Discovery of a Novel Series of <i>Homo sapiens</i> Caseinolytic Protease P Agonists for Colorectal Adenocarcinoma Treatment via ATF3-Dependent Integrated Stress Response

Abstract

<i>Homo sapiens</i> caseinolytic protease P (HsClpP) activation is a promising strategy for colon cancer treatment. In this study, <b>CCG1423</b> was identified as a selective activator of HsClpP. After optimization, <b>NCA029</b> emerged as the most potent compound, with an EC₅₀ of 0.2 μM against HsClpP. Molecular dynamics revealed that the affinity of <b>NCA029</b> for the YYW aromatic network is crucial for its selectivity toward HsClpP. Furthermore, <b>NCA029</b> displayed favorable pharmacokinetics and safety profiles and significantly inhibited tumor growth in HCT116 xenografts, resulting in 83.6% tumor inhibition. Mechanistically, <b>NCA029</b> targeted HsClpP, inducing mitochondrial dysfunction and activating the ATF3-dependent integrated stress response, ultimately causing cell death in colorectal adenocarcinoma. These findings highlight <b>NCA029</b> as an effective HsClpP activator with potential for colon cancer therapy.