Ro 3306 |
| Katalog-Nr.GC12348 |
Ro 3306 ist ein potenter und selektiver Inhibitor von CDK1 mit Kis von 20 nM, 35 nM und 340 nM für CDK1, CDK1/Cyclin B1 bzw. CDK2/Cyclin E.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 872573-93-8
Sample solution is provided at 25 µL, 10mM.
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RO-3306 is an ATP-competitive, potent cyclin-dependent kinase (CDK) 1 inhibitor with Ki values of 35 and 110 nM for cdk1/cyclin B1 and cdk1/cyclin A, respectively.
Cell cycle progression is regulated by a series of CDKs. Cdk1 controls the cell cycle entry into mitosis. It forms a bipartite complex with cyclin B and phosphorylates a spectrum of substrates that coordinate nuclear envelope breakdown, chromosome condensation, assembly of the mitotic spindle and activation of the spindle assembly checkpoint.
CDK1 inhibitor RO-3306 was reported to impair BRCA1 localization to DSBs, and consistent with this, suppressed RAD51 focus formation in DU145 cells when applied at the GI50 (1μM). Co-treatment of 100 nM AZ12253801 and 1 μM RO-3306 manifested a marked increase in pre-G1
DNA, indicating apoptosis induction. In viability assays, RO-3306 induced a significant shift to the left of the AZ12253801 dose-response curve, with 2.4-fold reduction in GI50, comparable to RAD51 depletion or inhibition. At sub-50nM AZ12253801 concentrations there was more striking sensitization, with 13-fold reduction in GI80. Paralleling effects of RAD51 knockdown, RO-3306 did not affect AZ12253801 sensitivity of PC3 or LNCaP cells, but did sensitize 22Rv1 cells.
References:
[1]. Lodhia KA, Gao S, Aleksic T, et al. Suppression of homologous recombination sensitizes human tumor cells to IGF-1R inhibition. Int J Cancer. 2014 Nov 12.
| Kinase experiment [1]: | |
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Binding assays |
The CDK assays were run by using recombinant human CDK/cyclin complexes (CDK1/cyclin B1, CDK1/cyclin A, CDK2/cyclin E, and CDK4/cyclin D) expressed and isolated from Hi5 insect cells. The activity of CDK1/cyclin B1, CDK1/cyclin A, CDK2/cyclin E, and CDK4/cyclin D was measured by a homogeneous time-resolved fluorescence assay in a 96-well format. The assay buffer contained 25 mM Hepes, 6.25 mM MgCl2, 0.003% Tween 20, 0.3 mg/ml BSA, 1.5 mM DTT, and ATP as follows: 162 μM (CDK1), 90 μM (CDK2), or 135 μM (CDK4). CDK1 and CDK2 buffer contained 10 mM MgCl2. Test compounds were diluted in assay buffer to 3-fold their final concentration in 20 μl, and the reaction was started by the addition of a 40 μl assay buffer containing the pRB substrate (0.185 μM). The plates were incubated at 37°C for 30 min with constant agitation, and the reaction was terminated by the addition of 15 μl of 1.6 μM anti-phospho pRB antibody in 25 mM Hepes/24 mM EDTA/0.2 mg/ml BSA. After an additional 30 min of incubation with shaking, 15 μl of 3 nM Lance-Eu-W1024-labeled anti-rabbit IgG and 60 nM Alophycocyanin-conjugated anti-His-6 antibody in 25 mM Hepes/0.5 mg/ml BSA was added and incubated for 1 h. The plates were read in the Victor-V multilabel reader at excitation 340 nm and emission 615 nm and 665 nm. The IC50 values were calculated from the readings at 665 nm and normalized for Europium readings at 615 nm. |
| Cell experiment [1]: | |
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Cell lines |
Proliferating human cells HCT116, SW480, Hela |
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Preparation method |
The solubility of this compound in DMSO is >4.4 mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
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Reacting condition |
9 μM, 20 h, |
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Applications |
Treatment of proliferating human cancer cells (HCT116, SW480, and HeLa) with RO-3306 for 20 h completely blocked the cell cycle in the G2/M phase. In cancer cell lines (RKO, SJSA, MDAMB-435, and DU145) RO-3306 provided an effective means for cell synchronization in the late G2 phase. HeLa cells were enriched in mitotic cells by release from RO-3306 block (9 μM for 18 h) and followed for morphological changes in the absence or presence of 9 μM RO-3306. Treatment with 4 μM RO-3306 for 48 h induced apoptosis in cancer cells. |
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Other notes |
Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal. |
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References: [1]. Vassilev L T, Tovar C, Chen S, et al. Selective small-molecule inhibitor reveals critical mitotic functions of human CDK1[J]. Proceedings of the National Academy of Sciences, 2006, 103(28): 10660-10665. |
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| Cas No. | 872573-93-8 | SDF | |
| Chemical Name | (Z)-5-(quinolin-6-ylmethylene)-2-((thiophen-2-ylmethyl)amino)thiazol-4(5H)-one | ||
| Canonical SMILES | O=C1N=C(NCC2=CC=CS2)S/C1=C\C3=CC=C(N=CC=C4)C4=C3 | ||
| Formula | C18H13N3OS2 | M.Wt | 351.45 |
| Löslichkeit | ≥ 4.39mg/mL in DMSO | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.8454 mL | 14.2268 mL | 28.4535 mL |
| 5 mM | 0.5691 mL | 2.8454 mL | 5.6907 mL |
| 10 mM | 0.2845 mL | 1.4227 mL | 2.8454 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 33 reference(s) in Google Scholar.)
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