[生物医药论文精品]黑老虎的化学成分研究

黑老虎的化学成分研究A1A0A2A2A3A3A4A5A6A7A8A9A10KADSURACOCCINEAA1A11A12A13A14A15A16A17A18A19A20A21A22A23A24A25A26A27A28A29A30A16A17A14A15A15A31A32A33A34A35A36A12A13A37A38A39A40A28A13A41A42A6A15A31A43A44A11A12A45A35A16A17A46A47A48A49A20A50A51A52A7A8A9A53A15A31A43A447A54A55A11A56A57A58A59A60A61A62A63A12A45A35A32A15A64A48A49A6523OXO3,4SECO9HLANOST428,7DIEN3OICACIDA101A66A6723OXO3,4SECO9HLANOST428,7,24TRIEN3OICACIDA682A69A7025HYDROXY23OXO3,4SECO9HLANOST428,7DIEN3OICACIDA713A72A7325HYDROXY3,4SECO9HLANOST428,7,23TRIEN3OICACIDA744A75A7624RHYDROXY3,4SECO9HLANOST428,7,2526TRIEN3OICACIDA775A78A7924E3,4SECO8HLANOST428,911,24TRIEN3,26DIOICACIDA806A81A8223OXO8HLANOST911ENE3,4OLIDEA837A84A85A86A87A86A87A86A88A89A901A912A913A925A93A94A95A96A97A98A99HL60A100A101A102A103A104A105A91A106A107A108A88A89A901A95A96A100A101A102A103A109A104A105A110A111A85A112A113A114A115A116A117A118A119A120A121A122A123A119A124A125A126A127STUDIESONCHEMICALCONSTITUENTSOFKADSURACOCCINEAABSTRACTOBJECTIVETOSTUDYTHECHEMICALCONSTITUENTSOFTHEROOTSOFKADSURACOCCINEAMETHODSTHECOMPOUNDSWEREISOLATEDBYCHROMATOGRAPHYONSILICAGELTHEIRSTRUCTURESWEREIDENTIFIEDBYTHEIRPHYSICALANDSPECTRALDATARESULTSSEVENCOMPOUNDSWEREISOLATEDANDIDENTIFIEDAS23OXO3,4SECO9HLANOST428,7DIEN3OICACID1,23OXO3,4SECO9HLANOST428,7,24TRIEN3OICACID2,25HYDROXY23OXO3,4SECO9HLANOST428,7DIEN3OICACID3,25HYDROXY3,4SECO9HLANOST428,7,23TRIEN3OICACID4,24RHYDROXY3,4SECO9HLANOST428,7,2526TRIEN3OICACID5,24E3,4SECO8HLANOST428,911,24TRIEN3,26DIOICACID6AND23OXO8HLANOST911ENE3,4OLIDE7CONCLUSIONCOMPOUNDS1,2,3,AND5SHOWEDINHIBITORYACTIVITYAGAINSTHL60HUMANLEUKEMIACELLS,WHILECOMPOUND1WASTHEMOSTEFFECTIVEANTIPROLIFERATIVEAGENTKEYWORDSKADSURACOCCINEACHEMICALCONSTITUENTS,STRUCTUREIDENTIFICATION黑老虎为五味子科南五味子属植物KADSURACOCCINEALEMACSM的根及藤茎1。广泛分布于我国广东、广西、海南、贵州、四川等华南地区和西南地区。其性温，味辛、微苦，具有行气止痛、祛风活络、散瘀消肿之功效。黑老虎在民间常用于治疗胃、十二指肠溃疡、慢性胃炎、急性胃肠炎、风湿性关节炎、跌打肿痛、痛经、产后瘀血腹痛等症2。广西中医药研究所曾于1985年从南五味子属植物提取分离，研制成功的一种抗炎止痛药“祛风痛片”，祛风痛片对急性风湿性关节炎和慢性关节炎疗效显著，对肩周炎也有一定疗效3。为了阐明黑老虎的化学成分，为其开发利用奠定基础，本G7003对黑老虎根的化学成分G17839行了G17751G13007G13491的研究，从中分离G5483G20527G1022G7044的G13662G8623G14038G9931G3423G989G14832化G2524物。1仪器与材料G7097本YANACOMPS3G3423显微G10088G9869G8991定G1214G708G7422G7669G8503G709G727G7071G1821G8991定利用JASCODIP370PERKINELMER241MCG7071G1821G1214G727G7692G11925G1861G6403G1821G16901利用JEOLECP500G3423G7692G11925G1861G6403G1821G16901G1214G8991定G708TMSG1328G1881G7643G709G727HRFABMS利用JEOLJMS700G17148G16901G1214G8991定G727TLC和G7621G14406G16901用G11801G14026G708200G794300G11458G709G17153于G19750G4719海G8927化G5049G2390G727SEPHADEXLH20为PHARMACIA产G2709G727G7621G14406G16901G16809G2070G16280G7696G3355为分G7524G13443，其中所用G11719G8845G18286为中G8844G12255G7086090G263G709。黑老虎药G7460G17153于广西G11477南G4437G5078，G11013广西中医药研究所G17194G14550G12089G2115研究G2604G18504定为KADSURACOCCINEALEMACSM。2提取与分离黑老虎根G70810KGG709G12893G11874后，用95G1069G18267G3250G8981提取3G8437，G8611G84372G4579G7114，提取G9094G8999G13565后G5483G9036G141791340G。G4570G9036G14179G2164G17878G18339G14988G20323G8712G9163G5760，G1393G8437用G11719G8845G18286、G8707G1235、G1069G18252G1069G18243、G8503G981G18267G14807取，分G2047G3250G6922G2520G14807取G9354G2070G5483G11719G8845G18286G14807取物150G，G8707G1235G14807取物50G，G1069G18252G1069G18243G14807取物20G，G8503G981G18267G14807取物100G。G11719G8845G18286G14807取物G1209G11719G8845G18286G1069G18252G1069G18243G9163G2524G9354G2070G17839行G7811G5242G8939G14085，G7081007G709G8939G14085G5483化G2524物1G708110MGG709，G7081008G709G8939G14085G5483化G2524物2G708290MGG709，G70810015G709G8939G14085G5483化G2524物4G7088MGG709、5G7087MGG709、6G7086MGG709，G70810030G709G8939G14085G5483化G2524物3G70810MGG709。G8707G1235G14807取物G1209G11719G8845G18286G1005G18242为G8939G14085G2070，G70810015G709G8939G14085G5483化G2524物7G70822MGG709。3结构鉴定化G2524物1G726G7092G14406G19036G7242，MP148G794149C，20D368G708C0056，CHCL3G709。HRFABMSG13485G1998G1946分子离子G4804M/ZG7264573674MH，G6524G8991分子式为C30H49O3G708CALCD4573682FORMHG709。1HNMRG708CHCL3，500MHZG709高场区显示4G1022甲基单G4804G17148子信号082、093、107、185G708G25203H，SG709和3G1022甲基双G4804G17148子信号097G7083H，D，J58HZG709，092G7083H，D，J67HZG709和091G7083H，D，J64HZG709G72713CNMR中G13485G199830G1022碳信号，G6524断该化G2524物1为G989G14832类化G2524物。1HNMR中498G7081H，SG709和499G7081H，SG709及13CNMR中1497G708C4G709和1122G708C28G709，示结构中有末端双键存在，1769G708C3G709为羧基碳信号。结G2524EIMSM/ZG726383G7081566G709MCH2CH2CO2H4，并参考相关G7003献5，五味子科G989G14832G18252及G1881G18243成分的基本骨架为G13662G8623甾24Z烯26G18252G708LANOST24ZEN26OICACIDG709，表明存在3，4裂环G13662G8623甾428烯3G18252的结构。G4570化G2524物1的1HNMR、13CNMR信号通过HMQC和HMBCG17839行归属。烯氢信号532G7081H，D，J25HZG709分G2047与C5G708457G709和C6G708297G709存在远G12255相关，表明环G1881双键位于C7、C8位G727羰基碳信号2102分G2047与H22G708247和218G709及H24G708231和229G709存在远G12255相关，确定羰基连接在C23位。在化G2524物1的NOESYG16901中，H9G708264G709分G2047与H18G708082G709和H19G708093G709显示NOE相关，示H9为G3423。因此G6524断化G2524物1结构为23OXO3,4SECO9HLANOST428,7DIEN3OICACID，命名为SECOCOCCINICACIDA。化G2524物2G726G7092G14406G19036G7242，MP145G794146C，20D254G708C0032，CHCL3G709。HRFABMSG13485G1998G1946分子离子G4804M/ZG7264553539MHG708CALCD4553525G709，分子式G726C30H47O3。化G2524物2表现G1998与化G2524物1相似的NMRG16901图，其1HNMR、13CNMR信号通过HMQC和HMBCG17839行归属。化G2524物2比化G2524物1多G1998一组烯碳信号1238G708C24G709和1539G708C25G709G727另外化G2524物2还增G2164了2G1022双G4804甲基G17148子信号177G708CH326G709和222G708CH327G709，表明C24位存在双键。因此化G2524物2结构G6524断为23OXO3,4SECO9HLANOST428,7,24TRIEN3OICACID，命名为SECOCOCCINICACIDB。化G2524物3G726G7092G14406G19036G7242，MP181G794182C，20D282G708C0023，CHCL3G709。HRFABMSG13485G1998G1946分子离子G4804M/ZG7264953438MNAG708CALCD4953450G709，分子式G726C30H48O4。化G2524物3表现G1998与化G2524物1相似的NMRG16901图，其1HNMR、13CNMR信号通过HMQC和HMBCG17839行归属。化G2524物3比化G2524物1多G1998一G1022连氧碳信号695，且26、27位甲基G17148子信号G3355向低场位移至151。HMBCG16901显示，695与H24G708280G709、CH326G708151G709及CH327G708151G709存在相关，表明C25位连有一G1022羟基。因此化G2524物3的结构确定为25HYDROXY23OXO3,4SECO9HLANOST428,7DIEN3OICACID，命名为SECOCOCCINICACIDC。化G2524物4G726G7092G14406G19036G7242，MP183G794185C，20D311G708C0010，CHCL3G709。HRFABMSG13485G1998G1946分子离子G4804M/ZG7264573639MHG708CALCD4573682G709，分子式G726C30H49O3。化G2524物4表现G1998与化G2524物1相似的NMRG16901图，其1HNMR、13CNMR信号通过HMQC和HMBCG17839行归属。化G2524物4比化G2524物1多G1998一G1022连氧碳信号697及一组烯碳信号1246G708C23G709和1417G708C24G709。HMBCG16901显示，697与CH326G708155G709及CH327G708155G709存在相关，表明羟基连接在C25位G7271417与H22228，CH326及CH327存在远G12255相关，1246与H22存在相关，确定双键位于C23位。因此化G2524物4的结构确定为25HYDROXY3,4SECO9HLANOST428,7,23TRIEN3OICACID，命名为SECOCOCCINICACIDD。化G2524物5G726G7092G14406G19036G7242，MP161G794162C，20D209G708C0020，CHCL3G709。HRFABMSG13485G1998G1946分子离子G4804M/ZG7264793495MNAG708CALCD4793501G709，分子式G726C30H48O3。NMRG16901图显示化G2524物5与化G2524物1具有相同的母G7692，其1HNMR、13CNMR信号通过HMQC和HMBCG17839行归属。化G2524物5比化G2524物1多G1998一G1022连氧碳信号757及一组末端双键碳信号1495和1101。HMBCG16901显示，连氧碳信号757与H23G708193和169G709，H26G708529和498G709G1209及CH327G708192G709存在远G12255相关，表明C24位连有一G1022羟基G727烯碳信号1101与H24G708437G709、H27G708192G709的相关，确定结构中另一G1022末端双键位于C26位。C24的绝对构G3423G11013改良MOSHERS法确定，RMTPAG18243的1HNMRG16901与SMTPAG18243相比，H226和H327向高场位移，而H223和H120向低场位移。根据MOSHERS法的G16280则，在MTPA平面右侧的G17148子G13485G1998G8503的值，在MTPA平面左侧的G17148子G13485G1998负的值，因此C24的绝对构G3423为24R6。故化G2524物5结构确定为24RHYDROXY3,4SECO9HLANOST428,7,2526TRIEN3OICACID，命名为SECOCOCCINICACIDE。化G2524物6G726G7092G14406G19036G7242，MP111G794112，20D518G708C0022，CHCL3G709。HRFABMSG13485G1998G1946分子离子G4804M/ZG7264713472MHG708CALCDFOR4713474MHG709，分子式G726C30H46O4。化G2524物6与化G2524物1的NMRG16901图很相似，1HNMR中末端双键G17148子信号496G7081H，SG709和498G7081H，SG709及13CNMR中1141G708C28G709，1482G708C4G709和1768G708C3G709的碳信号，表明结构中存在3，4裂环G13662G8623甾428烯3G18252的结构7。G4570化G2524物6的1HNMR、13CNMR信号通过HMQC和HMBCG17839行归属。HMBCG16901显示，烯氢信号558与C10G708429G709，C12G708379G709及C13G708442G709存在远G12255相关，表明环G1881双键位于C9，C11位G7271425与H23G708234和217G709和H27G708213G709的远G12255相关表明侧链上C24，C25位存在双键G727羧基碳信号1707与H24G708721G709，CH327G708213G709存在相关，表明C26位连有一G1022羧基基团。NOESYG16901中CH327与H23存在NOE相关，故G4570C24构G3423确定为24EG727H8分G2047与H18、H19和H29存在NOE相关，示H8为G3423。因此化G2524物6结构确定为24E3,4SECO8HLANOST428,911,24TRIEN3,26DIOICACID，命名为SECOCOCCINICACIDF。化G2524物7G726G7092G14406G19036G7242，MP178G794179，20D802G708C0027，CHCL3G709。HRFABMSG13485G1998G1946分子离子G4804M/ZG7264573685MHG708CALCD4573682G709，分子式G726C30H48O3。1HNMRG16901中G13485G19985G1022单G4804甲基G17148子信号和3G1022双G4804甲基G17148子信号G72713CNMRG16901中显示30G1022碳信号，G6524断化G2524物7为G989G14832类化G2524物。NMRG16901图显示化G2524物7与化G2524物1具有相同的侧链结构，其1HNMR、13CNMR信号通过HMQC和HMBCG17839行归属。HMBCG16901显示，C4G708859G709与H28G708139G709、H29G708142G709和H5G708166G709的远G12255相关，G1209及C3G7081744G709与H1G708190G709和H2G708278G709的相关，表明A环存在G1881G18243结构G727烯氢信号526与C12G708376G709和C10G708422G709存在相关，G4570双键位置确定在C9，C11位。在化G2524物7的NOESYG16901中，H8分G2047与H18、H19和H28显示NOE相关，示H8为G3423。因此确定化G2524物7结构为23OXO8HLANOST911ENE3,4OLIDE，命名为COCCINILACTONEA。活性研究表明化G2524物1，2，3和5具有抑制白血病HL60细胞增殖G1328用，GI50分G2047为68136，133099，121237和421102M。A128A129A130A1311A132A133A134A135A136A137A138A1392005A140A141A142A143A144A145A146A147A132A134A135A148A149A150A151SA152A153A154A155A156A157A158A159A160A161A1622005,A163A164242A165A166A167A168A156A169A170A171A148A172A173A151MA152A153A154A174A175A176A177A159A160A161A162200121723A178A179A180A162A181A174A182A183A184A185A186A187A188A189A190A191A168A192A185A193A194JA167A195A168,1986,171094TAIT,AKAHORIA,SHINGUTTRITERPENOIDSFROMPORIACOCOSJPHYTOCHEMISTRY1991,30827965A196A158A197A162A198A199A200A162A201A202A203A204A205A206A207A168A208A209A210A167A211A212A213A214A215A216A217A218A219A220A221A222A223JA155A156A224A225A226A227A208,2001,1343636OHTANII,KUSUMIT,KASHMANY,ETALHIGHFIELDFTNMRAPPLICATIONOFMOSHERSMETHODTHEABSOLUTECONFIGURATIONSOFMARINETERPENOIDSJJAMCHEMSOC1991,1131140927KIMHJ,CHOIEH,LEEISTWOLANOSTANETRITERPENOIDSFROMABIESKOREANEJPHYTOCHEMISTRY,2004,65182545TABLE11HNMRDATA500MHZOFCOMPOUNDS17H,PYRIDINED5,JINHZH123456712031H,M1991H,M2031H,M1981H,M2021H,M1971H,M2031H,M1921H,M2031H,M1961H,M2762H,M1902H,M22572H,M2572H,M2562H,M2572H,M2572H,M2371H,M2111H,M2782H,M52261H,D,712261H,D,712251H,D,712261H,D,692261H,D,712191H,M1661H,M62381H,M2071H,M2391H,M2091H,M2381H,M2061H,M2401H,M2071H,M2401H,M2081H,M1921H,M1731H,M1542H,M75321H,D,255311H,D,285301H,D,305321H,D,355311H,D,281502H,M1872H,M82121H,M2141H,M92641H,M2641H,M2631H,M2651H,M2641H,M111691H,M1541H,M1681H,M1551H,M1681H,M1521H,M1532H,M1671H,M1551H,M5581H,D,555261H,D,58121811H,M1641H,M1811H,M1641H,M1801H,M1641H,M1771H,M1621H,M1841H,M1641H,M2081H,M1871H,M2091H,M1891H,M151561H,M1441H,M1541H,M1461H,M1541H,M1451H,M1541H,M1461H,M1551H,M1441H,M1362H,M1362H,M161901H,M1241H,M1931H,M1291H,M1901H,M1261H,M1962H,M1281H,M1971H,M1301H,M1522H,M1552H,M171531H,M1551H,M1531H,M1531H,M1521H,M1631H,M1661H,M180823H,S0823H,S0813H,S0813H,S0823H,S0703H,S0723H,S190933H,S0933H,S0933H,S0943H,S0943H,S1113H,S1163H,S202171H,M2181H,M2191H,M1521H,M1501H,M1471H,M2201H,M210973H,D,581013H,D,571013H,D,640953H,D,580963H,D,580973H,D,641013H,D,58222471H,M2181H,M2561H,M2181H,M2701H,M2401H,M2281H,M2251H,M1401H,M0971H,M1601H,M1201H,M2181H,M2481H,M235921H,M1931H,M1691H,M2341H,M2171H,M242311H,M2291H,M6181H,S2802H,D,715931H,M4371H,M7211H,M2312H,M252241H,M2241H,M260923H,D,671773H,S1513H,S1553H,S5291H,S4981H,S0943H,D,6427