Study On The Efficacies And Mechanisms Of Compound B192and EGCG

Cancer is becoming a serious human healthy problem. The incidence of cancer is increasing in world-wide as a result of the change of the environment, as well as increasingly, an adoption of cancer-associated lifestyle choices. There are still many problem of the therapy of tumor, because of the limited knowledge of the development of different of kinds of cancer. In addition, the prevention of the happening of cancer is becoming more and more important for the therapy. Becuae of the difficult of the cure of cancer, it is a meaningful way to develop the therapy of the precancerous disease. The precancerous disease mainly refers to the disease that has a high conversion rate to cancer. If we find a way to inhibit the occur of precancerous disease, the incidence of cancer will decrease significantly. Therefore, screening and identification of new therapeutic agents for precancerous disease are also important. In summary, our study is based on two aspects:the screening of new antiancer small molecule drug and the tactics of prevention of cancer.At present, the research and development of novel small-molecular targeted anti-cancer drug is mainly based on the crystal structures of targeted proteins which are closely related to tumor occurrence, development and maintenance and therational drug design by computer aided design technology. And finally discover the targeted, low toxicity and effective lead compound through oriented synthesis, SAR studies, furtherstructural modification and systemic pharmacological assessment methods. So the study of small-molecular targeted anticancer drugs is one of the most important research frontiers in drug discovery. In the first part of our study, according to the method of scaffold hopping, we designed and synthesized the3-amino-1,4,5,6-tetrahydropyrrolo3,4-c]pyrazole derivatives B192which was based on two small molecule inhibitors in clinical trials. The result showed that B192greatly inhibited cell proliferation in various type of human cancer cells. In the kinase inhibition assay, B192had slight inhibiton effect on CDK5/p25and GSK3β, and little inhibiton effect on the CDK2kinase. The cytotoxicity of B192for human normal liver L02cells was smaller than cancer cell lines. According to the growth curve and colony formation assays, B192inhibited the proliferation of colon cancer cells HCT116in a time-and dose-dependent manner. The flow cytometric analysis revealed that B192induced cell cycle arrest at G2/M phase and resulted in a significant increase of HCT-116cell apoptosis in a time-and dose-dependent manner. It also caused cell cycle arrest at G2/M in HCT116p53-/-and HT29cells. The results of Western Blot showed that B192inhibited the expressions of cdc25C,cdc2,Cylin B1. the expression of p53was not change in HCT116cell and HCT116p53-/-cells. The HCT116p53-/-cell did not express p53, but the levels of p21was significantly increased. The expression of p-Akt, p-ERK1/2and NF-κB were decreased after treated by B192for12h. In vivo experiment showed that B192slightly inhibited the growth of CT26cells by24.13%in tumor weight.Becuse of the solubility of B192, it didn’t have a high inhibition effect on the tumor growth in vivo. B192still had a good inhition effect in vitro, it is also provided a theoretical basis for deveolpment a new target small molcecule drugs.Liver fibrogenesis is common to various types of chronic liver disease.The possible consequences of fibrogenesis include liver cirrhosis or hepatocelluar carcinoma.(-)-epigallocatechin-3-gallate (EGCG) is one of the most abundant polyphenols present in green tea. EGCG is well known for its anti-tumor activity while other pharmaceutical effects were discovered subsequently. It is reported that EGCG had effect on the liver fibrosis on the model induced by CCl4. Currently, our therapeutic repertoire for the treatment of liver fibrosis and cirrhosis is severly limited. To study the protective effects of (-)-epigallocatechin-3-gallate (EGCG) on hepatic fibrosis induced by bile duct-ligated rats and their mechanisms. The result of our study showed that EGCG significantly reduced the expression of fibrotic marker genes and protein in a human HSC line and inhibited the fibrotic marker protein expression via PI3K/Akt and TGF-β/Smad signaling pathways in LX-2cell line. EGCG reduced the liver fibrosis in BDL rat via blocking PI3K/Akt and TGF-β/Smad signaling pathways. Knock down G3BP1/2and transient high expression G3BP1/2didn’t influence the expression of fibrosis-related genes. It is suggested that G3BP was not a new antifibrotic target and the antifibrotic effect of EGCG was not by inhibition of G3BP. In conclusion, we found a new mechanism of the antifibrotic effect of EGCG, and these novel findings suggest that EGCG may be a new drug for the treatment of liver fibrosis.Our research investigated the anticancer effect of B192, one novel compound designed as a small targeted anticancer drug and discussed the mechanism of the anticancer effect. Though B192has poor water solubility, which is the major limit for ability of becoming a new drug, our work contributed to further design and synthesize better targeted and has better solubility compounds. One the other hand, we focused on the cancer prevention. We confirmed the antifibrotic effect of EGCG on BDL model and discovered new insights in the mechanism of antifibrotic effect.In summary, our research demonstrated the mechanism of novel compound B192, providing support for further design, synthesis and investigation of small molecule drugs. Our research also revealed that novel antifibrotic mechanism of EGCG, which contributed to the investigation of antifibrotic drugs. Taken together, our research focused on the anticancer small molecule compound and antifibrotic compound EGCG,supporting for further cancer therapy and cancer prevention.