微管靶向药物是最成功的抗癌药物之一。然而，耐药性的发展和不良反应的出现阻碍了它们的临床实施。迫切需要没有这种限制的新型微管靶向剂。通过采用基于基因表达的药物重新定位策略，本研究鉴定出 VU-0365114（最初是作为人毒蕈碱乙酰胆碱受体 M5 (M5 mAChR) 的正变构调节剂合成的）通过破坏微管的稳定性而成为一种新型的微管蛋白抑制剂。 VU-0365114 表现出广谱体外抗癌活性，特别是在结直肠癌细胞中。裸鼠肿瘤异种移植研究表明，VU-0365114 减缓了体内结直肠肿瘤的生长。 VU-0365114 的抗癌活性与其原始靶点 M5 mAChR 无关。此外，VU-0365114 不作为多药耐药 (MDR) 蛋白的底物，因此它可以克服 MDR。此外，激酶组分析显示 VU-0365114 没有表现出其他显着的脱靶效应。综上所述，我们的研究表明 VU-0365114 主要针对微管，尽管在进一步药物开发之前还需要进行更多研究，但为癌症治疗提供了重新利用的潜力。© 2023 作者。约翰·威利出版的《分子肿瘤学》

Microtubule-targeting agents represent one of the most successful classes of anticancer agents. However, the development of drug resistance and the appearance of adverse effects hamper their clinical implementation. Novel microtubule-targeting agents without such limitations are urgently needed. By employing a gene expression-based drug repositioning strategy, this study identifies VU-0365114, originally synthesized as a positive allosteric modulator of human muscarinic acetylcholine receptor M5 (M5 mAChR), as a novel type of tubulin inhibitor by destabilizing microtubules. VU-0365114 exhibits a broad-spectrum in vitro anticancer activity, especially in colorectal cancer cells. A tumor xenograft study in nude mice shows that VU-0365114 slowed the in vivo colorectal tumor growth. The anticancer activity of VU-0365114 is not related to its original target, M5 mAChR. In addition, VU-0365114 does not serve as a substrate of multidrug resistance (MDR) proteins, and thus, it can overcome MDR. Furthermore, a kinome analysis shows that VU-0365114 did not exhibit other significant off-target effects. Taken together, our study suggests that VU-0365114 primarily targets microtubules, offering potential for repurposing in cancer treatment, although more studies are needed before further drug development.© 2023 The Authors. Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.