Clofazimine can suppress tumor growth both in vitro and in vivo, making it a potential anticancer candidate. However, until now, the molecular mechanism of clofazimine in inhibiting cancer is not fully known. This study aims to analyze the possible mechanism of clofazimine in attenuating colitis-associated colon cancer (CAC) observed in the wnt/β-catenin signaling pathway. This study used male Balb/c (n = 36) mice which were randomly divided into six groups: normal control, negative control, curative group [dose of 0.2, 0.4, and 0.8 mg/20 g body weight (BW), respectively], and preventive group dose of 0.4 mg/day 20 g BB. Azoxymethane and dextran sodium sulfate (AOM/DSS) were used to induce colitis-related colon cancer. The enzyme-linked immunosorbent assay (ELISA) approach was used to assess the expression levels of caspase-3 and IL-1. H&E staining was used for histological investigation, followed by immunohistochemistry for catenin and axin-2 expression. The ELISA analysis showed that the curative dose of clofazimine 0.8 mg/20 g BW had lower IL-1β, β-catenin, axin-2, and caspase-3 expression compared to the AOM/DSS group. Histological analysis showed that clofazimine attenuates the inflammatory score compared to the AOM/DSS-only group. The best damage score reduction occurred in the curative dose group of 0.8 mg/kg BW and the preventive dose of 0.4 mg/20 g BW. The protein expression of β-catenin and axin-2 showed that the AOM/DSS-induced group had higher expression than the normal group. Decreased expression of both proteins was seen in a curative dose of clofazimine 0.8 mg/kg BW. Our study indicated that clofazimine has the potential to inhibit the growth of CAC in part through attenuation of the β-catenin signaling pathway, which includes attenuation of IL-1P, β-catenin, and axin-2 expression, followed with improvement in tissue damage. © 2024 Salbiah Ridwan et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/). All Rights Reserved.