ISO-1
(Synonyms: 4,5-二氢-3-(4-羟基苯基)-5-异恶唑乙酸甲酯,MIF Antagonist) 目录号 : GC17108
ISO-1以剂量依赖的方式抑制MIF互变异构酶活性，IC50约为7µM。
Cas No.:478336-92-4
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Kinase experiment [1]: | |
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Preparation Method |
Activity was determined at room temperature by adding dopachrome methyl ester (0.3 ml) to a cuvette containing 50 nm MIF in 50 mm potassium phosphate buffer, pH 6, 0.5 mm EDTA and measuring the decrease in absorbance from 2 to 20 s at 475 nm spectrophotometrically. The inhibitors were dissolved in Me2SO at various concentrations and added to the cuvette with the MIF prior to the addition of the dopachrome. |
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Reaction Conditions |
1uM-1000uM ISO-1 |
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Applications |
ISO-1 inhibited MIF tautomerase activity in a dose-dependent manner with an IC50 of about 7 µM, but the non-hydroxylated phenyl analog (compound 2) was 10-15 times less potent |
| Cell experiment [1]: | |
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Cell lines |
RAW 264.7 macrophages |
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Preparation Method |
Cells were subsequently transfected with FuGENE 6 and pcDNA/GS empty vector or FuGENE 6 and pcDNA/GS/MIF in the absence or presence of vehicle (Me2SO) or inhibitors. After 16 h cell supernatants were collected and14C-labeled arachidonic acid was assessed by scintillation counting. |
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Reaction Conditions |
1-100uM ISO-1 for 16 h |
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Applications |
Treatment of the transfected cells with ISO-1 inhibited the release of arachidonic acid in a dose-dependent manner, the MIF inhibitory activities of ISO-1 and its derivatives were not the result of cell toxicity. |
| Animal experiment [2]: | |
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Animal models |
Male Balb/C mice, 8 weeks old |
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Preparation Method |
Endotoxemia was induced by injection of a sublethal dose of LPS. Mice were treated with various concentrations of ISO-1 (3.5-35 mg/kg; intraperitoneally) or vehicle (aqueous 5% dimethyl sulfoxide) 30 min before and 6 h after LPS infusion and then twice daily for 3 days. Animals were monitored for survival for 2 weeks. |
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Dosage form |
ISO-1 (3.5-35 mg/kg; intraperitoneally) for 3 days |
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Applications |
ISO-1 inhibits tumor necrosis factor release from macrophages isolated from LPStreated wild type mice but has no effect on cytokine release from MIFdeficient macrophages. |
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References: [1]. Lubetsky JB, Dios A, et,al.The tautomerase active site of macrophage migration inhibitory factor is a potential target for discovery of novel anti-inflammatory agents. J Biol Chem. 2002 Jul 12;277(28):24976-82. doi: 10.1074/jbc.M203220200. Epub 2002 May 7. PMID: 11997397. |
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ISO-1 inhibited MIF tautomerase activity in a dose-dependent manner with an IC50 of about 7 µM[1].
Treatment of the transfected cells with ISO-1 inhibited the release of arachidonic acid in a dose-dependent manner, the MIF inhibitory activities of ISO-1 and its derivatives were not the result of cell toxicity[1]. In monocytes, ISO-1 caused marked suppression of TLR4-induced proinflammatory cytokine production, and to a lesser extent suppression of TLR2-induced responses[6].
ISO-1 inhibits tumor necrosis factor release from macrophages isolated from LPStreated wild type mice but has no effect on cytokine release from MIFdeficient macrophages[2]. ISO-1 exerts anti-cancer effects on PANC-1 cell proliferation, migration and invasion in vitro, and inhibited PANC-1 cell-induced tumour growth in xenograft mice in vivo[3]. ISO-1 treatment alleviated pathological damage in pancreatic and renal tissues, and reduced the serum levels of amylase, lipase, creatinine, uric acid, IL-6 and TNF-α. ISO-1 also reduced protein expression of NLRP3, ASC, caspase-1 and IL-1β, mRNA expression of MIF, IL-6, TNF-α, IL-1β and IL-18, and the infiltration of MPO-positive neutrophils in kidney tissue[4]. ISO-1 may protect the IBD cells, reduce pathological injuries, and reduce the inflammatory response in SAP rats. Its mechanisms may be via inhibiting the expression of MIF and then blocking the activation of p38-MAPK and NF-κB signaling pathway[5]. ISO-1 remarkably improved the histological findings of APAP-induced liver injury in mice. The increases in serum levels of alanine aminotransferase (ALT), and macrophage inflammatory protein-2 (MIP-2) by APAP were inhibited by ISO-1[7].
References:
[1]. Lubetsky JB, Dios A, et,al.The tautomerase active site of macrophage migration inhibitory factor is a potential target for discovery of novel anti-inflammatory agents. J Biol Chem. 2002 Jul 12;277(28):24976-82. doi: 10.1074/jbc.M203220200. Epub 2002 May 7. PMID: 11997397.
[2]. Al-Abed Y, Dabideen D, et,al. ISO-1 binding to the tautomerase active site of MIF inhibits its pro-inflammatory activity and increases survival in severe sepsis. J Biol Chem. 2005 Nov 4;280(44):36541-4. doi: 10.1074/jbc.C500243200. Epub 2005 Aug 22. PMID: 16115897.
[3]. Cheng B, Wang Q, et,al.MIF inhibitor, ISO-1, attenuates human pancreatic cancer cell proliferation, migration and invasion in vitro, and suppresses xenograft tumour growth in vivo. Sci Rep. 2020 Apr 21;10(1):6741. doi: 10.1038/s41598-020-63778-y. PMID: 32317702; PMCID: PMC7174354.
[4]. Liu Y, Liu Y, et,al. MIF inhibitor ISO-1 alleviates severe acute pancreatitis-associated acute kidney injury by suppressing the NLRP3 inflammasome signaling pathway. Int Immunopharmacol. 2021 Jul;96:107555. doi: 10.1016/j.intimp.2021.107555. Epub 2021 Apr 3. PMID: 33823428.
[5]. Wang B, Zhao K, et,al. Protective Mechanism of MIF Inhibitor ISO-1 on Intrahepatic Bile Duct Cells in Rats with Severe Acute Pancreatitis. Dig Dis Sci. 2021 Oct;66(10):3415-3426. doi: 10.1007/s10620-020-06674-9. Epub 2020 Oct 29. PMID: 33123939.
[6]. West PW, Parker LC, et,al. Differential and cell-type specific regulation of responses to Toll-like receptor agonists by ISO-1. Immunology. 2008 Sep;125(1):101-10. doi: 10.1111/j.1365-2567.2008.02825.x. Epub 2008 Mar 18. PMID: 18355244; PMCID: PMC2526264.
[7]. Ohkawara T, Okubo N, et,al. Protective effect of ISO-1 with inhibition of RIPK3 up-regulation and neutrophilic accumulation on acetaminophen-induced liver injury in mice. Toxicol Lett. 2021 Mar 15;339:51-59. doi: 10.1016/j.toxlet.2020.12.015. Epub 2020 Dec 25. PMID: 33370591.
ISO-1以剂量依赖的方式抑制MIF互变异构酶活性，IC50约为7µM[1]。
转染细胞的ISO-处理1以剂量依赖方式抑制花生四烯酸的释放，ISO-1及其衍生物的MIF抑制活性不是细胞毒性的结果[1]。在单核细胞中，ISO-1 显着抑制 TLR4 诱导的促炎细胞因子产生，并在较小程度上抑制 TLR2 诱导的反应[6]。
ISO-1 抑制从 LPS 处理的野生型小鼠分离出的巨噬细胞释放肿瘤坏死因子，但对 MIF 缺陷型巨噬细胞释放细胞因子没有影响[2]。 ISO-1 在体外对 PANC-1 细胞增殖、迁移和侵袭发挥抗癌作用，在体内抑制 PANC-1 细胞诱导的异种移植小鼠肿瘤生长[3]。 ISO-1 治疗减轻了胰腺和肾脏组织的病理损伤，并降低了淀粉酶、脂肪酶、肌酐、尿酸、IL-6 和 TNF-α 的血清水平；。 ISO-1 还降低了 NLRP3、ASC、caspase-1 和 IL-1β 的蛋白表达；MIF、IL-6、TNF-α 的 mRNA 表达；IL-1β；和 IL-18，以及肾组织中 MPO 阳性中性粒细胞的浸润[4]。 ISO-1 可能保护 IBD 细胞，减少病理损伤，并减少 SAP 大鼠的炎症反应。其机制可能是通过抑制MIF的表达进而阻断p38-MAPK和NF-κB信号通路的激活[5]。 ISO-1 显着改善了 APAP 诱导的小鼠肝损伤的组织学发现。 APAP对血清谷丙转氨酶（ALT）和巨噬细胞炎症蛋白2（MIP-2）水平的升高可被ISO-1[7]抑制。
| Cas No. | 478336-92-4 | SDF | |
| 别名 | 4,5-二氢-3-(4-羟基苯基)-5-异恶唑乙酸甲酯,MIF Antagonist | ||
| 分子式 | C12H13NO4 | 分子量 | 235.24 |
| 溶解度 | DMF: 20 mg/ml,DMF:PBS(pH 7.2)(1:1): 0.5 mg/ml,DMSO: 14 mg/ml,Ethanol: 5 mg/ml | 储存条件 | Store at -20°C |
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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 | 4.251 mL | 21.2549 mL | 42.5098 mL |
| 5 mM | 0.8502 mL | 4.251 mL | 8.502 mL |
| 10 mM | 0.4251 mL | 2.1255 mL | 4.251 mL |
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Yeast iso-1-cytochrome c: genetic analysis of structural requirements
We describe the use of classical and molecular genetic techniques to investigate the folding, stability, and enzymatic requirements of iso-1-cytochrome c from the yeast Saccharomyces cerevisiae. Interpretation of the defects associated with an extensive series of altered forms of iso-1-cytochrome c was facilitated by the recently resolved three dimensional structure of iso-1-cytochrome c [(1987) J. Mol. Biol. 199, 295-314], and by comparison with the phylogenetic series of eukaryotic cytochromes c. Residue replacements that abolish iso-1-cytochrome c function appear to do so by affecting either heme attachment or protein stability; no replacements that abolish electron transfer function without affecting protein structure were uncovered. Most nonfunctional forms retained at least partial covalent attachment to the heme moiety; heme attachment was abolished only by replacements of Cys19 and Cys22, which are required for thioether linkage, and His23, a heme ligand. Replacements were uncovered that retain function at varying levels, including replacements at evolutionarily conserved positions, some of which were structurally and functionally indistinguishable from wild type iso-1-cytochrome c.
Breast Cancer 18F-ISO-1 Uptake as a Marker of Proliferation Status
The σ2 receptor is a potential in vivo target for measuring proliferative status in cancer. The feasibility of using N-(4-(6,7-dimethoxy-3,4-dihydroisoquinolin-2(1H)-yl)butyl)-2-(2-18F-fluoroethoxy)-5-methylbenzamide (18F-ISO-1) to image solid tumors in lymphoma, breast cancer, and head and neck cancer has been previously established. Here, we report the results of the first dedicated clinical trial of 18F-ISO-1 in women with primary breast cancer. Our study objective was to determine whether 18F-ISO-1 PET could provide an in vivo measure of tumor proliferative status, and we hypothesized that uptake would correlate with a tissue-based assay of proliferation, namely Ki-67 expression. Methods: Twenty-eight women with 29 primary invasive breast cancers were prospectively enrolled in a clinical trial (NCT02284919) between March 2015 and January 2017. Each received an injection of 278-527 MBq of 18F-ISO-1 and then underwent PET/CT imaging of the breasts 50-55 min later. In vivo uptake of 18F-ISO-1 was quantitated by SUVmax and distribution volume ratios and was compared with ex vivo immunohistochemistry for Ki-67. Wilcoxon rank-sum tests assessed uptake differences across Ki-67 thresholds, and Spearman correlation tested associations between uptake and Ki-67. Results: Tumor SUVmax (median, 2.0 g/mL; range, 1.3-3.3 g/mL), partial-volume-corrected SUVmax, and SUV ratios were tested against Ki-67. Tumors stratified into the high-Ki-67 (≥20%) group had SUVmax greater than the low-Ki-67 (<20%) group (P = 0.02). SUVmax exhibited a positive correlation with Ki-67 across all breast cancer subtypes (ρ = 0.46, P = 0.01, n = 29). Partial-volume-corrected SUVmax was positively correlated with Ki-67 for invasive ductal carcinoma (ρ = 0.51, P = 0.02, n = 21). Tumor-to-normal-tissue ratios and tumor distribution volume ratio did not correlate with Ki-67 (P > 0.05). Conclusion:18F-ISO-1 uptake in breast cancer modestly correlates with an in vitro assay of proliferation.
MIF inhibitor ISO-1 alleviates severe acute pancreatitis-associated acute kidney injury by suppressing the NLRP3 inflammasome signaling pathway
Background: Acute kidney injury (AKI) is an important complication of severe acute pancreatitis (SAP) with a poor prognosis. The methyl ester of (S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid (ISO-1), an inhibitor of macrophage migration inhibitory factor (MIF), has protective effects against many diseases. Our previous study confirmed MIF inhibition alleviated SAP. Here, we explored the effects of ISO-1 in an experimental mouse model of SAP-associated AKI induced by l-arginine.
Methods: Mice were randomly divided into four treatment groups (n = 6 each): control (CON), SAP, SAP + ISO-1, and ISO-1. Histopathologic examination was used to observe damage in pancreatic and renal tissues. Biochemical and enzyme-linked immunosorbent assays (ELISA) kits were used to measure the serologic indicators amylase, lipase, creatinine, uric acid, interleukin (IL)-6, and tumor necrosis factor (TNF)-α. Immunohistochemistry was used to detect protein expression of NLRP3, ASC and caspase-1, and the infiltration of myeloperoxidase (MPO)-positive neutrophils in kidney tissue. Western blotting was used to detect NLRP3, ASC and caspase-1 and IL-1β protein expression, and real-time PCR was used to measure MIF, IL-6, TNF-α, IL-1β and IL-18 mRNA levels in kidney tissue.
Results: ISO-1 treatment alleviated pathological damage in pancreatic and renal tissues, and reduced the serum levels of amylase, lipase, creatinine, uric acid, IL-6 and TNF-α. ISO-1 also reduced protein expression of NLRP3, ASC, caspase-1 and IL-1β, mRNA expression of MIF, IL-6, TNF-α, IL-1β and IL-18, and the infiltration of MPO-positive neutrophils in kidney tissue.
Conclusion: ISO-1 has a protective effect against experimental SAP-associated AKI. And the mechanism may be associated with ISO-1 inhibiting NLRP3 inflammasome signaling pathway.
Protective effect of ISO-1 with inhibition of RIPK3 up-regulation and neutrophilic accumulation on acetaminophen-induced liver injury in mice
Overdose use of acetaminophen (APAP) often occurs a severe liver injury, and its liver injury is lethal in some cases. Macrophage migration inhibitory factor (MIF) is expressed in a variety of cells and has multifunctional roles. However, the role of MIF in APAP-induced liver injury has not been fully investigated. In this study, we investigated whether treatment with (S,R)-3-(4-hydroxyphenil)-4,5-dihydro-5-isoxazole acetic acid methyl ester (ISO-1), a MIF inhibitor, protected mice from acute APAP-induced liver injury. Acute liver injury was induced by injection of APAP (300 mg/kg body weight). Mice were treated with a single injection of ISO-1(15 mg/kg body weight) 1 h (h) before APAP administration. Histological, biochemical and molecular analyses were performed in liver of mice 12 h after APAP administration. ISO-1 remarkably improved the histological findings of APAP-induced liver injury in mice. The increases in serum levels of alanine aminotransferase (ALT), and macrophage inflammatory protein-2 (MIP-2) by APAP were inhibited by ISO-1. In addition, ISO-1 reduced the increased number of the myeloperoxidase-staining cells and that of TUNEL-positive staining cells in the liver of mice with APAP-induced liver injury. Up-regulation of hepatic receptor interacting protein kinase (RIPK)3 and heat shock protein70 by APAP was suppressed in the liver of mice given ISO-1. These results provide the additional evidence that inhibition of MIF activity may be clinically effective for treatment of acute APAP-induced liver injury.
MIF as a disease target: ISO-1 as a proof-of-concept therapeutic
Macrophage migration inhibitory factor (MIF) is a pleiotropic proinflammatory cytokine that has been implicated as playing a causative role in many disease states, including sepsis, pneumonia, diabetes, rheumatoid arthritis, inflammatory bowel disease, psoriasis and cancer. To inhibit the enzymatic and biologic activities of MIF, we and others have developed small-molecule MIF inhibitors. Most MIF inhibitors bind within the hydrophobic pocket that contains highly conserved amino acids known to be essential for MIF's proinflammatory activity. The best characterized of these small-molecule MIF inhibitors, (S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester (ISO-1) has been validated in scores of laboratories worldwide. Like neutralizing anti-MIF antibodies, ISO-1 significantly improves survival and reduces disease progression and/or severity in multiple murine models where MIF is implicated. This MIF inhibitor, its derivatives and other MIF-targeted compounds show great promise for future testing in disease states where increased MIF activity has been discovered.