.07 506.5 19.65 28.2

Hydroxyapati te 12 327.7 0.038 718.8 5.7 40.0

Table II Some characterist ics of (6-0-methy lnor laudanoso l ine) -5 ' -0-

methyltransferase f rom A. platyceras cell cultures

Characterist ics 5 ' -0-methy l t ransferase

Molecular weight 47.000

pH-opt imum 7.5

Temperature Optimum 35° C

K M for 6-O-methylnor laudano-

soline 0.4 m M

K M for SAM 0.05 m M

Kj for SAH 0.034 m M

Table III Substrate specifity of 40-fold purified 5 ' -0-methy l t ransferase

using S A M as methyl group donator

Substrate pkat/mg % Product of

reaction

(R, S) -6-0-methy l -

norlaudanosol ine 314 100 Nororientaline

(R)-Norlaudanosol ine 0 0 -(S)-Norlaudanosol ine 0 0 -(R, S)-Laudanosol ine 0 0 -

is transformed by the introduction of a second methyl group in the presence of excess SAM and crude en­zyme preparations. The two enzymes involved are clearly separated by chromatography on hydroxya­patite. The enzyme was partially purified by using (R,S)-6-0-methyl-norlaudanosoline as Substrate. As shown in Table I the enzyme could be recovered with 6 % yield. Purification was about 40-fold.

The characteristic properties of the enzyme are li-sted in Table II. The enzyme has a half life of 33 hrs at 30° C and retains füll enzymatic activity in the pres­ence of 0.05 % NaN 3 at 4° C after 4 weeks. The en­zyme is, however, completely inactivated by freezing even in the presence of 30 % glycerol. The enzyme is inhibited completely by Fe 3 + and H g 2 + ions at 5 mM concentration and is strongly inhibited by 5 mM p-chloromercuribenzoate (84 %), N-ethylmaleimide (82 %), and 10mM jodobenzoic acid (79 %). This in-dicates that SH-groups are present at the active Cen­ter of the enzyme. Similar findings have been repor-

+ SAM

6-0-Methylnorlaudanosoline

ted for the unspecific meta and p a r a directing cate-chol-O-methyltransferases from plant origin [5]. The general properties of the enzyme, described here, are very similar to the previously described norlaudano-soline-6-O-methy ltransferase.

The Substrate specifity of the 5'-0-methyltransfe-rase was of considerable interest. 18 Substrates were tested for their ability to accept methyl groups from SAM catalysed by S-adenosyl-L-methionine: (6-0-methyl-norlaudanosoline)-5'-0-methyltransferase. None of the phenols, phenylpropanoids, flavonoids, coumarins, and biogenic amines tested, served as Substrates. Out of 12 different isoquinoline alkaloids tested [2] only (R,S)-6-0-methyl-norlaudanosoline was methylated. Surprisingly neither (R) or (S)-nor-laudanosoline nor (R, S)-laudanosoline served as a Substrate (Table III) which shows a hitherto unique Substrate specifity of an O-methyltransferase in plant kingdom [6]. Since the 6-O-methyl-norlaudanosoline used was a racemate, it is not known whether either

+ SAH

Nororientaline

1 9 8 Rueffer, Nagakura, Zenk

Hippokrates

Kompendium der Phytotherapie Von S. C H R U B A S I K und J. C H R U B A S I K , Freiburg

1983. 156 Seiten, 15,5x23 cm, kartoniert D M 34,— ISBN 3-7773-0618-5

Der Trend zur Naturheilkunde und da besonders zur Phytotherapie nimmt zu. Doch um die Heil­kräuter therapeutisch nutzen zu können, bedarf es eines intensiven Studiums. Dieses Kompendium soll Studierenden und Ärzten helfen, die interne Applikation pflanzlicher Drogen zu erleichtern und dadurch zur Gesunderhaltung wie zur Behandlung von Krankheiten beizutragen. Das Buch ist für alle Ärzte, die sich für die Phytotherapie inter­essieren oder diese anwenden.

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of the two stereoisomers is preferentially methyl­ated. As shown by HPLC (retention time 8.20 min.) 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.

Using 1 4 CH 3 -SAM as one Substrate and this novel enzyme as catalyst it was possible to synthesize 5'-14CH3-nororientaline with excellent yields. Using callus of A . platyceras, feeding experiments with the labelled Substrate showed that nororientaline was ex-tensively metabolized by Argemone, yielding several alkaioids which are now being investigated. This finding demonstrates that the new enzyme is of phy-siological importance in the plant tissue.

Our results confirm previous findings [1, 2] that 6-O-methylation clearly precedes methylation at C-7 and methylation at ring C within the reticuline/orien­taline pathway.

In addition the results obtained on the specific NLS-6-O-methyltransferase [2] and especially the 5'-O-methyltransferase described here clearly demon-strate that O-methylation precedes N-methylation.

Acknowledgements

Our thanks are due to Miss GABRIELE WEBER for excellent tech-nical assistance and Dr. H . IBELGAUFTS for his kind linguistic help. This investigation was supported by SFB 145 of Deutsche For­schungsgemeinschaft, Bonn, including a Finnigan M A T 44S in-strument and by Fonds der Chemischen Industrie (literature Provi­sion).

References

(1) Brochmann-Hanssen, E . , C -H. Chen, R. Chen, H.-C. Chiang, A. Leung and K. McMurtrey: J. Chem. Soc. Perkin 1,1975, 1531. (2) Rueffer, M . , N. Nagakura and M. H. Zenk: Planta Med., in press (1983). (3) Brochmann-Hanssen, E . , C -H. Chen, H.-C. Chiang, C.-C. Fu and H. Nemoto: J. Pharm. Sei. 62, 1291 (1973).

(4) Batterby, A. R. in Taylor, W. I. and A. R. Battersby (eds.): Oxi-dative coupling of phenols, p. 119, New York 1967, Marcel Dekker. (5) Tsang, Y.-F. and R. K. Ibra­him: Z. Naturforsch. 34c, 46 (1979). (6) Poulton, J. E . , Stumpf, P. K. and E. E. Conn (eds.): The Bioche-mistry of Plants, Vol. 7, p. 667, New York 1981, Academic Press.

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