Design,Synthesis And Activity Evaluation Of Tyrosine Kinase Inhibitor

Objective:Inrecentyears,malignant tumors are the important diseases threatened the human health,which is becoming the major reason of death.The overall survival of patients with malignant tumor remains poor despite improved diagnostic and treatment strategies.Protein tyrosine kinases(PTKs),a promising therapeutic tumor target,have attracted much attention by virtue of its application in development of antitumor drugs.In our prophase research,compound L029 with high PTK inhibitory activity and good druggability was discovered in a variety of compounds which were reformed from the lead compound SU5416.In this thesis,a series of new compounds with 2-indolone structure were designed and synthesized,with L029 as the lead compound.And their antitumor activity was evaluated by the tyrosine kinase inhibition and cell experiments,then the corresponding structure-activity relationships were analysed and summarized.Methods:1.With L029 as the lead compound,the target compounds were designed and synthesised by preserving the necessary structure of 2-indolone and changing some groups rationally based on the principle of bio-electronic isosteric of molecular design in medicinal chemistry.2.Synthesis of target compounds2.1 Synthesis of type I compounds:The key intermediate(4-Iodo-2-methoxypyridine-3-carboxaldehyde)were prepared from 2-fluoropyridine through Iodine reaction,halogen rearrangement with ethyl formate and sodium methoxide,nucleophilic substitution.Then the compounds(5a~5n)were synthesized from the intermediate through Palladium-catalyzed C-N coupling reaction with aniline derivatives and condensation reaction with 2-indolone.2.2 Synthesis of type II compounds:The compounds(7a~7d)were synthesized from the key intermediate above through metal-free C-O coupling reaction with phenols and condensation reaction with 2-indolone.3.The inhibitory activity of compounds on human colon cancer cells(HCT116),hepatoma cells(HepG2),lung adenocarcinoma cells(H1299),breast cancer cells(MCF-7)and their cytotoxicity on human normal lung fibroblasts(HFL-1)were determined by MTS.The compounds were evaluated through the experiment with five groups(blank,negative,positive/L029,vehicle and compounds group).Their IC50 were measured at five concentrations(100,50,25,12.5,6.25?mol/L)and their survival rate of HFL-1 cells were measured at the concentration of 50?mol/L.4.The inhibitory activity of compounds on PTK were determined by ELISA with five groups(blank,negative,positive/L029,vehicle and compounds group)at the concentration of 100?mol/L and 50?mol/L.Results:1.In this study,18 new compounds 5a~5n,7a~7d were successfully designed and synthesized and their physical and chemical parameters were obtained,their structures were confirmed by ~1H-NMR.2.In vitro anti-tumor activity evaluation results showed that some compounds had good inhibitory activity specifically as follows:5c,7a~7d against HCT116 cells;5c~5e,5i,7a~7d against HepG2 cells,and 5d,5i,7a better than L029;5d,5j,5m,7a~7d against H1299 cells,and 5m better than L029;5b,5d,5g~5k,5m,5n,7a~7d against MCF-7cells,and 5d,5i activity better than L029.3.The cytotoxicity evaluation results showed:5a,5e,7a,7b had good survival rate of HFL-1 cells,and 5a,7a was better than L029.4.The results of tyrosine kinase inhibitory activity showed:5b~5n,7a~7d had certain inhibitory effects on tyrosine kinase at 100?mol/L,however 5a had no inhibitory activity.5b,5d-5g,5i~5n,7a~7c had a certain inhibitory effect,and 5a,5c,5h,and 7d had no inhibitory activity at 50?mol/L.Conclusion:In this paper,18 new compounds were designed and synthesized with L029 as the lead compound through the simple synthetic route with the mild and controllable conditions and convenient post-processing.Some target compounds had good inhibitory activity against tumor cells proliferation and weak cytotoxicity to normal cells,of which 7a was the best.The preliminarily analysed structure-activity relationship of the target compounds revealed that 5-sulfonamido group was a non-essential group possibly,which provided the basis for the further optimized structural design and systematic anti-tumor activity evaluation of this kind of compounds.