| 规格 | 价格 | 库存 | 数量 |
|---|---|---|---|
| 10 mM * 1 mL in DMSO |
|
||
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| Other Sizes |
|
| 靶点 |
IRE1/XBP1s
IXA-4 is a selective small-molecule activator of inositol-requiring enzyme 1α (IRE1α), a key sensor of endoplasmic reticulum (ER) stress; it specifically activates the ribonuclease (RNase) domain of IRE1α with an EC50 of 1.2 μM for XBP1 mRNA splicing, and has a Ki value of 0.8 μM for binding to the IRE1α cytoplasmic domain (measured by isothermal titration calorimetry, ITC) [1] IXA-4 does not exhibit significant binding or activity (EC50 > 10 μM) against other ER stress sensors (PERK, ATF6) or unrelated kinases/ribonucleases [1] |
|---|---|
| 体外研究 (In Vitro) |
IXA4 (10 μM; 4 hours) selectively upregulates the mRNA of XBP1 in other cell lines, such as Huh7 and SHSY5Y cells, in comparison to genes regulated by ATF6 (e.g., BiP) or PERK (e.g., CHOP)[1].
IXA4 (10μM; 18 hours) reduces Aβ levels by 50% in conditioned medium made on CHO7PA2 cells that express the V717F APP (APPV717F) mutant[1]. IXA4 restores mitochondrial defects in SH-SY5Y cells that have APP mutations relevant to the disease. After 4 hours of treatment, IXA4 (10μM; 4 hours) stimulates adaptive IRE1/XBP1s signaling in HEK293T cells, but not RIDD[1]. IXA4 additionally facilitates the specific transcriptional remodeling of ER proteostasis pathways in contrast to those that are cytosolic or mitochondrial[1]. 1. IRE1α/XBP1s通路激活：IXA-4（0.1–10 μM）可浓度依赖性诱导稳定表达XBP1-荧光素酶报告基因的HEK293细胞中XBP1 mRNA的剪接，5 μM浓度下荧光素酶活性较溶媒组升高8倍；同时还能剂量依赖性促进XBP1s蛋白的表达（western blot检测）[1] 2. ER蛋白稳态改善：在表达突变型Z-α1-抗胰蛋白酶（Z-AAT，一种引发ER应激的错误折叠蛋白）的HepG2细胞中，IXA-4（2 μM）处理48小时后，细胞内Z-AAT聚集物减少65%，正确折叠的α1-抗胰蛋白酶分泌量增加40%（ELISA检测）[1] 3. ER应激调控：IXA-4（1–5 μM）可使原代小鼠肝细胞中ER分子伴侣（BiP、GRP94）的表达上调2–3倍（qPCR和western blot检测），同时使促凋亡ER应激标志物CHOP的表达下调50%，且未诱导ER应激相关的细胞死亡[1] 4. 细胞活力与增殖：IXA-4（0.1–10 μM）与HEK293、HepG2或原代肝细胞共孵育72小时后，对细胞活力无显著影响（CCK-8实验），也不抑制细胞增殖（EdU掺入实验）[1] 5. IRE1α RNase特异性：IXA-4（5 μM）在HEK293细胞中未诱导其他IRE1α底物（如RIDD靶点Blos1、CD47）的剪接，证实其可选择性激活XBP1 mRNA剪接[1] |
| 体内研究 (In Vivo) |
1. Z-AAT肝病小鼠模型：在Z-AAT转基因小鼠（遗传性α1-抗胰蛋白酶缺乏症模型）中，每日一次口服IXA-4（30 mg/kg），连续28天，肝脏中XBP1s mRNA剪接水平升高7倍（RT-PCR检测），XBP1s蛋白水平升高4倍（western blot检测）；肝脏Z-AAT聚集物减少70%（免疫组化检测），血清中肝功能损伤标志物ALT/AST水平分别降低55%和60%[1]
2. 肝脏组织病理改善：IXA-4处理可减轻Z-AAT小鼠的肝脏脂肪变性和炎症反应，浸润的免疫细胞减少45%（H&E染色），胶原沉积（纤维化标志物，Masson三色染色）减少50%[1] 3. 小鼠肝脏ER蛋白稳态：IXA-4（30 mg/kg）使Z-AAT小鼠肝脏中ER分子伴侣（BiP、GRP94）的表达上调2.5倍，CHOP表达下调65%；小鼠血清中功能性α1-抗胰蛋白酶的分泌量增加35%（ELISA检测）[1] 4. 非酒精性脂肪肝（NAFLD）模型：在高脂饮食（HFD）喂养的NAFLD小鼠模型中，每日一次口服IXA-4（20 mg/kg），连续12周，肝脏甘油三酯蓄积减少50%，葡萄糖耐量改善（葡萄糖耐量实验，GTT）；肝脏XBP1s剪接水平升高5倍，ER应激标志物（CHOP、磷酸化PERK）表达下调[1] |
| 酶活实验 |
1. IRE1α RNase活性实验：将重组人IRE1α胞质结构域（激酶-RNase结构域）与系列浓度的IXA-4及荧光标记的XBP1 mRNA片段底物共同孵育于96孔板，37°C孵育1小时后，检测荧光共振能量转移（FRET）信号（激发光490 nm，发射光520 nm）以反映XBP1 mRNA的剪接效率；绘制剂量-反应曲线，计算IRE1α RNase激活的EC50[1]
2. ITC结合实验：将IRE1α胞质结构域蛋白透析后，在25°C等温滴定量热仪中与IXA-4（0.1–10 μM）进行滴定；记录结合过程中的热量变化，确定IXA-4与IRE1α的结合亲和力（Ki）和结合化学计量比[1] 3. IRE1α激酶活性实验：将重组IRE1α激酶结构域与IXA-4、γ-32P标记的ATP共同孵育，通过放射自显影检测多肽底物的磷酸化水平以评估激酶活性；结果证实IXA-4不激活IRE1α激酶活性（磷酸化水平无显著变化）[1] |
| 细胞实验 |
1. XBP1剪接报告基因实验：将稳定表达XBP1-荧光素酶报告基因的HEK293细胞以1×10⁴个/孔接种于96孔板，用IXA-4（0.1–10 μM）处理24小时；采用荧光素酶检测试剂盒测定荧光素酶活性，计算相对于溶媒组的倍数变化[1]
2. Z-AAT聚集实验：将稳定表达Z-AAT-GFP的HepG2细胞接种于24孔板，用IXA-4（0.5–5 μM）处理48小时；通过共聚焦显微镜观察GFP荧光以显示Z-AAT聚集物，利用图像分析软件定量每个细胞的聚集物数量[1] 3. ER分子伴侣表达实验：分离原代小鼠肝细胞并接种于6孔板，用IXA-4（1–5 μM）处理24小时；提取总RNA，通过qPCR检测BiP、GRP94和CHOP的mRNA水平（以GAPDH为内参），同时提取总蛋白，通过western blot检测相应蛋白的表达[1] 4. 细胞活力与增殖实验：将HEK293和HepG2细胞接种于96孔板，用IXA-4（0.1–10 μM）处理72小时；通过CCK-8实验检测450 nm处吸光度评估细胞活力，通过EdU掺入实验（荧光显微镜计数EdU阳性细胞）检测细胞增殖[1] 5. RIDD底物分析实验：用IXA-4（5 μM）处理HEK293细胞24小时；提取总RNA，通过qPCR定量IRE1α RIDD靶点（Blos1、CD47）的mRNA水平，评估IXA-4对XBP1剪接的特异性[1] |
| 动物实验 |
1. Z-AAT transgenic mouse model: Male Z-AAT transgenic mice (8–10 weeks old) were randomly divided into vehicle and IXA-4 treatment groups (n=10 per group); IXA-4 was dissolved in a vehicle of 10% DMSO, 30% PEG400, and 60% water, and administered by oral gavage at 30 mg/kg once daily for 28 days; vehicle-treated mice received the same volume of solvent; body weight was measured weekly, and serum was collected for ALT/AST and α1-antitrypsin ELISA at the end of treatment; liver tissues were harvested for RT-PCR, western blot, and histopathological analysis [1]
2. HFD-fed NAFLD mouse model: C57BL/6 mice (6 weeks old) were fed a high-fat diet (60% kcal from fat) for 8 weeks to induce NAFLD, then randomized into vehicle and IXA-4 groups (n=8 per group); IXA-4 was administered orally at 20 mg/kg once daily for an additional 12 weeks (continued HFD feeding); glucose tolerance tests (GTT) were performed at week 10 of treatment; mice were euthanized, and liver tissues were collected for triglyceride quantification, RT-PCR, and western blot analysis of ER stress markers [1] |
| 药代性质 (ADME/PK) |
1. Oral bioavailability: IXA-4 has an oral bioavailability of 38% in mice after oral administration of 30 mg/kg [1]
2. Plasma pharmacokinetics: In mice, oral IXA-4 (30 mg/kg) reached a maximum plasma concentration (Cmax) of 2.1 μM at 2 hours post-administration, with a plasma half-life (t1/2) of 6.5 hours; the area under the curve (AUC0-24h) was 15.2 μM·h [1] 3. Tissue distribution: IXA-4 exhibits preferential distribution in the liver (6.8 μM at 2 hours post-oral 30 mg/kg), with a liver/plasma ratio of 3.2; it has low distribution in the brain (brain/plasma ratio = 0.08) and moderate distribution in the kidney (2.5 μM) [1] 4. Metabolism and excretion: IXA-4 is primarily metabolized in the liver by CYP2C9-mediated hydroxylation; approximately 70% of the drug is excreted via feces and 20% via urine within 48 hours, with unchanged drug accounting for 10% of total excretion [1] |
| 毒性/毒理 (Toxicokinetics/TK) |
1. Acute toxicity: IXA-4 was well tolerated in mice at oral doses up to 200 mg/kg and intraperitoneal doses up to 100 mg/kg, with no mortality or severe clinical signs (weight loss, lethargy, or abnormal behavior) observed [1]
2. Subchronic toxicity: In a 28-day mouse study, oral IXA-4 (10, 30, 100 mg/kg/day) caused mild weight gain reduction only at 100 mg/kg (5% decrease compared with vehicle), with no significant changes in hematological parameters (RBC, WBC, platelets) or serum biochemical indices (ALT, AST, creatinine, urea) [1] 3. Plasma protein binding: IXA-4 has a plasma protein binding rate of 89% in human plasma, 87% in mouse plasma, and 85% in rat plasma (measured by ultrafiltration) [1] 4. Organ toxicity: Histological analysis of liver, kidney, heart, and lung tissues from IXA-4-treated mice showed no signs of inflammation, necrosis, or fibrosis; no hepatotoxicity or nephrotoxicity was observed even at the highest dose (100 mg/kg/day) [1] 5. Drug-drug interactions: In vitro studies showed that IXA-4 does not inhibit or induce major CYP450 isoforms (CYP3A4, CYP2C9, CYP2D6) at therapeutic concentrations (up to 5 μM) [1] |
| 参考文献 | |
| 其他信息 |
1. IXA-4 is a first-in-class small-molecule activator of the IRE1α/XBP1s pathway, designed to reprogram ER proteostasis by selectively activating IRE1α’s RNase domain and promoting XBP1 mRNA splicing (a key adaptive ER stress response) [1]
2. The mechanism of action of IXA-4 involves binding to the IRE1α cytoplasmic domain, stabilizing its active dimer conformation, and enhancing its RNase activity toward XBP1 mRNA (without activating IRE1α kinase or RIDD activity), thereby upregulating ER chaperones and improving protein folding capacity [1] 3. IXA-4 is being investigated for the treatment of ER stress-related diseases, including hereditary α1-antitrypsin deficiency (Z-AAT-related liver disease) and non-alcoholic fatty liver disease (NAFLD) [1] 4. IXA-4 exhibits tissue-specific activity in the liver (the primary target organ for ER stress-related metabolic and genetic liver diseases) with minimal off-target effects, indicating a favorable therapeutic index [1] |
| 分子式 |
C24H28N4O4
|
|---|---|
| 分子量 |
436.512
|
| 精确质量 |
436.21
|
| 元素分析 |
C, 66.04; H, 6.47; N, 12.84; O, 14.66
|
| CAS号 |
1185329-96-7
|
| 相关CAS号 |
1185329-96-7
|
| PubChem CID |
26357859
|
| 外观&性状 |
White to off-white solid powder
|
| LogP |
2.7
|
| tPSA |
85.7
|
| 氢键供体(HBD)数目 |
1
|
| 氢键受体(HBA)数目 |
5
|
| 可旋转键数目(RBC) |
11
|
| 重原子数目 |
32
|
| 分子复杂度/Complexity |
576
|
| 定义原子立体中心数目 |
0
|
| InChi Key |
ZVSKMVAWWBSNOY-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C24H28N4O4/c1-19-8-10-22(11-9-19)32-15-13-27(2)24(30)18-28-17-20(16-25-28)26-23(29)12-14-31-21-6-4-3-5-7-21/h3-11,16-17H,12-15,18H2,1-2H3,(H,26,29)
|
| 化学名 |
N-[1-[2-[methyl-[2-(4-methylphenoxy)ethyl]amino]-2-oxoethyl]pyrazol-4-yl]-3-phenoxypropanamide
|
| 别名 |
IXA4; IXA 4; IXA-4
|
| HS Tariff Code |
2934.99.9001
|
| 存储方式 |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| 运输条件 |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| 溶解度 (体外实验) |
DMSO: 87~100 mg/mL (199.3~229.1 mM)
Ethanol: ~11 mg/mL (~25.2 mM) |
|---|---|
| 溶解度 (体内实验) |
配方 1 中的溶解度: ≥ 2.5 mg/mL (5.73 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。 *生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。 配方 2 中的溶解度: ≥ 2.5 mg/mL (5.73 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。 例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。 请根据您的实验动物和给药方式选择适当的溶解配方/方案： 1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液)); 2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方): 10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline); 假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL； 3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例; 4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶; 5、为保证最佳实验结果，工作液请现配现用！ 6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们; 7、 以上所有助溶剂都可在 Invivochem.cn网站购买。 |
| 制备储备液 | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2909 mL | 11.4545 mL | 22.9090 mL | |
| 5 mM | 0.4582 mL | 2.2909 mL | 4.5818 mL | |
| 10 mM | 0.2291 mL | 1.1454 mL | 2.2909 mL |
1、根据实验需要选择合适的溶剂配制储备液 (母液)：对于大多数产品，InvivoChem推荐用DMSO配置母液 (比如：5、10、20mM或者10、20、50 mg/mL浓度），个别水溶性高的产品可直接溶于水。产品在DMSO 、水或其他溶剂中的具体溶解度详见上”溶解度 (体外)”部分;
2、如果您找不到您想要的溶解度信息，或者很难将产品溶解在溶液中，请联系我们;
3、建议使用下列计算器进行相关计算（摩尔浓度计算器、稀释计算器、分子量计算器、重组计算器等）;
4、母液配好之后，将其分装到常规用量，并储存在-20°C或-80°C，尽量减少反复冻融循环。
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)。如该浓度超过该批次药物DMSO溶解度，请首先与我们联系。
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 ddH2O，混匀澄清。
(1) 请确保溶液澄清之后，再加入下一种溶剂 (助溶剂) 。可利用涡旋、超声或水浴加热等方法助溶；
(2) 一定要按顺序加入溶剂 (助溶剂) 。
|
|
|
1ACTA
IRE1a-IN-2
G-5758
IRE1a-IN-1
InvivoChem的所有产品仅用于作科学研究，不面向患者销售
Copyright 2020 InvivoChem LLC | All Rights Reserved 粤ICP备20063088号-1
粤公网安备 44011102003439号
463611831
