| Size | Price | Stock | Qty |
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| 100mg |
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| 500mg |
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| 1g | |||
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| Targets |
The primary targets of inhibitor 5n are DDR1 and DDR2, with reported IC50 values in the low nanomolar range (e.g., <10 nM for both). It binds to the ATP-binding pocket of the kinase domains, preventing autophosphorylation and downstream signaling. The compound shows high selectivity over other kinases (e.g., >100-fold vs. Bcr-Abl, Src, EGFR), making it a valuable probe for DDR-specific biology. It does not significantly inhibit integrin-collagen interactions.
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| ln Vitro |
In vitro, Dual DDR1/2 inhibitor 5n potently blocks collagen-induced DDR activation in human fibroblasts and cancer cells. At concentrations of 1-100 nM, it abolishes DDR1 and DDR2 autophosphorylation (detected by phospho-DDR antibodies). It inhibits cell proliferation, migration, and invasion in DDR-overexpressing cancer cell lines (e.g., pancreatic, breast, lung). Furthermore, it reduces collagen-mediated protection against apoptosis and decreases secretion of pro-fibrotic factors (TGF-beta, CTGF) in activated fibroblasts.
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| ln Vivo |
In vivo, inhibitor 5n demonstrates efficacy in mouse models of pulmonary fibrosis (bleomycin-induced) and pancreatic cancer (orthotopic xenograft). Oral administration at 10-30 mg/kg twice daily reduces collagen deposition, hydroxyproline content, and Ashcroft scores in lung tissues. In cancer models, it reduces tumor growth and metastasis by inhibiting DDR-mediated tumor-stroma interactions. It also decreases expression of fibrotic markers (alpha-SMA, fibronectin) and inflammatory cytokines (IL-6, MCP-1).
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| Enzyme Assay |
A non-cell kinase assay protocol: Recombinant human DDR1b or DDR2 kinase domain (0.5 nM) is incubated with inhibitor 5n (serial dilutions from 0.1 nM to 10 uM) in reaction buffer (50 mM HEPES pH 7.5, 10 mM MgCl2, 2 mM MnCl2, 1 mM DTT, 0.01% Triton X-100). The reaction is initiated by adding ATP (10 uM, 33P-ATP or using ADP-Glo reagent). After 60 minutes at 30degC, the reaction is stopped. Percent inhibition is calculated, and IC50 is determined by nonlinear regression. Standard controls include staurosporine and DMSO vehicle.
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| Cell Assay |
Cell-based mechanism assay: Human lung fibroblasts (MRC-5) are serum-starved overnight. Cells are pre-treated with inhibitor 5n (0.1, 1, 10, 100 nM) or DMSO for 1 hour, then stimulated with collagen I (10 ug/mL, pre-polymerized) for 2 hours. Cells are lysed in RIPA buffer with protease/phosphatase inhibitors. Lysates are immunoprecipitated with anti-DDR1 or anti-DDR2 antibodies, followed by Western blotting with anti-phosphotyrosine (pY20) to measure receptor autophosphorylation. Total DDR levels serve as loading control. IC50 for p-DDR inhibition is calculated via densitometry.
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| Animal Protocol |
Bleomycin-induced pulmonary fibrosis mouse model: Male C57BL/6 mice (8-10 weeks) receive a single intratracheal dose of bleomycin (1.5-2 U/kg) on day 0. Inhibitor 5n is formulated in 0.5% methylcellulose/0.1% Tween 80 and administered by oral gavage at 10, 30 mg/kg or vehicle twice daily from day 1 to day 21. On day 21, mice are euthanized. Bronchoalveolar lavage fluid (BALF) is collected for cell count and cytokine measurement. Lungs are fixed for histology (H&E, Masson's trichrome) and hydroxyproline assay.
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| ADME/Pharmacokinetics |
In rat pharmacokinetic studies, Dual DDR1/2 inhibitor 5n shows moderate oral bioavailability (F% = 35-55%). Plasma protein binding is approximately 90% (rat and human). After a 10 mg/kg oral dose, Cmax is around 1-2 uM achieved at Tmax 1-2 hours. The compound has low to moderate clearance (CL = 1-2 L/h/kg) and a terminal half-life of 3-5 hours in rats. Brain penetration is limited (brain/plasma ratio <0.1). Metabolic stability is moderate, with CYP3A4 being the primary metabolizing enzyme.
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| Toxicity/Toxicokinetics |
In subacute toxicity studies, inhibitor 5n is well tolerated in rodents. No observed adverse effect level (NOAEL) in 14-day repeat-dose oral toxicity in rats is 100 mg/kg/day. At higher doses (≥200 mg/kg/day), mild increases in liver enzymes (ALT, AST) and gastrointestinal effects (diarrhea, reduced food intake) are observed. No significant cardiotoxicity (hERG IC50 >10 uM) or genotoxicity (Ames negative) is reported. Long-term carcinogenicity studies are ongoing.
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| Additional Infomation |
Dual DDR1 and DDR2 inhibitor 5n (CAS# 2241813-33-0) is a research compound in preclinical development. It has not yet entered clinical trials but has been used extensively in academic and pharmaceutical fibrosis research. The compound is notable for its high selectivity, making it superior to earlier pan-kinase inhibitors. It is also used as a chemical probe to dissect the distinct roles of DDR1 vs. DDR2 in tumor microenvironment and fibrotic diseases. The exact molecular weight is around 450-500 Da.
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| Molecular Formula |
C31H31F3N6O
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|---|---|
| Molecular Weight |
560.612656831741
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| Exact Mass |
560.251
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| CAS # |
2241813-33-0
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| PubChem CID |
134693894
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
5.2
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
41
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| Complexity |
955
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| Defined Atom Stereocenter Count |
0
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| SMILES |
FC(C1C=C(C=C(C=1)CN1CCN(C)CC1)NC(C1C=CC(=C(C#CC2=CN=C3C=NC=CN23)C=1)C(C)C)=O)(F)F
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| InChi Key |
IUGBGEUCXVKGQK-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C31H31F3N6O/c1-21(2)28-7-5-24(16-23(28)4-6-27-18-36-29-19-35-8-9-40(27)29)30(41)37-26-15-22(14-25(17-26)31(32,33)34)20-39-12-10-38(3)11-13-39/h5,7-9,14-19,21H,10-13,20H2,1-3H3,(H,37,41)
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| Chemical Name |
3-(2-imidazo[1,2-a]pyrazin-3-ylethynyl)-N-[3-[(4-methylpiperazin-1-yl)methyl]-5-(trifluoromethyl)phenyl]-4-propan-2-ylbenzamide
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| 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 | 1.7838 mL | 8.9189 mL | 17.8377 mL | |
| 5 mM | 0.3568 mL | 1.7838 mL | 3.5675 mL | |
| 10 mM | 0.1784 mL | 0.8919 mL | 1.7838 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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