Danusertib (PHA-739358)

 

Biological Activity

Description	Danusertib (PHA-739358) is an Aurora kinase inhibitor for Aurora A/B/C with IC50 of 13 nM/79 nM/61 nM, modestly potent to Abl, TrkA, c-RET and FGFR1, and less potent to Lck, VEGFR2/3, c-Kit, CDK2, etc. Phase 2.
Targets	Aurora A/B/C	Bcr-Abl	c-RET	FGFR	TrkA
IC50	13 nM/79 nM/61 nM	25nM	31 nM	47 nM	30 nM [2]
In vitro	Danusertib inhibits the activities of other kinases such as FGFR1, Abl, Ret and Trka, with IC50 of 47 nM, 25 nM, 31 nM and 31 nM, respectively. In a cell assay, after treatment of wild-type and p53-deficient MEFs with Danusertib, the wild-type cells undergo an arrest in mitosis (4N) that is maintained for up to 48 h. The p53-deficient cells on the other hand do not arrest at the 4N DNA stage, but continues with additional rounds of DNA synthesis to become >8N. Treatment with Danusertib results in an increase in p53 protein levels and an associated increase in p21 protein, which is known to be transcriptionally regulated by p53. [1] Increasing concentrations of Danusertib produces a dose-dependent reduction of cell growth after 48 hours in BCR-ABL-positive (K562, BV173) and BCR-ABL-negative (HL60) cells. [3]
In vivo	Administration of 25 mg/kg Danusertib (b.d. i.v.) to HL-60 xenograft rats results in 75% inhibition of tumor growth with complete regression in one animal. Danusertib results in biomarker modulation accompanied by inhibition of tumor growth. This is compatible with an expected mechanism of action of aurora kinase inhibition.[1] Danusertib significantly inhibits proliferation of K562 cells and virtually suppressed tumor growth during the 10-day treatment period. [3]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Biochemical kinase Assays	The Km values for ATP and the specific substrate are initially determined, and each assay is then run at optimized ATP (2Km) and substrate (5Km) concentrations. This setting enabled direct comparison of IC50 values of Danusertib across the applied kinase selectivity screening panel for the evaluation of the selectivity profile.

Cell Assay: ^[3]

Cell lines	CD34+ cells
Concentrations	5 μM
Incubation Time	5 days
Method	For short-term expansion assays, 1 × 103 CD34+ cells are plated in triplicates in 96-well plates containing 100 μL of serum-free medium per well supplemented with human stem-cell factor (100 ng/mL), human Flt-3 Ligand (100 ng/mL), human thrombopoietin (50 ng/mL), human interleukin-3 and -6 (IL-3 and IL-6, respectively, both 20 ng/mL), and granulocyte colony-stimulating factor (20 ng/mL) along with Danusertib at the indicated concentrations. After 5 days, an additional 100 μL of cytokine and Danusertib containing medium are added. Cell numbers within each individual well are estimated on days 3, 6, and 9 or on days 3, 6, and 12 for healthy donor samples.

Animal Study: ^[3]

Animal Models	Female SCID mice
Dosages	15 mg/kg
Administration	Intraperitoneally
Solubility	1% DMSO/30% polyethylene glycol/1% Tween 80, 30 mg/mL

1

References

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

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT00335868	Active, not recruiting	Leukemia	National Cancer Institute (NCI)|Jonsson Comprehensive Cancer Center	2007-03	Phase 2
NCT00766324	Completed	Metastatic Hormone Refractory Prostate Cancer	Nerviano Medical Sciences	2007-09	Phase 2
NCT00872300	Terminated	Multiple Myeloma	Nerviano Medical Sciences	2008-10	Phase 2

Chemical Information

Download Danusertib (PHA-739358) SDF

Chemical Name	(R)-N-(5-(2-methoxy-2-phenylacetyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.47455	4.7455	9.491	14.2365

Customer Reviews (2)

Click to enlarge	Rating
	Source	Dr. Yong-Weon Yi from Georgetown University Medical Center.Danusertib (PHA-739358) purchased from Selleck
	Method	MTT assays
	Cell Lines	MDA-MB-231 cells
	Concentrations	0.01-10 μM
	Incubation Time	72 h
	Results	PHA-739358 decreased the viability of MDA-MB-231 cells in a dose-dependent manner.

Product Citations (5)

• 
  

  Targeting Sonic Hedgehog-Associated Medulloblastoma through Inhibition of Aurora and Polo-like Kinases. [Markant SL, et al. Cancer Res 2013;73(20):6310-22]

  
  PubMed: 24067506


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  Kinome-wide siRNA screening identifies molecular targets mediating the sensitivity of pancreatic cancer cells to Aurora kinase inhibitors. [Xie L, et al. Biochem Pharmacol 2012;83(4):452-61]

  
  PubMed: 22100984


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  Preclinical characterization of ABT-348, a kinase inhibitor targeting the aurora, vascular endothelial growth factor receptor/platelet-derived growth factor receptor, and Src kinase families. [Glaser KB, et al. J Pharmacol Exp Ther 2012;343(3):617-27]

  
  PubMed: 22935731