Digoxin is a potent inhibitor of Na+/K+-ATPase, has the potential for arrhythmia and heart failure treatment.

Digoxin, a sodium-potassium ATPase inhibitor, has a negative Z score of −26.67, suggesting a function for the sodium-potassium ATPase in CHIKV infection. Relative to DMSO-treated cells, treatment of U-2 OS cells with Digoxin results in a dose-dependent decrease in CHIKV infection with a half-maximal effective concentration (EC₅₀) of 48.8 nM. Digoxin treatment similarly decreases CHIKV infection of primary human synovial fibroblasts (HSFs) and Vero African green monkey kidney cells with EC₅₀s of 43.9 nM and 67.3 nM, respectively. Digoxin treatment significantly diminishes CHIKV infection in these cell types, with EC₅₀s of 16.2 µM in ST2 cells and 23.2 µM in C2C12 cells, values 330 and 475 times the EC₅₀ of Digoxin in U-2 OS cells. Cell viability is only modestly impaired at 24 h posttreatment with 1 µM Digoxin, a dose 20 times the Digoxin EC₅₀ for CHIKV antiviral activity in these cells^[1].

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

780.94

C₄₁H₆₄O₁₄

20830-75-5

地高辛；异羟洋地黄毒苷

Room temperature in continental US; may vary elsewhere.

4deg;C, protect from light

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (protect from light)

DMSO : 62.5 mg/mL (80.03 mM; ultrasonic and warming and heat to 60°C)

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 (protect from light)。-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。

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 (2.66 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (2.66 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 (2.66 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (2.66 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 (2.66 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (2.66 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]. Ashbrook AW, et al. Antagonism of the Sodium-Potassium ATPase Impairs Chikungunya Virus Infection. MBio. 2016 May 24;7(3). pii: e00693-16.

U-2 OS cells seeded in 60-mm-diameter dishes are incubated with DMSO, 10 µM STS as an inducer of apoptosis, or increasing concentrations of Digoxin at 37°C for 6 h. Cells are washed with fluorescence-activated cell sorter (FACS) buffer (PBS with 2% FBS) and stained with PI. Cell staining is quantified using a BD LSRII flow cytometer and FlowJo software. Alternatively, U-2 OS cells seeded in 96-well plates are incubated with DMSO, STS, or increasing concentrations of Digoxin at 37°C for 6 or 24 h. PrestoBlue reagent is added to supernatants of compound-treated cells, and cells are incubated at 37°C for 30 min. Fluorescence as a surrogate for cell viability is quantified using a Synergy H1 plate reader^[1].

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

• [1]. Ashbrook AW, et al. Antagonism of the Sodium-Potassium ATPase Impairs Chikungunya Virus Infection. MBio. 2016 May 24;7(3). pii: e00693-16.