CNQX Disodium (FG-9065), the disodium salt form of CNQX, is a potent AMPA/kainate antagonist with higher water-solubility than the free acid form CNQX (Cat. No. V33762). As an AMPA and kainate receptor antagonist, it inhibits AMPA and kainate receptors with IC50 values of 0.3 μM, 1.5 μM, respectively. CNQX is also an antagonist at the glycine modulatory site on the NMDA receptor complex with an IC50 of 25 μM). CNQX can be used to isolate GABAA receptor mediated spontaneous inhibitory postsynaptic currents and antagonizes non-NMDA receptor-mediated responses in cultured cerebellar granule cells. CNQX shows neuroprotective effects in models of ischemia and inhibits seizure-like activity in hippocampal neurons.
Physicochemical Properties
| Molecular Formula | C9H2N4NA2O4 |
| Molecular Weight | 276.11 |
| Exact Mass | 275.987 |
| Elemental Analysis | C, 39.15; H, 0.73; N, 20.29; Na, 16.65; O, 23.18 |
| CAS # | 479347-85-8 |
| Related CAS # | CNQX;115066-14-3 |
| PubChem CID | 6093155 |
| Appearance | Brown to red solid powder |
| Hydrogen Bond Donor Count | 0 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 0 |
| Heavy Atom Count | 19 |
| Complexity | 362 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | YCXDDPGRZKUGDG-UHFFFAOYSA-L |
| InChi Code | InChI=1S/C9H4N4O4.2Na/c10-3-4-1-5-6(2-7(4)13(16)17)12-9(15)8(14)11-5;;/h1-2H,(H,11,14)(H,12,15);;/q;2*+1/p-2 |
| Chemical Name | disodium;6-cyano-7-nitroquinoxaline-2,3-diolate |
| Synonyms | FG9065; FG-9065; CNQX (disodium); disodium;6-cyano-7-nitroquinoxaline-2,3-diolate; 6-CYANO-7-NITROQUINOXALINE-2,3-DIONE DISODIUM; 7-Nitro-2,3-dioxo-1,2,3,4-tetrahydroquinoxaline-6-carbonitrile, disodium salt; CNQX disodium salt?; CNQX Disodium; CNQX 2Na; FG 9065 |
| 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) |
Biological Activity
| Targets | AMPA (IC50 = 0.3 μM); kainate receptor (IC50 = 1.5 μM)[1] |
| ln Vitro | In hippocampal slice perfusion, CNQX disodium (FG9065 disodium; 2–5 μM) reversibly suppresses mossy fiber excitatory postsynaptic potentials (EPSPs) and Schaffer collaterals while avoiding both slow and fast GABA-mediated inhibition[2]. A specific and dose-dependent reduction in the amplitude of the monosynaptic component of DR-VRR recorded from lumbar segments is observed with CNQX disodium (1–5 μM) [3]. |
| ln Vivo |
In the first 15 minutes of the cocaine-free phase, CNQX disodium (FG9065 disodium; 0.75–3 mg/kg; i.p.; 20 minutes before to testing) dose-dependently decreases the number of cocaine reactions [4]. Ten minutes prior to the retention test, CNQX disodium (0.5 or 1.25 μg) can be infused bilaterally into the amygdala or dorsal hippocampus to partially decrease the expression of hypotensive inhibitory avoidance in rats 24 hours post-training. At 0.5 μg, CNQX disodium completely blocks the channel [5]. The motor pattern underlying locomotion in the lamprey is activated and maintained by excitatory amino acid neurotransmission. The quinoxalinediones 6,7-dinitroquinoxaline-2,3-dione (DNQX) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) are potent and selective antagonists of non-N-methyl-D-aspartate (NMDA) receptors in the mammalian central nervous system. In the lamprey, these compounds are now shown to block fast excitatory synaptic potentials elicited in neurones of the spinal ventral horn. They selectively antagonise responses to the application of selective kainate and quisqualate receptor agonists (kainate and alpha-amino-3-hydroxy-5-methyl-4-isoxalone (AMPA)) but do not influence NMDA receptor-mediated responses. Additionally, it is shown that the activation of NMDA receptors is sufficient to elicit and maintain fictive locomotion after blockade of non-NMDA receptors with either DNQX or CNQX. Conversely, activation of quisqualate receptors with AMPA, but not quisqualate leads to fictive locomotion with properties much like that activated by kainate[3]. Increasing the cocaine unit dose increased responding during the first and second intervals, with a decrease in the latency to the first CS. CNQX decreased the number of cocaine responses in a dose-dependent manner during the first 15-min cocaine-free interval, but did not affect cocaine responding during either the second interval or the latter part of the session under the FR4(FR7:S) schedule. In the locomotor activity test, reductions in rearing were produced by higher CNQX doses than those that attenuated significantly responding during the first fixed interval. Conclusions: These results suggest that AMPA/kainate receptors are involved in mediation of cocaine-seeking behaviour controlled partly by cocaine-associated cues[4]. The involvement of non-N-methyl-D-aspartate receptors in the amygdala in the expression of conditioned fear was examined using the fear-potentiated startle paradigm. Rats implanted with bilateral cannulae in the basolateral amygdaloid nuclei received 10 pairings of either a visual or auditory conditioned stimulus with footshock on each of 2 days. The next day, they were tested by eliciting the acoustic startle reflex in the presence or absence of the conditioned stimulus and divided into groups with equivalent levels of potentiation. One or two days later, rats were tested again following intra-amygdala infusion of vehicle or 0.025, 0.25, or 2.5 micrograms of 6-cyano-7-nitroquinoxaline-2,3-dione. The drug dose-dependently blocked the expression of potentiated startle in both sensory modalities, indicating that activation of non-NMDA receptors in the amygdala is necessary for the expression of conditioned fear[5]. |
| Enzyme Assay | Superfusion of hippocampal slices with 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 2-5 microM) reversibly blocked the Schaffer collateral and mossy fibre excitatory postsynaptic potential (EPSP), while sparing the fast and slow gamma-aminobutyric acid (GABA)-mediated inhibition. Membrane potential, input resistance and spike accommodation were not altered. Inward currents induced by quisqualate were reduced to a greater extent by CNQX than those induced by kainate or N-methyl-D-aspartate. We suggest that CNQX may be a useful antagonist to study excitatory amino acid-mediated synaptic transmission[2]. |
| Animal Protocol |
Animal/Disease Models: Male Wistar rats, body weight 180-200 g[4] Doses: 0.75, 1.5 and 3 mg/kg Route of Administration: IP; 20 minutes before test Experimental Results: During the first 15-minute cocaine-free interval, cocaine (IV; 0.25 mg/infusion) The number of reactions is diminished in a dose-dependent manner. Free-feeding Wistar rats were trained to respond for an IV cocaine infusion (0.25 mg/infusion) under a FI15 min(FR7:S) schedule, whereby the completion of FR7 responses led to the presentation of a conditioned stimulus (CS). After two 15-min fixed intervals, rats were allowed to respond for cocaine under an FR4(FR7:S) second-order schedule for another 120 min. After acquisition of stable responding, the cocaine unit dose was increased to 0.50 mg/infusion. The effects of CNQX (0, 0.75, 1.5, and 3 mg/kg IP) on cocaine seeking were then examined using a within-subjects design.[4] |
| References |
[1]. Quinoxalinediones: Potent Competitive non-NMDA Glutamate Receptor Antagonists. Science. 1988 Aug 5;241(4866):701-3. [2]. Blockade of excitatory synaptic transmission by 6-cyano-7-nitroquinoxaline-2,3-dione(CNQX) in the hippocampus in vitro. Neurosci Lett. 1988 Sep 23;92(1):64-8. [3]. CNQX and DNQX block non-NMDA synaptic transmission but not NMDA-evoked locomotion in lamprey spinal cord. Brain Res. 1990 Jan 8;506(2):297-302. [4]. Attenuation of Cocaine-Seeking Behaviour by the AMPA/kainate Receptor Antagonist CNQX in Rats. Psychopharmacology (Berl). 2003 Feb;166(1):69-76. [5]. Infusion of the non-NMDA receptor antagonist CNQX into the amygdala blocks the expression of fear-potentiated startle. Behav Neural Biol. 1993 Jan;59(1):5-8. |
| Additional Infomation | The N-methyl-D-aspartate (NMDA)-subtype of glutamate receptors has been well described as a result of the early appearance of NMDA antagonists, but no potent antagonist for the "non-NMDA" glutamate receptors has been available. Quinoxalinediones have now been found to be potent and competitive antagonists at non-NMDA glutamate receptors. These compounds will be useful in the determination of the structure-activity relations of quisqualate and kainate receptors and the role of such receptors in synaptic transmission in the mammalian brain.[1] |
Solubility Data
| Solubility (In Vitro) | DMSO : ~10.53 mg/mL (~38.14 mM) |
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.6217 mL | 18.1087 mL | 36.2174 mL | |
| 5 mM | 0.7243 mL | 3.6217 mL | 7.2435 mL | |
| 10 mM | 0.3622 mL | 1.8109 mL | 3.6217 mL |