Bookmark and Share
BioAssay: AID 2262

Summary of probe development for inhibitors of KCNQ2 potassium channel

Assay Implementation: Haibo Yu Ph.D., Beiyan Zou Ph.D., Shunyou Long M.S., Amy Scott, Meng Wu Ph.D., Joseph Babcock, Bill Shi Ph.D., David Meyers Ph.D., Jia Xu Ph.D. ..more
_
   
AID: 2262
Data Source: Johns Hopkins Ion Channel Center (JHICC_KCNQ2_potentiator_Summary)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-01-15
Modify Date: 2013-03-27
Target
Sequence: potassium voltage-gated channel KQT-like protein 2 [Rattus norvegicus]
Description ..   
Protein Family: KCNQ voltage-gated potassium channel

Gene:KCNQ2     Related Protein 3D Structures     More BioActivity Data..
Depositor Specified Assays
Show more
AIDNameTypeComment
2156Primary cell-based high-throughput screening assay for identification of compounds that inhibit KCNQ2 potassium channelsscreeningPrimary HTS assay for KCNQ2 inhibitors where 305615 compounds are screened and 3405 are found active.
504839Dose response assay for compounds that inhibit KCNQ2 potassium channels on automated electrophysiological assayconfirmatoryDose response assay for compounds that inhibit KCNQ2 potassium channels on automated electrophysiological assay
504837SAR analysis for compounds that inhibit KCNQ2 potassium channels on automated electrophysiological assayconfirmatorySAR analysis for compounds that inhibit KCNQ2 potassium channels on automated electrophysiological assay
493025Confirmatory screen for compounds that inhibit KCNQ2 potassium channelsscreening
493026Specificity screen against KCNQ1 for compounds that inhibit KCNQ2 potassium channelsscreening
493029Counter screen against parental CHO cells for compounds that inhibit KCNQ2 potassium channelsscreening
588425Dose response assay of SAR compounds for the identification of selective inhibitors of KCNQ2 potassium channels in the KCNQ1 expressing cells on automated patch clampconfirmatory
588426SAR analysis for compounds that inhibit KCNQ2 potassium channels on automated electrophysiological assay, CRC2confirmatory
588531Confirmatory screen for compounds that inhibit KCNQ2 potassium channels using automated patch clampother
588635SAR-Confirmatory assay to confirm a potent KCNQ2 inhibitor using manual electrophysiologyother
588637Dose response assay for compounds that inhibit KCNQ2 potassium channels on automated electrophysiological assay IIconfirmatory
602271SAR analysis for compounds that inhibit KCNQ2 potassium channels in an automated electrophysiological assayconfirmatory
Description:
Source (MLPCN Center Name): Johns Hopkins Ion Channel Center (JHICC)
Center Affiliation: Johns Hopkins University, School of Medicine
Screening Center PI: Min Li, Ph.D.
Assay Provider: Min Li, Ph.D.
Network: Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R03 DA027716-01
Grant Proposal PI: Min Li, Ph.D., Johns Hopkins University School of Medicine
Assay Implementation: Haibo Yu Ph.D., Beiyan Zou Ph.D., Shunyou Long M.S., Amy Scott, Meng Wu Ph.D., Joseph Babcock, Bill Shi Ph.D., David Meyers Ph.D., Jia Xu Ph.D.

Description:

Voltage-gated potassium (K) channels are critical for neuronal function in excitable tissues such as brain and heart. They are also found in non-excitable tissues important for other functions such as hormone secretion, oxygen-sensing and immune responses. There are more than 100 genes in human genome encoding different but homologous potassium channels. Voltage-gated K+ channels, exemplified by the Shaker K+ channel, share considerable sequence similarity. Isolation and characterization of bioactive chemical probes could form important pharmacological foundation, providing a great deal of insights into the structure and function.

The KCNQ (or also called Kv7) channel family includes five members, KCNQ1 to KCNQ5; KCNQ are members of Kv channel superfamily[1-6]. Different from other Kv channel classes, KCNQ channels commonly display a sub-threshold opening (i.e., at a voltage near resting potential) and are regulated by GPCR signaling, e.g., muscarinic receptors [7, 8]. KCNQ2-5 channels are primarily expressed in the brain, while KCNQ1 is abundantly expressed in cardiac tissue. Among congenital long QT patients, a disease which affects 1 in 2,500 live births, the most prevalent mutations are within the KCNQ1 allele. Mutations in KCNQ2 and KCNQ3 are known to cause benign familial neonatal convulsion (BFNC), a rare form of epilepsy [9].

In the mid-90s, a compound known as D-23129, or retigabine, was initially developed to target GABAnergic signaling [10, 11]. Despite its poor effect on the GABA pathway, retigabine was shown to have potent anti-convulsant activity. In 1997, retigabine was also found to induce the opening of voltage-gated potassium channels at resting membrane potentials, and since then, through a variety of other studies, this has been established to be its main mechanism of action. Retigabine, acting on KCNQ2/3 potassium channels which code M-currents, is now being used for treatment of epilepsy. None of the anti-convulsants that are in clinical use today have a comparable mechanism of action, i.e., through ligand activation of voltage-gated potassium channels. Several potentiation compounds are synthesized according to the scaffold of retigabine. However, considering the poor potency of retigabine and several other undesirable characteristics including its broad action on KCNQ2, 3, 4, and 5 [12], it is not clear whether these related structures will fundamentally improve therapeutic efficacy. Hence, it argues for the performance of a non-biased screen for new structures with potentiation activity.

This assay summarizes the efforts of identifying and developing small chemical compounds that inhibit the KCNQ2 potassium channels.

References:
1. Charlier, C. et al. A pore mutation in a novel KQT-like potassium channel gene in an idiopathic epilepsy family. Nat. Genet. 1998.?18, 53-55, PMID: 9425900
2. Gutman, G.A. et al. International Union of Pharmacology. XLI. Compendium of voltage-gated ion channels: potassium channels. Pharmacol. Rev. 2003. 55, 583-586, PMID: 14657415
3. Kubisch, C. et al. KCNQ4, a novel potassium channel expressed in sensory outer hair cells, is mutated in dominant deafness. Cell. 1999. 96, 437-446, PMID: 10025409
4. Schroeder, B.C., Hechenberger, M., Weinreich, F., Kubisch, C. & Jentsch, T.J. KCNQ5, a novel potassium channel broadly expressed in brain, mediates M-type currents. J. Biol. Chem. 2000, 275, 24089-24095,PMID: 10816588.
5. Singh, N.A. et al. A novel potassium channel gene, KCNQ2, is mutated in an inherited epilepsy of newborns. Nat. Genet. 1998.18, 25-29, PMID: 9425895.
6. Wang, Q. et al. Positional cloning of a novel potassium channel gene: KVLQT1 mutations cause cardiac arrhythmias. Nat. Genet. 1996, 12, 17-23, PMID: 8528244.
7. Brown, D.A. et al. Muscarinic suppression of a novel voltage-sensitive K+ current in a vertebrate neurone. Nature. 1980. 283, 673-676. PMID: 6965523.
8. Marrion, N.V. Control of M-current. Annu. Rev. Physiol.1997.59, 483-504, PMID: 9074774.
9. Maljevic, S. et al. Nervous system KV7 disorders: breakdown of a subthreshold brake. J Physiol. 2008. 586, 1791-1801. PMID: 18238816.
10. Rostock, A. et al. D-23129: a new anticonvulsant with a broad spectrum activity in animal models of epileptic seizures. Epilepsy Res. 1996. 23, 211-223. PMID: 8739124
11. Tober, C., Rostock, A., Rundfeldt, C. & Bartsch, R. D-23129: a potent anticonvulsant in the amygdala kindling model of complex partial seizures. Eur. J. Pharmacol. 1996. 303, 163-169. PMID: 8813562
12. Gribkoff, V.K. (2003) The therapeutic potential of neuronal KCNQ channel modulators. Expert Opin. Ther. Targets. 2003. 7, 737-748. PMID: 14640909.
Protocol
Please see the related bioassay (AID 2156) for details of the assay protocol.
Comment
Result of this screening campaign has yielded a probe compound, ML252.
Additional Information
Grant Number: 1 R03 DA027716-01

PageFrom: