Alkaloids from Brunsvigia orientalis

Pergamon Phytocheraistry, Vol. 43, No. 6, pp, 1379-1384, 1996 PII: S0031-9422(96)00506-7 Copyright © 1996 Elsevier Science Ltd Printed in Great Britain. All rights reserved 0031-9422/96 $15.00 + 0.00 ALKALOIDS FROM BRUNSVIGIA ORIENTALIS FRANCESCVILADOMAT,GIOVANNAR. ALMANZA,CARLESCODINA,JAUMEBASTIDA,* WILLIAM E. CAMPBELL'~and SHAHEEDMATHEEt Departament de Productes Naturals, Facultat de Farmhcia, Universitat de Barcelona, 08028 Barcelona, Catalonia, Spain; +Chemistry Department, University of Cape Town, Rondebosch 7700, South Africa (Received 27March 1996) Key WordIndex--Brunsvigia orientalis; Amaryllidaceae; bulbs; alkaloids; lycorine; crinine; buphanisine; buphanidrine; epibuphanisine; undulatine; crinamidine; crinamine; 6-hydroxycrinamine; l-epibowdensine; 1-epidemethoxybowdensine; 1-epideacetylbowdensine. Abstract--Twelve alkaloids have been isolated from bulbs of Brunsvigia orientalis. 1-Epibowdensine, lepidemethoxybowdensine and 1-epidecacetylbowdensine are reported here for the first time. The structure and stereochemistry of these new alkaloids have been determined by physical and spectroscopic methods. The ~H and 13C NMR spectra of 6-hydroxycrinamine (both epimers) and crinamidine were completely assigned by means of 2D NMR techniques. Copyright © 1996 Elsevier Science Ltd INTRODUCTION Brunsvigia orientalis is an endemic Amaryllidaceae species from southern Africa, widely distributed in the southern and southwestern Cape [1]. In the present study, we report the isolation and characterization of 12 alkaloids from the bulbs of this hitherto unstudied plant species. Undulatine (2), crinamine (4) and the new alkaloid, 1-epibowdensine (5), were found to be the principal constituents. According to the literature, compound 2 was described as an active antineoplastic agent from Amao'llis belladonna bulbs [2]. In turn, compound 4 was the principal antibacterial constituent of the bulbs of Crinum jagus [3], possessing strong cytotoxic and moderate antimalarial activities [4]. Compound 5, as well as 1-epidemethoxybowdensine (6) and l-epideacetylbowdensine (7), were the first natural /35,10b-ethanophenanthridine alkaloids having the C-1 substituent in the equatorial orientation. It is noteworthy that, just as in B. josephinae [5, 6], the majority of alkaloids from B. orientalis belong to the 5,10bethanophenanthridines; many of them have a methoxyl group at the C-7 position. RESULTS AND DISCUSSION The absolute configurations of alkaloids with a 5,10b-ethano bridge were determined from circular dichroism (CD) curves. In this way, the molecular ellipticity of crinamidine (1), 2, 5-7, crinine, *Author to whom correspondence should be addressed. buphanisine and buphanidrine showed CD curves which were similar qualitatively to those of/3-5, 10bethanophenanthridine alkaloids with a maximum around 250 nm and, in turn, the CD curves of 6hydroxycrinamine (3a and 3b), 4 and epibuphanisine were similar to those of a-5,10b-ethanophenanthridines with a minimum around 250 nm [7, 8]. Compound 1, C17HI9NO5, characterized as crinamidine, gave an El mass spectrum with a base peak at m/z [M-29] + and exhibited the typical fragmentation pattern for structures with an epoxide ring [9]. The JH NMR spectrum was completely assigned, providing additional information with respect to a previously reported paper [10]. The spectrum was similar to that of 2 [6], apart from the absence of the singlet attributable to a methoxyl group. The small coupling constants between H-1 and H-2 (J = 4.0 Hz), H-2 and H-3 (J = 2.5 Hz) and between H-3 and H-4/3 (J = 3.0 Hz), together with the additional long-range coupling of H-2 with H-4a (W-mechanism), allowed us to assign the configurations of the epoxide ring and the hydroxyl group at the C-3 position. The aromatic singlet proton was assigned to the C-10 position, because of the three-bond HMBC correlations with C-6a, as well as with C-10b and C-8, and this was corroborated by the spatial proximity (ROESY [11] experiment) between H-10 and H-1. Thus, it could be inferred that the position of the methoxyl group was at C-7. The ~3C spectrum (Table 1) is reported for the first time and all resonances have been unambiguously confirmed by means of HMQC [12] and HMBC [13] experiments. The pronounced highfield shift of the C-1 and C-2, signals, with respect to the corresponding 1379 1380 F. VILADOMATet al. I~,. 2 ,oR 1 lO o,.~ 9.,,,"~ °%" 12 6 OMe 1: R=H 2: R=Me 2 , 1o .OMe I ..'k---l- OH 9 4.\,,,"J" 3a: 3b: 4: RI=H, R2=OH RI=OH, R2=H RI=R2=H OR2 2 171Oy ~ 3 R3 5: RI=R2=Ac, R3=OMe 6: RI=R2=Ac, R3=H 7: Rt=R2=H, R3=OMe signals in the 1,2-unsaturated alkaloids in this series, was consistent with an 1,2-oxiran ring; substitution of the methoxyl group at the C-3 position by an hydroxyl group induced a pronounced shielding effect on C-3 and a deshielding effect on the/~-carbons, C-2 and C-4, with respect to 2 [6]. 6-Hydroxycrinamine, C]7HIgNOs, recrystallized as needles from acetone was homogeneous chromatographically but the signals in both 'H and '3C NMR spectra were rather complex, suggesting that it was a 2:1 mixture of two epimers (3a and 3b) in the C-6 position. The E1 mass spectrum showed a typical fragmentation pattern of 1,2-unsaturated alkaloids of the crinine series bearing a hydroxyl substituent at C-I 1 [14]. In this group, the loss of methanol (peak at m/z 285) is favoured when both the ethano bridge and the methoxyl group at the C-3 position are on the same side of the molecule and, in the case of derivatives bearing a 6-hydroxyl substituent, ion [M-methanol] + very easily loses a hydroxyl radical leading to the base peak (m/z 268), which is consistent with 6-hydroxycrinamine data [14]. The 'H NMR spectrum reported in the literature is incomplete [15] and all 'H resonances of both epimers were therefore unambiguously assigned (Table 2). In both, the small coupling constant between H-2 and H-3, the large one between H-4a and H-3, and finally the NOE contour correlation between H-3 and H-4a, were also indicative of a cis-relationship between the C-3 pseudoequatorial substituent and the 5,10b-ethano bridge. The deshielding effect on H-11, in relation to the reported data for alkaloids with no bridge substituent in this series [16, 17], as well as the NOE effect between H-10 and H-11 and the long range W-coupling between H-11 and H-4a, were consistent with a C-11 hydroxyl substituent at the exo-position. The epimer 3a showed the benzylic proton H-6a as a singlet at 6 5.01 and spatial proximity between H-6 and H-12 endo was observed. In the 3b spectrum, the proton H-6fl was observed at lower fields (6 5.59) and a NOE contour correlation between H-6fl and H-4a was established, confirming the assignation of the hydroxyl group at C-6 in both epimers. The '3C assignments (Table 1), reported for the first time, were confirmed by HMQC and HMBC experiments. The pronounced deshielding effect on C-11 and C-6 of both epimers, observed as doublets, was also consistent with the presence of hydroxyl substituents. In both structures, 3a and 3b, the carbon singlet C-9 was assigned at lower fields than C-8 because of its three-bond correlation with the methine proton H-7. The quaternary carbons C-6a and C-10a were ascribed by means of their correlations with the methine protons H-10 and H-7, respectively. Finally, the singlets at 6 50.4 and 50.8 were assigned to C-10b of 3a and 3b, respectiely, taking into account their three-bond connectivities with H-2, H-4 and H-10. Compound 5, C21H25HO 7, as well as its 7-demethoxyl derivative, compound 6, C2oHz3NOr, were isolated for the first time from a natural source. Their EI mass spectra displayed a similar fragmentation pattern, with a parent peak at m/z 403 and 373, respectively, and important fragments at m/z [M-59] +, [M-119] +, [M-131] +, [M-149] + and [M-171] +, which are characteristic of 1,2-disubstituted crinamine alkaloids [18]. Compound 5 showed an additional ion at m/z 314, associated with the loss of both the methoxyl and one acetoxy group. The peaks at m/z [M-59] ÷ and [M-II9] + were in agreement with the loss of one or two vicinal acetoxy groups. Their JH NMR spectra (Table 3) were very close and only the absence of a methoxyl group in 6 was noteworthy. The methoxyl group of $ was assigned to the C-7 position because of 1381 Alkaloids from Brunsvigia orientalis Table 1. ~3C NMR chemical shift assignments of compounds 1 - 7 C 1 2 3a 3b 4 5 6 7 1 2 3 4 4a 6 6a 7 8 9 10 10a 10b 11 12 -OCH203-OMe 7-OMe l-OAc 53.8d 56.4 d 65.5 d 29.7 t 61.0 d 58.6 t 117.6 s 141.1 s 133.4 s 148.1 s 96.4 d 138.7 s 41.6 s 39.2 t 52.5 t 100.7 t 53.9d 55.1 d 74.8 d 25.2 t 61.2 d 58.6 t 117.8 s 140.9 s 133.3 s 147.9 s 96.3 d 138.9 s 41.4 s 39.2 t 52.5 t 100.5 t 57.5 q 59.0 q 136.4d 123.0 d 75.9 d 29.4 t 59.5 d 88.0 d 127.3 s 109.5 d 146.5 s 147.8 s 102.8 d 135.8 s 50.4 s 78.1 d 57.7 t 101.1 t 55.9 q 136.2d 123.2 d 75.6 d 29.4 t 64.8 d 85.5 d 128.8 s 108.3 d 146.7 s 147.5 s 102.7 d 134.6 s 50.8 s 79.0 d 51.8 t 101.1 t 55.9 q 136.8 d 123.6 d 76.0 d 30.1 t 66.1 d 63.5 t 126.6 s 106.8 d 146.1 s 146.4 s 103.1 d 135.5 s 50.2 s 80.0 d 61.2 t 100.7 t 55.6 q 74.0d 68.2 d 26.3 t 21.0 t 68.1 d 58.3 t 116.9 s 140.3 s 133.4 s 148.0 s 97.3 d 140.9 s 47.0 s 37.2 t 52.2 t 100.5 t 74.0d 68.3 d 26.4 t 21.3 t 68.5 d 62.2 t 125.8 s 106.4 d 146.0 s 146.2 s 103.6 d 140.0 s 47.2 s 37.4 t 52.2 t 100.8 t 73.0d 69.7 d 28.9 t 20.4 t 67.6 d 58.2 t 116.5 s 140.0 s 133.3 s 148.0 s 99.5 d 142.1 s 49.3 s 36.4 t 51.7 t 100.4 t 59.0 q 21.2 q 170.0 s 21.2 q 170.4 s 21.1 q 170.0 s 21.2 q 170.4 s 59.1 q 2-OAc the t h r e e - b o n d H M B C correlations b e t w e e n H - 1 0 a n d C-6a, as well as with both C - 1 0 b and C-8, a n d b e t w e e n the H-6 protons a n d C-7, a n d b e c a u s e o f the o b s e r v e d N O E b e t w e e n H - 1 0 a n d H-1. C o m p o u n d 6 s h o w e d two para-positioned aryl protons, w h i c h were consistent with their multiplicity. Both alkaloids h a v e two a c e t o x y g r o u p s at C - I a n d C-2; a d e s h i e l d i n g effect on H-1 a n d H-2 w a s observed. A R O E S Y e x p e r i m e n t allowed u s to establish the axial orientation o f H-1 by spatial p r o x i m i t y with H-10, as well as with H-3ax. T h e m a g n i t u d e o f the coupling c o n s t a n t b e t w e e n H-1 and H - 2 a n d b e t w e e n H-2 and H - 3 a x led us to a s s i g n the equatorial disposition for H-2. C o n s e q u e n t l y , the acetoxy substituents on C-1 and C-2 s h o u l d be a s s i g n e d to the equatorial and axial disposition, respectively. T h e large vicinal coupling 59.0 q c o n s t a n t s b e t w e e n H - 4 a x a n d H - 4 a (J = ca 12.0 Hz) a n d b e t w e e n H - 4 a x and H - 3 a x ( J = 13.5 Hz) denoted their trans-diaxial relationship, w h i c h w a s consistent with the N O E contour correlations b e t w e e n H - 4 a a n d H - 3 a x and b e t w e e n H - 4 a x a n d H-12exo. The assignm e n t s o f the H - 6 and H - 1 2 protons were supported by the N O E effect b e t w e e n H - 1 2 e n d o and H-6fl, as well as b e t w e e n H - 6 a and H-4a; additionally, both H - 6 a and H-12exo protons were a s s i g n e d at lower fields d u e to their cis-relation with the nitrogen lone pair [19]. T h e 13C N M R spectra o f 5 a n d 6 were a s s i g n e d taking into a c c o u n t the H M Q C a n d H M B C c o n n e c tivities (Tables 1 a n d 4). A t lower fields six (6) or s e v e n (5) carbon singlets for the acetoxy carbonyl groups a n d the quaternary c a r b o n s o f the aromatic ring were observed. T h e carbon singlet at 8 140.3 o f 5 w a s Table 2. ~HNMR and ROESY data for compounds 3a and 3b (J given in Hz in parentheses) H 3a H NMR 1 2 3 4a 413 4a 6 7 10 11 6.23 d (10.5) 6.19 dd (10.5, 2.0) 4.02 ddd (9.0, 6.5, 2.0) 2.11 ddd (13.0, 12.5, 9.1) 2.08 ddd (12.5, 6.5, 5.0) 3.73 ddd (13.0, 5.0, 1.0) 5.01 s 6.80 s 6.74 s 3.90 ddd (6.5, 3.0, 1.0) 12endo 3.35 dd (14.0, 6.5) 12exo 3.30 dd (14.0, 3.0) -OCH20- 5.89 d-5,91 d (1.5) 3-OMe 3.37 s 3b ROESY ~H NMR ROESY H-2, H-10, H-11 H-I, H-3 H-4a, H-4/3, H-2 6.21 d (10.5) 6.17 dd (10.5, 1.5) 3.96 ddd (10.5, 6.0, 1.5) 2.26 ddd (13.5, 12.5, 10.0) 2.16 ddd (12.5, 6.0, 5.0) 3.41 ddd (13.0, 5.0,1.0) 5.59s 6.96 s 6.72 s 3.87ddd(7.0,2.5, 1.0) 4.19 dd (14.0, 7.0) 3.01 dd (14.0, 2.5) 5.88 d-5.90 d (1.5) 3.38s H-2, H-10 H-I, H-3 H-4a, H-4/3, H-2 H-4fl, H-12exo H-4ot, H-3, H-4a H-3, H-4/3 H-12endo, H-7 H-6 H-I, H-I 1 H-l,H-lO, H-12endo H-6, H-12exo, H-11 H-4ct, H-12endo H-4/3,H-12exo H-4a, H-3, H-4a H-3, H-4fl, H-6 H-4a, H-7 H-6 H-I H- 12endo H-12exo, H-11 H-4a, H- 12endo 1382 F. VILADOMATet al. Table 3. JH NMR data for compounds 5-7 (J given in Hz in parentheses) H 5 6 7 l 2 3eq 3ax 4eq 4ax 4a 6or 6fl 7 10 1lendo 1 lexo 12endo 12exo -OCH20I-OAc 2-OAc 7-OMe 5.30 d (4.0) 5.55 ddd (4.0, 3.5, 2.0) 1.92 dddd (14.0, 3.5, 3.0, 2.5) 1.57 dddd (14.0, 13.5, 3.5, 2.0) 1.60 dddd ( 14.0, 5.5, 3.5, 3.0) 1.67 dddd (14.0, 13.5, 11.5, 2.5) 3.01 dd (11.5, 5.5) 4.16d d (17.5) 3.74 d (17.5) 5.32 d (4.5) 5.56 ddd (4.5, 3.5, 2.5) 1.93 dddd (14.0, 3.5, 3.0, 3.0) 1.56 dddd (14.0, 13.5, 3.5, 2.5) 1.60 dddd ( 14.0, 5.5, 3.5, 3.0) 1.68 dddd (14.0, 13.5, 12.0, 3.0) 3.06 dd (12.0, 5.5) 4.33 d (17.0) 3.74 d (17.0) 6.42 s 6.43 s 2.03 ddd (12.5, 9.0, 4.5) 2.75 ddd (12.5, 10.5, 5.5) 2.82 dd (12.5, 9.0, 5.5) 3.41 ddd (12.5, 10.5, 4.5) 5.84 d-5.85 d (1.5) 2.09 s 2.09 s 4.08 d (4.5) 4.17 ddd (4.5, 3.5, 2.5) 2.06 dddd (14.0, 3.5, 3.5, 3.0) 1.56 dddd (14.0, 12.5, 3.5, 2.5) 1.59 dddd ( 14.0, 5.0, 3.5, 3.0) 1.79 dddd (14.0, 12.5, 11.5, 3.5) 2.95 dd (11.5, 5.0) 4.22 d (17.5) 3.80 d (17.5) 6.16 s 2.01 ddd(12.5, 9.0, 4.5) 2.73 ddd (12.5, 10.5, 5.5) 2.80 ddd (12.5, 9.0, 5.5) 3.40 ddd (12.5, 10.5, 4.5) 5.80 d-5.82 d ( 1.5) 2.08 s 2.08 s 3.95 s 7.24 s 2.01 ddd (12.0, 8.5, 4.5) 2.78 ddd (12.0, 9.5, 6.0) 2.80 ddd ( 12.5, 8.5, 6.0) 3.42 ddd (12.5, 9.5, 4.5) 5.88 d-5.90 d (1.5) 4.00 s assigned to C-7 because of its three-bond correlation with the methoxyl group and both H-6 protons. Moreover, the additional methoxyl group strongly influenced the signals of C-6a and C-8. In contrast, the C-7 doublet of 6 was observed in the characteristic shift range (8 106.4). The rest of the signals were very close for both 5 and 6. Thus, the carbon singlets of the acetoxy carbonyl groups (ca ~ 170) were assigned taking into account the three-bond connectivities with the H-1 or H-2 protons. The quaternary carbons of the aromatic ring, as well as C-10b, were easily assigned by means of long-range correlations. The other new alkaloid (7) was isolated as a crystalline white compound. The E1 mass spectrum showed a [M] + at m / z 319, which analysed for C17H21NO~ and exhibited only a few prominent peaks, one being that at m / z 232, being consistent with a 1,2-disubstituted crinane alkaloid [10, 18]. The IR spectrum showed an intense absorption band at 3500-3300 cm -~, characteristic of a hydroxyl group but no carbonyl absorption was observed. Its ~H N M R spectrum (Table 3), recorded in CDC13, was similar to that of 5, only the absence of the singlets attributable to the acetoxy groups was noteworthy. A ROESY experiment was used principally to afford information about the relative spatial distances of protons, allowing us to establish the axial orientation of H-1 by spatial proximity to H-10, H-3ax and H-4a. The small coupling constants of H-2 confirmed its equatorial disposition. All of these data would allow the assignment of the proposed structure, which has the same stereochemistry as the related alkaloid 5. The ~3C assignments (Table l ) were confirmed considering the connectivities obtained from HMCQ and HMBC spectra. Table 4. ROESY and HMBC data for compound 5 H ROESY HMBC 1 2 3eq 3ax 4eq 4ax 4a 6or 6fl l0 1 lendo 1 lexo 12endo 12exo -0CH201-OAc 2-OAc 7-OMe H-2, H-3ax, H-10 H-I, H-3eq, H-3ax H-3ax, H-2, H-4eq, H-4ax H-3eq, H-4a, H-4eq, H-2, H-1 H-3eq, H-3ax, H4ax, H-4a H-3eq, H-4eq, H-12exo H-3ax, H-4eq, H-6a H-4a, H-6fl H-6c~, H- 12endo H-1 H-l lexo, H- 12,endo H-1 lendo, H-12exo H-12exo, H-l lendo, H-6fl H-12endo, H-1 lexo, H-4fl C-I 1, C-4a, C- 10a, CO C-10b C-2 C-12, C- 10a, C-11 C-10a, C-12, C-7 C-10a, C-12, C-7, C-4a C-8, C-6a, C-10b C-10a C-lOa C-lOb C-8, C-9 C-7 Alkaloids from Brunsvigia orientalis EXPERIMENTAL General. Mps are uncorr. IR spectra were measured in KBr discs or as dry films. ElMS at 70 eV. 1H, '3C NMR, DEPT, ~H COSY, HMQC, HMBC and ROESY spectra were recorded in a Varian VXR 500, using the solvent specified and TMS as int. standard. Chemical shifts are reported in 6 units (ppm) and coupling constants (J) in Hz. Silica gel Merck (70-230 mesh) and silica gel SDS chromagel 60 A CC (230-400 mesh) were used for CC and flash CC, respectively. Sephadex LH-20 was used for gel filtration, and silica gel 60 F254 (Merck) for analyt. (0.25 mm) and prep. (1 mm) TLC. Spots on chromatograms were detected under UV (254 nm) and by Dragendorff's reagent. Plant material. Bulbs of B. orientalis (L.) Ait ex Eckl were collected in February 1994 in the southern cape town of Knysna, South Africa. A voucher specimen (Viviers s.n.) has been deposited in the Bolus Herbarium, University of Cape Town, South Africa. Extraction and isolation of alkaloids. Bulbs (5.1 kg) were crushed and macerated with EtOH for 48 hr. The extract was evapd under red. pres., the residue dissolved in H20 and acidified to pH 4. After removing neutral material with Et20, the acidic soln was extracted with CHCI 3 to provide extract A. Basifying the soln to pH 8-9 and extracting it with CHC13 gave extract C. Finally, CHC13-MeOH (3:2) extraction of the basic soln gave extract D. Extracts A, C and D were combined (22.06 g) and subjected to CC on silica gel, eluting with CH2Clz-MeOH (19:1), increasing the gradient for the last steps to (4: 1). Five frs were afforded. Fr. I was subjected to flash CC using a Me2CO-MeOH step gradient; after final purification on Sephadex LH-20, 2 (620 mg) and 5 (460 mg) were isolated. Fr. II was subjected to CC using a CHCI 3MeOH step gradient, followed by further prep. TLC, eluting twice with MeOH and MezCO; after purification on Sephadex LH-20, 5 (62 mg), epibuphanisine (46 mg), buphanidrine (52 mg), 2 (18 mg), 6 (15 mg) and buphanisine (37 mg) were isolated. Compound 4 crystallized directly from fr. III; recrystallization with MeOH afforded 2.12 g. The rest of fr. III was purified in a similar manner to that described for fr. II, and 4 (205 mg), buphanisine (124 mg) and 1 (21 mg) were isolated. Finally, after purification by similar chromatographic processing to that described for fr. II, fr. IV afforded lycorine (87 mg), 7 (29 mg) and 3 (22 mg), and, fr. V, crinine (23 rag). Crinamidine (1). Found: C, 65.05; H, 6.06; N, 4.35. Calc. for C~vH~gNOs; C, 64.35; H, 5.99; N, 4.42%. Mp 215-217 °. [~1~ ~ - 1 0 ° (CHC13; c 0.1). CD [O]253 + 1625, [0]29 o - 1 7 5 . IR Vm,~cm ~: 3400-3200 (-OH), 1498, 1260, (epox.), 1043, 940 (-OCH20-), 805 (epox.). ElMS 70 eV, m/z (rel. int.): 317 [M] + (37), 288 (100), 258 (18), 245 (31), 244 (25), 217 (32), 205 (31), 204 (21), 203 (32), 189 (19), 173 (38), 115 (17), 85 (19), 57 (21), 56 (31). ~H NMR (500 MHz, CDC13): 6 1.56 (1H, ddd, J = 13.5, 12.5, 3.0 Hz, H-4fl), 1.61 (1H, dddd, J = 13.5, 5.5, 2.0, 1.5 Hz, H-4ce), 2.0 (IH, ddd, J = 12.5, 9.0, 5.0 Hz, H-I lendo), 2.37 (IH, ddd, J = 1383 12.5, 10.5, 5.5 Hz, H-11exo), 2.77 (1H, ddd, J = 12.5, 9.0, 5.5 Hz, H-12endo), 3.17 (1H, ddd, J = 12.5, 10.5, 5.0 Hz, H-12exo), 3.17 (1H, dd, J = 12.5, 5.5 Hz, H-4a), 3.26 (1H, ddd, J = 4 . 0 , 2.5, 1.5 Hz, H-2), 3.71 (1H, d, J = 17.5 Hz, H-6fl), 3.75 (1H, d, J = 4 . 0 Hz, H-I), 3.95 (3H, s, 7-OMe), 4.19 (1H, d, J = 17.5 Hz, H-6a), 4.48 (1/4, ddd, J = 3 . 0 , 2.5, 2.0 Hz, H-3), 5.85-5.86 (2H, 2d, J = 1.5 Hz, OCH20), 6.61 (1H, s, H-10). 13C NMR (50 MHz, CDC13): Table 1. 6-Hydroxycrinamine (3a and 3b). Found: C, 63.41; H, 6.07; N, 4.33. Calc. for CITH19NOs: C, 64.35; H, 5.99; N, 4.42%. Mp 150-152 °. [a] 22 + 4 0 ° (CHCI~; c 0.5). CD [O]z56 - 1565, [0]266 +4087. IR Vma~ cm-~: 3420 (-OH), 2926, 1481, 1248, 1038, 933 (-OCH20-). ElMS 70 eV, m/z (rel. int.): 317 [M] ÷ (1), 285 [ M MeOH] ÷ (39), 284 (10), 269 (20), 268 [ M - M e O H OH] + (100), 258 (10), 227 (25), 209 (26). ~H NMR (500 MHz, CDCI3): Table 2; ~3C NMR (50 MHz, CDC13): Table 1. l-Epibowdensine (5). Found: C, 61.55; H, 6.12; N, 3.51. C:lH25NO 7 requires: C, 62.53; H, 6.20; N, 3.47%. Mp 124-126 °. [a]~2 + 4 ° (CHC13; c 1.1). CD [0]256 +2130, [O]286-132. IR Vm~~ cm-~: 2946, 1740 (>C=O), 1617, 1478, 1367, 1245, 1041, 942 (-OCH20-), 751. ElMS 70 eV, m/z (rel. int.): 403 [M] + (100), 344 [ M - OAc] + (81), 314 [ M - OAc - O M e ] ÷ (23), 284 [ M - O A c - H O A c ] ÷ (63), 283 (22), 272 (31), 256 (21), 255 (27), 254 (37), 232 (36), 231 (28), 202 (26). ~H NMR (500 MHz, CDC13): Table 3; ~3C NMR (50 MHz, CDC13): Table 1. l-Epidemethoxybowdensine (6). Found: C, 63.61; H, 6.08; N, 3.81. C2oH23NO 6 requires: C, 64.34; H, 6.17; N, 3.75%. Mp 96-98 °. [a]2D2 + 20 ° (CHC13; c 0.78). CD [O]2~ 6 + 3850, [0]289 - 2 2 5 . IR Vmax cm-~: 2922, 1734 (>C=O), 1478, 1367, 1233, 1036, 942 (-OCH20-), 751. ElMS 70 eV, m/z (rel. int.): 373 [M] ÷ (100), 315 (24), 314 [M - O A c ] ÷ (75), 254 [ M - O A c - HOAc] ÷ (60), 253, (23), 242 (25), 226 (22), 225 (24), 224 (27), 202 (41), 201 (37). ~H NMR (500 MHz, CDCI3): Table 3; Z3C NMR (50 MHz, CDCI3): Table 1. 1-Epideacetylbowdensine (7). Found: C, 62.78; H, 6.65; N, 4.45. CtTH21NO 5 requires: C, 63.95; H, 6.58; N, 4.39%. Mp 162-164 °. [a] 22 + 22 ° (CHC13; c 0.47). CD [0]256 + 4520, [0]282 - 7 5 . IR /')max cm-I: 34503200 (-OH), 2924, 1618, 1473, 1373, 1274, 1215, 1045, 939, (-OCH20-), 755. EIMS 70 eV, m/z (rel. int.): 319 [M] ÷ (100), 302 (12), 275 (29), 246 (20), 232 (76), 220 (25), 219 (22), 203 (18), 57 (38), 56 (25). 'H NMR (500 MHz, CDCI3): Table 3; ~3C NMR (50 MHz, CDC13): Table 1. 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