五酯胶囊对大鼠体内甲磺酸阿帕替尼药动学行为的影响
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摘要
目的:建立测定大鼠血浆中甲磺酸阿帕替尼浓度的方法,并考察单次和多次给予五酯胶囊对大鼠体内甲磺酸阿帕替尼药动学行为的影响。方法:采用液相色谱-串联质谱法(LC-MS/MS)检测大鼠血浆中甲磺酸阿帕替尼的浓度。以卡马西平为内标,色谱柱为Waters XBridge BEH C_(18),流动相为乙腈-0.1%甲酸溶液(45∶55,V/V),流速为0.3 mL/min,柱温为40℃,进样器温度为15℃,进样量为2μL;采用电喷雾离子源,以多反应监测模式进行正离子扫描,用于定量分析的离子对质荷比分别为398.1→212.1(甲磺酸阿帕替尼)、237.2→194.2(内标)。将大鼠随机分为对照组Ⅰ、观察组Ⅰ、对照组Ⅱ、观察组Ⅱ,每组6只。对照组Ⅰ大鼠单次灌胃甲磺酸阿帕替尼(50 mg/kg,下同);观察组Ⅰ大鼠灌胃五酯胶囊(450 mg/kg,下同),10 min后再灌胃甲磺酸阿帕替尼;对照组Ⅱ大鼠灌胃生理盐水,每日1次,连续7 d后,再单次灌胃甲磺酸阿帕替尼;观察组Ⅱ大鼠灌胃五酯胶囊,每日1次,连续7 d后,再单次灌胃甲磺酸阿帕替尼。分别于灌胃甲磺酸阿帕替尼后0.25、0.5、1.0、2.0、2.5、3.0、4.0、6.0、8.0、12.0、24.0 h时自大鼠眼内眦静脉丛采血进行测定。分别采用DAS 2.1软件和t检验计算和比较各组大鼠的药动学参数。结果:甲磺酸阿帕替尼检测血药浓度的线性范围为2～2 000 ng/mL,定量下限为2 ng/mL;日内、日间RSD均小于10%,准确度为94.93%～104.68%;基质效应不影响待测物的定量分析。与对照组Ⅰ比较,观察组Ⅰ大鼠的c_(max)、AUC_(0-24h)、AUC_(0-∞)均显著升高,CLZ显著降低(P<0.05);与对照组Ⅱ比较,观察组Ⅱ大鼠的AUC_(0-24 h)、AUC_(0-∞)均显著升高,CLZ显著降低(P<0.05);与观察组Ⅰ比较,观察组Ⅱ大鼠的AUC_(0-24h)、AUC_(0-∞)均显著降低(P<0.05)。结论:本研究建立的LC-MS/MS法灵敏度高、专属性强,可用于大鼠血浆中甲磺酸阿帕替尼浓度的测定及药动学研究。五酯胶囊可影响大鼠体内甲磺酸阿帕替尼的药动学行为,且多次给予五酯胶囊的影响弱于单次给药。
OBJECTIVE:To establish a method for the concentration determination of apatinib mesylate in plasma of rats,and to investigate the effects of single and multiple administration of Wuzhi capsules on the pharmacokinetic behavior of apatinib mesylate in rats. METHODS: LC-MS/MS method was used to detect the plasma concentration of apatinib mesylate in rats. Using carbamazepine as internal standard,the determination was performed on Waters XBridge BEH C_(18) column with mobile phase consisted acetonitrile-0.1% formic acid solution(45 ∶ 55,V/V)at the flow rate of 0.3 mL/min. The column temperature was 40 ℃.The temperature of injector was 15 ℃,and the sample size was 2 μL. ESI was used for positive ion scanning in MRM mode. The ion pairs m/z used for quantitative analysis were 398.1→212.1(apatinib mesylate)and 237.2→194.2(internal standard). The rats were randomly divided into control group Ⅰ,observation group Ⅰ,control group Ⅱ,observation group Ⅱ,with 6 rats in each group. Control group Ⅰ were given single administration of apatinib mesylate intragastrically(50 mg/kg,similarly hereinafter).Observation group Ⅰ was given Wuzhi capsules intragastrically(450 mg/kg,similarly hereinafter),and then 10 min later given apatinib mesylate intragastrically. Control group Ⅱ was given normal saline intragastrically,once a day,for consecutive 7 d,and then were given single administration of apatinib mesylate. Observation group Ⅱ was given Wuzhi capsules intragastrically,once a day,for consecutive 7 d,and then 10 min later were given single administration of apatinib mesylate. The blood samples were collected from intraocular canthus vein plexus and determined 0.25,0.5,1.0,2.0,2.5,3.0,4.0,6.0,8.0,12.0,24.0 h after intragastric administration,respectively. Pharmacokinetic parameters were apatinib mesylate were calculated and compared among those groups by using DAS 2.1 software and t-test. RESULTS:The linear range of apatinib mesylate were 2-2 000 ng/mL. The lower limit of quantitation was 2 ng/mL. RSDs of intra-day and inter-day were all lower than 10%,and the accuracy were 94.93%-104.68%. Matrix effect did not affect the quantitative analysis of the substance to be measured. Compared with control group Ⅰ,cmax,AUC_(0-24) hand AUC(0-∞) of observation group Ⅰ were increased significantly, CLZwas decreased significantly(P<0.05). Compared with control group Ⅱ,AUC_(0-24) hand AUC_(0-∞) of observation group Ⅱ were increased significantly,and CLZ was decreased significantly(P<0.05). Compared with observation group Ⅰ,AUC_(0-24) hand AUC_(0-∞) of observation group Ⅱ were decreased significantly(P<0.05). CONCLUSIONS:Established LC-MS/MS method is sensitive and specific,and can be used for the concentration determination of apatinib mesylate in plasma of rats. Wuzhi capsules can influence in vivo pharmacokinetic behavior of apatinib mesylate in rats. The effect of multiple administration of Wuzhi capsules is weaker than that of single administration.
OBJECTIVE:To establish a method for the concentration determination of apatinib mesylate in plasma of rats,and to investigate the effects of single and multiple administration of Wuzhi capsules on the pharmacokinetic behavior of apatinib mesylate in rats. METHODS: LC-MS/MS method was used to detect the plasma concentration of apatinib mesylate in rats. Using carbamazepine as internal standard,the determination was performed on Waters XBridge BEH C_(18) column with mobile phase consisted acetonitrile-0.1% formic acid solution(45 ∶ 55,V/V)at the flow rate of 0.3 mL/min. The column temperature was 40 ℃.The temperature of injector was 15 ℃,and the sample size was 2 μL. ESI was used for positive ion scanning in MRM mode. The ion pairs m/z used for quantitative analysis were 398.1→212.1(apatinib mesylate)and 237.2→194.2(internal standard). The rats were randomly divided into control group Ⅰ,observation group Ⅰ,control group Ⅱ,observation group Ⅱ,with 6 rats in each group. Control group Ⅰ were given single administration of apatinib mesylate intragastrically(50 mg/kg,similarly hereinafter).Observation group Ⅰ was given Wuzhi capsules intragastrically(450 mg/kg,similarly hereinafter),and then 10 min later given apatinib mesylate intragastrically. Control group Ⅱ was given normal saline intragastrically,once a day,for consecutive 7 d,and then were given single administration of apatinib mesylate. Observation group Ⅱ was given Wuzhi capsules intragastrically,once a day,for consecutive 7 d,and then 10 min later were given single administration of apatinib mesylate. The blood samples were collected from intraocular canthus vein plexus and determined 0.25,0.5,1.0,2.0,2.5,3.0,4.0,6.0,8.0,12.0,24.0 h after intragastric administration,respectively. Pharmacokinetic parameters were apatinib mesylate were calculated and compared among those groups by using DAS 2.1 software and t-test. RESULTS:The linear range of apatinib mesylate were 2-2 000 ng/mL. The lower limit of quantitation was 2 ng/mL. RSDs of intra-day and inter-day were all lower than 10%,and the accuracy were 94.93%-104.68%. Matrix effect did not affect the quantitative analysis of the substance to be measured. Compared with control group Ⅰ,cmax,AUC_(0-24) hand AUC(0-∞) of observation group Ⅰ were increased significantly, CLZwas decreased significantly(P<0.05). Compared with control group Ⅱ,AUC_(0-24) hand AUC_(0-∞) of observation group Ⅱ were increased significantly,and CLZ was decreased significantly(P<0.05). Compared with observation group Ⅰ,AUC_(0-24) hand AUC_(0-∞) of observation group Ⅱ were decreased significantly(P<0.05). CONCLUSIONS:Established LC-MS/MS method is sensitive and specific,and can be used for the concentration determination of apatinib mesylate in plasma of rats. Wuzhi capsules can influence in vivo pharmacokinetic behavior of apatinib mesylate in rats. The effect of multiple administration of Wuzhi capsules is weaker than that of single administration.
引文
[1]GENG R,LI J.Apatinib for the treatment of gastric cancer[J].Expert Opin Pharmacother,2015,16(1):117-122.
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[4]杨福泰,李向东,黄巧云.五酯胶囊治疗各种病因所致血清转氨酶升高120例[J].现代中西医结合杂志,2004,13(8):1044.
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[6]杨燕,辛华雯,刘飞,等.五酯胶囊对他克莫司增效作用与CYP3A5*3基因多态性的相关性研究[J].中国药房,2017,28(5):581-585.
[7]陈揭剑,梅长林.五酯胶囊对特发性膜性肾病患者环孢素A全血浓度的影响研究[J].中国全科医学,2015,18(7):842-845.
[8]国家药典委员会.中华人民共和国药典:四部[S].2015年版.北京:中国医药科技出版社,2015:363-368.
[9]GILLESPIE WR.非房室和房室模型分析在临床药动学的比较[J].国外医学药学分册,1992,19(1):40-43.
[10]冯健男,杜守颖,白洁,等.生物样品前处理的研究进展[J].中国中药杂志,2014,39(21):4143-4148.
[11]乔丽曼,王豪,楼旦,等.超高液相色谱-质谱法检测人血浆中格列美脲和氟西汀及其代谢产物[J].中国临床药理学杂志,2015,31(18):1873-1876.
[12]黄继汉,黄晓晖,陈志扬,等.药理试验中动物间和动物与人体间的等效剂量换算[J].中国临床药理学与治疗学,2004,9(9):1069-1072.
[13]位华.五酯胶囊的药代动力学及与他克莫司相互作用研究[D].上海:第二军医大学,2010.
[14]宋敏,谭鸿毅,谭志荣,等.单剂量和多剂量口服布洛伪麻那敏干混悬剂在健康人体的药代动力学[J].中国临床药理学杂志,2010,26(1):28-32、36.
[15]王玉浩,张雪,周小庭,等.舒尼替尼和雷米普利联合用药在大鼠体内的药代动力学相互作用[J].中国药科大学学报,2017,48(1):60-65.
[16]张汝波.华中五味子的化学成分研究[D].昆明:昆明理工大学,2007.
[17]李伟,刘亚丽,宋永贵,等.UPLC-Q-TOF-MSE结合OPLS-DA模式快速鉴定南、北五味子化学成分与识别差异标志物[J].中草药,2015,46(15):2212-2218.
[18]吴育晶,程能能,胡卓汉,等.五味子及木脂素成分对肝CYP3A酶活性的影响[J].中国临床药学杂志,2007,16(5):267-271.
[19]王卓,王湛博,程亚楠,等.五味子乙素对CYP450药物代谢酶诱导作用的体外研究[J].药学研究,2017,36(10):559-564.
[20]陈倩,吴育晶,程能能,等.五味子提取物对大鼠肝CYP3A酶的双重作用[J].药学学报,2010,45(9):1194-1198.
[21]吴笑春,辛华雯,李罄,等.五酯胶囊对健康受试者他克莫司药动学的影响[J].中国新药杂志,2007,16(8):647-650.
[22]黄琪,裴奇,周于禄,等.五酯胶囊对拉米夫定人体内药物动力学的影响[J].中南药学,2007,5(5):461-463.
[23]石庆玲.五酯胶囊对阿托伐他汀药动学的影响[D].上海:复旦大学,2014.
[24]XUE XP,QIN XL,XU C,et al.Effect of Wuzhi tablet(Schisandra sphenanthera extract)on the pharmacokinetics of cyclosporin A in rats[J].Phytother Res,2013,27(8):1255-1259.
[2]LI J,QIN S,XU J,et al.Randomized,double-blind,placebo-controlled phaseⅢtrial of apatinib in patients with chemotherapy-refractory advanced or metastatic adenocarcinoma of the stomach or gastroesophageal junction[J].JClin Oncol,2016,34(13):1448-1454.
[3]LIU X,QIN S,WANG Z,et al.Early presence of anti-angiogenesis-related adverse events as a potential biomarker of antitumor efficacy in metastatic gastric cancer patients treated with apatinib:a Cohort study[J].J Hematol Oncol,2017.DOI:10.1186/s13045-017-0521-0.
[4]杨福泰,李向东,黄巧云.五酯胶囊治疗各种病因所致血清转氨酶升高120例[J].现代中西医结合杂志,2004,13(8):1044.
[5]DING J,CHEN X,GAO Z,et al.Metabolism and pharmacokinetics of novel selective vascular endothelial growth factor receptor-2 inhibitor apatinib in humans[J].Drug Metab Dispos,2013,41(6):1195-1210.
[6]杨燕,辛华雯,刘飞,等.五酯胶囊对他克莫司增效作用与CYP3A5*3基因多态性的相关性研究[J].中国药房,2017,28(5):581-585.
[7]陈揭剑,梅长林.五酯胶囊对特发性膜性肾病患者环孢素A全血浓度的影响研究[J].中国全科医学,2015,18(7):842-845.
[8]国家药典委员会.中华人民共和国药典:四部[S].2015年版.北京:中国医药科技出版社,2015:363-368.
[9]GILLESPIE WR.非房室和房室模型分析在临床药动学的比较[J].国外医学药学分册,1992,19(1):40-43.
[10]冯健男,杜守颖,白洁,等.生物样品前处理的研究进展[J].中国中药杂志,2014,39(21):4143-4148.
[11]乔丽曼,王豪,楼旦,等.超高液相色谱-质谱法检测人血浆中格列美脲和氟西汀及其代谢产物[J].中国临床药理学杂志,2015,31(18):1873-1876.
[12]黄继汉,黄晓晖,陈志扬,等.药理试验中动物间和动物与人体间的等效剂量换算[J].中国临床药理学与治疗学,2004,9(9):1069-1072.
[13]位华.五酯胶囊的药代动力学及与他克莫司相互作用研究[D].上海:第二军医大学,2010.
[14]宋敏,谭鸿毅,谭志荣,等.单剂量和多剂量口服布洛伪麻那敏干混悬剂在健康人体的药代动力学[J].中国临床药理学杂志,2010,26(1):28-32、36.
[15]王玉浩,张雪,周小庭,等.舒尼替尼和雷米普利联合用药在大鼠体内的药代动力学相互作用[J].中国药科大学学报,2017,48(1):60-65.
[16]张汝波.华中五味子的化学成分研究[D].昆明:昆明理工大学,2007.
[17]李伟,刘亚丽,宋永贵,等.UPLC-Q-TOF-MSE结合OPLS-DA模式快速鉴定南、北五味子化学成分与识别差异标志物[J].中草药,2015,46(15):2212-2218.
[18]吴育晶,程能能,胡卓汉,等.五味子及木脂素成分对肝CYP3A酶活性的影响[J].中国临床药学杂志,2007,16(5):267-271.
[19]王卓,王湛博,程亚楠,等.五味子乙素对CYP450药物代谢酶诱导作用的体外研究[J].药学研究,2017,36(10):559-564.
[20]陈倩,吴育晶,程能能,等.五味子提取物对大鼠肝CYP3A酶的双重作用[J].药学学报,2010,45(9):1194-1198.
[21]吴笑春,辛华雯,李罄,等.五酯胶囊对健康受试者他克莫司药动学的影响[J].中国新药杂志,2007,16(8):647-650.
[22]黄琪,裴奇,周于禄,等.五酯胶囊对拉米夫定人体内药物动力学的影响[J].中南药学,2007,5(5):461-463.
[23]石庆玲.五酯胶囊对阿托伐他汀药动学的影响[D].上海:复旦大学,2014.
[24]XUE XP,QIN XL,XU C,et al.Effect of Wuzhi tablet(Schisandra sphenanthera extract)on the pharmacokinetics of cyclosporin A in rats[J].Phytother Res,2013,27(8):1255-1259.