| 名稱(chēng) | MCC950 sodium |
| 描述 | MCC950 sodium (CP-456773 sodium) is a potent and selective inhibitor of the inflammatory vesicle NLRP3 (IC50=7.5 nM in BMDMs; IC50=8.1 nM in HMDMs). MCC950 sodium has no effect on other inflammatory vesicles such as AIM2, NLRC4 or NLRP1. |
| 細(xì)胞實(shí)驗(yàn) | MCC950 is dissolved in DMSO and stored, and then diluted with appropriate media before use[1]. BMDM are seeded at 5×105/mL or 1×106/mL, HMDM at 5×105/mL and PBMC at 2×106/mL or 5×106/mL in 96 well plates. The following day the overnight medium is replaced and cells are stimulated with 10 ng/mL LPS from Escherichia coli serotype EH100 (ra) TLRgrad for 3 h. Medium is removed and replaced with serum free medium (SFM) containing DMSO (1:1,000), MCC950 (0.001-10 μM), glyburide (200 μM), Parthenolide (10 μM) or Bayer cysteinyl leukotriene receptor antagonist 1-(5-carboxy-2{3-[4-(3-cyclohexylpropoxy)phenyl]propoxy}benzoyl)piperidine-4-carboxylic acid (40 μM) for 30 min. Cells are then stimulated with inflammasome activators: 5 mM adenosine 5'-triphosphate disodium salt hydrate (ATP) (1 h), 1 μg/mL Poly(deoxyadenylic-thymidylic) acid sodium salt (Poly dA:dT) transfected with Lipofectamine 200 (3-4 h), 200 μg/mL MSU (overnight) and 10 μM nigericin (1 h) or S. typhimurium UK-1 strain. Cells are also stimulated with 25 μg/mL Polyadenylic-polyuridylic acid (4 h). For non-canonical inflammasome activation cells are primed with 100 ng/mL Pam3CSK4 for 4 h, medium is removed and replaced with SFM containing DMSO or MCC950 and 2 μg/mL LPS is transfected using 0.25% FuGENE for 16 h. Supernatants are removed and analysed using ELISA kits. LDH release is measured using the CytoTox96 non-radioactive cytotoxicity assay[1]. |
| 激酶實(shí)驗(yàn) | kinase activity assays: All assays are carried out in 384-well white microtiter plates. Compounds are 4-fold serially diluted in 8 steps, starting from 10 μM. The reaction mixture consisted of 25 μL assay buffer (50 mM HEPES pH 7.5, 10 mM MgCl2, 5 mM MnCl2, 1 mM DTT, 0.1 mM Na3VO4, 5 mM β-glycerol phosphate). For FLT3 assays, the reaction contains 2.0 μg/mL FLT3 enzyme, 5 μM of poly(Glu,Tyr) substrate and 4 μM of ATP. For JAK1 assays, the reaction contains 2.5 μg/mL of JAK1 enzyme, 10 μM of poly(Glu,Ala,Tyr) substrate and 1.0 μM of ATP. For JAK2 assays, the reaction contained 0.35 μg/mL of JAK2 enzyme, 10 μM of poly (Glu,Ala,Tyr) substrate and 0.15 μM of ATP. For JAK3 assays, the reaction contained 3.5 μg/mL of JAK3 enzyme, 10 μM of poly (Glu,Ala,Tyr) substrate and 6.0 μM of ATP. For TYK2 assays, the reaction contained 2.5 μg/mL of TYK2 enzyme, 10 μM of poly (Glu,Ala,Tyr) substrate and 0.15 μM of ATP. The reaction is incubated at room temperature for 2 h prior to addition of 13 μL PKLight? detection reagent. After 10 min incubation luminescent signals are read on a multi-label plate reader. |
| 動(dòng)物實(shí)驗(yàn) | C57BL/6 mice were injected intraperitoneally with 50 mg/kg MCC950. |
| 體外活性 | 方法：小鼠骨髓衍生的巨噬細(xì)胞 BMDM 用 LPS (10 ng/mL) 刺激 3 h，用 MCC950 sodium (1-1000 nM) 刺激 30 min，再用 ATP (5 mM) 處理 1 h，使用 ELISA 方法檢測(cè) IL-1β 和 TNF-α 水平。
結(jié)果：MCC950 處理細(xì)胞可劑量依賴(lài)性地抑制 BMDM 中 IL-1β 的釋放。LPS 依賴(lài)性 TNF-α 分泌未被 MCC950 損害。[1]
方法：小鼠巨噬細(xì)胞、人冠狀動(dòng)脈內(nèi)皮細(xì)胞和平滑肌細(xì)胞用 MCC950 sodium (0.02-20 μM) 處理 3 天，使用 Alamar Blue assay 檢測(cè)細(xì)胞活力。
結(jié)果：MCC950 sodium 對(duì)三種細(xì)胞無(wú)毒性作用。[2] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗 NLRP3 活性，將 MCC950 sodium (20 mg/kg) 腹腔注射給人 CAPS 疾病 MWS 的小鼠模型，每天一次，持續(xù)四周。
結(jié)果：MCC950 拯救 CAPS 小鼠模型并抑制人 MWS 細(xì)胞中的 NLRP3。[1]
方法：為研究對(duì)體內(nèi)實(shí)驗(yàn)性脊髓損傷模型和體外神經(jīng)元損傷的藥理作用，將 MCC950 sodium (10-50 mg/kg) 腹腔注射給脊髓損傷 (SCI) 的 C57BL/6 小鼠。
結(jié)果：MCC950 改善了 SCI 小鼠的握力、后肢運(yùn)動(dòng)、脊髓水腫和病理?yè)p傷。它通過(guò)阻斷 NLRP3 炎癥小體組裝以及促炎細(xì)胞因子 TNF-α、IL-1β 和 IL-18 的釋放來(lái)發(fā)揮這種作用。[3] |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | H2O : 42.7 mg/mL (100 mM)
DMSO : 55 mg/mL (128.97 mM)
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| 關(guān)鍵字 | MCC 950 | MCC-950 sodium | inhibit | CP456773 | Inhibitor | NOD-like Receptor (NLR) | CRID3 | CP 456773 | MCC-950 | CRID 3 | CRID3 sodium | MCC950 sodium | CRID-3 |
| 相關(guān)產(chǎn)品 | Bergenin | JC2-11 | Selnoflast | NLRP3-IN-2 | Imperatorin | INF39 | NLRP3-IN-10 | Dapansutrile | Stavudine | NOD-IN-1 | Muscone | Troxerutin |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 細(xì)胞焦亡化合物庫(kù) | 經(jīng)典已知活性庫(kù) | 已知活性化合物庫(kù) | 高選擇性抑制劑庫(kù) | 抗衰老化合物庫(kù) | HIF-1化合物庫(kù) | 非甾體類(lèi)抗炎化合物庫(kù) | 抗卵巢癌化合物庫(kù) | 抗抑郁癥化合物庫(kù) |