新型IDO1/TDO抑制剂SHR9146的有关物质检测

Detection of related substances of SHR9146—a novel IDO1/TDO dual inhibitor

目的:建立高效液相色谱法进行肿瘤免疫治疗的新型高选择性吲哚胺2,3-双加氧酶1(IDO1)/色氨酸2,3-加氧酶(TDO)抑制剂(S)-2-(4-(4-(4-(6-氟-5H-咪唑并[5,1-a]异吲哚-5-基)哌啶-1-基)苯基)-1H-吡唑-1-基)乙醇(SHR9146)的有关物质检测。方法:采用Kromasil C_(18)(250 mm×4.6 mm,5μm)色谱柱,以0.01 mol·L^(-1)磷酸二氢钾缓冲溶液(含0.3%三乙胺,磷酸调节pH至3.0)(A)-乙腈(B)为流动相,梯度洗脱,流速1.0 mL·min^(-1),紫外检测波长270 nm。结果:SHR9146与各有关物质的色谱分离良好,色谱响应在0.01~5.0μg·mL^(-1)浓度范围内线性关系良好(r>0.999),检测限和定量限分别约为21.95和73.16 ng·mL^(-1)。利用上述方法,对3批产品的有关物质进行分析,并采用加校正因子的主成分自身对照法计算含量,杂质SHR9146Z-399含量在0.46%~0.60%,SHR9146Z-333含量不超过0.11%,其他单个杂质含量均不超过0.10%,有关物质含量总和低于1.0%。结论:建立的HPLC-UV方法专属性好,检测灵敏度高,可用于SHR9146原料药的质量控制。

Objective:To establish an HPLC method for the analysis of related substances in(S)-2-(4-(4-(4-(6-fluoro-5H-imidazo[5,1-a]isoindol-5-yl)piperidin-1-yl)phenyl)-1H-pyrazol-1-yl)ethan-1-ol(SHR9146),a novel high selective indoleanmine 2,3-dioxygenase(IDO1)/tryptophan 2,3-dioxygenaes(TDO)inhibitor used for tumor immunotherapy.Methods:The chromatographic separation was performed on a Kromasil C_(18)column(250 mm×4.6 mm,5μm)with gradient elution by 0.01 mol·L^(-1)monopotassium phosphate buffer(containing 0.1% triethylamine and adjusted pH to 3.0 with phosphate acid)and acetonitrile as mobile phase at a flow rate of 1.0 mL·min^(-1),and the UV detection wavelength was set at 270 nm.Results:The peaks of SHR9146 and related substances could be separated completely.UPLC-UV analysis could achieve a good linear regression in the concentration range of 0.01-5.0μg·mL^(-1)(r>0.999).The detection limit and quantification limit of SHR9146 were 21.95 and 73.16 ng·mL^(-1)respectively.Using the HPLC method,the related substances of three batches were detected and calculated by using principal component self-control with correct factor,the total impurities were less than 1.0% with impurity SHR9146Z-399 in the range of 0.46%-0.60% and SHR9146Z-333 and other single impurity not more than 0.11% and 0.10% respectively.Conclusion:The established method has good specificity and high sensitivity,and can be used for the quality control of SHR9146.