Abstract
Transient receptor potential melastatin type 8 (TRPM8) is a target for the treatment of different physio-pathological processes. While TRPM8 antagonists are reported as potential drugs for pain, cancer, and inflammation, to date only a limited number of chemotypes have been investigated and thus a limited number of compounds have reached clinical trials. Hence there is high value in searching for new TRPM8 antagonistic to broaden clues to structure-activity relationships, improve pharmacological properties and explore underlying molecular mechanisms. To address this, the EDASA Scientific in-house molecular library has been screened in silico, leading to identifying twenty-one potentially antagonist compounds of TRPM8. Calcium fluorometric assays were used to validate the in-silico hypothesis and assess compound selectivity. Four compounds were identified as selective TRPM8 antagonists, of which two were dual-acting TRPM8/TRPV1 modulators. The most potent TRPM8 antagonists (BB 0322703 and BB 0322720) underwent molecular modelling studies to highlight key structural features responsible for drug–protein interaction. The two compounds were also investigated by patch-clamp assays, confirming low micromolar potencies. The most potent compound (BB 0322703, IC50 1.25 ± 0.26 μM) was then profiled in vivo in a cold allodinya model, showing pharmacological efficacy at 30 μM dose. The new chemotypes identified showed remarkable pharmacological properties paving the way to further investigations for drug discovery and pharmacological purposes.
Highlights
Transient receptor potential melastatin type 8 (TRPM8) is emerging as a potential target for the treatment of a variety of disorders, given its key role in many physio-pathological processes [1]
Compounds were initially flexibly docked using Glide SP, and the best scoring ones were advanced to the following step that refined and rescored the docking poses using Glide XP, giving a list of compounds ranked according to their XP GlideScore values, that ranged from −11.982 to −8.903
Two of these compounds showed low micromolar potencies when challenged in patch-clamp measurements, with compound 14 (BB 0322703) exhibiting in vivo efficacy in a model of oxaliplatin-induced cold allodynia
Summary
Transient receptor potential melastatin type 8 (TRPM8) is emerging as a potential target for the treatment of a variety of disorders, given its key role in many physio-pathological processes [1] It is a non-selective Ca2+ permeable channel that shows a highly complicated gating behaviour regulated by multiple factors [2,3] including temperature, endogenous and exogenous modulators or mechanisms, and depolarizing voltages [4,5]. Structure-based virtual screening represents an outstanding alternative tool for putative hit compounds discovery, allowing the investigation of large chemical libraries through computational methods by applying knowledge about the protein target [32] This approach has been profitably used for the identification of several chemotypes as TRPM8 antagonists [25,33]. The effect of one of these compounds was studied in an in vivo model of cold allodynia, revealing its efficacy and suitability for further development
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