| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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Purity: ≥98%
ZM 447439 (ZM-447439) is a novel, potent, selective and ATP-competitive inhibitor of Aurora A and Aurora B with potential antitumor activity. It inhibits Aurora A and Aurora B with IC50s of 110 nM and 130 nM, respectively. It shows 8-fold higher selectivity for Aurora A/B over MEK1, Src, Lck and has little effect against CDK1/2/4, Plk1, Chk1, etc. Being specifically for Aurora kinases, ZM 447439 barely inhibits the majority of other protein kinases (IC50 > 10 μM), such as CDK1/2/4, IKK1/2, PLK1, CHK1, cFLT2, KDR2, FAK and Zap-70, except for MEK1, SRC and LCK (IC50 values of 1.79 μM, 1.03 and 0.88 μM respectively.
| Targets |
Aurora A (IC50 = 110 nM); Aurora B (IC50 = 130 nM)
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| ln Vitro |
ZM-447439-treated cells undergo interphase, properly enter mitosis, and form bipolar spindles. On the other hand, cytokinesis, segregation, and chromosomal alignment all fail. ZM-447439 suppresses the phosphorylation of histone H3 during mitosis and limits cell division. Chromosome alignment and segregation are inhibited by ZM-447439. Function of spindle checkpoint is compromised by ZM-447439. Similar to cells with genetically inhibited Aurora-B, these ZM-treated G2/M-arrested cells accumulate 4N/8N DNA and produce multipolar spindles. The treatment with ZM-447439 causes cell death. The reduction in Akt phosphorylation at Ser473 and its substrates GSK3α/β phosphorylation at Ser21 and Ser9 is potently correlated with ZM-447439 suppression of Aurora kinase[2].
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| ln Vivo |
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| Enzyme Assay |
In vitro kinase assays[1]
Recombinant Aurora A and B were expressed as NH2-terminal His6-tagged fusion proteins using a baculovirus expression system according to the manufacturer's instructions. Aurora A was purified by affinity chromatography using Ni-NTA agarose, and Aurora B was purified by ion exchange chromatography using CM Sepharose Fast Flow. 1 ng purified recombinant enzyme was 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 (Biotinyl-Ahx-tetra (LRRWSLG)), 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 was then incubated at RT for 60 min. Reactions were stopped by addition of 20% phosphoric acid, and the products were captured on P30 nitrocellulose filters and assayed for incorporation of 33P with a Betaplate™ counter. No enzyme and no compound control values were used to determine the concentration of ZM447439, which gave 50% inhibition of enzyme activity. |
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| Cell Assay |
Cell culture [1]
ZM447439 was dissolved in DMSO at 10 mM and stored at −20°C for up to 9 mo in individual aliquots to avoid freeze-thaw cycles, and was then freshly diluted in media. The IC50 values for Aurora A and B (∼100 nM) were determined at the Km for ATP (see above). However, because the cellular ATP concentration is ∼200-fold higher, and because ZM447439 is an ATP competitor, we used ZM447439 at a concentration of 2 μM in all the cell assays unless stated otherwise. DMSO was added to drug-free cultures to account for the solvent. Cell cycle analysis and cloning assays[1] DNA content and mitotic index measurements and synchronization of TA-HeLa cells at G1/S using a double thymidine block were done as described previously (Taylor and McKeon, 1997). To determine cloning efficiency, MCF7 cells were plated in phenol red free DME plus 5% stripped serum, and were then treated with or without the anti-estrogen ICI 182780 at 1 μM for 48 h. ZM447439 was then added at the indicated concentrations for 72 h. The cells were harvested, washed, and ∼400 cells plated in each well of a 6-well plate in complete media without ZM447439. After 10 d, the colonies were fixed, stained with crystal violet, and counted. The cloning efficiency represents the number of colonies on ZM447439-treated plates compared with DMSO-treated controls. |
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| Animal Protocol |
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| References | |||
| Additional Infomation |
ZM447439 is a member of the class of quinazolines that is quinazoline which is substituted at positions 4, 6 and 7 by a (4-benzamidophenyl)nitrilo group, methoxy group and a 3-(morpholin-4-yl)propoxy group, respectively. It is an ATP-competitive inhibitor of Aurora A and Aurora B kinases with IC50 of 110 nM and 130 nM, respectively. It has a role as an Aurora kinase inhibitor, an antineoplastic agent and an apoptosis inducer. It is a member of benzamides, a member of quinazolines, an aromatic ether, a member of morpholines, a polyether, a secondary amino compound and a tertiary amino compound.
Mitotic Aurora kinases are essential for accurate chromosome segregation during cell division. Forced overexpression of Aurora kinase results in centrosome amplification and multipolar spindles, causing aneuploidy, a hallmark of cancer. ZM447439 (ZM), an Aurora selective ATP-competitive inhibitor, interferes with the spindle integrity checkpoint and chromosome segregation. Here, we showed that inhibition of Aurora kinase by ZM reduced histone H3 phosphorylation at Ser10 in Hep2 carcinoma cells. Multipolar spindles were induced in these ZM-treated G(2)/M-arrested cells with accumulation of 4N/8N DNA, similar to cells with genetically suppressed Aurora-B. Cells subsequently underwent apoptosis, as assessed by cleavage of critical apoptotic associated protein PARP. Hep2 cells formed a tumor-like cell mass in 3-dimensional matrix culture; inhibition of Aurora kinase by ZM either destructed the preformed cell mass or prevented its formation, by inducing apoptotic cell death as stained for cleaved caspase-3. Lastly, ZM inhibition of Aurora kinase was potently in association with decrease of Akt phosphorylation at Ser473 and its substrates GSK3alpha/beta phosphorylation at Ser21 and Ser9. Together, we demonstrated that Aurora kinase served as a potential molecular target of ZM for more selective therapeutic cancer treatment.[2] |
| Molecular Formula |
C29H31N5O4
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| Molecular Weight |
513.59
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| Exact Mass |
513.237
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| Elemental Analysis |
C, 67.82; H, 6.08; N, 13.64; O, 12.46
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| CAS # |
331771-20-1
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| Related CAS # |
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| PubChem CID |
9914412
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| Appearance |
White to gray solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
639.7±55.0 °C at 760 mmHg
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| Melting Point |
117-120ºC
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| Flash Point |
340.7±31.5 °C
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| Vapour Pressure |
0.0±1.9 mmHg at 25°C
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| Index of Refraction |
1.664
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| LogP |
2.66
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
38
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| Complexity |
709
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
OGNYUTNQZVRGMN-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C29H31N5O4/c1-36-26-18-24-25(19-27(26)38-15-5-12-34-13-16-37-17-14-34)30-20-31-28(24)32-22-8-10-23(11-9-22)33-29(35)21-6-3-2-4-7-21/h2-4,6-11,18-20H,5,12-17H2,1H3,(H,33,35)(H,30,31,32)
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| Chemical Name |
N-(4-((6-methoxy-7-(3-morpholinopropoxy)quinazolin-4-yl)amino)phenyl)benzamide
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| Synonyms |
ZM-447439; ZM 447439; N-[4-[[6-METHOXY-7-[3-(4-MORPHOLINYL)PROPOXY]-4-QUINAZOLINYL]AMINO]PHENYL]BENZAMIDE; N-(4-((6-methoxy-7-(3-morpholinopropoxy)quinazolin-4-yl)amino)phenyl)benzamide; TCMDC-125873; C29H31N5O4; ZM447439.
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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 |
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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)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.87 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 25.0 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: ≥ 2.5 mg/mL (4.87 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. View More
Solubility in Formulation 3: 30% PEG400+0.5% Tween80+5% propylene glycol:30mg/mL |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9471 mL | 9.7354 mL | 19.4708 mL | |
| 5 mM | 0.3894 mL | 1.9471 mL | 3.8942 mL | |
| 10 mM | 0.1947 mL | 0.9735 mL | 1.9471 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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