Thiamet-G - Autophagy Activator - 生命科學(xué)試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEThiamet GCat. No.: HY-12588CAS No.: 1009816-48-1分式: CHNOS分量: 248.3作靶點(diǎn): Autophagy作通路: Autophagy儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) H2O : 50 mg/mL (201.37 mM)DMSO : 45 mg/mL (181.23 mM)*

2、 means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 4.0274 mL 20.1369 mL 40.2739 mL5 mM 0.8055 mL 4.0274 mL 8.0548 mL10 mM 0.4027 mL 2.0137 mL 4.0274 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 Thiamet G 為有效，選擇性的 O-GlcNAcase 抑制劑，其作從修飾的蛋質(zhì)

3、中去除O-GlcNAc，抑制 OGA的 Ki 值為 20 nM。IC50 & Target Ki: 20 nM (Human OGA) 11/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE體外研究 Thiamet G (1 M) induces a clear increase in the accumulation of O-GlcNAcylated proteins of ATDC5 cells.O-GlcNAc accumulation induced by Thiamet G also evokes a clear increase in

4、 the activity of these MMPs.Thiamet G (1 M) induces the phosphorylation of JNK, ERK, and p38 but not phosphorylation of Akt 2.Thiamet G (0.1-10 M) does not significantly affect the cell viability. Thiamet G decreases phosphorylation oftau and alters the microtubule dynamics 3.體內(nèi)研究 Thiamet G (500 mg/

5、kg/d) increases global and tau O-GlcNAc and reduces neurodegeneration. Thiamet G-treated group has 1.4-fold more motor neurons and hinders tau-driven neurodegeneration within thistransgenic model. Thiamet G treatment therefore has no detectable effect on mice lacking the P301Ltransgene, indicating t

6、hat prevention of neurodegeneration and weight loss is mediated by Thiamet Gtreatment only in the context of the P301L transgene. In Thiamet G-treated mice, the O-GlcNAc increases inthe brain and spinal cord tissues 1. Thiamet G (20 mg/kg, i.p.) increases O-GlcNAc levels in brain, liver, andknee of

7、the C57BL/6 mice in a dose-dependent manner 2.PROTOCOLKinase Assay 3 All enzymatic assays are performed in triplicate at 37C using 4-methylumbelliferyl N-acetyl-d-glucosaminide dehydrate as substrate. 1 nM of purified OGA is incubated with the compounds for 5 min, andthen 0.2 mM of the substrate is

8、added. The liberation of 4-methylumbellifery is monitored by kinetic readingat excitation/emission 355/460 nm using a Tecan M200 plate in a mode of 60 s/cycle and 15 cycles in total.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 3 Jurkat cel

9、ls are seeded at 6000 cells/well in a 96-well plate, and 12 h later, cells are treated with compoundsfor the indicated time. Cell viability is determined by XTT assay.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal For the Thiamet G dose dependen

10、ce study, six 23-day-old male C57BL/6 mice receive single intraperitonealAdministration 2 injections of either 0, 10, 20, 100, 200, or 500 mg/kg of Thiamet G dissolved in phosphate-buffered saline(PBS) and then are euthanized 8 h later to evaluate the O-GlcNAc levels in different tissues (brain, liv

11、er,muscle, and knee). The time of sacrifice is chosen on the basis of previously published data on Thiamet G inrodents, which demonstrates that the peak level of O-GlcNAc proteins following administration of the drug isachieved after 8-10 h. Tissues are collected immediately after sacrifice, flash-f

12、rozen in liquid nitrogen, andstored at 80C until required for use.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Yuzwa SA, et al. Increasing O-GlcNAc slows neurodegeneration and stabilizes tau against aggregation. Nat Chem Biol. 2012 Feb26

13、;8(4):393-9.2. Andrs-Bergs J, et al. The increase in O-linked N-acetylglucosamine protein modification stimulates chondrogenic differentiation bothin vitro and in vivo. J Biol Chem. 2012 Sep 28;287(40):33615-28.3. Ding N, et al. Thiamet-G-mediated inhibition of O-GlcNAcase sensitizes human leukemia cells to microtubule-stabilizing agent paclitaxel.Biochem Biophys Res Commun. 2014 Oct 24;453(3):392-7.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChem