Separation Technology And Antitumor Activity Evaluation Of Components From Clausena Lenis

There are about 30 species of the genus Clausena?Rutaceae?in the Worldwide.It is mainly distributed in the tropical and subtropical areas of the eastern hemisphere.There are 10 species and 2 variants in China,from the southwest to Taiwan,mainly in Hainan,Guangdong,Guangxi,Yunnan and Taiwan.The plant of Clausena is not only a tropical fruit resource,but also a plant resource of medicinal.It has a long history of medicinal folk,which can be used for colds,stomach pain,rheumatoid arthritis,antispasmodic,anti-tumor,liver and lipid Multiple conditions.Studies have shown that the chemical components of Clausena are mainly alkaloids,coumarins,terpenes and other types of compounds.Pharmacological studies show that the genus Clausena plants have anti-tumor,antibacterial,antimalarial and anti-HIV and other activities.Clausena lenis is a plant of the genus Clausena?Rutaceae?which is mainly distributed in Hainan,southern Guangxi,and southern Yunnan in China.However,there are relatively few studies on the chemical constituents and pharmacological activities of Clausena lenis.In the previous research of this research group,the activity evaluation of 90%ethanol extract of Clausena lenis was conducted,and the results showed that it has obviously antitumor activity.Systematic research can not only provide scientific basis and material basis for the development of new antitumor drugs with high efficiency and low toxicity,but also lay a theoretical foundation for the rational use of medicinal plant resources of the genus Clausena.The dry stems of Clausena lenis was crushed,cold-extracted and extracted with90%ethanol,concentrated under reduced pressure,and extracted three times a week to obtain a total extract.Then,the chemical components in the extract were systematically separated and purified by separation techniques such as normal-phase silicagel column chromatography,ODS column chromatography,Sephadex LH-20gel column chromatography,and semi-preparative HPLC.We used the physicochemical properties,NMR,UV,IR,MS and other modern spectroscopy techniques to identify the structure of the isolated monomer compounds.56 monomer compounds were isolated and identified from 90%ethanol extracts of Clausena lenis,which were:clauselenin A?1?,clauselenin B?2?,clauselenin C?3?,6,8-dipreny lumbelliferone?4?,gravelliferone?5?,swietenocoumarin I?6?,3-?11-DimethylaU yl?-6-?2-hydroxy-3-methyl-3-buten-1-yl?-7-methoxycoumarin?7?,chalepin?8?,ang ustifolin?9?,9-?2-hydroxy-3-methoxy-3-methylbutoxy?bergapten?10?,byakangelicin?11?,tert-O-methylheraclenol?12?,pabularinone?13?,isogosferol?14?,heraclenol?15?,imperatorin?16?,trans decursidinol?17?,?+?-decursinol?18?,xanthyletin?19?,scataccanol?20?,isoangenomalin?21?,ma r m e s i n?22?,u mb e l l i f e r on e?23?,3-f or m y l c a r b a z o l e?24?,O-demethylmurrayanine?25?,murrayanine?26?,clausine Z?27?,clauszoline N?28?,clausine I?29?,2-Hydroxy-3-methyl-9H-carbazole?30?,glycozolidol?31?,methylcarbazole-3-carboxylate?32?,clausine E?33?,6-methoxycarbazole-3-carboxylate?34?,claulansine I?35?,2-methoxy-1-?3-methyl-buten-1-yl?-9H-carbazole-3-carbaldehyde?36?,clausehainanine A?37?,claulansine B?38?,claulansine D?39?,mafaicheenamine A?40?,mafaicheenamine B?41?,4-O-methyl-1-methyl-quinolin-2-one?42?,N-metilatanina?43?,skimmianine?44?,kokusaginine?45?,syringaresinol?46?,?+?sesamin?47?,sesamolin?48?,isololiolide?49?,blumenol B?50?,linolenic acid?51?,?-hydroxypropiovanillone?52?,syringaldehyde?53?,syringic acid?54?,coumarinic acid?55?,vanillic acid?56?.Among them,compounds 1-3 are new compounds,compounds 7?10?11?12?17?18?20?21?47?48?50?51?52?56 were isolated for the first time from the genus Clausena,except for compound 25,all other compounds were first separated from Clausena lenis.We used the modified MTT to evalu ated the antitumor activity of 40 compounds isolated from C.lenis.The evalua tion results showed that 13 monomer compounds have significant cytotoxic acti vity,the IC₅₀0 value was 0.63-17.22?M.The research provides the basis for the research of the anti-tumor activity of C.lenis,at the same time,it provides new resources for the search for highly effective and low-toxic anti-tumor drugs.