Preliminary Study Of The Pharmacological Properties Of Methylene Modification Derivatives Of Dapagliflozin

Objective:SGLT2 inhibitor is a novel hypoglycemic drugs,SGLT-2 inhibitor can increase the urinary glucose excretion,then reduce plasma glucose levels.Dapagliflozin is the first to market.Structural modification in existing drugs,in order to find new and effective drugs has been an important part of drug development.Modification between the benzene ring methylene,systematic evaluation the derivatives in efficacy,pharmacokinetics and toxicity in order to find the regularity,provide the basis for the further transformation.Methods:To evaluat the inhibitory effect of the drug on the cellular level in vitro and obtain the suppressing IC50 values in SGLT1 and SGLT2 of the compounds,calculated selectivity to the SGLT1 and SGLT2.Through the rats in vivo tests,obtained the urine output,the urinary glucose excretion and oral glucose tolerance test results.Long-term experiments conductedthe effects of the compounds in STZ-induced diabetic rats,measured the fasting plasma glucose and fructosamine.Measured the pharmacokinetic parameters in oral and intravenous pharmacokinetic experiments in rats,preliminary find out the absorption properties of the compound.Acute toxicity of drugs in mice and rats long-term toxicity studies,preliminary find out the changes in toxicity of the compounds.Results:1.In vitro cell experiments,the SGLT2 IC50 value of Dapagliflozin was 0.0012 μM,the SGLT2 IC50 value of one methyl-substituted product(TY702-5)was 0.0337 μM,the SGLT2 IC50 value of dimethyl-substituted product was 0.2529 μM.After transformation the SGLT2 inhibition of the derivative are decreased.The selectivity of SGLT2,one methyl product decreased 5 times,dimethyl-substituted compound decreased about 30 times.2.Urinary glucose excretion and oral glucose tolerance test showed that,compared with the model group,during 0-6 h and 6-24 h urinary glucose excretion and 0-3 h glucose inhibition rate,all the compounds showed a dose-dependent increase(P<0.05);under the same dose,TY702-5 and TY702-5D during 0-6 h and 6-24 h urinary discharge sugar and 0-3 h glucose inhibition rate was significantly lower than that Dapagliflozin group(P<0.05);3.Long-term administration experiment in STZ-induced diabetic rats showed,the administration could significantly reduce fasting plasma glucose and fructosamine in a dose-dependent manner.High-dose group 90 mg/kg were significantly different with the model group.4.In vivo absorption test,compared with Dapagliflozin,one methyl-substituted product(TY702-5)showed the absorption rate(Tmax)and extent(Cmax and AUCo-t)increased,elimination half-life(T1/2)substantially constant,tissue distribution(Vd)and bioavailability(F)decreases.Dimethyl-substituted product(TY702-5D)showed,the drug absorption rate(Tmax)accelerateed,the extent of absorption(Cmax,and AUC0-1)decreased,tissue distribution(Vd)increased,elimination half-life(T1/2)and bioavailability(F)decreased.5.The results of acute toxicity in mice showed that,the mortality rate of high-dose group is higher than the low-dose group,in a dose-dependent manner,Dapagliflozin 450 mg/kg groups of female mice died 3,TY702-5D 450 mg/kg groups of female mice died 1,TY702-5 450 mg/kg group of male mice died 1.STZ-induced diabetic rats showed that long-term administration had dose-dependent toxicity,the 500 mg/kg group of Dapagliflozin and TY702-5D,the female rats all died,male group were three and two deaths respectively,TY702-5 death only one female rats in high-dose group.Dapagliflozin and TY702-5D mainly have the thymus,spleen,adrenal glands and brain toxicity,while significantly affect hematological and biochemical indicators.TY702-5 had low toxicity and less impact on the rats.Conclusion:Methylene position between the two benzene rings in dapagliflozin is a great influence position.The volum of substituents had a negative correlation with activity,particularly,the volume of substituents had tremendous impact on absorption,blood exposure and elimination rate,suggesting the modification of this position requires a comprehensive and balanced strategy,the modification in this position need a adjusted in other parts of the structure.