Design,Synthesis And Biological Evaluation Of Chromeno[4,3-c] Pyrazol-4(2H)-one Containing Carbonyl Or Oxime Derivatives As Potential,Selective Inhibitors PI3K?

Malignant tumor possesses the ability of invading the surrounding tissues and transferring to the distant tissues.if its grow and spread can not be controlled,it will damage the function of normal viscera and finally lead to death.Therefore,cancer is serious endangering human' s health.Especially in recent years,the incidence and mortality of tumor diseases continuous rises,which indicates cancer is becoming the human health "killer".The traditional small molecular drugs has a long history,but it has high toxicity and less selectivity.PI3K/AKT/mTOR can control a series of important cellular activities including proliferation,survival,growth,apoptosis and so on.At the same time,a large number of studies have shown that the process of tumor growth is commonly associated with the high expression from PI3K in the cell.Tumor cells complete the cell proliferation through activating the cascade reaction of PI3K.Consequently,taking PI3K as a new target to design anti-cancer drugs becomes the important hot spot in the research of treating cancer.In this research,we focused on the design,synthesis and biology evaluation of novel molecule as potent PI3Ka inhibitors.Several natural products as PI3K inhibitors are widely known,including first-generation PI3K inhibitors such as wortmannin,LY294002 and its analogues.It was a feasible program that chromeno[4,3-c]pyrazol-4(2H)-one was designed as the core region.On one hand,we introduced acetophenone fragment into the derivatives in order to they can well-bonded with the PI3K protein.On the other hand,carbonyl of acetophenone was then replaced by oxime,for this modification might enhance solubility,kinase inhibition and cell permeability.We have used the Ligand Fit Dock protocol of Discovery Studio to screen quantities of designed compounds,and a series of novel chromeno[4,3-c]pyrazol-4(2H)-one containing carbonyl or oxime derivatives(4a-4n,5a-5n)have been synthesized.All of the synthesized compounds gave satisfactory analytical and spectroscopic data in full accordance with their depicted structures.Additionally,the structure of compound 4a and compound 5b was further confirmed by X-ray diffraction.Fig.1.3D model of the interaction between compound 51 and 3HHM binding site of PI3Ka proteinAs for biological activity,the results of antiproliferative against cancer cells showed that most of the synthetic compounds displayed remarkable antiproliferative effects on the cell lines,and the compounds(5a-5n)that contain the oxime showed a remarkable improvement versus its counterpart(4a-4n)containing the carbonyl in the inhibition of HCT-116 cell proliferation.Out of them,compound 51 showed the most potent antiproliferative activities against HCT-116 with IC50 of 0.10 ?M in vitro,which was superior to the positive control LY294002.Compound 51 exhibited most efficient inhibitory activity with IC50 of 0.012?M.A further study comparing the antiproliferative activity against the HCT-116 cell line with the PI3Ka inhibitory activity and the result revealed that the potent anticancer activities of synthesized compounds were likely related to their PI3K? inhibitory activities.In addition,the results of apoptosis study show that 51 induced apoptosis in a dose-dependent manner.Docking simulation of 51 into PI3Ka active site were performed to determine the probable binding model,and there are three amino acids VAL851,LYS802 and ILE848 located in the binding pocket playing vital roles in the conformation with compound 51(Fig.1).All above,the molecular docking result along with the antiproliferative activity date,suggested that compound 51 is a potential inhibitor of PI3Ka.