Targeting PI3K Signaling for Broad Inhibition of β-Coronavirus Infection

The phosphatidylinositol 3-kinase (PI3K) signaling pathway plays a central role in regulating key cellular processes such as survival, metabolism, and immune responses. Aberrant activation of this pathway is associated with tumorigenesis, and several PI3K inhibitors have been developed as anticancer agents. Emerging evidence suggests that viruses, including β-coronaviruses, have evolved mechanisms to exploit host PI3K signaling for their replication and immune evasion. In this study, we evaluated the antiviral efficacy of a panel of PI3K inhibitors against β-coronaviruses, including mouse hepatitis virus (MHV), human OC43 (HuCoV-OC43) and four major SARS-CoV-2 variants using both cell line and organoid models. Our findings reveal that these compounds exhibit low micromolar potency in inhibiting viral replication. Notably, the inhibitor C20 (PWT33597) demonstrated broad-spectrum activity against multiple β-coronaviruses, including SARS-CoV-2, MHV, and HuCoV-OC43, in conventional cell lines as well as in air–liquid interface (ALI)-cultured, differentiated primary human nasal and bronchial epithelial cells. Given that cytokine storm is a major contributor to SARS-CoV-2–related multiorgan failure and mortality, we further explored the impact of PI3K inhibition on host inflammatory responses. We found that MHV infection markedly increased cytokine expression in 17CL-1 fibroblasts and RAW264.7 macrophages. Interestingly, treatment with C20 further amplified cytokine production in this context, suggesting complex immunomodulatory effects that warrant further investigation. Together, our findings support the therapeutic potential of repurposing PI3K inhibitors as broad-spectrum antivirals. These compounds not only suppress viral replication but may also influence host immune responses, providing a promising avenue for intervention against current and emerging coronavirus threats.