Design,Synthesis,antitumor Activity And Mechanism Of Chrysin Derivatives

The incidence of tumor is increasing day by day,and the treatment of tumor has become a hot research topic in medical and other related industries.Natural products have become hot spots in the research of many anti-tumor drugs.For example,natural products such as paclitaxel and camptothecin have been proved to have good anti-tumor activity.Therefore,the discovery of new natural products with anti-tumor activity or the synthesis of target compounds with natural product structures have attracted extensive attention,and the exploration of new anti-tumor mechanisms of natural products or small molecules of drugs has become increasingly important.In this study,we reasonably spliced drug skeletons on chrysin skeletons,synthesized chrysin derivatives and explored their anti-tumor effects.In order to obtain more effective and less toxic anti-tumor lead compounds and provide theoretical basis for the research and development of chrysin derivatives anti-tumor drugs.（1）The synthesis of chrysin derivatives.Exploring at the early stage of the team,on the basis of chrysin as starting material,in calcium hydroxide alkaline methanol solvent and intermediate compounds of each replaced（2-toluenesulfonic malononitrile）cyclic addition reaction,eventually get skeleton of a natural chrysin derivative 3a to 3r,a total of 18,its production rate is77-91%of all the synthesized compounds structure by ¹H NMR,¹³C NMR were identified by MS-ESI,and obtained by X-ray single crystal diffraction technology of compound 3a crystal configurations.（2）The 18 compounds were screened for anti-tumor activity in vitro.K562、PC-3、A549、NCI-H1299 four cancer cells were selected for in vitro antitumor screening by MTT assay（tetramethylazolate microenzyme reaction colorimetry）.The reaserch showed that the compound 3h had the strongest antitumor activity against K562 cells,with an IC₅₀ value of 6.41μM.Compound 3b showed the strongest antitumor activity against PC-3 cells,with an IC₅₀ value of 26.08μM compound 3b showed the same antitumor activity against A549 cells as the positive control drug cisplatin,and its IC₅₀ value of 15.39μM compound 3g showed the same inhibitory activity against NCI-H1299 cells(IC₅₀=18.76μM)as the positive control drug cisplatin(IC₅₀=21.74μM).（3）Study on the mechanism of the inhibition of K562 cell proliferation by compound 3h.Morphological changes,AO/EB fluorescence staining and Hoechst33258fluorescence staining of K562 cells treated with compound 3h were investigated by inverted microscope.The results showed that K562 cells showed different degrees of apoptotic statology after 3h treatment.Flow cytometry was used to detect the apoptosis level of K562 cells,indicating that the percentage of apoptosis of K562cells after 3h treatment increased gradually with the increase of treatment concentration or the extension of action time,and the cells were more inclined to late apoptosis.Flow cytometry analysis of K562 cell cycle showed that the compound was inhibited in G1/S phase 24 h after 3h treatment of K562 cells.Western blot showed that the protein expressions of p53 and Bax in K562 cells were significantly up-regulated and the protein expression of Bcl-2 was down-regulated at the same time.（4）Study on the mechanism of compound 3b inhibiting the proliferation of A549cells.The inhibitory effect of alicin derivative 3b on A549 cells was investigated from the aspects of cell morphology,soft AGAR colony,cell scratch,Transwell invasion and migration,and apoptosis staining analysis.The results showed that compound 3b could inhibit the proliferation of A549 cells and induce the apoptosis of A549 cells,and the degree of apoptosis was concentration and time dependent.（5）Study on anti-tumor mechanism of compound 3b in vivoThe Lewis lung cancer model of C57BL/6 mice was established,and the tumor formation rate of each group was 100%.The results showed that the tumor growth was significantly inhibited after the treatment of compound 3b,which showed a certain dose dependence,indicating that compound 3b had a good anti-tumor effect in vivo.In conclusion,this thesis by chrysin to nuclear structure,reasonable design and synthesis of a series of chrysin derivative with antitumor activity in vitro screening,get chrysin derivative 3h showed better inhibition of proliferation activity to K562cells,3b on A549 cell phenotype out better inhibition of proliferation activity,and to explore ways of role,preliminary reveals the compound 3h、3b antitumor mechanism;Through the study on the anti-tumor mechanism in vivo,compound 3b has a good anti-tumor effect in vivo,which provides a theoretical basis for the research and development of anti-tumor drugs of alicin compounds,and has certain theoretical value and guiding significance.