*Article* **Therapeutic Potential of Deflamin against Colorectal Cancer Development and Progression**

**Sara Silva 1,†, Ana Cavaco 1,†, Bianca Basso 1, Joana Mota 2,3, Raquel Cruz-Duarte 1, Miguel Costa 1, Lara Carvalho 1, Ana Lima 2,3,\*, Luis Costa 1,4, Ricardo Ferreira 3,‡ and Marta Martins 1,\*,‡**


**Simple Summary:** We have previously identified deflamin, an oligomeric protein isolated from the white lupine seeds (*Lupinus albus*) with anti-MMPs and anti-inflammatory properties. Given the involvement of MMPs and inflammation in the carcinogenesis process, we aimed to assess deflamin's role in cancer development and progression. Using colorectal cancer cell lines and zebrafish xenotransplant models, we found that deflamin exhibits anti-MMP-2 and anti-MMP-9 activity, being able to reduce tumor size and metastasis formation in vivo. Deflamin was shown to impair cancer cell migration and invasion, as well as collagen remodeling and angiogenesis in the tumor microenvironment, highly impacting cancer behavior. Overall, our results unravel the nutraceutical potential of deflamin in colorectal cancer treatment.

**Abstract:** Matrix metalloproteinases (MMPs) are proteolytic enzymes that play a crucial role in tumor microenvironment remodeling, contributing to inflammatory and angiogenic processes, and ultimately promoting tumor maintenance and progression. Several studies on bioactive polypeptides isolated from legumes have shown anti-migratory, anti-MMPs, and anti-tumor effects, potentially constituting novel strategies for both the prevention and progression of cancer. In this work, we investigated the anti-tumor role of deflamin, a protein oligomer isolated from white lupine seeds (*Lupinus albus*) reported to inhibit MMP-9 and cell migration in colorectal cancer (CRC) cell lines. We found that deflamin exerts an inhibitory effect on tumor growth and metastasis formation, contributing to increased tumor apoptosis in the xenotransplanted zebrafish larvae model. Furthermore, deflamin resulted not only in a significant reduction in MMP-2 and MMP-9 activity but also in impaired cancer cell migration and invasion in vitro. Using the xenograft zebrafish model, we observed that deflamin inhibits collagen degradation and angiogenesis in the tumor microenvironment in vivo. Overall, our work reveals the potential of deflamin as an agent against CRC development and progression.

**Keywords:** deflamin; colorectal cancer; MMP-2; MMP-9
