Journal
SLAS DISCOVERY
Volume 25, Issue 9, Pages 1038-1046Publisher
ELSEVIER SCIENCE INC
DOI: 10.1177/2472555220926146
Keywords
aldolase A; luminescence; NADH; anticancer; high-throughput screening
Funding
- CPRIT [RP110532-P1, RP160657]
- Florida Drug Discovery Acceleration Program (State of Florida, Department of Health)
- NIH [R01 CA216424]
Ask authors/readers for more resources
Hypoxic solid tumors induce the stabilization of hypoxia-inducible factor 1 alpha (HIF1 alpha), which stimulates the expression of many glycolytic enzymes and hypoxia-responsive genes. A high rate of glycolysis supports the energetic and material needs for tumors to grow. Fructose-1,6-bisphosphate aldolase A (ALDOA) is an enzyme in the glycolytic pathway that promotes the expression of HIF1 alpha. Therefore, inhibition of ALDOA activity represents a potential therapeutic approach for a range of cancers by blocking two critical cancer survival mechanisms. Here, we present a luminescence-based strategy to determine ALDOA activity. The assay platform was developed by integrating a previously established ALDOA activity assay with a commercial NAD/NADH detection kit, resulting in a significant (>12-fold) improvement in signal/background (S/B) compared with previous assay platforms. A screening campaign using a mixture-based compound library exhibited excellent statistical parameters of Z ' (>0.8) and S/B (similar to 20), confirming its robustness and readiness for high-throughput screening (HTS) application. This assay platform provides a cost-effective method for identifying ALDOA inhibitors using a large-scale HTS campaign.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available