Research Of Control Effect And Mechanism To Osteoarthritis By Small-molecule Compounds Targeting FGFR-1

Objective:Since the latest results of researches of gene knockout mouse has been reported,it is indicated that FGFR-1 might be the effective therapeutic target for osteoarthritis.Based on our previous researches,small-molecule compounds,GL-141,targeting FGFR-1,were gained through filtering technique.On the ground of previous cell experiments,we are going to make the assessment of the effect and mechanism of degeneration of articular cartilage and osteoarthritis,to specify the importance of FGFR-1 in the development of osteoarthritis and therapies of targeting FGFR-1,which underlies the next step of the invention of drugs treating osteoarthritis.Method:1.In vivo experiment to clarify the effect of small-molecule compounds to inhibit cartilage degenerationInvestigate the matrix metabolism of cultured chondrocytes extracted from rats.It is demonstrated that the expression of aggrecan is decreased after adding Interleukin 1 alpha(IL-1α).However,adding small-molecule compounds is able to antagonize this kind of effect,inducing the normalization of the expression of aggrecan to the standard groups.2.Assessments of the application of GL-141 on osteoarthritis animal modelsEvaluate the degeneration effect of articular cartilage trauma with the application of small-molecule compounds,GL-141 in multiple trauma-degeneration articular cartilage models of rats and rabbits,and perform pathological analysis of the whole articulation by quantitative score to evaluate the degree of lesion of articular cartilage.3.Elucidation of the cellular molecular mechanism of the effect on articular cartilageInvestigate the effect of small-molecule compounds,GL-141,targeting inhibiting FGFR-1,acting on their downstream signaling pathways including PI3K/AKT,MAPK/Erk and PLC.We also intend to examine the expression of Matrix metallopeptidase 13(MMP-13)and A disintegrin and metalloproteinase with thrombospondin motifs 5(Adamts-5)representing extracellular matrix metabolism,and evaluate cytological changes of articular chondrocytes such as hypertrophy and osteophyte.4.Investigation of the inhibitory effect of the small-molecule compounds to arthrosynovitis in ratsSprague-Dawley rats are injected with collagen to induce arthritis.Small-molecular compounds are injected in articular cavity in the period of induction and the days of 6,11,16,21 after induction.Investigate the difference between the small-molecule compounds group and the saline group pathologically.5.Assessment of security in early stageWith the administration of significant dose(<10uM)through articular cavity and abdomen,we will investigate the aspects of respiratory system and neural system,and clarify whether there are abnormalities in liver,kidney and brain by pathological section.Finally,we can preliminarily assess the pharmaceutical and toxicological effects of these small-molecule compounds.6.Preliminary clinical trial of GL-141 action in human osteoarthritisResults:1,the GL-141 can improve the matrix of animal experimental model of osteoarthritis dissolved,inflammatory cells infiltration and the phenomenon such as cartilage necrosis,can improve the degenerative changes of cartilage of osteoarthritis.2,the GL-141 can inhibit the animal experimental model of FGFR1,MMP-13,Adamts-5,Type II Collagen,Aggrecan and Collagen Type X,the expression of biomarkers of osteoarthritis.3,the GL-141 can relieve the synovial inflammation in the animal model of osteoarthritis,the cartilage cell has certain protective effect.4,the GL-141 role in preliminary clinical trials showed bone arthritis: GL-141 inhibits the degradation of protein,polysaccharide,can reduce collagen type X,MMP-13,Aggrecan and ADAMTS-5 and the expression of p-Akt,,but it does not drop Erk and p-Erk,as well as the expression of Akt.5,high dose application GL-141 was not detected on healthy mice liver,kidney,brain has no obvious pathological effectsThe biomarkers of osteoarthritis such as MMP-13,ADAMTS-5,Type II collagen,Type X Collagen,FGFR-1 are significantly increased in the group of GL-141 compared with control group.The image of these groups indicates the positive effect on the cartilage cells and synovium.Application on mice of high dose of GL-141 didn’t pose a negative effect of animal nodels.Conclusion:1.Applied in animal experiments,GL-141 can mitigate the progress of osteoarthritis,delay cartilage degeneration.2.Cartilage cell lines of cells in the human trials,GL-141 effectively inhibit the expression of osteoarthritis biomarkers,preliminary confirmed that can reduce the progression of osteoarthritis in a certain extent.3.The GL-141 in large dose(<10uM)animal model application no obvious side effects,according to its lower toxicity,safety scope is bigger.GL-141 can effectively inhibit several biological module markers of osteoarthritis.Thus the effectiveness in animal modules of osteoarthritis can be positively confirmed.The toxicities of GL-141 are low and the security ranges of that are wide.