COUMARINS AND C I N S A J I I C ACID FROM G YMSOPH Y T O S ISA TIDICARP LiX REV&TORRES~, FRWCO DELLEMo\
XHE
and GIOI 4 \ \ 1 B L T T I S T MIRIN ~ BETTOLO
Ceirtro C S R , Ciramtca dea Recettorz e d e l l e ;IIolecole Bzologzcamente AtizLe, Istzizito dz Chzwizcn, 1 nztersztd Crittolzca d e l S a c r o Cztore, Ronia BRCCEK CASSELS Departantento d e
Qitinizcn,
Filcztltad d e Czeiiczn, rnazerszdnd Tdcnzca d e l Esiado,
Santiago, Chile
The Umbelliferae family is one of the best knoxm sources of couniarins (1). Lately, the antimicrobial activity of coumarins has been attiacting interest because of the posqible activity of these compounds as phytoalexins (2-5). I n the course of a systematic investigation of Chilean plants, we examined Gymnophytori isatidicarpunt, which g r o m in the vicinity of Santiago, Chile. I n screening for antimicrobial activity, a 50yc ethanol estract of that species shon-ed activity against Sarciiia lutea, Staphylococcus aureus and E . coli (6). From the petroleum ether extract, a considerable quantity of 6-methoxyi-geranyloxycoumarin (l), an unusual coumarin, \vas iqolated. This n-a. isolated for the first time by Larsen and Sandberg from the roots of T h a p s i a gargaiiica, another Umbelliferae species ( 7 ) and. then, from four other Rutaceous plants: Feroiiia elephanturn ( 8 ) , Haplophyllum kispaiiicunt (9), H . pedicellaturn ( l o ) , and Poncirus trifoliata (11, 12). ah-ay- in small quantities. From the methanol extract. t n o common coumarins, scopoletin (2) and esculetin (3) and their biogenetic precursor, cinnamic acid. were isolated. The bactericidal activity of Gymfzophyfoiz is due principally to esculetin. This coumarin has been b1~ou-nin a
previous report to be more bacteriostatic than simple phenols (5). I n our assays, 6-methoxy-7-geranyl-oxycoumarin n as not active against Staphylococcus pyogenes, E. coli, Pseudomopzas pyogenes, C a f i d i d a albicans, Streptococcus pyogeites and Diplococcus piieumoniae. The substance was tested in multistage assays which are convenient to demonstrate insect growth regulatory and insecticidal effects on three species: a bug ( D y s d e r c u s ciftgulatus), a mosquito ( C u l e x p i p i e n s )
R’oyJ-J*o \
R20
2
RI CHI CH,
3
H
1
Rz
Geranyl
H H
and a butterfly (Adomoplzyes orana). The results show a low insecticide activity on A . oraita and no insect growth regulatory effect.
EXPERIAIESTAL2 ~l.kTERI.IL.-G?7~t7IOp~l~fo12 isatid?carprcm (Presl. ex DC) Mathias Constance was collected in December (summer) 197G in San JosC dei Maipo near Santiago (prov. Santiago, Region Metropolitans, Chile). A
PL4ST
W e l t i n g points n e r e obtained in a Xofler apparatus and a r e uncorrected: uv spectra were recorded on a Beckman model Acta I11 spectrophotometer; p m r spectra were determined on a T‘arian model E l 1 360 instrument, using TMS as internal s t a n d a r d ; mass spectra were run on an LKB-9000 S mass spectrometer.
_____ 'Fellowship from I I L i C S R . On leave
from Facultad de Ciencias Cniversidad TBcnica del Estado, Santiago, Chile.
532
SEP-OCT
19T9]
TORRES ET AL. :
COCIIARISS ISD
CISS-UIIC
voucher specimen is kept in t h e Herbarium of t h e 1Iuseo Sacional de Ilistoria S a t i i r a l , Santiago, Chile.
ACID
3:3
solid n-hich cr>-stallized from chloroform. I t had a nip 204-205'. The p h y i c a l and spectral d a t a n-ere identical lvith a n 311thentic sample. EXTK\CTIOS .\ID ri:.\~~~os.\~~os.~Finely g r < ~ r i ~aerial id p a r t s i5.5 kgi of G . isotidiCIzs.nfIc .\CID .-Flirt her elution \Tit h c*irpitiii were extracted with petroleum e t h e r cliloroiorni gave a n-Lite solid crystallized and then with methanol in R Soxhlet appafrom chloroform, nip 133'. Physical and ratus. The petrolemi e t h e r extract 110 spectral d a t a of t h e acid and i t s nieth>-l l i t e r s ) \\-as concentrated t o 2.5 liters and let ester \\-ere identical with those of authentic samples. s t a n d at 0" for 48 h . -1ivhitish yellon- solid was obtained (solid 67.5 g ) . T h e niethaESCL-LETII 131 .-Elution with chloroform nolic extract (10 liters) 1%-asconcentrated t o yielded 60 mg of a solid m-liich m-as crystal1 liter and 4 liters of water were added. lized from chloroform, nip 265-66". Spectral T h e mixture JTas let stand a t 0' for 24 h. d a t a x e r e identical lvith those of a n aiithenThe solntion was filtered and extracted tic sample. with chloroform. When t h e chloroform la>-ers \\-ere washed with xwter, dried n-ith -1CKSORLED1IEST anh>-drous Sn2SOs, filtered, and concenR e are indebted t o Dr. -1.G. N a a g , t r a t e d t o (dryness, a brownish residue fresiDielsdorf, Switzerland, for t h e biological due B , 30 g) resulted. tests on insects. CH.\RICTERIZ.LTIOS O F 6-JIETHOST-7-GEIlRecei;,ed 27 dIarcli 1979. . L S T L ~ I T C O C ~ I . ~ R I S (1) .-Tlc showed just one fluorescent spot for residue A (1 g ) LITER.1TURE C I T E D which was purified b y means of column 1. For a reviem see e.g. a ) -1.G Gonzalez, Chromatography 120 g, silica gel) prepared R . J Cardona, H . Lopez, J. 11. l l e d i n a in benzene. Elution with chloroform afand F. Rodiguez Luis, Re;. Real .lead forded 750 nig of a white solid which crystalCienc , 70, 109 11976) h ) R . Hegnauer, lized from ethanol. I t gave t h e follon-ing Ombelliferes Copitrib. Plztridiscip. Sgst physical properties: mp 88-89" (lit. 18) nip -4ctes Siwib. I n t . 2nd. 335-63 11977) 88-89"): iiv, Xmas (log e) ( E t O H ) , 229, 295 2. C. Johnso; and D. R Brannon, Phgtoand 345 n m i4.17. 4.56 and 4 . 5 0 1 : i r . miax chenristr), 12, 2961 (1973). I K B r ) , 1715, lGl0: 1560 em-': p m r (CDCI,) 3. J . L. King, -1.D. Alihara, K. Corse and 6 1.61 id, 3H, J = l H z ) , 1.65 (d, 3 H , J = l H . Bayne. Phgfocheniistrg, 10, 2965 H z ) , 1.76 (d, 3H, J = l H z ) , 2.0-2.25 im. (1971). 4H), 3.83 is. 3 H ) . 4.8X1.32 im. 1 H ) . 4.71 4 . Y. D a d a k and K. Hodak. E.tberienfia. ( d J ' 2 H , J=6'.5 H z j , 5.47 (m, lH,' J=G'.5 Hz -- ~ - - and J = l H z ) , 6.26 (d, l H , J = 9 H z ) , 6.82 5 . F. C . Fisher and A. B . Svendsen, Pligfo(s, l H ) , 6.85 (s, l H ) , 7.63 (d, l H , J = 9 . 5 H z j ; chewistry, 15, 1079 (1976). ms, ni,'e 328.1667 ()I-, calculated for 6. D . S.Bhakuni, 11. Bittner, C . MartiCzaH,aO,, 328.1671 (111, 259 (101, 193 ( 2 2 ) , corena. AI. Silva. E. R e l d t , M.E . l I e l o 192 (1001, 191 il5), 177 (12), 164 (G), 119 ( 5 ) , and R'. Zemelman, L l o y d i a . 37. 621 136 ( l G ) , 93 110). il9741. These spectral d a t a were identical with I . P. K. Larsen and F. Sandberg, d c t u those already reported (8). Acid h>-drolysis Ciiem. Scnnd., 21, 113 (1970). in 2s-methanolic HC1 produced scopoletin, 8. S.K. Talapatra, 11.IC. Chaudhuri and nip 204-205", identical in all respects with an B . Talapatra, Phgiociieniistry, 12, 236 authentic sample. (19T3). 9. .1. G. Gonzales. R . J . Cardona. R . SEP.LR.LTIOS OF COJIPOCSDR O F 3IETH.iSOLIC Xorena and F. Rodriguez Luis, - 4 t i . ExTR.Lc'r.--Residue B (3 g ) , x-hich showed Q U I I U69, . 781 (19731. three principal spots in tlc, was separated b>10. G . -1.Hiiznetsova and S . F. Gashimov, column chromatography. T h e column was Khzvi.Prir. Soedzn, 666 (1972). prepared n-ith 300 g of silica gel with benzene 11. -1. Guiotto. P. hodiehiero and U. and was eluted m-ith benzene, benzeneFornasiero, ' Z . i \ h f i ~ ~ f ~ r s c i i .29c, , 201 chloroform mixture, chloroform and chloro11974). form-methanol mixtures. 12. A . Guiotto, P. Rodighiero, G. Pastorini SCOPOLETIS (2).-Elution with benzeneand E . Celon, Piigfociieniistrg, 16, 1257 chloroforni c1:lj yielded 550 mg of a white (1977). ~~
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1
