Investigation of novel ATX inhibitor metabolites by UHPLC-orbitrap-MS/MS and molecular docking studies
Investigation of novel ATX inhibitor metabolites by UHPLC-orbitrap-MS/MS and molecular docking studies
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通过 UHPLC-orbitrap-MS/MS 和分子对接研究研究新型 ATX 抑制剂代谢物
DOI：
10.1016/j.jpba.2022.114606
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发表时间：
2022
影响因子：
3.4
通讯作者：
Lixin Sun
中科院分区：
文献类型：
--
作者：
Hongjin Wang;Yuxiang Chen;Xinjie Qi;Hao Zhang;Xin Zhai;Lixin Sun
溶血磷脂酸（LPA）介导的信号传导与肺纤维化和肺细胞增殖有关，例如促进成纤维细胞积聚和刺激TGF-β信号。减少溶血磷脂酸产生的基本机制之一是抑制自分泌运动因子活性，其可以阻断溶血磷脂酰胆碱（LPC）水解成LPA，从而导致肺纤维化发展的缓解。近年来，自分泌运动因子抑制剂3，4-二氟苄基（1-乙基-5-（4-（（4-羟基哌啶-1-基）-甲基）-噻唑-2-基）-1H-吲哚-3-基）氨基甲酸酯（NAI 59）表现出显著的抗肺纤维化作用，但NAI 59的代谢信息尚未得到全面阐述。因此，在本研究中，我们的目的是确定NAI 59在大鼠体内的代谢产物，使用超高效液相色谱-高分辨率串联质谱，我们分析了在血浆中的代谢产物的潜在抗纤维化与晶体结构autotaxin分子对接。结果共鉴定出24种代谢产物，即15种I相代谢产物和9种II相代谢产物，均为新发现的代谢产物。NAI 59的代谢途径主要是羟基化、氧化、氢化、葡萄糖醛酸结合、硫酸盐结合、乙酰化和葡萄糖基化。分子对接结果表明，血浆中的代谢产物与autotaxin的活性位点有很强的对接亲和力，可能具有潜在的抗肺纤维化活性。相互作用图揭示了氟苯基和哌啶上的羟基的重要性，这些羟基有助于与自分泌运动因子形成氢键。此外，我们发现，当结构中含有葡萄糖醛酸基团时，代谢产物具有更好的对接结果。据我们所知，这项研究是第一次研究NAI 59在大鼠中的代谢命运，我们使用分子对接分析了代谢产物的潜在抗纤维化方面。这项研究为进一步的临床前研究提供了代谢理论基础，并为开发改进的ATX抑制剂提供了新的见解。
Lysophosphatidic acid (LPA) mediated signaling is relevant to fibrosis and the proliferation of pulmonary cells, such as the promotion of fibroblast accumulation and stimulation of the TGF-β signal. One of the essential mechanisms to reduce lysophosphatidic acid production is inhibiting autotaxin activity, which can block the hydrolysis of a lysophosphatidylcholine (LPC) into LPA, leading to the alleviation of the development of pulmonary fibrosis. Recently, 3,4-difluorobenzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)-methyl)-thiazol-2-yl)-1H-indol-3-yl)carbamate (NAI59), a novel autotaxin inhibitor, showed outstanding anti-pulmonary fibrosis therapeutic effectiveness; however, the metabolic information of NAI59 has not been comprehensively expounded yet. Therefore, in this study, we aim to identify the metabolites of NAI59 in rats using ultra-high-performance liquid chromatography-high-resolution tandem mass spectrometry, and we analyze the potential anti-fibrosis of metabolites in plasma using molecular docking with the crystal structure autotaxin. As a result, a total of 24 metabolites, namely, 15 phase I metabolites and 9 phase II metabolites, were identified, all of which are the novel discovered metabolites. The metabolic pathways of NAI59 were primarily hydroxylation, oxidation, hydrogenation, glucuronic acid conjugation, sulfate conjugation, acetylation, and glucosylation. The molecular docking result showed that the identified metabolites in plasma had great affinities upon docking into the active site of autotaxin, indicating they may have potential anti-pulmonary fibrosis activity. The interaction plots revealed the importance of fluorophenyl and the hydroxyl groups on piperidine that contribute toward the formation of hydrogen bonds with autotaxin. Additionally, we found that the metabolite possessed a better docking result when the structure contained a glucuronic acid group. To the best of our knowledge, this study is the first research into the metabolic fate of NAI59 in rats, and we analyze the potential anti-fibrosis aspects of metabolites using molecular docking. This study provides a metabolic rationale for further preclinical research and imparts new insights for developing improved ATX inhibitors.