Sociedade Brasileira de Química (SBQ)
Triterpene barbinervic
(Mirtaceae)
acid
isolated
1
from
Eugenia
2
punicifolia
2
3
Kátia Gomes de Lima Araújo (PQ), Benjamin Rodriguez (PQ), Begoña Bartolomé (PQ), Luis Gandía (PQ) and
1*
1
Wilson C. Santos (PQ) Programa de Pós Graduação em Ciências Aplicadas a Produtos Para Saúde, Faculdade
2
de Farmácia, Universidade Federal Fluminense, Niterói, RJ; Consejo Superior de Investigaciones Científicas,
3
Madrid; Departamento de Farmacología, Universidad Autónoma de Madrid, España
*
[email protected]
Key words: Eugenia punicifolia, barbinervic acid, cholinergic neurotransmission
Introduction
Myrtaceae family is constituted by more than 3.000
species and largely distributed in the Brazilian
tropical Amazon region with the genus Syzigium and
Eugenia commonly used for diarrhea and stomach
1,2
disturbance, and as hypoglycemic medicament .
The
mechanism
underlying
pharmacological
properties of the genus Eugenia may be partly
related to flavonoids (myricitrin, quercetin, and
quercetrin), steroids, terpenoids, tanines, and
3
anthraquinones , but Eugenia punicifolia aqueous
extract was able to augment cholinergic nicotinic
neurotransmission in rat diaphragm muscle endplate
4
model . Furthermore, we have shown that E.
punicifolia dichloromethane extract enhanced the
cholinergic neurotransmission in adrenal chromaffin
5
cells , and also decreased production of
inflammatory mediators, reduced apoptosis and
contributed to efficient muscular regeneration of mdx
6
dystrophic mice . Therefore, we sought to isolate
molecules from the dichloromethane extract to test
for cholinergic neurotransmission experimental
models.
Results and Discussion
E. punicifolia was successively extracted at room
temperature with solvents of increasing polarity as
hexane, dichloromethane, and methanol. The
extracts were evaporated to dryness and the
residues stored at -20º C. Merck silica gel (230-400
mesh) was used for column chromatography. Merck
5554 Kieselgel 60 F254 sheets were used for thinlayer
chromatographic
(TLC)
analysis.
Dichloromethane extract was fractionated by column
chromatography, and 43 fractions were obtained.
Fractions 28-35 were pooled and it was observed
the formation of a precipitate named M1 that was
recovered and analysed. General experimental
procedures: Melting-point:Kofler block. Optical
rotation: in CHCl3 solution (Perkin-Elmer 241 MC
polarimeter). IR: in KBr (Perkin-Elmer Spectrum One
H
C
spectrophotometer). 1 and 13 NMR spectra (at
500 MHz and 125 MHz, respectively): in CD 3ODCDCl3 (9:1) solution at room temperature (Varian
36a Reunião Anual da Sociedade Brasileira de Química
H
C
SYSTEM 500 MHz spectrometer). 1 and 13 NMR
chemical shifts are reported with respect to the
H
CD3OD signals at  3.30 for 1 and at  49.00 for
C
13 . HRESIMS: Agilent 6520 Accurate-Mass QTOF
LC/MS apparatus. The constituent of M1 fraction
was ursane triterpene barbinervic acid (3α,19α,24
trihydroxyurs-12-en-28-oic acid). The identification
was supported by its physical (mp, [α]D) and
H
C
spectroscopic (IR, 1 and 13 NMR and mass
spectra) data, which were in complete agreement
with those reported in the literature (Takani et al.,
1977; Takahashi and Takani, 1978).
30
OH
29
12
11
1
2
9
4
5
H
2 4 23
H
8
7
6
H
13 17
14
26
10
3
HO
HO
25
19
18
HO
20
21
H
22
COOH
COOH
16 28
15
H
27
HO
HO
H
.
Figure 1. M1: barbinervic acid. (mp 295-298 ºC;
[α]D20 +18.3º (c 0.12, EtOH); C30H49O5m/z
489.3575)
Conclusion
Considering that triterpenoids are among molecules
responsible for some reported E. punicifolia effects,
it is imperative to perform experiments with
barbinervic
acid
to
test
for
cholinergic
neurotransmission models.
Acknowledgements
CIGS, Manaus, AM, Brazil for supplying the plant
and CAPES/DGU Program, FAPERJ and also
Fundación Carolina (España) for financial support.
___________________
1
Brito FA, et al. Braz. J. Med.Biol. Res.2007, 40:105–115
Bopp A. et al. Fundam.Clin.Pharmacol.2009, 23:501–507
Consolini and Sarubbio. J Ethnopharmacol.2002, 81:57–63
4
Grangeiro et al. J Ethnopharmacol.2006, 108:26–30
5
Pascual et al. Braz. J. Pharmacognosy2012, 22(1): 1-12
6
Leite et al. J. Cell Biochem.2010, 111:1652–1660
7
Takani et al. Chem. Pharm. Bull.1977, 25: 981-985
8
Takahashi and Takani, Chem. Pharm. Bull.1978, 26: 2689-2693
2
3