ML240 is a novel potent p97 inhibitor which inhibits p97 ATPase with an IC50 value of 100 nM. ML240 inhibited degradation of a p97-dependent but not a p97-independent proteasome substrate in a dual-reporter cell line. ML240 also impaired the endoplasmic-reticulum-associated degradation (ERAD) pathway. The AAA ATPase p97 is a critical factor in maintaining protein homeostasis in eukaryotic cells, through its roles in promoting degradation of ubiquinated proteins by the proteasome and in maturation of autophagosomes. ML241 has the potential for treating cancer.
Physicochemical Properties
| Molecular Formula | C23H20N6O | |
| Molecular Weight | 396.45 | |
| Exact Mass | 396.17 | |
| Elemental Analysis | C, 69.68; H, 5.09; N, 21.20; O, 4.04 | |
| CAS # | 1346527-98-7 | |
| Related CAS # |
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| PubChem CID | 49830258 | |
| Appearance | Light yellow to yellow solid powder | |
| Density | 1.4±0.1 g/cm3 | |
| Boiling Point | 696.6±65.0 °C at 760 mmHg | |
| Flash Point | 375.1±34.3 °C | |
| Vapour Pressure | 0.0±2.2 mmHg at 25°C | |
| Index of Refraction | 1.718 | |
| LogP | 3.75 | |
| Hydrogen Bond Donor Count | 2 | |
| Hydrogen Bond Acceptor Count | 6 | |
| Rotatable Bond Count | 5 | |
| Heavy Atom Count | 30 | |
| Complexity | 558 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | O(C([H])([H])[H])C1=C([H])C([H])=C([H])C2=C1N=C(N=C2N([H])C([H])([H])C1C([H])=C([H])C([H])=C([H])C=1[H])N1C(N([H])[H])=NC2=C([H])C([H])=C([H])C([H])=C12 |
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| InChi Key | NHAMBLRUUJAFOY-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C23H20N6O/c1-30-19-13-7-10-16-20(19)27-23(28-21(16)25-14-15-8-3-2-4-9-15)29-18-12-6-5-11-17(18)26-22(29)24/h2-13H,14H2,1H3,(H2,24,26)(H,25,27,28) | |
| Chemical Name | 2-(2-aminobenzimidazol-1-yl)-N-benzyl-8-methoxyquinazolin-4-amine | |
| Synonyms | ML240; ML 240; ML-240 | |
| 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 |
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| 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 | p97 ATPase ( IC50 = 0.11 μM ); p97 ATPase ( Ki = 0.22 μMM ) |
| ln Vitro | ML240 has an IC50 of 100 nM, making it a strong p97 inhibitor. In the UbG76V-GFP stabilization assay, ML240 exhibits activity (IC50, 0.9 μM). ML240 has a 0.22 μM Ki value and inhibits p97 in a competitive manner in relation to ATP. Furthermore, when tested at 20 μM, ML240 inhibits by more than 50% the labeling of just three protein kinase domains: DNAPK (DNA-dependent protein kinase), JAK1 JH2 (N-terminal pseudokinase domain of JAK1), and PIP5 K3 (phosphoinositide-3 kinase family). Independent of apical caspases 8 and 9, ML240 (1.1, 3.3, 10, or 20 μM) induces executioner caspases 3 and 7 and causes cell death[1].With GI50s of 0.76 and 0.5 μM after treatment for 24 hours, and 0.54 and 0.5 μM after treatment for 72 hours, respectively, ML240 is cytotoxic to HCT15 and SW403 cells[2]. |
| Cell Assay | HeLa cells that are stable in their expression of ODD-luciferase are seeded (5000 cells/well) onto a 96-well white solid bottom plate and allowed to grow for 16 hours. Following a one-hour treatment with DMEM containing MG132 (4 μM), cells are twice washed with 100 μL PBS. In the well, cycloheximide (50 μg/mL), ML240, and DMEM containing 2.5% FBS are added.One of the four 96-well plates that have been prepared is removed from the incubator at each time interval (70, 90, 120, or 150 minutes). Each well holds 50 μL of medium. Luciferin (50 μL of 1 mg/mL in PBS) is added and incubated for 5 minutes at room temperature with 500 rpm shaking. The Synergy HT Microplate Reader uses an integration time of 0.1 ms to determine luminosity intensity[2]. |
| References |
[1]. Structure-activity relationship study reveals ML240 and ML241 as potent and selective inhibitors of p97 ATPase. ChemMedChem, 2013 Feb, 8(2):297-312. [2]. Selective, reversible inhibitors of the AAA ATPase p97. Probe Reports from the NIH Molecular Libraries Program. April 14, 2011. |
| Additional Infomation | ML240 is a member of the class of quinazolines that is quinazoline which is substituted at positions 2, 5 and 8 by 2-amino-1H-benzimidazol-1-yl, benzylnitrilo and methoxy groups, respectively. It is a ATP-competetive inhibitor of AAA ATPase p97, also known as valosin-containing protein (VCP). It has a role as an antineoplastic agent. It is a member of quinazolines, a member of benzimidazoles, a secondary amino compound, an aromatic amine, an aromatic ether and a primary amino compound. |
Solubility Data
| Solubility (In Vitro) |
DMSO : 12.5~79 mg/mL ( 31.5~199.3 mM ) Ethanol : < 1 mg/mL |
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (6.31 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. 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 (6.31 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (6.31 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5224 mL | 12.6119 mL | 25.2239 mL | |
| 5 mM | 0.5045 mL | 2.5224 mL | 5.0448 mL | |
| 10 mM | 0.2522 mL | 1.2612 mL | 2.5224 mL |