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    ANA-12
    ANA-12

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    This product is for research use only, not for human use. We do not sell to patients.
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    InvivoChem Cat #: V0642
    CAS #: 219766-25-3Purity ≥98%

    Description: ANA-12 is a novel, potent and selective TrkB antagonist with Kd of 10 nM and 12 μM for the high and low affinity sites, respectively. 

    References: J Clin Invest. 2011 May 2; 121(5): 1846–1857; Eur J Neurosci. 2014 May;39(9):1439-54.

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    Molecular Weight (MW)407.49
    FormulaC22H21N3O3S
    CAS No.219766-25-3
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 37 mg/mL (90.79 mM)
    Water: <1 mg/mL
    Ethanol:  <1 mg/mL
    Solubility (In vivo)

    Chemical Name: N-(2-((2-oxoazepan-3-yl)carbamoyl)phenyl)benzo[b]thiophene-2-carboxamide

    InChi Key: TUSCYCAIGRVBMD-UHFFFAOYSA-N

    InChi Code: InChI=1S/C22H21N3O3S/c26-20(25-17-10-5-6-12-23-21(17)27)15-8-2-3-9-16(15)24-22(28)19-13-14-7-1-4-11-18(14)29-19/h1-4,7-9,11,13,17H,5-6,10,12H2,(H,23,27)(H,24,28)(H,25,26)

    SMILES Code: O=C(C1=CC2=CC=CC=C2S1)NC3=CC=CC=C3C(NC4C(NCCCC4)=O)=O

    Synonyms

    ANA 12; ANA-12; ANA12; 


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    In Vitro

    In vitro activity: ANA-12 binds directly and selectively to TrkB and inhibits processes downstream of TrkB without altering TrkA and TrkC functions. In nnr5 PC12-TrkB cells, ANA-12 prevents brain-derived neurotrophic factor (BDNF)-induced neurite outgrowth at concentrations as low as 10 nM. In DRG neurons, ANA-12 eliminates the effect of BDNF on increasing inward current.


    Kinase Assay: Maxisorp ELISA 96-well plates are coated with various concentrations of Trk BECD -Fc, 20 mg/ml BSA, or 1 mg/mL IgG-Fc (polyclonal anti-TrkB) in a carbonate buffer (pH 9.6) overnight at 4°C. Plates are saturated with 0.5% BSA in PBS for 2 hours at room temperature and extensively washed in PBS-Tween 0.05%. Bodipy–ANA-12 is then incubated in 0.5% PBS-BSA for 1 hour at room temperature before the addition of BDNF in 0.5% PBS-BSA for another hour. After extensive washes in PBS-Tween 0.05%, the amount of bodipy–ANA-12 bound is quantified by fluorescence at 520 ± 10 nm. Detectability range for extrapolation analysis is assessed by coating ELISA plates with bodipy–ANA-12 and reading fluorescence at 520 ± 10 nm.


    Cell Assay:  Modulation of neurite outgrowth by molecules is assessed in nnr5 PC12–TrkB, –TrkA, and –TrkC cells after addition of BDNF (1 nM), NGF (2 nM), and NT-3 (10 nM), respectively. The number of cells bearing neurites longer than 2 cells in diameter in each counting field is microscopically determined (2 fields per well, 3 wells per condition). Counting is performed blind each 24 hours for 3 days.

    In VivoIn adult C57BL6/129SveV F1s mice, ANA-12 (0.5 mg/kg, i.p.) decreases TrKB activity in the brain and reduces the anxiety- and depression-related behaviors without affecting the neuron survival. In male C57BL/6 mice, ANA-12 (0.5 mg/kg, i.p.) shows antidepressant-like effects on lipopolysaccharide-induced depression-like behavior. In male Sprague-Dawley rats, ANA-12 (3 μg/dose) blocks the effect of medial nucleus tractus solitarius (mNTS) BDNF on reducing food intake. In male wild-type mice, ANA-12 reverses ethanol intake and induces D3 receptor downregulation, but is ineffective in D3R-/- mice. In male CocSired rats, ANA-12 (0.5 mg/kg, i.p.) reverses the diminished self-administration of cocaine.
    Animal modelAdult C57BL6/129SveV F1s mice
    Formulation & DosageDissolved in 1% DMSO dissolved in 0.9% NaCl solution; 0.5 mg/kg; i.p.
    References

    J Clin Invest. 2011 May 2; 121(5): 1846–1857; Eur J Neurosci. 2014 May;39(9):1439-54; Int J Neuropsychopharmacol. 2014 Oct 31;18(4). pii: pyu077.


    These protocols are for reference only. InvivoChem does not independently validate these methods.

    ANA-12

    Effects of 7,8-DHF and ANA-12 on LPS-induced changes in phosphorylation of TrkB in the mouse brain. Int J Neuropsychopharmacol. 2015 Feb; 18(4): pyu077.

    ANA-12

    Role of TrkB and mTORC1 in the antidepressant action of 7,8-DHF and ANA-12 on LPS-induced depression-like behavior. Int J Neuropsychopharmacol. 2014 Oct 31;18(4).

    ANA-12

    Role of TrkB and mTORC1 in the antidepressant action of 7,8-DHF and ANA-12 on LPS-induced depression-like behavior. Int J Neuropsychopharmacol. 2014 Oct 31;18(4).


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