| 规格 | 价格 | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| 靶点 |
Human mAChR M4: Atropine acts as an antagonist with an IC50 of 390 pM and a calculated Ki(CP) of 140 pM in a CRE-luciferase assay using transfected HEK293T cells. [3]
Chicken mAChR cM4: Atropine acts as an antagonist with an IC50 of 710 pM and a calculated Ki(CP) of 120 pM in a CRE-luciferase assay using transfected HEK293T cells. [3] Human alpha2A-adrenoceptor (hADRA2A): Atropine acts as an antagonist with an IC50 of 45 μM and a calculated Ki(CP) of 14 μM in a CRE-luciferase assay using transfected HEK293T cells. [3] |
|---|---|
| 体外研究 (In Vitro) |
在一项使用转染了人源M4 mAChR的CRISPR-M3 HEK293T细胞进行的CRE-荧光素酶实验中，阿托品抑制了卡巴胆碱(10 μM)诱导的荧光信号，IC50值为390 pM。由此数据计算出的抑制常数(Ki)为140 pM。[3]
在一项使用转染了鸡源M4 mAChR (cM4)的CRISPR-M3 HEK293T细胞进行的CRE-荧光素酶实验中，阿托品抑制了卡巴胆碱(10 μM)诱导的荧光信号，IC50值为710 pM。由此数据计算出的抑制常数(Ki)为120 pM。[3] 在一项使用转染了人源alpha2A-肾上腺素能受体 (hADRA2A)的CRISPR-M3 HEK293T细胞进行的CRE-荧光素酶实验中，阿托品抑制了可乐定(1 μM)诱导的荧光信号，IC50值为45 μM。由此数据计算出的抑制常数(Ki)为14 μM。[3] 研究指出，根据其他文献报道，在高浓度（1-100 μM）下，阿托品对α-肾上腺素能受体也具有拮抗活性。[3] 乙酰胆碱引起的人体肺静脉扩张可被氢溴酸阿托品（托品；1 μM；肺静脉和动脉）抑制 [4]。 |
| 体内研究 (In Vivo) |
文章讨论到，局部使用阿托品对儿童近视有效，但1%的剂量会引起副作用。近期研究表明，0.01%浓度的阿托品在保持抑制近视效果的同时，副作用有所减轻。[3]
在鸡的形觉剥夺性近视模型中，已知阿托品可以抑制近视，其在玻璃体中发挥作用的估计浓度范围为0.1-10 mM。[3] 论文引用了研究发现，即消融鸡的胆碱能无长突细胞并不会削弱阿托品对近视的抑制作用，这表明其作用部位可能不在视网膜。[3] 论文还提到，在鸡的视网膜-RPE-脉络膜-巩膜制剂中，使用抑制近视浓度的阿托品处理会导致视网膜神经递质大量、非特异性地释放。[3] 在小鼠巩膜成纤维细胞的体外制备物中，阿托品抑制卡巴胆碱诱导的细胞增殖作用，仅在高浓度（0.5-100 μM）下才能观察到，这比其对mAChRs的Ki值高出了500-1000倍。[3] 氢溴酸阿托品（peromyscus sp.；腹腔注射；10 mg/kg；40 分钟以上） 氢溴酸可抑制麻醉期间通常发生的心律失常[2]。 |
| 酶活实验 |
本研究未进行直接的酶学实验。主要使用的是一种基于细胞的CRE-荧光素酶报告基因实验来测量受体的激活和抑制。[3]
用于受体拮抗作用的CRE-荧光素酶实验： 将缺乏内源性M3受体的CRISPR-M3 HEK293T细胞，与受体克隆（人源M4、鸡源cM4或人源ADRA2A）、cAMP反应元件荧光素酶载体（CRE-Luc）和海肾荧光素酶对照载体（RLuc）共转染。转染48小时后，将细胞与固定亚最大浓度的激动剂（对于M4/cM4使用10 μM卡巴胆碱，对于ADRA2A使用1 μM可乐定）和递增浓度的待测拮抗剂（包括阿托品）一起孵育4小时。孵育后，裂解细胞，并使用Dual-Glo荧光素酶检测系统依次测量荧光素酶活性。CRE-Luc活性（反映cAMP水平和受体激活）被归一化到RLuc活性（作为细胞活力和转染效率的对照）。通过非线性回归分析归一化数据来确定拮抗剂的IC50值。[3] |
| 细胞实验 |
细胞培养和转染： CRISPR-M3 HEK293T细胞在含10% FBS的DMEM中培养。实验时，将细胞以30%的融合度接种在12孔板中，并使用Lipofectamine LTX进行转染。每个孔，将含有160 ng受体DNA（如人源M4）、180 ng CRE-Luc和160 ng RLuc的Opti-MEM混合物与Lipofectamine LTX溶液混合。室温孵育5分钟后，将复合物加入到细胞中。8小时后更换培养基，转染24小时后，将细胞消化并重新接种到白色底透的96孔板中，密度为每孔7500个细胞。[3]
CRE-荧光素酶发光实验： 在初次转染48小时后，吸去96孔板中的培养基，替换为50 μL含有固定浓度激动剂（M4/cM4用10 μM卡巴胆碱；ADRA2A用1 μM可乐定）和不同浓度拮抗剂（如阿托品）的FluoroBrite DMEM。细胞在37°C孵育4小时。随后，每孔加入50 μL Dual-Glo荧光素酶试剂。振荡孵育10分钟确保细胞裂解后，测量CRE-Luc发光信号。接着，每孔加入50 μL Dual-Glo Stop & Glo试剂，再次振荡孵育10分钟后，测量海肾荧光素酶发光信号。CRE-Luc值通过RLuc值进行归一化，以控制孔间差异。[3] |
| 动物实验 |
Animal/Disease Models: White-footed mice (Peromyscus sp.) [2]
Doses: 10 mg/kg Route of Administration: intraperitoneal (ip) injection; once, lasting 40 minutes. Experimental Results: increased heart rate and diminished arrhythmia. The paper discusses animal models and protocols from the perspective of reviewed literature, rather than presenting new in vivo data for atropine. [3] Chick Model of Myopia: In studies referenced by the paper, form-deprivation myopia (FDM) is induced in chicks. Atropine is administered to inhibit myopia, typically via intravitreal injection. Concentrations used range from 0.1 to 10 mM (estimated vitreal concentration), with a total amount of 20-2000 nmol per injection being common. [3] Rabbit Model for Ocular Distribution: The paper references studies where a single dose of 2% [3H]-atropine was delivered to the conjunctival sac of albino rabbits to study its distribution in ocular tissues. [3] Human Clinical Use: The paper discusses clinical protocols where atropine is delivered as daily eye drops at concentrations ranging from 0.01% to 1% for the treatment of childhood myopia. [3] |
| 药代性质 (ADME/PK) |
Ocular Distribution (from rabbit studies): One hour after a single topical dose of 2% [3H]-atropine to the conjunctival sac of albino rabbits, the concentration in ocular tissues was 0.09% of the original dose in the sclera, 0.05% in the choroid, and 0.008% in the retina. [3]
Another study cited found 0.01% in the retina and 0.38% in the choroid and sclera 30 minutes after topical application of [C14]-atropine. [3] Estimated Human Ocular Concentrations: Based on the rabbit data, a single topical dose of 0.01% to 1% atropine in humans is estimated to result in concentrations of 0.13-13 μM in the sclera, 0.07-7 μM in the choroid, and 0.01-1.1 μM in the retina. [3] Accumulation and Binding: The paper notes that serum levels of atropine can accumulate with repeated dosing, rising from undetectable levels after a single dose to 1.13-5.23 ng/μL when administered every 4-6 hours over 48 hours. [3] Furthermore, pigmented ocular tissues (RPE, iris, ciliary body, choroid, retina) are particularly adept at retaining atropine, which may lead to a prolonged effect. [3] |
| 毒性/毒理 (Toxicokinetics/TK) |
Clinical Side Effects: At the commonly prescribed 1% concentration for myopia treatment, atropine may induce allergic reactions and mAChR M3-mediated side effects such as mydriasis (pupil dilation), photophobia (light sensitivity), cycloplegia (loss of accommodation), and possibly early presbyopia. [3]
Rebound Effect: Upon cessation of 1% atropine treatment, a "rebound" phenomenon can occur, where myopia progresses at a faster rate than in untreated eyes. This is possibly due to desensitization of target receptors from the high drug concentration. [3] Side Effects at Low Dose: Even with the lower, more favored 0.01% concentration, some patients still complain of complications related to muscarinic receptor blockade, including photophobia and blurred vision. However, these symptoms are typically not severe enough to discontinue treatment. [3] In Vitro Cytotoxicity: In the cell-based assays of this study, high concentrations of atropine did not cause significant cell death in the CRISPR-M3 HEK293T cells, as determined by the Renilla luciferase control. [3] |
| 参考文献 | |
| 其他信息 |
Background: Atropine is a potent, non-selective muscarinic acetylcholine receptor (mAChR) antagonist. It has been the standard pharmacological treatment for myopia, but its mechanism of action in this context is unknown and is a central focus of the research paper. [3]
Mechanism of Action (Proposed): The paper hypothesizes that atropine's anti-myopia effects at the high concentrations used in vivo may be mediated through off-target binding to alpha2A-adrenoceptors (ADRA2A) rather than through its canonical target, the M4 mAChR. This is supported by the correlation between the drug's potency at hADRA2A and its reported ability to inhibit chick FDM, which is not seen for M4/cM4. [3] Clinical Use for Myopia: Topical atropine is effective in controlling myopia progression in children. Due to side effects at higher doses, 0.01% atropine eye drops are currently the most favored concentration prescribed by clinicians in Southeast Asia and North America. [3] Conclusion from the Study: The authors conclude from their data that the action of atropine via the mAChR M4 receptor is highly unlikely to be a factor in myopia control, and that non-mAChR targets, such as alpha-adrenoceptors, warrant further investigation. [3] |
| 分子式 |
C17H23NO3.HBR
|
|---|---|
| 分子量 |
370.28136
|
| 精确质量 |
369.094
|
| 元素分析 |
C, 55.14; H, 6.53; Br, 21.58; N, 3.78; O, 12.96
|
| CAS号 |
6415-90-3
|
| 相关CAS号 |
Atropine;51-55-8;Atropine sulfate;55-48-1
|
| PubChem CID |
21090485
|
| 外观&性状 |
Typically exists as solid at room temperature
|
| LogP |
2.826
|
| tPSA |
49.77
|
| 氢键供体(HBD)数目 |
2
|
| 氢键受体(HBA)数目 |
4
|
| 可旋转键数目(RBC) |
5
|
| 重原子数目 |
22
|
| 分子复杂度/Complexity |
353
|
| 定义原子立体中心数目 |
2
|
| SMILES |
Br.OCC(C(OC1CC2CCC(N2C)C1)=O)C1C=CC=CC=1
|
| InChi Key |
VZDNSFSBCMCXSK-ZZJGABIISA-N
|
| InChi Code |
InChI=1S/C17H23NO3.BrH/c1-18-13-7-8-14(18)10-15(9-13)21-17(20)16(11-19)12-5-3-2-4-6-12;/h2-6,13-16,19H,7-11H2,1H3;1H/t13-,14+,15?,16?;
|
| 化学名 |
[(1R,5S)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl] 3-hydroxy-2-phenylpropanoate;hydrobromide
|
| 别名 |
Atropine hydrobromide; 6415-90-3; Atropine (hydrobromide); 6VKW7R97ZU; 1alphaH,5alphaH-Tropan-3alpha-ol (+-)-tripate (ester), hydrobromide;
|
| HS Tariff Code |
2934.99.9001
|
| 存储方式 |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| 运输条件 |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| 溶解度 (体外实验) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| 溶解度 (体内实验) |
注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。
注射用配方
注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)(IP/IV/IM/SC等) *生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。 注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline) 注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil) 示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。 View More
注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)] 口服配方
口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠) 口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素) 示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。 View More
口服配方 3: 溶解于 PEG400 (聚乙二醇400) 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。 |
| 制备储备液 | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7007 mL | 13.5033 mL | 27.0066 mL | |
| 5 mM | 0.5401 mL | 2.7007 mL | 5.4013 mL | |
| 10 mM | 0.2701 mL | 1.3503 mL | 2.7007 mL |
1、根据实验需要选择合适的溶剂配制储备液 (母液)：对于大多数产品，InvivoChem推荐用DMSO配置母液 (比如：5、10、20mM或者10、20、50 mg/mL浓度），个别水溶性高的产品可直接溶于水。产品在DMSO 、水或其他溶剂中的具体溶解度详见上”溶解度 (体外)”部分;
2、如果您找不到您想要的溶解度信息，或者很难将产品溶解在溶液中，请联系我们;
3、建议使用下列计算器进行相关计算（摩尔浓度计算器、稀释计算器、分子量计算器、重组计算器等）;
4、母液配好之后，将其分装到常规用量，并储存在-20°C或-80°C，尽量减少反复冻融循环。
计算结果:
工作液浓度： mg/mL；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL)。如该浓度超过该批次药物DMSO溶解度，请首先与我们联系。
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL ddH2O，混匀澄清。
(1) 请确保溶液澄清之后，再加入下一种溶剂 (助溶剂) 。可利用涡旋、超声或水浴加热等方法助溶；
(2) 一定要按顺序加入溶剂 (助溶剂) 。
InvivoChem的所有产品仅用于作科学研究，不面向患者销售
Copyright 2020 InvivoChem LLC | All Rights Reserved 粤ICP备20063088号-1
粤公网安备 44011102003439号
463611831
