CY-09
(Synonyms: 4-[[2-硫代-3-(3-三氟甲基苄基)-4-氧代噻唑烷-5-亚基]甲基]苯甲酸) 目录号 : GC19113
An NLRP3 inhibitor
Cas No.:1073612-91-5
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Kinase experiment [1]: | |
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Preparation Method |
For ATPase activity assay, purified recombinant human proteins (1.4 ng/uL) were incubated at 37°C with indicated concentrations of CY-09 for 15 min in the reaction buffer. ATP (25 um, Ultra-Pure ATP) was then added, and the mixture was further incubated at 37 °C for another 40 min. |
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Reaction Conditions |
0.1-1uM CY-09 with protein incubate for 15 minutes |
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Applications |
Cy-09 inhibited the ATPase activity of purified NLRP3 at a dose of 0.1 - 1 uM. The inhibitory effect of CY-09 on NLRP3 ATPase activity was specific because it had no effect on the ATPase activity of purified NLRC4 NLRP1 NOD2 or RIG-I. |
| Cell experiment [1]: | |
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Cell lines |
MDM cells(BMDM and PMDM) |
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Preparation Method |
To induce NLRP3 inflammasome activation, MDM cells were stimulated with LPS for 3 h. CY-09 or other inhibitors were added into the culture for 30 min, and then the cells were stimulated for 4 h with MSU, Salmonella typhimurium or for 30 min with ATP or nigericin. |
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Reaction Conditions |
1-10 µM Cy-09 for 30 min |
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Applications |
Cy-09 specifically blocks NLRP3 activation in macrophages. |
| Animal experiment [1]: | |
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Animal models |
C57BL/6J mice |
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Preparation Method |
C57BL/6J mice were injected with 40 mg/kg CY-09 or vehicle 30 min before injection of MSU. After 6 h, mice were killed, and peritoneal cavities underwent lavage with 10 ml ice-cold PBS. |
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Dosage form |
40 mg/kg CY-09 (intraperitoneal injection) |
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Applications |
Cy-09 inhibited NLRP3 activation in vivo and prevented neonatal mortality in CAPS mouse model. |
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References: [1]. Jiang H, He H, et,al.Identification of a selective and direct NLRP3 inhibitor to treat inflammatory disorders. J Exp Med. 2017 Nov 6;214(11):3219-3238. doi: 10.1084/jem.20171419. Epub 2017 Oct 11. PMID: 29021150; PMCID: PMC5679172. |
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CY-09, a specific inhibitor of the NLRP3 inflammasome, directly targets NLRP3 and has an IC50 value of 18.9, 8.18, >50, >50 and 26.0 uM for each of the five major cytochrome P450 enzymes. CY-09 directly binds to the ATP-binding motif of NLRP3 NACHT domain and inhibits NLRP3 ATPase activity, resulting in the suppression of NLRP3 inflammasome assembly and activation[1].
CY-09 exhibited a dose-dependent inhibitory effect on monosodium urate (MSU), nigericin, and ATP-induced caspase-1 activation and IL-1β secretion at the doses of 1-10 uM in LPS-primed BMDMs. Cytosolic LPS induced noncanonical NLRP3 activation in BMDMs could also be blocked by CY-09 treatment[1]. CY-09 can maintain extracellular matrix (ECM) homeostasis and regulate inflammation in TNF-α treated chondrocytes via inhibition of NLRP3 inflammasome-mediated pyroptosis[2].CY-09 reduced the production of inflammatory cytokines, intracellular Ca2+ levels, and the activation of TRPA1 by inhibiting the activation of inflammasomes, thereby reducing the proinflammatory polarization of macrophages and alleviating animal pain and injury[3]. M1-polarized macrophage infiltration and NLRP3 inflammasome activation increased in the cardiac ventricle after diabetic stroke. CY-09 restored cardiac function, The M1-polarized macrophage-NLRP3 inflammasome activation is a pathway underlying the brain-heart interaction after diabetic stroke[7].
CY-09 was tested against the five major cytochrome P450 enzymes 1A2, 2C9, 2C19, 2D6, and 3A4 with half maximal inhibitory concentration (IC50) values of 18.9, 8.18, >50, >50, and 26.0 uM, respectively, which exhibited low risk of drug drug interactions. CY-09 treatment in vivo efficiently suppressed MSU injection induced IL-1β production and neutrophil influx, suggesting that CY-09 can block MSU-induced NLRP3 inflammasome activation in vivo[1]. When investigate whether CY-09 is effective for the treatment of NAFLD in a high-fat diet (HFD)-induced mouse model.CY-09 reduces hepatic steatosis in experimental NAFLD mice and CY-09 may be a potential therapeutic drug of NAFLD in clinical practice[4]. Compared with surgery alone, sleeve gastroplasty mimicking ESG combined with CY-09 resulted in weight loss, significantly improved insulin resistance, and better remission of NAS[5]. In db/db mice, inflammation, oxidative stress, apoptosis and fibrosis increased, while CY-09 exerted renoprotection by inhibiting NLRP3 inflammasome activation[6].
References:
[1]: Jiang H, He H, et,al. Identification of a selective and direct NLRP3 inhibitor to treat inflammatory disorders. J Exp Med. 2017 Nov 6;214(11):3219-3238. doi: 10.1084/jem.20171419. Epub 2017 Oct 11. PMID: 29021150; PMCID: PMC5679172.
[2]: Zhang Y, Lin Z, et,al. CY-09 attenuates the progression of osteoarthritis via inhibiting NLRP3 inflammasome-mediated pyroptosis. Biochem Biophys Res Commun. 2021 May 14;553:119-125. doi: 10.1016/j.bbrc.2021.03.055. Epub 2021 Mar 22. PMID: 33765556.
[3]: Fan Y, Xue G, et,al. CY-09 Inhibits NLRP3 Inflammasome Activation to Relieve Pain via TRPA1. Comput Math Methods Med. 2021 Aug 14;2021:9806690. doi: 10.1155/2021/9806690. PMID: 34426748; PMCID: PMC8380162.
[4]: Wang X, Sun K, et,al. NLRP3 inflammasome inhibitor CY-09 reduces hepatic steatosis in experimental NAFLD mice. Biochem Biophys Res Commun. 2021 Jan 1;534:734-739. doi: 10.1016/j.bbrc.2020.11.009. Epub 2020 Nov 16. PMID: 33213837.
[5]: Sun K, Wang J, et,al. Sleeve Gastroplasty Combined with the NLRP3 Inflammasome Inhibitor CY-09 Reduces Body Weight, Improves Insulin Resistance and Alleviates Hepatic Steatosis in Mouse Model. Obes Surg. 2020 Sep;30(9):3435-3443. doi: 10.1007/s11695-020-04571-8. PMID: 32266697.
[6]: Yang M, Zhao L. The selective NLRP3-inflammasome inhibitor CY-09 ameliorates kidney injury in diabetic nephropathy by inhibiting NLRP3 inflammasome activation. Curr Med Chem. 2022 Sep 22. doi: 10.2174/0929867329666220922104654. Epub ahead of print. PMID: 36154582.
[7]: Lin HB, Wei GS, et,al. Macrophage-NLRP3 Inflammasome Activation Exacerbates Cardiac Dysfunction after Ischemic Stroke in a Mouse Model of Diabetes. Neurosci Bull. 2020 Sep;36(9):1035-1045. doi: 10.1007/s12264-020-00544-0. Epub 2020 Jul 18. PMID: 32683554; PMCID: PMC7475163.
CY-09 是 NLRP3 炎性体的特异性抑制剂，直接靶向 NLRP3，对五种主要细胞色素 P450 酶中的每一种酶的 IC50 值为 18.9、8.18、>50、>50 和 26.0 uM。 CY-09 直接结合 NLRP3 NACHT 结构域的 ATP 结合基序并抑制 NLRP3 ATP 酶活性，从而抑制 NLRP3 炎性体组装和激活[1]。
CY-09 在 LPS 引发的 BMDM 中以 1-10 uM 的剂量对尿酸单钠 (MSU)、尼日利亚菌素和 ATP 诱导的 caspase-1 激活和 IL-1β 分泌表现出剂量依赖性抑制作用。 CY-09 处理也可阻断 BMDM 中胞质 LPS 诱导的非经典 NLRP3 激活[1]。 CY-09 可通过抑制 NLRP3 炎性体介导的细胞焦亡来维持细胞外基质 (ECM) 稳态并调节 TNF-α 处理的软骨细胞的炎症[2]。CY-09 减少炎症细胞因子的产生，细胞内Ca2+ 水平，以及 TRPA1 的激活，通过抑制炎性体的激活，从而减少巨噬细胞的促炎性极化，减轻动物的疼痛和损伤[3]。糖尿病卒中后心室中 M1 极化巨噬细胞浸润和 NLRP3 炎性体激活增加。 CY-09恢复心脏功能，M1极化巨噬细胞-NLRP3炎性体激活是糖尿病脑卒中后脑-心相互作用的潜在通路[7]。
CY-09 针对五种主要细胞色素 P450 酶 1A2、2C9、2C19、2D6 和 3A4 进行了测试，半数最大抑制浓度 (IC50) 值为 18.9、8.18、>50 , >50 和 26.0 uM，分别表现出药物相互作用的低风险。 CY-09 体内治疗有效抑制 MSU 注射诱导的 IL-1β 产生和中性粒细胞流入，表明 CY-09 可以阻断 MSU 诱导的体内 NLRP3 炎性体激活[1]。当研究 CY-09 在高脂饮食 (HFD) 诱导的小鼠模型中是否对治疗 NAFLD 有效时。CY-09 减少实验性 NAFLD 小鼠的肝脂肪变性，CY-09 可能是 NAFLD 的潜在治疗药物。临床实践[4].与单纯手术相比，模拟ESG的袖状胃成形术联合CY-09可减轻体重，显着改善胰岛素抵抗，更好地缓解NAS[5]。在db/db小鼠中，炎症、氧化应激、细胞凋亡和纤维化增加，而CY-09通过抑制NLRP3炎性体激活发挥肾脏保护作用[6]。
| Cas No. | 1073612-91-5 | SDF | |
| 别名 | 4-[[2-硫代-3-(3-三氟甲基苄基)-4-氧代噻唑烷-5-亚基]甲基]苯甲酸 | ||
| Canonical SMILES | OC(C1=CC=C(/C=C2SC(N(CC3=CC=CC(C(F)(F)F)=C3)C\2=O)=S)C=C1)=O | ||
| 分子式 | C19H12F3NO3S2 | 分子量 | 423.43 |
| 溶解度 | DMSO : ≥ 150 mg/mL (354.25 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 | 2.3617 mL | 11.8083 mL | 23.6167 mL |
| 5 mM | 0.4723 mL | 2.3617 mL | 4.7233 mL |
| 10 mM | 0.2362 mL | 1.1808 mL | 2.3617 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 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 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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CY-09 Alleviates the Depression-like Behaviors via Inhibiting NLRP3 Inflammasome-Mediated Neuroinflammation in Lipopolysaccharide-Induced Mice
Depression is a serious mental illness, mainly characterized as large mood swings and sleep, diet, and cognitive function disorders. NLPR3, one of the inflammasomes that can be activated by a variety of stimuli to promote the maturation and secretion of pro-inflammatory cytokines, has been considered to be involved in the pathophysiology of depression. In this study, the putative role of CY-09, a selective and direct inhibitor of NLRP3, was evaluated in the lipopolysaccharide (LPS)-induced mice. The results of the study indicated that CY-09 significantly decreased the levels of NLRP3 in the hippocampus of LPS-induced mice. In addition, CY-09 increased the sucrose preference and shortened the immobility time in LPS-induced mice, suggesting the antidepressant-like effects of inhibiting NLRP3 inflammasome. Biochemical analysis showed that LPS significantly activated the NLRP3/ASC/cytokine signaling pathway and caused microglial activation, while CY-09 prevented the changes. Moreover, CY-09 increased the brain-derived neurotrophic factor (BDNF) only in microglia but not in the whole hippocampus. Meanwhile, CY-09 did not promote neurogenesis in the hippocampus of LPS mice. In conclusion, the results of the study showed that the antidepressant-like effects of NLRP3 inhibitor CY-09 were mediated by alleviating neuroinflammation in microglia and independent of the neurotrophic function in the hippocampus.
CY-09 Inhibits NLRP3 Inflammasome Activation to Relieve Pain via TRPA1
Peripheral tissue damage leads to inflammatory pain, and inflammatory cytokine releasing is the key factor for inducing the sensitization of nociceptors. As a calcium ion channel, TRPA1 plays an important role in pain and inflammation, thus becoming a new type of anti-inflammatory and analgesic target. However, there is no consensus on the role of this channel in mechanical hyperalgesia caused by inflammation. Here, we aim to explore the role and underlying mechanism of the inflammasome inhibitor CY-09 in two classic inflammatory pain models. We evaluated pain behavior on animal models, cytokine levels, intracellular Ca2+ levels, transient TRPA1 expression, NF-百B transcription, and NLPR3 inflammasome activation. Consistently, CY-09 reduced the production of inflammatory cytokines, intracellular Ca2+ levels, and the activation of TRPA1 by inhibiting the activation of inflammasomes, thereby reducing the proinflammatory polarization of macrophages and alleviating animal pain and injury. Importantly, AITC (TRPA1 agonist) significantly reversed the analgesic effect of CY-09, indicating that TRPA1 was involved in the analgesic effect of CY-09. Our findings indicate that CY-09 relieves inflammation and pain via inhibiting TRPA1-mediated activation of NLRP3 inflammasomes. Thus, NLRP3 inflammasome may be a potential therapeutic target for pain treatment and CY-09 may be a pharmacological agent to relieve inflammatory pain, which needs further research.
CY-09 attenuates the progression of osteoarthritis via inhibiting NLRP3 inflammasome-mediated pyroptosis
Excessive activation of inflammation in chondrocyte has been considered to be a major reason cause of cellular death and degeneration in osteoarthritis (OA) development. The NLRP3 inflammasome-mediated pyroptosis pathway is closely related to inflammation regulation. This research was conducted to confirm whether NLRP3 expression and activity are impacted in the development of OA and to detect the role of CY-09, a selective and direct inhibitor of NLRP3 in the in vitro and in vivo models of OA. Our findings corroborated that the expression of NLRP3 is stimulated in OA cartilage. CY-09 can maintain extracellular matrix (ECM) homeostasis and regulate inflammation in TNF-汐 treated chondrocytes via inhibition of NLRP3 inflammasome-mediated pyroptosis. Moreover, the chondrocyte protective effects of CY-09 were further confirmed in vivo in a DMM-induced OA model. In conclusion, our research indicates that experimental OA activated the NLRP3 activity, and pharmacological inhibition of NLRP3 inflammasome activation by CY-09 protects chondrocytes against inflammation and attenuates OA development.
NLRP3 inflammasome inhibitor CY-09 reduces hepatic steatosis in experimental NAFLD mice
Aims: Non-alcoholic fatty liver disease (NAFLD) has become one of the most common chronic liver diseases. The NOD-like receptor protein 3 (NLRP3) inflammasome was suggested to be involved in the pathogenesis of NAFLD. A small-molecule named CY-09 is a new selective and direct inhibitor of the NLRP3 inflammasome. We aimed to investigate whether CY-09 is effective for the treatment of NAFLD in a high-fat diet (HFD)-induced mouse model.
Methods: Twenty mice were fed by HFD for 14 weeks, and then were randomly assigned into two groups: (1) control group receiving dimethylsulfoxide (DMSO) solution; (2) CY-09 group receiving CY-09 injection. In an 8-week follow-up, oral glucose tolerance test (OGTT) and homeostasis model assessment of insulin resistance (HOMA-IR) were used to measure glucose metabolism. Liver steatosis was evaluated by the NAFLD activity score (NAS) and deemed as the primary outcome.
Results: The body weight in CY-09 group was significantly lower than the DMSO control group on 27 weeks (41.0 ㊣ 3.5 g vs. 49.7 ㊣ 5.2 g, P = 0.014). The area under the curve (AUC) of OGTT was less in CY-09 group than that in DMSO group (35.81 ㊣ 6.79 vs. 22.91 ㊣ 2.58 mmol/L﹞hr, P = 0.004), as well as HOMA-IR (14.36 ㊣ 3.89 vs. 8.82 ㊣ 2.04 mmol.mIU.L-2, P = 0.023). Microscopically, liver lipid droplets dramatically improved and significantly lower NAS was observed in CY-09 group (8.25 ㊣ 1.26 vs. 3.20 ㊣ 0.45, P < 0.001).
Conclusion: CY-09 reduces hepatic steatosis in experimental NAFLD mice and CY-09 may be a potential therapeutic drug of NAFLD in clinical practice.
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-1汕 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.