Receptor Selectivity Of PPARs Agonist HS-602 And Its Pharmacodynamics

Horseradish peroxidase activating receptors(PPARs)are members of the nuclear receptor family,widely exist in body tissues and organs,which associated with many biological activities.There are three subtypes:PPAR??PPAR? and PPARy.Activation of different subtypes can play different pharmacological roles,for example,the lipid-lowering drug Fenofibrate is a PPARa agonist,the lipid-lowering drug GW501516 is a PPAR? agonist,the hypoglycemic Rosiglitazone is a PPARy agonist,the hypoglycemic drug Muraglitazar is a PPARa/y double agonist.Based on obvious differences and closely related between different subtypes of PPARs.Therefore,It not only reduces the side effects caused by single or double agonists,but also has synergistic effect in pharmacodynamics if we develop a PPARs agonist drug that has partial activation of different subtypes,that would have strong development value and market prospect.There is no PPARs three agonist drug has been listed yet,the cytology and pharmacodynamic evaluation system in vivo are also shortcomings such as not comprehensive and systematic.The aim of this study is to construct and improve receptor selection and pharmacodynamic evaluation platform for,PPARs three agonist hypoglycemic agents,analysis of the activation effect of HS-602,an innovative compound developed by Hisun pharmaceutical for the median effective concentration and relative efficiency percentage on activating the different subtypes and pharmacodynamic analysis of various animal models in vivo.Svnthesizing the gene of PPARaLBD-Ga14DB?PPAR?LBD-Ga14DB?PPAR?LBD-Ga14DB,constructing the expression plasmid and report gene plasmid.Transfecting the PPAR??PPAR? and PPAR? plasmid systems into the 293E cells respectively.then incubating with different concentration gradients for 24h respectively.Detecting the luciferase reaction of samples by chemiluminescence method,calculating the relative luciferase activity,drawing the fitting curve,obtaining the EC50 and Max%of activated PPARs subtypes.Selection of positive control samples,deducing the dose required for animal experiment according to its clinical dosage,designing the groups and the number of animals in groups,fast blood glucose?oral glucose tolerance and plasma insulin were selected as evaluation index,then testing the hypoglycemic activity on DIO mice and db/db mice.Another commonly used type 2 diabetic model-ZDF rat was chosen as the research object.Detecting the fast blood glucose?glycosylated hemoglobin and other related indicators of type 2 diabetes,and the blood lipid,liver fat and other related indicators of lipid metabolism.The pharmacodynamics of compound HS-602 in vivo was comprehensively analysed and compared.The receptor activation assay in vitro revealed that the activation effect of compound HS-602 on PPARa was 15%of the positive control GW7647,the activation efficiency of PPAR? was 77%of the positive control GW501516,the activation efficiency of PPARy was 46%of the positive control Piog.The pharmacodynamic in vivo of disease model mice display that the hypoglycemic activity of HS-602 was slightly better than the positive control Rosig,superior to the positive control Piog,and the glucose lowering rate of group HS-602-3 was similar to the Piog-30 group.The level of reducting the plasma insulin was also significantly better than that of the same dose of Rosig and Piog.The pharmacodynamics on ZDF rats f-urther showed that the effect of HS-602 on improving the symptom ms of type 2 diabet and regulating the lipid metabolism disorder is remarkable,and better than the positive control Piog.A method for detecting the affinity and activation efficiency of different subtypes of PPARs was established in this paper,which used for comprehensive evaluation the activation of compounds on different receptor subtypes:Repeated and in-depth studies of compound HS-602 on DIO mice,db/db mice and ZDF rats for Constructing the pharmacodynamic evaluation platform for type 2 diabetes drugs in vivo;A platform for receptor selectivity and in vivo pharmacodynamics of PPARs triple agonists was created groundbreaking which is leading in china,that provides a comprehensive and reliable analysis and evaluation method for the early development of hypoglycemic agents as targets.