Han Dong-Gyun, Kwak Jinsook, Seo Seong-Wook, Kim Ji-Min, Yoo Jin-Wook, Jung Yunjin, Lee Yun-Hee, Kim Min-Soo, Jung Young-Suk, Yun Hwayoung, Yoon In-Soo
Department of Manufacturing Pharmacy, College of Pharmacy, Pusan National University, Busan 46241, Korea.
Department of Pharmacy, College of Pharmacy, Seoul National University, Seoul 08826, Korea.
Pharmaceutics. 2019 Aug 2;11(8):382. doi: 10.3390/pharmaceutics11080382.
Since diabetes mellitus and osteoarthritis are highly prevalent diseases, combinations of antidiabetic agents like repaglinide (REP) and non-steroidal anti-inflammatory drugs (NSAID) like celecoxib (CEL) could be commonly used in clinical practice. In this study, a simple and sensitive bioanalytical HPLC method combined with fluorescence detector (HPLC-FL) was developed and fully validated for simultaneous quantification of REP and CEL. A simple protein precipitation procedure and reversed C18 column with an isocratic mobile phase (mixture of ACN and pH 6.0 phosphate buffer) were employed for sample preparation and chromatographic separation. The fluorescence detector was set at a single excitation/emission wavelength pair of 240 nm/380 nm. The linearity (10-2000 ng/mL), accuracy, precision, extraction recovery, matrix effect, and stability for this method were validated as per the current FDA guidance. The bioanalytical method was applied to study pharmacokinetic interactions between REP and CEL in vivo, successfully showing that concurrent administration with oral REP significantly altered the pharmacokinetics of oral CEL. Furthermore, an in vitro metabolism and protein binding study using human materials highlighted the possibility of metabolism-based interactions between CEL and REP in clinical settings.
由于糖尿病和骨关节炎是高度流行的疾病,瑞格列奈(REP)等抗糖尿病药物与塞来昔布(CEL)等非甾体抗炎药的联合应用在临床实践中可能会经常使用。在本研究中,开发了一种简单灵敏的生物分析HPLC方法并结合荧光检测器(HPLC-FL),用于同时定量REP和CEL,并进行了全面验证。采用简单的蛋白沉淀程序和具有等度流动相(乙腈和pH 6.0磷酸盐缓冲液的混合物)的反相C18柱进行样品制备和色谱分离。荧光检测器设置在240 nm/380 nm的单一激发/发射波长对。根据当前FDA指南对该方法的线性(10 - 2000 ng/mL)、准确度、精密度、提取回收率、基质效应和稳定性进行了验证。该生物分析方法被应用于研究REP和CEL在体内的药代动力学相互作用,成功表明口服REP同时给药显著改变了口服CEL的药代动力学。此外,一项使用人体材料的体外代谢和蛋白结合研究突出了临床环境中CEL和REP之间基于代谢的相互作用的可能性。