Inhibition of nuclear ROCK2 by a novel thiourea derivative induces potent antitumor effects through PTEN/FOXO1 pathway restoration in prostate cancer

Aberrant activation of the PI3K/Akt signaling pathway and frequent silencing of the tumor suppressor PTEN are central drivers of prostate cancer progression and therapeutic resistance. Addressing this dual dysregulation, we identified compound 25, a novel thiourea derivative, as a potent and selective anticancer agent. Among structurally related analogs, 25 exhibited the strongest antiproliferative activity in prostate cancer cells, while showing minimal cytotoxicity toward normal colon and kidney cells, indicating high selectivity for cancer cells. In vivo, 25 significantly suppressed tumor growth in a prostate cancer xenograft model in a dose-dependent manner, with greater efficacy than docetaxel. Target identification using AI-based target prediction platforms and kinase profiling revealed ROCK2 as the primary molecular target of 25. Mechanistically, 25 inhibited ROCK2 activity and nuclear expression, disrupting its interaction with transcriptional coactivators p300 and PGC-1α, thereby repressing oncogenic gene transcription, while restoring PTEN expression through the activation of FOXO1. This activation of PTEN led to suppression of PI3K/Akt signaling, cell cycle arrest, and apoptosis. Collectively, these findings uncover a nuclear transcriptional role of ROCK2 in sustaining oncogenic signaling and demonstrate that its inhibition by 25 restores tumor-suppressive PTEN/FOXO1 pathways. Compound 25 thus emerges as a promising therapeutic candidate for advanced prostate cancer.