Metastatic colorectal carcinoma (mCRC) is one of the prevalent subtypes of human cancers and is caused by the alterations of various lifestyle and diet‐associated factors. β‐catenin, GSK‐3β, PI3K‐α, AKT1, and NF‐κB p50 are known to be the critical regulators of tumorigenesis and immunopathogenesis of mCRC. Unfortunately, current drugs have limited efficacy, side effects and can lead to chemoresistance. Therefore, searching for a nontoxic, efficacious anti‐mCRC agent is crucial and of utmost interest. The present study demonstrates the identification of a productive and nontoxic anti‐mCRC agent through a five‐targets (β‐catenin, GSK‐3β, PI3K‐α, AKT1, and p50)‐based and three‐tier (binding affinity, pharmacokinetics, and pharmacophore) screening strategy involving a series of 30 phytocompounds having a background of anti‐inflammatory/anti‐mCRC efficacy alongside 5‐fluorouracil (FU), a reference drug. Luteolin (a phyto‐flavonoid) was eventually rendered as the most potent and safe phytocompound. This inference was verified through three rounds of validation. Firstly, luteolin was found to be effective against the different mCRC cell lines (HCT‐15, HCT‐116, DLD‐1, and HT‐29) without hampering the viability of non‐tumorigenic ones (RWPE‐1). Secondly, luteolin was found to curtail the clonogenicity of CRC cells, and finally, it also disrupted the formation of colospheroids, a characteristic of metastasis. While studying the mechanistic insights, luteolin was found to inhibit β‐catenin activity (a key regulator of mCRC) through direct physical interactions, promoting its degradation by activating GSK3‐β and ceasing its activation by inactivating AKT1 and PI3K‐α. Luteolin also inhibited p50 activity, which could be useful in mitigating mCRC‐associated proinflammatory milieu. In conclusion, our study provides evidence on the efficacy of luteolin against the critical key regulators of immunopathogenesis of mCRC and recommends further studies in animal models to determine the effectiveness efficacy of this natural compound for treating mCRC in the future.
