ZM 447439

Catalog No.S1103 Batch:S110303

Technical Data

Formula

C₂₉H₃₁N₅O₄

Molecular Weight

513.59

CAS No.

331771-20-1

Solubility (25°C)*

In vitro

DMSO

50 mg/mL (97.35 mM)

Water

Insoluble

Ethanol

Insoluble

In vivo (Add solvents to the product individually and in order)

Clear solution

30%PEG400 1 0.5%Tween80 2 5%propylene glycol

30.0mg/ml

Taking the 1 mL working solution as an example, add 300 μL of 100 mg/ml clarified PEG400 stock solution to 5 μL of Tween80, mix evenly to clarify it; add 50 μL Propylene glycol to the above system, mix evenly to clarify it; then continue to add 645 μL ddH2O to adjust the volume. to 1 mL. The mixed solution should be used immediately for optimal results.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

ZM 447439 is a selective and ATP-competitive inhibitor for Aurora A and Aurora B with IC50 of 110 nM and 130 nM, respectively. It is more than 8-fold selective for Aurora A/B than MEK1, Src, Lck and has little effect against CDK1/2/4, Plk1, Chk1, etc.

Targets

Aurora A ^[1] (Cell-free assay)	Aurora B ^[1] (Cell-free assay)	LCK ^[1] (Cell-free assay)	Src ^[1] (Cell-free assay)	MEK1 ^[1] (Cell-free assay)
110 nM	130 nM	880 nM	1.03 μM	1.79 μM

In vitro

In vitro, ZM-447439 selectively inhibits recombinant human Aurora A and B with IC50 values of 110 and 130 nM, respectively, while other protein kinases of diverse structural types including the mitotic kinases CDK1 and PLK1 are inhibited with IC50 values >10 μM. ^[1] Aurora kinase inhibitor, ZM-447439 time- and dose-dependently inhibits the growth of all three cell lines with IC50 values of 3 μM (BON), 0.9 μM (QGP-1) and 3 μM (MIP-101) after 72 hours of continuous exposure. In addition, ZM-447439 potently induces cell apoptosis by promoting DNA fragmentation and caspase 3 and 7 activation, and arrests GEP-NET cells in the G₀ /G₁and G₂/M phase of the cell cycle. ^[2] In mouse embryo, inhibition of Aurora kinase activity by ZM-447439 results in abnormalities during mitosis by regulating the phosphorylation of histone H3 serine 10 (H3S10Ph) from G2 to metaphase with different perturbations in each embryonic cycle. ^[3] A recent study shows that ZM-447439 exhibits growth inhibitory and proapoptotic effect on cervical cancer SiHa cells, and enhances the chemosensitivity to cisplatin. ^[4]

Features

An Aurora selective ATP-competitive inhibitor.

Protocol (from reference)

Kinase Assay:^{^[1]}	In vitro kinase assays Recombinant Aurora A and B are expressed as NH₂-terminal His₆-tagged fusion proteins using a baculovirus expression system. Aurora A is purified by affinity chromatography using Ni-NTA agarose, and Aurora B is purified by ion exchange chromatography using CM Sepharose Fast Flow. 1 ng purified recombinant enzyme is added to a reaction cocktail containing 25 mM Tris-HCl, pH 7.5, 12.5 mM KCl, 2.5 mM NaF, 0.6 mM DTT, 6.25 mM MnCl₂, 10 μM peptide substrate, 10 μM for Aurora A or 5 μM ATP for Aurora B, and 0.2 μCi γ-[³³P]ATP (specific activity ≥2,500 Ci/mmol), and is then incubated at RT for 60 minutes. Reactions are stopped by addition of 20% phosphoric acid, and the products are captured on P30 nitrocellulose filters and assayed for incorporation of ³³P with a Betaplate^TM counter. No enzyme and no compound control values are used to determine the concentration of ZM447439, which gave 50% inhibition of enzyme activity. Further details are available on request from Nicholas Keen.
Cell Assay:^{^[2]}	Cell lines BON, QGP-1 and MIP-101 cells Concentrations 0-5 μM Incubation Time 72 hours Method Cell number is evaluated by crystal violet staining. In brief, cells in 96-well plates are fixed with 1% glutaraldehyde. Then cells are stained with 0.1% crystal violet. The unbound dye is removed by washing with water. Bound crystal violet is solubilized with 0.2% Triton X-100. Light extinction which increases linearly with the cell number is analyzed at 570 nm using an ELISA reader.

Kinase Assay:^{^[1]}

In vitro kinase assays
Recombinant Aurora A and B are expressed as NH₂-terminal His₆-tagged fusion proteins using a baculovirus expression system. Aurora A is purified by affinity chromatography using Ni-NTA agarose, and Aurora B is purified by ion exchange chromatography using CM Sepharose Fast Flow. 1 ng purified recombinant enzyme is added to a reaction cocktail containing 25 mM Tris-HCl, pH 7.5, 12.5 mM KCl, 2.5 mM NaF, 0.6 mM DTT, 6.25 mM MnCl₂, 10 μM peptide substrate, 10 μM for Aurora A or 5 μM ATP for Aurora B, and 0.2 μCi γ-[³³P]ATP (specific activity ≥2,500 Ci/mmol), and is then incubated at RT for 60 minutes. Reactions are stopped by addition of 20% phosphoric acid, and the products are captured on P30 nitrocellulose filters and assayed for incorporation of ³³P with a Betaplate^TM counter. No enzyme and no compound control values are used to determine the concentration of ZM447439, which gave 50% inhibition of enzyme activity. Further details are available on request from Nicholas Keen.

Cell Assay:^{^[2]}

Cell lines
BON, QGP-1 and MIP-101 cells
Concentrations
0-5 μM
Incubation Time
72 hours
Method
Cell number is evaluated by crystal violet staining. In brief, cells in 96-well plates are fixed with 1% glutaraldehyde. Then cells are stained with 0.1% crystal violet. The unbound dye is removed by washing with water. Bound crystal violet is solubilized with 0.2% Triton X-100. Light extinction which increases linearly with the cell number is analyzed at 570 nm using an ELISA reader.

References

[4] Zhang L, et al. J Obstet Gynaecol Res. 2011, 37(6), 591-600.

+ Expand

Customer Product Validation

: Data from [Mol Biol Cell, 2011, 22, 4227-35]

: , , Dr. Yuanhong Chen of University of Nebraska

: , , Dr. Zhang of Tianjin Medical University

Selleck's ZM 447439 has been cited by 69 publications

Detection of senescence using machine learning algorithms based on nuclear features [ Nat Commun, 2024, 15(1):1041]	PubMed: 38310113
Human Endonuclease ANKLE1 Localizes at the Midbody and Processes Chromatin Bridges to Prevent DNA Damage and cGAS-STING Activation [ Adv Sci (Weinh), 2023, 10(12):e2204388]	PubMed: 36825683
Extra centrosomes induce PIDD1-mediated inflammation and immunosurveillance [ EMBO J, 2023, e113510.]	PubMed: 37530438
RIF1 suppresses the formation of single-stranded ultrafine anaphase bridges via protein phosphatase 1 [ Cell Rep, 2023, 42(2):112032]	PubMed: 36719798
Aberrant metabolite trafficking and fuel sensitivity in human pluripotent stem cell-derived islets [ Cell Rep, 2023, 42(8):112970]	PubMed: 37556323
DELs enable the development of BRET probes for target engagement studies in cells [ Cell Chem Biol, 2023, 30(8):987-998.e24]	PubMed: 37490918
PLK1 promotes the mitotic surveillance pathway by controlling cytosolic 53BP1 availability [ EMBO Rep, 2023, e57234.]	PubMed: 37888778
Mitotic DNA damage promotes chromokinesin-mediated missegregation of polar chromosomes in cancer cells [ Mol Biol Cell, 2023, 34(5):ar47]	PubMed: 36989031
Nuclear chromosome locations dictate segregation error frequencies [ Nature, 2022, 607(7919):604-609]	PubMed: 35831506
Pancreas agenesis mutations disrupt a lead enhancer controlling a developmental enhancer cluster [ Dev Cell, 2022, 57(16):1922-1936.e9]	PubMed: 35998583

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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