复制起点组装是染色体DNA复制的关键步骤。在此过程中，ORC 复合物结合 DNA，并与 CDC6 和 CDT1 一起促进 MCM 解旋酶的加载。针对起始组装的化学物质可能有助于使高度增殖的癌细胞变得敏感。然而，由于该过程的多阶段性质，识别此类化合物具有挑战性。在这里，我们使用非洲爪蟾卵提取物建立了高通量筛选来分离 MCM 染色质负载抑制剂，从而确定 NSC-95397 是一种强大的复制起点组装抑制剂，可靶向 CDC6 蛋白并促进其降解。使用开发来测试选择性药物诱导致死率的系统，我们发现 NSC-95397 会触发具有较高增殖能力的人类细胞和非洲爪蟾胚胎的细胞死亡。这些发现证明了破坏 DNA 复制过程的分子在靶向过度增殖细胞方面的有效性，突出了它们作为抗癌分子的潜力。© 2024 作者。

Replication origin assembly is a pivotal step in chromosomal DNA replication. In this process, the ORC complex binds DNA and, together with the CDC6 and CDT1, promotes the loading of the MCM helicase. Chemicals targeting origin assembly might be useful to sensitize highly proliferative cancer cells. However, identifying such compounds is challenging due to the multistage nature of this process. Here, using Xenopus laevis egg extract we set up a high-throughput screening to isolate MCM chromatin loading inhibitors, which led to the identification of NSC-95397 as a powerful inhibitor of replication origin assembly that targets CDC6 protein and promotes its degradation. Using systems developed to test selective drug-induced lethality we show that NSC-95397 triggers cell death both in human cells and Xenopus embryos that have higher proliferative ability. These findings demonstrate the effectiveness of molecules disrupting DNA replication processes in targeting hyperproliferating cells, highlighting their potential as anti-cancer molecules.© 2024 The Authors.