CYT387

 

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Biological Activity

Description	CYT387 is an ATP-competitive inhibitor of JAK1/JAK2 with IC50 of 11 nM/18 nM, ~10-fold selectivity versus JAK3. Phase 1/2.
Targets	JAK1	JAK2	JAK3
IC50	11 nM	18 nM	155 nM [1]
In vitro	CYT387 inhibits the proliferation of parental Ba/F3 cells (Ba/F3-wt) stimulated by IL-3 with IC50 of 1400 nM. Furthermore, CYT387 also causes the inhibition of cell proliferation in cell lines constitutively activated by JAK2 or MPL signaling, including Ba/F3-MPLW515L cells, CHRF-288-11 cells and Ba/F3-TEL-JAK2 cells with IC50 of 200 nM, 1 nM and 700 nM, respectively. In addition, CYT387 has been shown to inhibit erythroid colony growth in vitro from JAK2V617F-positive PV patients with similar potency with IC50 of 2μ-4 μM. [1] A recent study shows that CYT387 inhibits PI3K/AKT and Ras/MAPK signaling induced by IL-6 and IGF-1. Moreover, CYT387 induces apoptosis as a single agent and synergizes with the conventional anti-MM therapies bortezomib and melphalan in primary multiple myeloma (MM) cells. [2]
In vivo	In a murine MPN model, CYT387 normalizes white cell counts, hematocrit, spleen size, and restores physiologic levels of inflammatory cytokines. [3]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Cell-free kinase activity assays	Glutathione-S-transferase (GST)-tagged JAK kinase domains expressed in insect cells are purified before use in a peptide substrate phosphorylation assay. Assays are carried out in 384-well optiplates using an Alphascreen Protein Tyrosine Kinase P100 detection kit and a PerkinElmer Fusion Alpha instrument.

Cell Assay: ^[1]

Cell lines	Ba/F3, Ba/F3-JAK2V617F and Ba/F3-MPLW515L cells
Concentrations	0 to 10 μM
Incubation Time	72 hours
Method	Ba/F3 cells expressing JAK2V617F (Ba/F3-JAK2V617F) and MPLW515L (Ba/F3-MPLW515L) mutants, as well as CHRF-288-11 (JAK2T875N) and CMK (JAK3A572V) cells are used. The TEL/JAK2 and TEL/JAK3 fusions are generated and introduced into Ba/F3 murine cells. The TEL/JAK2- or TEL/JAK3-transfected cells are cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal calf serum (FCS). Ba/F3 wild-type cells are cultured in RPMI containing 10% FCS supplemented with 5 ng/mL murine IL-3. Proliferation is measured using the Alamar Blue assay after incubating for 72 hours at 37 °C with 5% CO2

Animal Study: ^[3]

Animal Models	Balb/c mice are transplanted with bone marrow transduced with a JAK2V617F retrovirus.
Dosages	≤50 mg/kg
Administration	Administered via p.o.
Solubility	30% PEG400/0.5% Tween80/5% propylene glycol, 30 mg/mL

1

References

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

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT00935987	Completed	Primary Myelofibrosis|Post-Polycythemia Vera Myelofibrosis|Post-Essential Thrombocythemia Myelofibrosis	Gilead Sciences	2009-11	Phase 1|Phase 2
NCT01236638	Active, not recruiting	Primary Myelofibrosis|Post-Polycythemia Vera Myelofibrosis|Post-Essential Thrombocythemia Myelofibrosis	Gilead Sciences	2010-11	Phase 2
NCT01423058	Active, not recruiting	Primary Myelofibrosis|Post-Polycythemia Vera|Post-Essential Thrombocythemia Myelofibrosis	Gilead Sciences	2011-08	Phase 1|Phase 2

Chemical Information

Download CYT387 SDF

Chemical Name	N-(cyanomethyl)-4-(2-(4-morpholinophenylamino)pyrimidin-4-yl)benzamide

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.41446	4.1446	8.2892	12.4338

Customer Reviews (1)

Click to enlarge	Rating
	Source	Blood, 2012, 120(19):4093-103. CYT387 purchased from Selleck
	Method	Flow cytometric analysis
	Cell Lines	WT pan T cells
	Concentrations	1 uM
	Incubation Time
	Results	CYT387 dramatically reduced the expression of CXCR3 in activated WT T cells.

Product Citations (1)

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  IFNγR signaling mediates alloreactive T cell trafficking and GvHD. [Choi J, et al. Blood 2012;120(19):4093-103]