茶藨子葉孔菌子實體(忍冬)的化學成分研究

李 聰,張永清,李 佳,邱麗麗

山東中醫藥大學,濟南 250355

茶藨子葉孔菌子實體(忍冬)的化學成分研究

李 聰,張永清＊,李 佳,邱麗麗

山東中醫藥大學,濟南 250355

從茶藨子葉孔菌(忍冬)中分離了 8個化合物,通過波譜法和理化性質分別鑒定為豆甾醇(1),二十八酸(2),β-谷甾醇 (3),麥角甾醇 (4),麥角甾醇過氧化物 (5),壬二酸 (6),煙酸 (7),原兒茶酸 (8)。化合物 1,3,6均為首次從該屬真菌中分到。

茶藨子葉孔菌 (忍冬);子實體;化學成分

Introduction

Phylloporia ribis(Lonicera japonicaThunb.),a fungi of the genusPhylloporiain the family Hymenochaetales,has been used as a traditional medicine for the treatment of pharyngitis,laryngitis and tonsillitis.The fruiting bodies ofPhylloporia ribis(Lonicera japonicaThunb.)revealed various biological activities including antitumor,immune-enhancing,anti-inflammatory activity and so on[1].In the course of a phytochemical study on the fruiting bodies ofPhylloporia ribis(Lonicera japonicaThunb.),eight compounds were isolated and identified as stigmasterol(1),n-octacosanoic acid (2),β-sitosterol(3),ergosterol(4),ergosterol-5,8-peroxide(5),azelaic acid(6),niacin(7)and protocatechuic acid(8).Compounds 1,3,6 were obtained from this genus for the first time.

Exper imental

General

NMR spectra were recorded on BrukerDRX-500 spectrometerswith T MS as internal standard.MS data were obtained on a Finnigan Trace mass spectrometer.Melting points were measured on an X-4 digital melting point apparatus and the temperature was uncorrected. Silica gel(100-200 mesh,200-300 mesh,Qing dao Marine Chemical Factory),Sephadex LH-20(Amersham Pharmacia Biotech),and ODS(50μm,Y MC Co.)were used for column chromatography.All the chemicals used in this studywere analytical grade.

PlantMaterial

Phylloporia ribis(Lonicera japonicaThunb.)was collected from Pingyi,Shandong Province,in August 2007,and was identified by Prof Zhang Xiao-Qing(Institute ofMicrobiology,China Academy of Science).A voucher spec imen was deposited in ShandongUniversity of Traditional ChineseMedicine.

Extraction and Isolation

The fruiting bodiesofPhylloporia ribis(Lonicera japonicaThunb.)(13 kg)was extracted three times(3h/ each)with 95%EtOH,and about 305 g of a residue was obtained after evaporating the solvents in vacuo. The residue was suspended in water and partitioned with petroleum ether,EtOAc,and n-BuOH,successively,to afford a petroleum ether extract(84 g),a EtOAc extract(47 g)and a n-BuOH extract(30 g).The petroleum ether fraction was subjected to silica gel column chromatography,and eluted with gradient of petroleum ether and EtOAc(from100:0 to 0:100,v/v)to yield 7 fractions(A-G).Fraction A was separated by column chromatography on silica gel with petroleum ether andMe2CO(100:1 to 1:100,v/v)and to afford Compound 1(20 mg).Compound 2(15 mg)was obtained from fraction B by column chromatography on silica gel(petroleum ether-Me2CO 100:1 to 1:100,v/ v).Fraction C was separated by column chromatography on silica gel(petroleum ether-Me2CO 100:1 to 1: 100,v/v)and compound 3(30 mg)was crystallized. Fraction D was separated by column chromatography on silica gel(petroleum ether-Me2CO 100:1 to 1:100,v/ v)to afford compound 4(10 mg).Compound 5(27 mg)was obtained by column chromatography on silica gel(petroleum ether-Me2CO 100:1 to 1:100,v/v) and was crystallized from(petroleum ether-Me2CO 7: 1)fraction and Sephadex LH-20(CHCl3-MeOH 1:1, v/v)to afford Compound 6(7 mg)from fraction E. The EtOAc fraction was subjected to silica gel column chromatography,and eluted with petroleum ether-Me2CO(5:1 to 1:1)and CHCl3-MeOH(20:1 to 1: 1,v/v)to yield 8 fractions(H-0)and Compound 7 (200 mg)was obtain from fraction I.Fraction L was separated by column chromatography on silica gel (CHCl3-MeOH 10:1 to 1:1,v/v),Sephadex LH-20 (MeOH)and ODS(MeOH-H2O 1:1,v/v)to afford compounds 8(7 mg).

Identification

Stigmasterol(1) Colorless needles(CHCl3),mp. 152-154℃.EI-MSm/s:412(M+),394,369,351, 327,301,273,255,199,145,95,69,55.The above data and Rf values on TLC are consistent with those of authentic sample.

N-octacosanoic acid(2) White powder(Me2CO), mp.78-80℃.EI-MSm/s:424(M+),396,368,73, 57,43;1H NMR(CDCl3,500MHZ)δ:0.85(3H,t,J=6.8 Hz),1.25(brs,CH2×n),1.65(2H,m), 2.30(2H,t,J=6.6 Hz).13C NMR (CDCl3,125 MHz)δ:14.1,22.7,24.7,31.9,33.7,29.4(CH2× n),179.7.These data were in agreement with those of n-octacosanoic acid[2].

β-sterol(3) Colorless needles(CHCl3),mp.139-140℃.EI-MSm/s:414(M+),396,381,329,303, 145.It showed a positive Libermann-Burchard reaction.It was identified asβ-sitosterol by comparision of the TLC behavior and the melting point with the authentic compound.

Ergosterol(4) Colorless needles(CHCl3),mp. 153-155℃.EI-MSm/s:396(M+),383,363,337, 271,253,227,171,105.1H NMR(CDCl3,500 MHz) δ:5.54(1H,bs,C6-H),5.36(1H,bs,C7-H),5.17 (2H,m,C22-H,C23-H),3.61(1H,m,C3-H),1.01 (3H,d,J=6.59 Hz,C21-Me),0.92(3H,s,C19-Me), 0.89(3H,d,J=6.87 Hz,C28-Me),0.82(3H,d,J= 6.32 Hz,C26-Me),0.78(3H,d,J=8.52 Hz,C27-Me),0.60(3H,s,C18-Me).13C NMR(CDCl3,125 MHz)δ:38.8(C-1),31.4(C-2),69.8(C-3),41.3 (C-4),140.9(C-5),119.5(C-6),116.9(C-7), 141.1(C-8),46.7(C-9),37.4(C-10),22.2(C-11),39.4(C-12),43.1(C-13),54.8(C-14),31.4 (C-15),19.0(C-16),56.0(C-17),40.9(C-18), 136.2(C-19),132.2(C-20),43.3(C-21),33.4 (C-22),16.2(C-23),11.9(C-24),21.1(C-25), 17.6(C-26),19.5(C-27),19.9(C-28).These data were in agreementwith those of ergosterol[3].

Ergosterol-5,8-peroxide (5) White needles (CHCl3),mp.169-173℃.EI-MSm/s:428(M+), 396,271;1H NMR(CDCl3,500 MHz)δ:0.81(3H, s,C18-Me),0.81(6H,d,J=7.0 Hz,C26,27-Me),0.88 (3H,s,C19-Me),0.90(3H,d,J=7.0 Hz,C28-Me), 1.00(3H,d,J=7.0 Hz,C21-Me),3.96(1H,m,H-3),5.12(1H,dd,J=15.5,8.0 Hz,C23-Me),5.24 (1H,dd,J=15.5,8.0 Hz,C22-Me),6.23(1H,d,J=8.5 Hz,H-6),6.49(1H,d,J=8.5 Hz,H-7).13C NMR(CDCl3,125 MHz)δ:36.9(C-1),30.1(C-2),66.4(C-3),34.7(C-4),79.4(C-5),130.7 (C-6),135.4(C-7),82.1(C-8),51.1(C-9),36.9 (C-10),20.9(C-11),39.3(C-12),44.5(C-13), 51.7(C-14),28.6(C-15),23.4(C-16),56.2(C-17),12.9(C-18),18.2(C-19),39.7(C-20),19.6 (C-21),132.3(C-22),135.2(C-23),42.8(C-24),33.1(C-25),19.9(C-26),20.6(C-27),17.6 (C-28).These datawere in agreementwith those of ergosterol-5,8-peroxide[4].

Azela ic acid(6) White needles(CHCl3),mp.107℃.EI-MSm/s:188(M+),171,152,124,83,41.1H NMR(CDCl3,500 MHz)δ:2.36(4H,t,J=7.2 Hz, H-2,8),1.64(4H,m,H-3,7),1.35(6H,m,H-4,5, 6).13C NMR(CDCl3,125 MHz)δ:179.5(C-1,9), 33.7(C-2,8),24.3(C-3,7),28.4(C-4,5,6). These data were identical to those of azelaic acid reported in the literature[5].

N iac in(7) White powders(CHCl3),mp.233-234℃.1H NMR(DMSO-d6,500MHZ)δ:7.54(1H,dd,J=7.9,4.8 Hz,5-H),8.26(1H,dd,J=7.9,1.9 Hz,4-H),8.78(1H,dd,J=4.8,1.9 Hz,6-H),9.06 (1H,d,J=1.2 Hz,2-H),13.41(1-H).13C NMR (DMSO-d6,125 MHz)δ:166.2(-COOH),153.4 (C-2),126.7(C-3),137.0(C-4),123.9(C-5), 150.3(C-6).These spectral data were in consistent with those of niacin[6].

Protocatechuic acid(8) White needles(CHCl3), mp.197-198℃.EI-MSm/s:154(M+),137,109, 81,63.1H NMR(DMSO-d6,500 MHz)δ:7.53(1H, d,J=2.0 Hz,H-2),7.47(1H,dd,J=8.5,2.0 Hz, H-6),6.90(1H,d,J=8.5 Hz,H-5).13C NMR (DMSO-d6,125 MHz)δ:123.0(C-1),117.2(C-2),145.2(C-3),150.3(C-4),115.4(C-5),123.3 (C-6),167.3(-COOH).These data were in agreementwith those of protocatechuic acid[7].

1 Xu DY(徐大勇),Shi JH(時建華),WangWL(王維煉),et al.The ingredient examination and phar macology research of honey-suckle stem mushroom.J L inyiM ed sch(臨沂醫專學報),1997,19:261-264.

2 Fan MS(范明松),Ye G(葉冠),Huang CG(黃成鋼). Studies on chemical constituents ofFicus pum ilaL..Chin Tradit Herb D rug(中草藥),2005,36:984-986.

3 Zhang XQ(張曉琦),Yin ZQ(殷志琦),Ye WC(葉文才). Chemical constituents from fruiting bodies ofGanoder m a lucidum.Chin Tradit Herb D rugs(中草藥),2005,36:1601-1603.

4 Li XJ(鸝皆秀),Li J(李進),Xu LZ(徐麗珍),et al.Studies on chemical constituents ofCordyceps sinensis(Berk) Sacc.Chin Phar m J(中國藥學雜志),2003,38:499-501.

5 Guo LN(郭麗娜),JiangLM(江黎明),Wu LJ(吳立軍),et al.Studies on the chemical constituents of the stems and leaves ofAcanthonanax sessiliflorusSeem.J Shenyang Phar m Univ(沈陽藥科大學學報),2002,19:180-182.

6 LiW(李薇),Chen FK(陳發奎),Yin XW(尹相武),et al. Chemical constituents ofFolium isatidis.J Shenyang Phar m Univ(沈陽藥科大學學報),2005,22:15-17.

7 Yin T(尹婷),Liu H(劉樺),Wang F(王邠),et al.Chemical constituents fromSpatholobus sinensis.Acta Phar m Sin(藥學學報),2008,43:67-70.

Chem ical Constituents from Fruiting Bodies of Phylloporia ribis(Lonicera japonicaThunb.)

L ICong,ZHANG Yong-qing＊,L IJia,Q IU Li-li
Shandong University of Traditional ChineseM edicine,Jinan 250355,China

Eight compoundswere isolated from the fruiting bodies ofPhylloporia ribis(Lonicera japonicaThunb.).On the basis of spectra data and physicochemical properties,they were identified as stigmasterol(1),n-octacosanoic acid (2),β-sterol(3),ergosterol(4),ergosterol-5,8-peroxide(5),azelaic acid(6),niacin(7),protocatechuic acid(8). Compounds 1,3,6 were obtained from this genus for the first t ime.

Phylloporia ribis(Lonicera japonicaThunb.);fruiting bodies;chemical constituents

October 13,2008;December 10,2008 Foundation Item:This work was finantially supported by the Qualitycontrolling Standard of Chinese Medicinal Materials and Preparation Technique of Stand Sample of ChineseMedicinal(2006GG1109078).

R284.1;Q946.91

A

1001-6880(2010)03-0422-03

＊Corresponding authorTel:86-531-89628085;E-mail:zyq622003@ 126.com