Department of Pharmacy, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, 100045, China.
School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing 210009, China.
Anal Methods. 2022 Jun 16;14(23):2293-2303. doi: 10.1039/d2ay00533f.
Ruxolitinib (RUX), a small molecule inhibitor of JAK1/JAK2, has been identified as the possible novel targeted agent for the treatment of hemophagocytic lymphohistiocytosis (HLH). However, due to the lack of randomized clinical trials (RCTs), it is extremely difficult to determine the effective therapeutic dose for RUX in HLH patients, especially in pediatric patients. At the same time, the clinical response of pediatric patients to RUX varies greatly among individuals according to several case reports. Therefore, it is imperative to investigate the pharmacokinetic and pharmacodynamic characteristics of RUX in HLH children, and this must be based on a satisfactory method to determine the concentration of RUX. Owing to several limits of published analytical methods, herein, we describe a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) method for monitoring RUX in children's plasma samples. The protein precipitation method using methanol was used for sample cleanup. The analytes were separated by gradient elution in which 2.0 mM ammonium acetate in distilled water and acetonitrile were used as mobile phases. In the positive electrospray ionization (ESI+) mode, the / 307.1 → 186.0 and 316.1 → 185.9 ion pair transitions of RUX and RUX-d9 were used for the qualitative and quantitative analysis, respectively. The calibration curves of RUX were linear in the concentration range from 0.5 to 400 ng mL. The intra- and inter-batch precision, accuracy, recovery, dilution completeness, and stability of this method were all within acceptable standards, and no matrix effects or residues were found. This method was successfully applied to the clinical pharmacokinetic study of RUX in 32 children with HLH. The pharmacokinetic parameters of HLH children after a single dose of RUX and the steady state plasma concentration after multiple administrations were proposed through this method. Most importantly, it was found that the age and serum creatinine (SCr) of children with HLH had a significant and complex impact on the process of RUX after the single as well as multiple administrations of RUX.
芦可替尼（RUX）是一种小分子 JAK1/JAK2 抑制剂，已被确定为噬血细胞性淋巴组织细胞增生症（HLH）治疗的潜在新型靶向药物。然而，由于缺乏随机临床试验（RCT），很难确定 RUX 在 HLH 患者中的有效治疗剂量，尤其是在儿科患者中。同时，根据几项病例报告，儿科患者对 RUX 的临床反应因人而异。因此，有必要研究 RUX 在 HLH 儿童中的药代动力学和药效动力学特征，而这必须基于一种确定 RUX 浓度的令人满意的方法。由于已发表的分析方法存在一些局限性，因此，我们在此描述了一种用于监测儿童血浆样品中 RUX 的新型液相色谱串联质谱（LC-MS/MS）方法。使用甲醇进行蛋白沉淀法用于样品净化。在梯度洗脱中，使用 2.0 mM 氨乙酸在蒸馏水中和乙腈作为流动相分离分析物。在正电喷雾电离（ESI+）模式下，使用 RUX 和 RUX-d9 的 307.1→186.0 和 316.1→185.9 离子对转换对 RUX 进行定性和定量分析。RUX 的校准曲线在 0.5 至 400 ng mL 的浓度范围内呈线性。该方法的批内和批间精密度、准确度、回收率、稀释完整性和稳定性均在可接受范围内，未发现基质效应或残留。该方法成功应用于 32 例 HLH 儿童的 RUX 临床药代动力学研究。通过该方法提出了 HLH 儿童单次 RUX 给药后的药代动力学参数和多次给药后的稳态血浆浓度。最重要的是，研究发现 HLH 儿童的年龄和血清肌酐（SCr）对 RUX 单次和多次给药后的药代动力学过程有显著且复杂的影响。
