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Salicylate esters inhibit human and rat 11β-Hydroxysteroid dehydrogenase type 2: Mechanistic insights from enzyme
Yinwei Dai1, Zhuoqi Chen2, Xiya Ren2
1Department of Anesthesiology and Perioperative Medicine, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China; Department of Obstetrics and Gynecology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China; Key Laboratory of Pediatric Anesthesiology, Ministry of Education, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China; Key Laboratory of Precision Anesthesiology of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China.
Abstract:
Salicylate esters are widely used in foods, pharmaceuticals, and cosmetics, but their effects on 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2), a key enzyme in cortisol metabolism and placental glucocorticoid protection, remain unclear. We investigated the inhibitory effects of 15 salicylate esters on human and rat 11β-HSD2 using enzyme kinetics, SPR, cellular assays, SAR/QSAR, docking, molecular dynamics, and network toxicology. Benzyl salicylate was the most potent inhibitor, with IC50 values of 18.53 μM for human and 29.25 μM for rat 11β-HSD2. Global nonlinear fitting with AICc-based model comparison supported predominantly noncompetitive or mixed/noncompetitive inhibition of human 11β-HSD2 with respect to cortisol, whereas rat 11β-HSD2 showed compound-dependent apparent inhibition patterns; Lineweaver-Burk plots were retained only for visualization. SPR showed direct binding of benzyl and homomenthyl salicylates to human 11β-HSD2, with KD values of 21.6 and 47.4 μM, respectively, and suggested that benzyl salicylate interferes with NAD+ binding. Docking localized active salicylates near the NAD+/steroid interface, and molecular dynamics supported complex stability. In BeWo cells, active salicylates reduced cortisone production under largely noncytotoxic conditions, supporting impaired cortisol metabolism in intact cells. SAR and 3D-QSAR analyses identified lipophilicity, steric features, and aromaticity as major determinants of potency, while exploratory network toxicology suggested possible links to placental function and fetal growth-related pathways. These findings identify salicylate esters as 11β-HSD2 inhibitors and provide mechanistic evidence for their potential interference with glucocorticoid metabolism in vitro.
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