trpm8 fa  (Bio Basic Canada)

Journal: ACS chemical neuroscience

Article Title: Structure-Based Design of Novel Biphenyl Amide Antagonists of Human Transient Receptor Potential Cation Channel Subfamily M Member 8 Channels with Potential Implications in the Treatment of Sensory Neuropathies

doi: 10.1021/acschemneuro.9b00404

Figure Lengend Snippet: Reported TRPM8 antagonists. IC50 values (antagonists) determined in the Ca2+ flux assay against the effect of menthol and/or icilin: (a) ref 26, (b) ref 11, (c) ref 10, (d) ref 27, (e) ref 12, (f) ref 28, (g) ref 29, (h) ref 30, and (i) ref 31. Icilin: 3-(2-hydroxyphenyl)-6-(3-nitrophenyl)-3,4-dihydropyrimidin-2(1H)-one.

Article Snippet: In light of these recent developments in the TRPM8 field, we hypothesize that reported small molecule TRPM8 antagonists with both structural similarities and conformational rigidity can reveal similar molecular determinants for ligand recognition, via their poses from molecular dynamics (MD) simulations in our human TRPM8 homology model based on the avian cryo-EM structure TRPM8 FA (PDB 6BPQ, ~4.1 Å).

Techniques: Flux Assay

Journal: ACS chemical neuroscience

Article Title: Structure-Based Design of Novel Biphenyl Amide Antagonists of Human Transient Receptor Potential Cation Channel Subfamily M Member 8 Channels with Potential Implications in the Treatment of Sensory Neuropathies

doi: 10.1021/acschemneuro.9b00404

Figure Lengend Snippet: Superimposition of AMG2850 and (−)-menthyl 1 from their MD poses. (Green and magenta, MD pose of TRPM8 and (−)-menthyl 1; blue and gray, MD pose of TRPM8 and AMG2850).

Article Snippet: In light of these recent developments in the TRPM8 field, we hypothesize that reported small molecule TRPM8 antagonists with both structural similarities and conformational rigidity can reveal similar molecular determinants for ligand recognition, via their poses from molecular dynamics (MD) simulations in our human TRPM8 homology model based on the avian cryo-EM structure TRPM8 FA (PDB 6BPQ, ~4.1 Å).

Techniques:

Journal: ACS chemical neuroscience

Article Title: Structure-Based Design of Novel Biphenyl Amide Antagonists of Human Transient Receptor Potential Cation Channel Subfamily M Member 8 Channels with Potential Implications in the Treatment of Sensory Neuropathies

doi: 10.1021/acschemneuro.9b00404

Figure Lengend Snippet: Evaluation of test compounds for any effect on icilin-evoked Ca2+ entry signals using fura-2 based Ca2+ imaging of HEK-293 cells stably expressing human TRPM8. The fura-2 loaded cells bathed in Ca2+-free extracellular solution were exposed to icilin (500 nM) with or without the synthesized compounds or the known TRPM8 antagonist RQ-00203078 at various concentrations (For this figure, all were at 3 nM. The corresponding histogram is provided in Figure S3). After ~4 min of compound addition, Ca2+ free bath solution was replaced by the one containing 2 mM Ca2+. Ca2+ influx was monitored as fura-2 fluorescence ratio (F355/F380). Each data point represents mean ± SEM (n ≥ 35 cells from three independent experiments done in three different days).

Article Snippet: In light of these recent developments in the TRPM8 field, we hypothesize that reported small molecule TRPM8 antagonists with both structural similarities and conformational rigidity can reveal similar molecular determinants for ligand recognition, via their poses from molecular dynamics (MD) simulations in our human TRPM8 homology model based on the avian cryo-EM structure TRPM8 FA (PDB 6BPQ, ~4.1 Å).

Techniques: Imaging, Stable Transfection, Expressing, Synthesized, Fluorescence

Journal: ACS chemical neuroscience

Article Title: Structure-Based Design of Novel Biphenyl Amide Antagonists of Human Transient Receptor Potential Cation Channel Subfamily M Member 8 Channels with Potential Implications in the Treatment of Sensory Neuropathies

doi: 10.1021/acschemneuro.9b00404

Figure Lengend Snippet: Inhibition of menthol-evoked TRPM8 currents by 14. Currents were measured by whole-cell patch-clamp electrophysiology of HEK-293 cells transiently transfected with human TRPM8. (A) Average current traces (n = 4) measured by a + 80 mV voltage pulse upon exposure to a saturating concentration of 500 μM menthol and varying concentrations of 14. (B) Dose response of 14 measured at concentrations from 1 nM to 1 μM at +80 mV in the presence of 500 μM menthol. Current response was normalized to the maximum current magnitude measured without antagonist for each cell. Data was fit with a single binding site Hill equation, and the IC50 was calculated to be 64 ± 2 nM (n = 6 cells). Error bars represent standard error of the mean.

Article Snippet: In light of these recent developments in the TRPM8 field, we hypothesize that reported small molecule TRPM8 antagonists with both structural similarities and conformational rigidity can reveal similar molecular determinants for ligand recognition, via their poses from molecular dynamics (MD) simulations in our human TRPM8 homology model based on the avian cryo-EM structure TRPM8 FA (PDB 6BPQ, ~4.1 Å).

Techniques: Inhibition, Patch Clamp, Transfection, Concentration Assay, Binding Assay

Journal: ACS chemical neuroscience

Article Title: Structure-Based Design of Novel Biphenyl Amide Antagonists of Human Transient Receptor Potential Cation Channel Subfamily M Member 8 Channels with Potential Implications in the Treatment of Sensory Neuropathies

doi: 10.1021/acschemneuro.9b00404

Figure Lengend Snippet: Binding pocket of 14 in TRPM8, TRPA1, and TRPV1 and their binding mode. (a) Position of the binding pocket of 14 in TRPM8, TRPA1, and TRPV1. (b) Binding mode of 14 in hTRPA1 (PDB 3J9P). (c) Binding mode of 14 in hTRPM8. Our hTRPM8 homology model was constructed using the cryo-electron microscopy (cryo-EM) structure of TRPM8FA (PDB 6BPQ) as a template. (d) Binding mode of 14 in hTRPV1 (PDB 3J5R).

Article Snippet: In light of these recent developments in the TRPM8 field, we hypothesize that reported small molecule TRPM8 antagonists with both structural similarities and conformational rigidity can reveal similar molecular determinants for ligand recognition, via their poses from molecular dynamics (MD) simulations in our human TRPM8 homology model based on the avian cryo-EM structure TRPM8 FA (PDB 6BPQ, ~4.1 Å).

Techniques: Binding Assay, Construct, Cryo-Electron Microscopy, Cryo-EM Sample Prep