華鉤藤化學(xué)成分研究

劉明川，胡德禹，宋寶安，楊松，金林紅

貴州大學(xué)精細(xì)化工研究開發(fā)中心，教育部綠色農(nóng)藥與農(nóng)業(yè)生物工程重點(diǎn)實(shí)驗(yàn)室，貴陽550025

華鉤藤化學(xué)成分研究

劉明川，胡德禹，宋寶安，楊松*，金林紅

貴州大學(xué)精細(xì)化工研究開發(fā)中心，教育部綠色農(nóng)藥與農(nóng)業(yè)生物工程重點(diǎn)實(shí)驗(yàn)室，貴陽550025

從華鉤藤(Uncaria sinensis(Oliv．)Havil)的乙醇溶液中分離得到8個(gè)化合物，經(jīng)波譜學(xué)方法并與已知樣

品對(duì)照段鑒定為:β-谷甾醇(1)、胡蘿卜苷(2)、翅果定堿(3)、烏蘇酸(4)、丁香色原酮甙I(5)、美商陸甙A(6)、

東莨菪素(7)和烏蘇醛(8)。其中化合物5和6為首次從該植物中分離得到。

華鉤藤;化學(xué)成分

Introduction

Uncaria sinensis(Oliv．)Havil(Gouteng in Chinese) is widely distributed in southwestern of China，mainly in Guizhou，Guangxi，and there are 14 species in China［1-2］，among which 11 species are listed in Table 1．

Table 1 The distribution of the Uncaria Schreb．in China

U．scandens(Smith)HutchYunnan，Guangxi．U．yunnanensis K．C．HsiaYunnan U．laevigata Wal．ex G．DonYunnan U．homomalla MiqGuangxi

The whole plant of U．sinensis has been used for protecting liver，activating blood and antihypertensive for a long time［3，4］．Nowadays，more and more researchers have found that this plant's non-alkaloid parts also could be used as anticancer agents．Yang et al．［5］reported the triterpene compounds isolated from U．macrophylla Wall show anticancer bioactivity．Ursolic acid，3β-6β-23-trihydroxyurs-12-en-28-oic acid，showed proliferation inhibitory activity against U2OS with the inhibition rate 43．8%at the dosage of 25 mg/kg．d，while that of compound 3β-6β-23-trihydroxyurs-12-en-28-oic acid was 30．07%at 25 mg/kg．d．Mitsuo et al．［6］reported that the methanol extract from U．sinensis showed a suppressive effect on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen 3-amino-1，4-dimethyl-5H-pyrido［4，3b］indole(Trp-P-1)．Ursolic acid that was isolated from U．sinensis showed strong inhibiting effect and its ID50was 0．17 μmol/mL．

As far as we know，the chemical ingredients of U．sinensis were not reported yet．In this paper，the chemical constituents，especially the triterpene natural products from U．sinensis grown in Guizhou，were investigated．

Experimental

General

Melting point was determined on X4 melting point apparatus and the thermometer was uncorrected．1H and13C NMR spectra were recorded on a JOEL-ECX500 NMR instrument．LC-MS spectra were taken on an Agilent 1100/MSD．IR spectra were obtained on a SHIMADZU-IR Prestige-21，(KBr disks)．Silica gel(200-300 mesh，Qingdao Ocean Chemical Co．，Ltd，China) were used for column chromatographic separations．

Plant material

The plant of U．sinensis was purchased from Guizhou Province in August 2007，and identified by Professor Long Qingde(Guiyang Medical College)．The voucher specimens were deposited at Guiyang Medical College，Guiyang，China．

Extraction and isolation

The powdered U．sinensis(15 kg)was extracted with 85%EtOH(3×50 L)under reflux，6 h for the fist time，3h for the second time and 1h for the last time．After removal of the solvents by evaporation in vacco，the extract was suspended in water and then extracted with petroleum ether(10 L×3 times)，ethyl acetate (10 L×3times)and n-butanol(10 L×3 times)respectively．The petroleum ether(127 g)was subjected to silica gel column chromatography with petroleum ether:EtOAc(100∶1-1∶1)to give 14 fractions，then we use the silica gel column chromatography to separate those fractions．Finally we obtained 3 compounds 1(64 mg)，2(14 mg)，3(60 mg)．The ethyl acetate extract (346 g)was subjected to silica gel column chromatography with petroleum ether:CH3COCH3(100∶1-1∶1) to give 16 fractions．We used the silica gel column chromatography to separate those fractions again，and got five compounds from those fractions:4(120 mg)，5 (9 mg)，6(53 mg)，7(30 mg)and 8(4 mg)．

Structure Identification:

Compound 1White needles，mp.132-134℃，IR (KBr)cm-1:3429，2958，1462，1377，1053.The TLC of compound 1 was the same as β-sitosterol，so it was identified as β-sitosterol［4］.

Compound 2White powder，mp.298-301℃，IR (KBr)cm-1:3398，2958，1462，1374，1024.The TLC of compound 2 was the same as daucosterol，so it was identified as daucosterol［7］.

Compound 3White powder，mp.212-214℃，EI-MS m/z:368［M］+，337，353，223，180，130，69.IR (KBr)cm-1:3203，2945，1740，1676，1360.1H NMR (CDCl3，500 MHz)δ:0.88，1.42(3H，d，J=10 Hz，H-22)，1.60，1.71，2.00，2.17，2.47，3.21，3.29，3.60(3H，s，H-24)，4.34(1H，m，H-18)，7.6.87-7.26(4H，m，H-9，10，11，12)，7.41(1H，s)，8.18 (1H，s).13C NMR(CDCl3，125 MHz)δ:18.6(C-22)，30.1(C-14)，30.4(C-15)，34.8(C-20)，37.8 (C-6)，51.0(C-21)，53.5(C-24)，54.1(C-5)，56.9(C-7)，71.2(C-3)，72.1(C-19)，109.5(C-9)，109.8(C-16)，122.5(C-12)，124.5(C-11)，127.5(C-10)，133.7(C-8)，140.1(C-13)，154.9 (C-17)，167.63(C-23)，181.1(C-2).The IR，1H NMR，13C NMR and EI-MS data are in agreement with those of reported data of pteropodine，so we identify it as pteropodine［8］.

Compound 4White powder，mp.257-260℃.IR (KBr)cm-1:3450，2926，1695(C=O)，1456，1028，997.1H NMR:(500 MHz，CD3OH)δ:0.78(s，CH3)，0.82(s，CH3)，0.89(d，J=5 Hz，CH3)，1.16 (s，CH3)，3.2(1H，q，J=5.3 Hz，H-3)，5.2(1H，t，J=1.5 Hz，H-12).13C NMR(CDCl3，125 MHz)δ: 14.6(C-29)，14.7(C-30)，16.2(C-23)，16.5(C-24)，18.1(C-26)，20.1(C-25)，22.7(C-11)，23.0 (C-3)，24.0(C-21)，26.6(C-15)，27.4(C-7)，27.9(C-16)，30.4(C-6)，33.0(C-22)，36.7(C-12)，36.8(C-5)，38.5(C-13)，38.7(C-20)，39.0 (C-9)，39.1(C-19)，39.5(C-14)，41.9(C-10)，53.1(C-18)，55.4(C-9)，78.4(C-8)，125.6(C-2)，138.3(C-1)，180.1(C-28).The TLC of compound 4 was the same as ursolic acid，and IR，1HNMR，13C NMR data are in agreement with those of reported data of ursolic acid［9］.

Compound 5White powder，mp.245-248℃，EI-MS m/z:354［M］+，192，164，69，IR(KBr)cm-1:3429(-OH)，2926，1669(C=O)，1442，1091，677.1H NMR (CD3OD，500 MHz)δ:3.39～3.41(3H，m，H-2'，6')，3.49～3.52(1H，m，H-5')，3.8～3.9(2H，m，H-3'，4')，4.6～5.01(4H，m，glc-OH)，5.05(1H，d，J=10 Hz，H-1')，2.3(3H，s，H-9)，6.1(2H，s，H-5，6)，6.4(1H，s，H-7)，6.6(1H，s，H-2).13C NMR (CD3OD，125 MHz)δ:19.0(C-9)，61.0(C-6')，69.8(C-4')，73.3(C-2')，76.4(C-3')，77.0(C-5')，94.5(C-1')，107.9(C-7)，100.2(C-8a)，105.4(C-2)，99.6(C-5)，158.0(C-4a)，161.6(C-6)，163.4(C-8)，168.6(C-3)，182.8(C-1).The IR，1H NMR，13C NMR and EI-MS data are in agreement with those of reported data of eugenoside I［10，11］.

Compound 6White flake，mp.230-234℃，IR (KBr)cm-1:3400(-OH)，2972，1683(C=O)，1047.1H NMR(CD3OD，500 MHz)δ:3.80(2H，d，J =10 Hz，H-2')，3.26～3.35(2H，m，H-3'，4')，3.61 (1H，q，J=5 Hz，H-5')，4.32(1H，d，J=5 Hz，H-6')，5.57(1H，t，J=5 Hz，H-2)，4.1(3H，s，H-31)，4.3(2H，d，J=5 Hz，H-24)，2.27～0.72 is the hydrogen on the six-member rings.13C NMR(CD3OD，125 MHz)δ:177.8(C-28)，175.8(C-29)，132.7 (C-1)，128.9(C-2)，105.6(C-1')，88.4(C-8)，77.3(C-3')，76.4(C-2')，74.6(C-4')，71.0(C-5')，62.3(C-24)，55.6(C-31)，55.8(C-17)，55.6 (C-10)，54.2(C-4)，47.8(C-9)，46.7(C-14)，39.5(C-18)，39.0(C-19)，38.7(C-20)，38.5(C-5)，37.0(C-13)，36.7(C-12)，36.5(C-6)，36.2 (C-21)，30.0(C-15)，29.7(C-22)，27.5(C-16)，26.0(C-3)，24.5(C-7)，22.6(C-30)，20.63(C-27)，18.1(C-25)，17.1(C-11)，16.1(C-26)，15.9 (C-23).The IR，1H NMR，13C NMR data are in agreement with those of reported data of phytolaccoside A［12］.

Compound 7Yellow needles，mp.202-204℃，1H NMR(CD3COCD3，500 MHz)δ:8.75(1H，s，OH)，7.82(1H，d，J=10 Hz，H-4)，7.16(1H，s，H-5)，6.76(1H，s，H-8)，6.15(1H，d，J=10 Hz，H-3)，3.87(3H，s，OCH3).13C NMR(CD3COCD3，125 MHz)δ:160.4(C-2)，150.9(C-9)，150.3(C-7)， 145.1(C-4)，143.8(C-6)，112.5(C-10)，111.2 (C-3)，109.1(C-5)，102.8(C-8)，55.8(OCH3) .The IR，1H NMR，13C NMR data are in agreement with those of reported data of scopoletin［13］.

Compound 8White powder，mp.236-238℃，IR (KBr)cm-1:2926，1730.1H NMR(CDCl3，500 MHz) δ:0.77(s，CH3)，0.85(s，CH3)，0.87(d，J=6.5 Hz，CH3)，1.08(s.CH3)，3.2(s，br.3-H)，5.2 (1H，t，J=1.6 Hz，12-H).13C NMR(CDCl3，125 MHz)δ:15.5(C-29)，15.6(C-30)，17.0(C-23)，17.1(C-24)，18.3(C-26)，21.2(C-25)，23.3(C-11)，23.6(C-3)，24.2(C-21)，27.3(C-15)，28.0 (C-7)，28.2(C-16)，29.7(C-6)，31.0(C-22)，36.7(C-12)，37.0(C-5)，38.6(C-13)，38.8(C-20)，38.9(C-9)，39.1(C-19)，39.5(C-14)，41.9(C-10)，47.9(C-17)，47.6(C-4)，52.7(C-18)，〗55.3(C-9)，79.1(C-8)，125.9(C-2)，138.0(C-1)，207.1(C-28).The IR，1H and13C NMR data are in agreement with those of reported data of ursolic aldehyde［14］.

Acknowledgment This work is financially supported by Program for Excellent Scientist of Guizhou Province (［2005］0515)，Key Science and Technology Project of Guizhou Province(No.20076004)，and the National Key Technology R＆D Program(2006BAD07A12).

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Chemical Constituents from Uncaria sinensis(Oliv．)Havil

LIU Ming-chuan，HU De-yu，SONG Bao-an，YANG Song*，JIN Lin-hong
National-Guizhou Joint Laboratory of Biomass Comprehensive Utilization，Key Laboratory of Green Pesticide and Agricultural Bioengineering，Ministry of Education，Center for R＆D of Fine Chemicals，Guizhou University，Guiyang 550025，China

Eight compounds were isolated from Uncaria sinensis(Oliv．)Havil and their structures were identified by means of spectroscopic analysis as β-sitosterol(1)，daucosterol(2)，pteropodine(3)，ursolic acid(4)，eugenoside I (5)，phytolaccoside A(6)，scopoletin(7)，and ursolic aldehyde(8)．Compounds 5 and 6 were isolated from the U．sinensis for the first time．

Uncaria sinensis;chemical constituents

February 22，2010;Accepted May 11，2010

This work is financially supported by Program for Excellent Scientist of Guizhou Province(［2005］0515)，Key Science and Technology Project of Guizhou Province(No．20076004)，and the National Key Technology R＆D Program(2006BAD07A12)．

book=2011,ebook=23

R284．1;Q946．91

A

1001-6880(2011)06-1058-04

*Correspondence authorTel:86-851-8292171;E-mail:jhzx．msm@

gmail．com