5-HT1A Receptor

F13640（贝维拉多）是一种新型5-HT（1A）受体激动剂，与其他受体和结合位点相比具有特殊的选择性[1]。F13640以相似的效力激活前额叶皮层中的5-HT（1A）自身受体和突触后5-HT（1A）受体。这两种活性都可能与化合物的镇痛特性有关。[1]

Befiradol (F13640; NLX-112) 在相同剂量范围内 (ED50=0.62 μg/kg, iv) 提高 80% mPFC 锥体神经元的放电率，并在 0.2-18.2 μg/kg 时降低中缝背侧血清素能神经元的活性，iv（累积剂量；ED50=0.69 μg/kg，iv）。随后注射 5-HT1A 受体拮抗剂 (±)WAY100635 可逆转这两种效应。在微透析实验中，Befiradol (F13640; NLX-112) (0.04 -0.63 mg/kg, ip) 剂量依赖性地减少 mPFC 和海马区域的细胞外 5-HT。类似地，Befiradol (F13640；NLX-112) (0.01-2.5 mg/kg，腹腔注射) 以剂量依赖性方式提高 mPFC 中的细胞外 DA。该结果依赖于 mPFC 的突触后 5-HT1A 受体被激活。以浓度依赖性方式，在 mPFC 中局部灌注 Befiradol (1-1,000 μM) 同样会提高细胞外 DA。通过预先给予(±)WAY100635可以避免贝非拉多的局部和全身效应[1]。

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]

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/

[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.

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.

C20H23CL2F2N3O

430.31892991066

429.118

2436760-81-3

Befiradol;208110-64-9

135397148

Typically exists as white to off-white solids at room temperature

45.2Ų

2

5

5

28

502

0

MEFWJLIGVNWMAB-UHFFFAOYSA-N

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

(3-chloro-4-fluorophenyl)-[4-fluoro-4-[[(5-methylpyridin-2-yl)methylamino]methyl]piperidin-1-yl]methanone;hydrochloride

Befiradol hydrochloride; Befiradol (hydrochloride); Befiradol hydrochloride (208110-64-9 free base); NLX-112 hydrochloride; 2436760-81-3; F 13640 hydrochloride; F 13640 (hydrochloride)

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 125 mg/mL (290.48 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (4.83 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 20.8 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.08 mg/mL (4.83 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.08 mg/mL (4.83 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。