In Silico Assisted Identification, Synthesis, and In Vitro Pharmacological Characterization of Potent and Selective Blockers of the Epilepsy-Associated KCNT1 Channel

  • J Med Chem. 2024 Jun 13;67(11):9124-9149. doi: 10.1021/acs.jmedchem.4c00268.
Nunzio Iraci  1 ,  Lidia Carotenuto  2 ,  Tania Ciaglia  3 ,  Giorgio Belperio  4 ,  Francesca Di Matteo  3 ,  Ilaria Mosca  5 ,  Giusy Carleo  2 ,  Manuela Giovanna Basilicata  6 ,  Paolo Ambrosino  4 ,  Rita Turcio  3 ,  Deborah Puzo  5 ,  Giacomo Pepe  3 ,  Isabel Gomez-Monterrey  7 ,  Maria Virginia Soldovieri  5 ,  Veronica Di Sarno  3 ,  Pietro Campiglia  3 ,  Francesco Miceli  2 ,  Alessia Bertamino  3 ,  Carmine Ostacolo  3 ,  Maurizio Taglialatela  2
Affiliations
  • 1. Department of Chemical, Biological, Pharmaceutical and Environmental Sciences (CHIBIOFARAM), University of Messina, Viale F. Stagno d'Alcontres 31, 98166 Messina, Italy.
  • 2. Department of Neuroscience, Reproductive Sciences and Dentistry, University Federico II of Naples, Via S. Pansini, 5, 80131 Naples, Italy.
  • 3. Department of Pharmacy, University of Salerno, Via G. Paolo II 132, 84084 Fisciano, SA, Italy.
  • 4. Department of Science and Technology, University of Sannio, Via F. De Sanctis, 82100 Benevento, Italy.
  • 5. Department of Medicine and Health Science Vincenzo Tiberio, University of Molise, Via C. Gazzani, 86100 Campobasso, Italy.
  • 6. Department of Advanced Medical and Surgical Sciences, University of Campania "Luigi Vanvitelli", P.zza L. Miraglia 2, 80138 Naples, Italy.
  • 7. Department of Pharmacy, University Federico II of Naples, Via D. Montesano 49, 80131 Naples, Italy.
PMID: 38782404 DOI: 10.1021/acs.jmedchem.4c00268
Abstract

Gain-of-function (GoF) variants in KCNT1 channels cause severe, drug-resistant forms of epilepsy. Quinidine is a known KCNT1 blocker, but its clinical use is limited due to severe drawbacks. To identify novel KCNT1 blockers, a homology model of human KCNT1 was built and used to screen an in-house library of compounds. Among the 20 molecules selected, five (CPK4, 13, 16, 18, and 20) showed strong KCNT1-blocking ability in an in vitro fluorescence-based assay. Patch-clamp experiments confirmed a higher KCNT1-blocking potency of these compounds when compared to quinidine, and their selectivity for KCNT1 over hERG and Kv7.2 channels. Among identified molecules, CPK20 displayed the highest metabolic stability; this compound also blocked KCNT2 currents, although with a lower potency, and counteracted GoF effects prompted by 2 recurrent epilepsy-causing KCNT1 variants (G288S and A934T). The present results provide solid rational basis for future design of novel compounds to counteract KCNT1-related neurological disorders.

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