11β-Hydroxysteroid Dehydrogenase Type 1 Inhibitor Development by Lentiviral Screening Based on Computational...

Area of Science:

• Biochemistry
• Molecular Biology
• Pharmacology

Background:

• 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) is a key enzyme in regulating glucocorticoid activity.
• Dysregulation of 11β-HSD1 is implicated in metabolic disorders such as obesity and type 2 diabetes.

Purpose of the Study:

• To establish stable cell lines expressing rat and human 11β-HSD1 for drug discovery.
• To identify potential inhibitors of 11β-HSD1 using computational and experimental methods.

Main Methods:

• Lentiviral transduction was used to express rat and human 11β-HSD1 in CHO-K1 cells.
• Catalytic activity was measured using a scanning radiometer.
• Molecular docking simulations (AutoDock, LeDock) predicted inhibitor-enzyme interactions.
• In vivo studies involved C57BL/6 mice with R11bHsd1 overexpression.

Main Results:

• Stable expression of functional rat and human 11β-HSD1 in CHO-K1 cells was achieved with high catalytic activity (99.5% and 98.7%).
• Compound 8 demonstrated potential as an 11β-HSD1 inhibitor by forming hydrogen bonds with key enzyme residues.
• Overexpression of R11bHsd1 in mice led to increased body weight and metabolic parameters.

Conclusions:

• Stable 11β-HSD1-expressing cell lines are suitable for high-throughput screening of inhibitors.
• Compound 8 is a promising candidate for further development as an 11β-HSD1 inhibitor.
• These findings provide a foundation for developing novel therapeutic strategies targeting 11β-HSD1.