human p2x3 receptor  (Biomol GmbH)

Journal: British Journal of Pharmacology

Article Title: Action of MK‐7264 (gefapixant) at human P2X3 and P2X2/3 receptors and in vivo efficacy in models of sensitisation

doi: 10.1111/bph.14677

Figure Lengend Snippet: MK‐7264 antagonism of human P2X3 receptor is associated with an onset of slowed desensitisation. (a) Representative trace showing a diminished peak response and appearance of slow desensitisation are associated with MK‐7264 antagonism. (b) Representative superimposed trace showing time‐dependent recovery of P2X3 fast desensitisation following MK‐7264 (300 nM) wash‐off. Agonist applied every 4 min. (c) Relationship between MK‐7264 concentration and the rate of desensitisation. Time constants (τ values) were derived from single exponential curves fitted to the desensitisation phase 250–750 ms after the peak response. The τ values were compared with a Kruskal–Wallis ANOVA (P < .05) followed by Dunn's post hoc test; N = 6. (d) The change in shape of the current following MK‐7264 antagonism quantified as a fractional response (residual current, 1 s after onset of agonist response over peak current) and compared with a Kruskal–Wallis ANOVA followed by Dunn's post hoc test. *P < 0.05 versus control response; N = 7

Article Snippet: Cells 1321N1 astrocytoma lines (RRID:CVCL_0110) stably transfected with human P2X2, P2X3, or co‐expressing P2X2 and P2X3 receptors were used in all experiments and provided by Afferent Pharmaceuticals Ltd.

Techniques: Concentration Assay, Derivative Assay, Control

Journal: British Journal of Pharmacology

Article Title: Action of MK‐7264 (gefapixant) at human P2X3 and P2X2/3 receptors and in vivo efficacy in models of sensitisation

doi: 10.1111/bph.14677

Figure Lengend Snippet: MK‐7264 activity at human P2X2, P2X3, and P2X2/3 receptors. (a–c) Representative concatenated traces of whole‐cell patch‐clamp recordings for human P2X3 (a), P2X2/3 (b), and P2X2 (c) receptor currents in 1312N1 stably transfected cells. Fully recovered P2X3 currents were evoked by 10‐μM α,β‐meATP applied for 1 s every 4 min and 10‐μM α,β‐meATP applied for 2 s at 30‐s intervals for P2X2/3 currents. P2X2 currents were evoked by 10‐μM ATP every 4 min. MK‐7264 was applied 4 min, 30 s, and 4 min prior to agonist application for P2X3 (1 μM), P2X2/3 (1 μM), and P2X2 (10 μM) receptors, respectively. Agonist application is indicated by the perfusion bar. Currents shaded in grey are in the presence of MK‐7264. (d) MK‐7264 concentration–dependent inhibition curve at human P2X3 (closed circles) and P2X2/3 receptors (open circles; N = 5). Data sets were compared with an F test and found to be significantly different from one another (F = 11.74, P = .018). (e) Effect of MK‐7264 on α,β‐meATP concentration–response curve at P2X3 receptors. Currents are in the presence of 300‐nM MK‐7264 (open circles) or vehicle control (closed circles; N = 6). MK‐7264 was applied for 4 min prior to agonist application. EC50 values were compared with a paired sample t test and found to be significantly different (P = .018). The holding potential was −80 mV for all experiments

Article Snippet: Cells 1321N1 astrocytoma lines (RRID:CVCL_0110) stably transfected with human P2X2, P2X3, or co‐expressing P2X2 and P2X3 receptors were used in all experiments and provided by Afferent Pharmaceuticals Ltd.

Techniques: Activity Assay, Patch Clamp, Stable Transfection, Transfection, Concentration Assay, Inhibition, Control

Journal: British Journal of Pharmacology

Article Title: Action of MK‐7264 (gefapixant) at human P2X3 and P2X2/3 receptors and in vivo efficacy in models of sensitisation

doi: 10.1111/bph.14677

Figure Lengend Snippet: MK‐7264 antagonism of human P2X3 receptor is associated with an onset of slowed desensitisation. (a) Representative trace showing a diminished peak response and appearance of slow desensitisation are associated with MK‐7264 antagonism. (b) Representative superimposed trace showing time‐dependent recovery of P2X3 fast desensitisation following MK‐7264 (300 nM) wash‐off. Agonist applied every 4 min. (c) Relationship between MK‐7264 concentration and the rate of desensitisation. Time constants (τ values) were derived from single exponential curves fitted to the desensitisation phase 250–750 ms after the peak response. The τ values were compared with a Kruskal–Wallis ANOVA (P < .05) followed by Dunn's post hoc test; N = 6. (d) The change in shape of the current following MK‐7264 antagonism quantified as a fractional response (residual current, 1 s after onset of agonist response over peak current) and compared with a Kruskal–Wallis ANOVA followed by Dunn's post hoc test. *P < 0.05 versus control response; N = 7

Article Snippet: 1321N1 astrocytoma lines (RRID:CVCL_0110) stably transfected with human P2X2, P2X3, or co‐expressing P2X2 and P2X3 receptors were used in all experiments and provided by Afferent Pharmaceuticals Ltd.

Techniques: Concentration Assay, Derivative Assay, Control

Journal: British Journal of Pharmacology

Article Title: Action of MK‐7264 (gefapixant) at human P2X3 and P2X2/3 receptors and in vivo efficacy in models of sensitisation

doi: 10.1111/bph.14677

Figure Lengend Snippet: MK‐7264 activity at human P2X2, P2X3, and P2X2/3 receptors. (a–c) Representative concatenated traces of whole‐cell patch‐clamp recordings for human P2X3 (a), P2X2/3 (b), and P2X2 (c) receptor currents in 1312N1 stably transfected cells. Fully recovered P2X3 currents were evoked by 10‐μM α,β‐meATP applied for 1 s every 4 min and 10‐μM α,β‐meATP applied for 2 s at 30‐s intervals for P2X2/3 currents. P2X2 currents were evoked by 10‐μM ATP every 4 min. MK‐7264 was applied 4 min, 30 s, and 4 min prior to agonist application for P2X3 (1 μM), P2X2/3 (1 μM), and P2X2 (10 μM) receptors, respectively. Agonist application is indicated by the perfusion bar. Currents shaded in grey are in the presence of MK‐7264. (d) MK‐7264 concentration–dependent inhibition curve at human P2X3 (closed circles) and P2X2/3 receptors (open circles; N = 5). Data sets were compared with an F test and found to be significantly different from one another (F = 11.74, P = .018). (e) Effect of MK‐7264 on α,β‐meATP concentration–response curve at P2X3 receptors. Currents are in the presence of 300‐nM MK‐7264 (open circles) or vehicle control (closed circles; N = 6). MK‐7264 was applied for 4 min prior to agonist application. EC50 values were compared with a paired sample t test and found to be significantly different (P = .018). The holding potential was −80 mV for all experiments

Article Snippet: 1321N1 astrocytoma lines (RRID:CVCL_0110) stably transfected with human P2X2, P2X3, or co‐expressing P2X2 and P2X3 receptors were used in all experiments and provided by Afferent Pharmaceuticals Ltd.

Techniques: Activity Assay, Patch Clamp, Stable Transfection, Transfection, Concentration Assay, Inhibition, Control