A novel molecule ZYZ311 derivatized from Carvacrol ameliorates DSS-induced colitis in mice via inhibiting JAK2/STAT3.

Abstract

BACKGROUND: Carvacrol, a monoterpenoid phenol, exhibits notable anti-inflammatory activity but is limited by its poor bioavailability and low target specificity. To overcome these drawbacks, we designed and synthesized a novel hydrazone derivative, ZYZ311, to enhance its pharmacological properties. METHODS: The anti-inflammatory effects of ZYZ311 were evaluated in LPS-stimulated Raw264.7 macrophages and a dextran sulfate sodium (DSS)-induced mouse model of acute ulcerative colitis. Inflammatory factors were quantified by ELISA, and gene/protein expression levels were analyzed via RT-PCR immunohistochemistry, immunofluorescence, and Western blot. The mechanism of action of ZYZ311 in Raw264.7 macrophages was investigated using RNA sequencing analysis. Direct compound-target interactions were assessed using molecular docking and cellular thermal shift assays. RESULTS: ZYZ311 significantly inhibited nitric oxide (NO) production and suppressed IL-6, COX-2, and NOS2 expression in RAW264.7 macrophages. Transcriptomic analysis showed that ZYZ311 selectively inhibited the levels of different inflammatory factors. Mechanistically, ZYZ311 acted predominantly during the mid-to-late phase of inflammation by directly binding to STAT3 and inhibiting its phosphorylation at Tyr705, thereby blocking IL-6-induced JAK2-STAT3 signaling. Notably, ZYZ311 had minimal effect on early NF-κB activation. In vivo, ZYZ311 effectively alleviated the clinical symptoms of acute colitis, reduced histopathological damage, and decreased STAT3 activation in the colonic tissue. CONCLUSIONS: Our results demonstrate that hydrazone modification of carvacrol (resulting in ZYZ311) significantly improves its anti-inflammatory potency and target selectivity. ZYZ311 serves as a promising lead compound for the development of natural product-derived therapeutics.