许多间变性淋巴瘤激酶（ALK）抑制剂已获得临床批准，其中劳拉替尼（lorlatinib），特别是作为第三代药物，显示出针对各种耐药 ALK 单突变的功效。然而，劳拉替尼的持续临床使用导致出现了赋予劳拉替尼耐药性的 ALK 双突变，特别是 ALKL1196M/G1202R。 TPX-0131是目前正在开发的潜在第四代ALK抑制剂。 TPX-0131 表现出更广泛的抗 ALK 耐药突变活性，可有效抑制 26 种单点突变和各种双/三重突变，包括溶剂前沿突变和看门人突变。本研究首次通过建模、MD模拟、自由能计算和US模拟，全面阐明了TPX-0131克服lorlatinib对ALKL1196M/G1202R耐药的分子机制。结果表明，劳拉替尼与铰链区关键残基之间的相互作用受到L1196M/G1202R双突变的干扰，导致Glu1197与劳拉替尼之间重要氢键的破坏。对于 TPX-0131，L1196M/G1202R 突变增强了静电和范德华相互作用，主要在铰链区、G 环和 β 链中引起显着的构象变化。 TPX-0131 与残基 Arg1202、Met1199 和 Arg1120 的紧密结合有助于克服 ALKL1196M/G1202R 突变体中的洛拉替尼耐药性。这些研究结果预计将有助于深入了解 TPX-0131 治疗 ALKG1202R/L1196M 诱导的 NSCLC 耐药的机制以及优化 ALK 抑制剂。版权所有 © 2024 Elsevier Ltd。保留所有权利。

A number of anaplastic lymphoma kinase (ALK) inhibitors have been clinically approved, with lorlatinib, particularly as a third-generation drug, demonstrating efficacy against various drug-resistant ALK single mutations. However, continued clinical use of lorlatinib has led to the emergence of ALK double mutations conferring resistance to lorlatinib, notably ALKL1196M/G1202R. TPX-0131 is a potential fourth-generation ALK inhibitor currently under development. TPX-0131 demonstrates a broader spectrum of activity against ALK-resistant mutations, efficiently inhibiting 26 single-point mutations and various double/triple mutations, including solvent front mutations and gatekeeper mutations. In this study, for the first time, a comprehensive elucidation of the molecular mechanisms by which TPX-0131 overcomes lorlatinib resistance to ALKL1196M/G1202R through modeling, MD simulations, free energy calculations, and US simulations. The results indicate that the interactions between lorlatinib and key residues at the hinge region are disturbed by L1196M/G1202R double mutation, leading to the disruption of important hydrogen bonding between Glu1197 and lorlatinib. For TPX-0131, the L1196M/G1202R mutation enhances electrostatic and van der Waals interactions, causing significant conformational changes primarily in the hinge region, G-loop, and β-strands. The tight binding of TPX-0131 to residues Arg1202, Met1199 and Arg1120 contribute significantly to overcoming lorlatinib resistance in ALKL1196M/G1202R mutant. These research results are expected to offer insights into the mechanism of TPX-0131 in treating ALKG1202R/L1196M-induced NSCLC resistance and optimizing of ALK inhibitors.Copyright © 2024 Elsevier Ltd. All rights reserved.