NSC305787 is an inhibitor of ezrin with a K_d of 5.85 μM, inhibits the phosphorylation of ezrin caused by PKCΙ with an IC₅₀ of 8.3 μM, has antitumor activity.

Kd: 5.85 μM (ezrin)^[1]
IC50: 8.3 μM (ezrin)^[1]

NSC305787 is an inhibitor of ezrin with a K_d of 5.85 μM, and has antitumor activity. NSC305787 inhibits PKCΙ phosphorylation of Ezrin, Moesin, Radixin, MBP, with IC₅₀s of 8.3, 9.4, 55, 58.9 μM, respectively. NSC305787 binds to PKCΙ with a K_d value of 172.4 μM, and inhibits ezrin T567 phosphorylation primarily via its binding to ezrin and not through inhibition of PKCΙ kinase activity. NSC305787 (1, 10 μM) shows inhibitory activity against ezrin-mediated invasion of K7M2 osteosarcoma (OS) cells. Moreover, NSC305787 (10 μM) reduces cell motility phenotypes in zebrafish and blocks OS metastatic growth in lung organ culture^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

NSC305787 (0.240 mg/kg/day, i.p.) suppresses ezrin-dependent osteosarcoma metastatic growth in mouse lung^[1]. NSC305787 (240 μg/kg, i.p.) dramatically inhibits pulmonary metastasis in a transgenic mouse model of osteosarcoma (Osx-Cre⁺p53^fl/flpRB^fl/fl) and shows a more favorable pharmacokinetic profile compared with NSC668394 in the mouse model^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

445.42

C₂₅H₃₀Cl₂N₂O

785718-37-8

Room temperature in continental US; may vary elsewhere.

DMSO : 6 mg/mL (13.47 mM; Need ultrasonic and warming)

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。

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: ≥ 0.6 mg/mL (1.35 mM); Clear solution

  此方案可获得 ≥ 0.6 mg/mL (1.35 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 6.0 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: ≥ 0.6 mg/mL (1.35 mM); Clear solution

  此方案可获得 ≥ 0.6 mg/mL (1.35 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 6.0 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: ≥ 0.6 mg/mL (1.35 mM); Clear solution

  此方案可获得 ≥ 0.6 mg/mL (1.35 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 6.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Bulut G, et al. Small molecule inhibitors of ezrin inhibit the invasive phenotype of osteosarcoma cells. Oncogene. 2012 Jan 19;31(3):269-81.

  [2]. Çelik H, et al. Ezrin Inhibition Up-regulates Stress Response Gene Expression. J Biol Chem. 2016 Jun 17;291(25):13257-70.

Human umbilical vein endothelial cells (HUVECs; 2.5 × 10⁴/well) are seeded in a 96-well plate in endothelial growth media-2. Following formation of a confluent HUVEC monolayer (∼32 h), endothelial growth media-2 is aspirated and a layer of osteosarcoma (OS) cells (1.0 × 10⁴ cells/well) is added to Dulbecco’s modified Eagle medium containing NSC305787. This specific time point is accepted as 0 h of treatment, and invasion is monitored during the subsequent 5 h by measuring changes in resistance at the cell-electrode interphase. The cell index is calculated according to the following formula: cell index = (Rt – R0)/F, where Rt is resistance at time point t, R0 is background resistance (measured with media alone, no cells) and F is frequency at which the measurement is taken (10 kHz)^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[2]
The Osx-Cre⁺p53^fl/flpRB^fl/fl transgenic mouse model of osteosarcoma is used in the assay. Mice are treated with 240 μg/kg/day NSC305787, 226 μg/kg/day NSC668394, or vehicle (DMSO, 1%) once daily, five times a week by i.p. injection in a volume of 100 μL. At the end of the study, lung and tumor samples are isolated upon necropsy. Half of each sample is flash frozen immediately in liquid nitrogen, and the other half is fixed in 10% formalin for 18-24 h, transferred to 70% ethanol, and stored at room temperature^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Bulut G, et al. Small molecule inhibitors of ezrin inhibit the invasive phenotype of osteosarcoma cells. Oncogene. 2012 Jan 19;31(3):269-81.

  [2]. Çelik H, et al. Ezrin Inhibition Up-regulates Stress Response Gene Expression. J Biol Chem. 2016 Jun 17;291(25):13257-70.

NSC668394 is a potent ezrin (Thr567) phosphorylation inhibitor, with a K_d of 12.59 μM. NSC668394 inhibit ezrin T567 phosphorylation caused by PKCΙ primarily via their binding to ezrin. NSC668394 can be used to prevent tumor metastasis^[1][2][3].

NSC668394 (10 μM; pretreated for 15 min) inhibits ezrin T567 phosphorylation (IC₅₀=8.1 μM) and actin binding in vitro^[1].
NSC668394 (1-10 μM; 2-6 h) inhibits ezrin-mediated invasion by K7M2 osteosarcoma (OS) cells on the HUVEC monolayer^[1].
NSC668394 (20 μM) causes significant decrease in growth in JM1 and JM2 rat hepatoma cell lines^[2].
NSC668394 (10 μM) reduces cell motility phenotypes in zebrafish^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

NSC668394 (0.226 mg/kg/day; i.p. 5-days a week) inhibits ezrin-dependent in vivo OS metastatic growth in mouse lung^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

452.10

C₁₇H₁₂Br₂N₂O₃

382605-72-3

Room temperature in continental US; may vary elsewhere.

DMSO : 3.85 mg/mL (8.52 mM; ultrasonic and warming and heat to 80°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。-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。

• [1]. Bulut G, et, al. Small molecule inhibitors of ezrin inhibit the invasive phenotype of osteosarcoma cells. Oncogene. 2012 Jan 19;31(3):269-81.

  [2]. Xue Y, et, al. Phosphorylated Ezrin (Thr567) Regulates Hippo Pathway and Yes-Associated Protein (Yap) in Liver. Am J Pathol. 2020 Jul;190(7):1427-1437.

  [3]. Çelik H, et, al. Ezrin Inhibition Up-regulates Stress Response Gene Expression. J Biol Chem. 2016 Jun 17;291(25):13257-70.