Studies On The Chemical Components And Biological Activities Of Hemsleya Pengxianensis Var. Jinfushanensis

H.pengxianensis var.jinfushanensis belongs to Cucurbitaceae Hemsleya and mainly distributed in Yunnan,Guizhou,Sichuan,and Chongqing provinces of China.It is used for the treatment of enteritis,upper respiratory tract infection,bronchitis,bacterial dysentery,urinary tract infection and other diseases.Modern research revealed that H.pengxianensis var.jinfushanensis is rich in cucurbitane triterpenes and glycosides,lignans,organic acids and other constitutes.In order to comprehensively study the chemical constituents of H.pengxianensis var.jinfushanensis and to reveal the biological activities of chemical compositions and elucidate the therapeutic material basis of this herbal medicine,isolation of the chemical compositions of H.pengxianensis var.Jinfushanensis were conducted.Anticancer activities of the isolated compounds were screened in vitro and the UPLC MS/MS method for the determination of nine cucurbitane triterpenes and glycosides in H.pengxianensis var.jinfushanensis was established.The results of the current thesis established the foundation to make better use of H.pengxianensis var.Jinfushanensis and the detail conclusions were as follows:1.The investigations of ethyl acetate and n-butanol extracts of H.pengxianensis var.jinfushanensis using various chromatographic methods resulted in the isolation of 72 compounds including 70 cucurbitane triterpenes and glycosides,1 lignan and 1 organic acid.Their structures were identified on the basis of UV,IR,ESI-MS,HR-ESI-MS and NMR(1H-NMR,13C-NMR,1H-1H COSY,HSQC,HMBC,NOESY)spectra.Among them,30 compounds were identified as new compounds and 1 compound was obtained from the genus Hemsleya for the first time.All the new compounds were cucurbitane triterpenes and glycosides and named as hemsleyaoside A(Al),hemsleyaoside A1(A2),hemsleyaoside B(A3),hemsleyaoside C(A4),hemsleyaoside D(A5),hemsleyaoside E(A6),hemsleyaoside Ei(A7),hemsleyaoside E2(A8),hemsleyaoside E3(A9),hemsleyaoside F(A10),hemsleyaoside F1(A11),hemsleyaoside F2(A12),hemsleyacin A(A13),hemsleyaoside G(A 14),hemsleyaoside H(A 15),hemsleyacin B(A 16),hemsleyacin C(A 17),hemsleyacin D(A 18),hemsleyaoside I(A 19),hemsleyaoside J(A20),hemsleyacin E(A21),hemsleyacin E1(A22),hemsleyacin F(A23),hemsleyacin G(A24),hemsleyacin I(A25),hemsleyacin J(A26),hemsleyacin K(A27),hemsleyacin L(A28),hemsleyacin M(A29)and hemsleyacin N(A30).In addition,a series of 16,23-pyran ring cucurbitane triterpenes and glycosides,7-carbonyl substituted cucurbitane triterpenes and glycosides were also isolated from H.pengxianensis var.jinfushanensis and the ?,?-unsaturated ketone(27-demethyl)with side chain of triterpenoid saponin was isolated for the first time.2.Based on above-mentioned studies,we summarized the spectral regularities of the cucurbitane triterpenes and glycosides on the basis of the isolated compounds and the reported constituents from genus Hemsleya,and provide reference for rapiding identification of cucurbitane triterpenes and glycosides.3.On the basis of chemical research,MTT method was used to evaluate anticancer activities of the new compounds and some known compounds in vitro.The results showed that hemsleyaoside F2,hemsleyacin N,hemslecin A,hemslecin B,hemslepenside A,hemsleyacin B,hemsleyacin K,hemslecins G,cuburbitacin F and 16-O-acetyl-cucurbitacin F have different degrees of inhibition on Hela Cell proliferation activities with the IC50 less than 10 ?g/ml.Among them,hemslepenside A had the strongest inhibitory effect on Hela cell proliferation with an IC50 0.89 ?g/ml.Besides,the structure-activity relationships of cucurbitane triterpenes and their glycosides was also discussed.4.The UPLC-MS/MS method is efficient,sensitive and suitable for the determination of multi-index contents in H.pengxianensis var.jinfushanensis and preparations,which lays the foundation for quality control and evaluation of medicinal materials and their preparations.The contents of hemslecin A,hemslecin B,hemslecins G,xuedanencins B,delavanoside D,hemsleyaoside A,hemsleyacin N,cucuibitacin F and jinfushanoside C were 25.87,3.48,0.34,0.10,0.34,9.09,0.04,0.06,0.55 mg/g,respectively.The developed UPLC-MS/MS method could be reliably applied as a quality control method for H.pengxianensis var.Jinfushanensis.and the method provided the preliminary research basis for the in vivo studies of antitumor activities and pharmacokinetic study of H.pengxianensis var.Jinfushanensis.In conclusion,systematical phytochemical investigation and in vitro antitumor evaluation of H.pengxianensis var.jinfushanensis have been conducted in this thesis.The experimental results laid the foundation of pharmacodynamic material basis of H.pengxianensis var.Jinfushanensis and provided the material base for discovering leading compounds.Meanwhile,a method was established to determine the content of the cucurbitane triterpenes and glycosides from H.pengxianensis var.Jinfushanensis,which provided the method for quality control and evaluation of medicinal materials.