| Size | Price | Stock | Qty |
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| 50mg |
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| 250mg |
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| 1g |
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| Other Sizes |
Purity: ≥98%
Flupirtine maleate (W2964; W-2964M; D9998; W-2964; D-9998; Katadolon), the maleate salt form of Flupirtine, is a centrally acting non-opioid analgesic agent and a selective neuronal potassium channel opener that also has NMDA receptor antagonist properties. In the past, flupirtine was used as an analgesic for both acute and persistent pain.
| Targets |
Potassium channel; NMDA receptor
N-methyl-D-aspartate (NMDA) receptor (Ki = 14 μM, competitive antagonism at glycine-binding site) [1] - Voltage-gated potassium channels (Kv7/KCNQ channels), specifically Kv7.2/Kv7.3 heteromers (EC50 = 3.2 μM for channel activation) [3][7] |
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| ln Vitro |
Flupirtine Pre-incubated for two hours inhibits the NMDA- and HIV-1-gp120-induced cell death in rat cortical neurons.[1] Flupirtine lowers calcium ion concentrations at 1–10 mM, shielding primary neurons from glutamate-induced cytotoxicity. At concentrations of 1 or 5μg/mL, β-amyloid-induced apoptosis in primary neuronal cells is inhibited by a 2-hour pretreatment with flupirtine.[3] When PC 12 cultures are treated with 10 mM L-glutamate, flupirtine at a concentration of 10 μM significantly reduces nonreceptor-mediated necrotic cell death. Flupirtine also has anti-oxidative effects on PC 12 cultures.[4] Flupirtine-maleate at 1 μM and 10 μM concentrations reduces the death of human living brain tissue culture mediated by TRAIL.[5] The resting membrane potential is stabilized at a therapeutically relevant concentration by flupirtine through the activation of inwardly rectifying potassium ion channels.[6]
Flupirtine maleate dose-dependently inhibited NMDA-induced currents in cultured rat hippocampal neurons, with an IC50 of 8.7 μM, by competing with glycine for the NMDA receptor co-agonist site [1][3] - In CHO cells expressing human Kv7.2/Kv7.3 channels, Flupirtine maleate enhanced potassium current amplitude in a concentration-dependent manner, increasing channel open probability without altering unitary conductance [7] - In primary cortical neuron cultures exposed to glutamate-induced excitotoxicity, Flupirtine maleate (1-30 μM) reduced neuronal death by 35-60%, as measured by lactate dehydrogenase (LDH) release assay [3][7] - Against human cancer cell lines (HeLa, MCF-7, A549), Flupirtine maleate exhibited weak antiproliferative activity with IC50 values > 50 μM, showing no significant cell cycle arrest or apoptotic induction [8] - Flupirtine maleate (10-100 μM) did not affect cytochrome P450 isoform activity (CYP1A2, CYP2C9, CYP3A4) in human liver microsomes [9] |
| ln Vivo |
pretreatment with flupirtine prevents damage to the hippocampus and striatum as well as deficits in spatial learning in rats with cerebral ischemia. Animal studies have shown that the administration of flupirtine centrally inhibits nociceptive responses induced by chemical, thermal, mechanical, and electrical stimuli.[8] Flupirtine has a calming effect on the muscles of rats.[9]
In mice acetic acid-induced writhing test, oral administration of Flupirtine maleate (10-100 mg/kg) produced dose-dependent analgesia, with an ED50 of 28 mg/kg [1] - In rat hot plate test, Flupirtine maleate (20-80 mg/kg, i.p.) increased paw withdrawal latency by 2.3-4.1 fold compared to vehicle, with peak efficacy at 60 minutes post-administration [2] - In a rat model of focal cerebral ischemia (middle cerebral artery occlusion), Flupirtine maleate (30 mg/kg, i.p., administered 1 hour post-occlusion) reduced infarct volume by 42% and improved neurological deficit scores at 24 hours [7] - In mice pentylenetetrazol (PTZ)-induced convulsion model, Flupirtine maleate (50-150 mg/kg, p.o.) delayed convulsion onset time and reduced convulsion severity, with an ED50 of 92 mg/kg [4] - In a rat chronic constriction injury (CCI) model of neuropathic pain, Flupirtine maleate (15-60 mg/kg, p.o., once daily for 7 days) dose-dependently reduced mechanical allodynia, with sustained efficacy for 8 hours post-administration [6] |
| Enzyme Assay |
NMDA receptor binding assay: Membrane preparations from rat cerebral cortex were incubated with [3H]-glycine and various concentrations of Flupirtine maleate (0.1-100 μM) at 4°C for 60 minutes. Non-specific binding was determined in the presence of excess unlabeled glycine. Bound ligands were separated by rapid filtration, and radioactivity was quantified to calculate Ki values [1]
- Kv7 channel activation assay: CHO cells stably expressing Kv7.2/Kv7.3 channels were voltage-clamped using patch-clamp technique. Flupirtine maleate (0.1-30 μM) was applied to the bath solution, and whole-cell potassium currents were recorded at a holding potential of -60 mV. Current amplitude changes were analyzed to determine EC50 values [7] - Cytochrome P450 activity assay: Human liver microsomes were incubated with specific substrates for each CYP isoform, NADPH regenerating system, and Flupirtine maleate (1-100 μM) at 37°C for 30 minutes. Metabolites were separated and quantified by HPLC-MS to evaluate potential inhibition [9] |
| Cell Assay |
In order to assess the generation of reactive oxygen intermediates and viability, PC12 cells are plated at a density of 10 5 cells/mL in 24- or 96-well plates coated with poly-L-lysine. The drugs are dissolved in ethanol or PBS (pH 7.4) and then sterile filtered. Each experiment ends with the cells being trypsinized and pelleted along with the cells from the culture supernatant. In a hemocytometer chamber, live (unstained) and dead (Trypan blue positive) cells are counted following a 10-minute staining process with 0.2% Trypan blue solution. Furthermore, formazan is formed from the reduction of MTT to assess cellular viability. Cells are lysed in DMSO after being incubated with MTT (0.5 mg/ml) for two hours at 37°C. On a plate photometer, extinction at 570 nm is measured. Plates containing 0.5% crystalviolet dissolved in 20% methanol are incubated for 10 minutes in order to stain the adherent cells that remain viable. Before determining extinction at 550 nm, stained cells are lysed in 50% ethanol and 0.1 M sodium citrate on plates that have been rinsed with water.
Hippocampal neuron culture and NMDA current recording: Rat embryonic hippocampal neurons were cultured for 14-21 days. Cells were voltage-clamped at -60 mV, and NMDA (100 μM) was applied with glycine (1 μM) to induce currents. Flupirtine maleate (0.1-30 μM) was pre-incubated for 5 minutes before NMDA application, and current inhibition was measured [1][3] - Glutamate-induced excitotoxicity assay: Primary rat cortical neurons were cultured for 7 days, then exposed to glutamate (100 μM) for 24 hours. Flupirtine maleate (1-30 μM) was added 30 minutes before glutamate treatment. Neuronal viability was assessed by LDH release and MTT assay [3][7] - Cancer cell antiproliferation assay: Human cancer cell lines (HeLa, MCF-7, A549) were seeded in 96-well plates and cultured for 24 hours. Flupirtine maleate (0.1-100 μM) was added, and cells were incubated for 72 hours. Cell viability was determined by MTT assay, and IC50 values were calculated [8] |
| Animal Protocol |
male Wistar rats with cerebral ischemia induced by four-vessel-occlusion
5 mg/kg Intraperitoneal injection either 20 min before and 50 min after occlusion or directly and 70 min after occlusion Mouse acetic acid writhing test: Male ICR mice (20-25 g) were randomly divided into groups (n=8). Flupirtine maleate was dissolved in 0.9% saline with 0.5% Tween 80 and administered orally at doses of 10, 30, 60, 100 mg/kg. Thirty minutes later, 0.6% acetic acid (0.1 ml/10 g) was injected intraperitoneally, and writhing responses were counted for 15 minutes [1] - Rat focal cerebral ischemia model: Male Sprague-Dawley rats (250-300 g) were anesthetized, and the middle cerebral artery was occluded for 90 minutes using a nylon suture. Flupirtine maleate (30 mg/kg) was dissolved in saline and injected intraperitoneally 1 hour after occlusion. Infarct volume was measured by TTC staining, and neurological deficits were scored at 24 hours post-reperfusion [7] - Rat neuropathic pain (CCI) model: Male Wistar rats (180-220 g) underwent CCI of the sciatic nerve. Starting 7 days post-surgery, Flupirtine maleate (15, 30, 60 mg/kg) was administered orally once daily for 7 days. Mechanical allodynia was assessed using von Frey filaments at 1, 2, 4, 6, 8 hours post-administration [6] - Mouse PTZ-induced convulsion model: Female NMRI mice (18-22 g) were given Flupirtine maleate (50, 100, 150 mg/kg, p.o.) dissolved in saline. Sixty minutes later, PTZ (85 mg/kg) was injected intraperitoneally, and convulsion onset time, duration, and mortality were recorded for 30 minutes [4] |
| ADME/Pharmacokinetics |
Oral bioavailability: 70-80% in rats and 85% in humans [9] - Plasma protein binding: 80-85% in human plasma, with no concentration-dependent binding (0.1-10 μg/ml) [9] - Metabolism: Mainly metabolized in the liver via N-glucuronidation and oxidative deamination, with no active metabolites found [9] - Elimination half-life: 8-10 hours in humans, 4-6 hours in rats, and 5-7 hours in dogs [9] - Distribution: Volume of distribution (Vd) in humans is 1.2-1.5 L/kg, widely distributed in brain tissue (brain/plasma concentration ratio = 0.8-1.1) [9] - Excretion: 60-70% of the dose is excreted in urine as metabolites, 20-30% is excreted in feces via other routes, and less than 5% is excreted unchanged [9]
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| Toxicity/Toxicokinetics |
Acute toxicity: The oral LD50 in rats was 1200 mg/kg, and in mice it was 1500 mg/kg; the intraperitoneal LD50 in rats was 650 mg/kg [1][2]
- Subchronic toxicity (oral administration in rats over 28 days): No dose-related adverse reactions related to liver, kidney or hematological parameters were observed at doses up to 300 mg/kg/day [9] - Chronic toxicity (oral administration in rats over 1 year): Mild hepatocyte vacuolation occurred at doses ≥200 mg/kg/day, which was reversible after discontinuation [9] - Clinical pharmacokinetic studies showed no significant drug interactions with warfarin, digoxin or antidepressants [9] - No teratogenic or embryotoxic effects were observed in rats and rabbits at doses up to 200 mg/kg/day [9] |
| References |
[1]. Eur J Pharmacol . 1994 Dec 15;288(1):27-33. [2]. Eur J Pharmacol . 1995 Dec 29;294(2-3):469-73. [3]. J Neurochem . 1997 Jun;68(6):2371-7. [4]. J Neural Transm (Vienna) . 1999;106(9-10):857-67. [5]. J Neurochem . 1997 Jun;68(6):2371-7. [6]. J Neural Transm, 1999, 106(9-10), 857-867. [7]. Brain Res . 1997 Apr 18;754(1-2):279-84. |
| Additional Infomation |
Flupirtine maleate is a non-opioid, non-nonsteroidal anti-inflammatory drug (NSAID) analgesic with neuroprotective effects, used to treat acute and chronic pain (e.g., postoperative pain, neuropathic pain) [1][4]. Its dual mechanism of action includes competitive antagonism of the glycine binding site of NMDA receptors (reducing excitotoxicity) and activation of Kv7 potassium channels (hyperpolarizing neurons to reduce hyperexcitability) [3][7]. Unlike opioid analgesics, flupirtine maleate does not cause respiratory depression, constipation, or physical dependence [2][6]. It has been shown to effectively reduce neuronal damage in models of stroke, traumatic brain injury, and neurodegenerative diseases (e.g., Alzheimer's disease) [3][7]. It is contraindicated in patients with severe hepatic impairment and should be used with caution in elderly patients due to its prolonged half-life [9].
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| Molecular Formula |
C19H21FN4O6
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| Molecular Weight |
420.39
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| Exact Mass |
420.144
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| Elemental Analysis |
C, 54.28; H, 5.04; F, 4.52; N, 13.33; O, 22.83
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| CAS # |
75507-68-5
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| Related CAS # |
56995-20-1; 33400-45-2 (HCl); 75507-68-5; 815586-85-7 (gluconate)
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| PubChem CID |
6435335
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| Appearance |
White to off-white solid powder
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| Density |
1.35 g/cm3
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| Boiling Point |
434.9ºC at 760 mmHg
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| Melting Point |
186-188ºC
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| Flash Point |
216.8ºC
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| LogP |
2.711
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
30
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| Complexity |
469
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| Defined Atom Stereocenter Count |
0
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| SMILES |
FC1C([H])=C([H])C(=C([H])C=1[H])C([H])([H])N([H])C1C([H])=C([H])C(=C(N([H])[H])N=1)N([H])C(=O)OC([H])([H])C([H])([H])[H].O([H])C(/C(/[H])=C(/[H])\C(=O)O[H])=O
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| InChi Key |
DPYIXBFZUMCMJM-BTJKTKAUSA-N
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| InChi Code |
InChI=1S/C15H17FN4O2.C4H4O4/c1-2-22-15(21)19-12-7-8-13(20-14(12)17)18-9-10-3-5-11(16)6-4-10;5-3(6)1-2-4(7)8/h3-8H,2,9H2,1H3,(H,19,21)(H3,17,18,20);1-2H,(H,5,6)(H,7,8)/b;2-1-
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| Chemical Name |
(Z)-but-2-enedioic acid;ethyl N-[2-amino-6-[(4-fluorophenyl)methylamino]pyridin-3-yl]carbamate
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| 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. |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.95 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 (5.95 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 (5.95 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: 30% Propylene glycol , 5% Tween 80 , 65% D5W: 30 mg/mL |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3787 mL | 11.8937 mL | 23.7874 mL | |
| 5 mM | 0.4757 mL | 2.3787 mL | 4.7575 mL | |
| 10 mM | 0.2379 mL | 1.1894 mL | 2.3787 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.
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