| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Nek2 (Never in mitosis gene A-related kinase 2). Nek2 is a serine/threonine kinase that plays a critical role in centrosome duplication and spindle assembly during mitosis. Overexpression of Nek2 has been observed in various human cancers, making it an attractive target for anticancer drug development. rac-CCT-250863 inhibits Nek2 activity by competing with ATP binding at the kinase domain.
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| ln Vitro |
In vitro, rac-CCT-250863 demonstrates potent inhibition of Nek2 kinase activity with an IC50 of 73 nM in enzymatic assays. The compound shows selectivity for Nek2 over other kinases including PLK1, MPS1, Cdk2, and Aurora A. In cell-based assays, rac-CCT-250863 induces cell cycle arrest and exhibits anti-proliferative activity in cancer cell lines. When combined with pomalidomide, the compound induces apoptosis.
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| ln Vivo |
In vivo, rac-CCT-250863 has been evaluated in preclinical models of cancer. The compound demonstrates anti-tumor activity by inducing cell cycle arrest and apoptosis in tumor xenografts. Its ability to inhibit Nek2 leads to mitotic defects and subsequent cell death in rapidly dividing cancer cells. The compound's efficacy is being investigated in various cancer models to establish its potential as a therapeutic agent for Nek2-overexpressing tumors.
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| Enzyme Assay |
Cell-free kinase assays for rac-CCT-250863 typically use recombinant human Nek2 protein and a peptide substrate in the presence of ATP. The compound is incubated with the enzyme at varying concentrations (0.1-10000 nM) for 30-60 minutes at room temperature. Kinase activity is measured using a luminescent ADP detection assay or radioactive ³³P-ATP incorporation. IC50 values are calculated from dose-response curves. Selectivity profiling is performed against a panel of kinases (e.g., PLK1, MPS1, Cdk2, Aurora A) using the same assay format to assess specificity.
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| Cell Assay |
Cellular assays are performed using cancer cell lines (e.g., HeLa, MCF-7, or other Nek2-overexpressing cells) cultured in appropriate media with 10% FBS. Cells are seeded in 96-well plates and treated with rac-CCT-250863 at concentrations ranging from 0.01-100 μM for 24-72 hours. Cell viability is assessed using MTT, CCK-8, or CellTiter-Glo assays to determine IC50 values for anti-proliferative activity. Cell cycle analysis is performed by flow cytometry after propidium iodide staining. Apoptosis is quantified using Annexin V/PI double staining or caspase-3/7 activity assays. Combination studies with other agents such as pomalidomide can be performed to assess synergistic effects.
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| Animal Protocol |
In vivo efficacy studies are conducted in immunodeficient mice bearing subcutaneous tumor xenografts derived from cancer cell lines. rac-CCT-250863 is administered via oral gavage or intraperitoneal injection at doses typically ranging from 10-100 mg/kg, daily or on a scheduled regimen. Tumor volume is measured every 2-3 days using calipers. At study termination, tumors are excised, weighed, and processed for immunohistochemistry or Western blot analysis to assess Nek2 inhibition and downstream markers of cell cycle arrest and apoptosis. Pharmacodynamic studies involve collecting tumors at various time points post-dose to measure target engagement and pathway modulation.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of rac-CCT-250863 are consistent with small molecule kinase inhibitors. The compound is expected to have moderate oral bioavailability and reasonable plasma half-life for in vivo efficacy studies. As an ATP-competitive kinase inhibitor, its distribution is likely to include significant tissue penetration. Detailed PK parameters (Cmax, Tmax, AUC, t1/2, clearance, volume of distribution) should be determined experimentally for each route of administration in the relevant preclinical species. Metabolic stability is assessed using liver microsome or hepatocyte incubation studies.
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| Toxicity/Toxicokinetics |
Toxicological data for rac-CCT-250863 are limited to preclinical research studies. As a Nek2 inhibitor, its toxicity profile is expected to be related to its mechanism of action, with effects on rapidly dividing tissues such as bone marrow, gastrointestinal epithelium, and hair follicles. Standard toxicology studies (acute, subchronic, and chronic) are required for clinical development. At research-grade doses used for in vitro and in vivo studies, the compound is handled with standard laboratory safety precautions.
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| Additional Infomation |
rac-CCT-250863 is a research tool for studying Nek2 biology and validating Nek2 as a therapeutic target in cancer. It is not an approved drug and has not entered clinical trials. The compound is available from multiple chemical suppliers for research purposes only. Its selectivity profile and potent activity make it a valuable chemical probe for dissecting Nek2-dependent cellular processes. The compound should be stored at -20°C and protected from light for long-term stability.
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| Molecular Formula |
C24H25F3N4O2S
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|---|---|
| Molecular Weight |
490.54111456871
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| Exact Mass |
490.165
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| CAS # |
1364269-06-6
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| PubChem CID |
57386035
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| Appearance |
White to off-white solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
582.9±50.0 °C at 760 mmHg
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| Flash Point |
306.3±30.1 °C
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| Vapour Pressure |
0.0±1.6 mmHg at 25°C
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| Index of Refraction |
1.591
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| LogP |
4.42
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
34
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| Complexity |
708
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(N)(=O)C1=CC=C(C2=CC(C3SC=C(CN(C)C)C=3)=CN=C2N)C=C1OC(C)/C=C\C(F)(F)F
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| InChi Key |
LLEOOSLUHRQCHZ-SREVYHEPSA-N
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| InChi Code |
InChI=1S/C24H25F3N4O2S/c1-14(6-7-24(25,26)27)33-20-10-16(4-5-18(20)23(29)32)19-9-17(11-30-22(19)28)21-8-15(13-34-21)12-31(2)3/h4-11,13-14H,12H2,1-3H3,(H2,28,30)(H2,29,32)/b7-6-
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| Chemical Name |
4-[2-amino-5-[4-[(dimethylamino)methyl]thiophen-2-yl]pyridin-3-yl]-2-[(Z)-5,5,5-trifluoropent-3-en-2-yl]oxybenzamide
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 2.0386 mL | 10.1928 mL | 20.3857 mL | |
| 5 mM | 0.4077 mL | 2.0386 mL | 4.0771 mL | |
| 10 mM | 0.2039 mL | 1.0193 mL | 2.0386 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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