Identification of a low–molecular weight TrkB antagonist with anxiolytic and antidepressant activity in mice
Maxime Cazorla, Joël Prémont, Andre Mann, Nicolas Girard, Christoph Kellendonk, Didier Rognan
Maxime Cazorla, Joël Prémont, Andre Mann, Nicolas Girard, Christoph Kellendonk, Didier Rognan
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Research Article Neuroscience

Identification of a low–molecular weight TrkB antagonist with anxiolytic and antidepressant activity in mice

Abstract

The neurotrophin brain-derived neurotrophic factor (BDNF) and its receptor tropomyosin-related kinase B (TrkB) have emerged as key mediators in the pathophysiology of several mood disorders, including anxiety and depression. However, therapeutic compounds that interact with TrkB receptors have been difficult to develop. Using a combination of structure-based in silico screening and high-capacity functional assays in recombinant and neuronal cells, we identified a low–molecular weight TrkB ligand (ANA-12) that prevented activation of the receptor by BDNF with a high potency. ANA-12 showed direct and selective binding to TrkB and inhibited processes downstream of TrkB without altering TrkA and TrkC functions. KIRA-ELISA analysis demonstrated that systemic administration of ANA-12 to adult mice decreased TrkB activity in the brain without affecting neuronal survival. Mice administered ANA-12 demonstrated reduced anxiety- and depression-related behaviors on a variety of tests predictive of anxiolytic and antidepressant properties in humans. This study demonstrates that structure-based virtual screening strategy can be an efficient method for discovering potent TrkB-selective ligands that are active in vivo. We further propose that ANA-12 may be a valuable tool for studying BDNF/TrkB signaling and may constitute a lead compound for developing the next generation of therapeutic agents for the treatment of mood disorders.

Authors

Maxime Cazorla, Joël Prémont, Andre Mann, Nicolas Girard, Christoph Kellendonk, Didier Rognan

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Figure 2

Pharmacological properties of N-T04 and N-T19 in different systems.

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Pharmacological properties of N-T04 and N-T19 in different systems. (A) ...
(A) Structures of the 2 most active compounds, N-T04 and N-T19. (B and C) KIRA-ELISA assessment of N-T04 and N-T19 in absence (filled squares) or presence (filled circles) of BDNF in TetOn-rhTrkB and neuronal cells. Values were normalized to signal obtained with 1 nM (B) or 0.4 nM (C) of BDNF after subtraction of signal obtained in control wells. Compounds were compared with 10 μM of K252a (open squares), a concentration that abolishes basal TrkB activity (20). IC50 values of both high-affinity (4.7 ± 1.9 μM) and low-affinity (231 ± 53 μM) sites for N-T19 in neurons were determined using Eadie-Hofstee plotting of the data (lower panels). Nonlinear regression curve fit for N-T19 in neurons was obtained using a 2-site competition model. Data represent mean ± SEM of 3–6 experiments performed in triplicate. (D) BDNF concentration-response curve in absence (squares) or in presence (diamonds, 3 μM; triangles, 30 μM; circles, 300 μM) of N-T04 or N-T19. A noncompetitive antagonism is clearly demonstrated for N-T19 by Eadie-Hofstee plotting of the data (lower panel). Values were normalized to the maximal BDNF response after subtraction of signal obtained in controls. Data represent mean ± SEM of 4 experiments performed in triplicate.