| Description |
NSC305787 hydrochloride is an inhibitor of ezrin with a Kd of 5.85 μM, inhibits the phosphorylation of ezrin caused by PKCΙ with an IC50 of 8.3 μM, has antitumor activity.
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| Related Catalog |
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| Target |
Kd: 5.85 μM (ezrin)[1] IC50: 8.3 μM (ezrin)[1]
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| In Vitro |
NSC305787 hydrochloride is an inhibitor of ezrin with a Kd 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 Kd 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].
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| In Vivo |
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+p53fl/flpRBfl/fl) and shows a more favorable pharmacokinetic profile compared with NSC668394 in the mouse model[2].
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| Cell Assay |
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].
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| Animal Admin |
Mice[2] The Osx-Cre+p53fl/flpRBfl/fl transgenic mouse model of osteosarcoma is used in the assay. Female mice are fed with doxycycline-containing food (2,000 mg/kg diet) throughout the pregnancy until the weaning of the offspring at 21 days to repress the expression of Cre recombinase. All of the animals are observed daily, and tumors are measured at least once a week using a slide caliper. The tumor volumes are calculated using the following formula: tumor volume = (π/6) × length2 × width. Once tumors are detected, mice are randomly assigned to control (DMSO), NSC305787 treatment, and NSC668394 treatment groups. 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].
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| References |
[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.
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