P-22077 _Deubiquitinase 抑制劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEP 22077Cat. No.: HY-13865CAS No.: 1247819-59-5分式: CHFNOS分量: 315.32作靶點: Deubiquitinase作通路: Cell Cycle/DNA Damage儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 DMSO : 50 mg/mL (158.57 mM; Need ultr

2、asonic)Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 3.1714 mL 15.8569 mL 31.7138 mL5 mM 0.6343 mL 3.1714 mL 6.3428 mL10 mM 0.3171 mL 1.5857 mL 3.1714 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。體內(nèi)實驗 請根據(jù)您的實驗動物和給藥式選擇適當(dāng)?shù)娜芙獍?，配制前請先配制澄清的儲備液，再依次添加助溶?為保證實驗結(jié)果的可靠性，體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄清的儲備液可以根據(jù)儲存條件，適當(dāng)

3、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. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (7.93 mM); Clear solutionBIOLOGICAL ACTIVITY物活性P 22077 種泛素蛋特異性蛋酶 (USP7) 抑制劑，EC50 值為 8.01 M，同時可抑制 USP47，EC50 值為1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE8.74 M。IC50 & Target EC50: 8.01 M (USP7), 8.74 M (USP4

4、7) 1體外研究 P 22077 is an inhibitor of USP7 and DUB USP47, with EC50s of 8.01 M and 8.74 M, respectively. P 22077(15-45 M) inhibits a much smaller subset of DUBs. P 22077 (25 M) causes DUBs inhibition in HEK293Tcells 1. P 22077 (0-20 M) greatly reduces the cell viability of Neuroblastoma (NB) cells inc

5、luding IMR-32,NGP, CHLA-255, and SH-SY5Y cells but without NB-19 and SK-N-AS cells. P 22077 (10 M) increases p53activity and induces apoptosis in p53 wild-type and HDM2-expressing NB cells. P 22077 (5 M) enhancesthe cytotoxic effect of Dox and VP-16 on NB cells, and enhances Dox- and VP-16-induced p

6、53-mediatedapoptosis 2.體內(nèi)研究 P 22077 (15mg/kg, i.p. 21 days) shows potent antitumor activities in an xenograft mouse model bearing IMR-32-derived tumors; P 22077 also exhibits antitumor effects after treatment at 10mg/kg for 14 days in micebearing SH-SY5Y-derived tumors, and at 20 mg/kg for 12 days i

7、n mice bearing NGP-derived tumors 2.PROTOCOLKinase Assay 1 Recombinant full length USP7, USP2 core, USP5, JOSD2, DEN1, PLpro core, and SENP2 catalytic core aregenerated. Amino terminal His6 tagged USP4, USP8, USP28, UCH-L1, UCH-L3, UCH-L5, and MMP13 areexpressed in Escherichia coli. N-terminal His6

8、tagged USP15, USP20, and USP47 are expressed in Sf9cells. All the recombinant proteins are purified by chromatography. Amino terminal tagged His6 Ub-PLA2(Ub-CHOP), SUMO3-PLA2 (SUMO3-CHOP), ISG15-PLA2 (ISG15-CHOP), NEDD8-PLA2 (NEDD8-CHOP),Ub-EKL (Ub-CHOP2), and free catalytically active PLA2 are prep

9、ared 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 Cell viability assays are assessed using the Cell Counting Kit-8 (CCK-8, WST-82-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt). Cells

10、 are seeded in 96-well flat-bottomed plates at the density of 1 104 per well. After 24h of incubation at 37C, increasing concentrationsof P 22077, Dox, VP-16, or their combinations are added to the wells. Twenty-four hours later, 10L of CCK-8 is added into each well and after 1h of incubation, the a

11、bsorbance is measure at 450nm using themicroplate reader. Each experiment is performed in replicates of six. Background reading of media only isused to normalize the results 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal The orthotopic Neurobl

12、astoma (NB) mouse model is used in the assay. Briefly, 1.5 106 human IMR-32,Administration 2 SH-SY5Y, or NGP cells with luciferase expression are surgically injected into the left renal capsule of 5-week-old female NCR nude mice. IMR-32, SH-SY5Y, and NGP-derived xenografts are allowed to grow for2-3

13、 weeks before randomizing the mice into a control group and a P 22077 treatment group. Each groupconsists of three or six mice. Animals are treated with DMSO or P 22077 by intraperitoneal (i.p.) injectionevery day for 12, 14, or 21 days. At the end of the experiments, all mice are killed. Tumors and

14、 the right sidecontrol kidneys are resected, weighed, and photographed 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE戶使本產(chǎn)品發(fā)表的科研獻 Nat Chem Biol. 2017 Dec;13(12):1207-1215. Haematologica. 2019 Mar 14.Se

15、e more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Altun M, et al. Activity-based chemical proteomics accelerates inhibitor development for deubiquitylating enzymes. Chem Biol. 2011 Nov23;18(11):1401-12.2. Fan YH, et al. USP7 inhibitor P22077 inhibits neuroblastoma growth via inducing p53-mediated apoptosis. Cell Dea