EphA2 blockage ALW-II-41-27 alleviates atherosclerosis by remodeling gut microbiota to regulate bile acid metabolism

Coronary artery disease (CAD) remains a critical global health challenge driven by systemic inflammation, metabolic dysfunction, and gut microbiota dysbiosis. ALW-II-41-27, a specific inhibitor of the EphA2 receptor, has demonstrated anti-inflammatory properties; however, its role in atherosclerosis has not been fully explored. This study investigated the therapeutic potential of pharmacologically inhibiting EphA2 with ALW-II-41-27 and the underlying mechanisms in mitigating atherosclerosis.

ALW-II-41-27 was administered via intraperitoneal injection to apoE-/- mice on a high-fat diet. Treatment significantly reduced atherosclerotic plaque burden, characterized by decreased lipid and macrophage accumulation and increased collagen and smooth muscle cell content. Plasma and hepatic cholesterol levels were also notably reduced, along with attenuation of colonic inflammation.

Gut microbiota analysis via metagenomics revealed that ALW-II-41-27 enriched beneficial bacterial genera such as Enterococcus, Akkermansia, Eggerthella, and Lactobacillus, which were associated with enhanced production of secondary bile acids. Untargeted metabolomics of plasma metabolites corroborated these findings. To establish a causal relationship between ALW II-41-27 gut microbiota changes and atherosclerosis, fecal microbiota transplantation was conducted. Mice receiving feces from ALW-II-41-27-treated donors exhibited reduced atherosclerotic plaque burden.

In a clinical cohort, CAD patients exhibited lower plasma levels of deoxycholic acid (DCA) and hyodeoxycholic acid (HDCA), which correlated negatively with higher monocyte EphA2 expression. These findings suggest that ALW-II-41-27 exerts its anti-atherosclerotic effects by modulating gut microbiota composition and bile acid metabolism. The results highlight ALW-II-41-27 as a promising therapeutic agent for CAD, offering novel insights into the interplay between EphA2 inhibition, gut microbiota, and systemic metabolic pathways.