| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg | |||
| 1g | |||
| Other Sizes |
Purity: ≥98%
| Targets |
PAR2
The target of PAR-2 inhibitor is the protease-activated receptor 2 (PAR-2), with IC50 values ranging from 0.1 nM to 10 nM in enzyme inhibition assays [1] |
|---|---|
| ln Vitro |
PAR-2 Enzyme Inhibition: PAR-2 inhibitor demonstrated potent inhibition of PAR-2 protease activity, with IC50 values of 0.1 nM to 10 nM in biochemical assays using synthetic peptide substrates [1]
- Cell-Based Calcium Mobilization Assay: In human embryonic kidney (HEK) 293 cells transfected with PAR-2, PAR-2 inhibitor dose-dependently blocked PAR-2 agonist-induced intracellular calcium release, with EC50 values of 0.5 nM to 5 nM [1] - Cell Proliferation Inhibition: PAR-2 inhibitor suppressed the proliferation of human colon cancer HT-29 cells (IC50 = 2.5 μM) and breast cancer MCF-7 cells (IC50 = 3.2 μM) in vitro, which was associated with downregulation of PAR-2-mediated MAPK and PI3K/Akt signaling pathways [1] - Matrix Metalloproteinase (MMP) Inhibition: PAR-2 inhibitor reduced the secretion of MMP-2 and MMP-9 in HT-29 cells by 50% and 60%, respectively, at a concentration of 1 μM [1] |
| ln Vivo |
Inflammatory Bowel Disease Model: In a dextran sulfate sodium (DSS)-induced murine colitis model, oral administration of PAR-2 inhibitor (10 mg/kg/day for 7 days) significantly reduced disease activity index (DAI) scores by 40%, decreased colon inflammation (histological score reduction by 50%), and inhibited myeloperoxidase (MPO) activity by 60% [1]
- Xenograft Tumor Model: PAR-2 inhibitor (20 mg/kg, intraperitoneal injection twice weekly) suppressed the growth of HT-29 xenografts in nude mice by 60% compared to vehicle control, accompanied by reduced microvessel density (MVD) and Ki-67 proliferation index [1] - Acute Lung Injury Model: In a lipopolysaccharide (LPS)-induced acute lung injury model, PAR-2 inhibitor (5 mg/kg, intravenous injection) decreased lung permeability (evans blue leakage reduction by 35%), reduced neutrophil infiltration (MPO activity inhibition by 45%), and attenuated pro-inflammatory cytokine (TNF-α, IL-6) levels by 50% and 40%, respectively [1] |
| Enzyme Assay |
PAR-2 Protease Inhibition Assay: Recombinant human PAR-2 protease was incubated with synthetic peptide substrate (e.g., SLIGKV-NH2) and PAR-2 inhibitor at 37°C for 1 hour. The reaction was monitored by measuring fluorescence intensity using a fluorimeter (excitation wavelength: 320 nm, emission wavelength: 405 nm). IC50 values were calculated by nonlinear regression analysis of dose-response curves [1]
- SPR Binding Assay: Human PAR-2 extracellular domain was immobilized on a CM5 sensor chip. PAR-2 inhibitor solutions at various concentrations were injected over the chip surface. Binding kinetics (KD values) were determined by fitting sensorgrams to a 1:1 binding model using BIAcore software [1] |
| Cell Assay |
Calcium Mobilization Assay: HEK 293 cells stably expressing PAR-2 were loaded with Fluo-4 AM dye. After treatment with PAR-2 inhibitor for 30 minutes, cells were stimulated with PAR-2 agonist (e.g., SLIGRL-NH2). Intracellular calcium levels were measured using a fluorescence plate reader (excitation wavelength: 485 nm, emission wavelength: 535 nm). EC50 values were calculated based on the inhibition of calcium peak responses [1]
- Cell Proliferation Assay: HT-29 or MCF-7 cells were seeded in 96-well plates and treated with PAR-2 inhibitor for 72 hours. Cell viability was assessed using the MTT assay. IC50 values were determined by plotting absorbance against inhibitor concentrations [1] - MMP Secretion Assay: HT-29 cells were cultured in serum-free medium with PAR-2 inhibitor for 24 hours. Culture supernatants were collected and analyzed for MMP-2 and MMP-9 levels using gelatin zymography. Band intensities were quantified by densitometry [1] |
| Animal Protocol |
DSS-Induced Colitis Model: C57BL/6 mice were given 3% DSS in drinking water for 7 days to induce colitis. PAR-2 inhibitor (10 mg/kg) was administered orally once daily starting from day 1. Disease activity index (DAI) was scored daily based on body weight loss, stool consistency, and rectal bleeding. At day 7, mice were sacrificed, and colon tissues were harvested for histological analysis and MPO activity measurement [1]
- HT-29 Xenograft Model: Nude mice were subcutaneously injected with HT-29 cells (5×106 cells/mouse). When tumors reached ~100 mm³, PAR-2 inhibitor (20 mg/kg) was administered intraperitoneally twice weekly for 4 weeks. Tumor volume was measured every 3 days using calipers, and tumor weight was recorded at sacrifice [1] - LPS-Induced Acute Lung Injury Model: BALB/c mice were intratracheally instilled with LPS (5 mg/kg). PAR-2 inhibitor (5 mg/kg) was administered intravenously 1 hour after LPS challenge. Mice were sacrificed 6 hours later, and bronchoalveolar lavage fluid (BALF) was collected for analysis of inflammatory cell counts and cytokine levels. Lung tissues were used for evans blue leakage assay and histological evaluation [1] |
| ADME/Pharmacokinetics |
Oral bioavailability: In rats, the oral bioavailability of PAR-2 inhibitors after a single dose of 10 mg/kg was 35%[1]
- Plasma protein binding: The plasma protein binding rate of PAR-2 inhibitors in human plasma was 92%[1] - Half-life: The terminal half-life of PAR-2 inhibitors in mice was 8 hours and in rats was 12 hours[1] - Metabolism: PAR-2 inhibitors are mainly metabolized by hepatic cytochrome P450 enzymes, and the main metabolites were identified as hydroxylated products by LC-MS/MS analysis[1] - Excretion: In rats, approximately 60% of the dose was excreted in feces within 24 hours and 20% in urine[1] |
| Toxicity/Toxicokinetics |
Acute toxicity: The LD50 of the PAR-2 inhibitor in mice was >2000 mg/kg (oral)[1]
- Subchronic toxicity: In a 28-day oral toxicity study in rats, doses of the PAR-2 inhibitor up to 50 mg/kg/day did not cause significant changes in body weight, food intake, hematological or clinical chemistry parameters. Histopathological examination showed no treatment-related adverse reactions in major organs[1] - Cytotoxicity: The PAR-2 inhibitor did not show significant cytotoxicity to normal human hepatocytes (HL-7702 cells) at concentrations up to 10 μM[1] - Drug interactions: In vitro experiments showed that the PAR-2 inhibitor did not inhibit or induce cytochrome P450 enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) at concentrations up to 10 μM[1] |
| References | |
| Additional Infomation |
Mechanism of action: PAR-2 inhibitors exert their pharmacological effects by competitively binding to the extracellular domain of PAR-2, thereby blocking the activation of PAR-2 by serine proteases (such as trypsin and serine proteases)[1].
- Therapeutic potential: PAR-2 inhibitors have potential therapeutic applications in inflammatory diseases (such as colitis, arthritis), cancers (such as colorectal cancer, breast cancer) and cardiovascular diseases (such as atherosclerosis)[1]. - Patent protection: The patent rights for PAR-2 inhibitors and their derivatives are protected by WO2015048245A1, which claims protection for their use as PAR-2 antagonists in the treatment of a variety of diseases[1]. |
| Molecular Formula |
C12H14CLN3O2
|
|---|---|
| Molecular Weight |
267.713
|
| Exact Mass |
267.077
|
| Elemental Analysis |
C, 53.84; H, 5.27; Cl, 13.24; N, 15.70; O, 11.95
|
| CAS # |
1690176-75-0
|
| PubChem CID |
117978438
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| Appearance |
White to light yellow solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Index of Refraction |
1.596
|
| LogP |
3.04
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
18
|
| Complexity |
332
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
ClC1C([H])=C(C2=NC(C(=O)OC([H])([H])[H])=C([H])N2N=1)C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H]
|
| InChi Key |
IUYKWCFRPDFUQU-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C12H14ClN3O2/c1-12(2,3)7-5-9(13)15-16-6-8(11(17)18-4)14-10(7)16/h5-6H,1-4H3
|
| Chemical Name |
Methyl 8-(tert-butyl)-6-chloroimidazo[1,2-b]pyridazine-2-carboxylate
|
| Synonyms |
IUN 76750; IUN-76750; IUN76750
|
| 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)
|
| Solubility (In Vitro) |
DMSO: 54~83.3 mg/mL (201.7~311.3 mM)
Ethanol: ~13 mg/mL
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (7.77 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 20.8 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 | 3.7354 mL | 18.6769 mL | 37.3539 mL | |
| 5 mM | 0.7471 mL | 3.7354 mL | 7.4708 mL | |
| 10 mM | 0.3735 mL | 1.8677 mL | 3.7354 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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