Scopoletin is an inhibitor of acetylcholinesterase (AChE).

AChE^[1]

Scopoletin (SCT) is identified as a putative inhibitor of acetylcholinesterase (AChE). Scopoletin enhances the K⁺-stimulated release of ACh from rat frontal cortex synaptosomes, showing a bell-shaped dose effect curve (E_max: 4 μM) ^[1]. Scopoletin inhibits PC3 proliferation by inducing apoptosis of PC3 cells. The IC50 of Scopoletin for inhibiting PC3, PAA (human lung cancer cell), and Hela cell proliferation is (157±25), (154±51), and (294±100) mg/L, respectively. Scopoletin induces a marked time- and concentration-dependent inhibition of PC3 cell proliferation. Scopoletin reduces the protein content and decreases the acid phosphatase activity (ACP) level in PC3 cells in a concentration-dependent manner. Cells treated by Scopoletin show typical morphologic changes of apoptosis by light microscope, fluorescence microscope, and transmission electronmicroscope. Apoptosis rate is 0.3 %, 2.1 %, 9.3 % and 35 % for Scopoletin 0, 100, 200, and 400 mg/L, respectively, and cells in G2 phase decrease markedly after being treated with Scopoletin^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Scopoletin (2 μg, i.c.v.) increases T-maze alternation and ameliorated novel object recognition of mice with scopolamine-induced cholinergic deficit. It also reduces age-associated deficits in object memory of 15-18-month-old mice (2 mg/kg sc). Mice injected with 2 μg Scopoletin show an increased alternation rate of 71.3±2.5%^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

192.17

C₁₀H₈O₄

92-61-5

东莨菪内酯；莨菪亭；东莨菪亭

Room temperature in continental US; may vary elsewhere.

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

In Vitro: 

DMSO : 62.5 mg/mL (325.23 mM; Need ultrasonic)

Ethanol : < 1 mg/mL (ultrasonic) (insoluble)

*Scopoletin is usually formulated as a suspension.

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.08 mg/mL (10.82 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (10.82 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.08 mg/mL (10.82 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (10.82 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 3.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.08 mg/mL (10.82 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (10.82 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Hornick A, et al. The coumarin Scopoletin potentiates acetylcholine release from synaptosomes, amplifies hippocampal long-term potentiation and ameliorates anticholinergic- and age-impaired memory. Neuroscience. 2011 Dec 1;197:280-92.

  [2]. Liu XL, et al. Effect of Scopoletin on PC3 cell proliferation and apoptosis. Acta Pharmacol Sin. 2001 Oct;22(10):929-33.

PC3 cells (5×10⁷/L) 1 mL in exponential growth are seeded into four 24-well plates. The plates are incubated at 37°C in a humidified 5% CO₂ atmospbee. After 24h, Scopoletin 33, 66, 133, 266, and 533 mg/L are added to wells (3 wells for each concentration for each plate). For control cells (3 wells foreach plate),only DMEM was added. The plates are incubated continually. The viable cells are counted by hemocytometer every day in the frist 4 d by Trypan blue dye exclusion method^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[1]
C56BL/6N male, 4-6-month-old and 16-18-month-old mice are used in the behavioral studies. The mice are housed in groups of four in cages at constant humidity (50-55%) and temperature (22±1°C) on a 12:12 h light/dark cycle (7:00–19:00 h), with food and water ad libitum. Younger mice (4-6 months) are implanted with i.c.v. cannulas for application of Scopolamine (SCOP) and Scopoletin. The aged mice are injected with Scopoletin by the s.c. route. Experiments are conducted between 8:00 and 16:00 h. Mice with i.c.v. cannulas are randomly divided into four experimental groups: vehicle; SCOP 20 μg; Scopoletin 2 μg; and SCOP 20 μg plus Scopoletin 2 μg. The drugs are applied in 1 μL of vehicle solution (SCOP: saline, Scopoletin: 3 DMSO: 7 sterile water). I.c.v. injections are carried out 15 min before the start of the tests. Aged mice obtained Scopoletin s.c. 30 min prior to object memory test (vehicle: 1 DMSO: 1 EtOH, diluted with olive oil as required)^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Hornick A, et al. The coumarin Scopoletin potentiates acetylcholine release from synaptosomes, amplifies hippocampal long-term potentiation and ameliorates anticholinergic- and age-impaired memory. Neuroscience. 2011 Dec 1;197:280-92.

  [2]. Liu XL, et al. Effect of Scopoletin on PC3 cell proliferation and apoptosis. Acta Pharmacol Sin. 2001 Oct;22(10):929-33.