title: Discovery and Characterization of Allosteric WNK Kinase Inhibitors.
abstract: Protein kinases are known for their highly conserved adenosine triphosphate (ATP)-binding site, rendering the discovery of selective inhibitors a major challenge. In theory, allosteric inhibitors can achieve high selectivity by targeting less conserved regions of the kinases, often with an added benefit of retaining efficacy under high physiological ATP concentration. Although often overlooked in favor of ATP-site directed approaches, performing a screen at high ATP concentration or stringent hit triaging with high ATP concentration offers conceptually simple methods of identifying inhibitors that bind outside the ATP pocket. Here, we applied the latter approach to the With-No-Lysine (K) (WNK) kinases to discover lead molecules for a next-generation antihypertensive that requires a stringent safety profile. This strategy yielded several ATP noncompetitive WNK1-4 kinase inhibitors, the optimization of which enabled cocrystallization with WNK1, revealing an allosteric binding mode consistent with the observed exquisite specificity for WNK1-4 kinases. The optimized compound inhibited rubidium uptake by sodium chloride cotransporter 1 (NKCC1) in HT29 cells, consistent with the reported physiology of WNK kinases in renal electrolyte handling.
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.|Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Yamada
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Ji-Hu
authors.initials: JH
authors.lastname: Zhang
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Xie
authors.affiliation: Novartis Institutes for BioMedical Research , Novartis Pharma AG, Basel, 4002, Switzerland.
authors.forename: Juergen
authors.initials: J
authors.lastname: Reinhardt
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Amy Qiongshu
authors.initials: AQ
authors.lastname: Xie
authors.affiliation: Novartis Institutes for BioMedical Research , Novartis Pharmaceuticals Corporation, East Hanover, New Jersey 07936-1080, United States.
authors.forename: Daniel
authors.initials: D
authors.lastname: LaSala
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Darcy
authors.initials: D
authors.lastname: Kohls
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: David
authors.initials: D
authors.lastname: Yowe
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Debra
authors.initials: D
authors.lastname: Burdick
authors.affiliation: Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Yoshisue
authors.affiliation: Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Wakai
authors.affiliation: Novartis Institutes for BioMedical Research , Novartis Pharma AG, Basel, 4002, Switzerland.
authors.forename: Isabel
authors.initials: I
authors.lastname: Schmidt
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Jason
authors.initials: J
authors.lastname: Gunawan
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.|Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Kayo
authors.initials: K
authors.lastname: Yasoshima
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Q Kimberley
authors.initials: QK
authors.lastname: Yue
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.|Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Mitsunori
authors.initials: M
authors.lastname: Kato
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.|Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Muneto
authors.initials: M
authors.lastname: Mogi
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Neeraja
authors.initials: N
authors.lastname: Idamakanti
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Natasha
authors.initials: N
authors.lastname: Kreder
authors.affiliation: Novartis Institutes for BioMedical Research , Novartis Pharma AG, Basel, 4002, Switzerland.
authors.forename: Peter
authors.initials: P
authors.lastname: Drueckes
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Pramod
authors.initials: P
authors.lastname: Pandey
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.|Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Kawanami
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Waanjeng
authors.initials: W
authors.lastname: Huang
authors.affiliation: Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Yukiko I
authors.initials: YI
authors.lastname: Yagi
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.
authors.forename: Zhan
authors.initials: Z
authors.lastname: Deng
authors.affiliation: Novartis Institutes for BioMedical Research, Inc. , Cambridge, Massachusetts 02139-4133, United States.|Novartis Institutes for BioMedical Research , Novartis Pharma K.K., Tsukuba, Ibaraki 300-2611, Japan.
authors.forename: Hyi-Man
authors.initials: HM
authors.lastname: Park
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000494:Allosteric Regulation / Q000187:drug effects*; D055808:Drug Discovery; D057809:HEK293 Cells; D019073:HT29 Cells; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D015778:Minor Histocompatibility Antigens / Q000737:chemistry / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D064506:Solute Carrier Family 12, Member 2 / Q000378:metabolism; D000076204:WNK Lysine-Deficient Protein Kinase 1
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D015778:Minor Histocompatibility Antigens; D047428:Protein Kinase Inhibitors; C578877:SLC12A2 protein, human; D064506:Solute Carrier Family 12, Member 2; D000255:Adenosine Triphosphate; D017346:Protein Serine-Threonine Kinases; D000076204:WNK Lysine-Deficient Protein Kinase 1; C498743:WNK1 protein, human