Unlocking the potential: unveiling tyrphostins with Michael-reactive cyanoacrylate motif as promising inhibitors of human 5-lipoxygenase.

IF 2.9 4区 医学 Q2 PHYSIOLOGY
Maximilian Molitor, Amelie Menge, Sebastian Mandel, Sven George, Susanne Müller, Stefan Knapp, Bettina Hofmann, Dieter Steinhilber, Ann-Kathrin Häfner
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Abstract

Human 5-lipoxygenase (5-LO) is the key enzyme in the biosynthesis of leukotrienes, mediators of the innate immune system that also play an important role in inflammatory diseases and cancer. In this study, we present compounds, containing a Michael-reactive cyanoacrylate moiety as potent inhibitors of 5-LO. Representatives of the tyrosine kinase inhibitor family called tyrphostins, structurally related to known 5-LO inhibitors, were screened for their 5-LO inhibitory properties using recombinant human 5-LO, intact human PMNL (polymorphonuclear leukocytes), and PMNL homogenates. Their mode of action was characterized by the addition of glutathione, using a fourfold cysteine 5-LO mutant and mass spectrometry analysis. SAR studies revealed several members of the tyrphostin family containing a Michael-reactive cyanoacrylate to efficiently inhibit 5-LO. We identified degrasyn (IC50 0.11 µM), tyrphostin A9 (IC50 0.8 µM), AG879 (IC50 78 nM), and AG556 (IC50 64 nM) as potent 5-LO inhibitors. Mass spectrometry analysis revealed that degrasyn and AG556 covalently bound to up to four cysteines, including C416 and/or C418 which surround the substrate entry site. Furthermore, the 5-LO inhibitory effect of degrasyn was remarkably impaired by the addition of glutathione or by the mutation of cysteines to serines at the surface of 5-LO. We successfully identified several tyrphostins as potent inhibitors of human 5-LO. Degrasyn and AG556 were able to covalently bind to 5-LO via their cyanoacrylate moiety. This provides a promising mechanism for targeting 5-LO by Michael acceptors, leading to new therapeutic opportunities in the field of inflammation and cancer.

挖掘潜力:揭示具有迈克尔反应氰基丙烯酸酯基团的酪肽类药物作为人类 5-脂氧合酶抑制剂的前景。
人类 5-脂氧合酶(5-LO)是生物合成白三烯的关键酶,白三烯是先天性免疫系统的介质,在炎症性疾病和癌症中也发挥着重要作用。在这项研究中,我们发现了含有迈克尔反应氰基丙烯酸酯分子的化合物,它们是 5-LO 的强效抑制剂。研究人员使用重组人 5-LO、完整的人 PMNL(多形核白细胞)和 PMNL 均质物筛选了酪氨酸激酶抑制剂家族中与已知的 5-LO 抑制剂结构相关的代表化合物--tyrphostins。通过添加谷胱甘肽、使用四倍半胱氨酸 5-LO 突变体和质谱分析,确定了它们的作用模式。SAR研究显示,含有迈克尔反应氰基丙烯酸酯的tyrphostin家族的几个成员能有效抑制5-LO。我们发现degrasyn(IC50 0.11 µM)、tyrphostin A9(IC50 0.8 µM)、AG879(IC50 78 nM)和AG556(IC50 64 nM)是有效的5-LO抑制剂。质谱分析表明,degrasyn 和 AG556 与多达四个半胱氨酸共价结合,包括围绕底物进入位点的 C416 和/或 C418。此外,加入谷胱甘肽或将 5-LO 表面的半胱氨酸突变为丝氨酸都会明显削弱 degrasyn 对 5-LO 的抑制作用。我们成功鉴定了几种酪磷脂类药物作为人类 5-LO 的强效抑制剂。Degrasyn 和 AG556 能够通过其氰基丙烯酸酯分子与 5-LO 共价结合。这为迈克尔受体靶向 5-LO 提供了一种很有前景的机制,从而为炎症和癌症领域带来了新的治疗机会。
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来源期刊
CiteScore
8.80
自引率
2.20%
发文量
121
审稿时长
4-8 weeks
期刊介绍: Pflügers Archiv European Journal of Physiology publishes those results of original research that are seen as advancing the physiological sciences, especially those providing mechanistic insights into physiological functions at the molecular and cellular level, and clearly conveying a physiological message. Submissions are encouraged that deal with the evaluation of molecular and cellular mechanisms of disease, ideally resulting in translational research. Purely descriptive papers covering applied physiology or clinical papers will be excluded. Papers on methodological topics will be considered if they contribute to the development of novel tools for further investigation of (patho)physiological mechanisms.
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