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STF-118804 is a highly specific NAMPT inhibitor.
| Cell Experiment | |
|---|---|
| Cell lines | Human cell lines (SEM, MV411, Nalm6, 697, Hal01, REH, SUP-B15 and KP-Y-RL) and primary patient leukemic samples |
| Preparation method | Human cell lines or lineage negative cord blood cells were seeded into 96-well plates (6?05 cells per milliliter). Adding compounds in increasing concentrations, and incubating cells at 37°C/5% CO2 for 72 hours. To detect viability, CellTiter-Blue reagent is added at 1:10 dilution, and plates are incubated for 4 hours at 37°C/5% CO2 prior to reading on a Flexstation II 384 or a Synergy H1 reader at an excitation of 555 nm and emission detection of 590 nm. Viability of cell measurement is also by CellTiter-Fluor. The cell-permeable fluorogenic peptide substrate GF-AFC reagent is added at the dilution of 1:2 , and plates are incubated for 30 min at 37°C/5% CO2 prior to reading on a Synergy H1 reader at an excitation of 380 nm and emission detection of 505 nm. Cord blood cells are enumerated on a hemocytometer, and cell viability is assessed with trypan blue exclusion dye. Using Prism software to calculate inhibitory concentration (IC50) . Primary patient samples are plated in 96-well plates and treated with increasing concentrations of STF-118804 for 48 hours at 37°C in 5% CO2. WST-1 reagent is added to the culture medium (1:10 dilution), and absorbance is measured at 450 nm using a Bio-Rad model 680 microplate reader. All assays are performed in triplicate. Using CalcuSyn version 2.0 software to calculate IC50 . |
| Concentrations | ~10 μM |
| Incubation time | 72 hours |
| Animal Experiment | |
|---|---|
| Animal models | Orthotopic xenograft model of ALL transplanted with MV411 cells |
| Formulation | 20% [w/v] [2-hydroxypropyl]-γ-cyclodextrin/5% [v/v] DMSO |
| Dosages | 25 mg/kg twice daily |
| Administration | s.c. |
| Molecular Weight | 461.53 |
| Formula | C25H23N3O4S |
| CAS Number | 894187-61-2 |
| Form | Solid |
| Solubility (25°C) | DMSO 50 mg/mL |
| Storage |
Powder -20°C 3 years ; 4°C 2 years In solvent -80°C 6 months ; -20°C 1 month |
[4] Szu-Chieh Mei, et al. NAD as a genotype-specific drug target
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