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Rafoxanide is a novel allosteric inhibitor of SPAK and OSR1. Rafoxanide is also a salicylanilide used as an anthelmintic.
In vitro kinase assays using Rafoxanide showed it to be a low micromolar inhibitor of OSR1 T185E in vitro, IC50 = 8.18 μM. Rafoxanide is also a low micromolar inhibitor of SPAK T233E in vitro with IC50 value of 13.03 μM.
| Cell Experiment | |
|---|---|
| Cell lines | HEK293 cells |
| Preparation method | HEK293 cells were first treated with Rafoxanide or Closantel at the indicated concentrations or STOCK1S5 0699 (10 μM) for 30 min and then either left untreated or treated with hypotonic buffer for 30 min to activate WNK-SPAK/OSR1 signalling. The cells were then harvested, and the lysates were probed for phospho-NKCC1 T203, T207 and T212, total NKCC1, phospho-SPAKS373, total SPAK and GAPDH as a loading control. |
| Concentrations | 1, 3, 10, 20, 50 μM |
| Incubation time | 30 min |
| Animal Experiment | |
|---|---|
| Animal models | |
| Formulation | |
| Dosages | |
| Administration | |
| Molecular Weight | 626.01 |
| Formula | C19H11Cl2I2NO3 |
| CAS Number | 22662-39-1 |
| Solubility (25°C) | DMSO: ≥ 60 mg/mL |
| Storage |
Powder -20°C 3 years ; 4°C 2 years In solvent -80°C 6 months ; -20°C 1 month |
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