| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
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| 5mg | |||
| 100mg | |||
| Other Sizes |
| Targets |
c-Src (Ki = 44 nM)
c-Raf (>95% displacement at 10 µM in binding assay) B-Raf (>95% displacement at 10 µM in binding assay) Lck (Kd = 160 nM) Fgr (Kd = 240 nM) Yes (Kd = 720 nM) Lyn (Kd = 3200 nM) Hck (Kd = 4400 nM) Fyn (Kd > 40,000 nM)[1] |
|---|---|
| ln Vitro |
At concentrations up to 125 μM, KB SRC 4 (Compound 4) exhibits no inhibitory action on c-Abl and is a strong, highly selective c-Src inhibitor with a Ki of 44 nM and a Kd of 86 nM. Eight times more selective than c-Yes (Kd, 720 nM), KB SRC 4 is selective among Src family members, two times more selective than Lck (Kd, 160 nM) and Fgr (Kd, 240 nM), and forty times more selective than Lyn (Kd, 3200 nM), Hck (Kd, 4400 nM), and Fyn (Kd, >40000 nM). With growth inhibition coefficients (GI50s) of 11 μM, 12 μM, 11 μM, and 6.0 μM for HT-29, SK-BR-3, MCF7, MDA-MB-453, and NIH-3T3, respectively, KB SRC 4 is effective against cancer cell lines [1].
KB SRC 4 inhibits c-Src activity in murine embryonal fibroblast (MEF) cells expressing full-length c-Src, with an IC50 of 1.9 µM in a cellular phosphorylation assay measuring phosphorylation of c-Src Tyr-416.[1] KB SRC 4 shows potent growth inhibition in multiple cancer cell lines dependent on c-Src activity, including HT-29 (GI50 = 11 µM), SK-BR-3 (GI50 = 12 µM), MCF7 (GI50 = 11 µM), and MDA-MB-453 (GI50 = 6.0 µM).[1] KB SRC 4 does not inhibit non-cancerous NIH-3T3 cell growth up to 100 µM, whereas PP2 shows GI50 of 17 µM in the same cell line.[1] In 4T1 breast cancer cells cultured in 3D, KB SRC 4 (10 µM) significantly inhibits cell growth, an effect that is mitigated by co-treatment with the c-Abl inhibitor GNF-2.[1] KB SRC 4 does not affect phospho-Erk levels in SK-BR-3 cells stimulated with EGF, suggesting no significant cellular inhibition of Raf kinases.[1] In the NCI 60 panel, KB SRC 4 (10 µM) shows cytostatic activity with mean growth of 71% across 57 cell lines, and seven cell lines showed <50% growth.[1] |
| Enzyme Assay |
A continuous fluorimetric assay was used to determine inhibitor efficacy. Phosphorylation of a self-reporting peptide substrate leads to an increase in fluorescence emission at 405 nm (excitation at 340 nm). Final assay concentrations were ATP = 100 µM and substrate peptide = 45 µM. Dose-response curves were generated for each compound to determine Ki values.[1]
The binding affinity of KB SRC 4 to c-Src and other Src family kinases was measured using an in vitro ATP-site competition binding assay (KINOMEscan) at a concentration of 10 µM, and Kd values were determined for several kinases.[1] |
| Cell Assay |
For cellular phosphorylation assays, murine embryonal fibroblast (MEF) cells exogenously expressing full-length c-Src were incubated with KB SRC 4. Phosphorylation of c-Src Tyr-416 was measured using specific antibodies in a sandwich ELISA format.[1]
For cell growth inhibition assays, cancer cell lines were plated in 96-well microtiter plates and allowed to attach for 24 hours. Compounds were added in medium containing 1% DMSO and incubated for 72 hours. Cell viability was assessed using a tetrazolium-based reagent, and absorbance was measured at 450 nm and 630 nm. Data were analyzed using curve-fitting software.[1] For phospho-Erk assays, SK-BR-3 cells were plated in 96-well plates, serum-starved overnight, treated with compound for 60 minutes, stimulated with EGF, and lysed. Lysates were analyzed using a commercial assay kit for phosphorylated Erk1/2.[1] For 4T1 cell growth in 3D culture, cells were cultured on a basement membrane extract cushion in growth media supplemented with 5% basement membrane extract. After 72 hours, compounds were added and incubated for an additional 72 hours. Cell viability was measured using a tetrazolium-based reagent.[1] |
| Toxicity/Toxicokinetics |
KB SRC 4 did not show growth inhibition at concentrations up to 100 µM in non-cancerous NIH-3T3 cells, indicating reduced toxicity compared to PP2. [1]
|
| References | |
| Additional Infomation |
KB SRC 4 is a highly selective and cell-penetrating c-Src inhibitor that works by extending into the phosphate-binding loop (P-loop) pocket, a strategy designed to enhance the selectivity of kinase inhibitors. [1] Compared to multi-kinase inhibitors, KB SRC 4 selectively inhibits c-Src more effectively to slow cancer cell growth and avoids inhibiting anti-cancer kinases such as c-Abl, which is advantageous in breast cancer models. [1] In the NCI 60 cell line, KB SRC 4 exhibits cytotoxic rather than cytotoxic effects, consistent with the genomic repressive effects of c-Src. [1]
|
| Molecular Formula |
C32H23N8CL
|
|---|---|
| Molecular Weight |
555.03162
|
| Exact Mass |
554.173
|
| CAS # |
1380088-03-8
|
| PubChem CID |
73349077
|
| Appearance |
Light brown to brown solid powder
|
| LogP |
7.273
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
41
|
| Complexity |
835
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
UHIZYQVRKSWIFO-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C32H23ClN8/c33-26-14-12-23(13-15-26)30-29-31(34)35-20-36-32(29)41(38-30)27-11-5-10-25(17-27)28-18-37-39-40(28)19-21-6-4-9-24(16-21)22-7-2-1-3-8-22/h1-18,20H,19H2,(H2,34,35,36)
|
| Chemical Name |
3-(4-chlorophenyl)-1-[3-[3-[(3-phenylphenyl)methyl]triazol-4-yl]phenyl]pyrazolo[3,4-d]pyrimidin-4-amine
|
| 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 (~180.17 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 | 1.8017 mL | 9.0085 mL | 18.0170 mL | |
| 5 mM | 0.3603 mL | 1.8017 mL | 3.6034 mL | |
| 10 mM | 0.1802 mL | 0.9009 mL | 1.8017 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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