R-7050
(Synonyms: TNF-α Antagonist III) 目录号 : GC11659
R-7050 是一种新型细胞渗透性三唑并喹喔啉化合物，可选择性抑制 TNF-α；诱导细胞信号传导。
Cas No.:303997-35-5
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
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- Purity: >99.00%
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- SDS (Safety Data Sheet)
- Datasheet
| Kinase experiment [1]: | |
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Preparation Method |
A549 cells plated. The next day, compound library(including R-7050) prepared were added at a final concentration of 10 μM in 0.2% (v/v) DMSO. Cells were preincubated with compounds for 1 hr prior to a 4 hr stimulation with either TNFα or IL-1β. |
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Reaction Conditions |
10 μM R-7050 and cell for 1h |
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Applications |
In TNFα-induced ICAM-1 expression, R-7050 inhibition potency (EC50 = 0.63 μM) was 2- to 3-fold greater than EC50 for IL-1β-induced ICAM-1 expression (1.45 μM). |
| Cell experiment [2]: | |
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Cell lines |
Wild-type RBL-2H3 cells |
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Preparation Method |
Wherever indicated, 6.7 μM of R-7050 (dissolved in DMSO) or 0.1 μM of bafilomycin A1 was added along with anti-TNP IgE during sensitization. |
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Reaction Conditions |
6.7 μM R-7050 |
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Applications |
R-7050 significantly reduced TNF proliferation when autophagy was also inhibited by Baf-A1. |
| Animal experiment [3]: | |
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Animal models |
Male CD-1 mice (8–10 weeks old) |
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Preparation Method |
R-7050 (6–18 mg/kg) was administered via intraperitoneal route at the time of injury or up to 2 h post-ICH. |
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Dosage form |
6–18 mg/kg R-7050 for 0.5-2h |
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Applications |
R-7050 attenuates neurovascular injury after ICH. |
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References: [1]. Gururaja TL, Yung S, Ding R, Huang J, Zhou X, McLaughlin J, Daniel-Issakani S, Singh R, Cooper RD, Payan DG, Masuda ES, Kinoshita T. A class of small molecules that inhibit TNFalpha-induced survival and death pathways via prevention of interactions between TNFalphaRI, TRADD, and RIP1. Chem Biol. 2007 Oct;14(10):1105-18. doi: 10.1016/j.chembiol.2007.08.012. PMID: 17961823. [2]. Ayo TE, Adhikari P, et,al. TNF Production in Activated RBL-2H3 Cells Requires Munc13-4. Inflammation. 2020 Apr;43(2):744-751. doi: 10.1007/s10753-019-01161-4. PMID: 31897916; PMCID: PMC7176528. [3]. King MD, Alleyne CH Jr, Dhandapani KM. TNF-alpha receptor antagonist, R-7050, improves neurological outcomes following intracerebral hemorrhage in mice. Neurosci Lett. 2013 May 10;542:92-6. doi: 10.1016/j.neulet.2013.02.051. Epub 2013 Mar 7. PMID: 23499961; PMCID: PMC3744337. |
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R-7050 is a novel cell-permeable triazoloquinoxaline compound that selectively inhibited TNF-α induced cellular signaling[2].
In RBL-2H3 cells R-7050 significantly reduced TNF proliferation when autophagy was also inhibited by Baf-A1[7].CA to reduce cell viability in MDA-MB-231 breast cancer cells and tumorigenic HEK 293 cells but not in normal NIH3T3 fibroblast cells. Abundance of TNFA, TNF Receptor 1 (TNFR1) and cleaved caspase-8/-3 proapoptotic proteins to increase with CA treatment. Blocking of TNFA-TNFR1 signalling by small molecule inhibitor, R-7050, reduced the expression of cleaved caspase-8 and caspase-3 at the protein level[3]. TRPA1 channel was positively expressed in the cell bodies and processes of HODs. The expression TRPA1 channel was significantly up-regulated by high concentration of TNF-α, which could be suppressed by R-7050[5].
A single administration of R-7050 reduced neurovascular injury after ICH. In contrast to biologic approaches that directly bind and neutralize TNF-α activity, R-7050 selectively inhibits the association of TNFR with intracellular adaptor molecules, such as TRADD and RIP1, limiting receptor internalization and preventing subsequent cellular responses after TNF-α binding[1]. Capsaicin, a selective TRPV1 agonist, increased ipsilateral afferent renal nerve activity in WT but not TRPV1-/- mice. WD intake increased leptin, IL-6, and TNF-α in adipose tissue, and TNF-α antagonist III, R-7050, decreased leptin in TRPV1-/--WD. The urinary albumin level was higher in TRPV1-/--WD than WT-WD[4]. Treatment with the TNF receptor-1 inhibitor R-7050 during nephrotoxic serum nephritis reduced damage, fibrosis, and necroptosis in wild-type mice and mice with KLF4-deficient macrophages, and abrogated the differences between the two groups in these parameters[6].
References:
[1]. King MD, Alleyne CH Jr,et,al. TNF-alpha receptor antagonist, R-7050, improves neurological outcomes following intracerebral hemorrhage in mice. Neurosci Lett. 2013 May 10;542:92-6. doi: 10.1016/j.neulet.2013.02.051. Epub 2013 Mar 7. PMID: 23499961; PMCID: PMC3744337.
[2]. Gururaja TL, Yung S, et,al. A class of small molecules that inhibit TNFalpha-induced survival and death pathways via prevention of interactions between TNFalphaRI, TRADD, and RIP1. Chem Biol. 2007 Oct;14(10):1105-18. doi: 10.1016/j.chembiol.2007.08.012. PMID: 17961823.
[3]. Pal A, Tapadar P, Pal R. Exploring the Molecular Mechanism of Cinnamic Acid-Mediated Cytotoxicity in Triple Negative MDA-MB-231 Breast Cancer Cells. Anticancer Agents Med Chem. 2021;21(9):1141-1150. doi: 10.2174/1871520620666200807222248. PMID: 32767960.
[4]. Zhong B, Ma S, et,al. Ablation of TRPV1 Elevates Nocturnal Blood Pressure in Western Diet-fed Mice. Curr Hypertens Rev. 2019;15(2):144-153. doi: 10.2174/1573402114666181031141840. PMID: 30381083; PMCID: PMC6635649.
[5]. Liu J, Que K, et,al. Tumor Necrosis Factor-α Regulates the TRPA1 Expression in Human Odontoblast-Like Cells. J Pain Res. 2020 Jul 6;13:1655-1664. doi: 10.2147/JPR.S255288. PMID: 32753941; PMCID: PMC7352379.
[6]. Wen Y, Lu X, et,al. KLF4 in Macrophages Attenuates TNFα-Mediated Kidney Injury and Fibrosis. J Am Soc Nephrol. 2019 Oct;30(10):1925-1938. doi: 10.1681/ASN.2019020111. Epub 2019 Jul 23. PMID: 31337692; PMCID: PMC6779357.
[7]. Ayo TE, Adhikari P, et,al. TNF Production in Activated RBL-2H3 Cells Requires Munc13-4. Inflammation. 2020 Apr;43(2):744-751. doi: 10.1007/s10753-019-01161-4. PMID: 31897916; PMCID: PMC7176528.
R-7050 是一种新型细胞渗透性三唑并喹喔啉化合物，可选择性抑制 TNF-α；诱导细胞信号传导[2]。
在 RBL-2H3 细胞中，当自噬也被 Baf-A1 抑制时，R-7050 显着降低 TNF 增殖[7].CA 降低 MDA-MB-231 乳腺癌细胞和致瘤性 HEK 293 细胞的细胞活力，但不影响正常的 NIH3T3 成纤维细胞。 TNFA、TNF 受体 1 (TNFR1) 和裂解的 caspase-8/-3 促凋亡蛋白的丰度随着 CA 处理而增加。小分子抑制剂 R-7050 阻断 TNFA-TNFR1 信号通路，在蛋白质水平上降低了裂解的 caspase-8 和 caspase-3 的表达[3]。 TRPA1通道在HODs的细胞体和过程中呈阳性表达。高浓度 TNF-α 可显着上调 TRPA1 通道的表达，并可被 R-7050[5] 抑制。
单次施用 R-7050 可减少 ICH 后的神经血管损伤。与直接结合和中和 TNF-α 的生物学方法相反；活性，R-7050 选择性抑制 TNFR 与细胞内衔接分子（如 TRADD 和 RIP1）的结合，限制受体内化并阻止 TNF-α 后的后续细胞反应；绑定[1]。辣椒素是一种选择性 TRPV1 激动剂，可增加 WT 而非 TRPV1-/- 小鼠的同侧传入肾神经活动。 WD 摄入增加了瘦素、IL-6 和 TNF-α；在脂肪组织中，和 TNF-α；拮抗剂 III，R-7050，减少 TRPV1-/-WD 中的瘦素。 TRPV1-/--WD组的尿白蛋白水平高于WT-WD[4]。在肾毒性血清肾炎期间使用 TNF 受体 1 抑制剂 R-7050 治疗可减少野生型小鼠和 KLF4 缺陷型巨噬细胞小鼠的损伤、纤维化和坏死性凋亡，并消除两组在这些参数上的差异[ 6].
| Cas No. | 303997-35-5 | SDF | |
| 别名 | TNF-α Antagonist III | ||
| 化学名 | 8-chloro-4-(phenylthio)-1-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]quinoxaline | ||
| Canonical SMILES | ClC1=CC(N2C3=NN=C2C(F)(F)F)=C(C=C1)N=C3SC4=CC=CC=C4 | ||
| 分子式 | C16H8ClF3N4S | 分子量 | 380.8 |
| 溶解度 | DMSO (warmed with 50ºC water bath) 9 mg/mL (23.64 mM) ;DMF: 10 mg/ml | 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。 | ||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.6261 mL | 13.1303 mL | 26.2605 mL |
| 5 mM | 0.5252 mL | 2.6261 mL | 5.2521 mL |
| 10 mM | 0.2626 mL | 1.313 mL | 2.6261 mL |
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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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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，
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2.
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KLF4 in Macrophages Attenuates TNF ¦Á-Mediated Kidney Injury and Fibrosis
J Am Soc Nephrol2019 Oct;30(10):1925-1938.PMID: 31337692DOI: 10.1681/ASN.2019020111
Background: Polarized macrophage populations can orchestrate both inflammation of the kidney and tissue repair during CKD. Proinflammatory M1 macrophages initiate kidney injury, but mechanisms through which persistent M1-dependent kidney damage culminates in fibrosis require elucidation. Kr¨¹ppel-like factor 4 (KLF4), a zinc-finger transcription factor that suppresses inflammatory signals, is an essential regulator of macrophage polarization in adipose tissues, but the effect of myeloid KLF4 on CKD progression is unknown. Methods: We used conditional mutant mice lacking KLF4 or TNF¦Á (KLF4's downstream effector) selectively in myeloid cells to investigate macrophage KLF4's role in modulating CKD progression in two models of CKD that feature robust macrophage accumulation, nephrotoxic serum nephritis, and unilateral ureteral obstruction. Results: In these murine CKD models, KLF4 deficiency in macrophages infiltrating the kidney augmented their M1 polarization and exacerbated glomerular matrix deposition and tubular epithelial damage. During the induced injury in these models, macrophage-specific KLF4 deletion also exacerbated kidney fibrosis, with increased levels of collagen 1 and ¦Á-smooth muscle actin in the injured kidney. CD11b+Ly6Chi myeloid cells isolated from injured kidneys expressed higher levels of TNF¦Á mRNA versus wild-type controls. In turn, mice bearing macrophage-specific deletion of TNF¦Á exhibited decreased glomerular and tubular damage and attenuated kidney fibrosis in the models. Moreover, treatment with the TNF receptor-1 inhibitor R-7050 during nephrotoxic serum nephritis reduced damage, fibrosis, and necroptosis in wild-type mice and mice with KLF4-deficient macrophages, and abrogated the differences between the two groups in these parameters. Conclusions: These data indicate that macrophage KLF4 ameliorates CKD by mitigating TNF-dependent injury and fibrosis.
TNF-alpha receptor antagonist, R-7050, improves neurological outcomes following intracerebral hemorrhage in mice
Neurosci Lett2013 May 10;542:92-6.PMID: 23499961DOI: 10.1016/j.neulet.2013.02.051
Intracerebral hemorrhage (ICH), the most common form of hemorrhagic stroke, exhibits the highest acute mortality and the worst long-term prognosis of all stroke subtypes. Unfortunately, treatment options for ICH are lacking due in part to a lack of feasible therapeutic targets. Inflammatory activation is associated with neurological deficits in pre-clinical ICH models and with patient deterioration after clinical ICH. In the present study, we tested the hypothesis that R-7050, a novel cell permeable triazoloquinoxaline inhibitor of the tumor necrosis factor receptor (TNFR) complex, attenuates neurovascular injury after ICH in mice. Up to 2h post-injury administration of R-7050 significantly reduced blood-brain barrier opening and attenuated edema development at 24h post-ICH. Neurological outcomes were also improved over the first 3 days after injury. In contrast, R-7050 did not reduce hematoma volume, suggesting the beneficial effects of TNFR inhibition were downstream of clot formation/resolution. These data suggest a potential clinical utility for TNFR antagonists as an adjunct therapy to reduce neurological injury and improve patient outcomes after ICH.
TNF-¦Á Receptor Inhibitor Alleviates Metabolic and Inflammatory Changes in a Rat Model of Ischemic Stroke
Antioxidants (Basel)2021 May 26;10(6):851.PMID: 34073455DOI: 10.3390/antiox10060851
Hyperglycemia and inflammation, with their augmented interplay, are involved in cases of stroke with poor outcomes. Interrupting this vicious cycle thus has the potential to prevent stroke disease progression. Tumor necrosis factor-¦Á (TNF-¦Á) is an emerging molecule, which has inflammatory and metabolic roles. Studies have shown that TNF-¦Á receptor inhibitor R-7050 possesses neuroprotective, antihyperglycemic, and anti-inflammatory effects. Using a rat model of permanent cerebral ischemia, pretreatment with R-7050 offered protection against poststroke neurological deficits, brain infarction, edema, oxidative stress, and caspase 3 activation. In the injured cortical tissues, R-7050 reversed the activation of TNF receptor-I (TNFRI), NF-¦ʂ, and interleukin-6 (IL-6), as well as the reduction of zonula occludens-1 (ZO-1). In the in vitro study on bEnd.3 endothelial cells, R-7050 reduced the decline of ZO-1 levels after TNF-¦Á-exposure. R-7050 also reduced the metabolic alterations occurring after ischemic stroke, such as hyperglycemia and increased plasma corticosterone, free fatty acids, C reactive protein, and fibroblast growth factor-15 concentrations. In the gastrocnemius muscles of rats with stroke, R-7050 improved activated TNFRI/NF-¦ʂ, oxidative stress, and IL-6 pathways, as well as impaired insulin signaling. Overall, our findings highlight a feasible way to combat stroke disease based on an anti-TNF therapy that involves anti-inflammatory and metabolic mechanisms.
Microglia-derived TNF-¦Á mediates M¨¹ller cell activation by activating the TNFR1-NF-¦ʂ pathway
Exp Eye Res2022 Jan;214:108852.PMID: 34801535DOI: 10.1016/j.exer.2021.108852
Microglia and its interaction with M¨¹ller cells are responsible to retinal surveillance during retinal neurodegeneration, however, the role and mechanism of microglia-derived tumor necrosis factor (TNF)-¦Á in the activation of retinal M¨¹ller cells have not been fully elucidated. In the present study, primary microglia and M¨¹ller cells were isolated from newborn Sprague-Dawley (SD) rats with purities of 88.2 ¡À 6.2% and 92.2 ¡À 2.2%, respectively. By performing immunofluorescence and Western blot analysis, we found that TNF receptor (TNFR)-1 and TNFR2 were expressed in M¨¹ller cells. After co-cultured with microglia-conditioned medium (MCM), the elevated mRNA levels of glial fibrillary acidic protein (GFAP), proinflammatory factors (TNF-¦Á, IL-1¦¬ CXCL-1, CSF-1, NOS2, COX2) and decreased CNTF mRNA levels were found in M¨¹ller cells. However, pretreatment with R-7050 (a TNF-¦Á receptor inhibitor) or anti-TNFR1 significantly abrogated the changes. Simultaneously, pretreatment with anti-TNFR2 slightly inhibited the expression of GFAP in MCM-incubated M¨¹ller cells. Meanwhile, anti-TNFR1 treatment reversed the increased expression of CSF-1 and IL-1¦ induced by TNF-¦Á. Compared to the control groups, the phosphorylation of NF-¦ʂ P65, MAPK P38 and ERK1/2 in TNF-¦Á-treated M¨¹ller cells was significantly increased. Nevertheless, pretreatment with anti-TNFR1 inhibited the phosphorylation of NF-¦ʂ P65 and MAPK p38, especially NF-¦ʂ P65. Additionally, pretreatment with Bay117082 (an NF-¦ʂ inhibitor) also significantly inhibited NF-¦ʂ P65 phosphorylation and GFAP expression. Moreover, anti-TNFR1 and Bay117082 treatment reduced NF-¦ʂ P65 phosphorylation of M¨¹ller cells induced by MCM. These results suggested that microglia-derived TNF-¦Á served as a vital role in regulating M¨¹ller cells activation during retinal neurodegeneration.
TNF Production in Activated RBL-2H3 Cells Requires Munc13-4
Inflammation2020 Apr;43(2):744-751.PMID: 31897916DOI: 10.1007/s10753-019-01161-4
Mast cell activation triggers intricate signaling pathways that promote the expression and/or release of a wide range of mediators including tumor necrosis factor (TNF; also known as TNF¦Á). In this study, we investigated the connection between TNF secretion and TNF production, exploiting RBL-2H3 cells (a tumor analog of mucosal mast cells) that are depleted of Munc13-4, a crucial component of the mast cell exocytic machinery. We showed that antigen/IgE elicited robust TNF production in RBL-2H3 cells, but not in Munc13-4 knockout cells. The production defect was corrected when Munc13-4 was reintroduced into the knockout cell line, suggesting that the phenotype was not caused by any secondary effect derived from the knockout approach. Furthermore, pre-incubation of RBL-2H3 cells with R-7050, an antagonist of TNF receptor-dependent signaling, was shown to block TNF production without inhibiting TNF release. These observations provide fresh evidence for a robust feed-back loop to boost TNF production in activated mast cells.