Effects Of Wuzhi Capsule On The Exposure Of Apatinib And The Anti-Tumor Activity Of Sch.B For Apatinib

Part one Effect of Wuzhi Capsule on the pharmacokinetics of apatinib in ratsObjective:To establish a determination method of apatinib concentration in rat plasma by LC-MS/MS;and study the effects of Wuzhi Capsule with single dose and multiple dose on the pharmacokinetic of apatinib in rats.Methods:24 male Sprague-Dawley?SD?rats were randomly divided into four groups:control group I(apatinib 50 mg·kg^-1),treatment group I(apatiinb 50 mg·kg^-1 and Wuzhi Capsule 450 mg·kg^-1),control group II?apatinib after given normal saline once a day for 7 d?,treatment group II?apatinib after given Wuzhi Capsule once a day for 7 d?.Blood samples were collected at different time points and analyzed the blood concentration of apatinib by LC-MS/MS.After protein precipitation with methanol,samples were chromatographed through a Waters XBridge BEH C18 column.The mobile phase was acetonitrile and water?including 0.1%formic acid?.For the detection of apatinib and carbamazepine?IS?,the mass spectrometer was set to positive ionization mode,The multiple reaction monitoring?MRM?mode was used to quantify the analytes and IS.Non-compartmental analysis was performed to calculate the pharmacokinetic parameters using DAS 2.1.Results:A full evaluation was made.A good linearity was obtained over the range of 2²000 ng·mL^-1,and all the indeses of the methodological investigations meet the requirements.The pharmcokinetic result showed that compared with control group I,C_max and AUC_0-24h of apatinib in treatment group I increased by about 102%and 315%,respectively,while the CL_z decreased by about 78%.Compared with control group II,C_max and AUC_0-24h-24h in treatment group II increased by about 52%and 169%,respectively,while the CL_z decreased by about 61%.Conclusions:In this study,a sensitive and specific UPLC-MS/MS method to quantify apatinib in rat plasma samples was developed.And this method was applied to study the effect of Wu-zhi capsule on apatinib pharmacokinetics.Wuzhi Capsule with single dose and multiple dose could significantly increase the exposure of apatinib in rats.Part two The synergistic anti-tumor effect of schisandrin B and apatinibObjective:To study the synergistic anti-tumor effect and mechanism of Sch.B combined with apatinib.Methods:The CCK-8 assay was used to assess the proliferation of SGC-7901 cells.The wound healing assay was used to evaluate the effect of apatinib and Sch.B individually and in combination on the migration of SGC-7901 cells.Transwell co-culture chambers were used to evaluate the effect of apatinib and Sch.B on SGC-7901 cell invasion.The flow cytometry was used to detect the cell cycle and percentage of apoptosis of SGC-7901 cell lines and observed the impact of different medications on cell lines.Western blot was used to investigate the effect of apatinib,Sch.B and combined apatinib with Sch.B on the expression of MMP-9,Bax,Caspase-9,Caspase-12.Results:The results of CCK-8 and cell cycle assay showed that the combination of Sch.B and apatinib significantly enhanced the inhibition on SGC-7901 cell proliferation by means of arresting G0/G1 phase.The wound-healing and transwell invasion assay showed that this combination enhanced inhibition on SGC-7901 cells migration and invasion.The result of flow cytometry showed that the combination of Sch.B and apatinib can significantly increase the apoptosis of SGC-7901 cells.Moreover the protein espression of MMP-9,Bax,Caspase-9,and Caspase-12 detected by western blot also enhanced when treat with the combination with apatinib and Sch.B.Conclusions:Combined with apatinib and Sch.B enhanced the anti-cancer effect of SGC-7901 cells on proliferation,migration,invasion and apoptosis in SGC-7901 cells,and this result correlated with the expression of MMP-9,Bax,Caspase-9,Caspase-12.