| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
BAY-320 targets the human Bub1 kinase, a serine/threonine protein kinase that plays a critical role in the spindle assembly checkpoint (SAC) during mitosis. Bub1 is essential for proper chromosome segregation and genomic stability. By inhibiting Bub1, BAY-320 disrupts the mitotic checkpoint, leading to chromosome missegregation and ultimately cell death in rapidly dividing cancer cells. The compound has an IC50 of 680 nM for human Bub1 in the presence of 2 mM ATP.
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| ln Vitro |
To address the role of Bub1 kinase activity in mammalian mitosis, we have made use of two novel small molecule inhibitors, BAY-320 and BAY-524. Using biochemical and cellular assays, we show that these ATP-competitive inhibitors potently and specifically block human Bub1 both in vitro and in living cells. By comparing phenotypes provoked by Bub1 kinase inhibition and Bub1 protein depletion, we are able to differentiate between catalytic and non-catalytic functions of Bub1. Our data indicate that Bub1 catalytic activity is largely dispensable for chromosome alignment and SAC function, arguing that Bub1 largely operates as a scaffolding protein. However, even though Bub1 inhibition per se exerts only minor effects on mitotic fidelity, BAY-320 and BAY-524 treatment sensitizes cells to clinically relevant low doses of Paclitaxel, resulting in remarkable impairment of chromosome segregation and cell proliferation.
In vitro, BAY-320 demonstrates potent inhibition of human Bub1 kinase activity with an IC50 of 680 nM in the presence of 2 mM ATP. The compound's activity has been characterized using biochemical kinase assays. As a Bub1 inhibitor, BAY-320 is used in research to study the function of Bub1 in the mitotic checkpoint and to evaluate the potential of Bub1 inhibition as an anticancer strategy. Specific cellular activity data, such as IC50 values for inhibition of cell proliferation in cancer cell lines, are not detailed in the available sources. |
| ln Vivo |
Specific in vivo activity data for BAY-320 are not detailed in the available sources. The compound's primary characterization comes from in vitro biochemical kinase assays. As a Bub1 inhibitor with potential anticancer activity, it may be evaluated in xenograft models of cancer to assess its in vivo efficacy. However, specific animal model studies have not been described in the available literature. The compound is intended for research use only.
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| Enzyme Assay |
The Bub1 kinase inhibition assay for BAY-320 involves incubating the compound with recombinant human Bub1 kinase, ATP, and a peptide substrate. The reaction is carried out in a kinase buffer optimized for Bub1 activity at a specified ATP concentration (e.g., 2 mM). After incubation at room temperature for a suitable period, the reaction is stopped, and the amount of phosphorylated substrate is quantified using a luminescence-based kinase assay (such as ADP-Glo) or a radiometric assay. IC50 values are calculated from dose-response curves generated with varying concentrations of the compound.
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| Cell Assay |
IC50 isobolograms
BAY-320 plus Paclitaxel combination studies were conducted with HeLa and NCI-H1299 cells. Cells were plated into 384-well plates at 600 (HeLa) or 700 (NCI-H1299) cells per well. After 24 hr, cells were treated with BAY-320 (concentration range, 1E-07 M to 1E-05 M) and Paclitaxel (concentration range, 1E-09 M to 1E-07 M) for single compound treatments, and in nine different fixed-ratio combinations of BAY-320 (D1) and Paclitaxel (D2) (0.9xD1+0.1xD2, 0.8xD1+0.2xD2, 0.7xD1+0.3xD2, 0.6xD1+0.4xD2, 0.5xD1+0.5xD2, 0.4xD1+0.6xD2, 0.3xD1+0.7xD2, 0.2xD1+0.8xD2, 0.1xD1+0.9xD2). Cell viability was assessed after 96 hr exposure, using the Cell Titre-Glo Luminescent Cell Viability Assay (Promega).
Specific cellular assay protocols for BAY-320 are not detailed in the available sources. To evaluate the cellular effects of Bub1 inhibition, cancer cells can be treated with BAY-320 at various concentrations, and cell proliferation can be measured using an MTT or SRB assay after 72 hours of treatment. The compound's effect on mitotic progression can be assessed by analyzing cell cycle distribution using flow cytometry. Chromosome missegregation can be evaluated by monitoring micronuclei formation or analyzing chromosome alignment in mitotic cells using immunofluorescence microscopy. |
| Animal Protocol |
Specific in vivo animal experiment protocols for BAY-320 are not detailed in the available sources. As a Bub1 inhibitor with potential anticancer activity, the compound would typically be evaluated in xenograft models using human cancer cell lines implanted subcutaneously into immunodeficient mice. BAY-320 would be administered orally or intraperitoneally, and tumor growth inhibition would be measured. However, such studies have not been described in the available literature.
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| ADME/Pharmacokinetics |
Specific pharmacokinetic data for BAY-320 are not provided in the available sources. The compound is a small molecule with a molecular weight of 492.52 and is soluble in DMSO at 83.33 mg/mL. It is stable as a powder at -20°C for 3 years and at 4°C for 2 years. In solvent, it can be stored at -80°C for 6 months or at -20°C for 1 month. Standard pharmacokinetic studies would typically involve administering the compound to rodents and measuring plasma concentrations over time using LC-MS/MS.
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| Toxicity/Toxicokinetics |
Specific toxicity data for BAY-320 are not provided in the available sources. As a research compound, it is intended for laboratory use only and is not approved for human therapeutic applications. The compound is available with a purity of ≥98%. Standard safety precautions should be followed when handling this compound, including the use of appropriate personal protective equipment.
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| References | |
| Additional Infomation |
BAY-320 is a Bub1 inhibitor with an IC50 of 680 nM for human Bub1 in the presence of 2 mM ATP. It shows potential as a targeted therapy for the treatment of various types of cancer where dysregulated cell division is a hallmark feature. The compound has a molecular formula of C26H26F2N6O2 and a molecular weight of 492.52. It is supplied for research purposes with a purity of ≥98% and is typically stored as a powder at -20°C.
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| Molecular Formula |
C26H26F2N6O2
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| Molecular Weight |
492.520452022552
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| Exact Mass |
492.208
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| CAS # |
1445830-50-1
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| PubChem CID |
71611462
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| Appearance |
White to off-white solid powder
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| LogP |
4.1
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
36
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| Complexity |
687
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCOC1=CC(=C(C(=C1)F)CN2C(=C(C(=N2)C3=NC=C(C(=N3)NC4=CC=NC=C4)OC)C)C5CC5)F
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| InChi Key |
WAELFQHBZPHEMW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H26F2N6O2/c1-4-36-18-11-20(27)19(21(28)12-18)14-34-24(16-5-6-16)15(2)23(33-34)26-30-13-22(35-3)25(32-26)31-17-7-9-29-10-8-17/h7-13,16H,4-6,14H2,1-3H3,(H,29,30,31,32)
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| Chemical Name |
2-[5-cyclopropyl-1-[(4-ethoxy-2,6-difluorophenyl)methyl]-4-methylpyrazol-3-yl]-5-methoxy-N-pyridin-4-ylpyrimidin-4-amine
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| Synonyms |
BAY-320; BAY 320; BAY320
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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 |
| 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 : ~83.33 mg/mL (~169.19 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.22 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 (4.22 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 | 2.0304 mL | 10.1519 mL | 20.3037 mL | |
| 5 mM | 0.4061 mL | 2.0304 mL | 4.0607 mL | |
| 10 mM | 0.2030 mL | 1.0152 mL | 2.0304 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.
BAY-320 and BAY-524 inhibit Bub1 kinase.Elife. 2016 Feb 17;5. pii: e12187. |
Inhibition of Bub1 affects Sgo1 and Sgo2 localization and chromatid cohesion.Elife. 2016 Feb 17;5. pii: e12187. |
Bub1 inhibition affects localization and activity of the CPC.Elife. 2016 Feb 17;5. pii: e12187. |
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