| Among the global issues that affect human health, tumor disease has become second “killer†only to cardiovascular disease. Drug therapy is one of the three treatments for cancer treatment. Although scientists have discovered dozens of effective anticancer drugs, the drug treatment to those most serious solid tumors that endanger human life and health has failed to achieve satisfactory results. Therefore, the development of new anticancer drug screening is still hot.FW-04-806, a bis-oxazolyl macrolide compound with anticancer activity, which was extracted from China-native Streptomyces FIM-04-806, has been reported to be identical in structure to the polyketide conglobatin according to modern organic spectroscopy and x-crystal diffraction experiments. In recent years, the development of antitumor antibiotic macrolide become a hot cancer drug focus, and macrolide antibiotics containing oxazole ring exhibits strong anti-tumor activity, therefore FW-04-806 has a certain research value. Meanwhile, in the preliminary pharmacological and pharmacokinetic pills, FW-04-806 exhibited growth inhibition for multiple tumor cell lines and good pharmacokinetic properties, causing us great concern.Firstly, we carried on the microbiological research on FW-04-806 producing strain Streptomyces FIM-04-806, obtained industrial production strains, exploring fermentation conditions chemical extraction and purification line. We then established testing methods and quality standards of FW-04-806. Secondly, we exploring the pharmacological studies of anti-tumor activity, including anti-tumor spectrum, research targets and its mechanism of action.Specific research methods and results are as follows:1. For Streptomyces FIM-04-806, molecular biology and microbial strain selection were conducted for system bacteria fermentation nutrition and industruial production strains.2. Both shake flask level and automatic pilot fermentor conditions were studied to find out the relationship among fermentation, metabolite changes, strains and production of secondary metabolites conditions. The nutritional supplements and fementation conditions were explored to be suitable for industrition.3. A sound FW-04-806 extration and purification line and quality standards were established, to ensure the quality of FW-04-806 more than 98% purity, single impurity control in less than 0.5 percent.4. MTS assay tested FW-04-806 inhibition of the proliferation of tumor cells. Different concentrations of FW-04-806 treated cells 48 h, and each half inhibition concentration(IC50) were derived according to the result of MTS assay.5. Application of chemproteomics, computational docking, in vitro test between Hsp90 and Cdc37, immunoprecipitation, si RNA gene knock down and Quantitative Real-time PCR on the research of FW-04-806 antitumor mechanism. The result showed that FW-04-806 was a novel Hsp90 inhibitor, directly bound to the N-terminus of Hsp90 and attenuated Hsp90/Cdc37 chaperone/co-chaperone interactions, leading to the degradation of multiple HSP90 client proteins via the proteasome pathway.6. The antagonistic efficacy of FW-04-806 had been investigated at both the molecular and cellular levels. The result showed FW-04-806 inhibited proliferation, caused G2/M cell cycle arrest, induced apoptosis, and downregulated Hsp90 client proteins in a dose and time-dependent manner.7. According to tumor xenografts, FW-04-806 displays a favorable toxicity profile and exhibited potential anti-breast cancer effects in xenograft models.In conclusion, the research carried out FW-04-806 microbial fermentation, purification, quality control and pharmacodynamics pharmacology studies, and the basic medicine evaluation for FW-04-806 had been completed. |
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