Ac-YVAD-CMK
(Synonyms: Caspase-1 Inhibitor II) 目录号 : GC42721
Ac-YVAD-CMK 是一种选择性不可逆的 caspase-1 抑制剂 (Ki=0.8nM)，可阻止促炎细胞因子 IL-1β 的激活。
Cas No.:178603-78-6
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
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- Purity: >98.00%
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- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
Microglia cells |
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Preparation Method |
Microglia cells (1×105) were stimulated with the caspase-1 inhibitor Ac-YVAD-CMK (dissolved by DMSO) for 1h. Thrombin was added at a final concentration of 10U/mL to activate the microglia cells for 24h. |
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Reaction Conditions |
40μmol/L or 80μmol/L |
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Applications |
High level Ac-YVAD-CMK treatment (40μmol/L or 80μmol/L) significantly decreased the mRNA levels of IL-1β/IL-18. The 40μmol/L dose of Ac-YVAD-CMK significantly reduced the protein levels of caspase-1 (p20) and mature IL-1β/IL-18 compared with the thrombin-activated microglia group. |
| Animal experiment [2]: | |
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Animal models |
Male Sprague Dawley rats，weighing 250-275g |
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Preparation Method |
Ac-YVAD-CMK (300ng/rat in 3mL) was injected intracerebroventricularly 10 min after permanent middle cerebral artery occlusion in the rat. |
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Dosage form |
300ng/rat in 3mL, intracerebroventricular injection |
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Applications |
Ac-YVAD-CMK treatment induced a significant reduction of infarct volume not only 24 h after ischemia but also 6d later. Ac-YVAD-CMK treatment resulted in a reduction not only of caspase-1 but also of caspase-3 activity at 24h and led to a parallel decrease of apoptosis as measured by nucleosome quantitation. Likewise, brain levels of the proinflammatory cytokines IL-1b and TNF-a were reduced at 24h. |
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References: [1]. Liang H, Sun Y, et al. Ac-YVAD-cmk improves neurological function by inhibiting caspase-1-mediated inflammatory response in the intracerebral hemorrhage of rats. Int Immunopharmacol. 2019;75:105771. [2]. Rabuffetti M, Sciorati C, et al. Inhibition of caspase-1-like activity by Ac-Tyr-Val-Ala-Asp-chloromethyl ketone induces long-lasting neuroprotection in cerebral ischemia through apoptosis reduction and decrease of proinflammatory cytokines. J Neurosci. 2000;20(12):4398-4404. |
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Ac-YVAD-CMK is a selective irreversible inhibitor of caspase-1 (Ki=0.8nM), which can prevent the proinflammatory cytokine IL-1β activation. Ac-YVAD-CMK can reduce the inflammatory response and induce a long-lasting neuroprotective effect[1,2,3].
Ac-YVAD-CMK reduced the expression of IL-1β and IL-18 in activated microglia in vitro[3]. Ac-YVAD-CMK(40μM) decreased the rapid cell (PAMs, 3D4/21 cells and the endothelial cell line) death induced by ApxⅠ[4]. Ac-YVAD-CMK attenuated the inhibitory effects of berberine on the viability, migration and invasion of HepG2 cells[5].
AC-YVAD-CMK treatment significantly alleviated sepsis-induced acute kidney injury, with decreased histological injury in renal tissues, suppressed the accumulation of neutrophils and macrophages in renal tissues, and decreased sCR and BUN level[6]. Ac-YVAD-CMK protected the brain against ICH-induced injury, and the neuroprotective effect may result from anti-inflammation-induced blood–brain barrier protection[7]. Pretreatment of rats with Ac-YVAD-CMK (12.5mM/kg) significantly reduced endotoxin-induced mortality from 83% to 33% using Log Rank analysis[8].
References:
[1] Wang F, Li G, et al. Alcohol accumulation promotes esophagitis via pyroptosis activation. Int J Biol Sci. 2018;14(10):1245-1255. Published 2018 Jul 13.
[2] Rabuffetti M, Sciorati C, et al. Inhibition of caspase-1-like activity by Ac-Tyr-Val-Ala-Asp-chloromethyl ketone induces long-lasting neuroprotection in cerebral ischemia through apoptosis reduction and decrease of proinflammatory cytokines. J Neurosci. 2000;20(12):4398-4404.
[3] Liang H, Sun Y, et al. Ac-YVAD-cmk improves neurological function by inhibiting caspase-1-mediated inflammatory response in the intracerebral hemorrhage of rats. Int Immunopharmacol. 2019;75:105771.
[4] Hernandez-Cuellar E, Guerrero-Barrera AL, et al. An in vitro study of ApxI from Actinobacillus pleuropneumoniae serotype 10 and induction of NLRP3 inflammasome-dependent cell death. Vet Rec Open. 2021;8(1):e20. Published 2021 Oct 4.
[5] Chu Q, Jiang Y, et al. Pyroptosis is involved in the pathogenesis of human hepatocellular carcinoma. Oncotarget. 2016;7(51):84658-84665.
[6] Yang M, Fang JT, et al. Caspase-1-Inhibitor AC-YVAD-CMK Inhibits Pyroptosis and Ameliorates Acute Kidney Injury in a Model of Sepsis. Biomed Res Int. 2021;2021:6636621.
[7] Wu B, Ma Q, et al. Ac-YVAD-CMK Decreases Blood-Brain Barrier Degradation by Inhibiting Caspase-1 Activation of Interleukin-1β in Intracerebral Hemorrhage Mouse Model. Transl Stroke Res. 2010;1(1):57-64.
[8] Mathiak G, Grass G, et al. Caspase-1-inhibitor ac-YVAD-cmk reduces LPS-lethality in rats without affecting haematology or cytokine responses. Br J Pharmacol. 2000;131(3):383-386.
Ac-YVAD-CMK 是一种选择性不可逆的 caspase-1 抑制剂 (Ki=0.8nM)，可阻止促炎细胞因子 IL-1β 的激活。 Ac-YVAD-CMK 可降低炎症反应并诱导持久的神经保护作用[1,2,3]。
Ac-YVAD-CMK 在体外降低活化小胶质细胞中 IL-1β 和 IL-18 的表达[3]。 Ac-YVAD-CMK(40μM)降低ApxⅠ诱导的快速细胞(PAMs、3D4/21细胞和内皮细胞系)死亡[4]。 Ac-YVAD-CMK减弱小檗碱对HepG2细胞活力、迁移和侵袭的抑制作用[5]。
AC-YVAD-CMK 治疗显着减轻脓毒症引起的急性肾损伤，减少肾组织的组织学损伤，抑制肾组织中中性粒细胞和巨噬细胞的积累，降低 sCR 和 BUN 水平[6]< /sup>。 Ac-YVAD-CMK 保护大脑免受 ICH 诱导的损伤，神经保护作用可能来自抗炎诱导的血脑屏障保护[7]。使用对数秩分析[8]，用 Ac-YVAD-CMK (12.5mM/kg) 预处理大鼠可将内毒素诱导的死亡率从 83% 显着降低至 33%。
| Cas No. | 178603-78-6 | SDF | |
| 别名 | Caspase-1 Inhibitor II | ||
| 化学名 | N-acetyl-L-tyrosyl-L-valyl-N-[(1S)-1-(carboxymethyl)-3-chloro-2-oxo-propyl]-L-alaninamide | ||
| Canonical SMILES | ClCC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)C)Cc1ccc(O)cc1)C(C)C | ||
| 分子式 | C24H33ClN4O8 | 分子量 | 541 |
| 溶解度 | 20mg/mL in DMSO, 10mg/mL in DMF | 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 | 1.8484 mL | 9.2421 mL | 18.4843 mL |
| 5 mM | 0.3697 mL | 1.8484 mL | 3.6969 mL |
| 10 mM | 0.1848 mL | 0.9242 mL | 1.8484 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 | ||||||||||
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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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Caspase-1-Inhibitor Ac-YVAD-CMK Inhibits Pyroptosis and Ameliorates Acute Kidney Injury in a Model of Sepsis
Biomed Res Int 2021 Jun 10;2021:6636621.PMID:34222479DOI:10.1155/2021/6636621.
Objective: To observe the protective effect of Ac-YVAD-CMK on sepsis-induced acute kidney injury in mice and to explore its possible mechanisms primarily. Methods: Eighteen male C57BL/6 mice were randomly divided into sham-operated group (Control), cecal ligation and puncture group (CLP), and CLP model treated with Ac-YVAD-CMK group (Ac-YVAD-CMK) (n = 6 in each group). Mice were sacrificed at 24 h after operation, and blood and kidney tissue samples were collected for analyses. Histologic changes were determined microscopically following HE staining. The expression of Ly-6B and CD68 was investigated using immunohistochemistry. Serum concentrations of creatinine (sCR) and blood urea nitrogen (BUN) were measured. Serum levels of interleukin-1β (IL-1β), interleukin-18 (IL-18), TNF-α, and interleukin-6 (IL-6) were determined by ELISA. The expressions of Caspas-1, NLRP-1, IL-1β, and IL-18 in renal tissues were investigated using Western blot. Immunofluorescence staining was used to detect the expression of GSDMD protein in renal tissues. Results: Ac-YVAD-CMK treatment significantly alleviates sepsis-induced acute kidney injury, with decreased histological injury in renal tissues, suppresses the accumulation of neutrophils and macrophages in renal tissues, and decreased sCR and BUN level (P < 0.05). Attenuation of sepsis-induced acute kidney injury was due to the prohibited production of inflammatory cytokines and decrease expression of Caspas-1, NLRP-1, IL-1β, and IL-18 in renal tissues. In addition, Ac-YVAD-CMK treatment significantly reduced the expression of GSDMD in renal tissues compared to those observed in controls (P < 0.05). Conclusions: We demonstrated a marked renoprotective effect of caspase-1-inhibitor Ac-YVAD-CMK in a rat model of sepsis by inhibition of pyroptosis.
Lipopolysaccharide (LPS) Aggravates High Glucose- and Hypoxia/Reoxygenation-Induced Injury through Activating ROS-Dependent NLRP3 Inflammasome-Mediated Pyroptosis in H9C2 Cardiomyocytes
J Diabetes Res 2019 Feb 17;2019:8151836.PMID:30911553DOI:10.1155/2019/8151836.
Diabetes aggravates myocardial ischemia-reperfusion (I/R) injury because of the combination effects of changes in glucose and lipid energy metabolism, oxidative stress, and systemic inflammatory response. Studies have indicated that myocardial I/R may coincide and interact with sepsis and inflammation. However, the role of LPS in hypoxia/reoxygenation (H/R) injury in cardiomyocytes under high glucose conditions is still unclear. Our objective was to examine whether lipopolysaccharide (LPS) could aggravate high glucose- (HG-) and hypoxia/reoxygenation- (H/R-) induced injury by upregulating ROS production to activate NLRP3 inflammasome-mediated pyroptosis in H9C2 cardiomyocytes. H9C2 cardiomyocytes were exposed to HG (30 mM) condition with or without LPS, along with caspase-1 inhibitor (Ac-YVAD-CMK), inflammasome inhibitor (BAY11-7082), ROS scavenger N-acetylcysteine (NAC), or not for 24 h, then subjected to 4 h of hypoxia followed by 2 h of reoxygenation (H/R). The cell viability, lactate dehydrogenase (LDH) release, caspase-1 activity, and intracellular ROS production were detected by using assay kits. The incidence of pyroptosis was detected by calcein-AM/propidium iodide (PI) double staining kit. The concentrations of IL-1β and IL-18 in the supernatants were assessed by ELISA. The mRNA levels of NLRP3, ASC, and caspase-1 were detected by qRT-PCR. The protein levels of NF-κB p65, NLRP3, ASC, cleaved caspase-1 (p10), IL-1β, and IL-18 were detected by western blot. The results indicated that pretreatment LPS with 1 μg/ml not 0.1 μg/ml could efficiently aggravate HG and H/R injury by activating NLRP3 inflammasome to mediate pyroptosis in H9C2 cells, as evidenced by increased LDH release and decreased cell viability in the cells, and increased expression of NLRP3, ASC, cleaved caspase-1 (p10), IL-1β, and IL-18. Meanwhile, Ac-YVAD-CMK, BAY11-7082, or NAC attenuated HG- and H/R-induced H9C2 cell injury with LPS stimulated by reversing the activation of NLRP3 inflammasome-mediated pyroptosis. In conclusion, LPS could increase the sensitivity of H9C2 cells to HG and H/R and aggravated HG- and H/R-induced H9C2 cell injury by promoting ROS production to induce NLRP3 inflammasome-mediated pyroptosis.
Ac-YVAD-CMK Inhibits Pyroptosis and Improves Functional Outcome after Intracerebral Hemorrhage
Biomed Res Int 2018 Oct 16;2018:3706047.PMID:30410928DOI:10.1155/2018/3706047.
Intracerebral hemorrhage (ICH) refers to bleeding in the brain and is associated with the release of large amount of inflammasomes, and the activation of different cell death pathways. These cell death pathways lead to removal of inactivated and damaged cells and also result in neuronal cell damage. Pyroptosis is a newly discovered cell death pathway that has gained attention in recent years. This pathway mainly depends on activation of caspase-1-mediated cascades to cause cell death. We tested a well-known selective inhibitor of caspase-1, Ac-YVAD-CMK, which has previously been found to have neuroprotective effects in ICH mice model, to ascertain its effects on the activation of inflammasomes mediated pyroptosis. Our results showed that Ac-YVAD-CMK could reduce caspase-1 activation and inhibit IL-1β production and maturation, but has no effect on NLRP3 expression, an upstream inflammatory complex. Ac-YVAD-CMK administration also resulted in reduction in M1-type microglia polarization around the hematoma, while increasing the number of M2-type cells. Furthermore, Ac-YVAD-CMK treated mice showed some recovery of neurological function after hemorrhage especially at the hyperacute and subacute stage resulting in some degree of limb movement. In conclusion, we are of the view that Ac-YVAD-CMK could inhibit pyroptosis, decrease the secretion or activation of inflammatory factors, and affect the polarization of microglia resulting in improvement of neurological function after ICH.
SARS-CoV-2 N protein promotes NLRP3 inflammasome activation to induce hyperinflammation
Nat Commun 2021 Aug 2;12(1):4664.PMID:34341353DOI:10.1038/s41467-021-25015-6.
Excessive inflammatory responses induced upon SARS-CoV-2 infection are associated with severe symptoms of COVID-19. Inflammasomes activated in response to SARS-CoV-2 infection are also associated with COVID-19 severity. Here, we show a distinct mechanism by which SARS-CoV-2 N protein promotes NLRP3 inflammasome activation to induce hyperinflammation. N protein facilitates maturation of proinflammatory cytokines and induces proinflammatory responses in cultured cells and mice. Mechanistically, N protein interacts directly with NLRP3 protein, promotes the binding of NLRP3 with ASC, and facilitates NLRP3 inflammasome assembly. More importantly, N protein aggravates lung injury, accelerates death in sepsis and acute inflammation mouse models, and promotes IL-1β and IL-6 activation in mice. Notably, N-induced lung injury and cytokine production are blocked by MCC950 (a specific inhibitor of NLRP3) and Ac-YVAD-CMK (an inhibitor of caspase-1). Therefore, this study reveals a distinct mechanism by which SARS-CoV-2 N protein promotes NLRP3 inflammasome activation and induces excessive inflammatory responses.
Ac-YVAD-CMK improves neurological function by inhibiting caspase-1-mediated inflammatory response in the intracerebral hemorrhage of rats
Int Immunopharmacol 2019 Oct;75:105771.PMID:31352322DOI:10.1016/j.intimp.2019.105771.
Objective: Intracerebral hemorrhage (ICH) is acknowledged as a serious clinical problem lacking effective treatments. And caspase-1-mediated inflammatory response happened during the progression of ICH. Therefore, we aimed to investigate the effects of caspase-1 inhibitor Ac-YVAD-CMK on ICH. Materials and methods: Microglia cells were isolated and activated by thrombin for 24 h. Then the transcript and protein expressions of NLRP3 and inflammatory factors were assessed by RT-PCR and western blotting. Moreover, Ac-YVAD-CMK was injected into the ICH model. The mNSS and brain water content were tested at 24 h post-ICH. Finally, the pathological changes of microglia activation following ICH were discovered by the immunohistochemical and HE staining ways. Results: Ac-YVAD-CMK inhibited the activation of pro-caspase-1 and decreased brain edema, in association with decreasing activated microglia and the expression of inflammation-related factors at 24 h post-ICH. Consequently, Ac-YVAD-CMK reduced the release of mature IL-1β/IL-18 in perihematoma, improved the behavioral performance, and alleviated microglia in perihematoma region in ICH rats. Conclusions: These results indicate that caspase-1 could amplify the plural inflammatory responses in the ICH. Administration of Ac-YVAD-CMK has the potential to be a novel therapeutic strategy for ICH.