Recognized for its dual nutritional and therapeutic value, the fungus Hericium erinaceus is increasingly acknowledged as a rich resource of naturally derived α-glucosidase inhibitors. In this study, eight compounds were isolated from H. erinaceus, including three novel compounds designated as erinacerins X−Z (1, 2, and 6). Their absolute configurations were definitively elucidated using a combination of NMR, HR-MS, and ECD calculations. Furthermore, an integrated screening strategy combining molecular docking, molecular dynamics (MD) simulations, and surface plasmon resonance (SPR) analysis identified two isoindolin-1-ones (2 and 3) as potent naturally derived α-glucosidase inhibitors. Notably, in vitro testing established compounds 2 and 3 as robust α-glucosidase inhibitors, affording IC₅₀ values of 17.80 ± 1.03 μM and 19.50 ± 1.33 μM, respectively. MD simulations revealed that electrostatic interactions and van der Waals forces are the primary drivers of this intermolecular association. These findings were further corroborated by SPR analysis, which quantified their high-affinity binding kinetics to the enzyme. Overall, this combined approach establishes a solid foundation for the discovery and development of natural α-glucosidase inhibitors from H. erinaceus.

To address the lack of a comprehensive quality control system for Ganoderma lucidum, we developed an integrated evaluation strategy across four varieties and three growth stages. This system integrates the targeted screening of anti-benign prostatic hyperplasia (BPH) triterpenoids acting on 5α-reductase type 2 (SRD5A2) with a chemical consistency assessment utilizing systematic quantitative fingerprint method (SQFM) and chemometrics. UPLC-Q-TOF-MS/MS identified 85 triterpenoids in the samples. An orthogonal partial least squares (OPLS) regression was utilized to screen seven chromatographic peaks that positively correlated with SRD5A2 inhibitory activity. Three principal bioactives were structurally identified as ganoderic acids DM and B, and ganoderenic acid A, which demonstrated significant in vitro SRD5A2 inhibition rates of 61.16 ± 1.87%, 36.41 ± 1.10%, and 41.82 ± 2.09%, respectively. Molecular dynamics simulations and averaged weak interaction analysis revealed that these compounds exert potent enzyme inhibition via hydrogen bonds and hydrophobic interactions with distinct SRD5A2 amino acid residues. The SQFM-chemometrics quality system confirmed ten samples reached Grade 6 or above, identifying the H3 variety at the initial stage as possessing the highest active ingredient content and optimal overall quality. This integrated framework enables rapid bioactive discovery and robust standardization for G. lucidum-based functional foods, thereby facilitating their industrial development.