Table 1

BAY 43-9006 inhibits the RAF/MEK/ERK pathway and receptor tyrosine kinases involved in tumor angiogenesis Embedded Image

IC50 (nmol/L) ± SD (n)*
Biochemical assay
 Raf-16 ± 3 (7)
 BRAF wild-type§22 ± 6 (7)
 V599E BRAF mutant38 ± 9 (4)
 VEGFR-290 ± 15 (4)
 mVEGFR-2 (flk-1)15 ± 6 (4)
 mVEGR-320 ± 6 (3)
 mPDGFR-β57 ± 20 (5)
 Flt-358 ± 20 (3)
 c-KIT68 ± 21 (3)
 FGFR-1580 ± 100 (3)
 ERK-1, MEK-1, EGFR, HER-2, IGFR-1, c-met, PKB, PKA, cdk1/cyclinB, PKCα, PKCγ, pim-1>10,000
Cellular mechanism
 MDA MB 231 MEK phosphorylation (human breast)40 ± 20 (2)
 MDA MB 231 ERK 1/2 phosphorylation (human breast)90 ± 26 (7)
 BxPC-3 ERK 1/2 phosphorylation (human pancreatic)1,200** ± 165 (2)
 LOX ERK 1/2 phosphorylation (human melanoma)880** ± 90 (2)
 VEGFR-2 phosphorylation (human, NIH 3T3 cells)30 ± 21 (3)
 VEGF–ERK 1/2 phosphorylation (human, HUVEC)60** ± 26 (2)
 PDGFR-β phosphorylation (human HAoSMC)80 ± 40 (3)
 mVEGFR-3 phosphorylation (mouse, HEK-293 cells)100 ± 80 (2)
 Flt-3 phosphorylation (human ITD, HEK-293 cells)20 ± 10 (2)
Cellular proliferation
 MDA MB 231 (10% FCS)2,600 ± 810 (3)
 PDGF-BB HAoSMC (0.1% BSA)280 ± 140 (5)
  • * IC50 mean ± SD; (n = number of trials).

  • Kinase assay were carried out as described in Materials and Methods at ATP concentrations at or below Km (1 to 10 μmol/L).

  • Lck activated NH2-terminal–truncated Raf-1.

  • § NH2-terminal–truncated BRAF (wild-type).

  • NH2-terminal V599E-truncated BRAF (mutant).

  • Cellular mechanism assays (RTK autophosphorylation and RAF/MEK/ERK pathway) were performed in 0.1% BSA using phospho-specific antibodies or 4G10 for VEGFR-3 as described in Materials and Methods.

  • ** Activated phospho-ERK 1/2 was quantitated with phospho-ERK 1/2 immunoassay (Bio-Plex; Bio-Rad, Inc.).