Physicochemical Properties
| Molecular Formula | C19H13NO4 |
| Molecular Weight | 319.31 |
| Exact Mass | 319.084 |
| Elemental Analysis | C, 71.47; H, 4.10; N, 4.39; O, 20.04 |
| CAS # | 175026-96-7 |
| Related CAS # | 175026-96-7 |
| PubChem CID | 5522952 |
| Appearance | Yellow to orange solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 581.4±50.0 °C at 760 mmHg |
| Flash Point | 305.4±30.1 °C |
| Vapour Pressure | 0.0±1.6 mmHg at 25°C |
| Index of Refraction | 1.685 |
| LogP | 0.94 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 4 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 24 |
| Complexity | 623 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O=C1C2=C([H])C([H])=C([H])C([H])=C2C2=C(C(=O)OC([H])([H])C([H])([H])[H])C(N([H])C3=C([H])C([H])=C([H])C1=C23)=O |
| InChi Key | UFJGFNHRMPMALC-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C19H13NO4/c1-2-24-19(23)16-15-10-6-3-4-7-11(10)17(21)12-8-5-9-13(14(12)15)20-18(16)22/h3-9H,2H2,1H3,(H,20,22) |
| Chemical Name | ethyl 8,15-dioxo-14-azatetracyclo[7.7.1.02,7.013,17]heptadeca-1(16),2,4,6,9(17),10,12-heptaene-16-carboxylate |
| Synonyms | NQDI 1; NQDI1; NQDI-1 |
| 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 |
| 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 | ASK1 (IC50 = 3 μM) |
| ln Vitro | The selectivity of NQDI-1 is examined in vitro using four serine/threonine protein kinases (protein kinase CK2, c-Jun N-terminal kinase 3, Rho-associated protein kinase 1, and Aurora A), and three tyrosine protein kinases (FGFR1, hHGFR, and endothelial TEK tyrosine kinase, or Tie2). The outcomes demonstrate that NQDI-1 is a specific ASK1 inhibitor. NQDI-1 inhibits the activity of FGFR1 protein kinase (44% residual activity). With a Ki of 500 nM, NQDI-1 blocks ASK1. When compared to the phosphodonor substrate ATP, NQDI-1's inhibition of ASK1 is competitive[1]. |
| ln Vivo | Following a brain injury, 250 nmol NQDI-1 in DMSO is injected intracerebrally. Western blotting is used to analyze the expression of ASK1 in the hypoxia-ischemia (HI), DMSO, NQDI-1, and sham groups.Results show that NQDI-1 significantly inhibits ASK1 expression in the brain cortex when compared to the HI and DMSO groups. Furthermore, immunofluorescence staining shows that NQDI-1 inhibits ASK1 expression in the cortex of the brain. The current study also identifies the expression of ASK1's downstream targets. When compared to the HI and DMSO groups, NQDI-1 significantly lowers the expression levels of p-JNK, p-c-Jun, p53, and caspase 3. In the group that received NQDI-1 treatment, immunofluorescence also revealed low expression of p-JNK in the cerebral cortex[2]. |
| Enzyme Assay | In vitro kinase assay (γ-32P-ATP method) is used to assess the enzyme activity of human protein kinases ASK1, Aurora A, ROCK1, HGFR, FGFR1, Tie2, JNK3, and CK2. Each reaction mixture contains 6 μL of buffer solution (25 mM MOPS, pH 7.2, 2.5 mM EGTA, 2.5 mM EDTA, 0.5 mM DTT, 0.25 mg/mL BSA, 20 mM β-glycerophosphate), 3 μL of substrate solution (MBP, Long S6 kinase substrate peptide, KKKSPGEYVNIEFG, IGF-IRtide (12-527), TK substrate 2, JNK3tide, or RRRDDDSDDD for each kinase, respectively) (5.0 μg/μL), 0.3 μL of enzyme (protein kinase catalytic subunit, 0.1 μg/μL≈32 mU/μL), and 10.25 μL of H2O. At room temperature, the reaction mixture (19 μL total volume) is swiftly added to 1.5 mL tubes. Stock solutions of inhibitors, like NQDI-1, are made in DMSO with a 1 mM inhibitor concentration. In the reaction, the amount of DMSO is not greater than 3%. After that, each tube receives 1 L of an inhibitor solution in DMSO, which is pipetted into each tube and mixed. A separate ATP solution is made. 0.05 mCi of γ-[P32]ATP is extracted for each sample (specific activity of 100 μCi/M). 100 μM is the combined concentration of labeled and unlabeled ATP. The 150 μM ATP, 30 mM MgCl2, 15 mM MOPS, pH 7.2 ATP solution is added to start the reaction. The reaction lasts 20 minutes at 30 °C. 20 μL of 0.5 M orthophosphoric acid are added to stop the reaction. Following that, the reaction mixture is loaded onto the cellulose phosphate paper 20 mm filter disks. At room temperature, filters are washed three times with 0.075 M orthophosphoric acid, then dried. By using a Tri-Carb 2800-TR liquid scintillation analyzer, dried filters are counted in order to detect products. A positive control is then performed by substituting 1 μL of DMSO for the inhibitor stock solution in the reaction volume[1]. |
| References |
[1]. Identification of 3H-naphtho[1,2,3-de]quinoline-2,7-diones as inhibitors of apoptosis signal-regulating kinase 1 (ASK1). J Med Chem. 2011 Apr 28;54(8):2680-6. [2]. NQDI-1, an inhibitor of ASK1 attenuates acute perinatal hypoxic-ischemic cerebral injury by modulating cell death. Mol Med Rep. 2016 Jun;13(6):4585-92. |
Solubility Data
| Solubility (In Vitro) | DMSO: ~21 mg/mL (~65.8 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: 0.71 mg/mL (2.22 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 7.1 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: 0.71 mg/mL (2.22 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 7.1 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: 5%dmso+40%PEG300+5%tween-80+ddH2O: 0.5mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1318 mL | 15.6588 mL | 31.3175 mL | |
| 5 mM | 0.6264 mL | 3.1318 mL | 6.2635 mL | |
| 10 mM | 0.3132 mL | 1.5659 mL | 3.1318 mL |