Giri Poonam, Gupta Lakshmikant, Rathod Anil, Joshi Vipul, Giri Shyamkumar, Patel Nirmal, Agarwal Sameer, R Jain Mukul
Department of Drug Metabolism and Pharmacokinetics, Zydus Research Centre, Moraiya, Ahmadabad, Gujarat, India.
Department of Medicinal Chemistry, Zydus Research Centre, Moraiya, Ahmadabad, Gujarat, India.
Drug Metab Lett. 2022 Mar 15. doi: 10.2174/1872312815666220315145945.
Identification of clinical drug-drug interaction (DDI) risk is an important aspect of drug discovery and development owing to poly-pharmacy in present-day clinical therapy. Drug metabolizing enzymes (DME) plays important role in the efficacy and safety of drug candidates. Hence evaluation of a New Chemical Entity (NCE) as a victim or perpetrator is very crucial for DDI risk mitigation. ZY12201 (2-((2-(4-(1H-imidazol-1-yl) phenoxy) ethyl) thio)-5-(2-(3, 4- dimethoxy phenyl) propane-2-yl)-1-(4-fluorophenyl)-1H-imidazole) is a novel and potent Takeda-G-protein-receptor-5 (TGR-5) agonist. ZY12201 was evaluated in-vitro to investigate the DDI liabilities.
The key objective was to evaluate the CYP inhibition potential of ZY12201 for an opportunity to use it as a tool compound for pan CYP inhibition activities.
In-vitro drug metabolizing enzymes (DME) inhibition potential of ZY12201 was evaluated against major CYP isoforms (1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4/5), aldehyde oxidase (AO), monoamine oxidase (MAO), and flavin-containing monooxygenase (FMO in human liver cytosol/mitochondrial preparation/ microsomes using probe substrates and Liquid Chromatography with tandem mass spectrometry (LC-MS-MS) method.
The study conducted on ZY12201 at 100 µM ZY12201 was found to reduce the metabolism of vanillin (AO probe substrate), tryptamine (MAO probe substrate), and benzydamine (FMO probe substrate) by 49.2%, 14.7%, and 34.9%, respectively. ZY12201 Ki values were 0.38, 0.25, 0.07, 0.01, 0.06, 0.02, 7.13, 0.03 and 0.003 μM for CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4/5 (substrate: testosterone) and CYP3A4/5 (substrate: midazolam), respectively. Time-dependant CYP inhibition potential of ZY12201 was assessed against CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4/5 and no apparent IC50 shift was observed.
ZY12201, at 100 µM concentration showed low inhibition potential of AO, MAO, and FMO. ZY12201 was found as a potent inhibitor of CYP1A2, 2B6, 2C8, 2C9, 2C19, 2D6, and 3A4/5 while moderately inhibits to CYP2E1. Inhibition of CYP1A2, CYP2B6, CYP2C19, and CYP2E1 by ZY12201 was competitive, while inhibition of CYP2C8, CYP2C9, CYP2D6, and CYP3A4/5 was of mixed-mode. ZY12201 is a non-time-dependent inhibitor of CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4/5. In summary, the reported Ki values unequivocally support that ZY12201 has a high potential to inhibit all major CYP isoforms. ZY12201 can be effectively used as a tool compound for in-vitro evaluation of CYP-based metabolic contribution to total drug clearance in the lead optimization stage of Drug Discovery Research.
由于当今临床治疗中存在多种药物联用的情况，识别临床药物 - 药物相互作用（DDI）风险是药物发现和开发的一个重要方面。药物代谢酶（DME）在候选药物的疗效和安全性方面发挥着重要作用。因此，评估新化学实体（NCE）作为受害者或肇事者对于减轻DDI风险非常关键。ZY12201（2 - ((2 - (4 - (1H - 咪唑 - 1 - 基)苯氧基)乙基)硫代)-5 - (2 - (3,4 - 二甲氧基苯基)丙烷 - 2 - 基)-1 - (4 - 氟苯基)-1H - 咪唑）是一种新型且强效的武田 - G - 蛋白受体 - 5（TGR - 5）激动剂。对ZY12201进行了体外研究以调查其DDI倾向。
关键目标是评估ZY12201对细胞色素P450（CYP）的抑制潜力，以便有机会将其用作泛CYP抑制活性的工具化合物。
使用探针底物以及液相色谱 - 串联质谱（LC - MS - MS）方法，在人肝细胞溶胶/线粒体制剂/微粒体中评估ZY12201对主要CYP同工酶（1A2、2B6、2C8、2C9、2C19、2D6、2E1和3A4/5）、醛氧化酶（AO）、单胺氧化酶（MAO）和含黄素单加氧酶（FMO）的体外药物代谢酶（DME）抑制潜力。
在100 μM ZY12201浓度下对ZY12201进行的研究发现，它分别使香草醛（AO探针底物）、色胺（MAO探针底物）和苄达明（FMO探针底物）的代谢降低了49.2%、14.7%和34.9%。ZY12201对CYP1A2、CYP2B6、CYP2C8、CYP2C9、CYP2C19、CYP2D6、CYP2E1、CYP3A4/5（底物：睾酮）和CYP3A4/5（底物：咪达唑仑）的抑制常数（Ki）值分别为0.38、0.25、0.07、0.01、0.06、0.02、7.13、0.03和0.003 μM。针对CYP1A2、CYP2B6、CYP2C8、CYP2C9、CYP2C,19、CYP2D6、CYP2E1和CYP3A4/5评估了ZY12201的时间依赖性CYP抑制潜力，未观察到明显的半数抑制浓度（IC50）偏移。
在100 μM浓度下，ZY12201对AO、MAO和FMO显示出较低的抑制潜力。发现ZY12201是CYP1A2、2B6、2C8、2C9、2C19、2D6和3A4/5的强效抑制剂，而对CYP2E1有中度抑制作用。ZY12201对CYP1A2、CYP2B6、CYP2C19和CYP2E1的抑制作用是竞争性的，而对CYP2C8、CYP2C9、CYP2D6和CYP3A4/5的抑制作用是混合模式。ZY12201是CYP1A2、CYP,2B6、CYP2C8、CYP2C9、CYP2C19、CYP2D6、CYP2E1、CYP3A4/5的非时间依赖性抑制剂。总之，所报道的Ki值明确支持ZY12201具有抑制所有主要CYP同工酶的高潜力。在药物发现研究的先导优化阶段，ZY,12201可有效地用作工具化合物，用于体外评估基于CYP的代谢对总药物清除率的贡献。
