| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg | |||
| 100mg | |||
| 500mg | |||
| 1g | |||
| Other Sizes |
| Targets |
BRAF (B-Raf proto-oncogene, serine/threonine kinase), specifically BRAFWT and the constitutively active BRAFV600E mutant. B-Raf IN 15 is an ATP-competitive kinase inhibitor. It binds to the ATP-binding pocket of BRAF, blocking its kinase activity. This disrupts the downstream signaling of the MAPK/ERK pathway (Raf-MEK-ERK), which is critical for cell proliferation and survival. It is a potent inhibitor with IC50 values of 2.0 microM for BRAFWT and 0.8 microM for BRAFV600E.
|
|---|---|
| ln Vitro |
B-Raf IN 15 inhibits BRAFWT and BRAFV600E. The in vitro potency is determined by biochemical assays, showing IC50s of 2.0 microM for the wild-type (WT) enzyme and 0.8 microM for the V600E mutant. This indicates it is more potent against the oncogenic mutant form. It can be used to study cancer and can be optimized for use as a more effective BRAF inhibitor in melanoma and other cancers.
|
| ln Vivo |
No specific in vivo data has been reported. As a BRAF inhibitor, it is expected to demonstrate antitumor efficacy in mouse xenograft models of BRAFV600E-mutant melanoma (e.g., A375) and colorectal cancer. The antitumor effect would be accompanied by the suppression of phosphorylated ERK (pERK) levels in the tumors. It would likely be administered orally.
|
| Enzyme Assay |
B-Raf IN 15 is a potent BRAFWT and BRAFV600E inhibitor. A typical cell-free assay for kinase inhibition is performed using purified recombinant human BRAFWT or BRAFV600E protein. The enzyme (5-10 nM) is incubated with various concentrations of B-Raf IN 15 (0.1 nM-100 uM) in a reaction buffer (25 mM HEPES, pH 7.5, 10 mM MgCl2, 1 mM DTT) for 30 minutes at room temperature. The reaction is started by adding a substrate (inactive MEK1 protein) and 10 microM ATP. After 60 minutes at 30degC, the reaction is stopped with SDS-PAGE loading buffer. The amount of phosphorylated MEK1 is quantified by Western blotting using a specific anti-phospho-MEK antibody, or by using an ELISA-based detection kit. The IC50 value is calculated by plotting the percentage of inhibition against the log of the inhibitor concentration.
|
| Cell Assay |
For in vitro cellular studies, BRAFV600E-mutant human melanoma cell lines (e.g., A375, SK-MEL-28) are seeded in 96-well plates (3×103 cells/well). After 24 hours, cells are treated with B-Raf IN 15 at concentrations ranging from 0.01-100 uM for 48-72 hours. Cell viability is assessed using the CellTiter-Glo (ATP) assay or the MTT assay. The GI50 (50% growth inhibitory concentration) is determined. For mechanistic studies, cells are treated with the compound for 2-6 hours, and cell lysates are prepared. The inhibition of the MAPK pathway is assessed by Western blotting for phosphorylated MEK (pMEK) and phosphorylated ERK (pERK). Apoptosis is measured by caspase-3/7 activity.
|
| Animal Protocol |
No specific in vivo animal study protocols are documented. Based on standard BRAF inhibitor study designs, a typical protocol would involve establishing subcutaneous xenografts of BRAFV600E-mutant A375 melanoma cells in 6-8 week old female athymic nude mice. When tumors reach approximately 150-200 mm3, mice are randomized into treatment groups (n=8-10 per group). B-Raf IN 15 would be administered by oral gavage at doses of 10-50 mg/kg, once daily or twice daily, for 14-21 days. Tumor volume is measured with calipers every 2-3 days. At the end of the study, tumors are excised, weighed, and analyzed for pERK levels by IHC and Western blot to confirm target engagement. Body weight is monitored as a measure of tolerability.
|
| ADME/Pharmacokinetics |
No pharmacokinetic data has been reported. As a small molecule with a molecular weight of 333.41 g/mol, and given that it is a kinase inhibitor, it is likely to be orally bioavailable. However, its specific ADME (absorption, distribution, metabolism, excretion) profile has not been published. It likely has a short to moderate half-life (2-8 hours) and is metabolized by liver CYP450 enzymes, particularly CYP3A4.
|
| Toxicity/Toxicokinetics |
No specific toxicity data has been reported. However, BRAF inhibitors as a class have a well-defined toxicity profile. These include dermatologic toxicities (rash, photosensitivity, and the development of cutaneous squamous cell carcinomas due to paradoxical activation of the MAPK pathway in normal cells), arthralgia (joint pain), fatigue, and gastrointestinal issues. The severity of these effects is dose-dependent and manageable but requires monitoring.
|
| References | |
| Additional Infomation |
B-Raf IN 15 (Compound 7) is a small molecule inhibitor of the BRAF kinase (wild-type and V600E mutant). It is a chemical probe for studying the MAPK signaling pathway in cancer biology. The V600E mutation is the most common activating mutation in melanoma. The molecular formula is C19H15N3OS, and the molecular weight is 333.41 g/mol. It is for research use only, not approved for clinical therapy. Its IUPAC name is 2-chloro-5-[3-(3,5-dimethylpyrazol-1-yl)-2-oxo-1,3-diazaspiro[4.5]dec-1-yl]benzoic acid.
|
| Molecular Formula |
C19H15N3OS
|
|---|---|
| Molecular Weight |
333.407
|
| Exact Mass |
333.09
|
| Elemental Analysis |
C, 68.45; H, 4.53; N, 12.60; O, 4.80; S, 9.62
|
| CAS # |
832107-31-0
|
| PubChem CID |
2977311
|
| Appearance |
Off-white to light yellow solid powder
|
| LogP |
3.6
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
24
|
| Complexity |
410
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C(C1=CSC=C1)(C1C=CC2C=CC=NC=2C=1O)NC1=CN=CC=C1
|
| InChi Key |
KSWFNFIKRRTNCX-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C19H15N3OS/c23-19-16(6-5-13-3-1-9-21-18(13)19)17(14-7-10-24-12-14)22-15-4-2-8-20-11-15/h1-12,17,22-23H
|
| Chemical Name |
7-((pyridin-3-ylamino)(thiophen-3-yl)methyl)quinolin-8-ol
|
| Synonyms |
B-Raf IN 15; B-Raf IN-15; B Raf IN 15; Compound 7
|
| 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 (~299.93 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.50 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (7.50 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9993 mL | 14.9966 mL | 29.9931 mL | |
| 5 mM | 0.5999 mL | 2.9993 mL | 5.9986 mL | |
| 10 mM | 0.2999 mL | 1.4997 mL | 2.9993 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
