Hepatic Transporters-based Study On The Effects Of Hesperidin On Pharmacokinetics And Intestinal Toxicity Of Irinotecan And Active Metabolite SN-38

Aims: To investigate the effects of hesperidin on the pharmacokinetics and intestinal toxicity of irinotecan(CPT-11)and its metabolite SN-38,and explore the underlying mechanism based on hepatic transporters.Methods: 1.Developed an exclusive,rapid,sensitive and reliable method for determination of CPT-11 and SN-38 in biological samples using LC-MS/MS.2.Investigated the effects of normal-dose of hesperidin(Hesd-20,20 mg/kg)or high-dose of hesperidin(Hesd-200,200 mg/kg)single administration on the pharmacokinetic profile,tissue distribution,and biliary excretion of CPT-11 and SN-38 in rats.3.Investigated the effects of multiple dose of hesperidin(Hesd-20,20 mg/kg × 2 d)on pharmacokinetic profile,tissue distribution,biliary excretion and intestinal contents of CPT-11 and SN-38 in rats.4.The protein expression of related hepatic transporters were evaluated by western blot technique and immunohistochemistry.5.Investigated the effects of hesperidin on intestinal toxicity of CPT-11 induced delayed-onset diarrhea(CIDD)model by evaluating body weight,diarrhea score,and colonic pathological alteration in rats.Results: 1.The linear range of CPT-11 and SN-38 were 2-20000 ng/ml and 2-10000 ng/ml,respectively;and the specificity,precision,stability,recovery rate,and matrix effects were in conformity with the provisions in the developed method.2.Normal-dose of hesperidin(Hesd-20,20 mg/kg)or high-dose of hesperidin(Hesd-200,200 mg/kg)single administration did not show any significant effect on pharmacokinetics,tissue distribution and biliary excretion of CPT-11 and SN-38 in rats,indicating that hesperidin may have no direct interaction with CPT-11 and SN-38.3.Multiple doses of hesperidin treatment significantly reduced biliary excretion of CPT-11 and SN-38 and markedly increased the concentration of CPT-11 and SN-38 in liver and colon tissue,the concentration of CPT-11 and SN-38 in intestinal contents was also decreased after hesperidin treatment,which may result from the inhibited biliary excretion of CPT-11 and SN-38 by hesperidin.4.Multiple dose of hesperidin treatment significantly decreased hepatic protein expression of Mrp2 which is considered to be the main transporter responsible for biliary excretion of CPT-11 and SN-38,but the protein expression of Bcrp,Mdr1 and Oatp in liver did not show any alteration.5.Multiple dose of hesperidin treatment mild,but not significantly improved the body weight,diarrhea score,and colonic pathological alteration in CIDD model rats.Further investigation was still needed to explore the protective effect of hesperidin on CIDD.Conclusions: Multiple dose of hesperidin treatment significantly reduced the hepatic Mrp2 expression,and thus increased systemic distribution of CPT-11 and SN-38,which further reduced the biliary excretion of CPT-11 and SN-38,especially the concentration of intestinal contents,and finally reduced the intestinal exposure of SN-38.These results indicating that hesperidin may have protective potential in reducing the incidence and severity of CIDD.