 Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C20H23CL2F2N3O |
| Molecular Weight | 430.31892991066 |
| Exact Mass | 429.118 |
| CAS # | 2436760-81-3 |
| Related CAS # | Befiradol;208110-64-9 |
| PubChem CID | 135397148 |
| Appearance | Typically exists as white to off-white solids at room temperature |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 28 |
| Complexity | 502 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | MEFWJLIGVNWMAB-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C20H22ClF2N3O.ClH/c1-14-2-4-16(25-11-14)12-24-13-20(23)6-8-26(9-7-20)19(27)15-3-5-18(22)17(21)10-15;/h2-5,10-11,24H,6-9,12-13H2,1H3;1H |
| Chemical Name | (3-chloro-4-fluorophenyl)-[4-fluoro-4-[[(5-methylpyridin-2-yl)methylamino]methyl]piperidin-1-yl]methanone;hydrochloride |
| Synonyms | Befiradol hydrochloride; Befiradol (hydrochloride); Befiradol hydrochloride (208110-64-9 free base); NLX-112 hydrochloride; 2436760-81-3; F 13640 hydrochloride; F 13640 (hydrochloride) |
| 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 | 5-HT1A Receptor |
| ln Vitro | F13640 (befiradol) is a novel 5-HT(1A) receptor agonist with exceptional selectivity vs. other receptors and binding sites[1]. F13640 activates both 5-HT(1A) autoreceptors and postsynaptic 5-HT(1A) receptors in prefrontal cortex with a similar potency. Both activities are likely involved in the analgesic properties of the compound. |
| ln Vivo | Befiradol (F13640; NLX-112) raises the discharge rate of 80% of mPFC pyramidal neurons in the same dose range (ED50=0.62 μg/kg, iv) and decreases the activity of dorsal raphe serotonergic neurons at 0.2-18.2 μg/kg, iv (cumulative doses; ED50=0.69 μg/kg, iv). Subsequent injection of the 5-HT1A receptor antagonist (±)WAY100635 reverses both effects. Befiradol (F13640; NLX-112) (0.04 -0.63 mg/kg, ip) dose-dependently reduces extracellular 5-HT in the mPFC and hippocampal regions in microdialysis experiments. Similarly, Befiradol (F13640; NLX-112) (0.01-2.5 mg/kg, ip) raises extracellular DA in mPFC in a dose-dependent manner. a result reliant on the mPFC's postsynaptic 5-HT1A receptors being activated. In a concentration-dependent manner, local perfusion of Befiradol in mPFC (1-1,000 μM) likewise raises extracellular DA. Befiradol's local and systemic effects can be avoided by administering (±)WAY100635 beforehand[1]. |
| Animal Protocol | Rats were anaesthetized with chloral hydrate (400–500 mg kg−1, i.p.) or isoflurane. A guide cannula with a dummy probe was stereotaxically implanted into the mPFC, stereotaxic coordinates: AP +3.0 mm, L +0.8 mm, DV −1.7 mm, or the hippocampus: AP −4.8 mm, L +4.6 mm, DV −4.6 mm, from bregma and skull surface. Following surgery and recovery from anesthesia, animals were returned to their home cages. At the end of the day, each rat was placed in a microdialysis cage. On the following day, the dummy probe was replaced by a microdialysis probe (3 mm length, 0.5 mm diameter; CMA, Microdialysis AB). The probe was continuously perfused (1.1 μl min−1) with artificial CSF (aCSF) containing 1 μM citalopram for the measure of 5-HT. At least 2 h after probe insertion, samples were collected every 20 min with the first four samples used for baseline. For the experiment with systemic administration of the compounds, saline or (±)WAY100635 were injected s.c., followed, 40 min later, by i.p. administration of saline or F13640. For the experiments with local perfusion, saline was injected s.c. and 40 min later, F13640 was added to the perfusion medium for the concentration–response experiment. For the antagonism, (±)WAY100635 (or aCSF) was delivered through the dialysis probe and 40 min later, F13640 was added to the perfusion medium. Samples were collected for 140 min after administration or beginning of the perfusion of the agonist. At the end of the experiment, rats were killed by anesthetic overdose (pentobarbital 160 mg kg−1, i.p.) and the brain was removed, frozen and cut in a cryomicrotome (Jung Frigocut 2800) to verify the placement of the probe.[1] |
| ADME/Pharmacokinetics | NLX-112 (i.e., F13640, befiradol) exhibits nanomolar affinity, exceptional selectivity and full agonist efficacy at serotonin 5-HT1A receptors. NLX-112 shows efficacy in rat, marmoset and macaque models of L-DOPA induced dyskinesia (LID) in Parkinson's disease and has shown clinical efficacy in a Phase 2a proof-of-concept study for this indication. Here we investigated, in rats, its pharmacodynamic, pharmacokinetic (PK) and brain 5-HT1A receptor occupancy profiles, and its PK properties in the absence and presence of L-DOPA. Total and free NLX-112 exposure in plasma, CSF and striatal ECF was dose-proportional over the range tested (0.04, 0.16 and 0.63 mg/kg i.p.). NLX-112 exposure increased rapidly (Tmax 0.25-0.5h) and exhibited approximately threefold longer half-life in brain than in plasma (1.1 and 3.6h, respectively). At a pharmacologically relevant dose of 0.16 mg/kg i.p., previously shown to elicit anti-LID activity in parkinsonian rats, brain concentration of NLX-112 was 51-63 ng/g from 0.15 to 1h. In microPET imaging experiments, NLX-112 showed dose-dependent reduction of 18F-F13640 (i.e., 18F-NLX-112) brain 5-HT1A receptor labeling in cingulate cortex and striatum, regions associated with motor control and mood, with almost complete inhibition of labeling at the dose of 0.63 mg/kg i.p.. Co-administration of L-DOPA (6 mg/kg s.c., a dose used to elicit LID in parkinsonian rats) together with NLX-112 (0.16 mg/kg i.p.) did not modify PK parameters in rat plasma and brain of either NLX-112 or L-DOPA. Here, we demonstrate that NLX-112's profile is compatible with 'druggable' parameters for CNS indications, and the results provide measures of brain concentrations and 5-HT1A receptor binding parameters relevant to the anti-dyskinetic activity of the compound.https://pubmed.ncbi.nlm.nih.gov/39096379/ |
| References |
[1]. In vivo electrophysiological and neurochemical effects of the selective 5-HT1A receptor agonist, F13640, at pre- and postsynaptic 5-HT1A receptors in the rat. Psychopharmacology (Berl). 2012 May;221(2):261-72. |
| Additional Infomation |
Rationale: F13640 (befiradol) is a novel 5-HT(1A) receptor agonist with exceptional selectivity vs. other receptors and binding sites. It shows analgesic activity in animal models and is currently developed for human use.[1] Objectives: Given the potential dual role of the serotonergic system in pain, through the modulation of ascending signals in spinal cord and their emotional processing by corticolimbic areas, we examined the in vivo activity of F13640 at somatodendritic autoreceptors and postsynaptic 5-HT(1A) heteroreceptors in medial prefrontal cortex (mPFC).[1] Methods: In vivo single unit recordings and intracerebral microdialysis in the rat.[1] Results: F13640 reduced the activity of dorsal raphe serotonergic neurons at 0.2-18.2 μg kg(-1), i.v. (cumulative doses; ED(50) = 0.69 μg kg(-1), i.v.) and increased the discharge rate of 80% of mPFC pyramidal neurons in the same dose range (ED(50) = 0.62 μg kg(-1), i.v.). Both effects were reversed by the subsequent administration of the 5-HT(1A) receptor antagonist (±)WAY100635. In microdialysis studies, F13640 (0.04-0.63 mg kg(-1), i.p.) dose-dependently decreased extracellular 5-HT in the hippocampus and mPFC. Likewise, F13640 (0.01-2.5 mg kg(-1), i.p.) dose-dependently increased extracellular DA in mPFC, an effect dependent on the activation of postsynaptic 5-HT(1A) receptors in mPFC. Local perfusion of F13640 in mPFC (1-1,000 μM) also increased extracellular DA in a concentration-dependent manner. Both the systemic and local effects of F13640 were prevented by prior (±)WAY100635 administration.[1] Conclusions: These results indicate that, upon systemic administration, F13640 activates both 5-HT(1A) autoreceptors and postsynaptic 5-HT(1A) receptors in prefrontal cortex with a similar potency. Both activities are likely involved in the analgesic properties of the compound. |
Solubility Data
| Solubility (In Vitro) | DMSO: 125 mg/mL (290.48 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.83 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 (4.83 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 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (4.83 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3239 mL | 11.6193 mL | 23.2385 mL | |
| 5 mM | 0.4648 mL | 2.3239 mL | 4.6477 mL | |
| 10 mM | 0.2324 mL | 1.1619 mL | 2.3239 mL |