Identification of a low–molecular weight TrkB antagonist with anxiolytic and antidepressant activity in mice
Maxime Cazorla, … , Christoph Kellendonk, Didier Rognan
Maxime Cazorla, … , Christoph Kellendonk, Didier Rognan
Published April 18, 2011
Citation Information: J Clin Invest. 2011;121(5):1846-1857. https://doi.org/10.1172/JCI43992.
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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 5

ANA-12 directly binds and selectively modulates TrkB.

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ANA-12 directly binds and selectively modulates TrkB. (A) Bodipy–ANA-12 ...
(A) Bodipy–ANA-12 (100 μM) was incubated with increasing amounts of TrkBECD-Fc, IgG-Fc, BSA, or nothing (negative control; dashed line). Specific binding could be detected only with TrkBECD-Fc. Data represent mean ± SEM of 2 experiments performed in triplicate. (B) Increasing concentrations of bodipy–ANA-12 were incubated with TrkBECD-Fc (1 μg/ml) in the presence (filled circles) or absence (filled squares) of 10 nM BDNF. Values are normalized to maximal signal after subtraction of nonspecific binding (BSA or IgG-Fc). Affinity constants (Kd) were derived from Scatchard plotting of the data (inset, right panel). Data represent mean ± SEM of 4 experiments performed in triplicate. (C) Computational model of the docking of ANA-12 (cyan) into the specificity patch of TrkB-d5 (green ribbon). 3 putative hydrogen bonds (red dotted lines) anchor ANA-12 to TrkB. Note that ANA-12 is surrounded by TrkB-specific amino acid residues (red) in the binding pocket. (D and E) Representative photomicrographs and quantitative analyses of neurotrophin-induced neurite outgrowth in the presence of ANA-12 in nnr5 PC12–TrkB (D), –TrkA, and –TrkC (E) cells. Original magnification, ×20. Data presented for TrkA and TrkC are those obtained with 100 μM of ANA-12 and are not different from their respective controls. #P < 0.01 and ‡P < 0.0001. Data represent mean ± SEM of 3 experiments performed in triplicate.