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Triterpenes from Simaba multiflora
P/ym&miwy. Vol. 26, No. 12, pi. 3301-3303. 1987. Printed in Great Britain.
TRITERPENES
0
003 I -9422/87 f3.00 + 0.00 1987 Pmgamon Journala Ltd.
FROM SZMABA MULTZFLORA
MUNEHISA ARISAWA, AKIO FUJITA, NAOKATA MORITA, PHILIP J. Cox; CORDELL~ Facultyof Phamuuxutical Sciences, Toyama Medical and P -tical of Pharmacy, Robert Gordon’s Institute of Technology, Schoolhill,
R. ALAN How&t
and GEOFFREY
A.
University,2630 SugitaniToyema 930-01, Japan;*School
Aberdeen, AR9 IFR, Scotland; tChemistry Department, University of Aberdeen, M&on Walk, Old Aberdeen, AB9 2UE. Scotland; $Program for Collaborative Restarch in the Pharmaceutical Sciences, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois 60612, U.S.A. (Revised receiwd 6 April 1987)
Key Word Imdcx-Simaba mulfifloro; Simaroubaaae; canthin-6-onc; 8-0-methylretusin; vanillic acid; hispidol B; 3$23R,25-trihydroxytirucall-7un-24-one; X-ray analysis.
Abstract-A new triterpene has been isolated from Sin&a multijba (Simaroubaceae) and determined to have the structure 3S23R.25-trihydroxytirucall-7-en-24-one (1). Hispidol B was confirmed to be 3S,23S,24R,25tetrahydroxytirucall-7-ene (2) by X-ray crystallographic analysis. Canthindane, 8-O-methyl retusin and vanillic acid were also obtained in the course of this work.
lN’fRODUCTlON
Sima6a multifira A. Juss. (Simaroubaceae) has previously afforded several quassinoids [l-4], canthin&ones [4,5], canthin-2,6diones [S], coumarins [4,5], a coumarinolignan [6], two phenolic compounds [4] and j?-sitosterol+~ glucoside [2]. The quassinoids are of interest because of their anticancer activity, and biogenetically the skeleton is derived from a tin&l-7ene derivative by way of an apoeuphol rearrangement [7,8]. However, tirucall-7cne derivatives have been obtained only occasionally from Simaroubaceous plants [9-121 and we report here on the isolation and structure determination of a new member of this series.
these two protons in 4 and the negative CD curve for 1, the stereochemistry of C-23 [15] was assigned as R. Consequently, the structure of 1 was assigned as 3&23R,25-trihydroxytirucall-7-eu-24-one. Compound 2, mp 252-254”, was found to be the known compound hispidol B, whose structure was previously assigned in ref. [14]. In order to thoroughly establish the stereochemistry in the side chain. X-Ray crystallographic analysis of this compound was performed, through which its structure was wniirmed lo be 3S,23&24R,25tetrahydroxytirucall-7-e. Canthindane [ 161 and 8-Omethylretusin [17] were identified by comparison with spectral data. Vanillic acid was identified by direct wmparison with an authentic sample.
RESULTS AND DISCUSSION
In a continuing search for anticancer agents from the wood of Simaba multipora [2, 4-61, a new triterpene was isolated and determined to be (3&23R,25)-hihydroxytirucall-7en-24-one (1). Hispidol B (2) was also obtained and confirmed lo be 3S,23&24R,25tetrahydroxytirucall-7-ene by X-ray analysis. Compound 1, mp 208-213”. molecular formula C,,H,,O, by high resolution mass spectrometry. exhibited IR absorption bands at 3500, 3250 (OH) and 169Ocm-’ (GO). The ‘H NMR spectrum of 1 indicated the presence of seven tertiary methyl groups, one secondary methyl group, two multiplets for oxymethine protons and one o1efGc proton. These signals were regarded as being characteristic of the tirucall-7cne system having a 3-hydroxy group [12-141. A singlet for two methyl groups shifted downfield to 6 1.30 (6H) suggesting attachment to an oxygenated carbon, i.e. C-25. An oxymethine proton was shifted to 63.83 indicating the presence of a vicinal diol, and an ion at m/z 459 [M - Me] + for loss of the C14 methyl supports the assignment of a double bond to C7. LAH reduction of 1 afforded 3, the C-23, C-24 isomer of 2. in which both the 23-H and 24-H were shifted substantially upfield. From the coupling constant between 3301
R’
b
R’
R’
0
H
OH
H
/OH ‘I/,~ <”
“OH /OAc
H
OH H
0:
H
““H
OAc
H
AC
/OAc ” “H
H
OAc
AC
M. ARISAWAet al.
3302 EXPERIMENTAL
(CDCI,): 60.75 (3H, s, 19-H,), 0.83 (3H, s, 18-H,), 0.86 (3H, s, 3@ H,), 0.97 (6H, s, 28 and 29-H& 1.03 (3H, d, J = 5.8 Hz, 21-H,), Mps were determined by means of a hot plate and are uncorr. 1.30 (6H. z, 26 and 27-H,), 3.25 (1H. dd, J = 5.3,10.9 Hz, 3a-H), ‘HNMR~od”CNMRspcctrowercr~rdedat200MHzmd 3.28 (Hi, d, J = 5.7 Hz, 24-H), 3.83 (lH, ddd, J = 3.2,5.7,5.7 Hz, 50.3 MHz, respectively (TM& int. standard, chemical shifts: 6). 23-H) and 5.26 (lH,dd. J = 0.8.2.2 & 7-Hk MS m/z: 476 [hi) l, Prelimialuy p?oces.wng end isolation. Details coaceraing the 461, 443, 425, 406, 4OQ 386, 371, 353. 327 and 309; Mass collection, identification and prelimimuy fractionation of the measurement,obs.: 476.3858. c&. for Cz0HSz04, 476.3863. wood of Sin&a muIripor0 A. Juss. (Simaroubaaac) have been Aceryfation of3. The reduction product 3 (1 mg) was dissolved in described previously [2,4-6]. In a oontinution of the previous A@-pyridine (1: 1, 0.3 ml) at room temp. overnight, dried In study [6], the mother liquor after the isolation of emodin was wrcuo and chromatographed over silica gel to alford a triacemtc 5, rechromatogtapbed on silica gel eluting with hexane-EtOAc (15) mp 152-155” (MeGH). ‘H NMR (CDC&): 60.76 (3H, s, 19-HJ). to afford I-0-methylretusin (10 mg), and anthiod-one (3 mg). 0.79 (3H,s, la-H,),0.85 (3H,s, 30-H&0.93 (3H, s, 29-H,),0.94 Droplet countercurrent chromatography (DCCC) of the com(3H,d,J = 5.1 Hx,21-H,),O.% (3H,s.28-H,), 1.20(3H,s,26or bined fractions 11 and 12, using a biphasic mixture of 27-H,), 1.25(3H,s,27or2~H,A4.51 (lH,dd,J = 5.3.9.3 Hx,3aCHClx-CeHe-MeGH-Hz0 (3: 1: 5:2) afforded t&in, 8- H), 4.93 (1H. d, J = 2.2 Hx 7-H), 5.23 (1H. m, .I = 3.6 Hx, 23-H) hydroxycanthi&-one and simalikalactone D as previously and 5.24 (lH, m, 24Hk MS m/z: 602 [Ml’, 587,569, 542,527. reported [6], and hispidol B (2 12mg), 3S,23R,25- 509,484,482.467,449.425,4C9,407.389.369,353,325 and 309. trihydroxytirucall-?c-24-one (I, 5 mg) and vanillic acid (6 mg). Crysrul data of 2. C,,H,zO., M = 476.4. ~horhombi~ u 3S~3R,25-Trih~~oxytdroxytinccaif-7-en-24-one (1). Recrystaliixed = 10.605(39), b = 14.455(75), c = 18.203(31) A, U = 2790.4A3, from Me&O gave colourless needles, mp 208-213”. CD: [6 Jz9, Z = 4, 0, = 1.13 8ctK3, F(OO0)= 1056, MO-K. radiation, i - 3330” (CHCl,k IR vzcm -I: 3500, 3250. 2950, 2910, 1690, = 0.7108 A p = OAOcm-‘. Space group F&2,2,. 1460,1450,1380,1360and 1060; ‘H NMR (CD&): 60.83 (3H. s, Crystalfagrophic analysis of 2. The X-ray intensities were 18-H,), 1.01 (6H. s, 19-Ha and 30-H& 1.03 (3H. d. J = 6.5 Hr, collated on a Nicolet P3 automated diffractometer using mono21-H,), 1.05 (3H, s, 29-H,), 1.12 (3H. s, 28-H,), 1.30 (6H, s, 26-H, chromatixul MoK, radiation. Integrated relative intensities for and27-Hxk2.69 (lH,m,22-Hk3.28 (lH,m,3-Hk3.83 (lH,m,231375 independent reflexions with 20 < 40” were measured as BH) and 5.32 (1H. m. 7-H); MS m/z: 474 [Ml’. 459 [M - Me]‘, 20scans; all reflexions were used in subsequent calculations. The 441 [M - Me-HzO]+, 423 [M - Me- 2HzO]‘, 369, 351, 325. structure was elucidated using MITHRIL [la] and the Hatoms 271,189,121,72and 55. Mass measurement, obs.: 474.3697. a&. were observed in electron density mapscalculated at intermediate for Cx,H,eO,, 474.3706. stages of structure refinement. The co-ordinates for allatoms and Hizpidol B (2). Recrystallimd from MeGH gave colourless anisotropic thermal parameters for the non-hydrogen atoms needles, mp 252-254”. IR$&cm-i: 3340, 2950. 1450, 1370, were varied in least-squares calculations using SHELX [ 191.The 1160,1070,1030,990,825 and 795; ‘H NMR (pyridine-d,): 60.83 C-H and GH bonds were constrained to be qua1 to 1.00 A and (3H,~l8-H~~O.~(3H,~l9-H~~l~O2{3H,s~~H~~l.l2(3H,~ the hydroxy, methyl and remaining hydrogen were given 29-H,), 1.14 (3H.4 .J = 6.8 Iix, 21-H,), 1.18 (3H, s, 28-H,), 1.62 common temperature factors during refinement. Unit weights (3H, s, 26-H,), 1.64 (3H,s, 27-H,), 1.88 (2H,m, 6-H,), 2.35 (lH, were employed and refinement converged at R 7.2%. Final dd, J = 9.5, 12 Hz, 22-H), 3.49 (lH, dd, J = 8.0 Hx, 31x-H),3.68 positional parameters are listed in Table 1, bond lengths in (lH,br~2~HA4.~(lH,~~ = 4.~9.5~2~H~S.~(lH,~7Table 2, valency angles in Table 3 and torsion angles in Table 4. H),5.53,5.74,5.89and6.08 (each, lH,m,excbangeablewith D,O, Thermal parameters are listed in a Supplementary Publication. OHkMSm/z:476[M]+,461[M-Me]+.458[M-H,0]*,433 [M -Me - HzO]+, 425 [M - CH, -2HzO]+. 407, 400. 385. Acknowledgements--This work was supported, in part, by 372, 353 and 327. Mass measurement, ohs.: 476.3838. talc. for grant CA 20164 from the Division of Cancer Treatment, CxOH>zO*, 476.3866, “CNMR (pyridine-d,): 13.38 (q, 18-Q National Cancer Institute. The authors would also like to thank 15.53 (q, 29-Q 18.37 (t, 11-C), 19.46 (q, 19-C), 24.35 (t, 6-Q 27.04 Professor J. R. Cole at the University of Arizona. for a gift of an (q, 26-Q 27.43 (q, 3&C), 27.66 (q, 27-C), 28.24 (q, 28-C), 2836 (t. authentic sample of hispidol B (2) and Mr M. Morikoshi. 16-Q 28.72 (t, 2-Q 34.13 (t, 12-Q 34.26 (t. 15-Q 34.26 (d, 20-Q Toyama Medical and Pharmaceutical University, for measure35.12 (s, l&C), 37.57 (t, l-C), 39.44 (s, 4-C), 42.03 (t, 22-C). 43.77 (s, ment of the 13CNMR and mass spectra. 13-Q 4925 (4 9-C) 51.07 (d, 25-Q 51.43 (s, M-CA 54.28 (4 17cX 69.29 (d, 23-C), 73.69 (s, 5-C), 76.50 (d, 24-Q 78.20 (d, 3-Q 118.32 (d, 7-C)and 145.95 (s, 8-C). These spectral data are consistent with the published values for hispidol B (2) [14]. Identity was established through diit comparison with an au~entic sample. REFERENCES Acetylation of 2. Compound 2 (5 mg) was dissolved in Aczctpyridine (1: 1, 1 ml) at room temp. overnight, dried in Wani, M. C.. Taylor, H. L., Thompson, J. B. and Wall, M. E. uacuo and chromatographed over silica gel eluting with CHCI, to (1978) Lloydia 41,578. afford a crystalline triacetate 4 from EtzG-hexane, mp 146-148”: Arisawa M., Kinghorn, A. D., Cordell, 6. A. and ‘H NMR (CDCl,): 60.76 (3H, s, 19-H,), 0.80 (3H, s, 18-H,), 0.85 Farnsworth, N. R. (1983) J. Nor. Prod. 46,218. (3H,s,U)-H,),0.96(3H,s,29-H,),0.97(3H,s,28-H,),0.98 (3H,d, Polonsky. J., Gallas. J.. Varenne, J., Range, T., Pascard, C., J = 6.8 Hx 21-H,), 1.19 (3H.s. 2&H,), 1.25 (3H.s, 27-H,), 2.06, Jaquemin, H. and More& C. (1982) Tetruhedron Letters 23. 869. 2.07 and 2.20 (each, 3H, s, OAc), 4.53 (1H. m, 3a-H), 4.89 (IH, m, 24-H), 5.23 ( 1H. br s, 7-H) and 5.23 (1H, br s, 7-H) and 5.42 (1H, m, Arisawa. M., Fujita A., Morita. N., Kinghorn, A. D.. Cordell, 23Hf; MS mjz: 602 CM]‘. 569, 542, 527, 509,484,467,449;425, G. A. and Farnsworth, N. R. (1985) Plonta Med. SO. 348. 407, 389, 369, 353, 335 and 309. Arisawa M., Kinghorn, A. D.. Cordell, G. A. and Farnsworth. N. R. (1983) J. Nut. Prod. 46,222. Reduction of 1 with LAH. To a THF soln of 1 (4 mg), LAH Arisawa M.. Handa S. S., McPherson, D. D., Lankin, D. C., (1 mg) was added and the mixture reftuxed for 1 hr. The reaction Cordell, G. A., Fang, H. H. S. and Farnsworth, N. R. (1984). mixture was worked up as usual to afford a reduction product (3, J. Nat. Prod. 47.300. 1.3 mg) as colourless needles, mp 142-144” (Me&O). ‘H NMR
Triterpenes from Simaba multiflora 7. Arigoni. D.. Barton. D. H. R., Corey, E. J.. Jeger, 0.. Caghotim, L., Dev. S., Ferrini, P. G.. Glazier. E. R., Melera, A., Pradhan. S. K.. Schaffner, K., Stemhell, S., Templeton, J. F. and Tobinaga. S. (1960) Experientio 16,41. 8. Bevan. C. W. L.. Ekong D. E. U.. HaBall, T. G. and Taft, P. (1967) 1. Chem. Sot. C 820. 9. Merrien, A. and Polonsky, J. (1971) Chem. Commun. 261. IO. Polonsky, J.. Baskevitch-Varon. Z. and Das. B. C. (1976) Phytochemisrry 15, 337. I I. Polonsky. J., Varon, Z., Rabanal, R. M. and Jacquemin, H. (1977) Isr. J. Chem. 16, 16. 12. Sherman, M. M., Borris. R. P.. Ogura, M.. Cordell, G. A. and Farnsworth, N. R. (1980) Phytochemisrry 19, 1499.
3303
13. Chan, W. R.. Taylor, D. D. and Yee, T. (1970) 1. C/mm. Sot. C3ll. 14. Jolad, S. D., Hoffmann. J. J. and Cole, J. R. (1981) J. Org. Chem. 46.4085. IS. Djxassi, C. and Geller, L. E. (1959) J. Am. Chem. Sot. 81, 2789. 16. Ohmoto. T., Tanaka, R. and Nikaido, T. (1976)Chem. Pharm. Bull. 24, 1532. 17. Jurd. L.. Stevens. K. and Manners, G. (1972) Phytochemistry 11. 2535. 18. Gilmore. C.. J. (1984) J. Appl. Cryst. 17, 42. 19. Sheldrick. G. M. (1976) A Program for Crystal Structure Determinerion. University of Cambridge.
TRITERPENES
0
003 I -9422/87 f3.00 + 0.00 1987 Pmgamon Journala Ltd.
FROM SZMABA MULTZFLORA
MUNEHISA ARISAWA, AKIO FUJITA, NAOKATA MORITA, PHILIP J. Cox; CORDELL~ Facultyof Phamuuxutical Sciences, Toyama Medical and P -tical of Pharmacy, Robert Gordon’s Institute of Technology, Schoolhill,
R. ALAN How&t
and GEOFFREY
A.
University,2630 SugitaniToyema 930-01, Japan;*School
Aberdeen, AR9 IFR, Scotland; tChemistry Department, University of Aberdeen, M&on Walk, Old Aberdeen, AB9 2UE. Scotland; $Program for Collaborative Restarch in the Pharmaceutical Sciences, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois 60612, U.S.A. (Revised receiwd 6 April 1987)
Key Word Imdcx-Simaba mulfifloro; Simaroubaaae; canthin-6-onc; 8-0-methylretusin; vanillic acid; hispidol B; 3$23R,25-trihydroxytirucall-7un-24-one; X-ray analysis.
Abstract-A new triterpene has been isolated from Sin&a multijba (Simaroubaceae) and determined to have the structure 3S23R.25-trihydroxytirucall-7-en-24-one (1). Hispidol B was confirmed to be 3S,23S,24R,25tetrahydroxytirucall-7-ene (2) by X-ray crystallographic analysis. Canthindane, 8-O-methyl retusin and vanillic acid were also obtained in the course of this work.
lN’fRODUCTlON
Sima6a multifira A. Juss. (Simaroubaceae) has previously afforded several quassinoids [l-4], canthin&ones [4,5], canthin-2,6diones [S], coumarins [4,5], a coumarinolignan [6], two phenolic compounds [4] and j?-sitosterol+~ glucoside [2]. The quassinoids are of interest because of their anticancer activity, and biogenetically the skeleton is derived from a tin&l-7ene derivative by way of an apoeuphol rearrangement [7,8]. However, tirucall-7cne derivatives have been obtained only occasionally from Simaroubaceous plants [9-121 and we report here on the isolation and structure determination of a new member of this series.
these two protons in 4 and the negative CD curve for 1, the stereochemistry of C-23 [15] was assigned as R. Consequently, the structure of 1 was assigned as 3&23R,25-trihydroxytirucall-7-eu-24-one. Compound 2, mp 252-254”, was found to be the known compound hispidol B, whose structure was previously assigned in ref. [14]. In order to thoroughly establish the stereochemistry in the side chain. X-Ray crystallographic analysis of this compound was performed, through which its structure was wniirmed lo be 3S,23&24R,25tetrahydroxytirucall-7-e. Canthindane [ 161 and 8-Omethylretusin [17] were identified by comparison with spectral data. Vanillic acid was identified by direct wmparison with an authentic sample.
RESULTS AND DISCUSSION
In a continuing search for anticancer agents from the wood of Simaba multipora [2, 4-61, a new triterpene was isolated and determined to be (3&23R,25)-hihydroxytirucall-7en-24-one (1). Hispidol B (2) was also obtained and confirmed lo be 3S,23&24R,25tetrahydroxytirucall-7-ene by X-ray analysis. Compound 1, mp 208-213”. molecular formula C,,H,,O, by high resolution mass spectrometry. exhibited IR absorption bands at 3500, 3250 (OH) and 169Ocm-’ (GO). The ‘H NMR spectrum of 1 indicated the presence of seven tertiary methyl groups, one secondary methyl group, two multiplets for oxymethine protons and one o1efGc proton. These signals were regarded as being characteristic of the tirucall-7cne system having a 3-hydroxy group [12-141. A singlet for two methyl groups shifted downfield to 6 1.30 (6H) suggesting attachment to an oxygenated carbon, i.e. C-25. An oxymethine proton was shifted to 63.83 indicating the presence of a vicinal diol, and an ion at m/z 459 [M - Me] + for loss of the C14 methyl supports the assignment of a double bond to C7. LAH reduction of 1 afforded 3, the C-23, C-24 isomer of 2. in which both the 23-H and 24-H were shifted substantially upfield. From the coupling constant between 3301
R’
b
R’
R’
0
H
OH
H
/OH ‘I/,~ <”
“OH /OAc
H
OH H
0:
H
““H
OAc
H
AC
/OAc ” “H
H
OAc
AC
M. ARISAWAet al.
3302 EXPERIMENTAL
(CDCI,): 60.75 (3H, s, 19-H,), 0.83 (3H, s, 18-H,), 0.86 (3H, s, 3@ H,), 0.97 (6H, s, 28 and 29-H& 1.03 (3H, d, J = 5.8 Hz, 21-H,), Mps were determined by means of a hot plate and are uncorr. 1.30 (6H. z, 26 and 27-H,), 3.25 (1H. dd, J = 5.3,10.9 Hz, 3a-H), ‘HNMR~od”CNMRspcctrowercr~rdedat200MHzmd 3.28 (Hi, d, J = 5.7 Hz, 24-H), 3.83 (lH, ddd, J = 3.2,5.7,5.7 Hz, 50.3 MHz, respectively (TM& int. standard, chemical shifts: 6). 23-H) and 5.26 (lH,dd. J = 0.8.2.2 & 7-Hk MS m/z: 476 [hi) l, Prelimialuy p?oces.wng end isolation. Details coaceraing the 461, 443, 425, 406, 4OQ 386, 371, 353. 327 and 309; Mass collection, identification and prelimimuy fractionation of the measurement,obs.: 476.3858. c&. for Cz0HSz04, 476.3863. wood of Sin&a muIripor0 A. Juss. (Simaroubaaac) have been Aceryfation of3. The reduction product 3 (1 mg) was dissolved in described previously [2,4-6]. In a oontinution of the previous A@-pyridine (1: 1, 0.3 ml) at room temp. overnight, dried In study [6], the mother liquor after the isolation of emodin was wrcuo and chromatographed over silica gel to alford a triacemtc 5, rechromatogtapbed on silica gel eluting with hexane-EtOAc (15) mp 152-155” (MeGH). ‘H NMR (CDC&): 60.76 (3H, s, 19-HJ). to afford I-0-methylretusin (10 mg), and anthiod-one (3 mg). 0.79 (3H,s, la-H,),0.85 (3H,s, 30-H&0.93 (3H, s, 29-H,),0.94 Droplet countercurrent chromatography (DCCC) of the com(3H,d,J = 5.1 Hx,21-H,),O.% (3H,s.28-H,), 1.20(3H,s,26or bined fractions 11 and 12, using a biphasic mixture of 27-H,), 1.25(3H,s,27or2~H,A4.51 (lH,dd,J = 5.3.9.3 Hx,3aCHClx-CeHe-MeGH-Hz0 (3: 1: 5:2) afforded t&in, 8- H), 4.93 (1H. d, J = 2.2 Hx 7-H), 5.23 (1H. m, .I = 3.6 Hx, 23-H) hydroxycanthi&-one and simalikalactone D as previously and 5.24 (lH, m, 24Hk MS m/z: 602 [Ml’, 587,569, 542,527. reported [6], and hispidol B (2 12mg), 3S,23R,25- 509,484,482.467,449.425,4C9,407.389.369,353,325 and 309. trihydroxytirucall-?c-24-one (I, 5 mg) and vanillic acid (6 mg). Crysrul data of 2. C,,H,zO., M = 476.4. ~horhombi~ u 3S~3R,25-Trih~~oxytdroxytinccaif-7-en-24-one (1). Recrystaliixed = 10.605(39), b = 14.455(75), c = 18.203(31) A, U = 2790.4A3, from Me&O gave colourless needles, mp 208-213”. CD: [6 Jz9, Z = 4, 0, = 1.13 8ctK3, F(OO0)= 1056, MO-K. radiation, i - 3330” (CHCl,k IR vzcm -I: 3500, 3250. 2950, 2910, 1690, = 0.7108 A p = OAOcm-‘. Space group F&2,2,. 1460,1450,1380,1360and 1060; ‘H NMR (CD&): 60.83 (3H. s, Crystalfagrophic analysis of 2. The X-ray intensities were 18-H,), 1.01 (6H. s, 19-Ha and 30-H& 1.03 (3H. d. J = 6.5 Hr, collated on a Nicolet P3 automated diffractometer using mono21-H,), 1.05 (3H, s, 29-H,), 1.12 (3H. s, 28-H,), 1.30 (6H, s, 26-H, chromatixul MoK, radiation. Integrated relative intensities for and27-Hxk2.69 (lH,m,22-Hk3.28 (lH,m,3-Hk3.83 (lH,m,231375 independent reflexions with 20 < 40” were measured as BH) and 5.32 (1H. m. 7-H); MS m/z: 474 [Ml’. 459 [M - Me]‘, 20scans; all reflexions were used in subsequent calculations. The 441 [M - Me-HzO]+, 423 [M - Me- 2HzO]‘, 369, 351, 325. structure was elucidated using MITHRIL [la] and the Hatoms 271,189,121,72and 55. Mass measurement, obs.: 474.3697. a&. were observed in electron density mapscalculated at intermediate for Cx,H,eO,, 474.3706. stages of structure refinement. The co-ordinates for allatoms and Hizpidol B (2). Recrystallimd from MeGH gave colourless anisotropic thermal parameters for the non-hydrogen atoms needles, mp 252-254”. IR$&cm-i: 3340, 2950. 1450, 1370, were varied in least-squares calculations using SHELX [ 191.The 1160,1070,1030,990,825 and 795; ‘H NMR (pyridine-d,): 60.83 C-H and GH bonds were constrained to be qua1 to 1.00 A and (3H,~l8-H~~O.~(3H,~l9-H~~l~O2{3H,s~~H~~l.l2(3H,~ the hydroxy, methyl and remaining hydrogen were given 29-H,), 1.14 (3H.4 .J = 6.8 Iix, 21-H,), 1.18 (3H, s, 28-H,), 1.62 common temperature factors during refinement. Unit weights (3H, s, 26-H,), 1.64 (3H,s, 27-H,), 1.88 (2H,m, 6-H,), 2.35 (lH, were employed and refinement converged at R 7.2%. Final dd, J = 9.5, 12 Hz, 22-H), 3.49 (lH, dd, J = 8.0 Hx, 31x-H),3.68 positional parameters are listed in Table 1, bond lengths in (lH,br~2~HA4.~(lH,~~ = 4.~9.5~2~H~S.~(lH,~7Table 2, valency angles in Table 3 and torsion angles in Table 4. H),5.53,5.74,5.89and6.08 (each, lH,m,excbangeablewith D,O, Thermal parameters are listed in a Supplementary Publication. OHkMSm/z:476[M]+,461[M-Me]+.458[M-H,0]*,433 [M -Me - HzO]+, 425 [M - CH, -2HzO]+. 407, 400. 385. Acknowledgements--This work was supported, in part, by 372, 353 and 327. Mass measurement, ohs.: 476.3838. talc. for grant CA 20164 from the Division of Cancer Treatment, CxOH>zO*, 476.3866, “CNMR (pyridine-d,): 13.38 (q, 18-Q National Cancer Institute. The authors would also like to thank 15.53 (q, 29-Q 18.37 (t, 11-C), 19.46 (q, 19-C), 24.35 (t, 6-Q 27.04 Professor J. R. Cole at the University of Arizona. for a gift of an (q, 26-Q 27.43 (q, 3&C), 27.66 (q, 27-C), 28.24 (q, 28-C), 2836 (t. authentic sample of hispidol B (2) and Mr M. Morikoshi. 16-Q 28.72 (t, 2-Q 34.13 (t, 12-Q 34.26 (t. 15-Q 34.26 (d, 20-Q Toyama Medical and Pharmaceutical University, for measure35.12 (s, l&C), 37.57 (t, l-C), 39.44 (s, 4-C), 42.03 (t, 22-C). 43.77 (s, ment of the 13CNMR and mass spectra. 13-Q 4925 (4 9-C) 51.07 (d, 25-Q 51.43 (s, M-CA 54.28 (4 17cX 69.29 (d, 23-C), 73.69 (s, 5-C), 76.50 (d, 24-Q 78.20 (d, 3-Q 118.32 (d, 7-C)and 145.95 (s, 8-C). These spectral data are consistent with the published values for hispidol B (2) [14]. Identity was established through diit comparison with an au~entic sample. REFERENCES Acetylation of 2. Compound 2 (5 mg) was dissolved in Aczctpyridine (1: 1, 1 ml) at room temp. overnight, dried in Wani, M. C.. Taylor, H. L., Thompson, J. B. and Wall, M. E. uacuo and chromatographed over silica gel eluting with CHCI, to (1978) Lloydia 41,578. afford a crystalline triacetate 4 from EtzG-hexane, mp 146-148”: Arisawa M., Kinghorn, A. D., Cordell, 6. A. and ‘H NMR (CDCl,): 60.76 (3H, s, 19-H,), 0.80 (3H, s, 18-H,), 0.85 Farnsworth, N. R. (1983) J. Nor. Prod. 46,218. (3H,s,U)-H,),0.96(3H,s,29-H,),0.97(3H,s,28-H,),0.98 (3H,d, Polonsky. J., Gallas. J.. Varenne, J., Range, T., Pascard, C., J = 6.8 Hx 21-H,), 1.19 (3H.s. 2&H,), 1.25 (3H.s, 27-H,), 2.06, Jaquemin, H. and More& C. (1982) Tetruhedron Letters 23. 869. 2.07 and 2.20 (each, 3H, s, OAc), 4.53 (1H. m, 3a-H), 4.89 (IH, m, 24-H), 5.23 ( 1H. br s, 7-H) and 5.23 (1H, br s, 7-H) and 5.42 (1H, m, Arisawa. M., Fujita A., Morita. N., Kinghorn, A. D.. Cordell, 23Hf; MS mjz: 602 CM]‘. 569, 542, 527, 509,484,467,449;425, G. A. and Farnsworth, N. R. (1985) Plonta Med. SO. 348. 407, 389, 369, 353, 335 and 309. Arisawa M., Kinghorn, A. D.. Cordell, G. A. and Farnsworth. N. R. (1983) J. Nut. Prod. 46,222. Reduction of 1 with LAH. To a THF soln of 1 (4 mg), LAH Arisawa M.. Handa S. S., McPherson, D. D., Lankin, D. C., (1 mg) was added and the mixture reftuxed for 1 hr. 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13. Chan, W. R.. Taylor, D. D. and Yee, T. (1970) 1. C/mm. Sot. C3ll. 14. Jolad, S. D., Hoffmann. J. J. and Cole, J. R. (1981) J. Org. Chem. 46.4085. IS. Djxassi, C. and Geller, L. E. (1959) J. Am. Chem. Sot. 81, 2789. 16. Ohmoto. T., Tanaka, R. and Nikaido, T. (1976)Chem. Pharm. Bull. 24, 1532. 17. Jurd. L.. Stevens. K. and Manners, G. (1972) Phytochemistry 11. 2535. 18. Gilmore. C.. J. (1984) J. Appl. Cryst. 17, 42. 19. Sheldrick. G. M. (1976) A Program for Crystal Structure Determinerion. University of Cambridge.