瑞香狼毒根中的一个新化合物

杨彩霞，雷 蕾，王小亮，张正凯

1 西北师范大学化学化工学院;2甘肃省高分子材料重点实验室;3生态环境相关高分子材料教育部重点实验室，兰州 730070

Introduction

Stellera chamaejasme L.is one species of the genus Stellera，belonging to family Thymelaeaceae.In China，this plant is mainly distributed in northwest and northeast areas，including Inner Mongolia，Qinghai，Gansu，Tibet provinces［1］.The dried roots of S.chamaejasme L.is a well-known traditional Chinese medicine.It is often used as expectorant，anti-inflammation，detoxification，apocatastasis agent and also used for the treatment of furuncle，carbuncle and ulcers［2，3］.Zhou L et al［4］.and Gong XX et al［5］.reported the antibacterial，antivirus，anticancer activity and bactericidal effect of compounds from the roots of S.chamaejasme L..Previous phytochemical investigation on S.chamaejasme L.had resulted in the isolation and identification of a variety of secondary metabolites，which mainly included coumarins，flavonoids，lignans and phenylpropanoid glycosides［3-6］.Our laboratory had recently reported a number of coumarins and flavonoids from the roots of S.chamaejasme L.［7］.As a continuation of our phytochemical investigation，the present study reported another 7 compounds of this plant.The structures of these compounds were identified as 3-butyryl-4-amino ethylcinnamate(1)，ferulic acid(2)，vanillic acid(3)，daphneticin(4)，5＇-demethoxy daphneticin(5)，3＇-hydroxy-4＇-O-β-D-glucopyranoside flavone(6)and 3-methoxy-4-O-β-D-gluco-benzoic acid(7)by comparing physicochemical properties and NMR data with published literatures.Among them，compounds 1-3，6-7 were reported from this species for the first time，compound 1 is a new compound.

Materials and Reagents

The roots of S.chamaejasme L.were collected in August 2008 from Huining county of Gansu province，China.It was identified by Professor CHEN Xue-lin(College of Life Science，Northwest Normal University).A voucher specimen(No.200807)was deposited in the College of Chemistry and Chemical Engineering，Northwest Normal University.

1H NMR and13C NMR spectral data were recorded on a Bruker-DRX-400 FT NMR spectrometer(400 MHz for1H NMR and 100 MHz for13C NMR)with tetramethylsilane(TMS)as internal standard;Electron ionization mass spectrometry(EI-MS)and HREI-MS spectral data were acquired on a Bruker APEX II;Silica gel(200-300，300-400 mesh)and silica gel GF254(10-40 μm)were purchased from Qingdao Hai Yang Chemical Group Company，Shandong;Sephadex LH-20 gel were used for column chromatography;Spots were detected on TLC under UV lamp or by heating after spraying with 5% H2SO4in C2H5OH(v/v).

Extraction and Isolation

The powder of dried roots(5 kg)of S.chamaejasme L.was exhaustively extracted with 90% ethanol 3 times under reflux.The combined extract was evaporated under reduced pressure to yield a syrupy residue(670 g).The residue was suspended in water and extracted with petroleum ether(PE，60-90 °C)，CHCl3，EtOAc and n-BuOH，successively.The PE fraction(71 g)，CHCl3fraction(26 g)，EtOAc fraction(125 g)and n-BuOH fraction(54 g)were yielded，respectively.The CHCl3fraction was chromatographed on silica gel column using gradient elution with PE/CHCl3(50∶1 to 1∶1)to yield 6 fractions(Fr.1-Fr.6).Fr.5 was sequentially separated on silica gel column eluted with CHCl3/CH3COCH3(30∶1 to 1∶1)to yield 2(20.9 mg)and 3(9.0 mg).The EtOAc fraction was subjected to silica gel column，gradiently eluted with CHCl3/EtOAc(50∶1 to 5∶1)，18 fractions were obtained.Fr.1 was retreated on silica gel column and eluted with CHCl3/EtOAc(20∶1 to 10∶1)to yield 4(23.3 mg)and 5(12 mg).The n-BuOH fraction was subjected to silica gel column using a gradient of EtOAc/MeOH(30∶1 to 1∶1)，4 fractions were obtained.Fr.1 was isolated and purified in combination of silica gel and Sephadex LH-20 column to yeild 1(15 mg).By the same method，compound 6(14 mg)and 7(9 mg)were obtained from Fr.2 eluted with CHCl2/MeOH(30∶1 to 1∶1).

Structural elucidation

Compound 1 was obtained as white crystal with a molecular formula of C15H19O3N deduced from its positive HREI-MS data(［M］+at m/z:261.1357，calc.261.1360).The IR(KBr)spectrum of 1 revealed absorption bands of amino(3480，3325 cm-1)，carbonyl(1688 cm-1)，aromatic ring(1603，1510，1444 cm-1)and double bond(1661 cm-1).The1H NMR spectrum of 1(Table 1)indicated the signals of a 1，3，4-trisubstituted benzene ring at δ 6.88，7.21(each 1H，d，J=8.0 Hz)and 7.61(1H，s)，a trans-double bond at δ 7.58 and 6.27(each 1H，d，J=16.0 Hz)，which suggested the presence of phenylpropanoid moiety［8］.Besides，conspicuous signals of an ethyoxyl group at δ 4.19(2H，q，J=7.6 Hz)and 1.35(3H，t，J=7.6 Hz)，an active hydrogen at 3.89(which can be exchanged by D2O)were also observed.The13C NMR and DEPT data of 1(Table 1)contained 15 signals，including 10 sp2carbons and 5 sp3carbons，which revealed two carbonyl groups at δC167.9(s)and 196.9(s)，a double bond carbon at 144.8(d)，114.4(d)，an aromatic carbons at 124.6(s)，127.8(d)，126.2(s)，145.7(s)，115.5(d)，132.4(d)and an ethyoxyl carbons at 61.4(t)，14.5(q).The presence of a butyryl group was deduced from the signals of δH0.87(t，3H)，1.53(m，2H)，2.88(t，2H)and δC14.1，18.7，40.3 and 196.9.Considering the molecular formula of 1，an ethyoxyl and an amino were presented in 1，in addition to the phenylpropanoid moiety and an butyryl group.The configuration between C-7 and C-8 was trans inferred from the coupling constant(16.0 Hz).The location of butyryl group was assigned by HMBC correlations of δH7.61(H-2)with δC196.9(C-1＇)and δH2.88(H-2＇)with δC196.9(C-1＇)(Fig.1).Besides，the observed correlations of δH7.58(H-7)with δC124.6(C-1)，δH6.27(H-8)with δC167.9(C-9)and δH4.19(H-10)with δC167.9(C-9)indicated that 1 had an acryloyl group located at C-1.Analysis and comparison of the1H-1H COSY and HMBC spectra supported the1H NMR and13C NMR assignments(Table 1)and further suggested that compound 1 was a derivative of phenylpropanoid containing a nitrogen atom.All spectra data of 1 were in agreement with the structure as shown in Fig.1.Therefore，the structure of 1 was established as 3-butyryl-4-amino ethylcinnamate.

Table 1 1H NMR(400 MHz)and13C NMR(100 MHz)data of 1(in CD3OD-d4，δ ppm，J in Hz)

Compound 2white crystal，C10H10O4，EI-MS m/z:194［M］+;1H NMR(400 MHz，CD3OD)δ:7.19(1H，d，J=2.0 Hz，H-2)，7.08(1H，dd，J=2.0，8.0 Hz，H-5)，6.83(1H，d，J=8.0 Hz，H-6)，7.63(1H，d，J=16.0 Hz，H-7)，6.34(1H，d，J=16.0 Hz，H-8)，3.91(3H，s，3-OCH3);13C NMR(100 MHz，CD3OD)δ:171.2(COOH)，127.8(C-1)，111.7(C-2)，150.6(C-3)，149.3(C-4)，116.7(C-5)，124.2(C-6)，146.8(C-7)，116.2(C-8)，55.9(3-OCH3).These spectral data were identical with those of ferulic acid［9］.

Compound 3white powder，C8H8O4，EI-MS m/z:168［M］+;1H NMR(400 MHz，DMSO-d6)δ:7.43(1H，d，J=1.6 Hz，H-2)，6.84(1H，d，J=8.0 Hz，H-5)，7.44(1H，dd，J=1.6，8.0 Hz，H-6)，12.47(1H，brs，-COOH)，9.80(1H，brs，4-OH)，3.81(3H，s，3-OCH3);13C NMR(100 MHz，DMSO-d6)δ:167.2(-COOH)，121.7(C-1)，115.1(C-2)，147.4(C-3)，149.8(C-4)，112.7(C-5)，123.6(C-6)，56.3(3-OCH3).The above spectral data were identical with those of vanillic acid［10］.

Compound 4colorless needles，C19H12O7，EI-ME m/z:352［M］+;1H NMR(400 MHz，DMSO-d6):6.34(1H，d，J=9.6 Hz，H-3)，8.00(1H，d，J=9.6 Hz，H-4)，7.20(1H，d，J=8.6 Hz，H-5)，6.96(1H，d，J=8.6 Hz，H-6)，6.76(2H，s，H-2＇，6＇)，4.43(1H，d，J=7.8 Hz，H-7＇)，4.34(1H，m，H-8＇)，3.40，3.68(each 1H，m，H-9＇)，3.81(6H，s，2×OCH3)，8.54(1H，s，4＇-OH)，4.36(1H，s，9＇-OH);13C NMR(100 MHz，DMSO-d6)δ:160.3(C-2)，113.5(C-3)，143.5(C-4)，112.0(C-5)，113.2(C-6)，147.7(C-7)，138.4(C-8)，149.2(C-9)，113.6(C-10)，126.5(C-1＇)，106.1(C-2＇)，149.3(C-3＇)，132.2(C-4＇)，149.3(C-5＇)，106.1(C-6＇)，77.2(C-7＇)，78.5(C-8＇)，60.5(C-9＇)，56.7(OCH3).The above spectral data were identical with those of daphneticin［11］.

Compound 5yellow powder，C19H16O7，EI-MS m/z:356［M］+;1H NMR(400 MHz，DMSO-d6)δ:6.32(1H，d，J=10.0 Hz，H-3)，7.98(1H，d，J=10.0 Hz，H-4)，7.19(1H，d，J=8.0 Hz，H-5)，6.95(1H，d，J=8.0 Hz，H-6)，7.03(1H，d，J=2.0 Hz，H-2＇)，6.81(1H，d，J=8.0 Hz，H-5＇)，6.88(1H，dd，J=8.0，2.0 Hz，H-6＇)，3.38，3.70(1H，m，H-9＇)，3.80(3H，s，-OCH3)，4.31(1H，m，H-8＇)，9.19(1H，s，4＇-OH)，4.45(1H，s，9＇-OH);13C NMR(100 MHz，DMSO-d6)δ:160.2(C-2)，112.5(C-3)，144.6(C-4)，119.8(C-5)，113.2(C-6)，146.8(C-7)，131.0(C-8)，143.1(C-9)，112.9(C-10)，126.5(C-1＇)，112.0(C-2＇)，146.9(C-3＇)，147.5(C-4＇)，115.3(C-5＇)，120.5(C-6＇)，76.5(C-7＇)，78.1(C-8＇)，59.9(C-9＇)，55.9(-OCH3).The above spectral data were identical with those of 5＇-demethoxy daphneticin［11］.

Compound 6colorless needles，C21H20O9，EI-MS m/z(neg.):415［M-H］-;1H NMR(400 MHz，DMSOd6)δ:6.89(1H，s，H-3)，8.02(1H，d，J=8.0 Hz，H-5)，7.48(1H，t，J=7.0 Hz，H-6)，7.80(1H，m，H-7)，7.79(1H，m，H-8)，7.55(1H，d，J=2.0 Hz，H-2＇)，7.24(1H，t，J=8.5 Hz，H-5＇)，7.55(1H，t，J=8.5 Hz，H-6＇)，9.10(1H，brs，3＇-OH)，4.85(1H，d，J=7.2 Hz，H-1＇＇)，3.15-3.73(6H，protons of glycoside);13C NMR(100 MHz，DMSO-d6)δ:162.5(C-2)，105.7(C-3)，177.2(C-4)，125.6(C-5)，134.4(C-6)，118.7(C-7)，124.6(C-8)，156.9(C-9)，124.9(C-10)，123.5(C-1＇)，113.5(C-2＇)，146.9(C-3＇)，148.5(C-4＇)，115.8(C-5＇)，118.7(C-6＇)，101.3(C-1＇＇)，73.3(C-2＇＇)，77.4(C-3＇＇)，69.7(C-4＇＇)，75.8(C-5＇＇)，60.8(C-6＇＇).These spectral data were identical with those of 3＇-hydroxy-4＇-O-β-D-glucopyranoside flavone［12］.

Compound 7white powder，C14H18O9，EI-MS m/z:330［M］+;1H NMR(400 MHz，DMSO-d6)δ:7.74(1H，brs，H-2)，7.48(1H，d，J=8.4 Hz，H-6)，7.14(1H，d，J=8.4 Hz，H-5)，5.01(1H，d，J=6.0 Hz，H-1＇)，3.66(1H，d，J=12.0 Hz H-6＇a)，3.45(1H，dd，J=5.6，12.0 Hz，H-6＇b)，3.35(1H，m，H-2＇)，3.18(1H，m，H-4＇)，3.35(1H，m，H-5＇)，3.80(3H，s，3-OCH3).These spectral data were identical with those of 3-methoxy-4-O-β-D-gluco-benzoic acid［13］.

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