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Small-molecule screen identifies inhibitors of the neuronal K-Cl cotransporter KCC2
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| Zeitschriftentitel: | Proceedings of the National Academy of Sciences |
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| Personen und Körperschaften: | , , , , , , |
| In: | Proceedings of the National Academy of Sciences, 106, 2009, 13, S. 5383-5388 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Proceedings of the National Academy of Sciences
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| Schlagwörter: |
| Zusammenfassung: | <jats:p> KCC2, a neuronal-specific K-Cl cotransporter, plays a major role in maintaining intracellular Cl <jats:sup>−</jats:sup> concentration in neurons below its electrochemical equilibrium potential, thus favoring robust GABA hyperpolarizing or inhibitory responses. The pharmacology of the K-Cl cotransporter is dominated by loop diuretics such as furosemide and bumetanide, molecules used in clinical medicine because they inhibit the loop of Henle Na-K-2Cl cotransporter with much higher affinity. To identify molecules that affect KCC2 activity, we developed a fluorescence-based assay suitable for high-throughput screening (HTS) and used the assay to screen a library of 234,000 small molecules. We identified a large number of molecules that either decrease or increase the activity of the cotransporter. Here, we report the characterization of a small number of inhibitors, some of which inhibit KCC2 activity in the submicomolar range without substantially affecting NKCC1 activity. Using medicinal chemistry, we synthesized a number of variants, tested their effect on KCC2 function, and provide an analysis of structure/activity relationships. We also used one of the compounds to demonstrate competitive inhibition in regard to external [K <jats:sup>+</jats:sup> ] versus noncompetitive inhibition in respect to external [Cl <jats:sup>−</jats:sup> ]. </jats:p> |
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| Umfang: | 5383-5388 |
| ISSN: |
0027-8424
1091-6490 |
| DOI: | 10.1073/pnas.0812756106 |