Abstract：
Unecritinib (TQ-B3101) is a selective tyrosine kinase receptor inhibitor. In the study, in vitro metabolic experiments revealed that the hydrolysis of TQ-B3101 was mainly catalyzed by carboxylesterase 2 (CES2), followed by CES1. Next, a sensitive and reliable LC-MS/MS method was established for the simultaneous determination of TQ-B3101 and its metabolite crizotinib in rat plasma. To prevent in vitro hydrolysis of TQ-B3101, sodium fluoride, the CESs inhibitor at a concentration of 2 M, was immediately added after whole blood collection. Plasma samples were extracted by acetonitrile-induced protein precipitation method, and chromatographically separated on a Gemini C18 column (50 mm × 2.0 mm i.d., 5 μm) using gradient elution with a mobile phase of 0.1% formic acid and 5 mmol/L ammonium acetate with 0.1% formic acid. The retention times for TQ-B3101 and crizotinib were 2.61 and 2.38 min, respectively. The analytes were detected with tandem mass spectrometer by positive electrospray ionization, using the ion transitions at m/z 492.3 → 302.3 for TQ-B3101, m/z 450.3 → 260.3 for crizotinib, and m/z 494.0 → 394.3 for imatinib (internal standard). Method validation was conducted in the linear range of 1.00-800 ng/mL for the two analytes. The precision, accuracy and stabilities all met the acceptance criteria. The pharmacokinetic study indicated that TQ-B3101 was rapidly hydrolyzed to crizotinib with the elimination half-life of 1.11 h after a single gavage administration of 27 mg/kg to Sprague-Dawley rats, and the plasma exposure of TQ-B3101 was only 2.98% of that of crizotinib.
摘要：
Unecritinib(TQ-B3101)是一种选择性酪氨酸激酶受体抑制剂。在研究中,体外代谢实验表明，TQ-B3101的水解主要由羧酸酯酶2（CES2）催化，其次是CES1。接下来,建立了同时测定大鼠血浆中TQ-B3101及其代谢物克唑替尼浓度的LC-MS/MS方法。为了防止TQ-B3101，氟化钠的体外水解，浓度为2M的CESs抑制剂，在全血收集后立即添加。血浆样品采用乙腈诱导蛋白沉淀法提取,并在GeminiC18色谱柱上进行色谱分离（50mm×2.0mm内径，5μm），使用梯度洗脱，流动相为0.1％甲酸和5mmol/L乙酸铵，含0.1％甲酸。TQ-B3101和克唑替尼的保留时间分别为2.61和2.38分钟，分别。用串联质谱仪通过正电喷雾电离检测分析物，使用离子跃迁在m/z492.3→302.3对于TQ-B3101，m/z450.3→260.3对于克唑替尼，和m/z494.0→394.3伊马替尼（内标）。在1.00-800ng/mL的线性范围内对两种分析物进行方法验证。精度，准确性和稳定性均符合验收标准。药代动力学研究表明，对Sprague-Dawley大鼠单次灌胃给药后，TQ-B3101迅速水解为克唑替尼，消除半衰期为1.11h，TQ-B3101的血浆暴露量仅为克唑替尼的2.98%。
