Journal: RSC Chemical Biology

Article Title: Development of PROTACs for targeted degradation of oncogenic TRK fusions

doi: 10.1039/d6cb00008h

Figure Lengend Snippet: Design and structures of entrectinib-derived PROTACs. (A) The co-crystal structure of entrectinib with TRKA kinase domain (PDB ID: 5KVT), in which the N -methyl piperazine is solvent accessible, as indicated by the red arrow. (B) The proposed modification on the piperazine ring for the linker introduction. (C and D) Chemical structures of the indicated compounds.

Article Snippet: Entrectinib (Selleck Chemicals, #S7998), GNF-8625 (MedChemExpress, #HY-131706A), CG-428 (Tocris, #7425), carfilzomib (MedChemExpress, #HY-10455), MLN4924 (Calbiochem, #5054770001), lenalidomide (Millipore Sigma, #901558), bafilomycin A1 (Cambridge Bioscience, #b0025) and pomalidomide (MedChemExpress, #HY-10984) were purchased from the indicated sources.

Techniques: Derivative Assay, Solvent, Modification

Journal: RSC Chemical Biology

Article Title: Development of PROTACs for targeted degradation of oncogenic TRK fusions

doi: 10.1039/d6cb00008h

Figure Lengend Snippet: JWJ-01-378 induces potent degradation of TPM3-TRKA. (A) Immunoblot analysis of KM12 cells treated with DMSO, CG-428, JWJ-01-375, JWJ-01-377, JWJ-01-378, and entrectinib at the indicated doses for 2 hours. (B) Immunoblot analysis of KM12 cells treated with DMSO, CG-428, JWJ-01-375, JWJ-01-377, JWJ-01-378, and entrectinib at the indicated doses for 6 hours. (C) Immunoblot analysis of KM12 cells treated with DMSO, JWJ-01-378 and JWJ-01-392 at the indicated doses for 2 hours. (D) Immunoblot analysis of KM12 cells treated with DMSO, CG-428, JWJ-01-375, JWJ-01-377, JWJ-01-378, and JWJ-01-392 at the indicated doses for 6 hours. (E) Immunoblot analysis of KM12 cells treated with DMSO, 10 nM JWJ-01-378, 10 nM JWJ-01-392 and 10 nM entrectinib for the indicated time-course. (F) Immunoblot analysis of KM12 cells treated with DMSO, 10 nM CG-428, 10 nM JWJ-01-378, and 10 nM JWJ-01-392 for the indicated time-course. (G) Immunoblot analysis of KM12 cells pretreated with 0.5 µM of carfilzomib (Carf), 1 µM MLN4924 (MLN), 0.1 µM entrectinib, 10 µM lenalidomide (Len), 1 µM JWJ-01-392, or 0.1 µM bafilomycin A1 (BafA1) for 2 hours followed by DMSO or 10 nM JWJ-01-378 treatment for 4 hours. Data in (A–G) are representative of n = 3 independent experiments. (H) Protein abundance after treatment of KM12 cells with 100 nM of JWJ-01-378 for 2 hours compared to DMSO. Volcano plots depict fold change abundance relative to DMSO versus P value derived from a moderated t -test. Fold change values and significance designations derived from a moderated t -test are provided in Dataset S1. Data are from n = 3 biologically independent samples.

Article Snippet: Entrectinib (Selleck Chemicals, #S7998), GNF-8625 (MedChemExpress, #HY-131706A), CG-428 (Tocris, #7425), carfilzomib (MedChemExpress, #HY-10455), MLN4924 (Calbiochem, #5054770001), lenalidomide (Millipore Sigma, #901558), bafilomycin A1 (Cambridge Bioscience, #b0025) and pomalidomide (MedChemExpress, #HY-10984) were purchased from the indicated sources.

Techniques: Western Blot, Quantitative Proteomics, Derivative Assay