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

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.