sigma-1 receptor ( IC50 = 0.86 μM ); muscarinic
Sigma 1 (σ1) receptor (IC₅₀ = 0.86 μM) [2]
Muscarinic receptors (M1-M4) (IC₅₀ = 3.3 – 5.2 μM) [2]
Sodium channel site 2 (IC₅₀ = 5.1 μM) [2]
NMDA receptor (IC₅₀ = 8.0 μM) [2]

The pre-administration of AVex-73 hydrochloride (ANAVEX2-73) leads to a dose-dependent attenuation of the scopolamine induced alternation deficit, significant at 1 and 3 mg/kg. The step-through latency impairments are significantly reduced, dose-dependently, at doses greater than 0.3 mg/kg, by pre-treating with AVex-73 hydrochloride[1]. Significant improvement in recognition memory deficit is observed at 1 mg/kg of AVex-73 hydrochloride treatment, which is administered in a dose-dependent manner. AVex-73 hydrochloride, at doses of 0.1 and 1 mg/kg, significantly attenuates the significant Aβ25-35-induced decrease in Akt phosphorylation one day after injections. The reduction in Ser9 phosphorylation caused by the peptide at 0.3 and 1 mg/kg is lessened seven days after injections by AVex-73 hydrochloride. The administration of AVex-73 hydrochloride treatment inhibits the rise in Aβ1-42 content induced by Aβ25-35 in a dose-dependent manner, exhibiting a noteworthy impact at the maximum dosage examined[2].

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Binding assays show that ANAVEX2-73 binds to muscarinic acetylcholine and sigma1 (σ1) receptors with affinities in the low micromolar range. It acts as a potent muscarinic compound, with Ki values lower than 500 nM for all M1–M4 subtypes, and as a moderate σ1 receptor agonist, but is selective towards σ2 subtypes. [1]

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In a nontransgenic mouse model of Alzheimer's disease (intracerebroventricular injection of Aβ₂₅₋₃₅ oligomers), ANAVEX2-73 (0.1, 0.3, 1 mg/kg, i.p.) dose-dependently prevented the recognition memory deficit in the novel object recognition test, with a significant effect observed at 1 mg/kg [2].
ANAVEX2-73 treatment (0.1, 0.3, 1 mg/kg, i.p.) significantly attenuated the Aβ₂₅₋₃₅-induced decrease in Akt phosphorylation (Ser473) in the mouse hippocampus at day 1 post-injection (significant at 0.1 and 1 mg/kg) and showed a non-significant trend toward attenuation at day 7 [2].
At day 7 post-Aβ₂₅₋₃₅ injection, ANAVEX2-73 (0.3, 1 mg/kg, i.p.) attenuated the decrease in GSK-3β phosphorylation at the inhibitory Ser9 site, though not reaching statistical significance. However, it significantly blocked the Aβ₂₅₋₃₅-induced increase in GSK-3β phosphorylation at the activating Tyr216 site at doses of 0.3, and 1 mg/kg [2].
ANAVEX2-73 (0.1-1 mg/kg, i.p.) dose-dependently and significantly blocked Aβ₂₅₋₃₅-induced Tau hyperphosphorylation at physiological (Ser202, Thr205, detected by AT8 antibody) and pathological (Ser212, Thr214, detected by AT100 antibody) epitopes in the mouse hippocampus at day 7 [2].
ANAVEX2-73 (1 mg/kg, i.p.) significantly prevented the Aβ₂₅₋₃₅-induced increase in endogenous Aβ₁₋₄₂ content in the mouse hippocampus at day 7, as measured by ELISA [2].
ANAVEX2-73 (0.3, 1 mg/kg, i.p.) dose-dependently and significantly prevented the Aβ₂₅₋₃₅-induced increase in C99 fragment levels (a β-secretase cleavage product of APP) in the mouse hippocampus at day 7 [2].

The pre-administration of AVex-73 hydrochloride (ANAVEX2-73) leads to a dose-dependent attenuation of the scopolamine induced alternation deficit, significant at 1 and 3 mg/kg. The pre-treatment with AVex-73 hydrochloride attenuates the impairments of step-through latency, dose dependently and significantly at doses higher than 0.3 mg/kg[1]. The AVex-73 hydrochloride treatment dose-dependently blocks the recognition memory deficit, with a significant effect measured at 1 mg/kg. One day after injections, the significant Aβ25-35-induced decrease in Akt phosphorylation is significantly attenuated by AVex-73 hydrochloride at 0.1 and 1 mg/kg dose. Seven days after injections,AVex-73 hydrochloride attenuates the decrease in Ser9 phosphorylation induced by the peptide at 0.3 and 1 mg/kg. The AVex-73 hydrochloride treatment dose-dependently prevents the Aβ25-35-induced increase in Aβ1-42 content, with a significant effect at the highest dose tested[2].

We use male mice that weigh 32±2 g and are 7–9 weeks old. Substances, such as brahmenesine hydrochloride, are diluted to the appropriate dose and injected at a 100 μL/20 g body weight volume. Animals are used for behavioral testing from day 1 to day 9 following intravenous injections, or they are killed prior to biochemical measures.

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Drug Administration for Efficacy Study:** Male Swiss OF-1 mice (7-9 weeks old) received an intracerebroventricular (i.c.v.) injection of either oligomeric Aβ₂₅₋₃₅ peptide (9 nmol) or scrambled Aβ₂₅₋₃₅ peptide as a control. ANAVEX2-73 (0.1, 0.3, or 1 mg/kg) was dissolved in physiological saline and administered intraperitoneally (i.p.) 20 minutes before the i.c.v. peptide injection. Control groups received saline (i.p.) [2].
* **Novel Object Recognition Test:** The test was performed on days 6-8 post-injection. On day 6, mice were habituated to the open field for 10 min (session 1). On day 7, two identical objects were placed in the open field, and exploratory activity was recorded for 10 min (session 2). On day 8, one familiar object was replaced with a novel object, and exploratory activity was recorded for 10 min (session 3). The preferential exploration index (ratio of contacts with the novel object/total contacts) was calculated. Animals with fewer than 10 total contacts were excluded [2].
* **Biochemical Analysis – Tissue Collection:** Mice were euthanized by decapitation at specific time points (1 or 7 days) after peptide and drug injections. The hippocampus was rapidly dissected, frozen in liquid nitrogen, and stored at -80°C for subsequent Western blotting or ELISA analysis [2].
* **Western Blotting:** Hippocampal tissue was homogenized in lysis buffer with protease and phosphatase inhibitors. Proteins (20-50 μg per lane) were separated by SDS-PAGE and transferred to PVDF membranes. Membranes were blocked and incubated overnight at 4°C with primary antibodies against phosphorylated and total forms of Akt, GSK-3β, Tau (AT8, AT100), and C99, as well as β-tubulin as a loading control. After washes, membranes were incubated with HRP-conjugated secondary antibodies. Immunoreactive bands were visualized using ECL and quantified with ImageJ software. Protein expression levels were normalized to total protein or loading controls [2].
* **Aβ₁₋₄₀/₄₂ ELISA:** Hippocampal tissue was homogenized in Tris-NaCl buffer and sonicated. After centrifugation, supernatants were collected. Mouse Aβ₁₋₄₀ and Aβ₁₋₄₂ concentrations were measured using commercially available ELISA kits according to the manufacturer's instructions. Absorbance was read at 450 nm, and sample concentrations were calculated from standard curves and expressed as pg/mg tissue [2].

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Drug Administration for Efficacy Study: Male Swiss OF-1 mice (7-9 weeks old) received an intracerebroventricular (i.c.v.) injection of either oligomeric Aβ₂₅₋₃₅ peptide (9 nmol) or scrambled Aβ₂₅₋₃₅ peptide as a control. ANAVEX2-73 (0.1, 0.3, or 1 mg/kg) was dissolved in physiological saline and administered intraperitoneally (i.p.) 20 minutes before the i.c.v. peptide injection. Control groups received saline (i.p.) [2].
Novel Object Recognition Test: The test was performed on days 6-8 post-injection. On day 6, mice were habituated to the open field for 10 min (session 1). On day 7, two identical objects were placed in the open field, and exploratory activity was recorded for 10 min (session 2). On day 8, one familiar object was replaced with a novel object, and exploratory activity was recorded for 10 min (session 3). The preferential exploration index (ratio of contacts with the novel object/total contacts) was calculated. Animals with fewer than 10 total contacts were excluded [2].
Biochemical Analysis – Tissue Collection: Mice were euthanized by decapitation at specific time points (1 or 7 days) after peptide and drug injections. The hippocampus was rapidly dissected, frozen in liquid nitrogen, and stored at -80°C for subsequent Western blotting or ELISA analysis [2].
Western Blotting: Hippocampal tissue was homogenized in lysis buffer with protease and phosphatase inhibitors. Proteins (20-50 μg per lane) were separated by SDS-PAGE and transferred to PVDF membranes. Membranes were blocked and incubated overnight at 4°C with primary antibodies against phosphorylated and total forms of Akt, GSK-3β, Tau (AT8, AT100), and C99, as well as β-tubulin as a loading control. After washes, membranes were incubated with HRP-conjugated secondary antibodies. Immunoreactive bands were visualized using ECL and quantified with ImageJ software. Protein expression levels were normalized to total protein or loading controls [2].
Aβ₁₋₄₀/₄₂ ELISA: Hippocampal tissue was homogenized in Tris-NaCl buffer and sonicated. After centrifugation, supernatants were collected. Mouse Aβ₁₋₄₀ and Aβ₁₋₄₂ concentrations were measured using commercially available ELISA kits according to the manufacturer's instructions. Absorbance was read at 450 nm, and sample concentrations were calculated from standard curves and expressed as pg/mg tissue [2].

Preliminary metabolism studies indicate that ANAVEX2-73 is transformed to a single major metabolite, ANAVEX19-144, through demethylation of the tertiary amine group. This metabolite is pharmacologically active, contributing to the drug's long duration of action, as the parent compound is effective when administered up to 12 hours before an amyloid challenge. In contrast, the metabolite of a related compound, ANAVEX1-41 (ANAVEX2-140), is inactive. [1]

[1]. Anti-amnesic and neuroprotective potentials of the mixed muscarinic receptor/sigma 1 (σ1) ligand ANAVEX2-73, a novel aminotetrahydrofuran derivative. J Psychopharmacol. 2011 Aug;25(8):1101-17.

[2]. Blockade of Tau Hyperphosphorylation and Aβ1-42 Generation by the Aminotetrahydrofuran Derivative ANAVEX2-73, a Mixed Muscarinic andσ1 Receptor Agonist, in a Nontransgenic Mouse Model of Alzheimer's Disease. Neuropsychopharmacology. 2013 Aug; 38(9): 1706-1723.

See also: Blarcamesine (notes moved to).

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ANAVEX2-73 (tetrahydro-N,N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride) is an aminotetrahydrofuran derivative characterized as a mixed σ1 receptor and muscarinic receptor agonist with a moderate affinity range (sub-micromolar for σ1, micromolar for muscarinic receptors) [2].
The compound has been shown to be neuroprotective in Alzheimer's disease models in vivo and was in clinical phase I/IIa at the time of this publication [2].
In the Aβ₂₅₋₃₅ mouse model, ANAVEX2-73 demonstrated an ability to block both Tau hyperphosphorylation (via modulation of the Akt/GSK-3β pathway) and the seeding of endogenous Aβ₁₋₄₂, suggesting a potential disease-modifying activity [2].
The effects of ANAVEX2-73 are attributed to the synergistic activation of both muscarinic (particularly M1) and σ1 receptors. Muscarinic activation can stimulate the PLC/PKC pathway, which inhibits GSK-3β, while σ1 activation can modulate Ca²⁺ signaling and potentially also influence kinase activities and secretase function [2].
Despite its moderate (micromolar) affinity for its targets, ANAVEX2-73 showed in vivo efficacy at low mg/kg doses comparable to selective compounds with nanomolar affinities, which may help limit adverse effects often seen with pure, high-affinity muscarinic agonists [2].

C19H24CLNO

317.857

317.154

C, 71.80; H, 7.61; Cl, 11.15; N, 4.41; O, 5.03

195615-84-0

Blarcamesine; 195615-83-9; 195615-84-0 (HCl)

46932299

White to off-white solid powder

1

2

4

22

298

0

Cl[H].O1C([H])([H])C([H])([H])C([H])(C([H])([H])N(C([H])([H])[H])C([H])([H])[H])C1(C1C([H])=C([H])C([H])=C([H])C=1[H])C1C([H])=C([H])C([H])=C([H])C=1[H]

FEQOLYDPQKHFTD-UHFFFAOYSA-N

InChI=1S/C19H23NO.ClH/c1-20(2)15-18-13-14-21-19(18,16-9-5-3-6-10-16)17-11-7-4-8-12-17;/h3-12,18H,13-15H2,1-2H3;1H

1-(2,2-diphenyloxolan-3-yl)-N,N-dimethylmethanamine;hydrochloride

Anavex2-73; Anavex 2-73; Anavex-2-73; Anavex-273; AE-37 hydrochloride; Anavex273; Anavex 273

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 25~64 mg/mL (78.7~201.4 mM)
Water: ~64 mg/mL
Ethanol: ~3 mg/mL

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.87 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (7.87 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (7.87 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 100 mg/mL (314.61 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)