Exploring the potential molecular targets of hydroxymethylbutyrate and glucosamine fortified whey protein drink to modulate sarcopenia and Alzheimer's disease by in silico and in vitro studies

Fortified foods have garnered significant attention as potential therapeutic strategies with less adverse effects and ready accessibility. However, precise formulation is required to optimize their beneficial effects. Our study aimed to unravel the mechanisms of a functional protein beverage, hydroxymethylbutyrate and glucosamine fortified whey protein drink (HG-WPD), as a possible intervention combatting sarcopenia and Alzheimer's disease (AD) by network pharmacology, molecular docking, and experimental assays. This study investigates the molecular pathways through which HG-WPD acts, by combining in silico and in vitro analyses. Our in silico study predicted the important target proteins, including acetylcholinesterase (AChE), matrix metalloproteinase 9 (MMP9), and angiotensin-converting enzyme (ACE), linked to both diseases. The molecular docking analysis showed that hydroxymethylbutyrate and glucosamine exhibited a notable binding affinity to these proteins, therefore suggesting their possible use as multi-target medicinal agents. In vitro studies on C2C12 myotubes showed that HG-WPD reduced dexamethasone-induced cell death highlighting its potential anti-sarcopenic actions. Furthermore, confirming their possible function in reducing cognitive impairment in AD, the antioxidant tests showed great activity of both substances. The results imply that HG-WPD is potentially effective for future therapeutic approaches as a functional food product with dual benefits in combatting sarcopenia and AD.