The Isolation And Identification Of Antitumor-active Phthalides Targeting Nuclear Receptor TR3 From Angelica Sinensis

Recent years,molecular targeted drugs have become a new trend of research and development of new anti-cancer drugs.As one of the targets,nuclear receptors are especially closely watched.TR3/Bcl-2 apoptosis pathway is an unique and effective regulation pathways of cancer cells apoptosis with the orphan nuclear receptor TR3 participation.The aim of our study was to isolate the compounds targetedly from the Chinese herb Angelica sinensis,and screen out compounds targeted nuclear receptors TR3 with potential anticancer function.In this study,28 kinds of compounds were obtained from Angelica sinensis by chromatographic separation methods.Their chemical structures were identified by physicochemical properties and modern wave analysis methods,including ten monomeric phthalides:Z-Ligustilide(1),E-Ligustilide(2),Butyl phthalide(3),Senkyunolide G(8),Z-8-hydroxy-butylidenephthalide(15),Z-6,7-epoxyligustilide(18),Senkyunolide E(24),5-hydroxy-3-butylphthalide(25),Senkyunolide C(26),Senkyunolide B(27),eleven dimer phthalides:Angeolide(6),Angeolide methyl ester(7),Z,Z’-6’.6,7.3’a-diligustilide(9),Z-6.8’,7.3’-diligustilide(10),3Z’-3S,8R,3a’S’,6’R-4,5-dehydro-3.3a’,8.6’-diligustilide(11),3.3a’,8.6’-diligustilide(12),Z-8’.3a,3’.6-diligustilide(13),3Z,3Z’-6R*,7a’S*,7S*,3a’S*-6.7a’,7.3a’-diligustilide(19),Senkyunolide O(23),3Z,3’Z-6R*,3a’S*,7R*,7a’R*-6.3a’,7.7a’-diligustilide(20),3’Z-3 R 8S 3a’R,6’S-3.3a’,8.6’-dligustilide(28),seven other compounds:3R,8S-Falcarindiol(4),Coniferylferulate(5),E-Coniferyl Ethyl Ether(14),Ferulic acid(16),Trilinolein(17),9,12-Octadecadienoic acid(Z,Z)-2,3-dihydroxypropyl ester(21),Dibutyl phthalate(22),among them,19 and 20 were novel compounds.In this study,the results showed that the 12,14,20 could significantly activate TR3 by means of the nuclear TR3 reporter gene screening model.At the level of protein expression,western blotting showed that 2,24,12 have signifiicant effect on TR3 protein expression upregutation.The MTT was used to screen anti-tumor effect of cells,and 12,28 with 50 μM were cytotoxic to human lung cancer cell line H460.And 12 can be researched further as a potential anti-tumor compound through TR3/Bcl-2 apoptosis pathway.