SRT 2104 (GSK2245840) is a selective and brain-permeable SIRT1 activator.

SRT2104 在 C2C12 细胞中降低 p65/RelA 的乙酰化水平。[1]

在雄性C57BL/6J小鼠中，SRT2104（100 mg/kg，口服）延长了标准饮食喂养小鼠的平均寿命和最大寿命，并提高了运动协调能力、骨密度、胰岛素敏感性，并降低了炎症。SRT2104的短期治疗能在萎缩实验模型中保持骨骼和肌肉质量。[1] 在雄性N171-82Q HD小鼠中，包含0.5% SRT2104的饮食能有效穿透血脑屏障，减缓脑萎缩，改善运动功能，并延长生存期。[2]

SIRT1 fluorescence polarization assay and HTS: In the SIRT1 FP assay, SIRT1 activity is monitored using a 20 amino acid peptide (Ac-Glu-Glu-Lys(biotin)-Gly-Gln-Ser-Thr-Ser-Ser-His-Ser-Lys(Ac)-Nle-Ser-Thr-Glu-Gly–Lys(MR121 or Tamra)-Glu-Glu-NH2 ) derived from the sequence of p53. The peptide is N-terminally linked to biotin and C-terminally modified with a fluorescent tag. The reaction for monitoring enzyme activity is a coupled enzyme assay where the first reaction is the deacetylation reaction catalyzed by SIRT1 and the second reaction is cleavage by trypsin at the newly exposed lysine residue. The reaction is stopped and streptavidin is added in order to accentuate the mass differences between substrate and product. The fluorescence polarization reaction conditions are as follows: 0.5 μM peptide substrate, 150 μM βNAD +, 0-10 nM SIRT1, 25 mM Tris-acetate pH 8, 137 mM Na-Ac, 2.7 mM K-Ac, 1 mM Mg-Ac, 0.05% Tween-20, 0.1% Pluronic F127, 10 mM CaCl 2 , 5 mM DTT, 0.025% BSA, and 0.15 mM nicotinamide. The reaction is incubated at 37°C and stopped by addition of nicotinamide, and trypsin is added to cleave the deacetylated substrate. This reaction is incubated at 37 ℃ in the presence of 1 μM streptavidin. Fluorescent polarization is determined at excitation (650 nm) and emission (680 nm) wavelengths.

Cells are cultured in low glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum and penicillin–streptomycin. Cells are treated with vehicle (0.1% DMSO) or 3?μM?SRT2104 for 24?h and then harvested for protein and Western blotting.(Only for Reference)