Enzastaurin

Athymic nude mice

3-(1-methyl-1H-indol-3-yl)-4-(1-(1-(pyridin-2-ylmethyl)piperidin-4-yl)-1H-indol-3-yl)-1H-pyrrole-2, 5-dione

0.3-4 uM

75 mg/kg twice daily

6 nM, 6 nM, 6 nM, 6 nM, 6 nM, 6 nM

Treatment of xenografts with Enzastaurin and radiation produced greater reductions in density of microvessels than either treatment alone. The decrease in microvessel density corresponded to delayed tumor growth. [3]

Kinase inhibition assays, The inhibition of PKCbetaII, PKCalpha, PKCepsilon, or PKCgamma activity by enzastaurin is determined using a filter plate assay format measuring 33P incorporation into myelin basic protein substrate. Reactions are done in 100 uL reaction volumes in 96-well polystyrene plates with final conditions as follows: 90 mM HEPES (pH 7.5), 0.001% Triton X-100, 4% DMSO, 5 mM MgCl2, 100 uM CaCl2, 0.1 mg/mL phosphatidylserine, 5 ug/mL diacetyl glyerol, 30 uM ATP, 0.005 uCi/uL 33ATP, 0.25 mg/mL myelin basic protein, serial dilutions of enzastaurin (1-2, 000 nM), and recombinant human PKCbetaII, PKCalpha, PKCepsilon, or PKCgamma enzymes (390, 169, 719, or 128 pM, respectively). Reactions are started by addition of the enzyme and incubated at room temperature for 60 minutes. They are then quenched with 10% H3PO4, transferred to multiscreen anionic phosphocellulose 96-well filter plates, incubated for 30 to 90 minutes, filtered and washed with 4 volumes of 0.5% H3PO4 on a vacuum manifold. Scintillation cocktail is added and plates are read on a Microbeta scintillation counter. IC50 values are determined by fitting a three-variable logistic equation to the 10-point dose-response data using ActivityBase 4.0.

515, 61

DMSO 30 mg/mL, Water <1 mg/mL, Ethanol <1 mg/mL