Ro-3306

For research use only.

Catalog No.S7747

80 publications

Ro-3306 Chemical Structure

CAS No. 872573-93-8

RO-3306 is an ATP-competitive, and selective CDK1 inhibitor with Ki of 20 nM, >15-fold selectivity against a diverse panel of human kinases. RO-3306 enhances p53-mediated Bax activation and mitochondrial apoptosis.

Selleck's Ro-3306 has been cited by 80 publications

Purity & Quality Control

Choose Selective CDK Inhibitors

Biological Activity

Description RO-3306 is an ATP-competitive, and selective CDK1 inhibitor with Ki of 20 nM, >15-fold selectivity against a diverse panel of human kinases. RO-3306 enhances p53-mediated Bax activation and mitochondrial apoptosis.
Targets
CDK1 [1]
(Cell-free assay)
PKCδ [1]
(Cell-free assay)
SGK [1]
(Cell-free assay)
ERK [1]
(Cell-free assay)
20 nM(Ki) 318 nM(Ki) 497 nM(Ki) 1980 nM(Ki)
In vitro

RO-3306 inhibits CDK1/cyclin B1, CDK1/cyclin A, CDK2/cyclin E, and CDK4/cyclin D activity with Ki of 35 nM, 110 nM, 340 nM, and over 2000 nM, respectively. Treatment of HCT116, SW480, and HeLa cells with RO-3306 for 20 h leads to a complete block of the cell cycle in the G2/M phase. The proliferation of both HCT116 and SW480 is effectively blocked by RO-3306. RO-3306 appears to be more proapoptotic in cancer cells (HCT116 and SW480) than nontumorigenic cells (MCF 10A and MCF 12A).[1] RO-3306 effectively arrests oocyte maturation at a concentration of 10 μM.[2]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
KB-3-1 adenocarcinoma cell line MVjxTHRUKGG|c3H5 NHPp[XRRNWeueXPvdJJwfGWrbjDzeYJ{fHKjdHXzJIll\W62aX\p[YQhcW5iS1KtN{0yKGGmZX7vZ4Fz[2mwb33hJINmdGxibHnu[UwheUiWUzD0bIVz[XCndYTpZ{BtcWK{YYL5JJNkemWnbh?= NXn0fWJwRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxM{G1NVUzQDRpPkOxOVE2Ojh2PD;hQi=>
human H460a cells NGLNZ4ZRem:uaX\ldoF1cW:wIHHzd4F6 MoH3RY51cXC{b3zp[oVz[XSrdnWgZYN1cX[rdImgZYdicW6|dDDoeY1idiCKNE[wZUBk\WyuczygTWM2OD1zLkC0JO69VQ>? NGTsO249[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zN{OwN|QzOSd-MUezNFM1OjF:L3G+
human MDA435 cells M{XqTnBzd2yrZnXyZZRqd25iYYPzZZk> NGm2fnNCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIF3ERVQ{PSClZXzsd{whUUN3ME2xMlE1KM7:TR?= MUm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPzNyM{SyNUc,OTd|MEO0NlE9N2F-
human HCT116 cells NHPyU3RRem:uaX\ldoF1cW:wIHHzd4F6 M{fZUWFvfGmycn;sbYZmemG2aY\lJIFkfGm4aYT5JIFo[Wmwc4SgbJVu[W5iSFPUNVE3KGOnbHzzMEBKSzVyPUGuNVQh|ryP NGTqNVM9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zN{OwN|QzOSd-MUezNFM1OjF:L3G+
human RKO cells MUPQdo9tcW[ncnH0bY9vKGG|c3H5 NV7xOJROSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDST28h[2WubIOsJGlEPTB;MjFOwG0> NUXufGtCRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUezNFM1OjFpPkG3N|A{PDJzPD;hQi=>
human SW480 cells NWCxWFhZWHKxbHnm[ZJifGmxbjDhd5NigQ>? NETYZoVCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIGPXOFgxKGOnbHzzMEBKSzVyPUKuNkDPxE1? MX28ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPzNyM{SyNUc,OTd|MEO0NlE9N2F-
human SJSA1 cells NYHjNHZ6WHKxbHnm[ZJifGmxbjDhd5NigQ>? Mkj0RY51cXC{b3zp[oVz[XSrdnWgZYN1cX[rdImgZYdicW6|dDDoeY1idiCVSmPBNUBk\WyuczygTWM2OD12LkW1JO69VQ>? Mn3tQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd|MEO0NlEoRjF5M{CzOFIyRC:jPh?=
human HCT116 cells NXnpXlVXS3m2b4TvfIlkyqCjc4PhfS=> M1XkW|Uh\GG7cx?= NX23bYQ4S3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hUEOWMUG2JINmdGy|IHHmeIVzKDViZHH5d{BjgSCPVGOgZZN{[XoxvJygTWM2OD1|LkKg{txO M{LpUVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{M{K2NVY5Lz5{MkOyOlE3QDxxYU6=
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C-33-A M2PWbGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MYrJcohq[mm2aX;uJI9nKGi3bXHuJGMuOzNvQTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyIE2gOFgvPzV3NDFOwG0v NXHXe3J1RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOzl|OUK5M{c,W0GQR1XSQE9iRg>?
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HCC1954 NVW4UlgzT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MWrJcohq[mm2aX;uJI9nKGi3bXHuJGhESzF7NUSgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3MDC9JFQ6NjB6N{Gg{txONg>? Ml63QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMN|k{QTJ7Lze+V2FPT0WUPD;hQi=>
NCI-H1666 NXvPZXB5T3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NWDPVolIUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFE3PjZiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeTygTWM2OCB;IES5MlM3PDJizszNMi=> M3[1OlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFM6Ozl{OT:nQnNCVkeHUkyvZV4>
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SCC-4 M4HDbWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NIXyfmtKdmirYnn0bY9vKG:oIHj1cYFvKFOFQz20JINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUCgQUA1QS53NkCyJO69VS5? NFLlTVk9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmw{QTN7MkmvK|5USU6JRWK8M4E,
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... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
p-Cdk1 / Cdk1 / Cleaved PARP / Cleaved caspase-3 / p-p38 / Bad / Apaf1-1 / p-H2Ax / Bim / Bad; 

PubMed: 27385216     


OVCA-429 and OVCAR-3 were exposed to RO-3306 for 24 h in indicated dose dependent manner, (F) 10 uM RO-3306 for 12 h up to 48 h. The expression level of the indicated protein and the phosphorylated form of H2A.x and p38 were examined via Western blot analyses.

27385216
Immunofluorescence
PARP1 / Plk1 ; 

PubMed: 28069876     


Representative images of IF staining for PARP1 and Plk1 in 22RV1 cells treated with 0.3 mM mimosine for 20 h, 4 mM hydroxyurea for 24 h, 10 µM RO-3306 for 7 h or 200 ng/ml nocodazole (Noc) for 12 h.

28069876
Growth inhibition assay
Cell viability; 

PubMed: 27385216     


OVCA-429 and OVCAR-3 were exposed to RO-3306 in indicated time- and dose-dependent manner. Cell viability were measured via crystal violet assay. Results are the means ± S.E. n = 3. 

27385216

Protocol

Kinase Assay:[1]
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CDK assay:

The activity of CDK1 cyclin B1, CDK1 cyclin A, CDK2 cyclin E, and CDK4 cyclin D is 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 mM (CDK2), or 135 μM (CDK4). CDK1 and CDK2 buffer contained 10 mM MgCl2. Test compounds are diluted in assay buffer to 3-fold their final concentration in 20 μL, and the reaction is started by the addition of a 40 μL assay buffer containing the pRB substrate (0.185μM). The plates are incubated at 37°C for 30 min with constant agitation, and the reaction is terminated by the addition of 15 μL of 1.6μM anti-phospho pRB antibody (Ser-780) in 25 mM Hepes, 24 mM EDTA, and 0.2 mg/mL BSA. After an additional 30 min of incubation with shaking, 15μL of 3nM Lance-Eu-W1024-labeledanti-rabbitIgG and 60 nM Alophycocyanin-conjugated anti-His-6 antibody in 25 mM Hepes, and 0.5 mg/mL BSA is added and incubated for 1 h. The plates are read in the Victor-V multi- label reader at excitation 340 nm and emission 615 nm and 665 nm. The IC50 values are calculated from the readings at 665 nm and normalized for Europium readings at 615 nm. Ki values are calculated according to the equation: Ki= IC50/(1 + S/Km ), where S is the ATP concentration in the assay and Km is the Michaelis-Menten constant for ATP. The inhibitory activity against the panel of kinases is determined by the IMAP assay technology.
Cell Research:[3]
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  • Cell lines: MDA-MB-231 cell line
  • Concentrations: 20 μM
  • Incubation Time: 72 h
  • Method: Log phases cells (25,000) are seeed in 96-well plates and incubated in a 37℃ incubator with CO2, After 24 h, different concentrations of RO-3306 are administered to determine the drug concentrations required to achieve a 50% growth inhibition (IC50). MTT (20 μL, 5mg/mL stock solution in saline) is added to each well and the cells are incubated for 4 h. Supernatants are removed and formazan crystals from viable cells are solubilized with 200 μL anhydrous DMSO. The absorbance is detected with a 550 model microplate reader at the 565 nm wavelength.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 13 mg/mL warmed (36.98 mM)
Water Insoluble
Ethanol 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.

Chemical Information

Molecular Weight 351.45
Formula

C18H13N3OS2

CAS No. 872573-93-8
Storage powder
in solvent
Synonyms N/A
Smiles C1=CC2=C(C=CC(=C2)C=C3C(=O)NC(=NCC4=CC=CS4)S3)N=C1

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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Step 2: Enter the in vivo formulation ()
% DMSO % % Tween 80 % ddH2O
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Bio Calculators

Molarity Calculator

Molarity Calculator

Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

Mass (mg) = Concentration (mM) × Volume (mL) × Molecular Weight (g/mol)

  • Mass
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*When preparing stock solutions, please always use the batch-specific molecular weight of the product found on the via label and SDS / COA (available on product pages).

Dilution Calculator

Dilution Calculator

Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

Concentration (start) x Volume (start) = Concentration (final) x Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2 ( Input Output )

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* When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and SDS / COA (available online).

The Serial Dilution Calculator Equation

  • Serial Dilutions

  • Computed Result

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
    C4=C3/X C4: LOG(C4):
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Molecular Weight Calculator

Molecular Weight Calculator

Enter the chemical formula of a compound to calculate its molar mass and elemental composition:

Total Molecular Weight: g/mol

Tip: Chemical formula is case sensitive. C10H16N2O2 c10h16n2o2

Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter its chemical formula and click 'Calculate'.

Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

Molarity Calculator

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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CDK Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID