UNIVERSITI PUTRA MALAYSIA

CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND THEIR BIOLOGICAL

ACTIVITIES

CHEW YEAN L I NG

FSAS 1995 7

CHEMI CAL CONST I TUENTS OF GLYCOSHIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND THE I R B I OLOGI CAL

ACTIVITIES

By

CHEW YEAN L I NG

The s is S ubmi t t ed in Fu l f i l me n t o f t he R e q u i r ements f or t h e De gree o f Ma s t e r o f S c i e nce

i n t he Fa c u l ty o f S c i ence a nd Env i r onme n t a l S t ud i e s , Unive r s i t i P e r t a nia n Ma l a ysia

June 19 9 5

ACKNOWLEDGEMENTS

I wou l d like t o ta k e this oppor tun i t y t o e x pr e s s

m y s i nce r e a pprec i a tlon and gra titude t o A s s o c . Prof .

Dr . Ha wa rdi Ra hma ni a nd Ass oc . P r o f . Dr . Sar i a h Meon

for t he inva l ua b l e advice and guida nce thr oughout the

c o u r se o f this pr oje ct, and Asso c . Pr o f . Dr . Mohd.

Aspo l l a h Hj. Suk a r i for his cons t a nt s uppor t .

I wou ld a l s o l ik e t o a c k nowl edge t he coope ra tion

and assistance o f the s ta f f o f the Chemi str y

De pa r tme n t o f UPM : En . Na z r i Ahmad , En . Z a i na l Abld i n

Kas i m , En. Ho hamad Abu S amah, En . Z a i na l Z a ha r i

Z a k a ria , En. Nordin I smail and En . Zainuddin Samad i .

La s t but n o t l e a s t , I wis h to tha n k P r o f . Norio

Aimi o f Chiba Un i ve r s i t y f or the run n i ng o f I H -NMR

and 13C-NHR s pe ctr a , Mr . S. Anth o ny Samy o f the

Bio l ogy De pa r t ment o f UPH a nd En . Ahmad Zainuddin

I br a h i m o f UK M f o r i d e n t i f ying a nd co l l ect i ng the

p lant mate ria l s , and En . K hir Rani a nd the sta f f o f

the Plant P r otecti o n Department o f UPM f or their help

a nd c o o pe r a t i o n d ur ing the bioas say s tudy .

i i

TABLE OF CONTENTS

ACKNOWLEDGEMENTS

L I S T OF TABLES

L I S T OF F I GURES

L I ST OF PLATES

L I ST OF ABBREV I AT I ONS

ABS TRACT

ABSTRAK

PART I EXTRACTI ON AND I S OLAT I ON OF CHEHI CAL COHPONENTS FROM GLYCOSHIS CALCICOLA AND G. RUPESTRIS ( RUTACEAE)

Page

11

v i

vii

x i

x

x i

xiii

I NTRODUCT I ON . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

L I TERATURE REVI EW. . ... . .. . . . . ... . . . ..... 3

OBJECTI VES OF S TUDy . . . . . . . . . . . . . . . . . . . . . 17

RESULTS AND D I S CUS S I ON . . . . . . . . . . . . . . . . . . 18

G l ycosm i s Cal c i co l a B . C . S t one . . . . . . . . . . . . . 1 8 Glycosm i s Calc i cola ( La ngk a v l I s l and ) ... 18 Glycosm i s Calclcola ( Temp ler P a r k ) . . . . . . 3 6

Glycosm i s R u pestr l s R i dley . . . . . . . . . . . . . . . . . 55 Glycosmi s R upes t r i s «S tem B a r k ) . . . . . . . . 5 5 Glycosm i s R upes tri s ( Lea ves ) . . . . . . . . . . . . 83

CONCLUS I ON . . • • . • • . • . • • . • • . • . . . . . . • . . . . . . . . . 8 4

i l l

EXPERIMENTAL............................... 85 Gene r a l M a t e r i a l s a nd Method s . . . . . . . . . . . . . . 85 Ext r ac t i o n o f Glycosmis Calclcola ( Langk a w i I s l a nd ) . . . . . . . . . . . . . . . . . . . . . . . . . . 8 7

I s o la t i o n o f GC1 (4 4 ) . . . . . . . . .. . . . . . . . . . . 87 I s o l a t i on o f S i t o s t e r o l (48)............ 89 Is o la t i on o f GC6 . . . . . . . . . . . . . . . . . . . . . . . . 9 0

Ext r ac t i on o f Glyco s m i n Ca l c i c o l a ( Te mp l e r Park ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 1

I s o l a t i on o f F l i nd e r s i n e (4 9)........... 9 1 I s o l a t i o n o f De s me t hoxyzanth o phyl l i n e ( 50 ) . . . . . . . . . . . 9 3

Ex t r a c t i on o f Glycosmis Rupestrls . . . . . . . . . . I s o l a t i on o f G l ycoma u r i n ( 2 2 ) . . . . . . . . . . . 9 4 I s o l a t i o n o f 7 -m e t h o xyg l ycoma u r i n (51)............... 9 6 I s o l a t i on o f De s -N-me t hyl a c r o nyc i ne ( 5 3 ) . . . . . . . . . . . . . 9 7 I s o l a t i on o f s t i gmas t - 7-e n o l (54 ).................... 9 8 I s o l a t i o n o f GCIS . . . . . . . . . . . . . . . . . . . . . . . 9 9

PART I I ANTI FUNGAL ASSAYS AGAINST CHAONEPHORA CUCURBITARIUH, COLLETOTRICHUH CAPSICI AND C. GLOEOSPORIOIDES

I NTRODUCT I ON . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102

OBJECT I VES OF S TUDy . . . . . . . . . . . . . . . . . . . . . . . . 1 0 6

LITERATURE REVIEW . . . . . . . . . . . . . . . . . . . . . . . . . . 107

MATER I AL S AND METHODS . . . . . . . . . . . . . . . . . . . . . . 1 1 1

P r e pa r a t i on o f P l a n t Extracts . . . . . . . . . . . . . . 1 1 1 Preparation of stock Cultures . . . . . . • . . . . . . . III Preparat i on o f Assay Media................. 1 1 2 Myce lia l Gr owth a nd Spor u l a t i on . . . . . . . . . . . . 1 1 2 S p o r e Ge r m i na t i on . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1 4 TLC B i oa s s a y . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1 5

RESULTS . • . • . • . . . • . . • . . . . . . . . . . • . . . . . . . . . . . . 118

Myce l i a l Growth . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1 8 S p orula t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121 Spore Ge r m i na t i on . . . . . . . . . . . . . . . . . . . . . . . . . . 1 2 5 TLC B i oa s s a y . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 27

D I S CUS S I ON AND CONCLUS I ON . . . . . . . . . . . . . . . . . . 1 3 1

iv

BIBLIOGRAPHY . . • . . • . . . . . . . . • . . . . . . . . . . . . . . . . 133 VI TA . . . . . . . .. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 3 7

v

L I ST OF TABLES

Tab l e

1 1H-NMR Data (6) o f (44) a nd GC1 . . . . . . . .

2 l3C-NMR Chem i ca l S h i f t Data (ppm )

P a ge

25

o f Gel................................. 32

3 13C-NMR Chem i ca l S h i f t Da t a (ppm ) o f F l i nders i ne w i t h the Observed Va l ues . . .

l 3C -NMR Chem i ca l S h i f t s (ppm ) o f

48

G l yc oma ur i n ( 2 2 ) ......... . ... . . . ....... 6 3

5 lH-NMR S pectra l Data (6) o f ( 2 2 ) , ( 5 1 ) a nd ( 5 2 ) . . . . . . . . . . . . . . . . . . . . . . . . 70

6 P l a n t Extra c t s I nves t i ga ted f or An t i f u nga l Act i v i t y . ...... . . . ... . .. . ... 1 1 3

7 E f fect o f G. cal c i co l a a nd G. r upes tr i s extra c t s o n Myce l i a l Growth of C. cucurbltarlum, C. capsicl a n d C . gl oeos por i o i des . . . .. . . . .... . . . .. . .. . 1 19

8 E'f fec t o f G. ca l c i co l a a nd G. rupes tr is ext r a c t s o n S pore Germ i na t i o n o f C . cucurbl tarlum, C . caps i c i a nd C . gl o e os pori o i des . . . .. . . . . .. . . . . . .... . 126

9 A n t i f unga l Cons t I t uent s o f E x t r a c t B • • • 129

v i

F i g u r e

1

2

3

4

L I ST OF FI GURES

Schema tic Dia g r am o f Extrac tion and I so l ation P r ocedure o f G . ca lc i cola (Lang k a wl I sland ) . . . . . . . . . . . . . . . . . . . . . .

I R S pe c t r um o f ( 4 4 ) ................... .

IH - NHR S p e c t r u m o f ( 4 4 ) ............... .

Mass S pe c t r um 0 f ( 4 4 ) . . . . . . . . . . . . . .. . . .

5 Mass S pe c t r a l Fragmenta tion P a t t e rns

P a ge

1 9

2 1

2 3

2 6

o f ( 4 4 ) . .. . . . . .... . . . . . . . . .. . . . . . . . . . . . 2 7

6 1 3 - 4 C NMR S pe ct r um o f ( 4} .............. .

7 PHSQC S pe c t r um o f ( 4 4 ) ................ .

8 PHSQC S p e c t r um o f ( 4 4 ) . . .. . . . .... . .... .

9 I R S pe c t r um o f ( 48 ) • • • • • • • • • • • • • • • • • • • •

10 Mass Spe c t r um of ( 48 ) ................. .

1 1 Mass S pe c t r al Fragmenta t i on Pa t t e rns

2 9

30

31

3 4

3 5

o f ( 48 ) ................................ 3 7

1 2 S chematic Dia g r am o f Ex t r ac t i on and I sola t i on P r oced ure o f G. calcicola (Temple r Par k ) . . . . . . . . . . . . . . . . . . . . . . . . . 3 8

1 3

1 4

15

1 6

1 7

1 8

1 9

Mass S pe c t r um o f ( 49 ) .. . . . . . . . . . . . . . . . .

I R S p e c t r um o f ( 49 ) ................... .

1 _ H NMR S p e c tr um o f ( 49) ............... .

NOESY S p e c t r um o f ( 49 ) ................ .

Cou p l ing o f Various P r o tons Obse rved f r om NOESY S p e ct r um o f ( 49 ) . . . • . . . • . . . .

1 3C -NMR S p e c t r um o f ( 49 ) .............. .

I R S pe c t r um o f ( 50 ) . . . . . . . . . . . . . . . . . . . .

v i i

40

4 1

4 3

4 4

45

4 6

4 9

20 IH-NHR Spe c t r u m o f (50) ............... .

2 1 Mas s S pe c t r um o f (50) ................. . 2 2 Mas s Spec t r a l Fragme n ta t i o n P a t t e r n s

51

5 3

o f (50)................................ 5 4

2 3

2 4

2 5

2 6

2 7

2 8

S chema t i c D i a gr a m of Extraction and I s ola t i o n P r ocedure o f G. r upes t r is S t em Ba r k ,a nd Leaves . . . . . . . . . . . . . . . . . . .

Ma s s Spec t r um of ( 2 2 ) . . . . . . . . . . . . . • . . . .

I R Spec t r um o f ( 2 2 ) ................... .

1�-NMR S p e c t r um o f ( 2 2 ) ............... . 1 3C-NMR S pe c tr um o f ( 2 2 ) . . . . . . . . .. . . . . .

Ma s s S p e c t r um o f (51) ................. .

2 9 Ma s s S p e c t r a l Fr a gme n ta t i on P a t t erns

5 6

58 5 9

6 0

6 2

6 5

o f (51) . . . . . . . . . . . . . . . ., . . . . . . . . . . . . . . . . 6 6

3 0 I R S pe c t r um of (51).................... 6 8

3 1 1H -NHR S pe c t r um o f (5 1). . . . . .. . . .. . . . .. 6 9

3 2 NOESY s pe c t r um o f ( 5 1 ) . . . . . . . . . . . . . . . . . 7 2

3 3 P r o t o n Cou p l i ng P a t t e r n Obs e r ved i n NOESY S pe c t r um o f (51)................. 7 3

3 4 Ma s s S p e c t r um o f (53).................. 7 5

3 5 I R S p e c t r um o f (53).................... 7 6

3 1 -6 H NMR S pe ct r um o f ( 5 3 ) . . . . . . . . . . . . . . . . 7 7

37 Mass Spectral Fragme n tat i o n Pat ter n s o f ( 53) . . . . . . . . . . . . . . ....... . ... . . ..... 7 8

3 8 I R S p e c t r um o f (5 4) . . . . .. . . . . . . . . . . . . . . 8 1

3 9 Mas s S p e c t r um o f (54 ).................. 8 2

4 0 H i s t o g r a m show i ng e f f e c t o f G. calclcola a nd G. r upest ris e xt r a c t s o n myc e l i al gr owth o f C. c u c u rb i tar i um, C. capslcl and C. gloeosporioides . . . . . . 1 2 0

4 1 H i s t o g r a m sho w i n g e f f e c t o f G . calcicola a nd G. r u�es tris e x t r a ct s on s po r ula t i on o f C . ca ps l c l and C. gloeosporlolde� . . . . . . . . . . ... . . . . . . . . 124

v i i i

LIST OF PLATES

P la te Page

1 Ch i l l i A i s a hea l thy f r u i t . Ch i l l i B sh owi n g s ympt oms o f a n thr a c n o s e . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 0 5

2 E f f e c t o f CHCl 3 e x t r a c t B o n myce l i a l g r owth o f Chaonephora cucurbi tar i um . . . . 1 2 2

3 Ef f e c t o f CHCl 3 e x t ract B o n myce l i a l g r owth o f Col l e tot r i chum caps i c i . . . . . . . 1 2 2

4 E f f e c t o f CHC 1 3 e x t rac t B o n myce l i a l g r owth o f Col l e totr i chum gl oeos por l o l des . . . . . . . . . . . . . . . • . . . . . . . . 1 2 3

5 E f f e c t o f CHCl 3 e x t rac t F o n s p o r e g e r m i na t i on o f Col l e t ot r i ch um caps i c i . . 128

6 CHC 1 3 e x t rac t B , GC7 (F l i nd e r s i ne (49» and Gce (de s me th oxyzanthophy l l i ne (50) w i th s ome i mpur i t i e s ) on TLC p late show i ng zones of i nh i b i t i on a f t e r s p r a y i ng w i th Asper g i l l u s fl a vus . . . . . . . 1 30

i x

br CHC13 CDC13 d d d d t EtOH FAB I R m m Me MeOH m.p. MS NMR PLC ppm s TLC w

LIST OF ABBREVIATIONS

br oad c h l o r o f or m d e ut e r a ted c h l or o f o r m d o ub l e t d o u b l e t o f d o u b l e t d o u b l e t o f t r i pl e t e t hano l Fa s t At om i c B ombar dment I n f r a Red med i um ( i n IR yata ) mult i ple t ( i n H -NMR d a ta ) me thyl me t hano l melting point Ma s s S p e c t r u m Nuc l e a r Magne t i c R e s onance P r e pa ra t i ve Th i n La ye r Chr oma t ogra phy pa r t per m i l l i on strong Thin Layer Chromatography weak

x

Abs t ra c t o f t h e t he s i s p r e s ented t o t h e Sena t e o f Un i v e r s l t i Pe r tani an Ma l a ys i a i n f u l f i l ment o f

r e q u i r e ments f o r t he d e g r e e o f Ma s ter o f S c i ence .

CHEMI CAL CONSTI TUENTS OF GLYCOSH I S CALCICOLA AND G. RUPESTRI S (RUTACEAE) AND THE I R B I OLOGI CAL

ACTI V I T I E S

Cha i r man : Fa c u l t y :

By

CHEW YEAN L I NG

June 1 9 9 5

A s s o c i a te P r o f e s s o r Dr . Ma va r d i Ra hmani S c i ence and Env i r o nment a l Stud i e s

I nves t i ga t i on o f the l e a f and t v i g s e x trac t s o f

Gl ycosm i s ca l c i col a ( R u taceae ) f r om t vo d i f f e r ent

l oca t i ons has l ed to t he i s o la t i on o f f i ve compound s .

The s t r u c t u r e s o f t h e s e compound s we r e e l uc i d ated by means o f s pec t r o s c o p i c methods ( I R , uv, I H-NMR , 1 3 C_

NMR , MS ) and by compa r i s on w i t h p r e v i o u s s t ud i e s.

G. ca l c i co l a c o l l e c t ed f r om Langk a v i I s l and

y i e l d ed �e t hy l ge r ambu l l i n I s i t o s t e r o l and anoth e r

unid�ntl f led c ompo und . From ano t her s e pa r a te

x i

collection of the same plant from Templer Park, a new

alkalo id (desmethoxyzanthophyll i ne) and f 1 i nders i ne

have also been isolated. A similar study on the bark

of another species, G. rupestris, collected from

Langkawi Island afforded

m e t h o xyg lyc o m a u r i n) ,

a new alkaloid (7-

g l yc o m a u r i n , d e s-N-

methylacronycine and stigmast-7-enol, while the leaves

of the plant gave an unidentified compound.

The antifungal activities of these plant extracts

were evaluated using the poison food and spore

germination techniques. The plant extracts exhibited

varying degree of fungitoxic response on the growth of

the fungal pathogens of chilli (Capsicum annuum L.).

The chloroform extract of G. calcicola collected from

Templer Park has been shown to be most effective in

inhibiting the mycelial growth, sporulation and spore

germination of Chaonephora cucurbi tarium,

Col1etotrichum capsici and C. gloeosporioides.

The antifungal compounds in this extract were

identified using TLC bioassay technique against

Aspergillus flavus. The results indicated that the

activity of the extract is due to the presence of its

major components: f lin d e r s i n e a n d

desmethoxyzanthophylline.

xii

Abs t rak t e s i s yang d ikemukaka n kepada S e na t Un i ve r s i t i P e r ta n i a n Ma l a ys i a ba g i memenuh i ke pe r l ua n

I j a za h Ha s te r Sa i ns .

KANDUNGAN KI HI A DARI GLYCOSHIS CALCICOLA DAN G. RUPESTRIS (RUTACBAE) DAN AKT I VI TI BI OLOGI NYA

O l e h

CHEW YEAN L I NG

J u n 1995

P e nge r u s i : P r o f e s s o r Had ya Dr . Ha wa r d i Rahma n i Fa k u l t i S a i ns dan Penga j i a n A l a m S e k i ta r

Kaj i a n k e a t a s e k s t r a k d a u n d a n r a n t i ng

Gl ycosmis ca l cico l a ( R utaceae ) t e l a h mengha s i lka n

l i ma k ompa un . S t r uktur s e ba t i a n t e r s e but te l a h dapa t

d i t e n t u k a n de ngan menggunaka n ka edah s pe k t r osko p i dan

pe r ba nd i ngannya d e ngan k a j i a n -kaj i a n ya ng t e l a h

d i l a k u k a n t e r d a hu l u .

G. calcjcola ya ng d l kump u l ka n dar i P u l a u

Langka wi te lah membe r l ka n me t i l g e r ambu l i n

s i t o s t e r o l d a n s a t u s e ba t l a n ya ng be l um d lke n a l

pa s t i . Dar i pengumpu l a n ber a s i ngan s pe s i e s ya ng sama

x i i i

di Templer Park, satu alkaloid baru

(desmetoksizantofilina) dan flindersina telah berjaya

dipencilkan . Kajian yang serupa ke atas kulit batang

spesis yang lain, G. rupestris, yang dikumpulkan dari

Pulau Langkawi telah member ikan satu alkaloid baru ( 7 -

metoksiglikomaurin), glikomaurin, des-N­

metilakronisin dan stigmast- 7-enol, manakala daunnya

pula member ikan satu sebatian yang belum dikenal

pasti .

Akti vi ti antikulat ekstrak tumbuhan tersebut

telah dikaji dengan menggunakan teknik 'poison food'

dan pereambahan spora. Ekstrak tumbuhan yang dikaji

menunjukkan aktiviti antikulat yang berbeza terhadap

pertumbuhan kulat patogen eili (Capsicum annuum L.).

Ekstrak klorofom G. calcicola yang dikumpulkan dari

Templer Park didapati paling berkesan mereneat

pertumbuhan misellum, pensporalan dan percambahan

spora Chaon ephora cucurbitarium, Colletotrichum

capsici dan C. gloeosporioides.

Sebatian antikulat dalam ekstrak ini telah dapat

dikenal pasti dengan menggunakan kaedah biocerakinan

TLC terhadap Aspergi 11 us f 1avus. Keputusannya

menunjukkan bahawa aktiviti ekstrak tersebut adalah

disebabkan oleh kehadiran komponen utamanya:

flindersina dan desmetoksizantofilina.

xiv

PART I

EXTRACT I ON AND I SOLAT I ON OF

CHEMI CAL COMPONENTS FR OM

GLYCOSH IS CALCICOLA

AND

G. RUPESTRIS

(RUTACEAE)

1

2

INTRODUCTION

Glycosm i s i s a genus o f f o r t y s pe c i e s o f sma l l

t r e e s and shr ubs o f the f am i l y Ru taceae , f ound mos t l y

i n S o uth-Ea s t As i a and S o uth o f Ch i na . F o u r t e e n

s pe c i e s a r e r e po r t e d t o occur i n Ma l ays i a ( Wh i tm or e ,

1 9 8 3 ) . G. calc i cola B.C . S t one and G. rupes t r i s

R i dl e y a r e shr ubs wh i ch g r ow o n l i me stone r oc k s and

l i me s t one h i l l s r e s pe c t i ve l y . G . calc i cola can be

f o und i n Lang k a w i , P e r a k , S e l angor and K e l antan

whe reas G . r upes tr i s occurs in P e r l i s , Kedah , P u l a u

Adang and Langkaw i .

Membe r s o f the genus a r e a r oma t i c and a r e used

med i c i na l l y , ch i e f l y the r o o t s ( B ur k i l l , 1 9 6 6 ). G .

penta phylla Co r r . i s used f o r the t r e a tment o f f ever

in India, vhile G. cochlnchlnensis Plr r e found i n the

Dutch I nd i e s

de c oc t i on o f

i s used t o t r e a t s w o l l en s p l een . A

the l e a ve s and r o o t s o f the l a t t e r ,

a l s o , i s used a s a s t i mu l ant t o d i ge s t i on .

3

L I TERATURE REV I EW

A numbe r o f stud i es o n t he c hem i c a l c o nst i t u e n t s

o f Gl ycos m i s spe c i es have b e e n r e por t ed . T h e spec i es

wh i ch have b e e n i nvest i ga t ed i n d e t a i l i nc l ude G.

pen taphy l l a, G . ma ur l t i ana, G . b l 1ocu l a r l s, G.

arborea, G. cyanocarpa a nd G. ch l or osperma . These

i nvest i ga t i o ns r e ve a l ed t h e presence o f va r i o us

c ompounds be l o ng i ng t o t he a l ka l o i d , c o umar i n a nd

a m i d e gro u ps .

One o f t h e p l a n t s t h a t had been e xhaust i ve l y

i nvest i ga te d was G . pen taphy l l a a nd i t f u r n i shes a

numbe r o f a c r i d on e , ca r ba z o l e a nd qu i n o l one bases .

The r o ot ba r k has be e n r e po r t ed t o e la b o r a t e t hese

a l k a l o i ds . I n 1 9 6 6 , Gov i ndacha r i e t . a l i so l a t e d a

known compound, sk i mm i a n i ne ( 1 ) , C1 4 H1 304 N , m.p .

1 7 60C a nd t hr e e new a c r i d on e a l k a l oids: n or a c r o nyc i ne

( 2 ) , C1 9 H1 7 03N , m . p . 200oC; des-N-me thyl a c r onyc i ne

( 3 ) , C1 9 H1 7 03 N, m . p . 2 68 - 2 7 0oC a nd des-N ­

me t hylnoracr o nyc i ne ( 4 ) , C1 SH1 5 03 N , m . p . 2 4 6 °C . I n

1969 Cha k ra bo r t y r e ported t he iso l a t i o n o f

g l yc o z o l i ne ( 5) , a

c a r ba zo le d e r i va t i ve .

Me.O

OMe. o OH

OMe

(1) (2)

o OR Me.O

(3) R = Me ( 5 ) R = H ( 4 ) R = H ( 6 ) R = OMe

I n con t i nuat ion o f their s t udies on

4

Me Me.

the

carba zol e a l k a lo i d s of G . pen taphyl l a, Cha kra bort y e t

a l . ( 1 9 74 ) re ported the s t ruc t ura l s t ud i e s o f

g l ycozo l i d i ne (6) , m.p . 1 6 0- 1 6 2 0C from the root bark

of th i s p l a n t . Subse que n t l y , Das and Chowdhury

( 1 9 78 ) d i scovere d a nove l

g l ycoso lone

q u i no l one a l k a l o i d ,

Further

5

s t udy o n the r o ot bark of th i s s pec i es by Chowdhury et

a l . ( 1 9 8 7 ) a f f o rded c a r ba z o le (8), C1 ZH1 9 N , m.p. 2 4 5 0C a nd 3 -methyca r ba z o le (9) , C1 3 H1 1N , m . p . 2 0 7 oC .

Th i s i s the f i rst repo r t o f t he iso la t i on o f c a r ba z o le

fr om a p l a nt source .

OMe.

( 7)

(8) R = H

( 9 ) R = Me

R

An a n t i ba cter i a l c a r ba z o l e a lka l o i d ,

g l yc o z o l id o l (10) , C1 4 H1 3 N02 ' m.p . 2 4 0oC was

d i sc o vered f r om the r o o t o f G. pen ta phylla by

Bha t tacha ryya et al . ( 1 9 8 5 ) . The c ompo und has been

f o und to be a c t i ve a ga i nst some Gra m-pos i t i ve a nd

6

Gram-negative bacteria. In the same year, he and

Chowdhury reported t he struct ure o f a n o t her carba z o l e

a lka o l i d , g l yc o z o l ida l ( II ) , m. p . l S50C . In 1 9 9 2 ,

Jash et a l . i so l a ted two car baz o l e

g l ycoz o l i c i ne (12) ,

source .

I H

(10) Rl = OMe; R2 = Me; R3 = OH

( II ) Rl = R 3 = OMei R 2 = CHO

(12) Rl = H; R 2 = Me; R 3

= OMe

(13) Rl = OMe; R2

= CHOi R 3 = H

a lkal o i ds ,

and

7

An i nve s t i gat i on o f the l eave s of G . pentapbylla

by Bhattachar yya a nd Chowd hur y ( 1 9 85 ) l e d t o the

i solat i on of a qu i n o l on e a l ka l o i d , g l ycol one ( 1 4 ), C 1 6 H 1 9NO) , m . p . IISoC . Fur t he r chem i ca l study on t he

l e ave s o f t h i s s pe c i e s by K a ma r u z zaman e t a l e ( 1 9 8 9 )

a f forded a c a r ba zole alk alo i d , mupami me (15),

C 1 9H 19N0 2 ' m .p. 15 2oC . Th i s i s t he f i r s t C I 8

a l k a l o id r e po r ted' from t h e genus Gl ycosm i s.

S a r k a r a nd Cha k ra bor t y ( 1 9 7 7 ) u nd e r t ook t h e

e xam i na t i 'on o f t h e f l owe r h e a d s o f G . pen taphyl l a

f r om wh i ch t h e y r e ported the i so l a t i on a nd s t r uc t u r e

o f g l ycophym i ne (16), C15 H1 2N 2 0 , m . p . 25 4 °C a nd a n

am i d e , g l ycom i d e ( 17), C1 4 H1 3N O , m . p . 1 2 0°C . I n 1 9 7 9 ,

they i s o l a ted a new qu i na zo l i ne a l k a lo i d ,

glycophym o l i ne ( 18 ) , C1 6 Hl 4N20 , m.p. l 650C .

Wor k on t he s e e d s o f G. pen taphyl l a by

Muk he r j e e et a l e ( 1 9 8 3 ) had y i e ld ed a car ba zo l e

d e rivative, glycozolinine ( 1 9) , m .p. 2 3l-2 3 2°C . From phys i ca l and che mic a l e v i d e nc e , i t s s t r uc t u r e wa s

i d e n t i f i e d a s 6 -h yd roxy - 3-me t hylc a r ba zole .

8

Me-O

o

Me.O Me.O

(14 ) (15)

o

(16)

(17)

9

o Me.. HO

�N -o� N�CH2 _

(16) (19 )

A chem i ca l i nv e s t i ga t i on o f t he r o o t s o f

G. maur i t i ana by Ras t og i e t a l . (1980) r e s u l ted 1n

t h e i s o l a t i on o f two a lka l o i d s :

I-hyd roxy-3-me t h oxy-2- ( 3-me t h y l bu t - 2-enyl ) -N-me thyl a c r i d a n -

9-one (20), m . p . 1 34 - 1 3 6o C a nd 4 , 8 -d i me t h oxy- 3- ( 3-

me t hy l but- 2 - e ny l ) -N -me thyl-2 -qu i n o l o n e ( 21 ) . The

s te m bark of G. ma u r i t i ana c o l l e c ted in G i r i ta l e ,

n o r t h c e nt r a l o f S r i La nka was s h own t o c o n t a i n

c a r ba zo l e a lka l o i d s : g l ycomaur i n ( 22 ) , m . p . 1 9 5 - 1 9 6oC

a nd g l ycoma ur r o l ( 23 ) , m . p . 1 4 9 - 1 50o C by Kumar e t a l .

(1989).

o OH

(20)

Me.O

( 21 )

10

I H

(22 )

(23)

A new 5 - hydroacr i d one wa s i s o l a t e d by Bowen e t

a l . ( 1 9 7 8 ) f r om t he l eaves o f G . bilocularis. It was

cha r a c te r i zed as 1 , 5 -d i hyd r oxy-2 , 3 -d i me t h oxy-10-

me thyl -9-a c r i d o ne (24 ), m . p . 2 0 6 - 20 7 oC . I n add i t i o n ,

t hey a l s o i s o l a t ed f ive k n own c ompounds: a r b o r i ne

(25) , m . p . 1 5 4 - 1 5 6 oC; a r bor i n i ne ( 2 6 ) , m . p . 1 7 4 -

1 7 6 0Ci s k i mm i a n i ne (1) , k ok us a g i nine ( 2 7 ) , m . p . 1 70 -

1 7 2 °C a nd g l yce r i ne (28), m . p . 1 4 5- 1 4 6 oC .