390 PlantaMed. 57 (1991)
D. Uhrin', B. Proksa"3, andJ. Zhamiansan2 1
2
Institute of Chemistry, Slovak Academy of Sciences,
842 38 Bratislava, Czechoslovakia Institute of Chemistry, Mongolian Academy of Sciences, than Bator, Mongolia Address for correspondence
Received: July 8, 1990
The diterpenoid alkaloids aconitine, deoxyaconitine, hypaconitine, mesaconitine, and beiwutine have
been isolated from the roots of Aconitwn kusnezoffli Reichb. (Ranunculaceae) (1). Besides these bases, we have isolated compounds 1 and 2 from the aerial parts of A. kusnezoffli collected in Mongolia.
The MS of alkaloid I showed M at ,n/z 359 (C22H33N03), which, after loss of H20, afforded the base peak at ,n/z 341. Acetylation of 1 with acetic anhydridepyridine yielded the triacetate 3 (M at m./z 485). A simpler
MS fragmentation pattern was encountered with compound 2, where the molecular radical ion at rn/z 343 (C22H33N03) was also the base peak. TheIR spectra of 1 and
2 disclosed bands characteristic of 0-H, C-H and C=C. The analyses of 1H- and 13C-NMR spectra and comparison of the
shift data published for 1i-acetyl-i,19-epoxydenudatine
was placed in an equatorial position at C-i. The 1H-NMR
signals (Table 2) of alkaloid I were assigned by a 2Dheterocorrelated experiment; the H-H coupling constants and the chemical shift data were obtained by a series of both homodecoupling and 1D COSY experiments (5). The relative configuration at C-li and C-15 and assignment of methylene protons were achieved by a NOE differential experiment (6). The difference in chemical shifts of H-il in the spectra of 1 and 2 (ó = 4.28 vs. 3.59 ppm) is due to an interaction of this proton with oxygen of the C-i-OH group in Table 1 '3C-NMR chemical shifts ofcompounds 1 and 2 (DMSO-d6). C
1
2 26.1 20.3 39.8 33.5 51.7 22.5 46.6
1
68.7
2 3
30.6
4
33.0
5 6 7
51.9 22.8
8
43.0 52.8
43.1
9 10
50.3
45.0
11 12 13 14 15 16 17 18 19 20
71.7 41.4 24.1 27.1 76.3 153.9 107.9 26.0 56.8 66.8
24.0 27.7 76.7 154.2 108.5 26.5
50.1
50.1
13.3
13.5
38.5
46.4
NCH2CH3 NCH2CH3
52.2 71.6
41.8
57.1
71.0
(2) indicated that both compounds possess the atisine skeleton and differ in the number of hydroxy groups. Table 2
'H.NMR chemical shifts of compounds 1 and 2 (DMSO-d5).
2
1
Proton
1
2a 2b 3a 3b 5 6a 6b 7 9 1 R1
OH, R2 H
2 R1 R2 H 3 R' H3C—CO—O—, R2 H3C—CO—
In the 13C-NMR of I and 2 (Table 1) significant differences in chemical shifts were observed in signals associated with the A-ring carbons. These signals were assigned by a semi-selective INEPT experiment (3). Magnetization transfer through H-18 protons revealed signals of C3, C-4, C-5, and C-i 9; magnetization transfer from H-20 re-
sulted in resonances of carbons C-5, C-b, and C-i9. According to these data, as well as the published data of substituent effects of a hydroxy group on 13C chemical shifts in
cyclohexane derivatives (4), the third hydroxy group in 1
11
12 13a 13h 14a 14b 15
l7a I 7b
18 19a 19b 20 NCH2CH3 NCFI2CH3 C-11--OT!
C-iS--OH
J(Hz)
3.98(m) 2.30(m) 1.56(m) 1.19(m) 1.49(m) 1.15(d) 2.80(dd) 1.08(dd) 1.99(m) 1.25(d) 4.28 (dd) 1.99(m) 1.31(m) 1.55(m) 0.97(m) 1.84(m)
3.61 (s) 2.40 (ABq, d) 0.99 (t)
4.02(d) 4.87(d)
Not identified.
J(Hz)
1.84(2H,m)
7.50 13.7, 7.8
13.7,5.7 9.1 9.0,4.4
4.07 (m)
5.06(dd) 4.78 (dd) 0.65(s) 2.18(d) 2.42(d)
ô
2.3,2.3 2.3,2.3 10.7 10.7
12.1,7.3 7.3 4.7 6.0
1.12(m) 1.50(m) 1.06(d) 2.74(dd) 1.08(dd) 1.95(d) 1.25(d) 3.59 (dd)
2.00(m) 1.31(m) 1.57(m) 0.97(m) 1.80(m) 4.05 (ddd) 5.07(dd) 4.79 (dd)
0.67(s) 2.16(dd) 2.41(d) 3.26(s) 2.37 (ABQ, d)
0.97 (t)
4.19(d) 4.88(d)
8.3 13.8,8.3 13.8,5.3 5.27 9.6 9.6,4.4
5.9,2.4, 1.9
2.7,1.9 2.7,2.4 10.8,1.9 10.8
12.1,7.3 7.3 4.4 5.9
Downloaded by: University of Illinois. Copyrighted material.
Lepenine and Denudatine: New Alkaloids from A conitum kusnezoffli
Letters
Planta Med. 57(1991) 391
Letters
the structure of 1. A similar deshielding was observed e.g. in the spectra of non-reducing dianhydrides of a-n-galacturonic acid (7). In accord with these data alkaloid I is iden-
tical with lepenine, previously isolated from A. pseudohuiliense (8) and base 2 has the same structure as denudatine (9). In preliminary pharmacological tests denudatine (2) decreased the tone and peristalsis of the dog intestine (10); lepenine (1) showed antihypoxic activity and enhanced the action of clinically used hypnotics.
Materials and Methods
Volatile Constituents of the Rhizome of Homalomena occulta Cheng-MingZhou"3, Chuan Yao1, Hai-Lin Sun2, Sheng-Xiang Qiu1, and Guo-Yin Cui1 1
National Institutes of Pharmaceutical Research and Development, Shahe, Beijing 102206, People's Republic of China 2 Beijing Medicinal Material Corporation, Beijing, People's Republic of China
Address for correspondence Received: December 6, 1990
The respective spectra were recorded with following instruments: El-MS. Jeol JMS 100D (70 eV, 300 1i.A); 1H- and
The plant material was collected in the region Erdene-Tsagaan in Mongolia in September 1988; a voucher is depo-
sited in the Herbarium of the Institute of Chemistry, Mongolian Academy of Sciences, Ulan Bator.
Isolation of alkaloids The air-dried aerial parts ofA. kusnezoffli (250 g) were extracted with methanol (3 x 11), the combined extracts were evaporated, the residue was dissolved in 0.5 mol/l HC1 (100 ml). the solution was filtered, the pH of the filtrate was adjusted to 8.5 and the precipitated mixture was collected and dried to yield 0.9 g of alkaloids. This mixture was chromatographed on alumina by elution
with chloroform containing an increasing amount of methanol. Fractions were evaluated by TLC (silica gel, CHC13 — CH3OH — CH3COCH3 — NH4OH 42 : 50: 8 : 5). The fraction characterized by RF
The dry rhizome of Homalomena occulta (Lour.) Schott (Araceae) is a famous traditional Chinese
medicinal plant used for several hundred years. The rhizome of this plant contains 0.79% essential oil, the oil is yellow and has a characteristic odour, the density of D11° = = 1.4692. In the present work, the essential oil 0.9880,
of Homalomena occulta rhizome was studied by gas
chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Fourty-four compounds were identified
from the oil. Table I shows the detailed results of the analyses of the oil. The major compounds are linalool, terpi-
nen-4-oi, cedrenol, saussurea lactone, ô-cadinol, a-ter-
pineol, eremophilene, moslene. These are similar to the resuits reported in previous publication (1, 2, 4). The rhizome of Hornalomena occulta evidently is an abundant and valuable source of linalol and terpinen-4-oi.
0.20 on TLC sheets was evaporated and the residue was crystallized from methanol — acetone (1 : 1) to give alkaloid 1 (45 mg), m.p.
199—201 °C, IR (KBr) 3380. 3094, 2919, 2880, 1650, 1485 cm1. MS m.z (%): 359 (60), 358 (37), 341 (100), 330 (23), 324(11), 313 (20), 300 (30), 247 (15), 230 (18). Fraction of RF 0.25 crystallized from hexane — acetone (1: 1), to furnish base 2(28mg), m.p. 249—
Table 1
251°C; IR (KBr): 3378, 3091, 2992, 2965, 1649, 1458cm'. MS ,n/z (%): 343 (100), 342 (13), 325 (18), 300 (10), 231 (8), 210 (6).
fi-pinene
The 1H- and 13C-NMR data of 1 and 2 are listed in Tables 1 and 2, respectively.
References
Composition of the essential oil of Homalornena occulta rhizome.
Compounds
a-phellandrene
1.57 1.69
moslene a-terpinene
2.22 2.20
f3-phellandrene A3-carene
1.67
la-terpineol
0.81
linalool
Wang, Y. G., Zhu, Y. L., Zhu, R. H. (1980) Yao Hsueh Hsueh Pao 15, 526. 2 Proksa, B., UhrIn, D., Batsuren, D., Batbaiar, N., Selenge, D. (1990) Planta Med. 56, 461. Bax, A. (1984) J. Magn. Reson. 57, 314. Schneider, H. J., Hoppen, V. (1978) J. Org. Chem. 43, 3866. Kessler, H., Oschkinat. H.. Griesinger, C. (1986) J. Magn. Reson. 70, 106. 6 Kövér, K. E. (1984)J. Magn. Reson. 59, 485. Capek, P., UhrIn, D., Rosik, J., Kardoiovk, A., Toman, R., Mihálov, V. (1988) Carbohyd. Res. 182, 160. 8 Han, G. Q., Chen, Y. Y., Liang, X. T. (1988) in: The Alkaloids, (Brossi, A., ed.), Academic Press, New York, Vol. 32, p. 245. Pelletier, S. W.. Mody, N. V. (1981) in: The Alkaloids, (Manske, H. H. F., Rodrigo, G. A., eds.), Academic Press, New York, Vol. 18, p. 131. 10 Singh, N., Chopra, K. L. (1962) J. Pharm. Pharmacol. 14, 286.
Percentages
(2-methoxyethoxy)-benzene terpinen-4-ol a-terpineol
1.31
36.88 0.65
0.22 2.88
eucarvone
0.99
rhodinal geraniol methyl ether piperitone
1.31 1.81
anethole
0.46 0.42
methyl nerate geraniol
a-humulene
a-cedrene y-cadinene a-muurolene y-muurolene
a-copaene eremophilene driminol spiro[4,4inonane-2-one ô-cadinol
cedrenol saussurea lactone 1,2,9,10-tetrahydroaristolane
0.46 1.02 0.47
0.40 0.44 0.67 0.73 1.86
2.46 0.53 0.59 3.21 4.34 4.67 0.68
Downloaded by: University of Illinois. Copyrighted material.
13C-NMR, Bruker AM 300 operating at 300 and 75MHz, respectively; IR, Perkin Elmer, model 983.
D. Uhrin', B. Proksa"3, andJ. Zhamiansan2 1
2
Institute of Chemistry, Slovak Academy of Sciences,
842 38 Bratislava, Czechoslovakia Institute of Chemistry, Mongolian Academy of Sciences, than Bator, Mongolia Address for correspondence
Received: July 8, 1990
The diterpenoid alkaloids aconitine, deoxyaconitine, hypaconitine, mesaconitine, and beiwutine have
been isolated from the roots of Aconitwn kusnezoffli Reichb. (Ranunculaceae) (1). Besides these bases, we have isolated compounds 1 and 2 from the aerial parts of A. kusnezoffli collected in Mongolia.
The MS of alkaloid I showed M at ,n/z 359 (C22H33N03), which, after loss of H20, afforded the base peak at ,n/z 341. Acetylation of 1 with acetic anhydridepyridine yielded the triacetate 3 (M at m./z 485). A simpler
MS fragmentation pattern was encountered with compound 2, where the molecular radical ion at rn/z 343 (C22H33N03) was also the base peak. TheIR spectra of 1 and
2 disclosed bands characteristic of 0-H, C-H and C=C. The analyses of 1H- and 13C-NMR spectra and comparison of the
shift data published for 1i-acetyl-i,19-epoxydenudatine
was placed in an equatorial position at C-i. The 1H-NMR
signals (Table 2) of alkaloid I were assigned by a 2Dheterocorrelated experiment; the H-H coupling constants and the chemical shift data were obtained by a series of both homodecoupling and 1D COSY experiments (5). The relative configuration at C-li and C-15 and assignment of methylene protons were achieved by a NOE differential experiment (6). The difference in chemical shifts of H-il in the spectra of 1 and 2 (ó = 4.28 vs. 3.59 ppm) is due to an interaction of this proton with oxygen of the C-i-OH group in Table 1 '3C-NMR chemical shifts ofcompounds 1 and 2 (DMSO-d6). C
1
2 26.1 20.3 39.8 33.5 51.7 22.5 46.6
1
68.7
2 3
30.6
4
33.0
5 6 7
51.9 22.8
8
43.0 52.8
43.1
9 10
50.3
45.0
11 12 13 14 15 16 17 18 19 20
71.7 41.4 24.1 27.1 76.3 153.9 107.9 26.0 56.8 66.8
24.0 27.7 76.7 154.2 108.5 26.5
50.1
50.1
13.3
13.5
38.5
46.4
NCH2CH3 NCH2CH3
52.2 71.6
41.8
57.1
71.0
(2) indicated that both compounds possess the atisine skeleton and differ in the number of hydroxy groups. Table 2
'H.NMR chemical shifts of compounds 1 and 2 (DMSO-d5).
2
1
Proton
1
2a 2b 3a 3b 5 6a 6b 7 9 1 R1
OH, R2 H
2 R1 R2 H 3 R' H3C—CO—O—, R2 H3C—CO—
In the 13C-NMR of I and 2 (Table 1) significant differences in chemical shifts were observed in signals associated with the A-ring carbons. These signals were assigned by a semi-selective INEPT experiment (3). Magnetization transfer through H-18 protons revealed signals of C3, C-4, C-5, and C-i 9; magnetization transfer from H-20 re-
sulted in resonances of carbons C-5, C-b, and C-i9. According to these data, as well as the published data of substituent effects of a hydroxy group on 13C chemical shifts in
cyclohexane derivatives (4), the third hydroxy group in 1
11
12 13a 13h 14a 14b 15
l7a I 7b
18 19a 19b 20 NCH2CH3 NCFI2CH3 C-11--OT!
C-iS--OH
J(Hz)
3.98(m) 2.30(m) 1.56(m) 1.19(m) 1.49(m) 1.15(d) 2.80(dd) 1.08(dd) 1.99(m) 1.25(d) 4.28 (dd) 1.99(m) 1.31(m) 1.55(m) 0.97(m) 1.84(m)
3.61 (s) 2.40 (ABq, d) 0.99 (t)
4.02(d) 4.87(d)
Not identified.
J(Hz)
1.84(2H,m)
7.50 13.7, 7.8
13.7,5.7 9.1 9.0,4.4
4.07 (m)
5.06(dd) 4.78 (dd) 0.65(s) 2.18(d) 2.42(d)
ô
2.3,2.3 2.3,2.3 10.7 10.7
12.1,7.3 7.3 4.7 6.0
1.12(m) 1.50(m) 1.06(d) 2.74(dd) 1.08(dd) 1.95(d) 1.25(d) 3.59 (dd)
2.00(m) 1.31(m) 1.57(m) 0.97(m) 1.80(m) 4.05 (ddd) 5.07(dd) 4.79 (dd)
0.67(s) 2.16(dd) 2.41(d) 3.26(s) 2.37 (ABQ, d)
0.97 (t)
4.19(d) 4.88(d)
8.3 13.8,8.3 13.8,5.3 5.27 9.6 9.6,4.4
5.9,2.4, 1.9
2.7,1.9 2.7,2.4 10.8,1.9 10.8
12.1,7.3 7.3 4.4 5.9
Downloaded by: University of Illinois. Copyrighted material.
Lepenine and Denudatine: New Alkaloids from A conitum kusnezoffli
Letters
Planta Med. 57(1991) 391
Letters
the structure of 1. A similar deshielding was observed e.g. in the spectra of non-reducing dianhydrides of a-n-galacturonic acid (7). In accord with these data alkaloid I is iden-
tical with lepenine, previously isolated from A. pseudohuiliense (8) and base 2 has the same structure as denudatine (9). In preliminary pharmacological tests denudatine (2) decreased the tone and peristalsis of the dog intestine (10); lepenine (1) showed antihypoxic activity and enhanced the action of clinically used hypnotics.
Materials and Methods
Volatile Constituents of the Rhizome of Homalomena occulta Cheng-MingZhou"3, Chuan Yao1, Hai-Lin Sun2, Sheng-Xiang Qiu1, and Guo-Yin Cui1 1
National Institutes of Pharmaceutical Research and Development, Shahe, Beijing 102206, People's Republic of China 2 Beijing Medicinal Material Corporation, Beijing, People's Republic of China
Address for correspondence Received: December 6, 1990
The respective spectra were recorded with following instruments: El-MS. Jeol JMS 100D (70 eV, 300 1i.A); 1H- and
The plant material was collected in the region Erdene-Tsagaan in Mongolia in September 1988; a voucher is depo-
sited in the Herbarium of the Institute of Chemistry, Mongolian Academy of Sciences, Ulan Bator.
Isolation of alkaloids The air-dried aerial parts ofA. kusnezoffli (250 g) were extracted with methanol (3 x 11), the combined extracts were evaporated, the residue was dissolved in 0.5 mol/l HC1 (100 ml). the solution was filtered, the pH of the filtrate was adjusted to 8.5 and the precipitated mixture was collected and dried to yield 0.9 g of alkaloids. This mixture was chromatographed on alumina by elution
with chloroform containing an increasing amount of methanol. Fractions were evaluated by TLC (silica gel, CHC13 — CH3OH — CH3COCH3 — NH4OH 42 : 50: 8 : 5). The fraction characterized by RF
The dry rhizome of Homalomena occulta (Lour.) Schott (Araceae) is a famous traditional Chinese
medicinal plant used for several hundred years. The rhizome of this plant contains 0.79% essential oil, the oil is yellow and has a characteristic odour, the density of D11° = = 1.4692. In the present work, the essential oil 0.9880,
of Homalomena occulta rhizome was studied by gas
chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Fourty-four compounds were identified
from the oil. Table I shows the detailed results of the analyses of the oil. The major compounds are linalool, terpi-
nen-4-oi, cedrenol, saussurea lactone, ô-cadinol, a-ter-
pineol, eremophilene, moslene. These are similar to the resuits reported in previous publication (1, 2, 4). The rhizome of Hornalomena occulta evidently is an abundant and valuable source of linalol and terpinen-4-oi.
0.20 on TLC sheets was evaporated and the residue was crystallized from methanol — acetone (1 : 1) to give alkaloid 1 (45 mg), m.p.
199—201 °C, IR (KBr) 3380. 3094, 2919, 2880, 1650, 1485 cm1. MS m.z (%): 359 (60), 358 (37), 341 (100), 330 (23), 324(11), 313 (20), 300 (30), 247 (15), 230 (18). Fraction of RF 0.25 crystallized from hexane — acetone (1: 1), to furnish base 2(28mg), m.p. 249—
Table 1
251°C; IR (KBr): 3378, 3091, 2992, 2965, 1649, 1458cm'. MS ,n/z (%): 343 (100), 342 (13), 325 (18), 300 (10), 231 (8), 210 (6).
fi-pinene
The 1H- and 13C-NMR data of 1 and 2 are listed in Tables 1 and 2, respectively.
References
Composition of the essential oil of Homalornena occulta rhizome.
Compounds
a-phellandrene
1.57 1.69
moslene a-terpinene
2.22 2.20
f3-phellandrene A3-carene
1.67
la-terpineol
0.81
linalool
Wang, Y. G., Zhu, Y. L., Zhu, R. H. (1980) Yao Hsueh Hsueh Pao 15, 526. 2 Proksa, B., UhrIn, D., Batsuren, D., Batbaiar, N., Selenge, D. (1990) Planta Med. 56, 461. Bax, A. (1984) J. Magn. Reson. 57, 314. Schneider, H. J., Hoppen, V. (1978) J. Org. Chem. 43, 3866. Kessler, H., Oschkinat. H.. Griesinger, C. (1986) J. Magn. Reson. 70, 106. 6 Kövér, K. E. (1984)J. Magn. Reson. 59, 485. Capek, P., UhrIn, D., Rosik, J., Kardoiovk, A., Toman, R., Mihálov, V. (1988) Carbohyd. Res. 182, 160. 8 Han, G. Q., Chen, Y. Y., Liang, X. T. (1988) in: The Alkaloids, (Brossi, A., ed.), Academic Press, New York, Vol. 32, p. 245. Pelletier, S. W.. Mody, N. V. (1981) in: The Alkaloids, (Manske, H. H. F., Rodrigo, G. A., eds.), Academic Press, New York, Vol. 18, p. 131. 10 Singh, N., Chopra, K. L. (1962) J. Pharm. Pharmacol. 14, 286.
Percentages
(2-methoxyethoxy)-benzene terpinen-4-ol a-terpineol
1.31
36.88 0.65
0.22 2.88
eucarvone
0.99
rhodinal geraniol methyl ether piperitone
1.31 1.81
anethole
0.46 0.42
methyl nerate geraniol
a-humulene
a-cedrene y-cadinene a-muurolene y-muurolene
a-copaene eremophilene driminol spiro[4,4inonane-2-one ô-cadinol
cedrenol saussurea lactone 1,2,9,10-tetrahydroaristolane
0.46 1.02 0.47
0.40 0.44 0.67 0.73 1.86
2.46 0.53 0.59 3.21 4.34 4.67 0.68
Downloaded by: University of Illinois. Copyrighted material.
13C-NMR, Bruker AM 300 operating at 300 and 75MHz, respectively; IR, Perkin Elmer, model 983.
