High-throughput screening reveals a small-molecule inhibitor of the renal outer medullary potassium channel and Kir7.1.
The renal outer medullary potassium channel (ROMK) is expressed in the kidney tubule and critically regulates sodium and potassium balance. The physiological functions of other inward rectifying K(+) (Kir) channels expressed in the nephron, such as Kir7.1, are less well understood in part due to the lack of selective pharmacological probes targeting inward rectifiers. In an effort to identify Kir channel probes, we performed a fluorescence-based, high-throughput screen (HTS) of 126,009 small molecules for modulators of ROMK function. Several antagonists were identified in the screen. One compound, termed VU590, inhibits ROMK with submicromolar affinity, but has no effect on Kir2.1 or Kir4.1. Low micromolar concentrations inhibit Kir7.1, making VU590 the first small-molecule inhibitor of Kir7.1. Structure-activity relationships of VU590 were defined using small-scale parallel synthesis. Electrophysiological analysis indicates that VU590 is an intracellular pore blocker. VU590 and other compounds identified by HTS will be instrumental in defining Kir channel structure, physiology, and therapeutic potential.
Lewis
L Michelle
LM
Vanderbilt Institute of Chemical Biology, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
Bhave
Gautam
G
Chauder
Brian A
BA
Banerjee
Sreedatta
S
Lornsen
Katharina A
KA
Redha
Rey
R
Fallen
Katherine
K
Lindsley
Craig W
CW
Weaver
C David
CD
Denton
Jerod S
JS
D000818:Animals; D002460:Cell Line; D005820:Genetic Testing / Q000379:methods*; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry* / Q000494:pharmacology*; D006801:Humans; D007679:Kidney Medulla / Q000187:drug effects / Q000502:physiology*; D026902:Potassium Channel Blockers / Q000737:chemistry / Q000494:pharmacology*; D024661:Potassium Channels, Inwardly Rectifying / Q000037:antagonists & inhibitors* / Q000502:physiology*; D051381:Rats; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*
C544855:7,13-bis(4-nitrobenzyl)-1,4,10-trioxa-7,13-diazacyclopentadecane; D006573:Heterocyclic Compounds, 1-Ring; C489477:Kir7.1 channel; D026902:Potassium Channel Blockers; D024661:Potassium Channels, Inwardly Rectifying; D054852:Small Molecule Libraries
J Biomol Screen. 1999;4(2):67-73
10838414
Am J Physiol Renal Physiol. 2007 Jul;293(1):F227-35
17389680
J Biol Chem. 2003 Jun 20;278(25):23066-75
12684516
J Biol Chem. 2002 Oct 4;277(40):37871-80
12122007
J Biomol Screen. 2004 Dec;9(8):671-7
15634793
J Am Coll Cardiol. 2004 Jan 21;43(2):155-61
14736430
Physiol Rev. 2005 Jan;85(1):319-71
15618483
Am J Physiol Renal Physiol. 2008 Jun;294(6):F1398-407
18367659
Nature. 1994 Sep 15;371(6494):243-6
7915826
Channels (Austin). 2007 May-Jun;1(3):172-8
18690029
J Neurophysiol. 2000 Aug;84(2):1062-75
10938328
Hypertension. 2008 Jun;51(6):1658-64
18443236
J Membr Biol. 1984;80(3):221-33
6094817
Exp Eye Res. 2008 Sep;87(3):176-83
18653180
Nature. 1994 Nov 24;372(6504):366-9
7969496
Mol Pharmacol. 2009 Jun;75(6):1287-95
19264848
J Biol Chem. 2005 Oct 21;280(42):35281-9
16118216
J Am Soc Nephrol. 2000 Nov;11(11):1987-1994
11053473
Am J Hum Genet. 2008 Jan;82(1):174-80
18179896
Nat Genet. 2008 May;40(5):592-599
18391953
Channels (Austin). 2009 Jan-Feb;3(1):57-68
19221509
Pflugers Arch. 2009 May;458(1):157-68
18839206
Nature. 1994 Sep 15;371(6494):246-9
8078584
Proc Natl Acad Sci U S A. 2008 Jan 29;105(4):1364-8
18216262
J Membr Biol. 1984;80(3):201-19
6094816
Biochem Biophys Res Commun. 1999 Aug 2;261(2):364-71
10425191
J Gen Physiol. 2005 Apr;125(4):413-26
15795311
Nat Genet. 1996 Oct;14(2):152-6
8841184
Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):15298-303
10611379
Am J Physiol. 1998 Sep;275(3):F325-7
9729502
Proc Natl Acad Sci U S A. 2005 Jul 12;102(28):9954-9
15987778
Hum Mutat. 2006 Apr;27(4):388
16541386
Am J Physiol Renal Physiol. 2009 Feb;296(2):F347-54
19036846
Neuron. 1998 May;20(5):995-1005
9620703
Cell. 1995 Jan 13;80(1):149-54
7813010
Am J Physiol. 1997 Nov;273(5):F825-36
9374848
Nat Biotechnol. 2006 Feb;24(2):167-75
16465162
J Biol Chem. 2002 Oct 4;277(40):37881-7
12130653
J Hypertens. 1995 Dec;13(12 Pt 2):1539-45
8903607
Proc Natl Acad Sci U S A. 2009 Mar 31;106(13):5383-8
19279215