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M344 shows a potent inhibition activity against histone deacetylase with IC50 of 100 nM. M344 produces a more significant effect on cell proliferation than on cell differentiation in MEL DS19 cells. M344 shows cell toxicity at concentrations above 10 μM, while a maximum of only 20% of the surviving cell population are induced to differentiate. In vitro, M344 produces the significant anti-proliferative effects on the endometrial cancer cell line Ishikawa and the ovarian cancer cell line SK-OV-3 with EC50 of 2.3 μM and 5.1 μM, respectively. While the normal human endometrial epithelial cells shows little sensitivity to M344. In addition, M344 also decreases proportion of cells in the S-phase, increases proportion in the G0/G1 phases of the cell cycle, induces apoptosis and decreases the transmembrane potential of mitochondria. M344 show the anti-proliferative activities against embryonal nervous system tumor cells including medulloblastoma cells (D341 Med, Daoy) and neuroblastoma cells (CH-LA 90,SHSY-5Y ) with GI50 of 0.65 μM, 0.63 μM, 0.63 μM and 0.67 μM, respectively.M344 (5 μM ) in combination with cisplatin (10 μg/mL) results increased induction of ATF3 and shows enhanced cytotoxic effects than cisplatin alone in a panel of human derived cancer cell lines, such as MCF-7, PC3, SK-OV3, and A549.
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Source | Cell Death Dis (2018). Figure 1. M344 |
| Method | cell viability assay | |
| Cell Lines | T-ALL cells | |
| Concentrations | 1 μM | |
| Incubation Time | 24 and 48 h | |
| Results | Compared with the single treatment (Fig. 1a–c), the combination of vorinostat and QC significantly decreased cell viability. |
| Cell Experiment | |
|---|---|
| Cell lines | MEL DS19 cells |
| Preparation method | Maintaining MEL DS19 cells (murine erythroleukemia cells) in D-MEM containing 100 units/mL penicillin G sodium and 100 μg/mL streptomycin sulfate supplemented with 10% fetal bovine serum at 37 °C in a 5% CO2 atmosphere. To test M344 for potential to induce cell differentiation, log-phase cells with a population doubling time of 11−13 hours are used. Preparing Serial dilutions of M344 in 24-well plates using 1 mL of D-MEM/well. If M344 are dissolved in DMSO, control wells contains the same amount of solvent (generally 2 μL/mL of medium). Subsequently, adding the cell suspension to the wells. After 72 hours the experiment is evaluated. Using a Casy 1 TTC flow cytometer to counte cell numbers. With the solvent control , expressing the proliferation of treated cells as percent proliferation in comparison. Differentiated MEL cells accumulate hemoglobin. Therefore, the induction of cell differentiation is determined by benzidine staining according to the literature. To 100 μL of cell suspension is added 10 μL of a 0.4% solution of benzidine in 12% acetic acid which contains 2% H2O2. Within 5 minutes hemoglobin-containing cells stains blue.Under the microscope in a hemocytometer ,counting Benzidine-positive and -negative cells , and calculating the percentage of positive cells . Testing M344 first at 10 μM and 50 μM final concentration. A range of concentrations is chosen for a dose−response analysis according to activity/toxicity profile,. |
| Concentrations | 0 to 50 μM |
| Incubation time | 72 hours |
| Animal Experiment | |
|---|---|
| Animal models | |
| Formulation | |
| Dosages | |
| Administration | |
| Molecular Weight | 307.39 |
| Formula | C16H25N3O3 |
| CAS Number | 251456-60-7 |
| Solubility (25°C) | DMSO 55 mg/mL |
| Storage |
Powder -20°C 3 years ; 4°C 2 years In solvent -80°C 6 months ; -20°C 1 month |
[4] Angie Yeung, et al. HDAC inhibitor M344 suppresses MCF-7 breast cancer cell proliferation
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