GW2580 (SC203877)

c-FMS (IC50 = 60 nM)

GW2580 completely inhibits human cFMS kinase in vitro at a concentration of 0.06 μM. With an IC50 of 0.33, 13.5, 0.47, and 12 μM, respectively, GW2580 suppresses the growth of myeloid tumor cells stimulated by CSF-1, serum-stimulated NSO myeloid tumor cells, freshly isolated human monocytes stimulated by CSF-1, and human umbilical vein vascular endothelial cells stimulated by VEGF. A single microm Complete inhibition of bone degradation in cultures of human osteoclasts, rat calvaria, and rat fetal long bone is observed with GW2580. It also completely inhibits the growth of mouse M-NFS-60 myeloid cells and human monocytes induced by CSF-1.[1] With an IC50 of about 10 nM, GW2580 suppresses CSF1R phosphorylation in RAW264.7 murine macrophages stimulated with 10 ng/mL. With an IC50 of 0.88 μM, GW2580 likewise suppresses TRKA activity [2][3]

GW2580 (dosed orally at 40 mg/kg 0.5 h before the CSF-1-priming dose) suppresses the 63% increase in TNF-α production in mice induced by LPS when exogenous CSF-1 is added. GW2580 completely prevents CSF-1 from priming mice for enhanced IL-6 production when it is administered to them prior to CSF-1 priming. The peritoneal cavity's M-NFS-60 tumor cells that are dependent on CSF-1 are totally inhibited from growing by GW2580 (80 mg/kg p.o.). When taken orally twice a day the week prior to and for four days following thioglycolate injection, GW2580 (80 mg/kg) reduces (by 45%) the amount of macrophages that accumulate in the peritoneal cavity.[1] GW2580 (50 mg/kg), administered twice daily from days 0 to 21, 7 to 21, or 14 to 21, inhibits the breakdown of joint connective tissue and bone in a 21-day adjuvant arthritis model.[3] By preventing the tumor from recruiting myeloid cells from peripheral blood, Gw2580 (160 mg/kg) causes a more than two-fold decrease in total CD45⁺ CD11b⁺ myeloid cells, CD11b⁺ F4/80⁺ TAMs, and CD11b⁺ Gr-1⁺ MDSCs in implanted 3LL lung tumors. Treatment with GW2580 (80 mg/kg) can reduce tumor vascular density (CD31 staining) and the expression of Vegf-a (by 35%) and Mmp9 (by 70%). Combination therapy involving GW2580 and an anti-VEGFR-2 antibody reduces tumor growth in a synergistic manner. While DC101 by itself inhibits tumor growth by 35%, when combined with GW2580, there appears to be a synergistic reduction in tumor growth of about 70%.[2]

After 90 minutes at room temperature, 10 μM enzyme, 100 μM ATP, and 5 mM MgCl₂ are incubated in 50 mM Tris HCL to activate the enzyme through autophosphorylation. Round-bottom polystyrene 96-well plates on a Biomek 2000 are used to conduct 45 μL enzyme reactions. In each well, 30 mL of a 1.5-substrate reaction mix containing 50 mM Mops (3-[N-Morpholino]propanesulfonic acid), pH 7.5, 15 mM MgCl₂, 6 M peptide substrate, and biotin-EAIYAPFAKKK-NH2 is added, either alone or in 1 mL of DMSO. 0.5 μCi (1 Ci = 37 GBq) [³³P-γ] ATP, 75 mM NaCl, 10 μM ATP, and 7.5 mM DTT are needed for each assay. The final enzyme concentration is 20 nM, and the reaction is started by adding 15 μL of diluted enzyme solution. Background is determined by adding EDTA to the control wells. A 96-well phosphocellulose filter plate is prewet with 100 μL of 0.5% phosphoric acid, and 75 μL of the reaction is transferred to it after it has been allowed to proceed for 40 minutes and stopped by adding an equal volume of the acid. Following three rounds of washing with the phosphoric acid solution and filtering on a Millipore filter-plate vacuum manifold, 40 μL of scintillation solution is added. In a Packard Topcount NXT scintillation counter, the plates are sealed and tallied.

The cells are spun down and diluted media-infused with 2× 10⁶ cells/ml for 24 hours prior to the commencement of the cell growth assay. M-NSF60 cell depleted medium is devoid of MCSF. The following day, GW2580 at 10 mM in DMSO is serially diluted to produce a 10-point concentration curve, starting at 20 μM and 0.2% DMSO in medium containing 10% serum. After being resuspended in medium, the M-NFS-60 cells are added to 0.5× 10⁶ cells/mL along with 10% serum and 20 ng/mL mouse MCSF. Each well is filled with 50 μL of inhibitor-containing cells, and then, after three days, 10 μL of WST-1 reagent is added to each well. Growth is determined by measuring the difference between wells with full medium and wells with depleted medium after a 4-hour incubation period. The absorbance is measured at 440 nm.

Mouse myeloid carcinoma xenografts M-NFS-60
80 mg/kg
Orally twice a day

[1]. Inhibition of colony-stimulating-factor-1 signaling in vivo with the orally bioavailable cFMS kinase inhibitor GW2580. Proc Natl Acad Sci U S A. 2005 Nov 1;102(44):16078-83.

[2]. Targeting distinct tumor-infiltrating myeloid cells by inhibiting CSF-1 receptor: combating tumor evasion of antiangiogenic therapy. Blood. 2010 Feb 18;115(7):1461-71

[3]. Effects of the cFMS kinase inhibitor 5-(3-methoxy-4-((4-methoxybenzyl)oxy)benzyl)pyrimidine-2,4-diamine (GW2580) in normal and arthritic rats. J Pharmacol Exp Ther. 2008 Jul;326(1):41-50.

C20H22N4O3

366.41

366.17

C, 65.56; H, 6.05; N, 15.29; O, 13.10

870483-87-7

white solid powder

COC1=CC=C(C=C1)COC2=C(C=C(C=C2)CC3=CN=C(N=C3N)N)OC

MYQAUKPBNJWPIE-UHFFFAOYSA-N

InChI=1S/C20H22N4O3/c1-25-16-6-3-13(4-7-16)12-27-17-8-5-14(10-18(17)26-2)9-15-11-23-20(22)24-19(15)21/h3-8,10-11H,9,12H2,1-2H3,(H4,21,22,23,24)

5-[[3-methoxy-4-[(4-methoxyphenyl)methoxy]phenyl]methyl]pyrimidine-2,4-diamine

GW 2580; SC-203877; SC 203877; GW 2580; GW-2580; SC203877

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)