Potent Activation of Large-Conductance Ca2+-Activated K+ Channels by the Diphenylurea 1,3-Bis-[2-hydroxy-5-(trifluoromethyl)phenyl]urea (NS1643) in Pituitary Tumor (GH3) Cells

• Published in: Molecular pharmacology Vol. 74; no. 6; pp. 1696 - 1704
• Main Authors: Wu, Sheng-Nan, Peng, Hsung, Chen, Bing-Shuo, Wang, Ya-Jean, Wu, Pei-Yu, Lin, Ming-Wei
• Format: Journal Article
• Language: English
• Published: United States American Society for Pharmacology and Experimental Therapeutics 01.12.2008
• Subjects: Action Potentials; Animals; Calcium - metabolism; Cell Line, Tumor; Cresols - pharmacology; ERG1 Potassium Channel; Ether-A-Go-Go Potassium Channels - agonists; Humans; Ion Channel Gating; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits - genetics; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits - physiology; Large-Conductance Calcium-Activated Potassium Channels - agonists; Patch-Clamp Techniques; Phenylurea Compounds - pharmacology; Pituitary Neoplasms; Rats
• ISSN: 0026-895X; 1521-0111
• DOI: 10.1124/mol.108.049106

1,3-Bis-[2-hydroxy-5-(trifluoromethyl)phenyl]urea (NS1643) is reported to be an activator of human ether-à-go-go -related gene current. However, it remains unknown whether it has any effects on other types of ion channels. The effects of NS1643 on ion currents and membrane potential were investigated in this study. NS1643 stimulated Ca 2+ -activated K + current [I K(Ca) ] in a concentration-dependent manner with an EC 50 value of 1.8 μM in pituitary tumor (GH 3 ) cells. In inside-out recordings, this compound applied to the intracellular side of the detached channels stimulated large-conductance Ca 2+ -activated K + (BK Ca ) channels with no change in single-channel conductance. It shifted the activation curve of BK Ca channels to less depolarized voltages without altering the gating charge of the channels. NS1643-stimulated channel activity depended on intracellular Ca 2+ , and mean closed time during exposure to NS1643 was reduced. NS1643 (3 μM) had little or no effect on peak amplitude of ether-à-go-go -related gene-mediated K + current evoked by membrane hyperpolarization, although it increased the amplitude of late-sustained components of K + inward current, which was suppressed by paxilline but not by azimilide. NS1643 (3 μM) had no effect on L-type Ca 2+ current. This compound reduced repetitive firing of action potentials, and further application of paxilline attenuated its decrease in firing rate. In addition, NS1643 enhanced BK Ca -channel activity in human embryonic kidney 293T cells expressing α -hSlo . In summary, we clearly show that NS1643 interacts directly with the BK Ca channel to increase the amplitude of I K(Ca) in pituitary tumor (GH 3 ) cells. The α-subunit of the channel may be a target for the action of this small compound.