Kuschak Markus, Schlegel Jonathan G, Schneider Marion, Kehraus Stefan, Voss Jan H, Seidinger Alexander, Matthey Michaela, Wenzel Daniela, Fleischmann Bernd K, König Gabriele M, Müller Christa E
PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical & Medicinal Chemistry, University of Bonn, Bonn, Germany.
Institute of Pharmaceutical Biology, University of Bonn, Bonn, Germany.
Front Chem. 2020 Sep 24;8:833. doi: 10.3389/fchem.2020.00833. eCollection 2020.
The cyclic depsipeptide FR900359 (FR) isolated from the plant and produced by endosymbiotic bacteria acts as a selective Gq protein inhibitor. It is a powerful tool to study G protein-coupled receptor signaling, and has potential as a novel drug for the treatment of pulmonary diseases and cancer. For pharmacokinetic studies, sensitive quantitative measurements of drug levels are required. In the present study we established an LC-MS/MS method to detect nanomolar concentrations of FR and the structurally related natural product YM-254890 (YM) in biological samples. HPLC separation coupled to ESI-QTOF-MS and UV-VIS detection was applied. For identification and quantification, the extract ion chromatogram (EIC) of M+1 was evaluated. Limits of detection (LOD) of 0.53-0.55 nM and limits of quantification (LOQ) of 1.6-1.7 nM were achieved for both FR and YM. This protocol was subsequently applied to determine FR concentrations in mouse organs and tissues after peroral application of the drug. A three-step liquid-liquid extraction protocol was established, which resulted in adequate recovery rates of typically around 50%. The results indicated low peroral absorption of FR. Besides the gut, highest concentrations were determined in eye and kidney. The developed analytical method will be useful for preclinical studies to evaluate these potent Gq protein inhibitors, which may have potential as future drugs for complex diseases.
从植物中分离并由内共生细菌产生的环缩肽FR900359（FR）作为一种选择性Gq蛋白抑制剂。它是研究G蛋白偶联受体信号传导的有力工具，并且具有作为治疗肺部疾病和癌症的新型药物的潜力。对于药代动力学研究，需要对药物水平进行灵敏的定量测量。在本研究中，我们建立了一种液相色谱-串联质谱法，用于检测生物样品中纳摩尔浓度的FR以及结构相关的天然产物YM-254890（YM）。采用了与电喷雾-四极杆飞行时间质谱联用的高效液相色谱分离以及紫外-可见检测。为了进行鉴定和定量，评估了M+1的提取离子色谱图（EIC）。FR和YM的检测限（LOD）均达到0.53 - 0.55 nM，定量限（LOQ）均达到1.6 - 1.7 nM。该方法随后被应用于测定口服给药后小鼠器官和组织中的FR浓度。建立了三步液-液萃取方法，其回收率通常约为50%，结果表明FR的口服吸收率较低。除肠道外，在眼睛和肾脏中检测到的浓度最高。所建立的分析方法将有助于临床前研究评估这些有效的Gq蛋白抑制剂，它们可能具有作为未来治疗复杂疾病药物的潜力。
