Physicochemical Properties
| Molecular Formula | C22H27CLN8O3S |
| Molecular Weight | 519.019581079483 |
| CAS # | 2244775-31-1 |
| PubChem CID | 138454747 |
| Appearance | White to off-white solid powder |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 11 |
| Rotatable Bond Count | 8 |
| Heavy Atom Count | 35 |
| Complexity | 766 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | ClC1C=NC(=NC=1NC1C=CC=CC=1NS(C)(=O)=O)NC1=C(N=C(C=C1)N1CCN(C)CC1)OC |
| 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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 |
ALK^{WT} (IC₅₀ = 2.1 nM), ROS1^{WT} (IC₅₀ = 2.3 nM), ALK^{L1196M} (IC₅₀ = 1.3 nM), ALK^{G1202R} (IC₅₀ = 3.9 nM) [1] |
| ln Vitro |
F-1 exhibited significant cytotoxicity on ALK-addicted Karpas299, H2228, and Ba/F3 cell expressing G1202R mutant, as well as ROS1-positive HCC78 cell with IC50 values ranging from 10 nM to 43 nM. Notably, F-1 was capable of suppressing phospho-ALK and its relative downstream signaling pathways, and eventually, inducing cell apoptosis in a dose-dependent manner in Karpas-299 cell. Together, F-1 is validated as a promising ALK/ROS1 dual inhibitor great potential for G1202R ALK mutation cancers. 1. Enzymatic activity: F-1 exhibited potent inhibitory effects on ALK^{WT}, ROS1^{WT}, ALK^{L1196M} and ALK^{G1202R} kinases with IC₅₀ values of 2.1 nM, 2.3 nM, 1.3 nM and 3.9 nM, respectively, and its inhibitory activity was superior to that of crizotinib and ceritinib [1] 2. Cytotoxicity: F-1 showed significant cytotoxicity against ALK-addicted Karpas299, H2228, Ba/F3 cells expressing G1202R mutant, as well as ROS1-positive HCC78 cells, with IC₅₀ values ranging from 10 nM to 43 nM [1] 3. Cellular signaling and apoptosis: In Karpas-299 cells, F-1 suppressed phospho-ALK and its downstream signaling pathways in a dose-dependent manner, and ultimately induced cell apoptosis in a dose-dependent manner [1] |
| Enzyme Assay |
1. A series of novel 2,4-diarylaminopyrimidine analogues incorporating 2-alkoxy-6-alicyclic aminopyridinyl motifs were designed and synthesized to identify ALK/ROS1 dual inhibitors against drug-resistant mutants. Enzymatic assays were conducted to evaluate the inhibitory activity of the synthesized compounds against ALK^{WT}, ROS1^{WT}, ALK^{L1196M} and ALK^{G1202R} kinases, and the IC₅₀ values of F-1 for these kinases were determined and compared with those of crizotinib and ceritinib [1] |
| Cell Assay |
1. Cytotoxicity assay: ALK-addicted cell lines (Karpas299, H2228), Ba/F3 cells expressing ALK^{G1202R} mutant and ROS1-positive HCC78 cells were used to assess the cytotoxicity of F-1. The cells were treated with different concentrations of F-1, and the cell viability was detected to calculate the IC₅₀ values of F-1 for these cell lines [1] 2. Western blot and apoptosis assay: Karpas-299 cells were treated with F-1 at different doses. Western blot was performed to detect the expression level of phospho-ALK and its downstream signaling molecules, and apoptosis assays were carried out to evaluate the apoptotic effect of F-1 on Karpas-299 cells and the dose-dependent relationship [1] |
| References |
[1]. Dual potent ALK and ROS1 inhibitors combating drug-resistant mutants: Synthesis and biological evaluation of aminopyridine-containing diarylaminopyrimidine derivatives. Eur J Med Chem. 2018 Sep 6;158:322-333. |
| Additional Infomation |
1. F-1 was identified from a series of novel 2,4-diarylaminopyrimidine analogues designed and synthesized by incorporating 2-alkoxy-6-alicyclic aminopyridinyl motifs, which were developed to find ALK and ROS1 dual inhibitors conferring resistance to ALK secondary mutations (especially the 'gatekeeper' L1196M and ceritinib-resistant G1202R mutations) [1] 2. F-1 is validated as a promising ALK/ROS1 dual inhibitor with great potential for the treatment of cancers harboring the G1202R ALK mutation [1] |
Solubility Data
| Solubility (In Vitro) | DMSO : ~125 mg/mL (~240.84 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.01 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 20.8 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: 2.08 mg/mL (4.01 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 20.8 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: ≥ 2.08 mg/mL (4.01 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 20.8 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 | 1.9267 mL | 9.6335 mL | 19.2671 mL | |
| 5 mM | 0.3853 mL | 1.9267 mL | 3.8534 mL | |
| 10 mM | 0.1927 mL | 0.9634 mL | 1.9267 mL |