Tubastatin-A-Hydrochloride-Tubastatin-A-HCl-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemETubastatin A HydrochlorideCat. No.: HY-13271CAS No.: 1310693-92-5Synonyms: Tubastatin A HCl; TSA HCl分式: CHClNO分量: 371.86作靶點(diǎn): HDAC; Autophagy作通路: Cell Cycle/DNA Damage; Epigenetics; Autophagy儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn s

2、olvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 10.8 mg/mL (29.04 mM; Need ultrasonic and warming)Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 2.6892 mL 13.4459 mL 26.8918 mL5 mM 0.5378 mL 2.6892 mL 5.3784 mL10 mM 0.2689 mL 1.3446 mL 2.6892 mL請(qǐng)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(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存式和期限。體內(nèi)實(shí)驗(yàn)請(qǐng)根

3、據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍福渲魄罢?qǐng)先配制澄清的儲(chǔ)備液，再依次添加助溶劑(為保證實(shí)驗(yàn)結(jié)果的可靠性，體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄清的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；以下溶劑前的百分 指該溶劑在您配制終溶液中的體積占)：1. 請(qǐng)依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.08 mg/mL (5.59 mM); Clear solution2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.08 mg/mL (5.

4、59 mM); Clear solution3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oil1/4 Master of Small Molecules 您邊的抑制劑師www.MedChemESolubility: 2.08 mg/mL (5.59 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 Tubastatin A Hydrochloride種有效的，選擇性的 HDAC6 抑制劑，IC50 值為 15 nM，對(duì)其選擇性是HDAC8 外的其他亞型的 1000 多倍。IC50 & Target HDAC6 HDAC8 HDAC115 n

5、M (IC50) 854 nM (IC50) 16400 nM (IC50)體外研究 Tubastatin A is substantially selective for all 11 HDAC isoforms and maintains over 1000-fold selectivityagainst all isoforms excluding HDAC8, where it has approximately 57-fold selectivity. In homocysteic acid(HCA) induced neurodegeneration assays, Tubasta

6、tin A displays dose-dependent protection against HCA-induced neuronal cell death starting at 5 M with near complete protection at 10 M 1. At 100 ng/mLTubastatin A increases Foxp3+ T-regulatory cells (Tregs) suppression of T cell proliferation in vitro2. Tubastatin A treatment in CC12 cells would lea

7、d to myotube formation impairment when alpha-tubulin ishyperacetylated early in the myogenic process; however, myotube elongation occurs when alpha-tubulin ishyeperacetylated in myotubes 3. A recent study indicates that Tubastatin A treatment increases cellelasticity as revealed by atomic force micr

8、oscopy (AFM) tests without exerting drastic changes to the actinmicrofilament or microtubule networks in mouse ovarian cancer cell lines, MOSE-E and MOSE-L 4.體內(nèi)研究 Daily treatment of Tubastatin A at 0.5 mg/kg inhibits HDAC6 to promote Tregs suppressive activity in mousemodels of inflammation and auto

9、immunity, including multiple forms of experimental colitis and fully majorhistocompatibility complex (MHC)-incompatible cardiac allograft rejection 2.PROTOCOLKinase Assay 1 Enzyme inhibition assays are performed using the Reaction Biology HDAC Spectrum platform. The HDAC1,2, 4, 5, 6, 7, 8, 9, 10, an

10、d 11 assays use isolated recombinant human protein; HDAC3/NcoR2 complex isused for the HDAC3 assay. Substrate for HDAC1, 2, 3, 6, 10, and 11 assays is a fluorogenic peptide fromp53 residues 379-382 (RHKKAc); substrate for HDAC8 is fluorogenic diacyl peptide based on residues 379-382 of p53 (RHKAcKAc

11、). Acetyl-Lys (trifluoroacetyl)-AMC substrate is used for HDAC4, 5, 7, and 9 assays.Tubastatin A is dissolved in DMSO and tested in 10-dose IC50 mode with 3-fold serial dilution starting at 30 M. Control Compound Trichostatin A (TSA) is tested in a 10-dose IC50 with 3-fold serial dilution starting a

12、t 5M. IC50 values are extracted by curve-fitting the dose/response slopes.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Primary cortical neuron cultures are obtained from the cerebral cortex of fetal Sprague-Dawley rats(embryonic day 17).

13、 All experiments are initiated 24 hours after plating. Under these conditions, the cells arenot susceptible to glutamate-mediated excitotoxicity. For cytotoxicity studies, cells are rinsed with warm PBSand then placed in minimum essential medium containing 5.5 g/L glucose, 10% fetal calf serum, 2 mM

14、 L-glutamine, and 100 M cystine. Oxidative stress is induced by the addition of the glutamate analogue2/4 Master of Small Molecules 您邊的抑制劑師www.MedChemEhomocysteate (HCA; 5 mM) to the media. HCA is diluted from 100-fold concentrated solutions that areadjusted to pH 7.5. In combination with HCA, neuro

15、ns are treated with Tubastatin A at the indicatedconcentrations. Viability is assessed after 24 hours by MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal The effects o

16、f HDAC6 targeting in dextran sodium sulfate (DSS) and adoptive transfer models of colitis areAdministration 2 evaluated, using 10 mice per group. Freshly prepared 4% (wt/vol) DSS is added daily for 5 days to the pH-balanced tap water of WT B6 mice. Mice are treated daily for 7 days with tubacin or n

17、iltubacin (0.5 mg/kg ofbody weight/day, i.p.), and colitis is assessed by daily monitoring of body weight, stool consistency, and fecalblood. Stool consistency is scored as 0 (hard), 2 (soft), or 4 (diarrhea), and fecal blood (Hemoccult) is scoredas 0 (absent), 2 (occult), or 4 (gross). To assess pr

18、evention of colitis in a T cell-dependent model, CD4+CD45RBhi T cells (1106) isolated from WT mice using magnetic beads (95% cell purity, flow cytometry)are injected i.p. into B6/Rag1/ mice plus CD4+ CD25+ Tregs (1.25105) isolated using magnetic beadsfrom HDAC6/ or WT mice (90% Treg purity, flow cyt

19、ometry) and mice are monitored biweekly for clinicalevidence of colitis. To assess therapy of established T cell-dependent colitis, B6/Rag1/ mice are injectedi.p. with CD4+ CD45RBhi cells (1106). Once colitis has developed, mice also receive CD4+ CD25+ Tregs(5105 cells) isolated as described above f

20、rom HDAC6/ or WT mice or treatment with HDAC6i (tubastatinA) or HSP90i (17-AAG). Mice are monitored for continued weight loss and stool consistency. At the cessationof the study, paraffin sections of colons stained with Alcian Blue or hematoxylin and eosin are gradedhistologically or evaluated by im

21、munoperoxidase staining for Foxp3+ Treg infiltration.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) BMC Biol. 2018 Oct 18;16(1):116. Mol Cancer Res. 2014 May;12(5):681-93. Front Pharmacol. 2018 Feb 1;9:34. Patent. US20180263995A1. Medical S

22、cience, University of Toronto. 2017 Nov.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Kyle V. Butler et al. Rational Design and Simple Chemistry Yield a Superior, Neuroprotective HDAC6 Inhibitor, Tubastatin A J. Am.Chem. Soc., 2010, 132 (31), pp 10842-108462. de Zoeten EF, et al. Histone deacetylase 6 and heat shock protein 90 control the functions of Foxp3(+) T-regulatory cells. Mol Cell Biol.2