新型SHP2衍生物NC-55-122在大鼠体内的药动学研究及其体内代谢产物分析

Pharmacokinetic study of a novel SHP2 derivative NC-55-122 in rats and its metabolites in vivo

考察新型含Src同源2结构域蛋白酪氨酸磷酸酶(SHP2)蛋白抑制剂三唑并喹啉酮衍生物NC-55-122在大鼠体内的药代动力学特征,并在此基础上对化合物NC-55-122在大鼠体内的代谢产物进行分析。首先将大鼠随机分组,分别灌胃和静脉给予化合物NC-55-122,采集不同时间点的血样,以卡马西平为内标,采用超高效液相色谱-串联质谱法(UPLC-MS/MS)测定大鼠体内NC-55-122的浓度,并进行方法学的验证,用DAS 2.0软件计算主要的药代动力学参数,采用Graph Pad Prism 8.0.1软件绘制血药浓度-时间曲线;并同时建立超高液相色谱-四极杆-飞行时间质谱法(UPLC-Q-TOF/MS)对血浆样品进行分析,并运用UNIFI代谢产物预测软件分析经大鼠灌胃和静脉给药后的代谢产物。结果表明,化合物NC-55-122检测质量浓度的线性范围为1~1600 ng·m L^(-1),线性关系良好,在此线性范围内进行基质效应和提取回收率、精密度和准确度以及稳定性的考察,均符合生物分析要求;其次,药动学参数分析表明该化合物的口服生物利用度相对较低,仅为3.09%,但该化合物在体内吸收的速度较慢;最后通过分析离子流图以及结合UNIFI软件共推测出5个可能的代谢产物。本实验建立的检测方法灵敏度高,专属性强、快速、高效,适用于化合物NC-55-122在大鼠体内血药浓度的测定以及代谢产物的分析,为后期抗肿瘤药物NC-55-122的结构修饰以及成药性评价奠定基础。所有动物实验均经贵州医科大学实验动物伦理委员会批准(批准号:2200823)。

To investigate the pharmacokinetic characteristics and metabolites of Src homology 2 regioncontaining protein tyrosine phosphatase 2(SHP2)protein inhibitor in SD rats,a triazole quinolinone based derivative NC-55-122 was utilized.Firstly,rats were randomly divided into groups and given compound NC-55-122 intragastric and intravenous administration,respectively.Blood samples were collected at different time points.Taking carbmazepine as the internal standard,ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was used to determine the concentration of NC-55-122 in rats,and the methodology was verified.DAS 2.0 software was used to calculate the main pharmacokinetic parameters,and Graph Pad Prism8.0.1 software was used to plot the blood concentration-time curve.At the same time,UPLC-Q-TOF/MS was established to analyze plasma samples,and UNIFI metabolite prediction software was used to analyze metabolites after oral gavage and intravenous injection.The linear range of the mass concentration of compound NC-55-122was from 1 ng·m L~(-1)to 1600 ng·m L~(-1),and the linear relationship was good.The matrix effect,extraction recovery,precision,accuracy and stability were investigated in this linear range,which met the requirements of biological analysis.Secondly,the analysis of pharmacokinetic parameters showed that the oral bioavailability of the compound was low,with F%=3.09%,indicating that the compound was absorbed slowly in vivo.Finally,five possible metabolites were deduced by analyzing the ion flow diagram and combining UNIFI software.The detection method established in this experiment is highly sensitive,specific,rapid and efficient,which is suitable for the determination of the blood concentration of compound NC-55-122 in rats and the analysis of metabolites,and lays a foundation for the structural modification and druggability evaluation of the later anti-tumor drug NC-55-122.All animal experiments were approved by the Experimental Animal Ethics Committee of Guizhou Medical University(approval number:2200823).