| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
Aurora-A (IC50 = 0.031 μM)
Phthalazinone pyrazole (1 and 10 μM; 30 hours) increases the ability of HLCs to proliferate[2]. Phthalazinone pyrazole (1, 10, and 100 μM; 5 days) increases the morphological changes of the liver in differentiated HLCs without causing cytotoxicity[2]. Phthalazinone pyrazole (1 and 10 μM; 5 and 17 days) inhibits the AKT signaling pathway through an off-target effect and concurrent upregulation of HNF4α, rather than directly inhibiting Aurora-A, which suppresses the EMT and induces maturation of HLCs. Western blot and qPCR are used to confirm the outcome[2]. Phthalazinone pyrazoles are potent and selective inhibitors of Aurora-A kinase. Representative compounds such as 7 (IC₅₀ = 0.031 μM), 8 (0.029 μM), 9 (0.023 μM), 10 (0.024 μM), 11 (0.24 μM), 12 (0.093 μM), 13 (0.037 μM), 16 (0.065 μM), 17 (0.049 μM), 18 (0.025 μM), 19 (0.014 μM), 20 (0.056 μM), 21 (0.032 μM), 22 (0.036 μM), 23 (0.093 μM), 24 (0.27 μM), 25 (0.017 μM), 26 (0.109 μM), 27 (0.067 μM), 28 (0.1 μM), 29 (0.72 μM), 30 (0.071 μM), 31 (0.12 μM), 32 (0.27 μM), 33 (0.035 μM), 34 (0.115 μM), 35 (0.066 μM), and 36 (0.55 μM) showed >1000-fold selectivity over Aurora-B (Aurora-B IC₅₀ typically >100 μM for most compounds). [1] |
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| ln Vitro |
Phthalazinone pyrazole (1 and 10 μM; 30 hours) increases the ability of HLCs to proliferate[2].
Phthalazinone pyrazole (1, 10, and 100 μM; 5 days) increases the morphological changes of the liver in differentiated HLCs without causing cytotoxicity[2]. Phthalazinone pyrazole (1 and 10 μM; 5 and 17 days) inhibits the AKT signaling pathway through an off-target effect and concurrent upregulation of HNF4α, rather than directly inhibiting Aurora-A, which suppresses the EMT and induces maturation of HLCs. Western blot and qPCR are used to confirm the outcome[2]. Compounds in this series exhibited antiproliferative activity in multiple cancer cell lines, including HCT116, Colo205, and MCF7. For example, compound 7 showed IC₅₀ values of 7.8 μM in HCT116, 2.9 μM in Colo205, and 1.6 μM in MCF7. Compound 33 showed IC₅₀ values of 0.4 μM in HCT116, 1.7 μM in Colo205, and 1.6 μM in MCF7. Several compounds induced a phenotypic Aurora-A inhibition in MCF7 cells, leading to poorly formed centrosomes and arrest at the G2/M phase. [1] Compounds in this series exhibited antiproliferative activity in multiple cancer cell lines, including HCT116, Colo205, and MCF7. For example, compound 7 showed IC₅₀ values of 7.8 μM in HCT116, 2.9 μM in Colo205, and 1.6 μM in MCF7. Compound 33 showed IC₅₀ values of 0.4 μM in HCT116, 1.7 μM in Colo205, and 1.6 μM in MCF7. Several compounds induced a phenotypic Aurora-A inhibition in MCF7 cells, leading to poorly formed centrosomes and arrest at the G2/M phase. [1] |
| ln Vivo |
Compounds 16 and 33 showed plasma concentrations above the cellular IC₅₀ after oral administration at 30 mg/kg, with Cmax values of 13 μM and 25 μM, respectively. Their oral bioavailability was substantially improved compared to VX-680. [1]
Compounds 16 and 33 showed plasma concentrations above the cellular IC₅₀ after oral administration at 30 mg/kg, with Cmax values of 13 μM and 25 μM, respectively. Their oral bioavailability was substantially improved compared to VX-680. [1] |
| Enzyme Assay |
Aurora kinase activity was measured using a biochemical kinase assay. Recombinant Aurora-A or Aurora-B kinase was incubated with test compounds and 30 μM ATP in kinase buffer supplemented with blocking agent. The reaction was stopped after 40 minutes by adding EDTA. Phosphorylation of histone H3 (Ser10) was detected using an anti-phospho-histone H3 antibody and a secondary HRP-conjugated antibody, followed by colorimetric development with TMB. Absorbance was read, and IC₅₀ values were calculated using nonlinear curve fitting. All data were averaged from triplicate experiments. [1]
Aurora kinase activity was measured using a biochemical kinase assay. Recombinant Aurora-A or Aurora-B kinase was incubated with test compounds and 30 μM ATP in kinase buffer supplemented with blocking agent. The reaction was stopped after 40 minutes by adding EDTA. Phosphorylation of histone H3 (Ser10) was detected using an anti-phospho-histone H3 antibody and a secondary HRP-conjugated antibody, followed by colorimetric development with TMB. Absorbance was read, and IC₅₀ values were calculated using nonlinear curve fitting. All data were averaged from triplicate experiments. [1] |
| Cell Assay |
Cell viability was assessed using the CellTiter-Glo assay in HCT116 cells. Cells were seeded in 384-well plates at 1000 cells per well and treated with test compounds at concentrations ranging from 30 μM to 0.0015 μM (10-point, 1:3 dilution). After 5 days, CellTiter-Glo reagent was added, and luminescence was measured after cell lysis. Data were averaged from triplicate experiments. [1]
Cell viability was assessed using the CellTiter-Glo assay in HCT116 cells. Cells were seeded in 384-well plates at 1000 cells per well and treated with test compounds at concentrations ranging from 30 μM to 0.0015 μM (10-point, 1:3 dilution). After 5 days, CellTiter-Glo reagent was added, and luminescence was measured after cell lysis. Data were averaged from triplicate experiments. [1] |
| Animal Protocol |
Pharmacokinetic studies were conducted in mice. Compounds were administered orally at 30 mg/kg. Plasma concentrations were measured over time to determine Cmax, half-life (T₁/₂), and oral bioavailability. Specific formulations and vehicle details were not described in the experimental section. [1]
Pharmacokinetic studies were conducted in mice. Compounds were administered orally at 30 mg/kg. Plasma concentrations were measured over time to determine Cmax, half-life (T₁/₂), and oral bioavailability. Specific formulations and vehicle details were not described in the experimental section. [1] |
| ADME/Pharmacokinetics |
Some compounds exhibited moderate to high stability in microsomes and hepatocytes. For example, compound 7 had a hepatic clearance (CLhep) of 2.3 μL/min/10⁶ cells (rat) and a microsomal clearance (CLmic) of 0 μL/min/mg protein (human) and 15.7 μL/min/mg protein (mouse), respectively. Compound 16 had CLmic of 7.4 μL/min/mg protein (human) and 28 μL/min/mg protein (mouse), respectively. Compound 33 had a half-life of 2.4 hours and a Cmax of 25 μM after oral administration of 30 mg/kg. Its oral bioavailability was significantly improved compared to VX-680. [1] For example, compound 7 had a hepatic clearance (CLhep) of 2.3 μL/min/10⁶ cells (rat) and a microsomal clearance (CLmic) of 0 μL/min/mg protein (human) and 15.7 μL/min/mg protein (mouse). Compound 16 had a Cmax of 7.4 μL/min/mg protein (human) and 28 μL/min/mg protein (mouse). Compound 33 had a half-life of 2.4 hours and a Cmax of 25 μM after oral administration of 30 mg/kg. Its oral bioavailability was significantly improved compared to VX-680. [1]
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| Toxicity/Toxicokinetics |
At the tested concentrations (e.g., >50 μM for CYP3A4, 2C9, and 2D6), no significant cytochrome P450 inhibition or hERG channel inhibition was observed in compounds 7, 16, and 33. Compound 7 showed a 20% hERG inhibition rate at 10 μM. [1]
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| References |
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| Additional Infomation |
Phthalasinonepyrazole is a novel class of Aurora-A inhibitors with extremely high selectivity (>1000-fold) for Aurora-B. They exhibit good oral bioavailability and plasma exposure, making them suitable for pharmacodynamic studies. The crystal structure of compound 7 bound to Aurora-A reveals a key hydrogen bond with the hinge region, explaining its selectivity for Aurora-B as due to steric hindrance to glutamate residues in Aurora-B. [1]
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| Molecular Formula |
C18H15N5O
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|---|---|
| Molecular Weight |
317.344602823257
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| Exact Mass |
317.127
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| Elemental Analysis |
C, 68.13; H, 4.76; N, 22.07; O, 5.04
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| CAS # |
880487-62-7
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| Related CAS # |
880487-62-7;
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| PubChem CID |
11652621
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| Appearance |
Brown to gray solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
557.7±52.0 °C at 760 mmHg
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| Flash Point |
291.1±30.7 °C
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| Vapour Pressure |
0.0±1.5 mmHg at 25°C
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| Index of Refraction |
1.716
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| LogP |
3.03
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
24
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| Complexity |
502
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C1N(C2C=CC=CC=2)N=C(NC2C=C(C)NN=2)C2C1=CC=CC=2
|
| InChi Key |
DSDIWWSXOOXFSI-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H15N5O/c1-12-11-16(21-20-12)19-17-14-9-5-6-10-15(14)18(24)23(22-17)13-7-3-2-4-8-13/h2-11H,1H3,(H2,19,20,21,22)
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| Chemical Name |
4-[(5-methyl-1H-pyrazol-3-yl)amino]-2-phenylphthalazin-1-one
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| Synonyms |
Phthalazinone pyrazole
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO: ≥ 11.11 mg/mL (~35.0 mM)
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1512 mL | 15.7560 mL | 31.5119 mL | |
| 5 mM | 0.6302 mL | 3.1512 mL | 6.3024 mL | |
| 10 mM | 0.3151 mL | 1.5756 mL | 3.1512 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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