MCC950 (CP-456773)
(Synonyms: CP-456773; CRID3) 目录号 : GC31644
An inhibitor of NLRP3 inflammasome activation
Cas No.:210826-40-7
Sample solution is provided at 25 µL, 10mM.
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Related Biological Data
DBP exposure activated LX-2 cells via p38MAPK/NF-κB/NLRP3-mediated pyroptosis. (B, D) The images of α-SMA, TGF-β, and Col-Ⅰ protein expression in LX-2 cells. (C, E) The quantitative analysis of α-SMA, TGF-β,andCol-Ⅰ protein expression in LX-2 cells.
Before DBP stimulation, the p38MAPK (SB203580, 5 µM) , NF-κB (PDTC, 10 µM) and NLRP3 (MCC950, 10 µM) (GLPBIO, USA) inhibitors were introduced to the medium 1 h in advance, respectively.
Sci Total Environ (2023): 165500. PMID: 37442457 IF: 9.8003 -
Related Biological Data
Inhibition of P2X7 alleviated LPS-induced apoptosis and pyroptosis in mRECs by attenuating the activation of P2X7 and NLRP3 inflammasome. a P2X7 and NLRP3 inflammasome protein levels.
We added A740003 (100 µmol/ml), MCC950 (100 mol/ml)(GlpBio), and 3TC (100 µmol/ml) were added to cells 6 h prior to stimulation with LPS and incubated for 72 h.
Cell Death Dis 13.4 (2022): 1-13. PMID: 35410316 IF: 8.4685 -
Related Biological Data
MCC950 attenuated NLRP3-inflammasome activation and hippocampal damage in AlCl3-treated Parkin-/- mice. (A) The expressions of NLRP3 and IL-1β proteins were detected by Western Blot (n = 3). (B) Effect of MitoQ on Al content in the hippocampus.
The second-time was treated with normal saline, MitoQ (5 mg/kg body weight, twice weekly) or MCC950 (GlpBio) (10 mg/kg body weight, twice weekly) by intraperitoneal injection in the afternoon of the same day.
Ecotox Environ Safe 264 (2023): 115459. PMID: 37703808 IF: 6.7996 -
Related Biological Data
MCC950 ameliorated IMQ-induced skin lesions in BALB/c mice by inhibiting NLRP3. (b) Mouse model of psoriasis induced in mice by topical application of IMQ. Psoriasis-like skin lesions were observed after seven days in IMQ-treated mice. However, mice treated with 20 mg/kg/d IMQ + MCC950 exhibited ameliorative symptoms.
Mice in the IMQ + MCC950 group were intra-gastrically administered with MCC950 (GLPBIO) at a dose of 20 mg/kg/d for 7 consecutive days.
Int Immunopharmacol 124 (2023): 110811. PMID: 37647679 IF: 5.5999 -
Related Biological Data
(A–B) Protein levels of NLRP3, caspase-1, and GSDMD-N (n = 3). (C) TUNEL staining of apoptosis (magnification, 400 × ). (D–E) Protein levels of Col-I and α-SMA in mice liver .
Two groups of mice (n = 6) were randomly assigned: the HFPO-TA group and the HFPO-TA + MCC950 group. Mice in the HFPO-TA + MCC950 group received intraperitoneal injections of MCC950 (10 mg/kg, twice/week for 4 weeks; GLPBIO, USA)
Food Chem Toxicol (2023): 113706. PMID: 36871880 IF: 5.5716 -
Related Biological Data
PCAinhibited the expression of pyroptosis-associated proteins.(C-I) Representative images of Western blotanalysis and the semi-quantification of NLRP3, GSDMD, Caspase-1, and IL-1β. The data are expressed as the mean ± SEM.
PCA (40 mg/kg), MCC950 (5 mg/kg)(GlpBio),and the combination treatment obviously reduced the expression of NLRP3 in the cerebral cortex under ischemic injury (P < 0.05).
Phytomedicine (2021): 153849. PMID: 34775360 IF: 5.341
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Cell experiment: |
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]. |
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Animal experiment: |
Mice[1] C57BL/6 mice are immunized subcutaneously with 150 µg of MOG peptide 35-55 emulsified in CFA containing 4 mg/mL (0.4.mg/mouse) of heat-killed MTB. Mice are injected i.p. with 500 ng pertussis toxin (PT: kaketsuken) on days 0 and 2. MCC950 is administered i.p. to mice (10 mg/kg) at induction of the disease, day 0, 1 and 2 and every 2 days thereafter. Control mice are administered vehicle (PBS) at the same time points. Mice are observed for clinical signs of disease daily (unblinded). Disease severity is scored as follows: no clinical signs, 0; limp tail, 1; ataxic gait, 2; hind limb weakness, 3; hind limb paralysis, 4; and tetra paralysis, 5. |
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References: [1]. Coll RC, et al. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat Med. 2015 Mar;21(3):248-55. |
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MCC950 is a potent and selective NLRP3 inhibitor with IC50s of 7.5 and 8.1 nM in BMDMs and HMDMs, respectively.
MCC950 blocks canonical and non-canonical NLRP3 activation at nanomolar concentrations. MCC950 specifically inhibits NLRP3 but not AIM2, NLRC4 or NLRP1 activation. The effect of MCC950 on NLRP3 inflammasome activation is tested in mouse bone marrow derived macrophages (BMDM) and human monocyte derived macrophages (HMDM). The IC50 of MCC950 in BMDM is approximately 7.5 nM, while in HMDM it has a similar inhibitory capacity (IC50=8.1 nM). MCC950 also dose dependently inhibit IL-1β but not TNF-α secretion.MCC950 specifically blocks caspase-11-directed NLRP3 activation and IL-1β secretion upon stimulation of the non-canonical pathway. NLRC4-stimulated IL-1β and TNF-α secretion (as activated by Salmonella typhimurium) are not inhibited by MCC950 even at a concentration of 10 µM. MCC950 does not inhibit caspase-1 activation or IL-1β processing in response to S. typhimurium. The expression of pro-caspase-1 and pro-IL-1β in cell lysates is not substantially affected by MCC950 treatment[1].
MCC950 reduces Interleukin-1p (IL-1β) production and attenuates the severity of experimental autoimmune encephalomyelitis (EAE), a disease model of multiple sclerosis. Pre-treatment with MCC950 reduces serum concentrations of IL-1β and IL-6 while it does not considerably decrease the amount of TNF-α. Treatment of mice with MCC950 delays the onset and reduced the severity of EAE. Intracellular cytokine staining and FACS analysis of brain mononuclear cells from mice sacrificed on day 22 shows modestly reduced frequencies of IL-17 and IFN-γ producing CD3+ T cells in MCC950 treated mice in comparison with PBS-treated mice. IFN-γ and particularly IL-17 producing cell numbers are also reduced in both the CD4+ and γδ+ sub-populations of CD3+ T cells[1].
MCC950是一种强效且选择性的NLRP3抑制剂，在BMDMs和HMDMs中的IC50分别为7.5和8.1纳摩尔。
MCC950可以在纳摩尔浓度下阻止NLRP3的典型和非典型激活。 MCC950特异性地抑制NLRP3而不是AIM2，NLRC4或NLRP1的激活。在小鼠骨髓源性巨噬细胞（BMDM）和人单核细胞衍生的巨噬细胞（HMDM）中测试了MCC950对NLRP3炎症小体激活的影响。在BMDM中，MCC950的IC50约为7.5 nM，在HMDM中具有类似的抑制能力（IC50 = 8.1 nM）。 MCC950还剂量依赖性地抑制IL-1β但不是TNF-α分泌。 MCC950特异性地阻断caspase-11定向的非经典途径刺激引起的NLRP3激活和IL-1β分泌。即使在10μm浓度下，也无法通过Salmonella typhimurium诱导NLRC4刺激引起IL-1β和TNF-α分泌来抑制MCC950。 MCC950不会通过S.typhimurium诱导抑制caspase-1活化或IL-1β加工反应。 经过McC95处理后，细胞裂解物中pro-caspase-1和pro-IL-1β的表达没有受到实质性影响[1]。
MCC950可以减少白细胞介素-1β（IL-1β）的产生，缓解实验性自身免疫脑脊髓炎（EAE）的严重程度，这是多发性硬化症的一种疾病模型。预先使用MCC950可降低血清中IL-1β和IL-6的浓度，但并不会显着降低TNF-α的数量。用MCC950治疗小鼠可以延迟EAE的发作时间，并减轻其严重程度。在第22天牺牲后对小鼠脑单个核细胞进行细胞因子染色和流式细胞分析显示，在与PBS处理组相比较下，接受MCC950治疗组CD3+ T 细胞中 IL-17 和 IFN-γ 产生频率略有降低。同时，在 CD4+ 和 γδ+ 亚群中也都减少了IFN-gamma和特别是IL-17 的产生[1]。
Reference:
[1]. Coll RC, et al. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat Med. 2015 Mar;21(3):248-55. [2]. Gan W, et al. The SGK1 inhibitor EMD638683, prevents Angiotensin II-induced cardiac inflammation and fibrosis by blocking NLRP3 inflammasome activation. Biochim Biophys Acta. 2017 Oct 3;1864(1):1-10. [3]. Zhang XY, et al. Propofol Does Not Reduce Pyroptosis of Enterocytes and Intestinal Epithelial Injury After Lipopolysaccharide Challenge. Dig Dis Sci. 2018 Jan;63(1):81-91. [4]. Qi Y, et al. NLRP3-dependent synaptic plasticity deficit in an Alzheimer’s disease amyloidosis model in vivo. Neurobiol Dis. 2018 Feb 23;114:24-30.
| Cas No. | 210826-40-7 | SDF | |
| 别名 | CP-456773; CRID3 | ||
| Canonical SMILES | O=S(C1=CC(C(C)(O)C)=CO1)(NC(NC2=C3CCCC3=CC4=C2CCC4)=O)=O | ||
| 分子式 | C20H24N2O5S | 分子量 | 404.48 |
| 溶解度 | DMSO : ≥ 28 mg/mL (69.22 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.4723 mL | 12.3616 mL | 24.7231 mL |
| 5 mM | 0.4945 mL | 2.4723 mL | 4.9446 mL |
| 10 mM | 0.2472 mL | 1.2362 mL | 2.4723 mL |
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| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases
The NOD-like receptor (NLR) family, pyrin domain-containing protein 3 (NLRP3) inflammasome is a component of the inflammatory process, and its aberrant activation is pathogenic in inherited disorders such as cryopyrin-associated periodic syndrome (CAPS) and complex diseases such as multiple sclerosis, type 2 diabetes, Alzheimer's disease and atherosclerosis. We describe the development of MCC950, a potent, selective, small-molecule inhibitor of NLRP3. MCC950 blocked canonical and noncanonical NLRP3 activation at nanomolar concentrations. MCC950 specifically inhibited activation of NLRP3 but not the AIM2, NLRC4 or NLRP1 inflammasomes. MCC950 reduced interleukin-1β (IL-1β) production in vivo and attenuated the severity of experimental autoimmune encephalomyelitis (EAE), a disease model of multiple sclerosis. Furthermore, MCC950 treatment rescued neonatal lethality in a mouse model of CAPS and was active in ex vivo samples from individuals with Muckle-Wells syndrome. MCC950 is thus a potential therapeutic for NLRP3-associated syndromes, including autoinflammatory and autoimmune diseases, and a tool for further study of the NLRP3 inflammasome in human health and disease.
NLRP3 inflammasome blockade reduces liver inflammation and fibrosis in experimental NASH in mice
Background & aims: NOD-like receptor protein 3 (NLRP3) inflammasome activation occurs in Non-alcoholic fatty liver disease (NAFLD). We used the first small molecule NLRP3 inhibitor, MCC950, to test whether inflammasome blockade alters inflammatory recruitment and liver fibrosis in two murine models of steatohepatitis.
Methods: We fed foz/foz and wild-type mice an atherogenic diet for 16weeks, gavaged MCC950 or vehicle until 24weeks, then determined NAFLD phenotype. In mice fed an methionine/choline deficient (MCD) diet, we gavaged MCC950 or vehicle for 6weeks and determined the effects on liver fibrosis.
Results: In vehicle-treated foz/foz mice, hepatic expression of NLRP3, pro-IL-1β, active caspase-1 and IL-1β increased at 24weeks, in association with cholesterol crystal formation and NASH pathology; plasma IL-1β, IL-6, MCP-1, ALT/AST all increased. MCC950 treatment normalized hepatic caspase 1 and IL-1β expression, plasma IL-1β, MCP-1 and IL-6, lowered ALT/AST, and reduced the severity of liver inflammation including designation as NASH pathology, and liver fibrosis. In vitro, cholesterol crystals activated Kupffer cells and macrophages to release IL-1β; MCC950 abolished this, and the associated neutrophil migration. MCD diet-fed mice developed fibrotic steatohepatitis; MCC950 suppressed the increase in hepatic caspase 1 and IL-1β, lowered numbers of macrophages and neutrophils in the liver, and improved liver fibrosis.
Conclusion: MCC950, an NLRP3 selective inhibitor, improved NAFLD pathology and fibrosis in obese diabetic mice. This is potentially attributable to the blockade of cholesterol crystal-mediated NLRP3 activation in myeloid cells. MCC950 reduced liver fibrosis in MCD-fed mice. Targeting NLRP3 is a logical direction in pharmacotherapy of NASH.
Lay summary: Fatty liver disease caused by being overweight with diabetes and a high risk of heart attack, termed non-alcoholic steatohepatitis (NASH), is the most common serious liver disease with no current treatment. There could be several causes of inflammation in NASH, but activation of a protein scaffold within cells termed the inflammasome (NLRP3) has been suggested to play a role. Here we show that cholesterol crystals could be one pathway to activate the inflammasome in NASH. We used a drug called MCC950, which has already been shown to block NLRP3 activation, in an attempt to reduce liver injury in NASH. This drug partly reversed liver inflammation, particularly in obese diabetic mice that most closely resembles the human context of NASH. In addition, such dampening of liver inflammation in NASH achieved with MCC950 partly reversed liver scarring, the process that links NASH to the development of cirrhosis.
Recent advances in the mechanisms of NLRP3 inflammasome activation and its inhibitors
The NLRP3 inflammasome is a multimeric protein complex that initiates an inflammatory form of cell death and triggers the release of proinflammatory cytokines IL-1β and IL-18. The NLRP3 inflammasome has been implicated in a wide range of diseases, including Alzheimer's disease, Prion diseases, type 2 diabetes, and some infectious diseases. It has been found that a variety of stimuli including danger-associated molecular patterns (DAMPs, such as silica and uric acid crystals) and pathogen-associated molecular patterns (PAMPs) can activate NLRP3 inflammasome, but the specific regulatory mechanisms of NLRP3 inflammasome activation remain unclear. Understanding the mechanisms of NLRP3 activation will enable the development of its specific inhibitors to treat NLRP3-related diseases. In this review, we summarize current understanding of the regulatory mechanisms of NLRP3 inflammasome activation as well as inhibitors that specifically and directly target NLRP3.
Structure of the NLRP3 decamer bound to the cytokine release inhibitor CRID3
NLRP3 is an intracellular sensor protein that when activated by a broad spectrum of exogenous and endogenous stimuli leads to inflammasome formation and pyroptosis1,2. The conformational states of NLRP3 and the way antagonistic small molecules act at the molecular level remain poorly understood2,3. Here we report the cryo-electron microscopy structures of full-length human NLRP3 in its native form and complexed with the inhibitor CRID3 (also named MCC950)4. Inactive, ADP-bound NLRP3 is a decamer composed of homodimers of intertwined leucine-rich repeat (LRR) domains that assemble back-to-back as pentamers. The NACHT domain is located at the apical axis of this spherical structure. One pyrin domain dimer is in addition formed inside the LRR cage. Molecular contacts between the concave sites of two opposing LRR domains are mediated by an acidic loop that extends from an LRR transition segment. Binding of CRID3 considerably stabilizes the NACHT and LRR domains relative to each other. CRID3 binds into a cleft, connecting four subdomains of the NACHT with the transition LRR. Its central sulfonylurea group interacts with the Walker A motif of the NLRP3 nucleotide-binding domain and is sandwiched between two arginine residues, which explains the specificity of NLRP3 for this chemical entity. With the determination of the binding site of this key therapeutic agent, specific targeting of NLRP3 for the treatment of autoinflammatory and autoimmune diseases and rational drug optimization is within reach.
Target of MCC950 in Inhibition of NLRP3 Inflammasome Activation: a Literature Review
MCC950 has been proposed as a specific small molecule inhibitor that can selectively block NLRP3 inflammasome activation. However, the exact mechanism of its action is still ambiguous. Accumulating investigations imply that chloride efflux-dependent ASC speck oligomerization and potassium efflux-dependent activation of caspase-1 are the two relatively independent, but indispensable events for NLRP3 inflammasome activation. Previous studies suggested that influence of MCC950 on potassium efflux and its consequent events such as interaction between NEK7 and NLRP3 are limited. However, inhibiting chloride intracellular channel-dependent chloride efflux leads to a modification of inflammatory response, which is similar to the function of MCC950. Based on these findings, we shed new insights on the understanding of MCC950 that its function might correlate with chloride efflux, chloride intracellular channels, or other targets that act upstream of chloride efflux.