Home Cell Cycle CDK inhibitor Roscovitine For research use only.

Roscovitine

Synonyms: CYC202, Seliciclib, R-roscovitine

Roscovitine is a potent and selective CDK inhibitor for Cdc2, CDK2 and CDK5 with IC50 of 0.65 μM, 0.7 μM and 0.16 μM in cell-free assays. It shows little effect on CDK4/6. Phase 2.

Roscovitine Chemical Structure

Roscovitine Chemical Structure

CAS No. 186692-46-6

Purity & Quality Control

Roscovitine Related Products

Signaling Pathway

CDK Signaling Pathways

CDK Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
LB771-HNC Growth Inhibition Assay IC50=48.9212 μM SANGER
LB2241-RCC Growth Inhibition Assay IC50=48.6202 μM SANGER
DU-4475 Growth Inhibition Assay IC50=48.4937 μM SANGER
LB2518-MEL Growth Inhibition Assay IC50=47.0448 μM SANGER
NCI-H209 Growth Inhibition Assay IC50=46.0115 μM SANGER
CGTH-W-1 Growth Inhibition Assay IC50=44.9697 μM SANGER
MS-1 Growth Inhibition Assay IC50=42.893 μM SANGER
GI-ME-N Growth Inhibition Assay IC50=42.6671 μM SANGER
DG-75 Growth Inhibition Assay IC50=42.6546 μM SANGER
MLMA Growth Inhibition Assay IC50=42.2787 μM SANGER
HT Growth Inhibition Assay IC50=42.0028 μM SANGER
LC-1F Growth Inhibition Assay IC50=41.5705 μM SANGER
NCI-H1882 Growth Inhibition Assay IC50=40.5998 μM SANGER
NTERA-S-cl-D1 Growth Inhibition Assay IC50=39.5842 μM SANGER
NCI-H345 Growth Inhibition Assay IC50=38.9106 μM SANGER
MONO-MAC-6 Growth Inhibition Assay IC50=38.2477 μM SANGER
RS4-11 Growth Inhibition Assay IC50=37.7069 μM SANGER
ML-2 Growth Inhibition Assay IC50=37.6712 μM SANGER
OPM-2 Growth Inhibition Assay IC50=37.2949 μM SANGER
LU-139 Growth Inhibition Assay IC50=37.1856 μM SANGER
COLO-684 Growth Inhibition Assay IC50=37.012 μM SANGER
MOLT-4 Growth Inhibition Assay IC50=36.3276 μM SANGER
TE-6 Growth Inhibition Assay IC50=36.3246 μM SANGER
TE-441-T Growth Inhibition Assay IC50=36.1148 μM SANGER
IMR-5 Growth Inhibition Assay IC50=35.3139 μM SANGER
K5 Growth Inhibition Assay IC50=35.0861 μM SANGER
TE-10 Growth Inhibition Assay IC50=34.9422 μM SANGER
NCI-H2141 Growth Inhibition Assay IC50=34.6533 μM SANGER
KGN Growth Inhibition Assay IC50=34.2524 μM SANGER
LP-1 Growth Inhibition Assay IC50=33.8908 μM SANGER
NCI-H64 Growth Inhibition Assay IC50=33.8597 μM SANGER
RKO Growth Inhibition Assay IC50=33.5969 μM SANGER
NCI-H526 Growth Inhibition Assay IC50=33.4936 μM SANGER
GOTO Growth Inhibition Assay IC50=32.9129 μM SANGER
Calu-6 Growth Inhibition Assay IC50=32.4745 μM SANGER
LOUCY Growth Inhibition Assay IC50=32.1253 μM SANGER
SK-N-FI Growth Inhibition Assay IC50=31.7535 μM SANGER
SIG-M5 Growth Inhibition Assay IC50=31.6833 μM SANGER
NKM-1 Growth Inhibition Assay IC50=31.1397 μM SANGER
NCI-SNU-1 Growth Inhibition Assay IC50=31.1059 μM SANGER
NCI-H82 Growth Inhibition Assay IC50=31.0135 μM SANGER
NCI-H510A Growth Inhibition Assay IC50=30.0329 μM SANGER
ES3 Growth Inhibition Assay IC50=29.9582 μM SANGER
BB30-HNC Growth Inhibition Assay IC50=29.9483 μM SANGER
KM12 Growth Inhibition Assay IC50=29.6239 μM SANGER
GI-1 Growth Inhibition Assay IC50=29.0113 μM SANGER
NOS-1 Growth Inhibition Assay IC50=28.9733 μM SANGER
TE-8 Growth Inhibition Assay IC50=28.908 μM SANGER
TE-9 Growth Inhibition Assay IC50=28.7969 μM SANGER
HL-60 Growth Inhibition Assay IC50=27.9869 μM SANGER
QIMR-WIL Growth Inhibition Assay IC50=27.9144 μM SANGER
KARPAS-299 Growth Inhibition Assay IC50=26.8646 μM SANGER
KURAMOCHI Growth Inhibition Assay IC50=26.8082 μM SANGER
BL-41 Growth Inhibition Assay IC50=25.9597 μM SANGER
NCI-H2126 Growth Inhibition Assay IC50=25.6529 μM SANGER
HOP-62 Growth Inhibition Assay IC50=25.4425 μM SANGER
IST-SL2 Growth Inhibition Assay IC50=24.5343 μM SANGER
HH Growth Inhibition Assay IC50=24.3819 μM SANGER
LS-513 Growth Inhibition Assay IC50=23.5179 μM SANGER
EB-3 Growth Inhibition Assay IC50=23.1831 μM SANGER
ACN Growth Inhibition Assay IC50=21.3389 μM SANGER
NOMO-1 Growth Inhibition Assay IC50=21.2008 μM SANGER
ES8 Growth Inhibition Assay IC50=21.06 μM SANGER
CESS Growth Inhibition Assay IC50=20.8549 μM SANGER
BL-70 Growth Inhibition Assay IC50=20.3274 μM SANGER
MHH-PREB-1 Growth Inhibition Assay IC50=20.0356 μM SANGER
BC-1 Growth Inhibition Assay IC50=19.1198 μM SANGER
LC4-1 Growth Inhibition Assay IC50=18.8734 μM SANGER
COLO-320-HSR Growth Inhibition Assay IC50=18.7688 μM SANGER
A101D Growth Inhibition Assay IC50=18.3208 μM SANGER
BC-3 Growth Inhibition Assay IC50=18.0305 μM SANGER
TGW Growth Inhibition Assay IC50=17.8124 μM SANGER
JAR Growth Inhibition Assay IC50=17.0152 μM SANGER
HD-MY-Z Growth Inhibition Assay IC50=16.8246 μM SANGER
NCI-H1304 Growth Inhibition Assay IC50=16.3601 μM SANGER
OS-RC-2 Growth Inhibition Assay IC50=15.8382 μM SANGER
OCI-AML2 Growth Inhibition Assay IC50=15.6482 μM SANGER
HCC1599 Growth Inhibition Assay IC50=14.5975 μM SANGER
SCC-3 Growth Inhibition Assay IC50=14.2956 μM SANGER
RPMI-6666 Growth Inhibition Assay IC50=13.9121 μM SANGER
MEG-01 Growth Inhibition Assay IC50=13.8379 μM SANGER
Raji Growth Inhibition Assay IC50=13.7894 μM SANGER
RPMI-8402 Growth Inhibition Assay IC50=13.6262 μM SANGER
GCIY Growth Inhibition Assay IC50=12.8613 μM SANGER
697 Growth Inhibition Assay IC50=12.6007 μM SANGER
D-247MG Growth Inhibition Assay IC50=12.3516 μM SANGER
NB1 Growth Inhibition Assay IC50=12.3308 μM SANGER
COR-L279 Growth Inhibition Assay IC50=12.2907 μM SANGER
LB831-BLC Growth Inhibition Assay IC50=11.5624 μM SANGER
ST486 Growth Inhibition Assay IC50=10.351 μM SANGER
SK-UT-1 Growth Inhibition Assay IC50=10.35 μM SANGER
BB65-RCC Growth Inhibition Assay IC50=9.97495 μM SANGER
KARPAS-422 Growth Inhibition Assay IC50=9.96336 μM SANGER
Becker Growth Inhibition Assay IC50=9.46082 μM SANGER
KS-1 Growth Inhibition Assay IC50=9.45785 μM SANGER
JiyoyeP-2003 Growth Inhibition Assay IC50=8.50264 μM SANGER
NCCIT Growth Inhibition Assay IC50=7.55482 μM SANGER
MRK-nu-1 Growth Inhibition Assay IC50=7.12969 μM SANGER
A3-KAW Growth Inhibition Assay IC50=5.76116 μM SANGER
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 15958589
LP-1 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human LP-1 cells at 30 uM after 3 hrs using TUNEL staining by flow cytometry 15958589
LP-1 Cytotoxicity assay 20 to 30 uM 24 hrs Cytotoxicity against human LP-1 cells assessed as reduction of cell viability at 20 to 30 uM treated for 24 hrs followed by washout measured after total 72 hrs growth period alamar blue assay relative to control 15958589
LP-1 Apoptosis assay 30 uM 1.5 hrs Induction of apoptosis in human LP-1 cells assessed as reduction of RNA polymerase 2 phosphoserine 2 level at 30 uM after 1.5 hrs by immunoblotting 15958589
LP-1 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human LP-1 cells assessed as reduction of Mcl-1 protein level at 30 uM after 3 hrs by immunoblotting 15958589
LP-1 Apoptosis assay 30 uM 3 to 5 hrs Induction of apoptosis in human LP-1 cells assessed as increase in level of cleaved PARP at 30 uM after 3 to 5 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 5 hrs Induction of apoptosis in human NCI-H929 cells assessed as increase in level of cleaved PARP at 30 uM after 5 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 1.5 hrs Induction of apoptosis in human NCI-H929 cells assessed as fast slow migrating hyperphosphorylated RNA polymerase 2O form at 30 uM after 1.5 hrs by immunoblotting 15958589
RPM18226 Apoptosis assay 30 uM 1.5 hrs Induction of apoptosis in human RPM18226 cells assessed as reduction of RNA polymerase 2 phosphoserine 2 level at 30 uM after 1.5 hrs by immunoblotting 15958589
RPM18226 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human RPM18226 cells assessed as reduction of Mcl-1 protein level at 30 uM after 3 hrs by immunoblotting 15958589
RPM18226 Apoptosis assay 30 uM 3 to 5 hrs Induction of apoptosis in human RPM18226 cells assessed as increase in level of cleaved PARP at 30 uM after 3 to 5 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human NCI-H929 cells assessed as changes in XIAP protein level at 30 uM after 3 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human NCI-H929 cells assessed as changes in survivin protein level at 30 uM after 3 hrs by immunoblotting 15958589
RPM18226 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human RPM18226 cells at 30 uM after 3 hrs using TUNEL staining by flow cytometry 15958589
NCI-H929 Apoptosis assay 30 uM 1.5 hrs Induction of apoptosis in human NCI-H929 cells assessed as reduction of RNA polymerase 2 phosphoserine 2 level at 30 uM after 1.5 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 1.5 hrs Induction of apoptosis in human NCI-H929 cells assessed as dephosphorylation of pRb at S249/T252 at 30 uM after 1.5 hrs by immunoblotting 15958589
NCI-H929 Cytotoxicity assay 20 to 30 uM 16 hrs Cytotoxicity against human NCI-H929 cells assessed as reduction of cell viability at 20 to 30 uM treated for 16 hrs followed by washout measured after total 72 hrs growth period alamar blue assay relative to control 15958589
NCI-H929 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human NCI-H929 cells assessed as reduction of Mcl-1 protein level at 30 uM after 3 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human NCI-H929 cells assessed as changes in Bcl-2 protein level at 30 uM after 3 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 3 hrs Induction of apoptosis in human NCI-H929 cells at 30 uM after 3 hrs using TUNEL staining by flow cytometry 15958589
NCI-H929 Apoptosis assay 30 uM 1.5 hrs Induction of apoptosis in human NCI-H929 cells assessed as reduction of RNA polymerase 2 phosphoserine 5 level at 30 uM after 1.5 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 1.5 hrs Induction of apoptosis in human NCI-H929 cells assessed as reduction of Hdm2 level at 30 uM after 1.5 hrs by immunoblotting 15958589
NCI-H929 Apoptosis assay 30 uM 1.5 hrs Induction of apoptosis in human NCI-H929 cells assessed as increase of p53 accumulation at 30 uM after 1.5 hrs by immunoblotting 15958589
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 21080703
HCT116 Function assay 30 to 40 umol/L 24 hrs Inhibition of cyclin A in human HCT116 cells assessed as decrease in protein level at 30 to 40 umol/L after 24 hrs by immunoblotting analysis 21080703
HCT116 Function assay 30 to 40 umol/L 24 hrs Inhibition of cyclin B in human HCT116 cells assessed as decrease in protein level at 30 to 40 umol/L after 24 hrs by immunoblotting analysis 21080703
HCT116 Function assay 30 to 40 umol/L 24 hrs Inhibition of cyclin D1 in human HCT116 cells assessed as decrease in protein level at 30 to 40 umol/L after 24 hrs by immunoblotting analysis 21080703
HCT116 Function assay 30 to 40 umol/L 24 hrs Inhibition of CDK2 in human HCT116 cells assessed as decrease in protein level at 30 to 40 umol/L after 24 hrs by immunoblotting analysis 21080703
HT-29 Function assay 2.5 to 40 uM 24 hrs Inhibition of retinoblastoma protein in human HT-29 cells assessed as reduction of cyclin A level at 2.5 to 40 uM after 24 hrs by immunoblotting 21417417
MCF7 Cell cycle assay 24 hrs Cell cycle arrest in human MCF7 cells assessed as accumulation at G2/M phase after 24 hrs using propidium iodide and BrdU staining by flow cytometry 21417417
RPMI8226 Cell cycle assay 24 hrs Cell cycle arrest in human RPMI8226 cells assessed as accumulation at G2/M phase after 24 hrs using propidium iodide and BrdU staining by flow cytometry 21417417
MCF7 Cell cycle assay 24 hrs Cell cycle arrest in human MCF7 cells assessed as decrease in S phase cell population after 24 hrs using propidium iodide and BrdU staining by flow cytometry 21417417
MCF7 Cell cycle assay 24 hrs Cell cycle arrest in human MCF7 cells assessed as accumulation at sub-G1 phase after 24 hrs using propidium iodide and BrdU staining by flow cytometry 21417417
RPMI8226 Cell cycle assay 24 hrs Cell cycle arrest in human RPMI8226 cells assessed as accumulation at sub-G1 phase after 24 hrs using propidium iodide and BrdU staining by flow cytometry 21417417
MCF7 Cell cycle assay 80 uM 24 hrs Cell cycle arrest in human MCF7 cells assessed as reduction of actively replicating DNA level at 80 uM after 24 hrs using propidium iodide and BrdU staining by flow cytometry 21417417
MCF7 Function assay 20 uM 24 hrs Induction of p53-dependent transcriptional activity in human MCF7 cells assessed as increase of p21 WAF1 level at 20 uM after 24 hrs by immunofluorescence assay 21417417
RPMI8226 Cell cycle assay 24 hrs Cell cycle arrest in human RPMI8226 cells assessed as decrease in S phase cell population after 24 hrs using propidium iodide and BrdU staining by flow cytometry 21417417
RPMI8226 Cell cycle assay 80 uM 24 hrs Cell cycle arrest in human RPMI8226 cells assessed as reduction of actively replicating DNA level at 80 uM after 24 hrs using propidium iodide and BrdU staining by flow cytometry 21417417
A549 Apoptosis assay 2 uM 48 hrs Induction of apoptosis in human A549 cells assessed as DNA fragmentation at 2 uM after 48 hrs by agarose gel electrophoresis 23623491
Sf9 Function assay 10 mins Inhibition of His-6-tagged recombinant human CDK2/cyclinE expressed in baculovirus-infected sf9 cells using histone H1 as substrate after 10 mins by liquid scintillation counting in presence of [gamma-32P]ATP, IC50 = 0.1 μM. 24417566
BJ Function assay 10 uM 10 days Suppression of senescence in human BJ cells assessed as increase in cell number at 10 uM after 10 days by senescence reversal assay 24681986
BJ Function assay 10 uM 10 days Inhibition of ataxia telangiectasia-mutated in human BJ cells assessed as increase in cell number at 10 uM after 10 days by senescence reversal assay 24681986
MCF7 Function assay 10 uM 10 mins Sensitization of infrared-induced DNA damage in human MCF7 cells assessed as reduction in colony formation at 10 uM pretreated for 10 mins followed by irradiation for 4 hrs measured after 10 days by crystal violet staining analysis 26851505
Caco2 Cell cycle assay Cell cycle arrest in human Caco2 cells assessed as accumulation at G1/S phase by Hoechst staining based fluorescence assay 28214231
HaCaT Cell cycle assay Cell cycle arrest in human HaCaT cells assessed as accumulation at G1/S phase by Hoechst staining based fluorescence assay 28214231
HuH7 Cell cycle assay Cell cycle arrest in human HuH7 cells assessed as accumulation at G1/S phase by Hoechst staining based fluorescence assay 28214231
PC3 Cell cycle assay Cell cycle arrest in human PC3 cells assessed as accumulation at G2/M phase by Hoechst staining based fluorescence assay 28214231
MDA-MB-231 Cell cycle assay Cell cycle arrest in human MDA-MB-231 cells assessed as accumulation at G1/S phase by Hoechst staining based fluorescence assay 28214231
HCT116 Cell cycle assay Cell cycle arrest in human HCT116 cells assessed as accumulation at G1/S phase by Hoechst staining based fluorescence assay 28214231
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 28557430
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells 29435139
DAOY qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells 29435139
BT-37 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells 29435139
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells 29435139
LAN-5 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 30199702
Click to View More Cell Line Experimental Data

Biological Activity

Description Roscovitine is a potent and selective CDK inhibitor for Cdc2, CDK2 and CDK5 with IC50 of 0.65 μM, 0.7 μM and 0.16 μM in cell-free assays. It shows little effect on CDK4/6. Phase 2.
Targets
CDK5/p35 [1]
(Cell-free assay)		 Cdc2/CyclinB [1]
(Cell-free assay)		 CDK2/CyclinA [1]
(Cell-free assay)		 CDK2/CyclinE [1]
(Cell-free assay)		 ERK2 [1]
(Cell-free assay)
0.16 μM 0.65 μM 0.7 μM 0.7 μM 14 μM
In vitro
In vitro

Roscovitine displays high efficiency and high selectivity towards some cyclin-dependent kinases with IC50 of 0.65, 0.7, 0.7 and 0.16 μM for cdc2/cyclin B, cdk2/cyclin A, cdk2/cyclin E and cdk5/p53, respectively. [1] Roscovitine reversibly inhibits the prophaselmetaphase transition in the micromolar range of starfish oocytes and sea urchin embryos, inhibits in vitro M-phase-promoting factor activity and in vitro DNA synthesis in Xenopus egg extracts, and suppresses the proliferation of mammalian cell lines with an average IC50 of 16 μM. [1] In mesangial cells, Roscovitine results in a dose-dependent reduction of CDK2 activity that at concentrations of 7.5, 12.5 and 25 mM, Roscovitine causes a 25, 50% and 100% decrease in CDK2 activity, respectively. [2] A recent study shows that Roscovitine inhibits cdk5 kinase activity, cell proliferation, multicellular development, and cdk5 nuclear translocation in Dictyostelium discoideum, without affecting the expression of cdk5 protein during axenic growth. [3]
Kinase Assay Enzymes
Kinases activities are assayed at 30 °C in buffer C. Blank values are subtracted from the data and activities calculated as molar amount of phosphate incorporated in protein acceptor during a 10-minute incubation. Controls are performed with appropriate dilutions of DMSO. In a few cases, phosphorylation of the substrate is assessed by autoradiography after SDS/PAGE. p34cdc2/cyclin B is purified from M-phase starfish (M. glacialis) oocytes by affinity chromatography. It is assayed with 1 mg histone Hl/mL, in the presence of 15 μM [γ-32P]ATP (3000 Ci/mmol; 1 mCi/mL) in a final volume of 30 μL. After a 10-minute incubation at 30 °C, 25-μL aliquots of supernatant are spotted onto pieces of Whatman P81 phosphocellulose paper, and, after 20 seconds, the filters are washed five times (for at least 5 minutes each time) in a solution of 10mL phosphoric acid/L water. The wet filters are transferred into 6-mL plastic scintillation vials, 5 mL ACS scintillation fluid is added and the radioactivity measured in a Packard counter. The kinase activity is expressed as molar amount of phosphate incorporated in histone H1 during a 10-minutes incubation or as a percentage of maximal activity. p33cdk2/cyclin A and p33cdk2/cyclinE are reconstituted from extracts of sf9 insect cells infected with various baculoviruses. Cyclins A and E are fusion proteins with glutathione S-transferase and the complexes are purified on glutathione-agarose beads. Kinase activities are assayed with 1 mg/mL histone H1, in the presence of 15 μM [γ-32P]ATP, during 10 minutes, in a final volume of 30 μL, as described for the p34cdc2/cyclin B kinase. p33cdk5/p35 is purified from bovine brain, excluding the Mono S-chromatographic step. The active fractions from the Superose 12 column are pooled and concentrated to a final concentration of approximately 25 μg enzyme/mL. The kinase is assayed with 1 mg/mL histone HI in the presence of 15 μM [γ-32P]ATP, during 10 minutes in a final volume of 30 μL, as described for the p34cdc2/cyclin B kinase. p33cdk5/cyclin D1 is obtained from insect cell lysates. Cdk4 is a fusion protein with glutathione-S-transferase and the active complex is purified on glutathione-agarose beads. Its kinase activity is assayed with purified retinoblastoma protein (complexed with glutathione-S-transferase) in the presence of 15 μM [γ-32P]ATP, in a final volume of 30 μL. After a 15-minute incubation, 30 μL Laemmli sample buffer is added. The phosphorylated substrate is resolved by 10 % SDS/PAGE and analysed by autoradiography by overnight exposure to Hyperfilm MP and densitometry. p33cdk4/cyclinD 2 is obtained from insect cell lysates. It is assayed with purified retinoblastoma protein (complexed with glutathione-S-transferase) in the presence of 15 μM [γ-32P]ATP in a final volume of 30 μL. After a 30-minute incubation, 30 μL Laemmli sample buffer is added. The phosphorylated substrate is resolved by 10% SDS/PAGE and analysed by autoradiography by overnight exposure to Hyperfilm MP and densitometry. MAP kinase erkl (tagged with glutathione-S-transferase), is expressed in bacteria, purified on glutathione-agarose beads and assayed with 1 mg myelin basic protein/ml in the presence of 15 μM [γ-32P]ATP as described above for the p34cdc2cyclin B kinase. His-tagged erkl and erk2 are activated in vitro by mitogen-activated protein kinase kinase, purified (Ni-affinity and Mono Q) and assayed as described above during 10 minutes in a final volume of 30 μL. The catalytic subunit of cAMP-dependent protein kinase, purified from bovine heart, is assayed with 1 mg histone Hl/ml, in the presence of 15 μM [γ-32P]ATP as described for the p34cdc2/cyclin B kinase. cGMP-dependent protein kinase, purified to homogeneity from bovine tracheal smooth muscle, is assayed with 1 mg histone Hl/mL, in the presence of 15 μM [γ-32P]ATP as described for the p34cdc2/cyclin B kinase. Casein kinase 2 is isolated from rat liver cytosol and assayed with 1 mg casein/mL and 15 μM [γ-32P]ATP. The substrate is spotted on Whatmann 3MM filters and washed with 10% (mass/vol.) trichloroacetic acid. Myosin light chain kinase, purified from chicken gizzard is assayed in the presence of 100 nM calmodulin, 100 μM CaCl2, 50 mM Hepes, 5 mM MgCI,, 1 mM dithiothreitol and 0.1 mg BSA/ml at pH 7.5 using a synthetic peptide based on the smooth-muscle myosin light-chain phosphorylation site and in the presence of 15 μM [γ-32P]ATP, in a final volume of 50 μL. Incorporation of radioactive phosphate is monitored on phosphocellulose filters as described above. ASK-γ, a plant homologue of GSK-3, is expressed as a glutathione-S-transferase fusion protein in Escherichia coli and purified on glutathione-agarose. ASK-γ kinase is assayed, for 10 minutes at 30 °C, with 5 μg myelin basic protein, in the presence of 15 μM [γ-32P]ATP in a final volume of 30 μL. The phosphorylated myelin basic protein is recovered on Whatman P81 phosphocellulose paper as described for the p34cdc2/cyclin B kinase. Insulin receptor tyrosine kinase domain (CIRK-41) is overexpressed in a baculovirus system and purified to homogeneity. Its kinase activity is assayed, for 10 minutes at 30 °C, with 5 μg Raytide, in the presence of 15 μM [γ-32P]ATP, in a final volume of 30 μL. The phosphorylated Raytide is recovered on Whatman P81 phosphocellulose paper as described for the p34cdc2/cyclin B kinase. c-src kinase is purified from infected Sf9 cells. The v-abl kinase is expressed in E. coli and affinity purified on IgG Affigel 10. Both kinases are assayed for 10 minutes at 30 °C, with 5 μg Raytide, in the presence of 15 μM [γ-32P]ATP, in a final volume of 30 μL. The phosphorylated Raytide is recovered on Whatman P81 phosphocellulose paper as described for the p34cdc2/cyclin B kinase.
Cell Research Cell lines Leukemia, non-small cell lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer
Concentrations 0.01 - 100 μM
Incubation Time 48 hours
Method

60 human tumour cell lines comprising nine tumor types are cultured for 24 hours prior to a 48-hour continuous exposure to 0.01-100 μM roscovitine. A sulforhodaminine B protein assay is used to estimate the cytotoxicity.
Experimental Result Images Methods Biomarkers Images PMID
Western blot pT231-tau / pS202-tau / tau p-Rb / p-CDK2 / CDK2 / Cyclin D1 / Cyclin A2 / ERα / ERβ/ AIB1 / PELP1 30915013
Immunofluorescence CDK1 / Smek2 / FUBP1 / Cdc20 E2F1 / FASN / Bmi1 / Cyclin D2 / CDK2 / CDK4 24534090
Growth inhibition assay Cell viability 29996940
In Vivo
In vivo

Roscovitine, at a dose of 50 mg/kg, significantly inhibits growth of The Ewing's sarcoma family of tumors (ESFT) xenografts. [4] Roscovitine enhances the antitumor effect of doxorubicin without increased toxicity with a mechanism that involves cell cycle arrest rather than apoptosis in nude mice bearing established MCF7 xenografts. [5]
Animal Research Animal Models A4573 cells are injected s.c. into the right posterior flank of CD1 nu/nu mice.
Dosages ≤50 mg/kg
Administration Administered via i.p.
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT02649751 Terminated
Cystic Fibrosis
University Hospital Brest|ManRos Therapeutics|Cyclacel Pharmaceuticals Inc.
 February 22 2016 Phase 2

Chemical Information & Solubility

Molecular Weight 354.45 Formula

C19H26N6O
CAS No. 186692-46-6 SDF Download Roscovitine SDF
Smiles CCC(CO)NC1=NC(=C2C(=N1)N(C=N2)C(C)C)NCC3=CC=CC=C3
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 71 mg/mL ( (200.31 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 71 mg/mL

Water : Insoluble


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Answer:
S1153 in 1% DMSO+10% Tween 80+20% N-N-dimethylacetamide+PEG 400 is a clear solution which is okay for injection. And S1153 in 1% DMSO+30% polyethylene glycol+1% Tween 80 at 30mg/ml is a suspension, which is fine for oral gavage.

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