Departamento de Quimica, Institute Tecnol6gico y de Estudios Superiores de Monterrey, Monterrey, N . L., Mixico
MEDICINAL PLANTS FROM MEXICO PART XXII ISOLATION O F ZALUZANIN-C A CITOTOXIC SESQUITERPENLACTONE F R O M ZALUZANIA PARTHENOIDES
Abstract From a petrol extract o f Zaluzania parthenoides, the citotoxic guaianolide zaluzanin-C was obtained. Besides B-sitosterol and stigmasterol were separated on column chromatography.
Introduction
In the southern mexican folk medicine, the Zalzrzania augusta known as ,,hierba blanca" and related species have been used for treating stomach ailments (MART~NE 1959). Z From Z . augusta, three sesquiterpenlactones, zaluzanina A, B and C were isolated (ROMOet al. 1967), from another species, 2. triloba, two sesquiterpenlactones, zaluzanin C and D were obtained (ROMOVIVARet al., 1967). Recently COLEet a]., 1974) in the screening for plants having tumor-inhibiting constituents, found that ethanolic extract of the whole plant Zaluzania robinsonii has inhibitory activity toward P-388 lymphocytic leukemia and were able to trace this citotoxic activity t o zaluzanin-C. In the present study, the 2. parthenoides was selected because of the popular use of it as a tea in northern Mexico for stomach activity. From the petrol ether extract of the aireal part of the plant a precipitate was obtained from which on recrystallization from acetone there were obtained white plates of zaluzanin-C CISH180s (M' 246) mp. 93-95', (a) 37", the infrared spectrum exhibited the typical y-unsaturated lactone absorptions at 1974 and 1239.' and two bands for C = C unsaturation a t 1639 and 900 cm-'. T h e NMR spectrum showed the characteristic (YOSHIOKA et al., 1974) two low field doublets a t 6.21 and 5.52 (2H. J=3Hz) corresponding to the hydrogens of an exocyclic methylene conjugated with the y-lactone, two singlets a t 5.03 and 4.97 (2H) corresponding to an exocyclic methylene group attached to C-10. Besides two triples suggesting an allylic
+
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By X. A. DOM~NGUEZ, J. M A R R O Q U AND ~ N E. CARDENAS
Dominguez, Marroquin and Cirdenas
Planta medica Vol. 28 1975
coupling a t 5.45 and 5.35 (2H, J= 1Hz) corresponding t o an exocyclic methylene group attached a t C-4. Also a triplet centered a t 4.15 (lH, 9 =9Hz) belongs t o the hydrogen joined t o C-4 which bears the ethereal oxygen of the lactone. In the region 2.95-1.1 there were several multiplets corresponding t o 10 hydrogens from methylene and methyne groups. T h e absence of methyl groups singlets eliminated the posibilitL of dealing with a pseudogeiaianolide lactone. O n checking the literature the search pointed to zaluzanin-C as the identity of.our compound. This was verified by comparison with an authentic sample and formation of its acetate. Besides the medical significance of this compo.und it seems that zaluzanin-C also is a chemical taxonomic marker. A comparativa T L C of petrol ether and ethanolic extracts failed t o show the other known zaluzanins. Not alkaloids were detected 1973). O n silicausing modified Dragendorff as chromogenic agent (DOM~NGUEZ gel chromatography of the petroleum ether extract, octacosane, 0-sitosterol and stigmasterol were isolated.
Experimental The IR spectra (KBr discs) were taken on a Perkin Elmer model 137, the optical rotation on a polarimeter Perkin Elmer model 141. The NMR spectra were recorded with a Varian A-60A in CDCI,; shifts are expressed as S values (parts per million) from TMS as internal standard. M. Ps. are incorrected and taken on a Kofler hot stage. Microanalysis were performed by the Alfred Bernhardt laboratories from Engelskirchen, Germany. The TLC were run on Merck silica-gel G.
Isolation procedure. One kilogram of dried and grounded aereal part of Z. parthenoides were Soxhlet extracted exhaustively with petroleum ether (b.p. 30-60"). On concentration of the extract to half volume, a crop of crystalline material (0.95 g) was collected, the filtrate was evaporated yielding 32.5 g of a greenish resin. The crystalline material was twice recrystallized from acetone-hexane (9 :1) affording 650 mg of white plates mp. 93-95' (Romo et al. 1967) gave 95-96'. (Found: C, 73.02, H, 7.30: 0, 19.35; Calcd. for C,,H,,O,; C, 73.15; H, 7.37; 0, 19.48%). 36.2"; (a) -I- 38.2; (a) 548 45.2; (a) -t- 68.6; ( u ) 90.0; (a) 63.8'; conc. (a) 20 mg in CHCI,.
,,,+
,,,
+
,,,
,,,+
,,, +
IR, T 3525 (OH).
NMR, N o depression was observed on mixed mp with an authentic specimen. Co-TLC with benzeneethyl acetate (8 :2) one spot.
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
90
Isolation of Zaluzanin-C
91
Zaluzanin-C acetate, A mixture of 115 rng of zaluzanin-C, 2 rnl acetic anhydride and 1ml pyridine was refluxed 2 hs. O n water dilutation and recrystallyzation 105 mg of white crystals were obtained. mp 102-104°. IR 1758 (y-lactone) and 1728 cm-I (acetate). NMR, singlet at 2.04 (3H). The petrol ether extract was chromatographed on silica-gel column. The elution was run with varios solvents, starting with benzene, with increasing order of polarity. From benzene-chloroform fraction l.60 mg of octaeicosane, mp. 64" C were obtained. Mixed mp. IR and NMR evidence. T h e chloroform fractions afforded 100 mg of crystalline p-sitosterol, mp. 138-140°, (a - 3S0, mixed mp., NMR. Acetate rnp., 136-128'. From latter fractions 63 mg of stigmasterol, mp. 165-168", (a) - SO0; mixed mp., IR, NMR acetate mp. 140.
,,,
References DOM~NGUEZ, X. A.: ,Mttodos de Investigaci6n Fitoquimica" Lirnusa, Mtxico (1973) J. R.: J. Pharm. Sci. 60,1321 (1974) JOLAD,S. D., WIEDHOF,R. M. and COLE, MART~NEZ, M.: ,Las Plantas Medicinales de MCxico", 4a ed. Editorial Botas, Mkxico p. 430 (1959) P.: Tetrahedron 23,29 (1967) ROMO,J., ROMODE VIVAR,A. and JOSEPH-NATHAN, ROMODE VIVAR,A., CABRERA, A., ORTECA,A. and ROMO,J.: Tetrahedron 23, 3903 (1967) H., MABRY,T. J. and TIMMERMANN, B: N.: ,Sesquiterpene lactonesu, Universiy of YOSHIOKA, Tokio, Press Tokio (1974) Address: Dr. Xorge A. Dominguez, lnsiituto Technologico y d e Estudos Superiores d e Monterrey, Chemistry Department, Monterrey, N. L., Mexico
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
,,,
Acknowledgements. T o Dr. JES~JS ROMO, Institute Quimica U. N. A. M. for a sample of zaluzanin-C. T o Dr. TOM MABRY,University of Texas for mass spectra. To. Lic. G. BUENO ZURIONand Dr. ONDARZA for their interest on this research. T o CONACYT for financial support, research grant 015.
MEDICINAL PLANTS FROM MEXICO PART XXII ISOLATION O F ZALUZANIN-C A CITOTOXIC SESQUITERPENLACTONE F R O M ZALUZANIA PARTHENOIDES
Abstract From a petrol extract o f Zaluzania parthenoides, the citotoxic guaianolide zaluzanin-C was obtained. Besides B-sitosterol and stigmasterol were separated on column chromatography.
Introduction
In the southern mexican folk medicine, the Zalzrzania augusta known as ,,hierba blanca" and related species have been used for treating stomach ailments (MART~NE 1959). Z From Z . augusta, three sesquiterpenlactones, zaluzanina A, B and C were isolated (ROMOet al. 1967), from another species, 2. triloba, two sesquiterpenlactones, zaluzanin C and D were obtained (ROMOVIVARet al., 1967). Recently COLEet a]., 1974) in the screening for plants having tumor-inhibiting constituents, found that ethanolic extract of the whole plant Zaluzania robinsonii has inhibitory activity toward P-388 lymphocytic leukemia and were able to trace this citotoxic activity t o zaluzanin-C. In the present study, the 2. parthenoides was selected because of the popular use of it as a tea in northern Mexico for stomach activity. From the petrol ether extract of the aireal part of the plant a precipitate was obtained from which on recrystallization from acetone there were obtained white plates of zaluzanin-C CISH180s (M' 246) mp. 93-95', (a) 37", the infrared spectrum exhibited the typical y-unsaturated lactone absorptions at 1974 and 1239.' and two bands for C = C unsaturation a t 1639 and 900 cm-'. T h e NMR spectrum showed the characteristic (YOSHIOKA et al., 1974) two low field doublets a t 6.21 and 5.52 (2H. J=3Hz) corresponding to the hydrogens of an exocyclic methylene conjugated with the y-lactone, two singlets a t 5.03 and 4.97 (2H) corresponding to an exocyclic methylene group attached to C-10. Besides two triples suggesting an allylic
+
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
By X. A. DOM~NGUEZ, J. M A R R O Q U AND ~ N E. CARDENAS
Dominguez, Marroquin and Cirdenas
Planta medica Vol. 28 1975
coupling a t 5.45 and 5.35 (2H, J= 1Hz) corresponding t o an exocyclic methylene group attached a t C-4. Also a triplet centered a t 4.15 (lH, 9 =9Hz) belongs t o the hydrogen joined t o C-4 which bears the ethereal oxygen of the lactone. In the region 2.95-1.1 there were several multiplets corresponding t o 10 hydrogens from methylene and methyne groups. T h e absence of methyl groups singlets eliminated the posibilitL of dealing with a pseudogeiaianolide lactone. O n checking the literature the search pointed to zaluzanin-C as the identity of.our compound. This was verified by comparison with an authentic sample and formation of its acetate. Besides the medical significance of this compo.und it seems that zaluzanin-C also is a chemical taxonomic marker. A comparativa T L C of petrol ether and ethanolic extracts failed t o show the other known zaluzanins. Not alkaloids were detected 1973). O n silicausing modified Dragendorff as chromogenic agent (DOM~NGUEZ gel chromatography of the petroleum ether extract, octacosane, 0-sitosterol and stigmasterol were isolated.
Experimental The IR spectra (KBr discs) were taken on a Perkin Elmer model 137, the optical rotation on a polarimeter Perkin Elmer model 141. The NMR spectra were recorded with a Varian A-60A in CDCI,; shifts are expressed as S values (parts per million) from TMS as internal standard. M. Ps. are incorrected and taken on a Kofler hot stage. Microanalysis were performed by the Alfred Bernhardt laboratories from Engelskirchen, Germany. The TLC were run on Merck silica-gel G.
Isolation procedure. One kilogram of dried and grounded aereal part of Z. parthenoides were Soxhlet extracted exhaustively with petroleum ether (b.p. 30-60"). On concentration of the extract to half volume, a crop of crystalline material (0.95 g) was collected, the filtrate was evaporated yielding 32.5 g of a greenish resin. The crystalline material was twice recrystallized from acetone-hexane (9 :1) affording 650 mg of white plates mp. 93-95' (Romo et al. 1967) gave 95-96'. (Found: C, 73.02, H, 7.30: 0, 19.35; Calcd. for C,,H,,O,; C, 73.15; H, 7.37; 0, 19.48%). 36.2"; (a) -I- 38.2; (a) 548 45.2; (a) -t- 68.6; ( u ) 90.0; (a) 63.8'; conc. (a) 20 mg in CHCI,.
,,,+
,,,
+
,,,
,,,+
,,, +
IR, T 3525 (OH).
NMR, N o depression was observed on mixed mp with an authentic specimen. Co-TLC with benzeneethyl acetate (8 :2) one spot.
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
90
Isolation of Zaluzanin-C
91
Zaluzanin-C acetate, A mixture of 115 rng of zaluzanin-C, 2 rnl acetic anhydride and 1ml pyridine was refluxed 2 hs. O n water dilutation and recrystallyzation 105 mg of white crystals were obtained. mp 102-104°. IR 1758 (y-lactone) and 1728 cm-I (acetate). NMR, singlet at 2.04 (3H). The petrol ether extract was chromatographed on silica-gel column. The elution was run with varios solvents, starting with benzene, with increasing order of polarity. From benzene-chloroform fraction l.60 mg of octaeicosane, mp. 64" C were obtained. Mixed mp. IR and NMR evidence. T h e chloroform fractions afforded 100 mg of crystalline p-sitosterol, mp. 138-140°, (a - 3S0, mixed mp., NMR. Acetate rnp., 136-128'. From latter fractions 63 mg of stigmasterol, mp. 165-168", (a) - SO0; mixed mp., IR, NMR acetate mp. 140.
,,,
References DOM~NGUEZ, X. A.: ,Mttodos de Investigaci6n Fitoquimica" Lirnusa, Mtxico (1973) J. R.: J. Pharm. Sci. 60,1321 (1974) JOLAD,S. D., WIEDHOF,R. M. and COLE, MART~NEZ, M.: ,Las Plantas Medicinales de MCxico", 4a ed. Editorial Botas, Mkxico p. 430 (1959) P.: Tetrahedron 23,29 (1967) ROMO,J., ROMODE VIVAR,A. and JOSEPH-NATHAN, ROMODE VIVAR,A., CABRERA, A., ORTECA,A. and ROMO,J.: Tetrahedron 23, 3903 (1967) H., MABRY,T. J. and TIMMERMANN, B: N.: ,Sesquiterpene lactonesu, Universiy of YOSHIOKA, Tokio, Press Tokio (1974) Address: Dr. Xorge A. Dominguez, lnsiituto Technologico y d e Estudos Superiores d e Monterrey, Chemistry Department, Monterrey, N. L., Mexico
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
,,,
Acknowledgements. T o Dr. JES~JS ROMO, Institute Quimica U. N. A. M. for a sample of zaluzanin-C. T o Dr. TOM MABRY,University of Texas for mass spectra. To. Lic. G. BUENO ZURIONand Dr. ONDARZA for their interest on this research. T o CONACYT for financial support, research grant 015.
