GNE-220 HCl is a novel, potent and selective inhibitor of MAP4K4 with an IC50 of 7 nM. Cell migration is a stepwise process that coordinates multiple molecular machineries. Using in vitro angiogenesis screens with short interfering RNA and chemical inhibitors, we define here a MAP4K4-moesin-talin-β1-integrin molecular pathway that promotes efficient plasma membrane retraction during endothelial cell migration. Loss of MAP4K4 decreased membrane dynamics, slowed endothelial cell migration, and impaired angiogenesis in vitro and in vivo. In migrating endothelial cells, MAP4K4 phosphorylates moesin in retracting membranes at sites of focal adhesion disassembly.
Physicochemical Properties
| Molecular Formula | C25H27CLN8 |
| Molecular Weight | 474.988482713699 |
| Exact Mass | 474.204 |
| CAS # | 2448286-21-1 |
| Related CAS # | GNE 220;1199590-75-4 |
| PubChem CID | 134828260 |
| Appearance | Light yellow to yellow solid powder |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 6 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 34 |
| Complexity | 661 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | Cl.N1(C2C=CC(C3=CN=C4C(=C3)C3=C(C(C)=NC(C5C=NN(C)C=5)=N3)N4)=CC=2)CCN(C)CC1 |
| InChi Key | VBVBAEHNBOTZQV-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C25H26N8.ClH/c1-16-22-23(30-24(28-16)19-14-27-32(3)15-19)21-12-18(13-26-25(21)29-22)17-4-6-20(7-5-17)33-10-8-31(2)9-11-33;/h4-7,12-15H,8-11H2,1-3H3,(H,26,29);1H |
| Chemical Name | 6-methyl-12-[4-(4-methylpiperazin-1-yl)phenyl]-4-(1-methylpyrazol-4-yl)-3,5,8,10-tetrazatricyclo[7.4.0.02,7]trideca-1(9),2,4,6,10,12-hexaene;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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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
| ln Vitro | GNE-220 also has the ability to inhibit a number of other kinases; its IC50 values for MINK (MAP4K6), DMPK, and KHS1 (MAP4K5) are 9 nM, 476 nM, and 1.1 μM, respectively. The sprouting morphology of human umbilical vein endothelial cells (HUVECs) is altered by GNE-220. In a dose-dependent manner, GNE-220 also decreased pERM+ retractile fibers. Additionally, GNE-220 increases the number of active INTβ1+ focal adhesions (FA) in a dose-dependent manner. [1] |
| References |
[1]. MAP4K4 regulates integrin-FERM binding to control endothelial cell motility. Nature. 2015 Mar 26;519(7544):425-30. |
Solubility Data
| Solubility (In Vitro) |
DMSO : ~5.4 mg/mL (~11.37 mM) H2O : ~5 mg/mL (~10.53 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 0.54 mg/mL (1.14 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 5.4 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: ≥ 0.54 mg/mL (1.14 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 5.4 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: ≥ 0.54 mg/mL (1.14 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 5.4 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 11.11 mg/mL (23.39 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C). (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1053 mL | 10.5265 mL | 21.0531 mL | |
| 5 mM | 0.4211 mL | 2.1053 mL | 4.2106 mL | |
| 10 mM | 0.2105 mL | 1.0527 mL | 2.1053 mL |