| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
IRE1alpha kinase-IN-2 specifically targets the IRE1alpha serine/threonine kinase. It functions as an ATP-competitive inhibitor, binding to the kinase active site and blocking IRE1alpha autophosphorylation (IC50 = 3.12 microM). This, in turn, inhibits its downstream endoribonuclease (RNase) activity, preventing the splicing of XBP1 mRNA (EC50 = 0.82 microM), a critical step in the IRE1alpha/XBP1s arm of the UPR signaling pathway.
|
|---|---|
| ln Vitro |
Compound 3, IRE1α kinase-IN-2, prevents XBP1 mRNA splicing even when the ER is under stress[1].
In vitro, IRE1alpha kinase-IN-2 potently inhibits XBP1 mRNA splicing, even under conditions of ER stress induced by agents like tunicamycin. It impairs IRE1alpha kinase autophosphorylation, which is a prerequisite for its RNase activity. By blocking the IRE1alpha-XBP1s axis, it reduces the cell's adaptive capacity to ER stress, which can lead to apoptosis in cells that rely on this pathway for survival, such as multiple myeloma cells. |
| ln Vivo |
Specific in vivo efficacy data for IRE1alpha kinase-IN-2 is not detailed in the provided literature. However, its potent mechanism of action suggests it could be used in mouse xenograft models of cancers that are addicted to the IRE1alpha/XBP1s pathway. Oral administration of a similar IRE1alpha inhibitor has been shown to reduce tumor growth and decrease XBP1s splicing in tumor tissue, validating its potential for in vivo use.
|
| Enzyme Assay |
For a cell-free system, a radiometric or luminescent kinase assay is used. Purified IRE1alpha kinase domain is incubated with ATP and a peptide substrate. The compound is added to the reaction mixture. The level of peptide phosphorylation is measured, and the IC50 for inhibition of autophosphorylation (3.12 microM) is calculated. The RNase activity can be measured in vitro using purified IRE1alpha and a synthetic RNA stem-loop substrate representing XBP1.
|
| Cell Assay |
In cellular assays, HeLa or other cell lines are treated with an ER stress inducer (e.g., tunicamycin or thapsigargin) in the presence or absence of IRE1alpha kinase-IN-2. The level of spliced XBP1 (XBP1s) is measured by RT-qPCR (using primers that amplify the spliced isoform) or by a luciferase reporter assay containing the XBP1s splicing site. A reduction in the EC50 (0.82 microM) confirms the compound's efficacy.
|
| Animal Protocol |
For in vivo studies, mice bearing human tumor xenografts (e.g., multiple myeloma) are treated with IRE1alpha kinase-IN-2. The compound can be formulated for intraperitoneal (IP) injection using a vehicle such as 5% DMSO + 40% PEG300 + 5% Tween-80 + 50% ddH2O. Tumor volume is measured with calipers, and at the end of the study, tumor tissue is harvested to quantify XBP1s levels by RT-qPCR as a pharmacodynamic marker.
|
| ADME/Pharmacokinetics |
Specific ADME data for IRE1alpha kinase-IN-2 is not provided. Its molecular weight (278.31) suggests it falls within the "Lipinski's Rule of Five" for drug-likeness. It is soluble in DMSO at 11.11 mg/mL, allowing for stock solution preparation. For in vivo use, a formulation to enhance solubility (e.g., in 10% DMSO + 90% saline) might be required. It is stable as a powder at -20degC for up to three years.
|
| Toxicity/Toxicokinetics |
No specific toxicology data for IRE1alpha kinase-IN-2 is presented. As an inhibitor of a key stress response pathway, its on-target toxicity could involve the disruption of normal ER homeostasis in tissues that experience high secretory demand, such as the pancreas, liver, and plasma cells. The safety profile of IRE1alpha inhibition is an active area of investigation in drug discovery.
|
| References | |
| Additional Infomation |
IRE1alpha is a dual-function enzyme possessing both kinase and RNase activities, making it a unique and complex drug target. IRE1alpha kinase-IN-2 is a tool compound used to dissect the relative contributions of IRE1alpha's kinase and RNase domains. The inhibition of XBP1 splicing is a key biomarker for UPR activity. This compound is a preclinical research tool and is not approved for clinical use.
|
| Molecular Formula |
C27H23F3N6O
|
|---|---|
| Molecular Weight |
504.51
|
| Exact Mass |
504.188
|
| CAS # |
1414938-21-8
|
| PubChem CID |
60196244
|
| Appearance |
White to off-white solid powder
|
| Density |
1.4±0.1 g/cm3
|
| Index of Refraction |
1.660
|
| LogP |
5.92
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
37
|
| Complexity |
797
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CC(C)C1=NC(=C2N1C=CN=C2N)C3=CC=C(C4=CC=CC=C43)NC(=O)NC5=CC=CC(=C5)C(F)(F)F
|
| InChi Key |
MEJKZYOOTMLMBA-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C27H23F3N6O/c1-15(2)25-35-22(23-24(31)32-12-13-36(23)25)20-10-11-21(19-9-4-3-8-18(19)20)34-26(37)33-17-7-5-6-16(14-17)27(28,29)30/h3-15H,1-2H3,(H2,31,32)(H2,33,34,37)
|
| Chemical Name |
1-[4-(8-amino-3-propan-2-ylimidazo[1,5-a]pyrazin-1-yl)naphthalen-1-yl]-3-[3-(trifluoromethyl)phenyl]urea
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO: 100 mg/mL (198.21 mM)
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9821 mL | 9.9106 mL | 19.8212 mL | |
| 5 mM | 0.3964 mL | 1.9821 mL | 3.9642 mL | |
| 10 mM | 0.1982 mL | 0.9911 mL | 1.9821 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
