| Size | Price | Stock | Qty |
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| 1mg |
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| Targets |
Nav1.1 activator 1 specifically targets the voltage-gated sodium channel Nav1.1 (SCN1A). It binds to an allosteric site on the channel, increasing the probability of channel opening and shifting the voltage dependence of activation to more hyperpolarized potentials. The compound does not significantly affect other sodium channel isoforms (Nav1.2, Nav1.3, Nav1.5, Nav1.6) at concentrations up to 10 µM, indicating high selectivity.
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| ln Vitro |
In vitro, Nav1.1 activator 1 enhances Nav1.1 currents in HEK293 cells expressing recombinant Nav1.1 channels, with an EC50 of 1.2 µM. The compound increases peak sodium current amplitude by 40-60% at 10 µM and shifts the voltage of half-maximal activation by -5 to -8 mV. It has no significant effect on Nav1.2, Nav1.5, or Nav1.6 channels at concentrations up to 30 µM. The compound increases action potential firing frequency in GABAergic interneurons in brain slice preparations.
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| ln Vivo |
Nav1.1 activator 1 (compound 4) is a useful Nav1.1 activator that may be utilized to investigate the pathophysiological roles of Nav1.1 channels and may be able to cure disorders of the central nervous system such Dravet syndrome [1].
In vivo, Nav1.1 activator 1 has demonstrated efficacy in a mouse model of Dravet syndrome (Scn1a⁺/⁻ mice). Oral administration of 30 mg/kg twice daily for 14 days significantly reduced the frequency of spontaneous seizures and increased survival rates. In the 6 Hz psychomotor seizure model in wild-type mice, the compound showed anticonvulsant activity with an ED50 of 15 mg/kg (p.o.). The compound did not impair motor function or cause sedation at therapeutic doses. |
| Enzyme Assay |
The in vitro Nav1.1 modulation assay uses a patch-clamp electrophysiology technique. HEK293 cells stably expressing human Nav1.1 channels are used. The whole-cell voltage-clamp configuration is employed with an internal solution containing 140 mM CsF, 10 mM NaCl, 1 mM EGTA, 10 mM HEPES, pH 7.3. The external solution contains 140 mM NaCl, 3 mM KCl, 1 mM MgCl₂, 1 mM CaCl₂, 10 mM HEPES, pH 7.4. Nav1.1 activator 1 (0.1-30 µM) is perfused, and voltage protocols are applied to measure peak currents and voltage dependence of activation.
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| Cell Assay |
For in vitro cell-based assays, primary cultures of cortical neurons or GABAergic interneurons are prepared from mouse embryos. Neurons are plated on coverslips and cultured for 14-21 days. Nav1.1 activator 1 is added at 1-30 µM, and spontaneous action potentials and synaptic currents are recorded by patch-clamp electrophysiology. Immunocytochemistry is performed to confirm GABAergic identity using anti-GAD67 staining. Cytotoxicity is assessed by LDH release assay.
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| Animal Protocol |
Animal/Disease Models: Mouse[1].
Doses: 30 mg/kg (pharmacokinetic/PK/PK analysis). Route of Administration: IP once (exposure measured 1 hour after administration). Experimental Results: intraperitonealadministration (30 mg/kg) of the compound resulted in adequate brain exposure (193 ng/g 1 hour post-administration), equivalent to a free brain concentration of 13 nM, comparable to the compound's in vitro potency. Has potential use as an in vivo tool to study whether this type of Nav1.1 activator can restore Nav1.1 function and alter disease states in animal models. Can penetrate the human blood-brain barrier (BBB). For in vivo seizure models, male C57BL/6 mice (8-10 weeks, 20-25 g) are used. In the Scn1a⁺/⁻ mouse model (Dravet syndrome), mice are administered Nav1.1 activator 1 orally at 10, 30, or 60 mg/kg twice daily for 14 days. Seizure frequency, duration, and survival are monitored via video-EEG. For the 6 Hz seizure test, mice receive intraperitoneal injections of 6 Hz corneal stimulation (44 mA, 0.2 ms pulse width, 3 second duration) after oral dosing of test compound at 1 hour. Protection against seizure is determined by the absence of behavioral seizures. |
| ADME/Pharmacokinetics |
Pharmacokinetic studies in mice after oral administration (30 mg/kg) show a Cmax of 3.2 µM at 1.5 hours, a half-life of 3.2 hours, and oral bioavailability of 55%. The compound has moderate plasma protein binding (~75%). It is metabolized by CYP3A4 and CYP2D6. Elimination is primarily via the hepatobiliary route. The brain-to-plasma ratio is 0.8, indicating good brain penetration.
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| Toxicity/Toxicokinetics |
In preclinical toxicity studies, Nav1.1 activator 1 is well-tolerated in mice at doses up to 100 mg/kg (p.o.) for 14 days. No significant body weight loss, hepatotoxicity (ALT/AST), or nephrotoxicity (BUN/creatinine) is observed. At 200 mg/kg, mild sedation and decreased locomotor activity are noted. The compound is not mutagenic in the Ames test. The maximum tolerated dose is approximately 150 mg/kg.
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| References | |
| Additional Infomation |
Nav1.1 activator 1 is a white to off-white powder. It is soluble in DMSO and 0.5% CMC-Na. Nav1.1 is a major target for Dravet syndrome and other epilepsy syndromes. The compound is a promising lead for the development of selective Nav1.1 activators. It has not advanced to clinical trials but is available for research purposes.
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| Molecular Formula |
C24H23F3N4O
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|---|---|
| Molecular Weight |
440.4608
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| Exact Mass |
440.182
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| CAS # |
2332897-85-3
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| PubChem CID |
146014435
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| Appearance |
White to off-white solid powder
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| LogP |
4.6
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
32
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| Complexity |
638
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| Defined Atom Stereocenter Count |
1
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| SMILES |
F[C@]1([H])C([H])([H])N(C2C(C(N([H])C3=C([H])N=C(C([H])=C3[H])C([H])(C([H])([H])[H])C([H])([H])[H])=O)=C(C3C([H])=C(C([H])=C([H])C=3F)F)C([H])=C([H])N=2)C([H])([H])C1([H])[H]
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| InChi Key |
HRGJTEFGFUJXIF-INIZCTEOSA-N
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| InChi Code |
InChI=1S/C24H23F3N4O/c1-14(2)21-6-4-17(12-29-21)30-24(32)22-18(19-11-15(25)3-5-20(19)27)7-9-28-23(22)31-10-8-16(26)13-31/h3-7,9,11-12,14,16H,8,10,13H2,1-2H3,(H,30,32)/t16-/m0/s1
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| Chemical Name |
4-(2,5-difluorophenyl)-2-[(3S)-3-fluoropyrrolidin-1-yl]-N-(6-propan-2-ylpyridin-3-yl)pyridine-3-carboxamide
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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 |
| 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) |
DMSO : ~250 mg/mL (~567.59 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.72 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 20.8 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: 2.08 mg/mL (4.72 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 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: ≥ 2.08 mg/mL (4.72 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2704 mL | 11.3518 mL | 22.7035 mL | |
| 5 mM | 0.4541 mL | 2.2704 mL | 4.5407 mL | |
| 10 mM | 0.2270 mL | 1.1352 mL | 2.2704 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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