Study On The Chemical Constituents And Biological Activity Of Zanthoxylum Dimorphophyllum Hemsl From Guizhou Province

Objective: The purpose of this research is to systematically study the chemical components of Zanthoxylum dimorphophyllum Hemsl from Guizhou province.The pharmacological activities of Z.dimorphophyllum were systematically screened according to the traditional folk medicinal efficacy,and the natural lead compounds with medicinal value were found to provide theoretical basis for the deep development and utilization of Zanthoxylum heterophyllum resources.Method: The leaves and twigs of Z.dimorphophyllum was dried and crushed,and extracted under refluxing with 75% methanol.The methanolic extract of Z.dimorphophyllum were successively fractionated by silica gel column chromatography,MCI column chromatography,Sephadex LH-20 column chromatography and further purified by HPLC to obtained the compounds.The structures of the compounds were identified by NMR,MS,IR,UV and specific rotation,the absolute configuration of the chiral compound was determined by calculating ECD.Screening compounds for neuroprotective activity against corticosterone damage(PC-12 cells).Evaluation of antitumor activity of compounds using MTT method(A549 and MCF-7 cells),determination of tyrosinase inhibitory activity using dopa oxidation assay(B16-F10cells).In addition,DPPH and ABTS methods were used to screen the antioxidant activity of the compounds.Result: A total of twenty-one compounds were isolated and identified from the ethyl acetate fractions of Z.dimorphophyllum from Guizhou Province,including ten coumarins(1-10),one alkaloid(11),three lignans(12-14)and one flavone(15),six phenolic acids(16-21).Compounds 2,3,5,7-10,12 and 16-21 were isolated from Z.dimorphophyllum for the first time,compounds 14 and 15 were first isolated from Rutaceae.The protective effects of compounds 12,13,19,and 21 on PC-12 cells damaged by corticosterone showed significant protective effects at concentrations of20 μM.Antitumor activity showed that compounds 4,14 and 18 had certain inhibitory activity on the proliferation of A549 lung cancer cells and MCF-7 breast cancer cells at a concentration of 100 μM.The tyrosinase inhibitory activity showed that at a concentration of 100 μM,the inhibitory rates of the selected compounds on tyrosinase were all lower than 50%,and the selected compounds did not significantly inhibit the activity of tyrosinase.The results of antioxidant activity screening showed that the DPPH method showed that six compounds had significant DPPH free radical scavenging ability,and the activities of compounds 13,14 and 18 were higher than those of positive control L-ascorbic acid;ABTS method showed that nine compounds had significant antioxidant activity,among which compounds 7,8,9,12,13,14,18,and 19 had higher antioxidant activity than the positive drug VE.Conclusion: In this study,twenty-one compounds were isolated and identified from the ethyl acetate fraction of Z.dimorphophyllum,the main chemical components in Z.dimorphophyllum are coumarins,alkaloids,lignans.The effects of these compounds on anti-tumor,tyrosinase inhibition activity and protection of PC-12 nerve cells were screened,which provided important material and theoretical basis for the follow-up development and utilization of Z.dimorphophyllum.