Targeting NLRP3 is a highly promising strategy for treating uncontrolled inflammation, which can cause a wide range of diseases or promote disease progression. More NLRP3-targeting inhibitors with different scaffolds are needed to increase the chances of developing safe and effective NLRP3 inhibitors and treating inflammation in different tissues. Here, we discovered the novel quinoline analogues that exhibit potent inhibitory activity against the NLRP3/IL-1beta pathway in J774A.1, BMDMs, and human peripheral blood cells. Mechanistic studies confirmed W16 may directly target NLRP3 and block the NLRP3 inflammasome assembly and activation. In vitro studies demonstrated that W16 has potent anti-inflammatory effects on DSS-induced ulcerative colitis model. Our findings demonstrated that W16 is a potential lead compound targeting NLRP3 and deserves further investigation for the treatment of NLRP3-related inflammatory diseases.