LY2784544_DataSheet_MedChemExpress

Inhibitors, Agonists, Screening Libraries

https://www.sodocs.net/doc/1c8515289.html, Data Sheet

BIOLOGICAL ACTIVITY:

LY2784544 is a potent JAK2 inhibitor with IC 50 of 3 nM. LY2784544 also inhibits FLT3, FLT4, FGFR2, TYK2, and TRKB with IC 50 of 4, 25,32, 44, and 95 nM.

IC50 & Target: IC50: 2.52±0.49 nM (JAK2), 19.8±3 nM (JAK1), 48±14.9 nM (JAK3), 4 nM (FLT3), 32 nM (FGFR2)[1]

In Vitro: LY2784544, a potent, selective and ATP–competitive inhibitor of janus kinase 2 (JAK2) tyrosine kinase. LY2784544

effectively inhibits JAK2V617F–driven signaling and cell proliferation in Ba/F3 cells (IC 50=20 and 55 nM, respectively). In

comparison, LY2784544 is much less potent at inhibiting interleukin–3–stimulated wild–type JAK2–mediated signaling and cell proliferation (IC 50=1183 and 1309 nM, respectively). LY2784544 potently inhibits the JAK2V617F signaling (IC 50=20 nM) but,remarkably, shows very minimal activity against the IL–3–activated wild–type JAK2 signaling with an IC 50 of 1183 nM. LY2784544inhibits the proliferation of JAK2V617F–expressing cells (IC 50=55 nM) and is markedly less potent as an inhibitor of the

proliferation of IL–3–stimulated wild–type JAK2 expressing Ba/F3 cells (IC 50=1309 nM). LY2784544 is potent in the cell–based TF–1 JAK2 assay (IC 50=45 nM) and had the desired threshold selectivity in the NK–92 JAK3/JAK1 heterodimer assay (942 nM)[1].In Vivo: LY2784544 effectively inhibits STAT5 phosphorylation in Ba/F3–JAK2V617F–GFP (green fluorescent protein) ascitic

tumor cells (TED 50=12.7 mg/kg) and significantly reduces (P<0.05) Ba/F3–JAK2V617F–GFP tumor burden in the

JAK2V617F–induced MPN model (TED 50=13.7 mg/kg, twice daily)[1].

PROTOCOL (Extracted from published papers and Only for reference)

Cell Assay: LY2784544 is dissolved in DMSO and stored, and then diluted with appropriate medium before use [1].[1]Ba/F3 cells

expressing JAK2V617F are placed in RPMI–1640–containing vehicle (DMSO) or LY2784544 (concentration range, 0.001–20 μM) (1×104cells/96–well). Ba/F3 cells expressing wild–type JAK2 are treated similarly except IL–3 (2 ng/mL) is added. After a 72–hour incubation,cell proliferation is assessed by adding Cell Titer 96 Aqueous One Solution Reagent (20 μL/well). The IC 50 for inhibition of cell proliferation is calculated using the GraphPad Prism 4 software [1].

Animal Administration: LY2784544 is dissolved in DMSO and diluted with PBS or saline [1].[1]Mice [1]

Dose– and time–dependent in vivo inhibition of JAK2V617F signaling is assessed by measuring inhibition of STAT5 phosphorylation in a mouse ascitic tumor model. Ba/F3–JAK2V617F–GFP cells (1×107) are implanted in the intraperitoneal cavity of severe combined

immunodeficiency mice and allowed to develop into ascitic tumors for 7 days. For dose–response studies (six animals/group),LY2784544 is administered once by oral gavage (2.5, 5, 10, 20, 40, or 80 mg/kg), then 30 min later, ascitic tumor cells are collected,fixed, incubated for 2 h with Mouse–anti–pSTAT5 (pY694) Alexa Fluor 647 (1:10 dilution), and analyzed by flow cytometry. Time course studies are performed similarly, except the animals are treated with LY2784544 at 20, 40 or 80 mg/kg and ascitic tumor cells collected at prespecified intervals of 0.25–6 h after dosing. Data are analyzed by the one–way analysis of variance, and Dunnett's test (α=0.05).Dose response data are analyzed with a four–parameter logistic curve–fitting program.Product Name:

LY2784544Cat. No.:

HY-13034CAS No.:

1229236-86-5Molecular Formula:

C 23H 25ClFN 7O Molecular Weight:

469.94Target:

JAK; JAK; JAK; FLT3; FGFR Pathway:

Epigenetics; Stem Cell/Wnt; JAK/STAT Signaling; Protein Tyrosine Kinase/RTK; Protein Tyrosine Kinase/RTK Solubility:

10 mM in DMSO

References:

[1]. Ma L, et al. Discovery and characterization of LY2784544, a small–molecule tyrosine kinase inhibitor of JAK2V617F. Blood Cancer J. 2013, 3, e109.

Caution: Product has not been fully validated for medical applications. For research use only.

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