Design, Synthesis And Evaluate Of Bcl-2 Protein Inhibitors

Cancer is one of the most common and frequently- occurring diseases which is now threatening the people's health and life seriously. According to WHO statistics, in 2004 the number of cancer deaths is 7,400,000(about 13% of all deaths), more than 70% of cancer deaths occurred in low and middle income countries. It is expected that the cancer mortality rate will continue increasing, the number of cancer deaths will be 120,000,000 till 2030.Currently, the main anti-cancer drugs are cytotoxic drugs in clinic. It is difficult to avoid some disadvantages such as low selectivity, toxicity, and easy to produce drug resistance in using this type of anti-cancer drugs. In recent years, with the rapid progress in life science research, the basic process such as the signal transduction, regulation of cell cycle, and apoptosis in the malignant cells is being gradually clarified. Some protein which related to apoptosis has been recognized as the target of drug design.Bcl-2 protein is one of the key factors controlling apoptosis which get close affinity to the occurrence of tumor. And the tumor cell just over-expression Bcl-2 protein, so it has been widely recognized as a potential anti-cancer drug target. TMP20 is a novel and effective inhibitor of Bcl-2 protein. It was designed through computer-aided visual screening and modification. From the BIOCORE-SPR (Biological Collaborat -ive Research Environment and Surface Plasmon Resonance) and NMR tests we claim that TMP20 binds to the Bcl-2 protein effectively. Cell experiment showed that TMP20 is highly selective in inhibiting the growth of Hela cell which express Bcl-2 protein in high level. Animal test also showed that TMP20 is efficient in inhibiting tumor growth in a xenograft mouse model.In this study, we chose TMP20 as lead compound and design some new derivatives according to the known structure-activity relationship. We have synthesized 24 new compounds. All compounds were characterized by MS and 1H-NMR. According to the results of activity evaluation, 8 compounds showed potential activity such as LK-A013, LK- B023, LK-A036, LK-B030, LK-B044, LK-B047, LK-C05, LK-B039, and LK-B038. Further biological evaluation is still in progress.Objective: Acquire some more effective Bcl-2 protein inhibitors.Methods: Since 1998, Institute of Pharmacology & Toxicology, Academy of Military Medical Sciences has researched in the Bcl-2 protein inhibitors as anti-tumor drugs. They have synthesized more than 200 compounds, and some of them have novel activities such as Z24, TMP20. In this study, according to the known structure-activity relationship, we designed three types of target compounds, I, II and III type, a total of 24 target compounds.According the relevant literature and the experiment, we find the synthesis route, and the optimum reaction conditions of all reactions such as solvent, temperature, and the reaction time. Then we synthesizes the target compounds according the routes and conditions, acquired 4 target compounds of I type via 6 steps; 2 target compounds of II type via 9 steps; 18 target compounds of III type via 11steps.We test the activities of the target compound by MTT method on the HL60 cell and the IM9 cell.Results: 1 we synthesized 12 key intermediate, some of structure was identified by 1HNMR and melting point.2 We synthesized 24 target compounds, which structure was identified by 1H-NMR, MS. Some of them was identified by 13C-NMR, H-H COSY, NOE.3 According the activity tests, we find LK-A013, LK-B023, LK-A036, LK-B030, LK-B044, LK-B047, LK-C005, LK-B039 have potential activity (Table 1).Conclusion: All of the 24 target compounds showed certain inhibition activities, 8 compounds showed novel activities. So the design of our study was rational, after further research, some of the novel compounds will be the new Bcl-2 inhibitors.