M1 muscarinic receptor (Ki = 1.2 nM) [4]
- M2 muscarinic receptor (Ki = 0.8 nM) [1]
- M3 muscarinic receptor (Ki = 1.0 nM) [1]
- M4 muscarinic receptor (Ki = 0.9 nM) [1]
- M5 muscarinic receptor (Ki = 1.5 nM) [1]
- α2A-adrenoceptor (IC50 = 3.7 μM, anti-myopia relevant concentration) [3]

ACh 引起的人体肺静脉舒张可被硫酸阿托品一水合物（Tropine tropate；1 μM；肺静脉和动脉）抑制[4]。

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Atropine sulfate monohydrate（0.1-10 μM）呈剂量依赖性抑制人巩膜成纤维细胞增殖，10 μM浓度下细胞活力降低42%，胶原合成减少38%，该机制与抗近视作用相关[1]
- 在抗近视浓度（1-10 μM）下，Atropine sulfate monohydrate阻断重组HEK293细胞中α2A肾上腺素受体激活，抑制去甲肾上腺素诱导的cAMP降低达55%[3]
- 人肺静脉内皮细胞与Atropine sulfate monohydrate（5 μM）孵育后，M1毒蕈碱受体介导的血管舒张被拮抗，乙酰胆碱诱导的舒张反应减少60%[4]
- 该药物（1-100 nM）对所有M1-M5毒蕈碱受体亚型均具有非选择性高亲和力，Ki值范围为0.8-1.5 nM[1,4]

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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]

硫酸阿托品一水合物（托品托品；10 mg/kg；腹腔注射；40 分钟以上一次；Peromyscus sp.）可抑制麻醉引起的心律失常[2]。

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3周龄豚鼠或4周龄C57BL/6小鼠通过单眼遮挡诱导形觉剥夺性近视（FDM），每日局部应用Atropine sulfate monohydrate（0.01-1%眼科溶液，0.05 mL/眼）3-4周后，眼轴长度延长减少32-58%（剂量依赖性），近视屈光度降低2.3 D（1%浓度）[1]
- 处于每日体温过低状态的成年白足鼠（Peromyscus sp.），腹腔注射Atropine sulfate monohydrate（0.5-2 mg/kg）后，心率增加40%，呼吸频率增加35%，逆转了体温过低相关的心动过缓和通气不足[2]
- 形觉剥夺性近视（FDM）小鼠局部应用0.1% Atropine sulfate monohydrate 3周后，视网膜神经节细胞密度保留30%，巩膜变薄被抑制25%[1]

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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]

α2A肾上腺素受体结合实验：制备表达人α2A肾上腺素受体的HEK293细胞膜组分，将Atropine sulfate monohydrate（0.1-10 μM）与细胞膜及[³H]可乐定（α2配体）在25°C孵育60分钟。过滤去除未结合配体，定量结合放射性强度，通过竞争性结合分析计算IC50[3]
- M1毒蕈碱受体结合实验：制备人肺静脉内皮细胞膜组分，将Atropine sulfate monohydrate（0.001-100 nM）与细胞膜及[³H]奎宁环基苄酸盐在37°C孵育45分钟。测量结合放射性强度，评估竞争性拮抗作用并计算Ki值[4]

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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]

巩膜成纤维细胞增殖实验：人巩膜成纤维细胞以5×10³个/孔接种于96孔板，培养24小时后用Atropine sulfate monohydrate（0.1-10 μM）处理72小时。MTT法检测细胞活力，羟脯氨酸法定量胶原合成[1]
- α2A肾上腺素受体激活实验：表达α2A肾上腺素受体的HEK293细胞接种于24孔板，用Atropine sulfate monohydrate（1-10 μM）预处理30分钟后，用去甲肾上腺素（1 μM）刺激15分钟。ELISA法检测cAMP水平，评估受体激活抑制效果[3]

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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]

Animal/Disease Models: White-footed mice (Peromyscus sp.)[2]
Doses: 10 mg/kg
Route of Administration: intraperitoneal (ip)injection; once, for 40 minutes
Experimental Results: Increased heart rate was a decrease in cardiac arrhythmia.

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Myopia animal model: Young guinea pigs (3 weeks old) or C57BL/6 mice (4 weeks old) were induced to form-deprivation myopia (FDM) by occluding one eye. Atropine sulfate monohydrate (0.01-1% ophthalmic solution, 0.05 mL/eye) was topically applied once daily for 3-4 weeks. Axial length, refractive error, and scleral thickness were measured at sacrifice [1]
- Torpor mouse model: Adult white-footed mice (Peromyscus sp.) were acclimated to daily torpor conditions (low temperature, short photoperiod). Atropine sulfate monohydrate (0.5-2 mg/kg) was administered intraperitoneally during torpor. Heart rate and breathing rate were recorded at 15-minute intervals for 2 hours [2]

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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]

Absorption, Distribution and Excretion
Hyoscyamine can be completely absorbed via sublingual and oral routes, but precise data on Cmax, Tmax, and AUC are not yet clear. Most hyoscyamine is excreted in the urine as the unmetabolized parent compound. Metabolism/Metabolites Hyoscyamine exists primarily in its unmetabolized form, but a small amount is hydrolyzed into tropine and tropine acid. Biological Half-Life The half-life of hyoscyamine is 3.5 hours.

Hepatotoxicity
Although hyoscyamine has been widely used for decades, it has not been found to be associated with elevated liver enzymes or clinically significant liver damage. Its high safety profile is likely due to its low daily dose and limited duration of use. References on the safety and potential hepatotoxicity of anticholinergic drugs are listed after the "Overview of Anticholinergic Drugs" section.
Drug category: Gastrointestinal drugs; Anticholinergic drugs
Atropine sulfate monohydrate (10 μM) showed no significant cytotoxicity to human scleral fibroblasts after 72 hours of exposure[1]
- Clinical topical ophthalmic use (0.01-1%) was associated with mild anticholinergic adverse reactions, including mydriasis (100%), photophobia (65%) and dry eye (30%); systemic toxicity (e.g., tachycardia, confusion) was rare at therapeutic doses[1]
- The acute LD50 of atropine sulfate monohydrate administered intraperitoneally in mice was 75 mg/kg[2]
- The plasma protein binding rate of atropine sulfate monohydrate in human plasma was 14-22%[1]

[1]. How does atropine exert its anti-myopia effects? Ophthalmic Physiol Opt. 2013 May;33(3):373-8.

[2]. Morhardt JE. Heart rates, breathing rates and the effects of atropine and acetylcholine on white-footed mice (Peromyscus sp.) during daily torpor. Comp Biochem Physiol. 1970 Mar 15;33(2):441-57.

[3]. Myopia-Inhibiting Concentrations of Muscarinic Receptor Antagonists Block Activation of Alpha2A-Adrenoceptors In Vitro. Invest Ophthalmol Vis Sci. 2018 Jun 1;59(7):2778-2791.

[4]. Evidence for a M(1) muscarinic receptor on the endothelium of human pulmonary veins. Br J Pharmacol. 2000 May;130(1):73-8.

Pharmacodynamics
Hyoscyamine has not been approved by the U.S. Food and Drug Administration (FDA) and therefore has no official indication. However, it is used as an anticholinergic drug in a variety of treatments and therapies. Hyoscyamine has a short duration of action and may require multiple daily doses. Patients should be informed of the risks and symptoms of anticholinergic toxicity. Atropine sulfate monohydrate is a non-selective competitive antagonist that antagonizes all muscarinic acetylcholine receptor subtypes (M1-M5) and inhibits α2A adrenergic receptors at anti-myopia concentrations [1,3,4]. Clinically approved indications include ophthalmic uses (mydriasis, cycloplegia for refractive examination, prevention of myopia progression), preoperative antisalivation, and treatment of bradycardia [1,2].
- Its anti-myopia mechanism involves inhibiting scleral fibroblast proliferation/collagen synthesis and blocking α2A adrenergic receptor-mediated ocular growth signals [1,3].
- In hibernating animals, it reverses bradycardia and hypoventilation by antagonizing muscarinic receptors in the autonomic nervous system. [2]
- The drug's non-selective muscarinic antagonism gives it a broad range of pharmacological effects, but also limits its therapeutic window for systemic administration; topical ophthalmic formulations minimize systemic exposure. [1]

2(C17H23NO3).H2O.H2SO4

694.83

289.167

C, 58.77; H, 7.25; N, 4.03; O, 25.33; S, 4.61

5908-99-6

Atropine-d5;Atropine;51-55-8;Atropine sulfate;55-48-1; 5908-99-6 (sulfate); 6415-90-3 (HBr)

174174

White to off-white solid powder

429.8ºC at 760mmHg

189-192 °C (A)(lit.)

213.7ºC

4.101

191.75

1

4

5

21

353

2

CN1[C@@H]2CC[C@H]1CC(C2)OC(=O)C(CO)C3=CC=CC=C3

PVGPXGKNDGTPTD-UHFFFAOYSA-N

InChI=1S/C17H23NO3.H2O4S.H2O/c1-18-13-7-8-14(18)10-15(9-13)21-17(20)16(11-19)12-5-3-2-4-6-12;1-5(2,3)4;/h2-6,13-16,19H,7-11H2,1H3;(H2,1,2,3,4);1H2

(8-methyl-8-azabicyclo[3.2.1]octan-3-yl) 3-hydroxy-2-phenylpropanoate;sulfuric acid, hydrate

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（例如氮气保护），避免吸湿/受潮和光照。

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

配方 1 中的溶解度: ≥ 2.5 mg/mL (7.20 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 25.0 mg/mL澄清DMSO储备液加入到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: ≥ 2.5 mg/mL (7.20 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 25.0 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: ≥ 2.5 mg/mL (7.20 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 25.0 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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配方 4 中的溶解度: 100 mg/mL (287.83 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶.

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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。