National Institute of Pharmaceutical Education and Research-Ahmedabad (NIPER-A), An Institute of National Importance, Government of India, Department of Pharmaceuticals, Ministry of Chemicals and Fertilizers, Opp. Airforce Station, Palaj, Gandhinagar 382355, Gujarat, India.
National Institute of Pharmaceutical Education and Research-Ahmedabad (NIPER-A), An Institute of National Importance, Government of India, Department of Pharmaceuticals, Ministry of Chemicals and Fertilizers, Opp. Airforce Station, Palaj, Gandhinagar 382355, Gujarat, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Nov 15;1185:123011. doi: 10.1016/j.jchromb.2021.123011. Epub 2021 Oct 28.
A systematic metabolite profiling approach has paramount importance in detecting, identifying, and characterizing drug metabolites. Till date, there is no report published on the comprehensive metabolic fate of flibanserin (FLB). In this study, the structure of entire potential metabolites of FLB has been elucidated by execution of in silico tool and high resolution mass spectrometry based metabolite profiling strategy employing data-dependent and data-independent approaches. In vitro metabolism profile was investigated after incubating FLB with liver microsomes (rat and human) and S9 fractions in presence of their respective co-factors. In vivo metabolites were identified from rat plasma, urine, feces, and brain tissue samples. An efficient extraction technique was developed that made it possible to identify the metabolites generated even in extremely low concentrations. Extraction was carried out by precipitating protein and thereafter solid-phase extraction to enrich their concentration in the sample before analysis. Fourteen new metabolites have been identified and characterized. Most of the metabolites of FLB were generated due to hydrolysis and oxidation followed by glucuronide, sulfate, and methyl conjugation. Additionally, a spiking study was employed to confirm the presence of N-oxide metabolite in human liver S9 fraction and rat urine samples. Moreover, we have established the probable biotransformation pathway of FLB and successfully analyzed the toxicity potential of the metabolites using Pro Tox-II software.
系统的代谢物分析方法在检测、鉴定和描述药物代谢物方面具有重要意义。迄今为止，尚无关于氟班色林（FLB）全面代谢命运的报道。在这项研究中，通过执行基于计算机的工具和基于高分辨率质谱的代谢物分析策略，采用数据依赖和数据独立的方法，阐明了 FLB 的整个潜在代谢物的结构。在存在各自辅助因子的情况下，用大鼠和人肝微粒体和 S9 级分孵育 FLB 后，研究了体外代谢谱。从大鼠血浆、尿液、粪便和脑组织样本中鉴定了体内代谢物。开发了一种有效的提取技术，使得即使在极低浓度下产生的代谢物也能够被识别。通过沉淀蛋白质并进行固相萃取，在分析前浓缩样品中的代谢物，从而进行提取。已经鉴定和表征了 14 种新的代谢物。FLB 的大多数代谢物是由于水解和氧化，然后是葡萄糖醛酸、硫酸盐和甲基化缀合而产生的。此外，还进行了加标研究以确认人肝 S9 级分和大鼠尿液样品中存在 N-氧化物代谢物。此外，我们已经建立了 FLB 的可能生物转化途径，并成功使用 Pro Tox-II 软件分析了代谢物的毒性潜力。
