TY - JOUR
T1 - Identification of Small-Molecule Inhibitors of Human Inositol Hexakisphosphate Kinases by High-Throughput Screening
AU - Liao, Gangling
AU - Ye, Wenjuan
AU - Heitmann, Tyler
AU - Ernst, Glen
AU - Depasquale, Michael
AU - Xu, Laiyi
AU - Wormald, Michael
AU - Hu, Xin
AU - Ferrer, Marc
AU - Harmel, Robert K.
AU - Fiedler, Dorothea
AU - Barrow, James
AU - Wei, Huijun
N1 - Funding Information:
This work was fund by the Lieber Institute for Brain Development and U01 MH112658 "01A1 from NIH/NIMH and the NIH Intramural Research Program to NCATS.
Publisher Copyright:
© 2021 American Chemical Society.
PY - 2021/4/9
Y1 - 2021/4/9
N2 - Inositol hexakisphosphate kinases (IP6Ks) catalyze pyrophosphorylation of inositol hexakisphosphate (IP6) into inositol 5-diphospho-1,2,3,4,6-pentakisphosphate (IP7), which is involved in numerous areas of cell physiology including glucose homeostasis, blood coagulation, and neurological development. Inhibition of IP6Ks may be effective for the treatment of Type II diabetes, obesity, metabolic complications, thrombosis, and psychiatric disorders. We performed a high-throughput screen (HTS) of 158 »410 compounds for IP6K1 inhibitors using a previously developed ADP-Glo Max assay. Of these, 1206 compounds were found to inhibit IP6K1 kinase activity by more than 25%, representing a 0.8% hit rate. Structural clustering analysis of HTS-active compounds, which were confirmed in the dose-response testing using the same kinase assay, revealed diverse clusters that were feasible for future structure-activity relationship (SAR) optimization to potent IP6K inhibitors. Medicinal chemistry SAR efforts in three chemical series identified potent IP6K1 inhibitors which were further validated in an orthogonal LC-MS IP7 analysis. The effects of IP6K1 inhibitors on cellular IP7 levels were further confirmed and were found to correlate with cellular IP6K1 binding measured by a high-throughput cellular thermal shift assay (CETSA).
AB - Inositol hexakisphosphate kinases (IP6Ks) catalyze pyrophosphorylation of inositol hexakisphosphate (IP6) into inositol 5-diphospho-1,2,3,4,6-pentakisphosphate (IP7), which is involved in numerous areas of cell physiology including glucose homeostasis, blood coagulation, and neurological development. Inhibition of IP6Ks may be effective for the treatment of Type II diabetes, obesity, metabolic complications, thrombosis, and psychiatric disorders. We performed a high-throughput screen (HTS) of 158 »410 compounds for IP6K1 inhibitors using a previously developed ADP-Glo Max assay. Of these, 1206 compounds were found to inhibit IP6K1 kinase activity by more than 25%, representing a 0.8% hit rate. Structural clustering analysis of HTS-active compounds, which were confirmed in the dose-response testing using the same kinase assay, revealed diverse clusters that were feasible for future structure-activity relationship (SAR) optimization to potent IP6K inhibitors. Medicinal chemistry SAR efforts in three chemical series identified potent IP6K1 inhibitors which were further validated in an orthogonal LC-MS IP7 analysis. The effects of IP6K1 inhibitors on cellular IP7 levels were further confirmed and were found to correlate with cellular IP6K1 binding measured by a high-throughput cellular thermal shift assay (CETSA).
KW - IP7
KW - cellular thermal shift assay (CETSA)
KW - high-throughput screen
KW - inositol hexakisphosphate kinases
KW - inositol pyrophosphate
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U2 - 10.1021/acsptsci.0c00218
DO - 10.1021/acsptsci.0c00218
M3 - Article
C2 - 33860201
AN - SCOPUS:85103442802
SN - 2575-9108
VL - 4
SP - 780
EP - 789
JO - ACS Pharmacology and Translational Science
JF - ACS Pharmacology and Translational Science
IS - 2
ER -