CY-09 - NOD-like Receptor (NLR) 抑制劑 - 生命科學(xué)試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemECY-09Cat. No.: HY-103666CAS No.: 1073612-91-5分式: CHFNOS分量: 423.43作靶點(diǎn): NOD-like Receptor (NLR)作通路: Immunology/Inflammation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) DMSO : 150 mg/mL (354.25 mM

2、)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 2.3617 mL 11.8083 mL 23.6167 mL5 mM 0.4723 mL 2.3617 mL 4.7233 mL10 mM 0.2362 mL 1.1808 mL 2.3617 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(shí)驗(yàn) 請根據(jù)您的實(shí)驗(yàn)動物和給藥式選擇適當(dāng)?shù)娜芙獍福渲魄罢埾扰渲瞥吻宓膬湟?，再依次添加助溶?為保證實(shí)驗(yàn)結(jié)果的可靠性，體內(nèi)

3、實(shí)驗(yàn)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)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 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.90 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.90 mM); Clear solution1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEBI

4、OLOGICAL ACTIVITY物活性 CY-09是NLRP3 的抑制劑。IC50 & Target NLRP3 1體外研究 CY-09 exhibits a dose-dependent inhibitory effect on monosodium urate (MSU), nigericin, ATP-inducedcaspase-1 activation and IL-1 secretion at the doses of 1 to10 M in LPS-primed bone marrow-derivedmacrophages (BMDMs). Cytosolic LPS-indu

5、ced noncanonical NLRP3 activation in BMDMs can also beblocked by CY-09 treatment. CY-09 specifically inhibits NLRP3 inflammasome activation and has no effecton LPS-induced priming effects. CY-09 treatment remarkably suppresses nigericin-induced ASColigomerization. It is found that CY-09 treatment in

6、hibits the interaction of Flag-NLRP3 and mCherry-NLRP3in HEK-293T cells, suggesting that CY-09 blocks NLRP3 oligomerization 1.體內(nèi)研究 CY-09 treatment in vivo efficiently suppresses monosodium urate (MSU) injection-induced IL-1 productionand neutrophil influx, suggesting that CY-09 can block MSU-induced

7、 NLRP3 inflammasome activation invivo. CY-09 treatment also increases the survival of NLRP3 mutant mice up to days 30 to 48 even aftertreatment is stopped at day 25. The caspase-1 cleavage observed in adipose tissue of high-fat diet (HFD)-treated mice is also suppressed by CY-09 1.PROTOCOLKinase Ass

8、ay 1 For ATPase activity assay, purified recombinant human proteins are incubated at 37C with indicatedconcentrations of CY-09 for 15 min in the reaction buffer. ATP (25 m) is then added, and the mixture isfurther incubated at 37C for another 40 min. The amount of ATP converted into adenosine diphos

9、phate(ADP) is determined by luminescent ADP detection with ADP-Glo Kinase Assay kit according to themanufacturers protocol. The results are expressed as percentage of residual enzyme activity to the vehicle-treated enzyme. For ATP binding assay, purified NLRP3 proteins are incubated with ATP binding

10、 agarose for1 h and then different concentrations of CY-09 are added and incubated for 2 h with motion at 4C. Beadsare washed and boiled in loading buffer. Samples are subjected to immunoblotting analysis 1.MCE has not independently confirmed the accuracy of these methods. They are for reference onl

11、y.Cell Assay 1 To induce NLRP3 inflammasome activation, 5105/mL BMDMs and 6106/mL PBMCs are plated in 12-wellplates. The following morning, the medium is replaced, and cells are stimulated with 50 ng/mL LPS or 400ng/mL Pam3CSK4 (for noncanonical inflammasome activation) for 3 h. After that, CY-09 or

12、 other inhibitorsare added into the culture for another 30 min, and then the cells are stimulated for 4 h with monosodiumurate (MSU) (150 g/mL), Salmonella typhimurium (multiplicity of infection) or for 30 min with ATP (2.5 mM)or nigericin (10 M). Cells are transfected with poly(dA:dT) (0.5 g/mL) fo

13、r 4 h or LPS (500 ng/mL) overnight.Cell extracts and precipitated supernatants are analyzed by immunoblot 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal WT or Nlrp3/ mice at the age of 6 wk, with similar plasma glucose levels and body weights

14、are randomizedAdministration 1 into different groups. For generation of high-fat diet (HFD)-induced diabetic mice, mice are fed with HFD for14 wk. The diabetic mice are treated with CY-09 (i.p.) at a dose of 2.5 mg/kg once a day for 6 wk. The mice2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEare

15、maintained with HFD when used for CY-09 treatment and the subsequent experiments 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Jiang H, et al. Identification of a selective and direct NLRP3 inhibitor to treat inflammatory disorders. J Exp Med. 201