ML365 is a potent, novel and selective small molecule inhibitor of the TASK1(KCNK3, a two-pore domain potassium channel). ML365 blocks TASK1 channels in both the thallium influx fluorescent assay (IC50 = 4 nM) and an automated electrophysiology assay (IC50 = 16 nM). Based on potency differences, it possesses more than 60-fold selectivity for inhibition of TASK1 over a closely-related, two-pore domain potassium channel, TASK3. ML365 displays little or no inhibition at 30 μM of more distantly related potassium channels, Kir2.1, potassium voltage-gated channel, KQT-like subfamily, member 2 (KCNQ2), and human ether-a go-go-related gene (hERG). Based on these criteria, ML365 is a best-in-class probe and is a useful pharmacological probe for in vitro studies of TASK1 function and in further studies aimed at developing therapeutic intervention.
Physicochemical Properties
| Molecular Formula | C22H20N2O3 | |
| Molecular Weight | 360.405805587769 | |
| Exact Mass | 360.147 | |
| Elemental Analysis | C, 73.32; H, 5.59; N, 7.77; O, 13.32 | |
| CAS # | 947914-18-3 | |
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| PubChem CID | 46785920 | |
| Appearance | White to off-white solid powder | |
| Density | 1.253±0.06 g/cm3 (20 °C, 760 mmHg) | |
| Boiling Point | 429.9±40.0 °C (760 mmHg) | |
| LogP | 4.654 | |
| Hydrogen Bond Donor Count | 2 | |
| Hydrogen Bond Acceptor Count | 3 | |
| Rotatable Bond Count | 5 | |
| Heavy Atom Count | 27 | |
| Complexity | 510 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | O=C(C1C=C(C)C=CC=1)NC1C=C(NC(C2C(OC)=CC=CC=2)=O)C=CC=1 |
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| InChi Key | UTAJHKSGYJSZBR-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C22H20N2O3/c1-15-7-5-8-16(13-15)21(25)23-17-9-6-10-18(14-17)24-22(26)19-11-3-4-12-20(19)27-2/h3-14H,1-2H3,(H,23,25)(H,24,26) | |
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| 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 |
ML365 is a selective inhibitor of the KCNK3/TASK1 two - pore potassium channel, with an IC50 of 4 nM in the thallium influx fluorescent assay and 16 nM in the automated electrophysiology assay. It has more than 60 - fold selectivity for inhibiting TASK1 over TASK3 [1] |
| ln Vitro |
ML365 blocks TASK1 channels in a dose - dependent manner. In the thallium influx fluorescent assay, it can inhibit the influx of thallium through TASK1 channels with an IC50 of 4 nM. In an automated electrophysiology assay, the IC50 for blocking TASK1 channels is 16 nM. At a concentration of 30 μM, it has little or no inhibitory effect on Kir2.1, KCNQ2, and hERG potassium channels [1] In automated electrophysiological studies (IC50 = 16 nM) and thallium influx fluorescence assays (IC50 = 4 nM), ML365 inhibits TASK1 channels [1]. At 30 μM or no inhibitory impact, ML365 has minimal activity against the potassium voltage-gated channel, KQT-like subfamily, member 2 (KCNQ2), human ether-a go-go related gene (hERG), and the distantly related potassium channel Kir2.1 [1]. ML365 does not show acute toxicity in cell-based studies at doses as high as 30 μM [1]. |
| Enzyme Assay |
For the thallium influx fluorescent assay, TASK1 - expressing cells are incubated with thallium ions and a fluorescent dye. Then, different concentrations of ML365 (ranging from low to high) are added, and the fluorescence intensity is measured. The fluorescence intensity is related to the influx of thallium through TASK1 channels, and the IC50 of ML365 can be calculated according to the dose - response curve. For the automated electrophysiology assay, the patch - clamp technique is used to record the current of TASK1 channels. Cells expressing TASK1 channels are clamped at a certain potential, and different concentrations of ML365 are added, and the change of channel current is observed to determine the inhibitory effect of ML365 on TASK1 channels and calculate the IC50 value [1] |
| References |
[1]. ML365: Development of Bis-Amides as Selective Inhibitors of the KCNK3/TASK1 Two Pore Potassium Channel. Probe Reports from the NIH Molecular Libraries Program [Internet]. https://pubmed.ncbi.nlm.nih.gov/24479195/ |
| Additional Infomation |
Two - pore domain potassium channels play important roles in regulating cell membrane potential, and TASK1 is one of them. ML365 was obtained by high - throughput fluorescent screening of the Molecular Libraries Small Molecule Repository (MLSMR) library and structure - activity relationship (SAR) analysis of active compounds. It is a novel selective small - molecule inhibitor of TASK1, which can be used as a useful pharmacological probe for in vitro studies of TASK1 function and is helpful for further research on therapeutic interventions [1] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3 mg/mL (8.32 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 30.0 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: ≥ 3 mg/mL (8.32 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 30.0 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 | 2.7746 mL | 13.8731 mL | 27.7462 mL | |
| 5 mM | 0.5549 mL | 2.7746 mL | 5.5492 mL | |
| 10 mM | 0.2775 mL | 1.3873 mL | 2.7746 mL |