Unraveling molecular mechanisms of β-glucuronidase inhibition by flavonoids from Centaurea scoparia: integrated in silico and in vitro insights
Investigating the detailed molecular mechanisms of β-glucuronidase inhibition is critical for pioneering new therapeutic solutions and driving progress in pharmaceutical research. The inhibitory potential of four flavonoid aglycones derived from Centaurea scoparia against β-glucuronidase was thoroug...
| Autores: | , , , , , , |
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| Tipo de recurso: | artículo |
| Fecha de publicación: | 2024 |
| País: | España |
| Institución: | Universidad Autónoma de Madrid |
| Repositorio: | Biblos-e Archivo. Repositorio Institucional de la UAM |
| Idioma: | inglés |
| OAI Identifier: | oai:repositorio.uam.es:10486/720837 |
| Acceso en línea: | http://hdl.handle.net/10486/720837 https://dx.doi.org/10.1039/d4nj02393e |
| Access Level: | acceso abierto |
| Palabra clave: | β-Glucuronidase In Vitro study Enzyme inhibition Molecular dynamic simulations Enzyme kinetics Química |
| Sumario: | Investigating the detailed molecular mechanisms of β-glucuronidase inhibition is critical for pioneering new therapeutic solutions and driving progress in pharmaceutical research. The inhibitory potential of four flavonoid aglycones derived from Centaurea scoparia against β-glucuronidase was thoroughly examined using a combination of in vitro and in silico methodologies. The in vitro assays revealed that compounds 1 and 3 had the strongest inhibitory effects, demonstrated by their low IC<inf>50</inf> values of 3.16 ± 0.34 and 3.82 ± 0.1 μM, respectively. The results of the enzyme kinetics assay revealed that compounds 2 and 3, and the reference drug EGCG displayed a mixed inhibition mode. Conversely, compound 1 was found to operate via a noncompetitive inhibition mechanism, as evidenced by the intersection patterns in the Lineweaver-Burk plots. The outcomes from the docking analysis are consistent with the in vitro inhibitory efficacy assays, showing that compounds 1 and 3 possess the lowest binding affinities (−8.6 and −9.0 kcal mol<sup>−1</sup>, respectively). Isolated phytochemicals demonstrated substantial polar and hydrophobic interactions with the residues within the enzyme's binding site. We investigated the interaction dynamics of isolated compounds with β-glucuronidase using a 100 ns molecular dynamics (MD) simulation. The analysis of various MD parameters indicated that compounds 1 and 3 showed stable trajectories and significant energy stabilization when bound to β-glucuronidase. Furthermore, compounds 1 and 3 exhibited the most favorable average Coulombic short-range interaction energies, recorded at −86.53 ± 11 kJ mol<sup>−1</sup> and −98.04 ± 17 kJ mol<sup>−1</sup>, respectively. These compounds also demonstrated the lowest average Lennard-Jones short-range interaction energies, measured at −84.56 ± 14 kJ mol<sup>−1</sup> and −106.02 ± 4.6 kJ mol<sup>−1</sup>, respectively. The results of MM/PBSA calculations revealed binding free energies of −10.99 ± 2.07, −11.39 ± 2.71, −26.77 ± 3.83, and 0.20 ± 0.32 kJ mol<sup>−1</sup> for isolated compounds 1-4 with the target enzyme, respectively. These computational results support the experimental data, suggesting that compounds 1 and 3 could be potent inhibitors of β-glucuronidase |
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