Amlexanox (AA673; CHX3673)   中文名称: 氨来占诺

IKKε (IC50 = 1-2 μM); TBK1 (IC50 = 1-2 μM)

Amlexanox 具有剂量反应关系，通过磷酸化 MBP 抑制 IKK- 和 TBK1 活性，IC50 约为 1-2 μM。在大致相同的浓度下，amlexanox 也会抑制 TBK1 的活性，但它对 IKK-α 或 IKK-β 没有影响，并且在这些浓度下，它不会抑制代表大多数激酶家族的大组中的任何其他激酶。 Amlexanox 与 IKK-ɛ 或 TBK1 竞争其底物 ATP，表明它与 ATP 结合位点的酶相互作用。当受到聚肌苷酸：聚胞苷酸刺激时，Amlexanox 会增加 3T3-L1 脂肪细胞中 TBK1 Ser172 的磷酸化，并抑制干扰素反应因子 3 (IRF3)（IKK-ɛ 和 TBK1[1] 的假定底物）的磷酸化（多聚 I：C）。

Amlexanox 疗法可减少脂肪变性，提高胰岛素敏感性，并通过增加生热作用使肥胖小鼠体重减轻[1]。

体外激酶测定是通过将纯化的激酶（IKK 或 TBK1）在 0.5 Ci µCi γ-[32P]--ATP 和每个样品 1 g MBP 作为激酶底物的情况下在 30°C 下孵育 30 分钟来进行含有 25 mM Tris (pH 7.5)、10 mM MgCl2、1 mM DTT 和 10 µM ATP 的缓冲液。添加 4x 十二烷基硫酸钠 (SDS) 样品缓冲液后，在 95°C 下煮沸 5 分钟即可停止激酶反应。使用 SDS 聚丙烯酰胺凝胶电泳解析上清液并转移至硝酸纤维素后，使用 Typhoon 9410 磷光成像仪进行放射自显影分析。

使用 Cell Counting Kit-8 查看细胞增殖时，请遵循制造商的说明。 BMM 以每孔 5×103 个细胞的密度接种在 96 孔板中。 24 小时后，每 2 天对细胞进行不同剂量的 AmLexanox（0、1.5、3、6、12、25 μM）处理，同时暴露于 M-CSF (30 ng/mL)，总共 7 天。 1、3、5和7天后将培养基更换为含有10% CCK-8的培养基，然后将细胞在37°C下再孵育2小时。然后在 ELX800 吸光度酶标仪上，在 450 nm 波长下测量吸光度。

In contrast to ND C57BL/6 controls, wildtype male C57BL/6 mice are fed a HFD starting at eight weeks of age for 12–24 weeks that contains 45% of their calories from fat. ω-3 fatty acids are included in the diets fed to C57BL/6 mice. Mice that have been on the HFD for 16 weeks receive a three-week course of rosiglitazone treatment by ingesting the medication. The average daily dose of rosiglitazone consumed by each mouse is 3.5 mg/kg. Every day oral gavage is used to administer AmLexanox. AmLexanox (25 mg or 100 mg per kg) administration for the prevention groups starts at eight weeks of age, concurrent with HFD feeding. After 12 weeks of HFD, the treatment groups start receiving 25 mg per kg of amLexanox at 20 weeks old. After eight weeks of amLexanox treatment, mice in the treatment group are switched from receiving amLexanox gavage to receiving a vehicle control to test the effects of amLexanox withdrawal. At ten weeks of age, control and ob/ob mice receive a standard chow diet along with 100 mg per kg of amLexanox or a vehicle control. The animals are kept in a facility that is specifically pathogen-free, has a 12-hour light/12-hour dark cycle, and they have unrestricted access to food and water.

[1].Nat Med . 2013 Mar;19(3):313-21.

C16H14N2O4

298.2934

298.10

C, 64.42; H, 4.73; N, 9.39; O, 21.45

68302-57-8

68302-57-8

Solid powder

CC(C)C1=CC2=C(C=C1)OC3=NC(=C(C=C3C2=O)C(=O)O)N

SGRYPYWGNKJSDL-UHFFFAOYSA-N

InChI=1S/C16H14N2O4/c1-7(2)8-3-4-12-9(5-8)13(19)10-6-11(16(20)21)14(17)18-15(10)22-12/h3-7H,1-2H3,(H2,17,18)(H,20,21)

2-amino-5-oxo-7-propan-2-ylchromeno[2,3-b]pyridine-3-carboxylic acid

CHX 3673 CHX-3673 CHX3673AA-673 AA673 AA 673 Amoxanox, Aphthasol.

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)

DMSO: 59~100 mg/mL (197.8~335.2 mM)
Ethanol: ~2 mg/mL(~6.7 mM)

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。