Cocrystal-inspired glycyrrhizic acid–azole nanoassemblies for synergistic biofilm disruption and immune modulation in fungal infections

Fungal biofilms represent a major therapeutic hurdle due to their drug resistance and immune evasion. Here, we report a cocrystal-inspired nano-assembly strategy that co-assembles glycyrrhizic acid (GA), a natural triterpenoid saponin, with azole antifungal agents into multifunctional nanoparticles. The GA-azole co-assemblies, stabilized via hydrogen bonding and hydrophobic interactions, exhibit cocrystal-like properties while preserving the bioactivities of both components. This hybrid nanoplatform improves azole solubility and permeability and leverages GA's intrinsic membrane-disrupting, ROS-scavenging, and immunomodulatory effects. Mechanistic studies revealed that the nanoparticles effectively disrupted Candida albicans biofilms by impairing matrix structure and suppressing hyphal formation. Transcriptomic and qRT-PCR analyses demonstrated GA-azole co-assemblies downregulated genes involved in ergosterol biosynthesis, oxidative stress response, and morphogenesis. Additionally, GA-azole nanoparticles promoted macrophage polarization toward an anti-inflammatory M2 phenotype and alleviated oxidative stress, by activating the Nrf2/HO-1 antioxidant pathway, thereby modulating the excessive inflammation stimulated by fungal infection. In vitro and in vivo experiments, including a murine perianal infection model, showed significant reductions in fungal burden, biofilm thickness, and local inflammation without systemic toxicity. This work presents a multi-targeted nanotherapeutic strategy combining enhanced drug delivery, antifungal synergy, immune regulation to combat biofilm-associated fungal infections.