Fitoterapia 71 Ž2000.

101]104

A new steroidal alkaloid from the seeds of

Holarrhena antidysenterica
A. Kumar, M. AliU
Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Jamia Hamdard
(Hamdard Uni¨ ersity), Hamdard Nagar, New Delhi 110 062, India

Received 29 June 1999; accepted in revised form 10 August 1999

Abstract

A new steroidal alkaloid, named antidysentericine, has been isolated from the seeds of
Holarrhena antidysenterica and characterized as 3b-dimethylaminocon-5-enin-18-one Ž1..
Q 2000 Elsevier Science B.V. All rights reserved.

Keywords: Holarrhena antidysenterica; Alkaloids

1. Introduction

Holarrhena antidysenterica L., syn. H. pubescens ŽBuch.-Ham.. Wall.

ŽApocynaceae., is a typical Indian medicinal plant. The bark and seeds are used to
treat amoebic dysentery, diarrhoea, asthma, bronchopneumonia, malaria and some
other disorders w1x. Two bibliographies on the work done on different aspects of
this plant have been published and the isolation of a number of steroidal con-
stituents has been reported w2,3x. On account of its interesting chemistry and

U
Corresponding author.
E-mail address: m.ali@mail.usa.com ŽM. Ali.

0367-326Xr00r$ - see front matter Q 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 3 6 7 - 3 2 6 X Ž 9 9 . 0 0 1 1 1 - 2
102 A. Kumar, M. Ali r Fitoterapia 71 (2000) 101]104

biological activity, H. antidysenterica is still the subject of a great deal of interest

w4,5x. This communication describes the isolation and structure elucidation of a
new steroidal alkaloid, antidysentericine Ž1., from the seeds of the plant.

2. Results and discussion

Compound 1, named antidysentericine, was obtained by Si-gel column chro-

matography as colourless crystals. Its IR spectrum showed characteristic bands for
secondary amino Ž3390 cmy1 . and five membered cyclic amide Ž1740 cmy1 . groups.
Its mass spectrum showed a molecular ion peak at m r z 356 consistent with the
molecular formula C 23 H 36 N2 O. The appearance of the base peak at m r z 71
wC 3 H 5 NOxq and the fragments at m r z 137 wC 8,14 ]C 12,13 fissionx, 122 w137-Mexq
and 66 w137]71xq suggested the presence of a 18-keto-18,20-epimino group charac-
teristic of alkaloids of the conanine series w6x. The spectrum also showed ion peaks
at m r z 110 wC 2,3 ]C 5,10 ]C 7,8 fissionxq, 96 w110-CH 2 xq, 99, 257 wC 1,10 ]C 4,5 fissionxq,
55 w99-NMe 2 xq, 84 w99-Mexq, 69 w84-Mexq, 120 w257-137xq, 186 w257-71xq, 219
wM-137xq, 341 wM-Mexq, 312 wM-NMe 2 xq, suggesting the existence of NMe 2 group
in ring A at C-3 and tri-substituted olefinic linkage at C-5. The 1 H-NMR spectrum
of 1 displayed a one-proton double doublet at d 5.45 Ž J 4.79, 4.81 Hz. for C-6
vinylic proton, a C-3 methine multiplet at d 3.16 and a one-proton double quartet
at d 3.50 assigned to C-20 axial methine proton. A three-proton deshielded doublet
at d 1.42 Ž J 6.54 Hz. was ascribed to C-21 methyl function. A three-proton singlet
at d 0.94 was accounted to C-19 tertiary methyl group. Two three-proton deshielded
signals at d 2.68 and 2.66 confirmed the attachment of methyl groups with the
secondary amino function at C-3. The chemical shifts were compared with those of
conessine derivatives w7x. A D 2 O exchangeable NH proton was discernible at d
2.94. The remaining methine and methylene signals resonated between d 2.45 and
1.09. Acetylation of 1 yielded a mono acetyl derivative. Treatment of 1 with a
mixture of formaldehyde and formic acid produced 18-oxoconessine. Based on
these observations, the structure of antidysentericine has been elucidated as
3b-dimethylaminocon-5-enin-18-one Ž1..
 A. Kumar, M. Ali r Fitoterapia 71 (2000) 101]104 103

3. Experimental

3.1. Plant material

The seeds of H. antidysenterica were procured from a local market and authenti-
cated by Dr M.P. Sharma, Reader, Botany Department of this university. A
voucher specimen was deposited in the Phytochemistry Research Laboratory,
Faculty of Pharmacy, Jamia Hamdard, New Delhi.

3.2. Extraction and isolation

The seeds were dried in an oven at a temperature below 508C and coarsely
powdered. The ground seeds Ž4.5 kg. were Soxhlet extracted with petrol Ž60]808C.
and concentrated to yield 750 ml of reddish-brown oil. The marc was re-extracted
with 95% EtOH to yield 280 g of dark brown viscous mass which was treated with
dilute HCl. The soluble portion was washed with CHCl 3 , made alkaline ŽpH 9. with
ammonium hydroxide and extracted with CHCl 3 . After evaporation of the solvent
to dryness, the viscous dark brown mass Žtotal alkaloid residue. was chro-
matographed over Si-gel column. Elution with CHCl 3-MeOH 95:5 and recrystal-
lization from acetone gave 37 mg Žyield: 0.00082%. of 1 Žantidysentericine ., mp
118]1198; w a x 22
D
:-24.5 ŽCHCl 3 , c 0.1.; UV max ŽMeOH.: 222 nm Žlog g 6.5.; IR
bands ŽKBr.: 3390, 2940, 2855, 1740, 1465, 1375, 1255, 1080, 1055, 725 cmy1 ;
1
H-NMR Ž300 MHz, CDCl 3 .: d 5.45 Ž1H, dd, J 4.79, 4.81 Hz, H-6a ., 3.50 Ž1H, d, J
12.06 Hz, H-20a ., 2.94 Ž1H, s, D 2 O exchangeable, NH., 2.76 Ž1H, m, H-9a ., 2.68
Ž3H, s, NMe 2 ., 2.45 Ž1H, m, H-7a ., 2.42 Ž1H, m, H-7b ., 2.18 Ž1H, d, J 9.09 Hz,
H-4b ., 2.10 Ž1H, J 3.81 Hz, H-4a ., 2.07 Ž1H, dddd, J 5.28, 2.83, 2.39, 10.5 Hz,
H-12a ., 1.93 Ž1H, m, H-14a ., 1.77 Ž1H, m, H-8b ., 1.67 Ž1H, m, H-11a ., 1.63 Ž1H,
m, H-2a ., 1.52 Ž1H, m, H-15a., 1.47 Ž1H, m, H-16a., 1.42 Ž1H, d, J 6.54, H-21a .,
1.36 Ž1H, dddd, J 6.54, 12.41, 5.61, 4.87 Hz, H-1a ., 1.31Ž1H, m, H-11b ., 1.25 Ž1H,
m, H-15a., 1.22 Ž1H, dddd, J 4.84, 11.12, 9.96, 3.27 Hz, H-1b ., 1.14 Ž1H, m, H-2b .,
1.13 Ž1H, m, H-17a ., 1.09 Ž1H, m, H-16a., 0.94 Ž3H, s, Me-19.; EIMS m r z Žrel.
int..: 356 ŽMq 56.1%. ŽC 23 H 36 ON2 ., 341 Ž22.5., 312 Ž3.0., 257 Ž3.4., 219 Ž4.0., 186
Ž2.0., 137 Ž5.8., 122 Ž6.3., 120 Ž13.5., 110 Ž5.5., 99 Ž3.6., 96 Ž11.3., 84 Ž99.9., 71 Ž100.,
69 Ž56.7., 66 Ž10.6., 56 Ž24.4., 55 Ž21.1..

Acknowledgements

Thanks are due to the Head, Instrumentation Centre, Indian Institute of

Technology, New Delhi for recording NMR spectra.

References
w1x Arya Vaidya Sala, Kottakkal. Indian medicinal plants, vol. 3. Hyderabad, AP, India: Orient
Longman, 1995:156.
104 A. Kumar, M. Ali r Fitoterapia 71 (2000) 101]104

w2x MAPA, Medicinal and Aromatic Plants Abstract. Bibliography on Holarrhena antidysenterica
ŽLinn.. Wall ŽApocynaceae.. New Delhi, India: NISCOM, 1982;4:351.
w3x Chaturvedi GN, Singh KP, Gupta JP. Indian Med Gaz 1981;179.
w4x Siddiqui BS, Usmani SB, Begum S, Siddiqui S. Nat Prod Rep 1995;12:361.
w5x Ali M, Gupta J. Apmak Pharm 1994;7:140.
w6x Budzikiewicz H, Djerassi C, Williams DH. Interpretation of mass spectra of organic compounds.
San Francisco: Holden-Day, 1964:33.
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