Cui Xinge, Zheng Xin, Ren Jianwei, Liu Hongzhong, Jia Yuan, Wu Aiguo, Han Xiaohong
Clinical Pharmacology Research Center, State Key Laboratory of Complex Severe and Rare Diseases, NMPA Key Laboratory for Clinical Research and Evaluation of Drug, Beijing Key Laboratory of Clinical PK and PD Investigation for Innovative Drugs, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Jiangxi Jemincare Group Co., Ltd., Shanghai, China.
Front Pharmacol. 2023 Jan 13;14:1076046. doi: 10.3389/fphar.2023.1076046. eCollection 2023.
Etimicin is a fourth-generation aminoglycoside antibiotic. It has potent activity and low toxicity when employed for the treatment of Gram-negative and Gram-positive bacterial infections. The pharmacokinetics of etimicin in humans have not been elucidated completely. Two liquid chromatography-tandem mass spectrometry (LC-MS/MS) bioanalytical methods, without the use of any ion-pairing reagents, were developed and validated for the quantification of etimicin in human samples of serum and urine. Using a deuterated reagent as the internal standard, analytes in serum and urine samples were extracted by protein precipitation and dilution before LC-MS/MS analysis, respectively. For the two methods, chromatographic separations were undertaken under isocratic elution of water-ammonia solution-acetic acid (96:3.6:0.2, ) and methanol at 50%:50% and a flow rate of 0.35 ml/min within 5 min. A Waters XTerra MS C18 column (2.1 × 150 mm, 3.5 μm) and a column temperature of 40°C were chosen. A Sciex Qtrap 5500 mass spectrometer equipped with an electrospray ion source was used in both methods under multiple-reaction monitoring in positive-ion mode. The two methods showed good linearity, accuracy, and precision with high recovery and a minimal matrix effect in the range of 50.0-20000 ng/ml for serum samples and 50.0-10000 ng/ml for urine samples, respectively. Carry-over effects were not observed. Etimicin remained stable in human samples of serum or urine under the storage, preparation, and analytical conditions of the two methods. These two simple and reliable methods were applied successfully to a dose-escalation, phase I clinical trial of etimicin in Chinese healthy volunteers after intravenous administration of single and multiple doses. Based on these two methods we ascertained, for the first time, the comprehensive pharmacokinetics of etimicin in humans, which will be used for the exploration of the breakpoint research further.
依替米星是一种第四代氨基糖苷类抗生素。用于治疗革兰氏阴性菌和革兰氏阳性菌感染时，它具有强效活性且毒性低。依替米星在人体内的药代动力学尚未完全阐明。开发并验证了两种液相色谱 - 串联质谱（LC - MS/MS）生物分析方法，无需使用任何离子对试剂，用于定量人血清和尿液样本中的依替米星。分别以氘代试剂作为内标，血清和尿液样本中的分析物在LC - MS/MS分析前，分别通过蛋白沉淀和稀释进行提取。对于这两种方法，在水 - 氨溶液 - 乙酸（96:3.6:0.2）和50%甲醇与50%水等度洗脱、流速为0.35 ml/min、5分钟内的条件下进行色谱分离。选用沃特世XTerra MS C18柱（2.1×150 mm，3.5μm），柱温为40°C。两种方法均使用配备电喷雾离子源的Sciex Qtrap 5500质谱仪，在正离子模式下进行多反应监测。这两种方法在血清样本50.0 - 20000 ng/ml和尿液样本50.0 - 10000 ng/ml范围内均显示出良好的线性、准确性和精密度，回收率高且基质效应最小。未观察到残留效应。在两种方法的储存、制备和分析条件下，依替米星在人血清或尿液样本中保持稳定。这两种简单可靠的方法成功应用于依替米星在中国健康志愿者静脉注射单剂量和多剂量后的剂量递增I期临床试验。基于这两种方法，我们首次确定了依替米星在人体内的综合药代动力学，这将进一步用于探索折点研究。
