Академический Документы
Профессиональный Документы
Культура Документы
Fitoterapia
journal homepage: www.elsevier.com/locate/fitote
a r t i c l e i n f o a b s t r a c t
Article history: This study investigated the antioxidant and cytotoxic activity of the phenolics isolated from the
Received 6 July 2011 fruits of Livistona chinensis. Four new compounds, 1-{ω-isoferul[6- (4-hydroxybutyl)pentadecanoic
Received in revised form 19 September 2011 acid]}-glycerol (1), E-[6′-(5″-hydroxypentyl)tricosyl]-4-hydroxy-3-methoxycinnamate (2), 2-(3′-
Accepted 29 September 2011 hydroxy-5′-methoxyphenyl)-3-hydroxylmethyl-7-methoxy-2,3-dihydrobenzofuran-5- carboxylic
Available online 8 October 2011
acid (3), 7-hydroxy-5,4′-dimethoxy-2-arylbenzofuran (4), together with eleven known phenolics
(5–15), were isolated and identified. Among these compounds, 1–4, 5-O-caffeoylshikimic acid
Keywords: (5), caffeic acid (7), and 3-O-caffeoylshikimic acid (8) showed potent antioxidant activity. 1–5,
Livistona chinensis and 8 showed potent antiproliferative activities with IC50 values among 5–150 μM against HepG2
1-(ω-isoferuloylalkanoyl)-glycerol
human liver cancer, HL-60 human myeloid leukemia, K562 human myeloid leukemia, and CNE-1
Long-chain trans-ferulate
human nasopharyngeal carcinoma cell lines. On the basis of these findings, it could be proposed
Benzofuran
Antioxidant activity that the fruits of L. chinensis may serve as attractive mines of powerful anticancer and antioxidant
Antiproliferative activity agents for various purposes.
© 2011 Elsevier B.V. All rights reserved.
0367-326X/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.fitote.2011.09.020
X. Zeng et al. / Fitoterapia 83 (2012) 104–109 105
(HL-60, K562, HepG2, and CNE-1), and antioxidant assays for combined with preparative HPLC, yielding compounds 5
scavenging ability of DPPH and superoxide anion free radicals (12 mg), 10 (20 mg), 11 (25 mg), and 12 (15 mg), respec-
(O2−), were evaluated. tively. The CHCl3–MeOH (5:1) elution was subjected to an
octadecylsilanized silica gel (ODS) column, followed by a
2. Experimental preparative HPLC with 25% methanol (containing 0.1% CF3-
COOH, pH 3.0), yielding compound 3 (10 mg). The CHCl3–
2.1. General MeOH (3:1) elution was subjected to an ODS column,
followed by a preparative Rp-HPLC with 22% methanol
Optical rotations were measured using a JASCO P-1030 (containing 0.1% CF3COOH, pH 3.0), yielding compound
automatic digital polarimeter (Tokyo, Japan). NMR spectra 13 (10 mg). The structures of compounds 1–15 are shown
were recorded on a Bruker DPX-400 spectrometer using stan- in Fig. 1.
dard Bruker pulse programs. Chemical shifts were shown as 1-{ω-isoferul[6-(4-hydroxybutyl)pentadecanoic acid]}-glycerol
δ-values with reference to tetramethylsilane (TMS) as an inter- (1) was an amorphous powder; [α]25D +6.1 (c 0.20, CHCl3);
13
nal standard. GC–MS analysis was done using a Shimadzu GC- C NMR (CDCl3, 100 MHz) and 1H NMR (CDCl3, 400 MHz)
14A unit coupled with a GCMS-QP 2000 instrument (Tokyo, data, see Table 1; HRESIMS (positive ion mode) m/z [M+H]+
Japan). ESI-MS data were obtained on an Agilent 1200 579.3939 (calcd. 579.3931).
HPLC/6410B TripleQuad mass spectrometer (Santa Clara, CA), E - [6′ - (5″ - hydroxypentyl)tricosyl] - 4 - hydroxy - 3 -
and HR-ESIMS were measured on a Bruker APEX II mass spec- methoxycinnamate (2) was an amorphous powder; [α]25D-2.1
trometer (Bremen, Germany). Sephadex LH-20 (Pharmacia, (c 0.20, CHCl3); 13C NMR (CDCl3, 100 MHz) and 1H NMR
Sweden), silica gel (Qingdao Ocean Chemical Co., Ltd, Qingdao, (CDCl3, 400 MHz) data, see Table 1; HRESIMS (positive ion
China), and ODS (40–63 μm, Merck, Darmstadt, Germany) mode) m/z [M+ H]+ 603.4996 (calcd. 603.4991).
were used for column chromatography. TLC was carried out 2-(3′-hydroxy-5′-methoxyphenyl)-3-hydroxylmethyl-7-
on preparative silica gel 60 F254 and RP-18 F254 plates (Merck, methoxy-2,3-dihydrobenzofuran-5-carboxylic acid (3) was an
Darmstadt, Germany), and spots were visualized by spraying amorphous powder; [α] 25D + 8.2 (c 0.20, MeOH); 13C NMR
the plates with 15% H2SO4, and heating them at 105 °C. Prepar- (DMSO-d6, 100 MHz) and 1H NMR (DMSO-d6, 400 MHz) data,
ative HPLC was performed using an ODS column (XTerra®, see Table 2. HRESIMS (positive ion mode) m/z [M+ Na]+
19× 250 mm, 10 μm, Waters, Milford, MA). 369.0940 (calcd. 369.0945).
7-Hydroxy-5,4′-dimethoxy-2-arylbenzofuran (4) was yel-
2.2. Plant material low needles; 13C NMR (CDCl3, 100 MHz) and 1H NMR (CDCl3,
Table 1 Table 3
1
H NMR and 13C NMR data of compounds 1 and 2 in CDCl3 (δ in ppm, J in Hz). 1
H NMR, 13C NMR, 1H–1H COSY and HMBC data of compound 4 in CDCl3 (δ in
ppm, J in Hz).
Position Proton Carbon
1 13 1
Position H-NMR C- H–1H HMBC
1 2 1 2
NMR COSY
1 126.9 127.0
2 156.9
2 7.04 (s, 1H) 7.04 (d, 2, 1H) 109.1 109.3
3 6.79 (s, 1H) 100.4 C(2), C(3a), C(4),
3 147.7 147.9
C(5), C(7a), C(1′)
4 146.5 146.8
4 6.60 (d, 2.4, 1H) 94.8 H(6) C(3), C(5), C(6),
5 6.92 (d, 8.0, 1H) 6.92 (d, 7.6, 1H) 115.5 115.7
C(7), C(7a)
6 7.08 (br d, 8.6, 1H) 7.08 (dd, 7.6, 2, 1H) 122.9 123.0
5 145.5
7 7.62 (d, 16.0, 1H) 7.61 (d, 16.4, 1H) 144.5 144.6
6 6.43 (d, 2.0, 1H) 97.1 H(4) C(4), C(5), C(7), C(7a)
8 6.30 (d, 16.4, 1H) 6.29 (d, 16.4, 1H) 114.5 114.6
7 157.0
9 167.3 167.4
3a 131.0
\OCH3 3.93 (s, 3H) 3.93 (s, 3H) 55.8 55.9
7a 139.4
1′ 4.18 (m, 2H) 4.19 (t, 6.8, 2H) 64.9 65.0
1′ 123.6
\CH 2.32 (m, 1H) 2.34 (m) 34.0 34.1
2′ 7.73(d, 8.4, 1H) 126.9 H(3′) C(3′), C(4′), C(5′), C(6′)
\CH2b)c) 2.32 (m, 2H) 2.34 (m) 32.8 32.8
d) 3′ 6.89(d, 8.4, 1H) 115.9 H(2′) C(1′), C(4′), C(5′)
4′ 156.3
\Ca) 3.65 (t, 6.8, 2H) 173.7 63.1
5′ 6.89(d, 8.4, 1H) 115.9 H(6′) C(1′), C(3′), C(4′)
\CH2 1.25–1.64 (m, 2H) 1.25–1.69 (m) 28.8– 28.8–
6′ 7.73(d, 8.4, 1H) 126.9 H(5′) C(2′), C(3′), C(4′), C(5′)
31.8 31.9
5-OCH3 4.00 (s, 3H) 55.3 C(5)
\CH3 0.88 (t, 6.4, 3H) 0.88 (t, 6.4, 3H) 14.0 14.1
4′-OCH3 3.85 (s, 3H) 55.1 C(4′)
1″ 4.18 (m, 2H) 65.0
2″ 4.18 (m, 1H) 68.4
3″ 3.64 (t, 6.8, 1H) 63.1 CHCl3 was to give a white solid. GC–MS was performed to elu-
4.05 (t, 6.8, 1H) cidate the structure of the white solid. Also, the H2O fraction
a–d) Carbon groups in the structure of compounds 1 and 2 (see Fig. 1). was neutralized with dilute HCl and extracted with Et2O. The
Et2O extract was identified as ferulic acid by NMR analysis.
400 MHz) data, see Table 3. HRESIMS (positive ion mode) m/z 2.5. DPPH radical scavenging assay
[M+ H]+ 271.0967 (calcd. 271.0965).
The scavenging effects of the phenolics on DPPH radicals
2.4. Hydrolysis of compounds 1 and 2 were determined according to a previously reported method
[19]. DPPH (50 mg/L) was dissolved in MeOH. The samples
The hydrolysis reaction was determined according to the were dissolved in DMSO. The DPPH solution (995 μL) was
reported protocol [18]. Compound 1 or 2 (10 mg) was treated mixed with 5 μL of each of the samples. The mixture was shak-
with 5 mL 5% NaOH in EtOH. The mixture was refluxed for en and allowed to stand at room temperature in the dark for
10 h, and the solvent was removed by evaporation. The residue 20 min. The absorbance of the resulting solution was measured
was suspended in H2O and extracted with CHCl3. The dried spectrophotometrically at 517 nm.
Table 4 80
Antioxidant activities of the isolated compounds (1–9).
to large biomolecules, such as lipids, proteins, and DNA, result- [3] Zhao GP, Dai S, Chen ES. Dictionary of Traditional Chinese Medicine.
Shanghai: Shanghai Science and Technology Press; 2001. p. 2459–60.
ing in an increased risk for inflammatory diseases, cardiovascu- [4] Kaur G, Singh RP. Food Chem Toxicol 2008;46:2429–34.
lar disease, cancer, diabetes, Alzheimer's disease, cataracts, and [5] Li CY, Zeng YB, Peng F, Dai HF, Zheng YT. Chin Trad Herb Drugs 2008;39:
age-related functional decline [34–35]. The DPPH radical assay 1833–8.
[6] Wang H, Li A, Dong XP, Xu XY. J Chin Med Mat 2008;31:718–22.
was known to give reliable information concerning the antiox- [7] Chen Y, Lin XH, Li SG, Weng SM, Yao H. J Fujian Med Univ 2008;42:
idant ability of the tested compounds and extracts. At the same 93–5.
time, the superoxide anion free radical assay has been widely [8] Zhu YL, Chen SL, Peng LL, Tang L. Chem Bioeng 2007;24:35–7.
[9] Sartippour MR, Liu CH, Shao ZM, Go VL, Herber D, Nguyen M. Oncol Rep
used to evaluate the free radical scavenging ability. And this 2001;8:1355–7.
radical was generated by a chemical system composed of [10] Cheung S, Tai J. Oncol Rep 2005;14:1331–6.
PMS, NADH and oxygen. This assay was known to give reliable [11] Huang WC, Hsu RM, Chi LM, Leu YL, Chang YS, Yu JS. Cancer Lett
2007;248:137–46.
information concerning the antioxidant ability of the tested
[12] Chen P, Yang JS. Chin Trad Herb Drugs 2007;8:665–7.
compounds. The phenolics isolated from the fruits of cabbage [13] Chen P, Yang JS. Chin Pharm J 2008;43:1669–70.
palm showed potential free radical and superoxide anion free [14] Liu ZP, Cui JG, Liu HX. Cereal Oil Proc 2009;2:43–4.
radical (O2−) scavenging activities. [15] Tao Y, Yang SP, Zhang HY, Liao SG, Wei W, Yan W, et al. J Asian Nat Prod
Res 2009;11:243–9.
These results indicated the fruits of L. chinensis could be [16] Mala V, Dahot MU. Sci Int (Lahore) 1994;6:231–5.
served as cancer preventing agents, which were in accordance [17] Zeng XB, Qiu Q, Jiang CG, Jing YT, Qiu GF, He XJ. Fitoterpia 2011;82:
with their traditional uses. It can serve as potent anticancer and 609–14.
[18] Boonyaratavej S, Tantayanontha S, Kitchanachai P, Chaichantipyuth C,
antioxidant agents. The mechanisms of those compounds in Chittawong V, Miles DH. J Nat Prod 1992;55:1761–3.
cancer prevention are worthy of further investigation. [19] Yen GC, Chen HY. J Agric Food Chem 1995;43:27–32.
[20] Valentao P, Fernandes E, Carvalho F, Andrade PB, Seabra RM, Bastos
MD. Biol Pharm Bull 2002;25:1324–7.
Acknowledgments [21] He XJ, Liu RH. J Agric Food Chem 2007;55:4366–70.
[22] Kawanishi K, Hashimoto Y. Phytochemistry 1987;26:749–52.
This research was partly supported by the Start Fund of [23] Sy LK, Brown GD. Phytochemistry 1999;50:781–5.
[24] Yang XW, Zhao PJ, Ma YL, Xiao HT, Zuo YQ, He HP, et al. J Nat Prod
Wuhan University. The HR-ESIMS and optical rotations were 2007;70:521–5.
measured by Dr. Jinshan Tang, Institute of Traditional Chinese [25] Zhao XH, Chen DH, Si JY, Pan RL, Shen LG. Acta Pharmacol Sin 2002;37:
Medicine & Natural Products, Jinan University, China. The ESI- 535–8.
[26] Veit M, Weidner C, Strack D, Wray V, Witte L, Czygan FC. Phytochemistry
MS were kindly measured by Professor Naili Wang, Shenzhen 1992;31:3483–5.
Research Center of Traditional Chinese medicine & Natural [27] Chiang YM, Liu HK, Lo JM, Chien SC, Chan YF, Lee TH, et al. J Chin Chem
Products. Special thanks were given to Miss Lillian Nordahl, Soc 2003;50:161–6.
[28] Deyama T, Ikawa T, Kitagawa S, Nishibe S. Chem Pharm Bull 1987;35:
Department of Pharmacology at University of Cambridge, for 1785–9.
her kind help in manuscript preparation. [29] Ozawa T, Imagawa H. Agric Biol Chem 1988;52:595–7.
[30] Cussans NJ, Huckerby TN. Tetrahedron 1975;31:2719–26.
[31] Tsai IL, Hsieh CF, Duh CY, Chen IS. Phytochemistry 1996;43:1261–3.
References [32] Sandro M, Lauro ESB, Young GS, Patrick FJ, Chai HB, Eun JP, et al.
Phytochemistry 2002;59:635–41.
[1] Pei SJ, Cheng SY, Tong SJ. Flora of China. Beijing: Science Press; 1991. [33] Jayaprakasam B, Vanisree M, Zhang Y, Dewitt DL, Nair MG. J Agric Food
p. 25. Chem 2006;54:5375–81.
[2] Healthy Ministry of Guangzhou Force Logistics. Common Chinese Herbal [34] Ames BN, Gold LS. Mutat Res 1991;250:3–16.
Medicine Handbook. Beijing: People's Health Publishing House; 1969. [35] Ames BN, Shigenaga MK, Gold LS. Environ Health Perspect 1993;101:
p. 772–3. 35–44.