Therapeutic combinations comprising a raf inhibitor and a erk inhibitor
A technology of inhibitors, kinase inhibitors, used in c-RAF inhibitors, ERK inhibitors, pharmaceutical compositions containing such combinations, therapeutic combinations for the treatment of cancer, in the field of cancer treatment, to address unmet medical needs, etc. question
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[0105] Example 1 : N-(3-(2-(2-hydroxyethoxy)-6-morpholinopyridin-4-yl)-4-methylphenyl)-2-(trifluoromethyl)isonicotinamide
[0106] Compound A (Compound A) is a morpholine-substituted biaryl compound having the structure
[0107]
example 1
[0110] In vitro Raf activity assay
[0111] Both the RAF enzyme and the catalytically inactive MEK1 protein substrate were prepared in-house using conventional methods. The CRAF cDNA as a full-length protein (with Y340E and Y341E activating mutations) was subcloned into a baculovirus expression vector for Sf9 insect cell expression. The h14-3-3ζ cDNA was subcloned into a baculovirus expression vector for expression in SF9 insect cells. Sf9 cells co-expressing the two proteins were lysed and subjected to immobilized nickel chromatography, eluted with imidazole. A second column (StrepII binding column) was used and eluted with desthiobiotin. Protein tags are removed using Prescission enzyme and the protein is further purified using a flow-through step to remove tags.
[0112] C-Raf FL refers to full length C-Raf protein.
[0113] Full-length MEK1 with an inactivating K97R ATP-binding site mutation was used as a RAF substrate. Use the N-terminal (his) of MEK1 cDNA 6 Tags we...
example 1
[0136] Compound A exhibited activity in a number of human cancer cell lines expressing mutations in the MAPK pathway, as shown in the table below. Note that activity is particularly strong for cell lines with at least one mutation in BRAF or RAS.
[0137] Table 1. Effect of Compound A on the proliferation of a panel of human cancer cell lines.
[0138]
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