CXCR4 targeting by TPCA‑1 attenuates neuroinflammation in intracerebral hemorrhage through inhibiting NF‑κB and AP‑1 activation.

Intracerebral hemorrhage (ICH)-induced secondary brain injury (SBI) is a critical contributor to poor neurological outcomes. In this study, we aimed to identify potential small‑molecule therapeutic agents and systematically validate their neuroprotective efficacy. Bioinformatics analysis was performed to screen ICH‑related hub genes, and Connectivity Map (CMap) was used to predict candidate compounds, followed by molecular docking to verify their binding affinity. The therapeutic effect of TPCA‑1 was further evaluated in a mouse ICH model using neurological scoring, cerebral edema assessment, qPCR, Western blot, and ELISA. Bioinformatics analysis indicated that the chemokine signaling pathway is critically involved in ICH pathogenesis. Molecular docking revealed that TPCA‑1 displayed the strongest binding affinity for CXCR4 among all predicted compounds. In vivo, 20 mg/kg TPCA‑1 significantly ameliorated neurological deficits and reduced cerebral edema in ICH mice. Mechanistically, TPCA-1 interacted with CXCR4, as supported by molecular docking, MST, and CETSA assays, and suppressed downstream inflammatory signaling involving NF-κB and AP-1, thereby decreasing the production of pro‑inflammatory cytokines including TNF‑α, IL‑1β, and IL‑6. Importantly, Co-treatment with the CXCR4 inhibitor AMD3100 did not produce additional inhibitory effects. Collectively, TPCA‑1 exerts neuroprotective effects against ICH‑induced SBI by targeting CXCR4, which further inhibits the activation of NF‑κB and AP‑1 and ultimately alleviates neuroinflammation.