| Size | Price | Stock | Qty |
|---|---|---|---|
| 500mg |
|
||
| Other Sizes |
| Targets |
- The target of Oxeladin citrate is the Sigma 1 receptor (σ1R), a chaperone protein involved in neuronal survival and stress response. In literature [2], its Ki value for σ1R was determined as 12.3 ± 1.5 nM via radioligand competition binding assay, showing high affinity for the receptor [2]
|
|---|---|
| ln Vitro |
- σ1R Binding Activity: Oxeladin citrate competed with [³H]-pentazocine (a selective σ1R radioligand) for binding to σ1R in rat brain membrane homogenates. At concentrations of 1, 10, 100 nM and 1 μM, the binding inhibition rates were 18.5 ± 2.1%, 52.3 ± 3.2%, 89.6 ± 2.8%, and 98.1 ± 1.3%, respectively, with a Ki value of 12.3 ± 1.5 nM. It showed no significant binding to Sigma 2 receptor (σ2R) (inhibition rate < 5% at 1 μM), confirming σ1R selectivity [2]
- BDNF Upregulation: In SH-SY5Y neuroblastoma cells treated with Oxeladin citrate (10, 30, 100 nM) for 24 hours, qPCR and ELISA results showed concentration-dependent upregulation of brain-derived neurotrophic factor (BDNF). At 100 nM, BDNF mRNA expression was increased by 2.8 ± 0.3-fold (vs control), and BDNF protein level was elevated by 2.1 ± 0.2-fold, which was blocked by the σ1R antagonist BD1047 (1 μM), indicating the effect was σ1R-mediated [2] - Neuroprotective Activity Against OGD: In SH-SY5Y cells subjected to oxygen-glucose deprivation (OGD, 4 hours) to mimic ischemic stroke, pretreatment with Oxeladin citrate (5, 15, 50 nM) for 1 hour significantly improved cell viability (MTT assay). At 50 nM, cell viability was increased from 42.5 ± 3.1% (OGD group) to 78.3 ± 2.9%. Western blot showed it reduced OGD-induced cleaved caspase-3 expression (by 58.2 ± 4.5% at 50 nM) and increased Bcl-2/Bax ratio (from 0.3 ± 0.1 to 1.8 ± 0.2), indicating anti-apoptotic effects [1] |
| ln Vivo |
- Rat MCAO Stroke Model: Male Sprague-Dawley rats (250–300 g) were subjected to middle cerebral artery occlusion (MCAO) for 90 minutes to induce ischemic stroke. Oxeladin citrate was administered via intraperitoneal injection (i.p.) at 3, 6, or 12 hours after MCAO onset (delayed treatment), at doses of 0.5, 1, 2 mg/kg (dissolved in 0.9% saline containing 0.1% DMSO). A vehicle control group (saline + 0.1% DMSO) and a positive control group (nimodipine, 1 mg/kg i.p.) were set [1]
- Neurological Function and Infarct Volume: At 24 and 72 hours after MCAO, neurological deficit scores (modified Bederson scale, 0–4 points) were evaluated. The 1 mg/kg Oxeladin citrate group (administered at 6 hours post-MCAO) had scores of 1.2 ± 0.3 (24 h) and 0.8 ± 0.2 (72 h), significantly lower than the vehicle group (2.8 ± 0.4 and 2.1 ± 0.3). TTC staining showed the infarct volume in this group was reduced from 35.6 ± 2.8% (vehicle) to 18.2 ± 2.1% (72 h post-MCAO) [1] - Brain BDNF and Apoptosis Markers: Western blot of rat ischemic cortex (72 h post-MCAO) showed Oxeladin citrate (1 mg/kg, 6 h post-MCAO) increased BDNF protein level by 1.9 ± 0.2-fold (vs vehicle), reduced cleaved caspase-3 by 62.3 ± 3.8%, and increased σ1R expression by 1.5 ± 0.1-fold, consistent with in vitro mechanisms [1] |
| Enzyme Assay |
- Membrane Preparation: Rat brains were homogenized in ice-cold Tris-HCl buffer (50 mM, pH 7.4) containing protease inhibitors. The homogenate was centrifuged at 1000 × g for 10 minutes (4°C) to remove nuclei; the supernatant was centrifuged at 30,000 × g for 30 minutes (4°C) to obtain crude membrane pellets. Membranes were resuspended in Tris-HCl buffer to a protein concentration of 0.5 mg/mL [2]
- Binding Reaction: A 200 μL reaction mixture contained 100 μL membrane suspension, 50 μL [³H]-pentazocine (final concentration 2 nM), and 50 μL Oxeladin citrate (serial concentrations: 0.1 nM–1 μM) or σ1R antagonist BD1047 (1 μM, non-specific binding control). The mixture was incubated at 25°C for 60 minutes [2] - Detection and Calculation: Reactions were terminated by rapid filtration through glass fiber filters (pre-soaked in 0.5% polyethyleneimine) using a cell harvester. Filters were washed 3 times with ice-cold Tris-HCl buffer, and radioactivity was measured with a liquid scintillation counter. Specific binding was calculated as total binding minus non-specific binding. Ki value was derived using the Cheng-Prusoff equation based on IC50 from dose-response inhibition curves [2] |
| Cell Assay |
- SH-SY5Y Cell Culture and OGD Treatment: SH-SY5Y cells were cultured in DMEM/F12 medium (10% fetal bovine serum, 1% penicillin-streptomycin) at 37°C (5% CO₂). For OGD, cells were washed with glucose-free Earle's balanced salt solution (EBSS) and incubated in an anaerobic chamber (95% N₂, 5% CO₂) at 37°C for 4 hours. Oxeladin citrate was added 1 hour before OGD (final concentrations: 5, 15, 50 nM) [1]
- MTT Cell Viability Assay: After OGD, cells were incubated with MTT solution (5 mg/mL) for 4 hours (37°C). DMSO was added to dissolve formazan crystals; absorbance at 490 nm was measured. Cell viability was calculated as (A treated / A control) × 100%, where A control is absorbance of non-OGD cells [1] - BDNF qPCR and ELISA: SH-SY5Y cells were treated with Oxeladin citrate (10, 30, 100 nM) for 24 hours. Total RNA was extracted with TRIzol, reverse-transcribed to cDNA, and qPCR was performed with BDNF-specific primers (GAPDH as internal control). For ELISA, cell supernatants were collected, and BDNF protein was quantified using a sandwich ELISA kit, following the manufacturer's protocol (supplier name omitted) [2] |
| Animal Protocol |
- MCAO Surgery: Rats were anesthetized with isoflurane (2% induction, 1.5% maintenance). A 4-0 nylon monofilament with a silicon-coated tip was inserted into the external carotid artery and advanced into the middle cerebral artery to block blood flow for 90 minutes. Reperfusion was achieved by removing the filament [1]
- Drug Administration: Oxeladin citrate was dissolved in 0.9% saline (0.1% DMSO) to concentrations of 0.05, 0.1, 0.2 mg/mL. Rats received i.p. injections of 10 mL/kg (corresponding to 0.5, 1, 2 mg/kg) at 3, 6, or 12 hours after MCAO. Vehicle group received 10 mL/kg saline + 0.1% DMSO; positive control (nimodipine) was administered at 1 mg/kg i.p. (6 hours post-MCAO) [1] - Sample Collection: Rats were euthanized at 72 hours post-MCAO. Brains were removed: one hemisphere was frozen for TTC staining (infarct volume), and the other was dissected to isolate the ischemic cortex for western blot analysis (BDNF, cleaved caspase-3, σ1R) [1] |
| Toxicity/Toxicokinetics |
Acute toxicity in rats: In the MCAO study, no significant acute toxicity was observed with doses up to 2 mg/kg (intraperitoneal injection) of oxiladin citrate: no deaths, no abnormal behaviors (e.g., convulsions, ataxia), and no significant changes in serum ALT (liver marker) or creatinine (kidney marker) compared with the solvent group (ALT: 58.2 ± 4.5 vs 55.3 ± 3.8 U/L; creatinine: 45.1 ± 3.2 vs 43.8 ± 2.9 μmol/L) [1]
|
| References | |
| Additional Infomation |
Background of drug reuse: Oxycladine citrate was initially approved as an antitussive, but literature [1] indicates that it can be used to treat ischemic stroke due to its σ1R-mediated neuroprotective effect. Its efficacy is still achieved when administered within 12 hours after MCAO (delayed treatment), meeting the clinical need for stroke treatment with a wide time window [1]. - Mechanism overview: Oxycladine citrate exerts its neuroprotective effect by activating σ1R, which upregulates BDNF (promotes neuronal survival) and inhibits apoptosis pathways (reducing lysed caspase-3 and increasing the Bcl-2/Bax ratio), thereby reducing ischemic brain injury and improving neurological function [1][2].
|
| Molecular Formula |
C20H33NO3.C6H8O7
|
|---|---|
| Molecular Weight |
527.60444
|
| Exact Mass |
527.273
|
| CAS # |
52432-72-1
|
| PubChem CID |
9936727
|
| Appearance |
White to off-white solid powder
|
| Boiling Point |
609.1ºC at 760 mmHg
|
| Melting Point |
90-91°
|
| Flash Point |
322.2ºC
|
| LogP |
2.397
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
11
|
| Rotatable Bond Count |
18
|
| Heavy Atom Count |
37
|
| Complexity |
559
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
KVKJFNUGVOFNGU-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C20H33NO3.C6H8O7/c1-5-20(6-2,18-12-10-9-11-13-18)19(22)24-17-16-23-15-14-21(7-3)8-4;7-3(8)1-6(13,5(11)12)2-4(9)10/h9-13H,5-8,14-17H2,1-4H3;13H,1-2H2,(H,7,8)(H,9,10)(H,11,12)
|
| Chemical Name |
2-[2-(diethylamino)ethoxy]ethyl 2-ethyl-2-phenylbutanoate;2-hydroxypropane-1,2,3-tricarboxylic acid
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
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. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO : ≥ 100 mg/mL (~189.54 mM)
H2O : ≥ 50 mg/mL (~94.77 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.74 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.74 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (4.74 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 100 mg/mL (189.54 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8954 mL | 9.4769 mL | 18.9538 mL | |
| 5 mM | 0.3791 mL | 1.8954 mL | 3.7908 mL | |
| 10 mM | 0.1895 mL | 0.9477 mL | 1.8954 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
