药物转运体相关遗传变异因素对核苷（酸）类似物抗HBV疗效的影响

The individual clinical efficacy is difference after anti-virus with nucleoside (acid) (NAs)at HBV patients. Drug was absorbed into the blood circulation through the digestive tract, then was transported into hepatocyte for inhibition of virus replication. The effects of genetic variation factor (Single Nucleotide Polymorphism, SNP) of transport (ENT, CNT, OCT and MRP4) of NAs at liver on steady state drug concentration of NAs in hepatocyte and the relationship between steady state drug concentration of NAs in hepatocyte and the SNP of transport on clinical efficacy are not yet clear explained. The relation of genetic factors of drug transport and the steady state drug concentration of NAs and clinical efficacy will be studied at the 1 year prospective follow-up study. NAs transporter gene polymorphism (Single Nucleotide Polymorphism, SNP) information and drug concentration will be tested by PCR method and Liquid chromatography tandem mass spectrometry (LC-MS/MS) method respectively. We research the relation between the SNP of transporter, the steady state drug concentration of NAs in hepatocyte and plasm and the clinical efficacy by logistic regression model. last, we check the function of the significant transporter SNP in vitro and get the SNP of NAs transporter which influences the steady state drug concentration of NAs in hepatocyte and clinical efficacy to guide the individual treatment.

乙肝患者对核苷（酸）类似物的药物疗效个体间存在差异。药物通过消化道吸收进入血液循环，然后进入肝脏进行抗病毒。肝脏核苷（酸）类似物相关的转运体（ENT、CNT、OCT 和MRP4）单核苷酸多态性（SNP）对个体核苷（酸）类似物稳态时肝脏细胞内药物浓度差异以及它们对药物疗效的影响尚未明确。本研究将对核苷（酸）类似物抗HBV治疗人群进行为期1年的随访，收集人口学资料、临床资料和生物样本。通过PCR方法和液相色谱串联质谱方法分别获得患者转运体SNP和体内药物浓度信息。构建logistic回归模型，研究转运体的SNP、核苷（酸）类似物稳态血药浓度、稳态时肝脏细胞内药物浓度和药物疗效之间的关系。并在体外对有意义的转运体SNP位点进行功能验证，筛选影响肝脏细胞内药物浓度和药物疗效的SNP位点，指导个体化治疗。

乙肝患者对核苷（酸）类似物的药物疗效个体间存在差异。药物通过消化道吸收进入血液循环，然后进入肝脏进行抗病毒。核苷（酸）类似物相关的转运体单核苷酸多态性（SNP）对个体核苷（酸）类似物稳态时药物浓度差异以及它们对药物疗效的影响尚未明确。最新一代核苷（酸）类似物替诺福韦艾拉酚胺半富马酸盐片（TAF）抗HBV效果较好，但是价格还是较高，为了降低TAF价格，仿制药成为首选，而TAF个体内变异尚未清楚，成为TAF仿制药完成生物等效性研究的难点。.本研究完成了核苷类似物甲磺酸帕拉德褔韦片在健康受试者中耐受性、药代动力学及其遗传变异研究；其在慢性乙型肝炎患者中治疗28天的耐受性、药代动力学、药效学及其遗传变异研究；其在慢性乙型肝炎患者中治疗48周疗效及其遗传变异研究。完成了替诺福韦艾拉酚胺半富马酸盐片（TAF）25mg随机、开放、单次给药、交叉健康人体生物等效性研究。.在健康受试者中发现ABCB4 rs2097937、CYP19A1 rs700518、CYP19A1 rs1062033、CYP2F1 rs305968基因位点多态性对甲磺酸帕拉德褔韦片在体内暴露量有影响，还可能导致药物高暴露后的不良反应；在慢性乙肝受试者治疗28天后，发现rs2517474位点的G等位基因、rs2517448位点的T基因、rs6457327位点的A基因与帕拉德福韦的暴露量AUC0-24显著相关，但是与疗效无明显相关。Rs6939511位点的G基因与帕拉德福韦代谢产物PMEA的暴露量AUC0-24和疗效指标HBV DNA下降量相关。包含rs6939511位点G基因人群的疗效优于不含G基因的人群(-3.37 VS -3.06 IU/mL分别在 A/G 和 A/A 基因型)；在慢性乙型肝炎患者治疗24周后，发现Rs6939511位点的G等位基因与帕拉德福韦代谢产物PMEA的稳态浓度-谷浓度显著相关，以及疗效指标HBV DNA下降量明显相关。.发现替诺福韦艾拉酚胺个体内变异高于替诺福韦，空腹条件下替诺福韦艾拉酚胺个体内变异在21.96-37.39%，推荐空腹生物等效性研究适合双交叉设计。餐后条件下下替诺福韦艾拉酚胺个体内变异约为46.1%，推荐餐后生物等效性研究适合三或者四交叉设计。.本研究阐明了帕拉德福韦的代谢特点、基因变异情况及其对药物浓度和疗效的影响。阐明了TAF的变异，促进国产TAF上市，造福百姓。