Based On P-gp,Bcrp,and Mrp2 Exploring The Effects Of Telmisartan On The Pharmacokinetics Of Canagliflozin

Part one Effects of telmisartan on the Pharmacokinetics of canagliflozin in ratsObjective:To develop a method for the detection of canagliflozin in rat,and to evaluate the Effects of single and multiple administration of telmisartan on the Pharmacokinetics of canagliflozin.Methods:Twenty four male rats were randomly separated into four groups:Group A and B animals were administered canagliflozin with and without telmisartan for a single day,respectively.Animals in group C and D were administered canagliflozin with and without telmisartan for seven consecutive days,respectively.The administered doses were 8 mg/kg for telmisartan and 10 mg/kg for canagliflozin.Plasma was collected according to designed time points,and the rat plasma samples were analysed by UPLC-MS/MS after liquid-liquid extraction,and the chromatographic separation was performed on a Waters BEH C18 column,and the mobile phase is 0.1%formic acid water and acetonitrile,gradient elution,positive ion detection.The non-compartmental model was used to calculate the Pharmacokinetics parameters of canagliflozin by DAS 2.1 software.Results:The calibraton curve for canagliflozin in rat plasma have a good linearity,ranging from 5-3000 ng/m L,the plasma methodologies met the requirements.The AUC₀–_48h and AUC₀–∞of canagliflozin in the combined treatment group were significantly increased by 57.7%and 56.6%,respectively.In contrast,after a seven-day administration,the AUC_0-48h and AUC_0-∞of canagliflozin significantly decreased by 38.0%and 36.1%,respectively.Conclusions:Telmisartan increased the AUC_0-48h,AUC_0-∞of canagliflozin in rat plasma after a single-day administration,but decreased the AUC_0-48h,AUC_0-∞of canagliflozin in rat plasma after a seven-day administration regimen.Part two Effects of telmisartan on the tissue distribution of canagliflozin in miceObjective:To develop a method for the detection of canagliflozin in tissue（liver homogenate samples as represent）,and to evaluate the Effects of telmisartan on the tissue distribution of canagliflozin after seven-day administration.Methods:Forty two male mice were randomly separated into two groups:group A and B were administered canagliflozin with and without telmisartan for seven consecutive days,respectively.The administered doses were 12mg/kg for telmisartan and 15 mg/kg for canagliflozin.Tissue was collected according to designed time points,and the tissue homogenates were analysed after liquid-liquid extraction.The detection condition is the same as that of plasma samples,the non-compartmental model was used to calculate the Pharmacokinetic parameters of canagliflozin by DAS 2.1 software.Results:The calibraton curve for canagliflozin in tissue homogenate samples have a good linearity,ranging from 5-3000 ng/m L,the liver homogenate samples methodologies met the requirements.Canagliflozin was widely distributed in all collected tissues.After a seven-day administration,the highest concentrations of canagliflozin were detected in the kidneys,followed by the intestine,liver,lung,heart,spleen,and brain,for both groups.In the canagliflozin group,the concentration of canagliflozin showed an obvious double-peak phenomenon,at approximately 4 h and 8 h.However,in the combined treatment group,the concentration of canagliflozin showed a peak at 2 h.Furthermore,the concentrations of canagliflozin in the kidney liver,lung,and heart at 2 h were significantly higher in the combined treatment group compared to the group treated with canagliflozin alone.Conclusions:After a seven-day administration,the highest concentrations of canagliflozin were detected in the kidneys,followed by the intestine,liver,lung,heart,spleen,and brain,for both groups.Furthermore,the concentrations of canagliflozin in the kidney,liver,lung,and heart tissue at2 h were significantly higher in the combined treatment group compared to the group treated with canagliflozin alone.Part three Effects of telmisartan on efflux transport proteins expression in mice liver and kidneyObjective:To investigate the Effects of telmisartan on the protein expression of P-gp,Bcrp and Mrp2 in liver and renal.After seven-day dosing,the Pharmacokinetic mechanism of canagliflozin was explored from the perspective of the efflux transporters.Methods:Based on the results of tissue distribution,after canagliflozin administering for 2 h,the expression of efflux transporter proteins in the liver and kidney were evaluated by Western blot.The data was analysed by Image J1.8.0 and SPSS 25.0 software.Results:Compared with the group treated with canagliflozin alone,the expression of P-gp protein was similar,the expression of Bcrp protein increased by 44.6%,and the protein expression of Mrp2 significantly increased by 64.9%in mice liver;as for kidney,the expression of P-gp protein significantly increased by 79.7%,and the expression of Bcrp and Mrp2 protein increased by 35.6%and 14.7%,respectively.Conclusions:Telmisartan may compete with hepatic Mrp2 and renal P-gp,increasing the expression of efflux transporter proteins,thereby increasing the concentration of canagliflozin in tissue.