You are on page 1of 2

Journal of Ethnopharmacology 99 (2005) 145146

Ethnopharmacological communication
Two avonoids from Artemisia herba-alba Asso with in vitro
GABA
A
-benzodiazepine receptor activity
Sam Medhat Salah, Anna Katharina J ager

Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, 2 Universitetsparken, 2100 Copenhagen O, Denmark
Received 9 December 2004; received in revised form 17 January 2005; accepted 17 January 2005
Abstract
An ethyl acetate extract of Artemisia herba-alba was partitioned by HPLCin 10 fractions that were tested in the [
3
H]-umazenil radioligand
assay, for afnity to the GABA
A
-benzodiazepine receptor. Two fractions showed activity from which hispidulin and cirsilineol were isolated.
The structures were conrmed by
1
H NMR. The IC
50
values were 8 M for hispidulin and 100 M for cirsilineol.
2005 Elsevier Ireland Ltd. All rights reserved.
Keywords: Artemisia herba-alba; Cirsilineol; GABA
A
-benzodiazepine receptor; Hispidulin
1. Plant
Aerial parts of Artemisia herba-alba Asso (Lamiaceae)
were collected fromLebanese herbal stores (dabbous stores),
in Beirut, Lebanon. The plant was identied with help from
K. Sleem, American University in Beirut. A voucher sample
(Salah1) is stored at the Department for Medicinal Chemistry,
The Danish University of Pharmaceutical Sciences.
2. Uses in traditional medicine
Artemisia herba-alba has a wide use in traditional
medicine, for treatment of gastric disturbances, such as diar-
rhoea, abdominal cramps and for healing external wounds
(Feuerstein et al., 1986). Uses for diabetes mellitus and
other conditions as jaundice are also reported (Marrif et al.,
1995). The species is recommended for neurological dis-
orders, and an ethanolic extract has shown activity in the
GABA
A
-benzodiazepine receptor assay (Salah and J ager,
2005).

Corresponding author.
E-mail address: ankj@dfuni.dk (A.K. J ager).
3. Previously isolated classes of constituents
Flavonoids, essential oils and eudesmanolides (Feuerstein
et al., 1986; Saleh et al., 1987; Boriky et al., 1996).
4. Material and methods
Ten gram of Artemisia herba-alba plant material was rst
extracted with heptane to remove non-active lipophilic com-
pounds. The plant material was hereafter extracted with 3
100 ml ethyl acetate, which subsequently was partitioned
against 3 100 ml water to remove non-active polar com-
pounds.
The separation of the ethyl acetate phase was car-
ried out by gradient HPLC on a Phenomenex Luna 5
C-18 column (250 mm21.2 mm), using MeCN/0.05%
TFA as mobile phase. The resulting 10 fractions were
tested in the GABA
A
-benzodiazepine receptor assay, where
the afnity to the benzodiazepine site was measured
by the [
3
H]-umazenil radioligand assay according to
Kahnberg et al. (2002) and Risa et al. (2004).
1
H NMR
was used to assign the structures of the two active
compounds.
0378-8741/$ see front matter 2005 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2005.01.031
146 S.M. Salah, A.K. J ager / Journal of Ethnopharmacology 99 (2005) 145146
5. Results
Two fractions were active in the GABA
A
-benzodiazepine
receptor assay. From these fractions hispidulin (1.2 mg) and
cirsilineol (1.0 mg) were isolated and identied on basis
of
1
H NMR by comparison with data reported previously
(Matsuura et al., 1973; Akkal et al., 2003).
The IC
50
values for hispidulin and cirsilineol were 8 and
104 M, respectively. The IC
50
value of hispidulin in the
present studyis withinthe same range as previouslypublished
values of 1.3 M (Shen et al., 1994; Kavvadias et al., 2003).
Acknowledgements
The authors thank the Danish Medical Research Council
for nancial support and K. Sleem, American University in
Beirut, for identication of the plant material.
References
Akkal, S., Benayache, F., Bentamene, A., Medjroubi, K., Seguin, E.,
Tillequin, F., 2003. Flavonoid aglycones from Centaurea napifolia.
Chemistry of Natural Compounds 39, 219220.
Boriky, D., Berrada, M., Talbi, M., Keravis, G., Rouessac, F., 1996.
Eudesmanolides from Artemisia herba-alba. Phytochemistry 43,
309311.
Feuerstein, I., M uller, D., Hobert, K., Danin, A., Segal, R., 1986. The
constituents of essential oils from Artemisia herba-alba population of
Israel and Sinai. Phytochemistry 25, 23432347.
Kahnberg, P., Lager, E., Rosenberg, C., Schougaard, J., Camet, L., Sterner,
O., Nielsen, E.., Nielsen, M., Liljefors, T., 2002. Renement and
evaluation of a pharmacophore model for avone derivatives bind-
ing to the benzodiazepine bite of the GABA
A
receptor. Journal of
Medicinal Chemistry 45, 41884201.
Kavvadias, D., Monshein, V., Sand, P., Riederer, P., Schreier, P.,
2003. Constituents of Sage (Salvia ofcinalis) with in vitro afn-
ity to human brain benzodiazepine receptor. Planta Medica 69, 113
117.
Marrif, H.I., Ali, B.H., Hassan, K.M., 1995. Some pharmacological stud-
ies on Artemisia herba-alba (Asso) in rabbits and mice. Journal of
Ethnopharmacology 49, 5155.
Matsuura, S., Kunii, T., Matsuura, A., 1973. Synthetic studies of the
avone derivatives. Synthesis of cirsiliol and cirsilineol. Chemical
and Pharmaceutical Bulletin 21, 27572759.
Risa, J., Risa, A., Adsersen, A., Gauguin, B., Stafford, G.I., van Staden,
J., J ager, A.K., 2004. Screening of plants used in southern Africa
for epilepsy and convulsions in the GABA
A
-benzodiazepine receptor
assay. Journal of Ethnopharmacology 93, 177182.
Salah, S.M., J ager, A.K., 2005. Screening of traditionally used Lebanese
herbs for neurological activities. Journal of Ethnopharmacology 97,
145149.
Saleh, N.A.M., El-Negoumy, S.I., Abou-Zaid, M.M., 1987. Flavonoids of
Artemisia judaica, A. monosperma and Artemisia herba-alba. Phyto-
chemistry 26, 30593064.
Shen, X.L., Nielsen, M., Witt, M.R., Sterner, O., Bergendorff, O., 1994.
Inhibition of [methyl-
3
H]diazepam binding to rat brain membranes
in vitro by diantin and skrofulein. Acta Pharmacologica Sinica 15,
385388.