Identification of Small-Molecule Inhibitors of Human Inositol Hexakisphosphate Kinases by High-Throughput Screening

Area of Science:

• Biochemistry
• Enzymology
• Medicinal Chemistry

Background:

• Inositol hexakisphosphate kinases (IP6Ks) produce inositol 5-diphospho-1,2,3,4,6-pentakisphosphate (IP7), crucial for glucose homeostasis, blood coagulation, and neurological functions.
• IP6K inhibition offers therapeutic potential for Type II diabetes, obesity, metabolic disorders, thrombosis, and psychiatric conditions.

Purpose of the Study:

• To identify novel inhibitors of Inositol hexakisphosphate kinase 1 (IP6K1) through a high-throughput screening (HTS) campaign.
• To explore the potential of IP6K1 inhibitors for therapeutic applications in metabolic and neurological diseases.

Main Methods:

• A high-throughput screen (HTS) of 158,410 compounds using an ADP-Glo Max assay to identify IP6K1 inhibitors.
• Dose-response testing, structural clustering, medicinal chemistry SAR optimization, LC-MS IP7 analysis, and cellular thermal shift assay (CETSA) for validation.

Main Results:

• A hit rate of 0.8% was observed, with 1206 compounds inhibiting IP6K1 activity by over 25%.
• Structural analysis revealed diverse compound clusters suitable for optimization.
• Medicinal chemistry efforts yielded potent IP6K1 inhibitors, validated by reduced cellular IP7 levels and correlated with IP6K1 binding via CETSA.

Conclusions:

• The HTS campaign successfully identified diverse chemical scaffolds with IP6K1 inhibitory activity.
• The identified inhibitors demonstrate potential for further development as therapeutics for IP6K-related diseases.
• Cellular assays confirmed the efficacy of IP6K1 inhibitors in modulating IP7 levels and binding to the target protein.