Cyt387

• Catalog No.: S2219
• CAS Number: 1056634-68-4
• M. F.: C23H22N6O2
• M. W.: 414.45978
• Storage: -20°C

SYNONYMS

• IUPAC name: N-(cyanomethyl)-4-(2-{[4-(morpholin-4-yl)phenyl]amino}pyrimidin-4-yl)benzamide
• IUPAC Traditional name: N-(cyanomethyl)-4-(2-{[4-(morpholin-4-yl)phenyl]amino}pyrimidin-4-yl)benzamide
• Synonyms: CYT 11387; Cyt-387

DATABASE IDS

• CAS Number: 1056634-68-4

PROPERTIES

• Target: JAK
• Salt Data: Free Base
• Storage Condition: -20°C

DETAILS

Description (English)

Research Area

• Description: Myeloproliferative disorders

Biological Activity

• Description: CYT387 is an ATP-competitive inhibitor of JAK1 and JAK2 with IC50 of 11 nM and 18 nM, respectively.
• 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]
• Clinical Trials: Cyt387 is currently in Phase I/II clinical trials in patients with primary myelofibrosis or post-polycythemia vera or post-essential thrombocythemia myelofibrosis.

Protocol

• Protocol: 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.
• Protocol: Cell Assay ^[1]
• Cell Lines: Ba/F3, Ba/F3-JAK2V617F and Ba/F3-MPLW515L cells
• Concentrations: 0 to 10 μM
• Incubation Time: 72 hours
• Methods: 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.
• Protocol: Animal Study ^[3]
• Animal Models: Balb/c mice are transplanted with bone marrow transduced with a JAK2V617F retrovirus.
• Formulation: CYT387 is dissolved in NMP (120 mg/mL final; 1-methyl-2-pyrrolidinone, Chromasolv Plus). Subsequently, the CYT387/NMP mix is diluted with 0.14 M Captisol to a concentration of 6 mg/mL and further diluted with 0.1M Captisol to a final concentration of 4 mg/mL.
• Doses: ≤50 mg/kg
• Administration: Administered via p.o.

References

• References: [1] Pardanani A, et al. Leukemia, 2009, 23(8), 1441-1445.
• References: [2] Monaghan KA, et al. Leukemia, 2011, 25(12), 1891-1899.
• References: [3] Tyner JW, et al. Blood, 2010, 115(25), 5232-5240.

REFERENCES

• Pardanani A, et al. Leukemia, 2009, 23(8), 1441-1445.
• Monaghan KA, et al. Leukemia, 2011, 25(12), 1891-1899.
• Tyner JW, et al. Blood, 2010, 115(25), 5232-5240.