Exploration of Baicalein-Core Derivatives as Potent Antifungal Agents: SAR and Mechanism Insights

HIGHLIGHTS: What are the main findings? Baicalein-Core Derivatives were designed and synthesized as Potent Anti-Fluconazole-resistant fungal Agents. O-dihydroxyls and vic-trihydroxy groups on either the A ring or B ring of flavones play a crucial role. MoA: Inhibit hypha formation; Little effect on ergosterol biosynthesis; Weak inhibitory effect on Eno1. Potential targets: 1,3-β-d-glucan synthase catalytic subunit, 1,3-β-d-glucan-UDP glucosyltransferase, and glycosyl-phosphatidylinositol protein by in-silico assay. What is the implication of the main finding? Provide potential synergistic antifungals with new MoA. ABSTRACT: Baicalein (BE), the major component of Scutellaria Baicalensis, exhibited potently antifungal activity against drug-resistant Candida albicans, and strong inhibition on biofilm formation. Therefore, a series of baicalein-core derivatives were designed and synthesized to find more potent compounds and investigate structure–activity relationship (SAR) and mode of action (MoA). Results demonstrate that A4 and B5 exert a more potent antifungal effect (MIC(80) = 0.125 μg/mL) than BE (MIC(80) = 4 μg/mL) when used in combination with fluconazole (FLC), while the MIC(80) of FLC dropped from 128 μg/mL to 1 μg/mL. SAR analysis indicates that the presence of 5-OH is crucial for synergistic antifungal activities, while o-dihydroxyls and vic-trihydroxyls are an essential pharmacophore, whether they are located on the A ring or the B ring of flavonoids. The MoA demonstrated that these compounds exhibited potent antifungal effects by inhibiting hypha formation of C. albicans. However, sterol composition assay and enzymatic assay conducted in vitro indicated minimal impact of these compounds on sterol biosynthesis and Eno1. These findings were further confirmed by the results of the in-silico assay, which assessed the stability of the complexes. Moreover, the inhibition of hypha of this kind of compound could be attributed to their effect on the catalytic subunit of 1,3-β-d-glucan synthase, 1,3-β-d-glucan-UDP glucosyltransferase and glycosyl-phosphatidylinositol protein, rather than inhibiting ergosterol biosynthesis and Eno1 activity by Induced-Fit Docking and Molecular Dynamics Simulations. This study presents potential antifungal agents with synergistic effects that can effectively inhibit hypha formation. It also provides new insights into the MoA.