The Anti-Cancer Activities Of1122-37, A Novel Derivative Of Sorafenib, And Its Mechanism

Backgrounds:Receptor Tyrosine Kinase inhibitors (RTKi) are a kind of molecular targeted small molecule for chemotherapy. The advance of RTKi changed the traditional non-targeted way of chemotherapy, gained better curative effect than that of traditional chemotherapy drugs. Sorafenib, which is broadly used in clinical chemotherapy of hepatocellular carcinoma (HCC), renal cell carcinoma (RCC) and non-small cell lung carcinoma (NSCLC), is one superior representative of RTKi. However, the side-effects of sorafenib restrict its usage, including hepatotoxicity, renal toxicity, and cardiotoxicity. Therefore, it is a research focus that using the structural remould based on the sorafenib structure to find new chemicals with high effect.1122-37is a indazole compound based diarylurea derivatives which possesses better activity than sorafenib according to the preliminary tests. In this research, we estimated the anti-cancer activity of1122-37in several ways in vitro and explored its mechanism.Methods:The inhibitory effects of1122-37on PLC/PRF/5cells proliferation was estimated by the3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) method. Hoechst33258staining and Annexin V/PI staining assays were utilized to determine the apoptosis induced by1122-37on PLC/PRF/5. PI staining was performed to evaluate the cell cycle arrest of PLC/PRF/5induced by1122-37. Western blotting assay was used to analyze the related proteins for deducing the mechanism of1122-37. The capillary tube formation assay was performed to evaluate the activity of1122-37against capillary tube formation of HUVECs on3-D Matrigel, and its mechanism was evaluated by western blotting assay, too.Results:The anti-proliferation effect of1122-37was estimated by MTT assay. Statistical analysis indicated that the IC50value that based on the inhibition rates of72h exposure was4.34μM. The apoptosis inducing effect of1122-37was estimated by Hoechst33258staining, FITC-Annexin V/PI staining, and western blotting.1122-37(1.25-5μM) induced the hyperchromatic nuclear, chromatin gathering around to the nuclear membrane, or splintering in PLC/PRF/5cells after24h treatment, the apoptosis ratio of PLC/PRF/5cells were16.08%,24.73%and29.26%after exposed24h of1.25,2.5and5μM1122-37,1122-37might induce PLC/PRF/5apoptosis through reducing the level of Mcl-1and triggering the caspase cascade. The cell cycle arresting effect of1122-37was estimated PI staining and western blotting. The ratio of PLC/PRF/5cells which stayed in G1phase were78.67%,80.98%and83.03%after exposed24h of1.25,2.5and5μM1122-3711122-37might induce the cell cycle arrest of PLC/PRF/5through reducing the level of cyclin D1. Further analysis of western blotting suggested that these effects of1122-37might arise from its roles in the inhibition of multi-kinases, including c-Kit and its downstream targets and the Wnt/β-catenin pathway in PLC/PRF/5cells. The antiangiogenesis effect of1122-37on HUVECs was estimated by MTT assay, capillary tube formation and western blotting.1122-37(5-25μM) indicated the growth of HUVEC and the the capillary tube formation, might arising from its effect of inhibition of VEGF and phosphorylation of VEGFR-2.Conclusion:1122-37possessed the activity against PLC/PRF/5cell proliferation. We suggested that these high effects of1122-37might arise from its roles in the apoptotic induction and regulation of cell cycle. The mechanisms of1122-37action might associate with its activity in the inhibition of multi-kinases.1122-37strongly prevented HUVECs capillary tube formation. The effect of antiangiogenesis might be due to the prevention of VEGF and phospho-VEGFR in HUVEC. The activity of 1122-37against PLC/PRF/5proliferation and HUVEC angiogenesis were greater than sorafenib. We suggested that1122-37might be a promising compound that could be developed as a potential agent for supplant the use of sorafenib.