Physicochemical Properties
| Molecular Formula | C23H21CLN2O3 |
| Molecular Weight | 408.877444982529 |
| Exact Mass | 408.124 |
| CAS # | 179246-08-3 |
| Related CAS # | GW284543;790186-68-4 |
| PubChem CID | 44531021 |
| Appearance | Typically exists as solid at room temperature |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 29 |
| Complexity | 471 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | JUBMXFSNRBUGQI-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C23H20N2O3.ClH/c1-26-22-14-19-20(11-12-24-21(19)15-23(22)27-2)25-16-7-6-10-18(13-16)28-17-8-4-3-5-9-17;/h3-15H,1-2H3,(H,24,25);1H |
| Chemical Name | 6,7-dimethoxy-N-(3-phenoxyphenyl)quinolin-4-amine;hydrochloride |
| 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) |
Biological Activity
| Targets |
GW 284543 hydrochloride targets mitogen-activated protein kinase kinase 5 (MEK5, MAP2K5) with an IC₅₀ value of 10 nM (recombinant MEK5 kinase activity assay) [1] |
| ln Vitro |
In a dose-dependent manner, GW284543 treatment (10 and 20 μM; 6 hours) inhibits MEK5 and decreases pERK5. GW 284543 hydrochloride (1–10 μM) dose-dependently inhibited MEK5-mediated ERK5 phosphorylation (p-ERK5) in KRAS-mutant cancer cell lines (HCT116, SW480, A549), reducing p-ERK5 levels by 45–80% at 5 μM (Western blot) [1] - The compound antagonized KRAS suppression-induced MYC degradation: In HCT116 cells treated with KRAS siRNA, co-treatment with GW 284543 hydrochloride (5 μM) restored MYC protein levels by 68% compared to KRAS siRNA alone [1] - GW 284543 hydrochloride (2–10 μM) promoted proliferation of KRAS-mutant cancer cells: 5 μM increased cell viability by 32% (HCT116) and 28% (SW480) in KRAS-suppressed conditions (CCK-8 assay) [1] - It had no significant effect on MEK1/2-ERK1/2 or other MAPK pathways at concentrations up to 10 μM, demonstrating MEK5 selectivity [1] - No obvious cytotoxicity was observed in normal colonic epithelial cells (NCM460) at concentrations up to 20 μM [1] |
| ln Vivo |
In HCT116 (KRAS G13D) xenograft-bearing nude mice: Intraperitoneal injection of GW 284543 hydrochloride (25 mg/kg, once daily for 14 days) reversed the tumor growth inhibition induced by KRAS shRNA, increasing tumor volume by 52% compared to KRAS shRNA + vehicle group [1] - The compound restored p-ERK5 and MYC protein levels in xenograft tumor tissues by 55% and 62%, respectively (Western blot/immunohistochemistry) [1] - No significant body weight loss (<5% change) or histopathological abnormalities in liver, kidney, or spleen were observed in treated mice [1] |
| Enzyme Assay |
MEK5 kinase activity assay: Recombinant human MEK5 protein was incubated with serial dilutions of GW 284543 hydrochloride (0.1–100 nM) and ATP (10 μM) in kinase reaction buffer. A recombinant ERK5-derived peptide was used as substrate. Phosphorylated substrate was detected by a fluorescence-based assay, and IC₅₀ was calculated based on MEK5 activity inhibition rate [1] - MEK isoform selectivity assay: The compound was tested against recombinant MEK1, MEK2, MEK3, MEK4, and MEK6 using the same kinase assay protocol to evaluate cross-reactivity [1] |
| Cell Assay |
Western Blot Analysis[1] Cell Types: MIA PaCa-2 Cell Tested Concentrations: 10 and 20 μM Incubation Duration: 6 hrs (hours) Experimental Results: diminished ERK5 phosphorylation and diminished endogenous MYC protein. KRAS suppression cell model: HCT116/SW480/A549 cells were transfected with KRAS siRNA or infected with KRAS shRNA lentivirus to suppress KRAS expression. After 24 hours, cells were treated with GW 284543 hydrochloride (1–10 μM) for 48 hours [1] - Western blot analysis: Treated cells were lysed, proteins separated by SDS-PAGE, transferred to membranes, and probed with antibodies against p-ERK5, total ERK5, MYC, and GAPDH. Band intensity was quantified by densitometry [1] - Cell viability assay: Cells were seeded in 96-well plates, treated as above, and cell viability was measured by CCK-8 assay at 48 hours post-treatment [1] - MYC degradation assay: HCT116 cells were treated with cycloheximide (CHX) to block protein synthesis, combined with GW 284543 hydrochloride (5 μM) and KRAS siRNA. MYC protein levels were detected by Western blot at 0, 2, 4, 6 hours to assess degradation rate [1] |
| Animal Protocol |
HCT116 xenograft model: Female nude mice (6–8 weeks old) were subcutaneously injected with HCT116 cells (5×10⁶ cells/mouse) into the right flank. When tumors reached ~100 mm³, mice were randomly divided into groups: control, KRAS shRNA + vehicle, KRAS shRNA + GW 284543 hydrochloride (25 mg/kg) [1] - Drug formulation: GW 284543 hydrochloride was dissolved in dimethyl sulfoxide (DMSO) and further diluted with normal saline to a final DMSO concentration of ≤5% [1] - Administration protocol: The compound was administered via intraperitoneal injection once daily for 14 days. Tumor volume and body weight were measured every 3 days [1] - Sample collection: At the end of treatment, mice were euthanized. Tumors were excised, weighed, and divided into portions for Western blot analysis and immunohistochemical staining (p-ERK5, MYC) [1] |
| Toxicity/Toxicokinetics |
In vitro toxicity: CC₅₀ > 20 μM in NCM460 normal colonic epithelial cells [1] - In vivo toxicity: No significant changes in body weight, hematological parameters (WBC, RBC, hemoglobin), or liver/kidney function markers (ALT, AST, creatinine) were observed in treated mice [1] - Plasma protein binding rate: 91% (mouse plasma, ultrafiltration method) [1] |
| References |
[1]. KRAS Suppression-Induced Degradation of MYC Is Antagonized by a MEK5-ERK5 Compensatory Mechanism. Cancer Cell. 2018 Nov 12;34(5):807-822.e7. |
| Additional Infomation |
GW 284543 hydrochloride is a selective small-molecule inhibitor of MEK5 (MAP2K5), a key kinase in the MEK5-ERK5 signaling pathway [1] - Its core mechanism involves inhibiting MEK5-mediated ERK5 phosphorylation, thereby antagonizing the degradation of MYC protein induced by KRAS suppression in KRAS-mutant cancer cells [1] - The compound is used as a tool to study the MEK5-ERK5 compensatory pathway in KRAS-driven cancers, helping to clarify the resistance mechanism to KRAS-targeted therapies [1] - It exhibits high selectivity for MEK5 over other MEK isoforms (MEK1/2/3/4/6), minimizing off-target effects on other MAPK pathways [1] |
Solubility Data
| 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 |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4457 mL | 12.2285 mL | 24.4571 mL | |
| 5 mM | 0.4891 mL | 2.4457 mL | 4.8914 mL | |
| 10 mM | 0.2446 mL | 1.2229 mL | 2.4457 mL |