The Anticancer Pharmacodynamics Of The T-17,a Dual Target Inhibitor Of CDKs And HDACs

ObjectiveSignaling pathway plays an important role in tumorigenesis.In recent years,many efforts have been focus on the development of molecular target drugs.Combination therapies have been considered as an important strategy due to the complementary anticancer mechanisms.For example,Vorinostat,an approved HDAC inhibitor,have been proved to show synergistic effect with flavopiridol,a CDK inhibitor,in melanoma and neuroblastoma therapy.Studies have confirmed that dual-target drugs are beneficial for solving limited efficiencies such as overcome drug resistance,reduce risks of drug interactions and have predictable pharmacokinetics compared with combination therapies and single target drugs.Herein,our research group have designed and synthesized a novel series of CDKs/HDACs inhibitors with purine as nucleus.The presentative inhibitor T-17 with potent antiproliferative towards A549 cells,HepG2 cells,MDA-MB-231 cells and 4T1 cells,which also showed high potency in vitro HDACs and CDKs inhibitory activity assay.Our study is aimed to evaluate the preclinical bioactivity of inhibitor T-17 and explore the potential anticancer mechanism.In addition,we explored a general synthetic method for the preparation of 1,3,4-oxadiazoles in the design and synthesis of HDACIs.Methods1.The effect of inhibitor T-17 on the proliferation of A549 cells,MDA-MB-231 cells,HepG2 cells and 4T1 cells were measured by CCK8 assay and the effect of inhibitor T-17 on MDA-MB-231 cell migration was evaluated by cell scratch assay.2.Flow cytometry was employed to measure the distribution of cell cycle of MDA-MB-231 and A549 cell intervened by inhibitor T-17.3.Analysis of apoptosis of inhibitor T-17 on MDA-MB-231cells were detected by AO/EB double fluorescence staining,Annexin V/PI staining,ROS staining and Western Blot analysis.4.Western Blot analysis was used to evaluate the expression level of HDACs and CDKs related proteins on MDA-MB-231 cells.5.4T1 xenograft mice models was introduced to evaluated the in vivo antitumor activity of inhibitor T-17.6.The synthesis of 1,3,4-oxadiazole Derivatives Catalyzed by TiCl4was studied.Results1.CCK8 assay and cell scratch assay show that the inhibitor T-17displayed potent antiproliferative activities on four tumor cells.Inhibitor T-17 inhibited proliferation of A549 cells,HepG2 cells,MDA-MB-231 cells and 4T1 cells with IC₅₀ values of 3.75±0.54μM,3.58±0.63μM,2.41±0.95μM and 10.67±0.67μM,respectively.2.Inhibitor T-17 arrested MDA-MB-231 and A549 cells cycle at G2phase and S phase,respectively.3.Inhibitor T-17 induced the apoptosis of MDA-MB-231 cells.4.Western Blot assay displayed that inhibitor T-17 can significant down-regulate the expression of p-CDK2,CDK2,Cyclin E and up-regulate the expression of AcH3.5.Inhibitor T-17 showed good anti-tumor activity in vivo studies.6.We found a new one-pot synthetic method of 1,3,4-oxadiazole Derivatives.A total of 15 derivatives were synthesized,one of which was new.ConclusionThis study showed that inhibitor T-17 possessed potent antitumor activity against four tumor cells in vitro and in vivo models.We explored the antitumor mechanism of inhibitor T-17 using Western Blot analysis,which indicated that HDACs and CDKs signaling pathways were involved in the anticancer activity of inhibitor T-17.These results laid a foundation for the further study of inhibitor T-17.In this study,the synthesis route of 1,3,4-oxadiazole has been studied and a new one pot synthesis route for 1,3,4-oxadiazole by means of TiCl₄ catalysis has been developed,which would be beneficial for the synthesis of HDAC inhibitors or dual CDKs/HDACs inhibitors.