胰岛素调节的氨基肽酶的新变构抑制剂的发现为酶的机制提供了新的见解。

IF 4.3 3区 化学 Q2 CHEMISTRY, MULTIDISCIPLINARY
ACS Omega Pub Date : 2025-04-23 eCollection Date: 2025-05-06 DOI:10.1021/acsomega.5c01169
Galateia Georgaki, Nikoletta-Maria Koutroumpa, Panagiotis Lagarias, Antreas Afantitis, Athanasios Papakyriakou, Efstratios Stratikos
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引用次数: 0

摘要

胰岛素调节氨基肽酶(IRAP)是一种跨膜锌金属蛋白酶,具有多种重要的生物学作用,包括纤维化、脓毒性血栓形成、认知功能和免疫系统调节。因此,IRAP是几种疾病的新兴药理靶点。然而,由于其高序列和与许多其他具有高度保守活性位点的酶的功能同源性,开发特异性调节其活性的选择性抑制剂仍然具有挑战性。为了规避这一限制,我们瞄准了苹果酸酯变构位点,这是一个产生高选择性同源酶ERAP1抑制剂的位点。我们进行了虚拟筛选,以发现与IRAP中该变构位点具有高亲和力的药物样化合物。一个包含3800万种来自ENAMINE的药物样化合物的数据库被用于筛选目标位点的三种构象。随后,通过分子动力学模拟和比较MM/GBSA自由能计算对排名靠前的化合物进行评估,从中选择17个化合物,通过两个正交试验进一步评估其体外抑制IRAP的活性。选择三个命中点(每个酶构象和底物类别各一个)进行进一步的机制评估,揭示底物依赖的非竞争性或非竞争性作用机制,与变构位点的构象敏感性一致。我们的研究结果支持苹果酸位点的可追踪性,发现了新的选择性IRAP抑制剂,为进一步开发建立了新的靶点,并表明可能通过变构抑制剂靶向IRAP酶的不同构象来靶向IRAP的特定生物学功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Discovery of Novel Allosteric Inhibitor Hits for Insulin-Regulated Aminopeptidase Provides Insights on Enzymatic Mechanism.

Insulin-regulated aminopeptidase (IRAP) is a transmembrane zinc metalloprotease with various important biological roles, including fibrosis, septic thrombosis, cognitive functions, and immune system regulation. As a result, IRAP is an emerging pharmacological target for several diseases. However, the development of selective inhibitors that specifically regulate its activity remains challenging due to its high sequence and functional homology with many other enzymes that have highly conserved active sites. To circumvent this limitation, we targeted the malate allosteric site, a site that has yielded highly selective inhibitors of the homologous enzyme ERAP1. We performed virtual screening to discover drug-like compounds that bind with high affinity to this allosteric site in IRAP. A database of 38 million diverse, drug-like compounds from ENAMINE was employed for screening at three conformations of the targeted site. A subset of the top-ranked compounds was subsequently evaluated using molecular dynamics simulations and comparative MM/GBSA free energy calculation, from which 17 were selected for further in vitro evaluation of their inhibitory activity for IRAP by two orthogonal assays. Three hits, one for each enzyme conformation and substrate class, were selected for further mechanistic evaluation revealing substrate-dependent uncompetitive or noncompetitive mechanisms of action, consistent with the conformationally sensitive nature of the allosteric site. Our results support the tractability of the malate site for the discovery of novel selective IRAP inhibitors, establish novel hits for further development, and suggest that it may be possible to target specific biological functions of IRAP by targeting distinct conformations of the enzyme by allosteric inhibitors.

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来源期刊
ACS Omega
ACS Omega Chemical Engineering-General Chemical Engineering
CiteScore
6.60
自引率
4.90%
发文量
3945
审稿时长
2.4 months
期刊介绍: ACS Omega is an open-access global publication for scientific articles that describe new findings in chemistry and interfacing areas of science, without any perceived evaluation of immediate impact.
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