Dinaciclib (SCH727965)

 

Biological Activity

Description	Dinaciclib (SCH727965) is a novel and potent CDK inhibitor for CDK2, CDK5, CDK1 and CDK9 with IC50 of 1 nM, 1 nM, 3 nM and 4 nM, respectively. It also blocks thymidine (dThd) DNA incorporation. Phase 3.
Targets	CDK2	CDK5	CDK1	CDK9
IC50	1 nM	1 nM	3 nM	4 nM [1]
In vitro	Dinaciclib is also a potent DNA replication inhibitor that blocks thymidine (dThd) DNA incorporation in A2780 cells with IC50 of 4 nM. Dinaciclib strongly suppresses phosphorylation of Rb on Ser 807/811 at concentrations >6.25 nM, which is in agreement with the observation that 4 nM concentrations are required for 50% inhibition of dThd DNA incorporation in the same cell model. Significantly, complete suppression of Rb phosphorylation is correlated with the onset of apoptosis, as indicated by the appearance of the p85 PARP cleavage product in cells exposed to >6.25 nM Dinaciclib. Dinaciclib is active against a broad spectrum of human tumor cell lines.[1] Addition of Dinaciclib during hydroxyurea exposure also suppresses accumulation of γ-H2AX, in a dose-dependent manner. [2] Dinaciclib inhibits melanoma cell proliferation, and drives melanoma cells into massive apoptosis. [3] Dinaciclib induces the apoptosis of several osteosarcoma cell lines including those resistant to doxorubicin and dasatinib. Dinaciclib attenuates the phosphorylation of RNAP II at serine 2 and the phosphorylation of the CDK inhibitor p27Kip1 at threonine 187. Reductions in phosphorylation activity occurrs at 12 - 40 nM Dinaciclib (4 to 16 hours post-Dinaciclib addition). Dinaciclib also reduces the phosphorylation of Rb at serine 807/811. Dinaciclib induces the apoptosis of mock- and p53-depleted U2OS cells to a similar extent.[4]
In vivo	Dinaciclib i.p. administration at 8, 16, 32, and 48 mg/kg daily for 10 days results in tumor inhibition by 70%, 70%, 89%, and 96%, respectively. Dinaciclib MED (minimum effective dose) appears to be <8 mg/kg. Dinaciclib is well tolerated, and the maximum body weight loss in the highest dosage group is 5%. Dinaciclib has dose-dependent antitumor activity in vivo, and that nearly complete inhibition of tumor growth occurs at a dose level below the MTD (maximum tolerated dose). Dinaciclib has a short plasma half-life in mouse. [1]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Cyclin/CDK kinase assay

Recombinant cyclin/CDK holoenzymes are purified from Sf9 cells engineered to produce baculoviruses that express a specific cyclin or CDK. Cyclin/CDK complexes are typically diluted to a final concentration of 50 μg/mL in a kinase reaction buffer containing 50 mM Tris-HCl (pH 8.0), 10 mM MgCl2, 1 mM DTT, and 0.1 mM sodium orthovanadate. For each kinase reaction, 1 μg of enzyme and 20 μL of a 2-μM substrate solution (a biotinylated peptide derived from histone H1) are mixed and combined with 10 μL of diluted Dinaciclib. The reaction is started by the addition of 50 μL of 2 μM ATP and 0.1 μCi of 33P-ATP. Kinase reactions are incubated for 1 hour at room temperature and are stopped by the addition of 0.1% Triton X-100, 1 mM ATP, 5 mM EDTA, and 5 mg/mL streptavidin-coated SPA beads. SPA beads are captured using a 96-well GF/B filter plate and a Filtermate universal harvester. Beads are washed twice with 2 M NaCl and twice with 2 M NaCl containing 1% phosphoric acid. The signal is then assayed using a TopCount 96-well liquid scintillation counter.

Cell Assay: ^[1]

Cell lines	A2780 cells
Concentrations	0 μM -5 μM
Incubation Time	24 hours
Method	A2780 cells are maintained in DMEM plus 10% fetal bovine serum and passaged twice weekly by detaching the monolayer with trypsin-EDTA. One hundred microliters of A2780 cells (5 × 103 cells) are added per well to a 96-well Cytostar-T plate and incubated for 16 hours to 24 hours at 37 °C. Dinaciclib is serially diluted in complete media plus 2% 14C-labeled dThd. Media are removed from the Cytostar T plate; 200 μL of various Dinaciclib dilutions are added in quadruplicate; and the cells are incubated for 24 hours at 37 °C. Accumulated incorporation of radiolabel is assayed using scintillation proximity and measured on a TopCounTM.

Animal Study: ^[1]

Animal Models	Nude mice bearing A2780 tumors
Dosages	8 mg/kg, 16 mg/kg, 32 mg/kg, and 48 mg/kg
Administration	Administered via i.p.
Solubility	15% Captisol, 6 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01624441	Recruiting	Estrogen Receptor-negative Breast Cancer|HER2-negative Breast Cancer|Male Breast Cancer|Progesterone Receptor-negative Breast Cancer|Recurrent Breast Cancer|Stage IV Breast Cancer|Triple-negative Breast Cancer	National Cancer Institute (NCI)	2012-08	Phase 1
NCT01676753	Recruiting	Malignant Solid Tumour|Triple-negative Breast Cancer	University of California, San Francisco|Jo Chien|Merck	2012-08	Phase 1
NCT01650727	Recruiting	Chronic Lymphocytic Leukemia|Small Lymphoctyic Lymphoma	Merck	2012-10	Phase 1
NCT01783171	Recruiting	Acinar Cell Adenocarcinoma of the Pancreas|Duct Cell Adenocarcinoma of the Pancreas|Recurrent Pancreatic Cancer|Stage III Pancreatic Cancer|Stage IV Pancreatic Cancer	National Cancer Institute (NCI)	2013-01	Phase 1
NCT01711528	Not yet recruiting	Refractory Multiple Myeloma	National Cancer Institute (NCI)	2013-12	Phase 1

Chemical Information

Download Dinaciclib (SCH727965) SDF

Chemical Name

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.39649	3.9649	7.9298	11.8947

Customer Reviews (1)

Click to enlarge	Rating
	Source	Dinaciclib (SCH727965) purchased from Selleck
	Method
	Cell Lines	MEC1/3T3/Riva/OCI-ly8
	Concentrations	0-50 nM
	Incubation Time	24 h
	Results

Product Citations (1)

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  Cyclin-dependent kinase inhibitor dinaciclib interacts with the acetyl-lysine recognition site of bromodomains. [Martin MP, et al. ACS Chem Biol 2013;8(11):2360-5]