| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg | |||
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| Targets |
RIPK2 kinase (IC50 = 0.13 μM)
WEHI-345 targets receptor-interacting protein kinase 2 (RIPK2), a serine/threonine kinase that is essential for signal transduction downstream of the nucleotide-binding oligomerization domain (NOD) receptors NOD1 and NOD2. Upon NOD stimulation, RIPK2 undergoes ubiquitination and activates NF-κB and MAPK pathways, leading to the production of pro-inflammatory cytokines. WEHI-345 inhibits RIPK2 kinase activity, thereby delaying RIPK2 ubiquitylation and NF-κB activation. It is selective for RIPK2 over RIPK1, RIPK4, and RIPK5, with Kds of 46, >10,000, >10,000, and >10,000 nM, respectively. |
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| ln Vitro |
In cells, MDP-induced RIPK2 autophosphorylation activity can be inhibited by WEHI-345 (500 nM; Raw 267.4 cells) [1]. In bone marrow, MDP-induced transcription of the inflammatory mediators TNF and interleukin 6 (IL-6) is efficiently blocked by WEHI-345 (500 nM; 0 hours, 2 hours, 4 hours, and 8 hours; BMDM or THP-1 cells). ..derived macrophages (BMDM). WEHI-345 lowers the mRNA levels of NF-kB targets in THP-1 cells, including TNF, IL-8, IL-1b, and A20 [1].
WEHI-345 is a potent inhibitor of RIPK2 with an IC50 of 0.13 μM in a kinase assay. It is selective for RIPK2 over a panel of 95 kinases at a concentration of 1 μM. The compound delays RIPK2 ubiquitylation and NF-κB activation upon NOD stimulation. By inhibiting RIPK2, WEHI-345 prevents the production of inflammatory cytokines such as TNF. It has been shown to reduce plasma levels of TNF in a mouse model of experimental autoimmune encephalomyelitis (EAE). |
| ln Vivo |
WEHI-345 (20 mg/kg; i.p.; twice daily; for 6 days; male C57BL/6 mice) treatment decreased histology scores, disease scores, inflammatory infiltrates, and the recruitment of dendritic cells to inflammatory areas. It increased body weight as well as decreased levels of cytokines and chemokines, suggesting a general improvement in wild-type C57Bl/6 mice provoked with experimental autoimmune encephalomyelitis (EAE) [1].
WEHI-345 (3-10 mg/kg) reduces plasma levels of TNF and delays disease onset in a mouse model of experimental autoimmune encephalomyelitis (EAE). This demonstrates its in vivo efficacy in a model of multiple sclerosis. The compound modulates NOD signaling events while preventing inflammatory cytokine production. Its ability to inhibit RIPK2 and reduce inflammation in vivo makes it a valuable tool for studying RIPK2-driven inflammatory diseases. |
| Enzyme Assay |
The in vitro kinase assay for WEHI-345 typically involves measuring the kinase activity of recombinant human RIPK2 using a radioactive or fluorescence-based assay. The compound is incubated with the enzyme, a peptide substrate, and ATP, and the phosphorylation level is quantified to determine the IC50. Selectivity is assessed by testing the compound against a panel of other kinases at a single concentration (e.g., 1 μM).
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: Original 267.4 Cell Tested Concentrations: 500 nM Incubation Duration: Experimental Results: Inhibition of MDP-induced RIPK2 autophosphorylation activity in cells. RT-PCR[1] Cell Types: BMDM or THP-1 Cell Tested Concentrations: 500 nM Incubation Duration: 0 hrs (hours), 2 hrs (hours), 4 hrs (hours), 8 hrs (hours) Experimental Results: Blocks the transcription of MDP-induced inflammatory mediators TNF and interleukin 6 (IL-6) in BMDM. and diminished the mRNA levels of NF-kB targets in THP-1 cells. The in vitro cellular assay for WEHI-345 typically involves culturing cells that express NOD1 or NOD2, such as macrophages or epithelial cell lines. Cells are stimulated with a NOD agonist (e.g., C12-iE-DAP for NOD1 or MDP for NOD2) in the presence or absence of varying concentrations of the compound. RIPK2 ubiquitylation is assessed by immunoprecipitation and Western blotting, and NF-κB activation is measured by reporter gene assays or by assessing the phosphorylation of IκBα. Cytokine production (e.g., TNF, IL-6) is measured by ELISA. |
| Animal Protocol |
Animal/Disease Models: C57BL/6 male mice (8 weeks old) [1]
Doses: 20 mg/kg Route of Administration: intraperitoneal (ip) injection; twice (two times) daily; for 6 days Experimental Results: disease score, inflammatory infiltrate, histological score, and inflammation diminished dendritic cell recruitment to the site. and improved body weight and diminished cytokine and chemokine levels. In vivo animal studies for WEHI-345 typically involve the use of mouse models of inflammatory diseases. In the EAE model, mice are immunized with myelin oligodendrocyte glycoprotein (MOG) peptide and treated with WEHI-345 (3-10 mg/kg) via intraperitoneal injection. Disease onset and severity are monitored by clinical scoring, and plasma levels of TNF are measured. Histological analysis of the spinal cord is performed to assess immune cell infiltration and demyelination. |
| ADME/Pharmacokinetics |
WEHI-345 has a molecular weight of 401.46 g/mol and a molecular formula of C22H23N7O. It is a crystalline solid with a purity of ≥98%. The compound is soluble in DMF (2 mg/mL) and DMSO (2 mg/mL), and slightly soluble in ethanol. It should be stored at -20°C. The compound has a λmax of 252 nm. Further detailed PK parameters are not available in the public domain.
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| Toxicity/Toxicokinetics |
Specific toxicology data for WEHI-345 are not available in the public domain. The compound is intended for research use only and should be handled with appropriate laboratory safety precautions. Standard safety assessments would be required before any clinical application. Researchers should consult the safety data sheet for detailed handling and disposal information.
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| References | |
| Additional Infomation |
WEHI-345 is a potent and selective inhibitor of receptor-interacting protein kinase 2 (RIPK2). It is used as a research tool to study NOD signaling and inflammatory diseases. The compound delays RIPK2 ubiquitylation and NF-κB activation upon NOD stimulation. It has shown efficacy in a mouse model of experimental autoimmune encephalomyelitis (EAE). It has not yet entered clinical trials and is strictly for preclinical research purposes.
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| Molecular Formula |
C22H23N7O
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|---|---|
| Molecular Weight |
401.464323282242
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| Exact Mass |
401.196
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| Elemental Analysis |
C, 65.82; H, 5.77; N, 24.42; O, 3.99
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| CAS # |
1354825-58-3
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| Related CAS # |
1354825-58-3
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| PubChem CID |
56602554
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| Appearance |
White to off-white solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
709.0±60.0 °C at 760 mmHg
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| Flash Point |
382.6±32.9 °C
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| Vapour Pressure |
0.0±2.3 mmHg at 25°C
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| Index of Refraction |
1.687
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| LogP |
2.12
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
30
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| Complexity |
586
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(C1=CC=NC=C1)NCC(C)(N2N=C(C3=CC=C(C)C=C3)C4=C(N)N=CN=C42)C
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| InChi Key |
LEBSTCDZKUSVOY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C22H23N7O/c1-14-4-6-15(7-5-14)18-17-19(23)26-13-27-20(17)29(28-18)22(2,3)12-25-21(30)16-8-10-24-11-9-16/h4-11,13H,12H2,1-3H3,(H,25,30)(H2,23,26,27)
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| Chemical Name |
N-[2-[4-amino-3-(4-methylphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-2-methylpropyl]pyridine-4-carboxamide
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| Synonyms |
WEHI 345; WEHI-345; WEHI345
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO: ~25 mg/mL (~62.3 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.23 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.23 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4909 mL | 12.4545 mL | 24.9091 mL | |
| 5 mM | 0.4982 mL | 2.4909 mL | 4.9818 mL | |
| 10 mM | 0.2491 mL | 1.2455 mL | 2.4909 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.
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