AZD-9272

mGluR5

在人和大鼠 mGluR5 表达细胞系中，AZD 9272 会导致细胞内 Ca2+ 对 1.5 μM mGluR I 类选择性激动剂 DHPG 的反应强度出现浓度依赖性下降。最大抑制率为100%。 AZD9272 的人 mGluR5 的平均 IC50 (±SD) 值为 7.6±1.1 nM (n=13)。 AZD9272 的大鼠 mGluR5 的平均 IC50 值为 2.6±0.3 nM (n=3)。相比之下，10 μM AZD9272 不会减弱背景 GHEK 细胞对 10 μM ATP 的反应。 AZD9272 浓度的增加会导致 DHPG 的效力和最大反应降低。 AZD9272 以浓度依赖性方式完全逆转人 mGluR5-GHEK 细胞中谷氨酸刺激的 (EC80, 80 μM) 磷脂酰肌醇水解，IC50 为 26±3 nM (n=21)[1]。

单次静脉注射 3 μmol/kg 剂量后，AZD 9272 的清除率较低，AZD 9272 从血浆中消除，终末半衰期为 2 至 6 小时。口服给药后的终末半衰期与静脉给药后的半衰期相似。 AZD9272 稳态时的分布体积处于中等水平[1]。 AZD9272不会引起可卡因适当的反应，并且在较高剂量下会导致反应率非剂量依赖性降低。 2.84 mg/kg 的 AZD9272 在给药后 20 小时内引起超过 80%、通常超过 99% 的 MTEP 适当反应，在给药后 24 小时下降至约 20%，产生 21.93 小时的 t1/2，并且不会对响应率造成系统性影响。 AZD9272 引起 >90% MTEP 适当反应的第一个时间点是在给药后 30 分钟[2]。

涉及饱和结合和竞争结合的研究利用 22°C 孵育一小时。为了进行饱和实验，将源自 mGluR5-GHEK 细胞的膜置于不同浓度（0.1 至 30 nM）的 [³H]AZD9272（含或不含 10 μM MPEP）中。在这些研究的变体中发现了饱和 [³H]AZD9272 结合，其中 MPEP 以低浓度（10 和 20 nM）存在。无论 MPEP 是否存在，B_max 保持恒定，表明这些配体与单个结合位点相互作用[1]。

在含有 10 µCi/mL [³H]肌醇的培养基中，将 hmGluR5-GHEK 细胞以 50,000 个细胞/孔接种到 96 孔板上。将细胞培养 16 小时，然后漂洗 3 次，并在补充有 2 mM 丙酮酸和 1 单位/mL 谷氨酸丙酮酸转氨酶的 HEPES 缓冲盐水中于 37°C 放置 1 小时。细胞在溶液中洗涤一次后，在含有 10 mM LiCl 的 HEPES 缓冲盐水中预孵育 10 分钟。为了确定拮抗剂活性，首先将细胞与 AZD9272 预孵育 10 分钟，然后与谷氨酸（EC80，80 µM）在 37°C 下孵育 30 分钟。 AZD9272 的十个浓度（范围从 1 nM 到 30 μM）一式两份进行检查。在冰上添加 0.1 mL (5%) 高氯酸并在 4°C 下孵育至少 30 分钟即可结束反应[1]。

In a colony room with lights on from 6:00 AM to 6:00 PM, about 48 male Wistar rats weighing between 240 and 250 g at the start of the experiments are housed in pairs or up to 8 rats per cage. By limiting food access, the animals are kept at about 80% of their free feeding weight. The animals are separated into distinct groups and trained to distinguish between the presence of PCP (1.6 mg/kg i.p., 30 minutes), MTEP (2 mg/kg i.p., 30 minutes), AZD9272 (1.6 mg/kg p.o., 60 minutes), and cocaine (3.4 mg/kg i.p., 15 minutes)[1].

[1]. AZD9272 and AZD2066: selective and highly central nervous system penetrant mGluR5 antagonists characterized by their discriminative effects. J Pharmacol Exp Ther. 2014 Aug;350(2):212-22.

[2]. Discovery and characterization of AZD9272 and AZD6538-Two novel mGluR5 negative allosteric modulators selected for clinical development. Bioorg Med Chem Lett. 2012 Nov 15;22(22):6974-9.

C14H6N4OF2

284.22044

284.05

C, 59.16; H, 2.13; F, 13.37; N, 19.71; O, 5.63

327056-26-8

Solid powder

C1=CC(=NC=C1F)C2=NOC(=N2)C3=CC(=CC(=C3)C#N)F

RBSPCALDSNXWEP-UHFFFAOYSA-N

InChI=1S/C14H6F2N4O/c15-10-1-2-12(18-7-10)13-19-14(21-20-13)9-3-8(6-17)4-11(16)5-9/h1-5,7H

3-fluoro-5-[3-(5-fluoropyridin-2-yl)-1,2,4-oxadiazol-5-yl]benzonitrile

AZD 9272; AZD-9272; AZD9272

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

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.

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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).

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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)

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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).

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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

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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.

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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。