|Solubility (25°C) *||In vitro||DMSO||49 mg/mL (197.01 mM)|
|Ethanol||18 mg/mL (72.37 mM)|
|In vivo||5% DMSO+30% PEG 300+ddH2O||15mg/mL|
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
|Description||Selisistat (EX 527) is a potent and selective SIRT1 inhibitor with IC50 of 38 nM in a cell-free assay, exhibits >200-fold selectivity against SIRT2 and SIRT3. Phase 2.|
|In vitro||EX 527 exhibits potently inhibitory effect against SIRT1 deacetylase activity in a concentration-dependent manner with an IC50 of 38 nM, displays much lower activity against SIRT2 and SIRT3 with IC50 values of 19.6 μM and 48.7 μM, respectively. EX 527 does not inhibit SIRT4-7 and class I/II HDAC activity at concentrations up to 100 μM. EX-527 alone (1 μM) has no detectable effect on the acetylation of p53 lysine 382 in NCI-H460 cells. EX-527 significantly increases the amount of acetylated p53 in NCI-H460 cells, human mammary epithelial cells, U-2 OS and MCF-7 cells subjected to genotoxic agents such as Etoposide, Doxorubicin, Hydroxyurea, and Hydrogen peroxide, which is more effective than that caused by Nicotinamide (5 mM). But surprisingly EX 527 does not result in detectable effects on p53-controled gene expression, cell survival, or cell proliferation.  EX 527 causes a 90% increase in cell number of HCT116 cells after 7 days in the condition of 0.1% serum but not 10% serum, suggesting that SIRT1 is a significant regulator of cell proliferation during growth factor deprivation conditions.  EX 527 abrogates resveratrol effects on glucose responses, and prevents resveratrol-induced up-regulation of Glut2, glucokinase, Pdx-1, and Tfam in INS-1E Cells, due to the opposite effect of EX 527 and resveratrol on SIRT1 deacetylase activity. |
|In vivo||Administration of EX 527 (~10 μg) to rats increases hypothalamic acetyl-p53 levels by inhibiting hypothalamic SIRT1 activity. Co-administration of EX 527 with ghrelin markedly blunts the orexigenic action of ghrelin by decreasing the pAMPK levels, increasing the ACC levels, and abolishing the higher expression of the transcription factors FoxO1, pCREB, and Bsx and the neuropeptides NPY and AgRP in the hypothalamic arcuate nucleus. |
|Features||Greater potency, specificity, stability, and lower toxicity than other inhibitors of SIRT1 catalytic activity identified to date.|
|Inhibition of GST-SIRT1 deacetylase activity||293T cells are transiently transfected with GST-tagged human SIRT1 in the pDEST27 Gateway vector using FuGENE-6. After 48 hours, the cells are lysed with 50 mM Tris, pH 8.0, 120 mM NaCl, 1 mM EDTA, and 0.5% Nonidet P-40, supplemented with Complete Mini protease inhibitor cocktail tablets. GST-SIRT1 is purified from lysates using glutathione-Sepharose beads and washed extensively in the above buffer. The deacetylation assay is performed with approximately 30 ng of GST-SIRT1 in the presence of EX 527 (48 pM to 100 μM). Deacetylation is measured using the Fluor de Lys kit using a fluorogenic peptide encompassing residues 379 to 382 of p53, acetylated on lysine 382. The acetylated lysine residue is coupled to an aminomethylcoumarin moiety. The peptide is deacetylated by SIRT1, followed by the addition of a proteolytic developer that releases the fluorescent aminomethylcoumarin. Briefly, enzyme preparations are incubated with 170 μM NAD+ and 100 μM p53 fluorogenic peptide for 45 minutes at 37 °C followed by incubation in developer for 15 minutes at 37 °C. Fluorescence is measured by excitation at 360 nm and emission at 460 nm and enzymatic activity is expressed in relative fluorescence units.|
|Cell lines||NCI-H460, MCF-7, U-2 OS and HMEC|
|Concentrations||Dissolved in DMSO, final concentration 1 μM|
|Incubation Time||48 or 72 hours|
|Method||For viability assays, cells are treated with EX 527 for 48 hours. Cell viability is then determined using the Cell Titer-Glo luminescent assay, which measures total ATP level as an index of cell number. Luminescence is measured on a Luminoskan Ascent. For the proliferation assay, 0.5 μCi/mL of [14C]thymidine is added to the medium immediately after EX 527. Plates are counted at 48 hours (HMEC) or 72 hours (NCI-H460, MCF-7, and U-2 OS cells) in a Microbeta liquid scintillation counter. Thymidine incorporated by the cells is detected by proximity to the scintillant in the base of the Cytostar-T tissue culture plate.|
|Animal Models||Male Sprague-Dawley rats|
|Formulation||Dissolved in DMSO in a total volume of 5 μL|
Data from [Data independently produced by J Pineal Res, 2014, 57(2), 228-38]
The eect of Melatonin, luzindole, and EX527 treatment on apoptotic index, infarct size, lactate dehydrogenase (LDH) release, and CK release in IR-injured hearts. Representative photomicrographs of in situ detection of apoptotic myocytes by TUNEL staining. Green ﬂuorescence shows TUNEL-positive nuclei; blue ﬂuorescence shows nuclei of total cardiomyocytes; original magniﬁcation 9x400. Sacle bar: 100 um.
Data from [Cancer Res, 2014, 74(1), 298-308]
Cells were treated with CP, DOXO, SRT1720 (100 nM), EX527 (100 nM), A769662 (10 μM) and Compound C (1 μM) under normoxic or hypoxic conditions for 48 hours, and then their viabilities were measured by MTT.
SIRT1 prevents genotoxic stress-induced p53 activation in acute myeloid leukemia. [Girdwood SC, et al. Blood, 2014, 124(1):121-33]PubMed: 24855208
Melatonin receptor-mediated protection against myocardial ischemia/reperfusion injury: role of SIRT1 [Yu L et al. J Pineal Res, 2014, 57(2):228-38]PubMed: 25052362
SIRT1 and AMPK mediate hypoxia-induced resistance of non-small cell lung cancers to cisplatin and doxorubicin. [Shin DH, et al. Cancer Res, 2014, 74(1):298-308]PubMed: 24240701
G-quadruplex-based fluorometric biosensor for label-free and homogenous detection of protein acetylation-related enzymes activities. [Wang H, et al. Biosens Bioelectron, 2017, 91:400-407]PubMed: 28063389
HDAC4 mediates IFN-γ induced disruption of energy expenditure-related gene expression by repressing SIRT1 transcription in skeletal muscle cells [Fang M, et al. Biochim Biophys Acta, 2016, 1859(2):294-305]PubMed: 26619800
60-kDa Tat-interactive Protein (TIP60) Positively Regulates Th-inducing POK (ThPOK)-mediated Repression of Eomesodermin in Human CD4+ T Cells. [Li Y, et al. J Biol Chem, 2013, 288(22):15537-46]PubMed: 23609452
Histone deacetylase III as a potential therapeutic target for the treatment of lethal sepsis [Zhao T,et al. J Trauma Acute Care Surg, 2014, 77(6):913-9]
Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKKβ-AMPK-SIRT1 Signaling Pathway [Liu X, et al. Mediators Inflamm, 2016, 10.1155/2016/6152713]PubMed: 27313401
ZLN005 protects cardiomyocytes against high glucose-induced cytotoxicity by promoting SIRT1 expression and autophagy. [Li W, et al. Exp Cell Res, 2016, 345(1):25-36]PubMed: 27208585
Effect of the BRCA1-SIRT1-EGFR axis on cisplatin sensitivity in ovarian cancer [Li D, et al. Am J Transl Res, 2016, 8(3):1601-8]PubMed: 27186285
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