Trichloro(sulfanyl)ethyl benzamides inhibit cold temperature (4°C) induced Cauptake in CHO cells stably expressing human TRPA1

<p><b>Copyright information:</b></p><p>Taken from "Species-specific pharmacology of Trichloro(sulfanyl)ethyl benzamides as transient receptor potential ankyrin 1 (TRPA1) antagonists"</p><p>http://www.molecularpain.com/content/3/1/39</p><p>Molecular Pain 2007;3():39-39.</p><p>Published online 17 Dec 2007</p><p>PMCID:PMC2222611.</p><p></p> Concentration-response curves were generated utilizing Cauptake assays as described under . Each point in the graph is an average ± SEM of an experiment conducted in duplicate

Trichloro(sulfanyl)ethyl benzamides inhibit AITC (80 μM) induced increase in intracellular calcium and inward currents in CHO cells stably expressing human TRPA1

<p><b>Copyright information:</b></p><p>Taken from "Species-specific pharmacology of Trichloro(sulfanyl)ethyl benzamides as transient receptor potential ankyrin 1 (TRPA1) antagonists"</p><p>http://www.molecularpain.com/content/3/1/39</p><p>Molecular Pain 2007;3():39-39.</p><p>Published online 17 Dec 2007</p><p>PMCID:PMC2222611.</p><p></p> Effect of AMG9090 , AMG5445 , AMG2504 , and AMG7160 on AITC-induced increase in intracellular calcium in CHO cells expressing human TRPA1 measured in an aequorin-readout assay. Open circles represent the response of cells to the compound itself in the absence of agonist. Each point in the graph is an average ± SEM of an experiment conducted in triplicate

Effect of human TRPA1 antagonists on CHO cells stably expressing rat TRPA1

<p><b>Copyright information:</b></p><p>Taken from "Species-specific pharmacology of Trichloro(sulfanyl)ethyl benzamides as transient receptor potential ankyrin 1 (TRPA1) antagonists"</p><p>http://www.molecularpain.com/content/3/1/39</p><p>Molecular Pain 2007;3():39-39.</p><p>Published online 17 Dec 2007</p><p>PMCID:PMC2222611.</p><p></p> Concentration-response curves generated in aequorin-readout assay Note, AMG9090 and AMG5445 acted as partial agonists. Each point in the graph is presented as percent of AITC response and is an average ± SEM of an experiment conducted in triplicate. Representative traces of inward currents induced by AITC, AMG9090 and AMG5445 are shown in , , and respectively. AITC-induced increase in intracellular calcium in CHO cells expressing rat TRPA1 in the absence (100%) or presence of different concentrations of AMG2504 and AMG 7160. Representative current traces induced by AITC in the presence of AMG2504 and AMG 7160 are shown in and , respectively

Characterization of CHO cell lines stably expressing human and rat TRPA1

<p><b>Copyright information:</b></p><p>Taken from "Species-specific pharmacology of Trichloro(sulfanyl)ethyl benzamides as transient receptor potential ankyrin 1 (TRPA1) antagonists"</p><p>http://www.molecularpain.com/content/3/1/39</p><p>Molecular Pain 2007;3():39-39.</p><p>Published online 17 Dec 2007</p><p>PMCID:PMC2222611.</p><p></p> FLASH luminometer was used to measure concentration-dependent AITC-induced increase in intracellular calcium, and concentration-dependent inhibition of AITC (80 μM) activation by ruthenium red in CHO cells expressing human and rat TRPA1. Each point in the graph is an average ± SEM of an experiment conducted in triplicate. Maximum response of AITC (80 μM) was normalized to 100%. Cold activation profiles of CHO cells expressing human and rat TRPA1 were characterized utilizing cold-induced Cauptake assay. Cauptake by CHO cells and CHO cells transfected with human TRPA1 in response to stimulation with temperatures between 3.5 and 25°C. Cold temperature (4°C) activation of un-induced and tetracycline-induced CHO cells transfected with either human or rat TRPA1. Concentration-dependent inhibition of cold (4°C) activation by ruthenium red in CHO cells expressing human and rat TRPA1. Each point in the graph is an average ± SEM of an experiment conducted in duplicate

Trichloro(sulfanyl)ethyl benzamides inhibit AITC (80 μM) induced inward currents in CHO cells stably expressing human TRPA1

<p><b>Copyright information:</b></p><p>Taken from "Species-specific pharmacology of Trichloro(sulfanyl)ethyl benzamides as transient receptor potential ankyrin 1 (TRPA1) antagonists"</p><p>http://www.molecularpain.com/content/3/1/39</p><p>Molecular Pain 2007;3():39-39.</p><p>Published online 17 Dec 2007</p><p>PMCID:PMC2222611.</p><p></p> Representative traces of inward currents evoked by AITC in the absence or presence of AMG9090 , AMG5445 , AMG2504 , and AMG7160 are shown. ICvalue for each compound was determined from their concentration-dependent inhibition of AITC-induced currents using a PatchXpress 7000A workstation

Fused Piperidines as a Novel Class of Potent and Orally Available Transient Receptor Potential Melastatin Type 8 (TRPM8) Antagonists

The transient receptor potential melastatin type 8 (TRPM8) is a nonselective cation channel primarily expressed in a subpopulation of sensory neurons that can be activated by a wide range of stimuli, including menthol, icilin, and cold temperatures (<25 °C). Antagonism of TRPM8 is currently under investigation as a new approach for the treatment of pain. As a result of our screening efforts, we identified tetrahydrothienopyridine <b>4</b> as an inhibitor of icilin-induced calcium influx in CHO cells expressing recombinant rat TRPM8. Exploration of the structure–activity relationships of <b>4</b> led to the identification of a potent and orally bioavailable TRPM8 antagonist, tetrahydroisoquinoline <b>87</b>. Compound <b>87</b> demonstrated target coverage in vivo after oral administration in a rat pharmacodynamic model measuring the prevention of icilin-induced wet-dog shakes (WDS)

Small Molecule Disruptors of the Glucokinase–Glucokinase Regulatory Protein Interaction: 5. A Novel Aryl Sulfone Series, Optimization Through Conformational Analysis

The glucokinase–glucokinase regulatory protein (GK-GKRP) complex plays an important role in controlling glucose homeostasis in the liver. We have recently disclosed a series of arylpiperazines as in vitro and in vivo disruptors of the GK-GKRP complex with efficacy in rodent models of type 2 diabetes mellitus (T2DM). Herein, we describe a new class of aryl sulfones as disruptors of the GK-GKRP complex, where the central piperazine scaffold has been replaced by an aromatic group. Conformational analysis and exploration of the structure–activity relationships of this new class of compounds led to the identification of potent GK-GKRP disruptors. Further optimization of this novel series delivered thiazole sulfone <b>93</b>, which was able to disrupt the GK-GKRP interaction in vitro and in vivo and, by doing so, increases cytoplasmic levels of unbound GK

Small Molecule Disruptors of the Glucokinase–Glucokinase Regulatory Protein Interaction: 3. Structure–Activity Relationships within the Aryl Carbinol Region of the <i>N</i>‑Arylsulfonamido‑<i>N</i>′‑arylpiperazine Series

We have recently reported a novel approach to increase cytosolic glucokinase (GK) levels through the binding of a small molecule to its endogenous inhibitor, glucokinase regulatory protein (GKRP). These initial investigations culminated in the identification of 2-(4-((2<i>S</i>)-4-((6-amino-3-pyridinyl)­sulfonyl)-2-(1-propyn-1-yl)-1-piperazinyl)­phenyl)-1,1,1,3,3,3-hexafluoro-2-propanol (<b>1</b>, AMG-3969), a compound that effectively enhanced GK translocation and reduced blood glucose levels in diabetic animals. Herein we report the results of our expanded SAR investigations that focused on modifications to the aryl carbinol group of this series. Guided by the X-ray cocrystal structure of compound <b>1</b> bound to hGKRP, we identified several potent GK–GKRP disruptors bearing a diverse set of functionalities in the aryl carbinol region. Among them, sulfoximine and pyridinyl derivatives <b>24</b> and <b>29</b> possessed excellent potency as well as favorable PK properties. When dosed orally in <i>db</i>/<i>db</i> mice, both compounds significantly lowered fed blood glucose levels (up to 58%)

Optimization of Potency and Pharmacokinetic Properties of Tetrahydroisoquinoline Transient Receptor Potential Melastatin 8 (TRPM8) Antagonists

Transient receptor potential melastatin 8 (TRPM8) is a nonselective cation channel expressed in a subpopulation of sensory neurons in the peripheral nervous system. TRPM8 is the predominant mammalian cold temperature thermosensor and is activated by cold temperatures ranging from 8 to 25 °C and cooling compounds such as menthol or icilin. TRPM8 antagonists are being pursued as potential therapeutics for treatment of pain and bladder disorders. This manuscript outlines new developments in the SAR of a lead series of 1,2,3,4-tetrahydroisoquinoline derivatives with emphasis on strategies to improve pharmacokinetic properties and potency. Selected compounds were profiled in two TRPM8 target-specific in vivo coverage models in rats (the icilin-induced wet dog shake model and the cold pressor test). Compound <b>45</b> demonstrated robust efficacy in both pharmacodynamic models with ED₉₀ values <3 mg/kg

Small Molecule Disruptors of the Glucokinase–Glucokinase Regulatory Protein Interaction: 2. Leveraging Structure-Based Drug Design to Identify Analogues with Improved Pharmacokinetic Profiles

In the previous report, we described the discovery and optimization of novel small molecule disruptors of the GK-GKRP interaction culminating in the identification of <b>1</b> (AMG-1694). Although this analogue possessed excellent in vitro potency and was a useful tool compound in initial proof-of-concept experiments, high metabolic turnover limited its advancement. Guided by a combination of metabolite identification and structure-based design, we have successfully discovered a potent and metabolically stable GK-GKRP disruptor (<b>27</b>, AMG-3969). When administered to <i>db</i>/<i>db</i> mice, this compound demonstrated a robust pharmacodynamic response (GK translocation) as well as statistically significant dose-dependent reductions in fed blood glucose levels