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In this study, the synergistic inhibition of \u03b1-GLU by hyperin, isoquercitrin, and quercetin was systematically investigated using fluorescence quenching, isothermal titration calorimetry, circular dichroism, differential scanning calorimetry, computational models, and Caco-2 cell assays. The ternary combination synergistically inhibited \u03b1-GLU (IC50 = 0.022 mg\/mL, combination index &amp;lt;1) and showed the greatest \u03b1-helix reduction and denaturation temperature decrease of \u03b1-GLU. Molecular docking and molecular dynamics simulations suggested that quercetin preferentially occupied the hydrophobic catalytic pocket, whereas hyperin and isoquercitrin primarily interacted with peripheral regions. In differentiated Caco-2 cells, hyperin and isoquercitrin exhibited stronger inhibition toward sucrase than maltase, whereas quercetin showed the opposite trend. Collectively, the results demonstrate that the synergistic inhibition of \u03b1-GLU arises from complementary binding modes, cooperative conformational perturbation, and differential targeting of intestinal disaccharidase activities.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c07937","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T10:01:49Z","timestamp":1789120909000},"page":"29297-29312","source":"Crossref","is-referenced-by-count":0,"title":["Synergistic Inhibition\nof \u03b1-Glucosidase\nby Hyperin, Isoquercitrin, and Quercetin: Mechanistic Insights from\nMulti-spectroscopic, Calorimetric, Computational, and Caco-2\nCell Assays"],"prefix":"10.1021","volume":"74","author":[{"given":"Di","family":"Zhao","sequence":"first","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruona","family":"Li","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiayi","family":"Hu","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang","family":"Wang","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]},{"name":"Hainan Institute of Northwest A&F University , , ,","place":["Sanya, Hainan, China, 572025"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingwei","family":"Hou","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]},{"name":"Hainan Institute of Northwest A&F University , , ,","place":["Sanya, Hainan, China, 572025"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6769-4663","authenticated-orcid":true,"given":"Mei","family":"Li","sequence":"additional","affiliation":[{"name":"Northwest A&F University , , ,","place":["Yangling, China, 712100"]},{"name":"Hainan Institute of Northwest A&F University , , ,","place":["Sanya, Hainan, China, 572025"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,11]]},"reference":[{"issue":"26","key":"2026092310084275100_cit1","doi-asserted-by":"publisher","first-page":"2488","DOI":"10.2174\/0115680266260682230921054652","article-title":"An Updated Overview of Synthetic\n\u03b1-Glucosidase Inhibitors: Chemistry and Bioactivities","volume":"23","author":"Cai","year":"2023","journal-title":"Curr. 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