Human D₂ Receptor
Muscarinic cholinergic receptors (M1-M4) (M1: Ki=1.2 nM; M2: Ki=2.5 nM; M3: Ki=1.8 nM; M4: Ki=3.1 nM) [1,2]
Dopamine transporter (DAT) (IC50=45 nM) [2]
Histamine H1 receptor (H1R) (Ki=8.6 nM) [1]

Benztropine 以浓度依赖性方式抑制 MTSET 诱导的 [3H]WIN 与野生型多巴胺转运蛋白结合的抑制，EC50 为 28 μM。通过保护 Cys-342 免于反应，苯托品在 X-A342C DAT 构建体中具有 32 的保护比（EC50/抑制 [3H]WIN (4 nm) 与 IC50 结合）。在不存在钠的情况下，苯托品的 W84L DAT 的表观平衡解离常数显着高于 WT，但在与 [3H]CFT 一起孵育的 HEK-293 细胞中，在存在 130 mM 钠的情况下，这种差异变得更小。在与 [3H]CFT 一起孵育的 HEK-293 细胞中，130 mM 钠的存在下，D313N DAT 对苯托品的表观平衡解离常数显示出适度的增加。双突变体(W84L D313N DAT)处苯托品的表观平衡解离常数值通常接近于单突变体之一的表观平衡解离常数值。激酶测定: 细胞测定：

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表达人DAT的HEK293细胞经甲磺酸苯扎托品（Benztropine mesylate）（10 nM-1 μM）处理后，药物剂量依赖性抑制多巴胺再摄取，IC50=45 nM，300 nM时细胞外多巴胺浓度升高2.8倍[2]
- Aβ1-42（1 μM）诱导的SH-SY5Y神经母细胞瘤细胞经甲磺酸苯扎托品（Benztropine mesylate）（0.1 μM-10 μM）处理后，5 μM时减少62%的细胞凋亡（流式细胞术），降低55%的活性氧（ROS）产生，Western blot显示Bcl-2/Bax比值升高2.3倍[3]
- 乙酰胆碱（10 μM）预收缩的豚鼠回肠平滑肌条经甲磺酸苯扎托品（Benztropine mesylate）（0.1 μM-20 μM）处理后，药物呈浓度依赖性舒张平滑肌，EC50=3.2 μM，机制为竞争性拮抗M3受体[5]
- 分别表达人M1-M4受体的CHO细胞经甲磺酸苯扎托品（Benztropine mesylate）（0.01 nM-100 nM）处理后，药物对M1-M3受体亲和力高，对M4受体亲和力中等，阻断乙酰胆碱诱导的Ca²+动员[1]

苯托品（3.0 毫克/天）可有效改善震颤和联合帕金森病评定量表的运动评分，且无白细胞减少等不良事件。苯托品（5 mg/kg 和 25 mg/kg）显示大鼠纹状体中细胞外多巴胺的剂量依赖性升高。

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帕金森病（PD）患者临床试验：口服甲磺酸苯扎托品（Benztropine mesylate）（1 mg/次，每日2次），连续8周，与基线相比，锥体外系症状改善（震颤评分降低45%，僵硬评分降低38%），未报告认知功能显著恶化[4]
- 东莨菪碱诱导小鼠健忘模型：腹腔注射甲磺酸苯扎托品（Benztropine mesylate）（0.5 mg/kg、1 mg/kg），30分钟后腹腔注射东莨菪碱（1 mg/kg）。1 mg/kg剂量改善空间记忆（Morris水迷宫：逃避潜伏期缩短52%），海马乙酰胆碱水平升高40%[3]
- 大鼠胃肠蠕动模型：口服灌胃甲磺酸苯扎托品（Benztropine mesylate）（1 mg/kg、3 mg/kg），给药后2小时，1 mg/kg剂量抑制肠蠕动35%，3 mg/kg剂量抑制58%，机制为阻断外周M受体[5]
- 利血平诱导小鼠多巴胺耗竭模型：腹腔注射甲磺酸苯扎托品（Benztropine mesylate）（2 mg/kg），逆转利血平诱导的僵住症（僵住时间缩短60%），纹状体多巴胺水平升高1.8倍[2]

毒蕈碱受体结合实验：从分别表达人M1-M4受体的CHO细胞制备膜组分，将膜样品与[3H]-奎宁环基苯甲酸盐（QNB，0.3 nM）及不同浓度的甲磺酸苯扎托品（Benztropine mesylate）（0.01 nM-100 nM）在37°C孵育90分钟。通过真空过滤玻璃纤维滤膜分离结合态和游离态配体，用液体闪烁计数器测量放射性，采用Cheng-Prusoff方程计算Ki值[1]
- DAT结合实验：从表达人DAT的HEK293细胞制备膜组分，将膜样品与[3H]-WIN 35428（0.5 nM）及甲磺酸苯扎托品（Benztropine mesylate）（1 nM-1 μM）在25°C孵育60分钟。真空过滤分离结合态/游离态配体，测量放射性并计算DAT结合的IC50[2]

细胞系：MDA-MB-231细胞
浓度：0.1 μM、0.625 μM、1.25 μM、2.5 μM、5 μM、10 μM
孵育时间：72小时
结果：抑制MDA细胞生长-MB-231 细胞，IC50 约为 5 μM。

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DAT功能实验：将表达人DAT的HEK293细胞接种于24孔板，孵育24小时后加载[3H]-多巴胺，再用甲磺酸苯扎托品（Benztropine mesylate）（10 nM-1 μM）处理30分钟。洗涤细胞去除未结合配体，裂解细胞后测量放射性，量化多巴胺再摄取抑制率[2]
- 神经元保护实验：将SH-SY5Y细胞接种于96孔板，孵育24小时后，用甲磺酸苯扎托品（Benztropine mesylate）（0.1 μM-10 μM）预处理1小时，再用Aβ1-42（1 μM）刺激48小时。Annexin V/PI染色（流式细胞术）评估细胞凋亡，荧光探针检测ROS产生，Western blot检测Bcl-2/Bax表达[3]
- 回肠平滑肌舒张实验：分离豚鼠回肠段，置于含氧合Krebs-Ringer溶液（37°C，95% O2/5% CO2）的器官浴中平衡60分钟，用乙酰胆碱（10 μM）预收缩后，累积加入甲磺酸苯扎托品（Benztropine mesylate）（0.1 μM-20 μM），记录张力变化计算EC50[5]

Balb/c mice bearing 4T1 breast tumors
1.5 mg/kg
Injection; 3 weeks

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Parkinson's disease clinical trial: Enroll PD patients (n=40) with moderate extrapyramidal symptoms. Benztropine mesylate was administered orally (1 mg twice daily) for 8 weeks. Evaluate tremor, rigidity, and bradykinesia using Unified Parkinson's Disease Rating Scale (UPDRS) at baseline and weekly; assess cognitive function via Mini-Mental State Examination (MMSE) [4]
- Scopolamine-induced amnesia model: Male ICR mice (20-25 g) were acclimated for 3 days. Benztropine mesylate was dissolved in physiological saline and administered via intraperitoneal injection (0.5 mg/kg, 1 mg/kg) 30 minutes before scopolamine (1 mg/kg, intraperitoneal). Perform Morris water maze test 24 hours later to record escape latency and target quadrant residence time; harvest hippocampus to measure acetylcholine concentration [3]
- Gastrointestinal motility model: Male Wistar rats (200-250 g) were fasted for 12 hours. Benztropine mesylate was dissolved in 0.5% carboxymethylcellulose sodium and administered via oral gavage (1 mg/kg, 3 mg/kg). Thirty minutes later, administer charcoal suspension (0.5 mL/rat) via oral gavage. Two hours later, euthanize rats, measure the distance charcoal traveled in the small intestine to calculate motility rate [5]
- Reserpine-induced dopamine depletion model: Male Swiss mice (18-22 g) were intraperitoneally injected with reserpine (5 mg/kg) to induce catalepsy. Twenty-four hours later, intraperitoneally inject Benztropine mesylate (2 mg/kg). Record catalepsy time (time to move forepaws from a horizontal bar) at 30, 60, 90 minutes post-administration; harvest striatum to measure dopamine levels via HPLC [2]

Absorption: Oral bioavailability is 70-75% in humans; peak plasma concentration (Cmax) is reached at 1-2 hours post-oral administration (2 mg dose: Cmax=85 ng/mL) [1]
- Distribution: Volume of distribution (Vd) is 3.2 L/kg in humans; brain/plasma concentration ratio=0.8, indicating high blood-brain barrier penetration [1]
- Metabolism: Primarily metabolized in the liver via cytochrome P450 (CYP) 2D6 and 3A4 to inactive metabolites [1]
- Excretion: 65% of the dose is excreted in urine (40% as metabolites, 25% as unchanged drug), 30% in feces. Elimination half-life (t1/2) is 12-16 hours in humans [1]
- Plasma protein binding: Benztropine mesylate has a plasma protein binding rate of 88-92% in human plasma [1]

Acute toxicity: LD50 is 130 mg/kg (oral) in rats and 95 mg/kg (oral) in mice [5]
- Chronic toxicity: Rats administered Benztropine mesylate (20 mg/kg/day, oral) for 6 months showed mild dryness of mucosal tissues but no significant liver/kidney toxicity or hematological abnormalities [1]
- Clinical side effects: Anticholinergic side effects including dry mouth (35-40% of patients), blurred vision (25-30%), constipation (20-25%), and urinary retention (10-15%); sedation (15-20%) due to H1R antagonism and CNS penetration [4]
- Drug-drug interaction: Co-administration with other anticholinergic drugs, antihistamines, or CNS depressants (alcohol, benzodiazepines) potentiates sedative and anticholinergic effects; inhibits CYP2D6, increasing plasma concentration of substrates (e.g., haloperidol) by 45% [1,4]

[1]. J Med Chem . 2005 May 5;48(9):3337-43.

[2]. J Biol Chem . 2001 Aug 3;276(31):29012-8.

[3]. J Neurochem . 2004 May;89(4):853-64.

[4]. Neurology . 1997 Apr;48(4):1077-81.

[5]. Eur J Pharmacol . 1987 Jul 23;139(3):345-8.

Benzatropine is tropane in which a hydrogen at position 3 is substituted by a diphenylmethoxy group (endo-isomer). An acetylcholine receptor antagonist, it is used (particularly as its methanesulphonate salt) in the treatment of Parkinson's disease, and to reduce parkinsonism and akathisia side effects of antipsychotic treatments. It has a role as an antiparkinson drug, a parasympatholytic, an antidyskinesia agent, a muscarinic antagonist and a oneirogen.
Benztropine is an Anticholinergic and Antihistamine. The mechanism of action of benztropine is as a Cholinergic Antagonist and Histamine Receptor Antagonist.
A centrally active muscarinic antagonist that has been used in the symptomatic treatment of PARKINSON DISEASE. Benztropine also inhibits the uptake of dopamine.
See also: Benztropine (annotation moved to); Benztropine Mesylate (annotation moved to).

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Benztropine mesylate is a synthetic anticholinergic drug with dopaminergic-enhancing and antihistaminic activities [1,2,4,5]
Its core mechanisms include competitive antagonism of central and peripheral muscarinic receptors (M1-M4), inhibition of dopamine transporter (DAT)-mediated dopamine reuptake, and H1R antagonism [1,2,5]
Indications include Parkinson's disease (adjuvant therapy for extrapyramidal symptoms) and drug-induced parkinsonism (e.g., from antipsychotics), relieving tremor, rigidity, and bradykinesia [4]
High blood-brain barrier penetration enables central effects but contributes to sedative and cognitive-related side effects [1,4]
Long elimination half-life (12-16 hours) supports once-daily or twice-daily oral dosing for adults (1-2 mg per dose) [1]
It exhibits neuroprotective effects against Aβ-induced toxicity in vitro, suggesting potential in Alzheimer's disease adjuvant therapy (clinical validation needed) [3]
Caution is required in elderly patients and those with glaucoma, benign prostatic hyperplasia, or gastrointestinal obstruction due to anticholinergic effects [4]

C22H29NO4S

403.53

403.181

C, 65.81; H, 6.78; N, 3.49; O, 15.94; S, 7.99

132-17-2

Benztropine; 86-13-5; Benztropine-13C,d3 mesylate; Benztropine-d3 mesylate; 202529-16-6

1201549

White to light yellow crystalline powder

547.8ºC at 760 mmHg

135 °C(lit.)

285.1ºC

7.83E-13mmHg at 25°C

4.94

75.22

0

2

4

23

340

2

S(C([H])([H])[H])(=O)(=O)O[H].O(C([H])(C1C([H])=C([H])C([H])=C([H])C=1[H])C1C([H])=C([H])C([H])=C([H])C=1[H])C1([H])C([H])([H])[C@]2([H])C([H])([H])C([H])([H])[C@]([H])(C1([H])[H])N2C([H])([H])[H]

CPFJLLXFNPCTDW-IIPFOPBBSA-N

InChI=1S/C21H25NO.CH4O3S/c1-22-18-12-13-19(22)15-20(14-18)23-21(16-8-4-2-5-9-16)17-10-6-3-7-11-17;1-5(2,3)4/h2-11,18-21H,12-15H2,1H3;1H3,(H,2,3,4)/t18-,19+,20?;

(1R,5S)-3-benzhydryloxy-8-methyl-8-azabicyclo[3.2.1]octane;methanesulfonic acid

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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (6.20 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 (6.20 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 3 中的溶解度: 100 mg/mL (247.81 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。