| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg | |||
| 10mg | |||
| Other Sizes |
| Targets |
M443 targets MRK (MAPKAP kinase, also known as MAPKAPK5). It acts as an irreversible, specific inhibitor with an IC50 of <125 nM. By inhibiting MRK, the compound blocks the radiation-induced activation of downstream targets p38 MAPK and Chk2.
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| ln Vitro |
MRK depletion decreased cell survival to 33% of control after 3 Gy of IR. Similarly, at 10% viability, clonogenic experiments demonstrated a significant reduction of survival at a dosage enhancement factor (DEF) of 1.6. The peak of IR-induced MRK activation occurs 30 minutes following radiation exposure. Consequently, this time point was selected for further analysis. 500 nM M443 significantly reduced the activation of MRK, Chk2, and p38 in response to IR in both cell cultures. The following procedures were used to prepare the cells for immunofluorescence: they were seeded on coverslips, pretreated with 500 nM M443 or vehicle, subjected to 6 Gy IR, frozen at different intervals after IR, and processed using an MPM2 phospho-specific antibody that specifically detects mitotic cells. M443-treated cells did not arrest following IR, in contrast to control cells, and they retained a mitotic index that was comparable to that of non-irradiated cells. As a result, blocking MRK inhibits Chk2 and does not stop the cell cycle reaction to damage to DNA caused by IR [1].
M443 potently inhibits MRK with an IC50 of <125 nM. It inhibits radiation-induced activation of p38 and Chk2. The compound strongly radiosensitizes UW228 medulloblastoma cells and UI226 patient-derived primary cells, while it does not affect the response to radiation of normal brain cells. |
| ln Vivo |
The average survival time of control mice following tumor cell implantation was 32 days. Only M443 treatment increased survival by 5.5 days, while certain low-dose radiation did not show a discernible improvement in survival. On the other hand, IR and M443 together increased survival; the median survival increased by 16 days when compared to the control group. Animal body weight was not impacted by M443 treatment, as weight loss was seen in all groups in the days preceding the animals' demise. The findings demonstrated that tumor-containing regions (lane RB in the brain injected with a vehicle) had higher levels of both total and active MRK. MRK activity was completely absent from tumor-containing regions of the brain that were treated with M443. Curiously, normal brain areas that had some MRK protein in control brains likewise lost MRK in treated brains, indicating that normal MRK is suppressed by M443 spreading across the cerebellum [1].
In vivo, M443 has been studied for its ability to radiosensitize tumors. The compound inhibits radiation-induced activation of p38 and Chk2. Its potential in cancer research, particularly in combination with radiation therapy, has been investigated. |
| Enzyme Assay |
In vitro enzyme assays measure the inhibition of MRK kinase activity. M443 is incubated with purified MRK and a substrate, and kinase activity is measured using radiolabeled ATP or phospho-specific antibodies. The IC50 is determined from dose-response curves. Irreversible inhibition is confirmed by dialysis experiments.
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| Cell Assay |
In vitro cellular assays evaluate the effects of M443 on DNA damage response pathways. Cells are treated with M443 and exposed to radiation. The phosphorylation of p38 and Chk2 is measured by Western blotting. Cell survival and radiosensitization are assessed using clonogenic assays.
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| Animal Protocol |
In vivo animal experiments are conducted in mouse xenograft models of medulloblastoma. M443 is administered and tumors are irradiated. Tumor growth inhibition and radiation response are assessed. The compound's ability to radiosensitize tumors without affecting normal tissues is evaluated.
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| ADME/Pharmacokinetics |
M443 has a CAS number of 1820684-31-8. It is soluble in DMSO at 55 mg/mL. The compound is an irreversible, specific inhibitor of MRK. It is also known as EVT-274557. The compound should be stored under recommended conditions as per the certificate of analysis.
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| Toxicity/Toxicokinetics |
The toxicity profile of M443 is related to its mechanism as an MRK inhibitor. The compound has been shown to selectively radiosensitize tumor cells without affecting normal brain cells. This selectivity suggests a favorable therapeutic window. Standard toxicological studies would be required for clinical development.
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| References |
[1]. Markowitz D, et al. Pharmacological Inhibition of the Protein Kinase MRK/ZAK Radiosensitizes Medulloblastoma. Mol Cancer Ther. 2016 Aug;15(8):1799-808
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| Additional Infomation |
M443 is a research tool for studying DNA damage response pathways, focusing on the roles of key proteins such as p38 MAPK and Chk2. It is used in cancer research to investigate the potential of combining MRK inhibition with radiation therapy. The compound is also known as EVT-274557.
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| Molecular Formula |
C31H30F3N7O2
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|---|---|
| Molecular Weight |
589.610816478729
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| Exact Mass |
589.241
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| CAS # |
1820684-31-8
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| PubChem CID |
92042851
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
5
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
43
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| Complexity |
984
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=C(C=C(C=C1)C(=O)NC2=CC(=CC(=C2)C(F)(F)F)N3C=C(N=C3)C)NC4=NC=CC(=N4)C5CCCN(C5)C(=O)C=C
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| InChi Key |
QAQFNUYJUCJMKF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C31H30F3N7O2/c1-4-28(42)40-11-5-6-22(17-40)26-9-10-35-30(38-26)39-27-12-21(8-7-19(27)2)29(43)37-24-13-23(31(32,33)34)14-25(15-24)41-16-20(3)36-18-41/h4,7-10,12-16,18,22H,1,5-6,11,17H2,2-3H3,(H,37,43)(H,35,38,39)
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| Chemical Name |
4-methyl-N-[3-(4-methylimidazol-1-yl)-5-(trifluoromethyl)phenyl]-3-[[4-(1-prop-2-enoylpiperidin-3-yl)pyrimidin-2-yl]amino]benzamide
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| Synonyms |
M 443; M-443; M443
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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) |
DMSO : ~55 mg/mL (~93.28 mM)
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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 | 1.6960 mL | 8.4802 mL | 16.9604 mL | |
| 5 mM | 0.3392 mL | 1.6960 mL | 3.3921 mL | |
| 10 mM | 0.1696 mL | 0.8480 mL | 1.6960 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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