| Objective:To investigate the anti-tumor activity of Copen in a series of tumor cell lines in vitro and in vivo. To investigate the pharmacokinetics of Copen in rats, and obtain the main pharmacokinetic parameters. And thus provide support for the further development of this drug.Methods:1. MTT method was used to detect the proliferation inhibition effect of Copen on14types of tumor cell lines.2. MTT method was used to detect the proliferation inhibition effect of Copen combined with taxol/topotecan/cisplatin on HCT-116tumor cell.3. The in vivo effect was determined with5types of tumor xenograft model in nude mice. Relative tumor volume, tumor weight and inhibition rate were evaluated respectively after the mice were sacrificed.4. A validated LC-MS/MS assay was established to determinate the plasma concentration of Copen in rats. 5. The pharmacokinetic characteristic of Copen in rats was investigated. The relationship between AUC-dose/Cmax-dose and gender differences in the main pharmacokinetic parameters was analysed statistically.Results:1. Copen exhibited proliferation inhibition effect on the different tumor cell lines with IC50values mostly between300-1000μM. Copen exerted a weaker inhibition effect on the14types of tumor cells in contrast with the positive-control group (taxol). And Copen showed a relatively obvious proliferation inhibition effect on95-D, Bel-7402, MDA-MB-231, PC-3and HL-60tumor cells with the IC50value less than500μM.2. No obvious synergic effect was observed when Copen (800μM、400μM) was used with taxol (18.25-600ng/mL) or topotecan (0.15625～5μmol/mL). However Copen (800μM) could significantly improved the proliferation inhibition effect on HCT-116tumor cells in combination with cisplatin (0.5,1.0,2.0μg/mL).3. No obvious growth inhibition on SMMC-7721, Bel-7402, HepG2and95-D xenografts in nude mice was observed when Copen was administered intravenously with the test dose100-25mg/kg. However Copen (100or50mg/kg) could significantly inhibit the growth of HCT-116xenografts in nude mice when administered intravenously.4. A sensitive and specific LC-MS/MS was developed with linear range of0.05158～20.63μg·mL-1and1.01～504.76μg·mL-1respectively. Intra-day and inter-day precision values were within11.3%. The overall recovery for Copen was greater than65%. This validated method was successfully applied to the pharmacokinetic study.5. When Copen was administered orally with the test dose25mg/kg,50mg/kg and100mg/kg,the main pharmacokinetics parameters were as follows:t1/2were (302.16± 169.90) min,(243.80±112.71) min and (196.55±66.44) min, respectively. Cmax were (13.39±10.17) mg-L"1,(21.96±25.84) mg·L-1and (28.35±10.24) mg·L-1, respectively. Tmax were (10.00±3.69) min,(9.17±3.59) min and (14.17±24.01) min, respectively. AUC0-t were (729.59±4337.80) mg·L-1·h,(1468.60±920.76) mg·L-1·h and (5167.31±2842.48) mg·L-1·h, respectively.When Copen was administered intravenously with the test dose25mg/kg,50mg/kg and100mg/kg, the main pharmacokinetics parameters were as follows:t1/2were (296.49±571.77) min,(158.07±110.36) min and (229.05±260.11) min, respectively. Cmax were (339.34±290.40) mg·L-1,(553.68±539.87) mg·L-1and (1451.58±1108.34) mg·L-1, respectively. Tmax were (15.00±17.06) min,(6.25±3.11) min and (6.67±3.26) min, respectively. AUC0-t were (24545.80±21102.82) mg·L-1·min,(54342.71±76451.41) mg·L-1·min and (68653.62±37177.46) mg·L-1·min, respectively.As statistical analysis showed, a positive relationship was found at doses from25to100mg/kg after a single-dose orally administered or intravenous infusion. Gender differences were not observed in the pharmacokinetics of Copen when administered intravenously. However, gender differences were observed in Cmax (P<0.05) when Copen was administered orally.Conclusion:1. Copen showed a relatively strong proliferation inhibition effect on95-D, Bel-7402, MDA-MB-231, PC-3and HL-60tumor cells.2. Copen could significantly improve the proliferation inhibition on HCT-116tumor cells in combination with cisplatin.3. Copen (100or50mg/kg) could significantly inhibit the growth of HCT-116xenografts in nude mice when administered intravenously.4. A sensitive and selective LC-MS/MS method was developed, validated, and successfully applied to the pharmacokinetic study.5. A positive relationship was found at doses from25to100mg/kg after a single-dose orally or intravenously administered. Gender difference has a greater impact on absorption compared with elimination. |
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