16种具有P-gp抑制作用的上市药物对吉非替尼口服生物利用度和脑通透性的影响
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摘要
采用LC-MS/MS法研究16种具有P-gp抑制作用的上市药物对吉非替尼口服生物利用度和脑通透性的影响。对照组ICR小鼠灌胃给予CMC-Na混悬液和吉非替尼混悬液,抑制剂组ICR小鼠分别灌胃给予16种上市药物混悬液和吉非替尼混悬液,血浆样品和脑匀浆样品经乙腈沉淀后,用LC-MS/MS法测定药物浓度。结果发现16种药物中利托那韦能够明显增加吉非替尼的口服生物利用度,吉非替尼血药浓度-时间曲线下面积(AUC)增加了2倍,同时增加了吉非替尼的脑暴露量,但是没有增加吉非替尼的脑通透性;部分其他药物也能增加吉非替尼的血浆AUC,不能增加吉非替尼的脑通透性;游离分数校正脑浓度发现对照组和给予利托那韦后,脑内游离药物浓度均不能达到体外抑制非小细胞肺癌(NSCLC)细胞生长的IC_(50)。本研究结果表明,临床剂量的16种具有P-gp抑制作用的上市药物虽然能够增加吉非替尼的口服生物利用度,但是不能特异性提高脑组织暴露量,需要研发特异性更强和更安全的P-gp抑制剂;用脑部药物游离分数校正脑浓度后,临床前研究发现脑暴露量不足可能是吉非替尼治疗脑转移疗效不理想的原因之一。
To investigate the effects of clinical P-glycoprotein inhibitors on oral bioavailability and brain penetration of gefitinib,16 inhibitors and gefitinib were co-administered orally to ICR mice.The suspension of gefitinib and CMC-Na were co-administered to the control group.The suspension of gefitinib and clinical P-glycoprotein inhibitors were co-administered to the control group.Blood samples and brain homogenate samples were extracted by protein precipitation with acetonitrile and determinated by LC-MS/MS.It was found that ritonavir can significantly increase the oral bioavailability of gefitinib,and the area under the plasma concentration-time curves(AUC) of gefitinib was increased by 2 times;while brain exposure was increased,there was no increment in brain penetration.Some other drugs can also increase the plasma AUC of gefitinib,but can not enhance the brain penetration;After we corrected brain concentration with fraction of unbound drug in brain,it was found that the brain concentration of gefitinib in both control group and ritonavir group did not achieve the in vitro IC_(50) of inhibiting non-small cell lung cancer(NSCLC) cell growth.Our results suggest that clinical doses of the 16 clinical P-glycoprotein inhibitors can not specifically increase brain tissue exposure,more specific and safer P-gp inhibitors are required.After we corrected brain concentrations with fraction of unbound drug in brain,our preclinical studies found that insufficient brain exposure may be one of the reasons for the unsatisfactory efficacy of gefitinib in the treatment of brain metastases.
To investigate the effects of clinical P-glycoprotein inhibitors on oral bioavailability and brain penetration of gefitinib,16 inhibitors and gefitinib were co-administered orally to ICR mice.The suspension of gefitinib and CMC-Na were co-administered to the control group.The suspension of gefitinib and clinical P-glycoprotein inhibitors were co-administered to the control group.Blood samples and brain homogenate samples were extracted by protein precipitation with acetonitrile and determinated by LC-MS/MS.It was found that ritonavir can significantly increase the oral bioavailability of gefitinib,and the area under the plasma concentration-time curves(AUC) of gefitinib was increased by 2 times;while brain exposure was increased,there was no increment in brain penetration.Some other drugs can also increase the plasma AUC of gefitinib,but can not enhance the brain penetration;After we corrected brain concentration with fraction of unbound drug in brain,it was found that the brain concentration of gefitinib in both control group and ritonavir group did not achieve the in vitro IC_(50) of inhibiting non-small cell lung cancer(NSCLC) cell growth.Our results suggest that clinical doses of the 16 clinical P-glycoprotein inhibitors can not specifically increase brain tissue exposure,more specific and safer P-gp inhibitors are required.After we corrected brain concentrations with fraction of unbound drug in brain,our preclinical studies found that insufficient brain exposure may be one of the reasons for the unsatisfactory efficacy of gefitinib in the treatment of brain metastases.
引文
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[7] U.S.Food,Drug Administration.Drug development and drug interactions:table of substrates,inhibitors and inducers [EB/OL].(2016-09-26)[2018-12-19].https://www.fda.gov/drugs/developmentapprovalprocess/developmentresources/druginteractionslabeling/ucm093664.htm.
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[9] Anroopb N,Shery J.A simple practice guide for dose conversion between animals and human[J].J Basic Clin Pharm,2016,7(2):27-31.
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[11] Cross DaE,Ashton SE,Ghiorghiu S,et al.AZD9291,an irreversible EGFR TKI,overcomes T790M-mediated resistance to EGFR inhibitors in lung cancer[J].Cancer Discov,2014,4(9):1046-1061.
[12] Swaisland HC,Ranson M,Smith RP,et al.Pharmacokinetic drug interactions of gefitinib with rifampicin,itraconazole and metoprolol[J].Clin Pharmacokinet,2005,44(10):1067-1081.
[2] Socinski MA,Evans T,Gettinger S,et al.Treatment of stage IV non-small cell lung cancer diagnosis and management of lung cancer,3rd ed:American college of chest physicians evidence-based clinical practice guideliness[J].Chest,2013,143(5):E341-E368.
[3] Zhao LL,Zhang Y,Zhou JP,et al.Advances in research on drugs targeting non-small cell lung cancer [J].J China Pharm Univ(中国药科大学学报),2014,45(2):136-144.
[4] Ceresoli GL,Cappuzzo F,Gregorc V,et al.Gefitinib in patients with brain metastases from non-small-cell lung cancer:a prospective trial[J].Ann Oncol,2004,15(7):1042-1047.
[5] Jackman DM,Holmes AJ,Lindeman N,et al.Response and resistance in a non-small-cell lung cancer patient with an epidermal growth factor receptor mutation and leptomeningeal metastases treated with high-dose gefitinib[J].J Clin Oncol,2006,24(27):4517-4520.
[6] Agarwal S,Sane R,Gallardo JL,et al.Distribution of gefitinib to the brain is limited by P-glycoprotein (ABCB1) and breast cancer resistance protein (ABCG2)-mediated active efflux[J].J Pharmacol Exp Ther,2010,334(1):147-155.
[7] U.S.Food,Drug Administration.Drug development and drug interactions:table of substrates,inhibitors and inducers [EB/OL].(2016-09-26)[2018-12-19].https://www.fda.gov/drugs/developmentapprovalprocess/developmentresources/druginteractionslabeling/ucm093664.htm.
[8] Lund M,Petersen TS,Dalhoff KP.Clinical implications of P-glycoprotein modulation in drug-drug interactions[J].Drugs,2017,77(8):859-883.
[9] Anroopb N,Shery J.A simple practice guide for dose conversion between animals and human[J].J Basic Clin Pharm,2016,7(2):27-31.
[10] Zeng QB,Wang JB,Cheng ZQ,et al.Discovery and evaluation of clinical candidate AZD3759,a potent,oral active,central nervous system-penetrant,epidermal growth factor receptor tyrosine kinase inhibitor[J].J Med Chem,2015,58(20):8200-8215.
[11] Cross DaE,Ashton SE,Ghiorghiu S,et al.AZD9291,an irreversible EGFR TKI,overcomes T790M-mediated resistance to EGFR inhibitors in lung cancer[J].Cancer Discov,2014,4(9):1046-1061.
[12] Swaisland HC,Ranson M,Smith RP,et al.Pharmacokinetic drug interactions of gefitinib with rifampicin,itraconazole and metoprolol[J].Clin Pharmacokinet,2005,44(10):1067-1081.