Sesquiterpenoid bisabolane-jenis baru dari Curcuma domestica

 
Takahiro Ishii Hiroshi Matsuura b Kunimitsu Kaya b Charles Santhanaraju Vairappan , *
 
Laboratorium kimia produk alami, Institut biologi tropika dan Konservasi, Universiti
Malaysia Sabah, 88999 Kota Kinabalu, Sabah, Malaysia
 
b
Lulus sekolah kehidupan dan ilmu lingkungan, University of Tsukuba, 1-1-1 Tennodai,
Tsukuba, Ibaraki 305-8572, Jepang

1. subjek dan sumber

 
Kunyit adalah nama umum untuk ramuan tropis yang abadi, Curcuma domestica (syn.
Curcuma longa) keluarga jahe, Zingiberaceae. Secara luas didistribusikan di daerah tropis
dunia termasuk India, Indonesia dan Malaysia (Nagarathnam et al., 2010). Rimpang yang
telah secara tradisional dan secara luas digunakan sebagai aditif makanan dan zat pewarna
dalam banyak makanan dan kosmetik seluruh dunia (Jayaprakasha et al., 2005; Wang et al.,
2008). Di Malaysia, tanaman Zingiberaceae, Curcuma khususnya, secara ekstensif
dibudidayakan untuk pasar domestik dan internasional. Konstituen kimia mereka juga telah
menyelidiki karena pentingnya ekonomi dan biomedis (Vimala et al., 1999; Abas et al, 2005;
Malek et al., 2006). Kualitas Curcuma telah disarankan untuk berbeda dengan kualitas tanah
dan elevasi peternakan (Mridula dan darivera, 2001; Ishii et al., 2010). Oleh karena itu, kami
melakukan studi populasi untuk menyelidiki isi kimia dalam five populasi Curcuma dari
peternakan dataran rendah (di bawah 800 M) (Tamparuli, Kota Kinabalu, Sabah) dan delapan
populasi dari highland (di atas 800 M) (Tambunan, Sabah). Voucher spesimen yang disimpan
di koleksi BORNEENSIS Institute untuk biologi tropika dan Konservasi, Universiti Malaysia
Sabah, ((spesimen Tamparuli: GIN39888BOR-GIN39892BOR, Tambunan: GIN39893BOR-
GIN39900BOR)). Dalam penyelidikan ini, kami terisolasi sesquiterpenoid bisabolane-jenis
baru dari Curcuma tumbuh di highlands.

2. sebelumnya bekerja
 
Sebelumnya fitokimia penyelidikan pada genus Curcuma telah mengakibatkan isolasi
berbagai macam Metabolit sekunder, yang terutama mencakup sesquiterpenoids (Ohshiro et
al., 1990; Ia et al. 1998; Sofian et al., 2009; Sheeja dan Nair, 2010) dan diarylheptanoids
(Masuda et al., 1993; Taylor dan McDowell, 1992; Wang et al., 2008). Sampai saat ini,
jumlah bisabolane-jenis sesquiterpenoids telah terisolasi dari genus Curcuma (Kiso et al.,
1983; Uehara et al., 1989; Jayaprakasha et al, 2005; Zeng et al., 2007; Herebian et al., 2009 ).

3. Present work
 
Our investigation on the chemical constituents of the rhizome of C. domestica from
highland farms in Sabah, Malaysia led to the isolation of a new bisabolane-type
sesquiterpenoid, bisacurol B (1) together with seven known compounds, bisacurol (2)
(Uehara et al., 1989; Kreiser and Korner, 1999), E-a-atlantone (3) (Friesen and Blouin, 1996),
ar-turmerone (4) (Kamal et al., 2009), b-turmerone/curlone (5) (Kiso et al., 1983; Uehara et
al., 1989; Kreiser and Korner, 1999), curcumin (6) (Masuda et al., 1992),
demethoxycurcumin (7) (Masuda et al., 1992) and bisdemethoxycurcumin (8) (Masuda et al.,
1992). C. domestica from lowland farms only contained compounds 3–8. In this paper we
report the isolation and structural determination by spec-troscopic methods of new compound
1.
 
Fresh rhizome of C. domestica (500 g dry wt) was extracted in a Soxhlet apparatus with
EtOH. Concentration of the extract under reduced pressure afforded the crude extract (18.3
g). The EtOH fraction (670 mg) was chromatographed on a Si gel column using a
CHCl3/MeOH gradient solvent system of increasing polarity to yield 3 fractions. Fraction 1
(301 mg) eluted with CHCl3/MeOH (99:1) was subjected to reversed-phase HPLC (Luna 5m
Phenyl-Hexyl) with 70% MeCN to give compounds 3 (2.1 mg), 4 (2.4 mg) and 5 (2.8 mg). In
addition, fraction 2 (169 mg) eluted with CHCl 3/MeOH (95:5) was submitted to reversed-
phase HPLC (Luna 5m Phenyl-Hexyl) with 70% MeCN to yield a mixture of 1/2 (6.5 mg)
and 6 (6.8 mg). Fraction 3 (106 mg) eluted with CHCl 3/MeOH (9:1) was purified by repeated
preparative TLC with CHCl3/MeOH (98:2) to give compounds 7 (7.4 mg) and 8 (1.8 mg).

Senyawa 1 dan 2 sebanding dengan polaritas dan diperoleh sebagai campuran terpisahkan
dalam rasio ca. 1:1, seperti yang dihitung dari integrasi 1 H NMR. Upaya untuk memisahkan
campuran dengan berbagai tahapan stasioner dan mobile menggunakan HPLC dan mendaur
ulang sistem tidak berhasil. Oleh karena itu, penjelasan struktur dilakukan dalam campuran.
Campuran 1/2 diperoleh sebagai minyak berwarna, ½&28D 2.7 (c 0,45, CHCl 3 ). Rumus
molekul senyawa 1/2 bertekad untuk menjadi C15H24O oleh HR-ESI-TOFMS (m/z 203.1794
[Mþ H-H2O]þ (calcd. untuk C15H23, 203.1800), dalam menyimpulkan empat derajat dari
unsaturation. Campuran dipamerkan dua set sinyal di 1H dan 13C NMR spektrum. 1H dan 13C
NMR data dari salah satu pasangan (senyawa 2) dalam perjanjian yang baik dengan orang-
orang bisacurol senyawa, dikenal (Uehara et al., 1989; Kreiser dan Korner, 1999). Di sisi
lain, sinyal pola yang lain (senyawa 1) sangat menyerupai orang-orang 2 dan 13 C NMR
spektrum yang hampir identik dengan 2 kecuali siklik cincin. Oleh karena itu, senyawa 1
disarankan untuk menjadi bisabolane-jenis sesquiterpenoid serta analog dari 2.
 
Assignments were carried out based on 1H–1H COSY, HSQC and HMBC spectra data. 1H–
1
H COSY experiment revealed the sequences of the correlations depicted by the bold lines in
Fig. 2. Long-range COSY correlation between H-5 and H3-15 and HMBC correlations from
H3-15 to C-3, C-4 and C-5 indicated that the vinyl methyl group [d C 21.1 (q); dH 1.71 (3H, s)]
was attached to the olefinic carbon at C-4. HMBC correlations between H-2/C-4, H-3/C-5,
H-3/C-15 and H2-6/C-4 revealed the formation of a cyclic ring. In addition, HMBC cross-
peaks between H-10/C-12, H-10/C-13, H3-12/C-11, H3-12/C-13 and H3-13/ C-11 allowed us
to confirm the existence of isopropylidene moiety.
 
The NOESY experiment showed the limited correlations between H-1/H 2-6, H-3/H3-15, H-
5/H3-15, H-7/H3-14, H2-8/H-9, H2-8/H-10, H2-8/H3-14, H-9/H3-12, H-9/H3-14, H-10/H3-13,
and the relative stereochemistry of 1 could not be determined due to the absence of key
NOEs. However, judging from co-occurrence of 1 with 2 in the same specimen, 1 was
supposed to have the same configuration as that of 2. Therefore, based on the above data,
compound 1 was identified as 2-methyl-6-(40-methylcyclohexa-20,40-dien-10-yl)hept-2-en-4-
ol, namely bisacurol B. 1H NMR (600 MHz, CDCl3) d 5.78 (1H, brd, J ¼ 9.7 Hz, H-3), 5.62
(1H, dd, J ¼ 9.7, 3.0 Hz, H-2), 5.45 (1H, brs, H-5), 5.13 (1H, brd, J ¼ 7.6 Hz, H-10), 4.43
(1H, m, H-9), 2.29 (1H, m, H-1), 2.02 (2H, m, H-6), 1.73 (3H, s, H-13), 1.71 (3H, s, H-12),
1.71 (3H, s, H-15), 1.59 (1H, m, H-7), 1.49 (2H, m, H-8), 0.89 (3H, d, J ¼ 6.8 Hz, H-14). 13C
NMR (150 MHz, CDCl3) d 135.6 (C-11), 131.2 (C-4), 130.6 (C-2), 128.4 (C-3), 128.2 (C-
10), 120.3 (C-5), 67.3 (C-9), 42.1 (C-8), 38.3 (C-1), 33.3 (C-7), 25.9 (C-13), 24.5 (C-6), 21.1
(C-15), 18.3 (C-12), 17.1 (C-14). The structures of the compounds 2–8 were identified by the
comparison of their spectral data with those reported in literature. Fig. 1 represents the
structures of compounds 1–8.

4. chemotaxonomic makna
 
The major constituents of C. domestica are the diarylheptanoids (curcuminoids) and
sesquiterpenoids (Ohshiro et al., 1990; He et al., 1998; Herebian et al., 2009). The present
study reports the isolation of a new bisabolane-type sesquiterpenoid, bisacurol B (1), four
known sesquiterpenes 2–5 and three known curcuminoids 6–8 from the fresh rhizome of C.
domestica collected from highland farms. Compounds 3–8 have been previously reported
from the same species by Herebian et al. (2009), Raina et al. (2002) and, Taylor and
McDowell (1992). Bisacurol (2) has been previously obtained from Curcuma xanthorrhiza
(Uehara et al., 1989). To our knowledge, this is the second report on the presence of 2 in the
genus Curcuma. Our ongoing investigations with C. domestica from farms in Borneo have
shown the presence of chemical types that could have been contributed by factors such as
genetic variations, soil quality or microclimate (Ishii et al., 2010). Effect of soil type in the
quality of Curcuma was also investigated and reported by Mridula and Jayachandran (2001).
Based on this study, specimens obtained from highland farms contained bisabolane-type
sesquiterpenoids with a hydroxyl group at C-9. In contrast, the other specimens from lowland
farms only showed the presence of compounds 3–8. Therefore, occurrence of compounds 1
and 2 in C. domestica indicates that the bisabolane-type sesquiterpenoids with a hydroxyl
group at C-9 might be marker that can differentiate chemotypes within C. domestica and
might relate to the differences in the quality of chemotypes.