Discovery, characterization, and effects on renal fluid and electrolyte excretion of the Kir4.1 potassium channel pore blocker, VU0134992

Sujay V. Kharade, Haruto Kurata, Aaron M. Bender, Anna L. Blobaum, Eric E. Figueroa, Amanda Duran, Meghan Kramer, Emily Days, Paige Vinson, Daniel Flores, Lisa M. Satlin, Jens Meiler, C. David Weaver, Craig W. Lindsley, Corey R Hopkins, Jerod S. Denton

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The inward rectifier potassium (Kir) channel Kir4.1 (KCNJ10) carries out important physiologic roles in epithelial cells of the kidney, astrocytes in the central nervous system, and stria vascularis of the inner ear. Loss-of-function mutations in KCNJ10 lead to EAST/SeSAME syndrome, which is characterized by epilepsy, ataxia, renal salt wasting, and sensorineural deafness. Although genetic approaches have been indispensable for establishing the importance of Kir4.1 in the normal function of these tissues, the availability of pharmacological tools for acutely manipulating the activity of Kir4.1 in genetically normal animals has been lacking. We therefore carried out a high-throughput screen of 76, 575 compounds from the Vanderbilt Institute of Chemical Biology library for small-molecule modulators of Kir4.1. The most potent inhibitor identified was 2-(2-bromo-4-isopropylphenoxy)- N-(2, 2, 6, 6-tetramethylpiperidin-4-yl)acetamide (VU0134992). In whole-cell patch-clamp electrophysiology experiments, VU0134992 inhibits Kir4.1 with an IC 50 value of 0.97 μM and is 9-fold selective for homomeric Kir4.1 over Kir4.1/5.1 concatemeric channels (IC 50 5 9 μM) at 2120 mV. In thallium (Tl + ) flux assays, VU0134992 is greater than 30-fold selective for Kir4.1 over Kir1.1, Kir2.1, and Kir2.2; is weakly active toward Kir2.3, Kir6.2/SUR1, and Kir7.1; and is equally active toward Kir3.1/3.2, Kir3.1/3.4, and Kir4.2. This potency and selectivity profile is superior to Kir4.1 inhibitors amitriptyline, nortriptyline, and fluoxetine. Medicinal chemistry identified components of VU0134992 that are critical for inhibiting Kir4.1. Patch-clamp electrophysiology, molecular modeling, and site-directed mutagenesis identified pore-lining glutamate 158 and isoleucine 159 as critical residues for block of the channel. VU0134992 displayed a large free unbound fraction (f u ) in rat plasma (f u 5 0.213). Consistent with the known role of Kir4.1 in renal function, oral dosing of VU0134992 led to a dose-dependent diuresis, natriuresis, and kaliuresis in rats. Thus, VU0134992 represents the first in vivo active tool compound for probing the therapeutic potential of Kir4.1 as a novel diuretic target for the treatment of hypertension.

Original languageEnglish (US)
Pages (from-to)926-937
Number of pages12
JournalMolecular pharmacology
Volume94
Issue number2
DOIs
StatePublished - Aug 1 2018

Fingerprint

Potassium Channel Blockers
Electrolytes
Electrophysiology
Kidney
Stria Vascularis
Small Molecule Libraries
Inwardly Rectifying Potassium Channel
Nortriptyline
Natriuresis
Amitriptyline
Pharmaceutical Chemistry
Isoleucine
Thallium
Fluoxetine
Diuresis
Deafness
Inner Ear
Ataxia
Site-Directed Mutagenesis
Diuretics

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

Cite this

Kharade, S. V., Kurata, H., Bender, A. M., Blobaum, A. L., Figueroa, E. E., Duran, A., ... Denton, J. S. (2018). Discovery, characterization, and effects on renal fluid and electrolyte excretion of the Kir4.1 potassium channel pore blocker, VU0134992. Molecular pharmacology, 94(2), 926-937. https://doi.org/10.1124/mol.118.112359

Discovery, characterization, and effects on renal fluid and electrolyte excretion of the Kir4.1 potassium channel pore blocker, VU0134992. / Kharade, Sujay V.; Kurata, Haruto; Bender, Aaron M.; Blobaum, Anna L.; Figueroa, Eric E.; Duran, Amanda; Kramer, Meghan; Days, Emily; Vinson, Paige; Flores, Daniel; Satlin, Lisa M.; Meiler, Jens; Weaver, C. David; Lindsley, Craig W.; Hopkins, Corey R; Denton, Jerod S.

In: Molecular pharmacology, Vol. 94, No. 2, 01.08.2018, p. 926-937.

Research output: Contribution to journalArticle

Kharade, SV, Kurata, H, Bender, AM, Blobaum, AL, Figueroa, EE, Duran, A, Kramer, M, Days, E, Vinson, P, Flores, D, Satlin, LM, Meiler, J, Weaver, CD, Lindsley, CW, Hopkins, CR & Denton, JS 2018, 'Discovery, characterization, and effects on renal fluid and electrolyte excretion of the Kir4.1 potassium channel pore blocker, VU0134992', Molecular pharmacology, vol. 94, no. 2, pp. 926-937. https://doi.org/10.1124/mol.118.112359
Kharade, Sujay V. ; Kurata, Haruto ; Bender, Aaron M. ; Blobaum, Anna L. ; Figueroa, Eric E. ; Duran, Amanda ; Kramer, Meghan ; Days, Emily ; Vinson, Paige ; Flores, Daniel ; Satlin, Lisa M. ; Meiler, Jens ; Weaver, C. David ; Lindsley, Craig W. ; Hopkins, Corey R ; Denton, Jerod S. / Discovery, characterization, and effects on renal fluid and electrolyte excretion of the Kir4.1 potassium channel pore blocker, VU0134992. In: Molecular pharmacology. 2018 ; Vol. 94, No. 2. pp. 926-937.
@article{c7fa784623774dc3bc164ae0d57c8c80,
title = "Discovery, characterization, and effects on renal fluid and electrolyte excretion of the Kir4.1 potassium channel pore blocker, VU0134992",
abstract = "The inward rectifier potassium (Kir) channel Kir4.1 (KCNJ10) carries out important physiologic roles in epithelial cells of the kidney, astrocytes in the central nervous system, and stria vascularis of the inner ear. Loss-of-function mutations in KCNJ10 lead to EAST/SeSAME syndrome, which is characterized by epilepsy, ataxia, renal salt wasting, and sensorineural deafness. Although genetic approaches have been indispensable for establishing the importance of Kir4.1 in the normal function of these tissues, the availability of pharmacological tools for acutely manipulating the activity of Kir4.1 in genetically normal animals has been lacking. We therefore carried out a high-throughput screen of 76, 575 compounds from the Vanderbilt Institute of Chemical Biology library for small-molecule modulators of Kir4.1. The most potent inhibitor identified was 2-(2-bromo-4-isopropylphenoxy)- N-(2, 2, 6, 6-tetramethylpiperidin-4-yl)acetamide (VU0134992). In whole-cell patch-clamp electrophysiology experiments, VU0134992 inhibits Kir4.1 with an IC 50 value of 0.97 μM and is 9-fold selective for homomeric Kir4.1 over Kir4.1/5.1 concatemeric channels (IC 50 5 9 μM) at 2120 mV. In thallium (Tl + ) flux assays, VU0134992 is greater than 30-fold selective for Kir4.1 over Kir1.1, Kir2.1, and Kir2.2; is weakly active toward Kir2.3, Kir6.2/SUR1, and Kir7.1; and is equally active toward Kir3.1/3.2, Kir3.1/3.4, and Kir4.2. This potency and selectivity profile is superior to Kir4.1 inhibitors amitriptyline, nortriptyline, and fluoxetine. Medicinal chemistry identified components of VU0134992 that are critical for inhibiting Kir4.1. Patch-clamp electrophysiology, molecular modeling, and site-directed mutagenesis identified pore-lining glutamate 158 and isoleucine 159 as critical residues for block of the channel. VU0134992 displayed a large free unbound fraction (f u ) in rat plasma (f u 5 0.213). Consistent with the known role of Kir4.1 in renal function, oral dosing of VU0134992 led to a dose-dependent diuresis, natriuresis, and kaliuresis in rats. Thus, VU0134992 represents the first in vivo active tool compound for probing the therapeutic potential of Kir4.1 as a novel diuretic target for the treatment of hypertension.",
author = "Kharade, {Sujay V.} and Haruto Kurata and Bender, {Aaron M.} and Blobaum, {Anna L.} and Figueroa, {Eric E.} and Amanda Duran and Meghan Kramer and Emily Days and Paige Vinson and Daniel Flores and Satlin, {Lisa M.} and Jens Meiler and Weaver, {C. David} and Lindsley, {Craig W.} and Hopkins, {Corey R} and Denton, {Jerod S.}",
year = "2018",
month = "8",
day = "1",
doi = "10.1124/mol.118.112359",
language = "English (US)",
volume = "94",
pages = "926--937",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "2",

}

TY - JOUR

T1 - Discovery, characterization, and effects on renal fluid and electrolyte excretion of the Kir4.1 potassium channel pore blocker, VU0134992

AU - Kharade, Sujay V.

AU - Kurata, Haruto

AU - Bender, Aaron M.

AU - Blobaum, Anna L.

AU - Figueroa, Eric E.

AU - Duran, Amanda

AU - Kramer, Meghan

AU - Days, Emily

AU - Vinson, Paige

AU - Flores, Daniel

AU - Satlin, Lisa M.

AU - Meiler, Jens

AU - Weaver, C. David

AU - Lindsley, Craig W.

AU - Hopkins, Corey R

AU - Denton, Jerod S.

PY - 2018/8/1

Y1 - 2018/8/1

N2 - The inward rectifier potassium (Kir) channel Kir4.1 (KCNJ10) carries out important physiologic roles in epithelial cells of the kidney, astrocytes in the central nervous system, and stria vascularis of the inner ear. Loss-of-function mutations in KCNJ10 lead to EAST/SeSAME syndrome, which is characterized by epilepsy, ataxia, renal salt wasting, and sensorineural deafness. Although genetic approaches have been indispensable for establishing the importance of Kir4.1 in the normal function of these tissues, the availability of pharmacological tools for acutely manipulating the activity of Kir4.1 in genetically normal animals has been lacking. We therefore carried out a high-throughput screen of 76, 575 compounds from the Vanderbilt Institute of Chemical Biology library for small-molecule modulators of Kir4.1. The most potent inhibitor identified was 2-(2-bromo-4-isopropylphenoxy)- N-(2, 2, 6, 6-tetramethylpiperidin-4-yl)acetamide (VU0134992). In whole-cell patch-clamp electrophysiology experiments, VU0134992 inhibits Kir4.1 with an IC 50 value of 0.97 μM and is 9-fold selective for homomeric Kir4.1 over Kir4.1/5.1 concatemeric channels (IC 50 5 9 μM) at 2120 mV. In thallium (Tl + ) flux assays, VU0134992 is greater than 30-fold selective for Kir4.1 over Kir1.1, Kir2.1, and Kir2.2; is weakly active toward Kir2.3, Kir6.2/SUR1, and Kir7.1; and is equally active toward Kir3.1/3.2, Kir3.1/3.4, and Kir4.2. This potency and selectivity profile is superior to Kir4.1 inhibitors amitriptyline, nortriptyline, and fluoxetine. Medicinal chemistry identified components of VU0134992 that are critical for inhibiting Kir4.1. Patch-clamp electrophysiology, molecular modeling, and site-directed mutagenesis identified pore-lining glutamate 158 and isoleucine 159 as critical residues for block of the channel. VU0134992 displayed a large free unbound fraction (f u ) in rat plasma (f u 5 0.213). Consistent with the known role of Kir4.1 in renal function, oral dosing of VU0134992 led to a dose-dependent diuresis, natriuresis, and kaliuresis in rats. Thus, VU0134992 represents the first in vivo active tool compound for probing the therapeutic potential of Kir4.1 as a novel diuretic target for the treatment of hypertension.

AB - The inward rectifier potassium (Kir) channel Kir4.1 (KCNJ10) carries out important physiologic roles in epithelial cells of the kidney, astrocytes in the central nervous system, and stria vascularis of the inner ear. Loss-of-function mutations in KCNJ10 lead to EAST/SeSAME syndrome, which is characterized by epilepsy, ataxia, renal salt wasting, and sensorineural deafness. Although genetic approaches have been indispensable for establishing the importance of Kir4.1 in the normal function of these tissues, the availability of pharmacological tools for acutely manipulating the activity of Kir4.1 in genetically normal animals has been lacking. We therefore carried out a high-throughput screen of 76, 575 compounds from the Vanderbilt Institute of Chemical Biology library for small-molecule modulators of Kir4.1. The most potent inhibitor identified was 2-(2-bromo-4-isopropylphenoxy)- N-(2, 2, 6, 6-tetramethylpiperidin-4-yl)acetamide (VU0134992). In whole-cell patch-clamp electrophysiology experiments, VU0134992 inhibits Kir4.1 with an IC 50 value of 0.97 μM and is 9-fold selective for homomeric Kir4.1 over Kir4.1/5.1 concatemeric channels (IC 50 5 9 μM) at 2120 mV. In thallium (Tl + ) flux assays, VU0134992 is greater than 30-fold selective for Kir4.1 over Kir1.1, Kir2.1, and Kir2.2; is weakly active toward Kir2.3, Kir6.2/SUR1, and Kir7.1; and is equally active toward Kir3.1/3.2, Kir3.1/3.4, and Kir4.2. This potency and selectivity profile is superior to Kir4.1 inhibitors amitriptyline, nortriptyline, and fluoxetine. Medicinal chemistry identified components of VU0134992 that are critical for inhibiting Kir4.1. Patch-clamp electrophysiology, molecular modeling, and site-directed mutagenesis identified pore-lining glutamate 158 and isoleucine 159 as critical residues for block of the channel. VU0134992 displayed a large free unbound fraction (f u ) in rat plasma (f u 5 0.213). Consistent with the known role of Kir4.1 in renal function, oral dosing of VU0134992 led to a dose-dependent diuresis, natriuresis, and kaliuresis in rats. Thus, VU0134992 represents the first in vivo active tool compound for probing the therapeutic potential of Kir4.1 as a novel diuretic target for the treatment of hypertension.

UR - http://www.scopus.com/inward/record.url?scp=85050771117&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85050771117&partnerID=8YFLogxK

U2 - 10.1124/mol.118.112359

DO - 10.1124/mol.118.112359

M3 - Article

VL - 94

SP - 926

EP - 937

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 2

ER -