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Screening of 5- and 6-substituted amiloride libraries identifies dual-UPA/NHE1 active and single target-selective inhibitors

Journal Article


Abstract


  • The K+-sparing diuretic amiloride shows off-target anti-cancer effects in multiple rodent models. These effects arise from the inhibition of two distinct cancer targets: the trypsin-like serine protease urokinase-type plasminogen activator (uPA), a cell-surface mediator of matrix degradation and tumor cell invasiveness, and the sodium-hydrogen exchanger isoform-1 (NHE1), a central reg-ulator of transmembrane pH that supports carcinogenic progression. In this study, we co-screened our library of 5- and 6-substituted amilorides against these two targets, aiming to identify single-target selective and dual-targeting inhibitors for use as complementary pharmacological probes. Closely related analogs substituted at the 6-position with pyrimidines were identified as dual-tar-geting (pyrimidine 24 uPA IC50 = 175 nM, NHE1 IC50 = 266 nM, uPA selectivity ratio = 1.5) and uPA-selective (methoxypyrimidine 26 uPA IC50 = 86 nM, NHE1 IC50 = 12,290 nM, uPA selectivity ratio = 143) inhibitors, while high NHE1 potency and selectivity was seen with 5-morpholino (29 NHE1 IC50 = 129 nM, uPA IC50 = 10,949 nM; NHE1 selectivity ratio = 85) and 5-(1,4-oxazepine) (30 NHE1 IC50 = 85 nM, uPA IC50 = 5,715 nM; NHE1 selectivity ratio = 67) analogs. Together, these amilorides comprise a new toolkit of chemotype-matched, non-cytotoxic probes for dissecting the pharmacological effects of selective uPA and NHE1 inhibition versus dual-uPA/NHE1 inhibition.

Publication Date


  • 2021

Citation


  • Buckley, B. J., Kumar, A., Aboelela, A., Bujaroski, R. S., Li, X., Majed, H., . . . Kelso, M. J. (2021). Screening of 5- and 6-substituted amiloride libraries identifies dual-UPA/NHE1 active and single target-selective inhibitors. International Journal of Molecular Sciences, 22(6), 1-12. doi:10.3390/ijms22062999

Scopus Eid


  • 2-s2.0-85102440481

Start Page


  • 1

End Page


  • 12

Volume


  • 22

Issue


  • 6

Abstract


  • The K+-sparing diuretic amiloride shows off-target anti-cancer effects in multiple rodent models. These effects arise from the inhibition of two distinct cancer targets: the trypsin-like serine protease urokinase-type plasminogen activator (uPA), a cell-surface mediator of matrix degradation and tumor cell invasiveness, and the sodium-hydrogen exchanger isoform-1 (NHE1), a central reg-ulator of transmembrane pH that supports carcinogenic progression. In this study, we co-screened our library of 5- and 6-substituted amilorides against these two targets, aiming to identify single-target selective and dual-targeting inhibitors for use as complementary pharmacological probes. Closely related analogs substituted at the 6-position with pyrimidines were identified as dual-tar-geting (pyrimidine 24 uPA IC50 = 175 nM, NHE1 IC50 = 266 nM, uPA selectivity ratio = 1.5) and uPA-selective (methoxypyrimidine 26 uPA IC50 = 86 nM, NHE1 IC50 = 12,290 nM, uPA selectivity ratio = 143) inhibitors, while high NHE1 potency and selectivity was seen with 5-morpholino (29 NHE1 IC50 = 129 nM, uPA IC50 = 10,949 nM; NHE1 selectivity ratio = 85) and 5-(1,4-oxazepine) (30 NHE1 IC50 = 85 nM, uPA IC50 = 5,715 nM; NHE1 selectivity ratio = 67) analogs. Together, these amilorides comprise a new toolkit of chemotype-matched, non-cytotoxic probes for dissecting the pharmacological effects of selective uPA and NHE1 inhibition versus dual-uPA/NHE1 inhibition.

Publication Date


  • 2021

Citation


  • Buckley, B. J., Kumar, A., Aboelela, A., Bujaroski, R. S., Li, X., Majed, H., . . . Kelso, M. J. (2021). Screening of 5- and 6-substituted amiloride libraries identifies dual-UPA/NHE1 active and single target-selective inhibitors. International Journal of Molecular Sciences, 22(6), 1-12. doi:10.3390/ijms22062999

Scopus Eid


  • 2-s2.0-85102440481

Start Page


  • 1

End Page


  • 12

Volume


  • 22

Issue


  • 6