Ro-3306

Catalog No.S7747

For research use only.

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.

Ro-3306 Chemical Structure

CAS No. 872573-93-8

Selleck's Ro-3306 has been cited by 93 publications

Purity & Quality Control

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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 MUTxTHRUKGG|c3H5 MWrQMYdtgWOxcILveIVqdiC|dXLzeJJifGW|IHnk[Y51cW[rZXSgbY4hU0JvMz2xJIFl\W6xY3HyZ4lvd22jIHPlcIwhdGmwZTygdWhVWyC2aHXyZZBmfXSrYzDsbYJz[XK7IIPjdoVmdg>? MXO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8{OTVzNUK4OEc,OzF3MUWyPFQ9N2F-
human H460a cells MnvlVJJwdGmoZYLheIlwdiCjc4PhfS=> M3r0XWFvfGmycn;sbYZmemG2aY\lJIFkfGm4aYT5JIFo[Wmwc4SgbJVu[W5iSES2NIEh[2WubIOsJGlEPTB;MT6wOEDPxE1? M13CUFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF5M{CzOFIyLz5zN{OwN|QzOTxxYU6=
human MDA435 cells MXLQdo9tcW[ncnH0bY9vKGG|c3H5 MmDFRY51cXC{b3zp[oVz[XSrdnWgZYN1cX[rdImgZYdicW6|dDDoeY1idiCPRFG0N|Uh[2WubIOsJGlEPTB;MT6xOEDPxE1? MnHyQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd|MEO0NlEoRjF5M{CzOFIyRC:jPh?=
human HCT116 cells M4XDVXBzd2yrZnXyZZRqd25iYYPzZZk> MoD4RY51cXC{b3zp[oVz[XSrdnWgZYN1cX[rdImgZYdicW6|dDDoeY1idiCKQ2SxNVYh[2WubIOsJGlEPTB;MT6xOEDPxE1? MVO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPzNyM{SyNUc,OTd|MEO0NlE9N2F-
human RKO cells NX\UbpVGWHKxbHnm[ZJifGmxbjDhd5NigQ>? MWjBcpRqeHKxbHnm[ZJifGm4ZTDhZ5Rqfmm2eTDh[4FqdnO2IHj1cYFvKFKNTzDj[YxteyxiSVO1NF0zKM7:TR?= NV\HVHdNRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUezNFM1OjFpPkG3N|A{PDJzPD;hQi=>
human SW480 cells NHTmXGhRem:uaX\ldoF1cW:wIHHzd4F6 NUnSNY5YSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDTW|Q5OCClZXzsd{whUUN3ME2yMlIh|ryP MXy8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPzNyM{SyNUc,OTd|MEO0NlE9N2F-
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human HCT116 cells NF\oe5hEgXSxdH;4bYPDqGG|c3H5 MWW1JIRigXN? M{XlbmN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJGhEXDFzNjDj[YxteyCjZoTldkA2KGSjeYOgZpkhVVSVIHHzd4F697zOIFnDOVA:Oy5{IN88US=> M1;6OlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{M{K2NVY5Lz5{MkOyOlE3QDxxYU6=
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KYSE-520 M13vNWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M{fGVmlvcGmkaYTpc44hd2ZiaIXtZY4hU1mVRT21NlAh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NEA:KDR4LkW3NlEh|ryPLh?= M{KzdVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFM6Ozl{OT:nQnNCVkeHUkyvZV4>
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EPLC-272H NEjXbFVIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NGnDTYJKdmirYnn0bY9vKG:oIHj1cYFvKEWSTFOtNlczUCClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LDDJR|UxKD1iNE[uO|I5OyEQvF2u M1zxOFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFM6Ozl{OT:nQnNCVkeHUkyvZV4>
T-24 NF\LXnVIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NWfTbpZJUW6qaXLpeIlwdiCxZjDoeY1idiCWLUK0JINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUCgQUA1Py57MkmyJO69VS5? M2jlU|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFM6Ozl{OT:nQnNCVkeHUkyvZV4>
BHY NIjTfnZIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NFfZfldKdmirYnn0bY9vKG:oIHj1cYFvKEKKWTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyIE2gOFcvQTl7IN88UU4> MkDBQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMN|k{QTJ7Lze+V2FPT0WUPD;hQi=>
NCI-H82 M4fqSmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NYPqZWtFUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFgzKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XluIFnDOVAhRSB2OD6yNVch|ryPLh?= NHnxNWU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmw{QTN7MkmvK|5USU6JRWK8M4E,
SF268 MlrZS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? M1XHRmlvcGmkaYTpc44hd2ZiaIXtZY4hW0Z{NkigZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3MDC9JFQ5NjZzOEmg{txONg>? MXu8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyzPVM6OjlxJ{7TRW5ITVJ:L3G+
C-33-A MnzUS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NV\wfYR1UW6qaXLpeIlwdiCxZjDoeY1idiCFLUOzMWEh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NEA:KDR6Lke1OVQh|ryPLh?= NWLJXY5MRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOzl|OUK5M{c,W0GQR1XSQE9iRg>?
MIA-PaCa-2 MkToS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NVv0SFhlUW6qaXLpeIlwdiCxZjDoeY1idiCPSVGtVIFE[S1{IHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO|YYmsJGlEPTBiPTC0PE45Ozh6IN88UU4> NYDlVJZnRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOzl|OUK5M{c,W0GQR1XSQE9iRg>?
HCC1954 M4jwNmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M3HIU2lvcGmkaYTpc44hd2ZiaIXtZY4hUEOFMUm1OEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwJF0hPDlwMEi3NUDPxE1w MYO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyzPVM6OjlxJ{7TRW5ITVJ:L3G+
NCI-H1666 MlzoS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? Ml7lTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG2OlYh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NEA:KDR7LkO2OFIh|ryPLh?= MVm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyzPVM6OjlxJ{7TRW5ITVJ:L3G+
GR-ST MXnHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MnrsTY5pcWKrdHnvckBw\iCqdX3hckBIWi2VVDDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyIE2gOFkvOzZ7NTFOwG0v M3P0bFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFM6Ozl{OT:nQnNCVkeHUkyvZV4>
Ca9-22 MlHvS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NYLUWHpVUW6qaXLpeIlwdiCxZjDoeY1idiCFYUmtNlIh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NEA:KDR7LkSwNVMh|ryPLh?= MWK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyzPVM6OjlxJ{7TRW5ITVJ:L3G+
D-502MG M3rGR2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NIH4[3pKdmirYnn0bY9vKG:oIHj1cYFvKERvNUCyUWch[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NEA:KDR7LkSxO{DPxE1w MlHNQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMN|k{QTJ7Lze+V2FPT0WUPD;hQi=>
SCC-4 M{m1O2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MmPmTY5pcWKrdHnvckBw\iCqdX3hckBUS0NvNDDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyIE2gOFkvPTZyMjFOwG0v Mmr6QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMN|k{QTJ7Lze+V2FPT0WUPD;hQi=>
RKO M2L3b2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NEnJV5FKdmirYnn0bY9vKG:oIHj1cYFvKFKNTzDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyIE2gOFkvQDN3NjFOwG0v M1rmblxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFM6Ozl{OT:nQnNCVkeHUkyvZV4>
Assay
Methods Test Index PMID
Western blot p-Cdk1 / Cdk1 / Cleaved PARP / Cleaved caspase-3 / p-p38 / Bad / Apaf1-1 / p-H2Ax / Bim / Bad 27385216
Immunofluorescence PARP1 / Plk1 28069876
Growth inhibition assay Cell viability 27385216

Protocol (from reference)

Kinase Assay:[1]
  • 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]
  • 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.

Solubility (25°C)

In vitro

Chemical Information

Molecular Weight 351.45
Formula

C18H13N3OS2

CAS No. 872573-93-8
Storage 3 years -20°C powder
2 years -80°C in solvent
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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% DMSO % % Tween 80 % ddH2O
%DMSO %

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.

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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