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Published on: August 19, 2021
Establishment of hypoglycemic agent screening method based on human glucokinase
Chou-Fei Wu1, Yang Xu, Yong Tao
1Key Laboratory of State Food Science and Technology, Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, Jiangxi, China.
Biomedical and Environmental Sciences : BES
|May 26, 2009
Summary
A new in vitro platform reliably screens glucokinase activators (GKAs). Optimized conditions using response surface methodology (RSM) achieved a 34.8% activity ratio, validating the model for GKA discovery.
Area of Science:
- Biochemistry
- Enzyme kinetics
- Drug discovery
Background:
- Glucokinase (GCK) plays a crucial role in glucose homeostasis and is a therapeutic target for diabetes.
- Developing efficient screening platforms is essential for identifying novel glucokinase activators (GKAs).
Purpose of the Study:
- To establish and optimize a reliable in vitro platform for screening glucokinase activators (GKAs).
Main Methods:
- Production of histidine-tagged human pancreatic glucokinase (PGK) using a prokaryotic expression system.
- Optimization of a microplate-based GKA screening platform using response surface methodology (RSM).
- Plackett-Burman design (PBD) identified initial pH, reaction time, and MgCl2 as key factors; a central composite design (CCD) further refined these parameters.
Main Results:
- Optimal conditions determined were pH 7.0, reaction time of 13.7 min, and MgCl2 concentration of 2.11 mmol/L.
- The predicted maximum activity ratio was 34.1%, with an experimentally achieved ratio of 34.8%.
- The model demonstrated high credibility with a determination coefficient (R2) of 0.9442.
Conclusions:
- The optimized platform provides a reliable method for screening GKAs.
- LLAE3, a compound from Folium nelumbinis, demonstrated significant PGK activation under optimal conditions.
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