Journal: Journal of Medicinal Chemistry

Article Title: Second-Generation AURKA-Targeting PROTACs: Structural Optimization toward in Vivo Degradation in Neuroblastoma

doi: 10.1021/acs.jmedchem.5c01271

Figure Lengend Snippet: (A) Chemical structures of selective AURKA inhibitors MK-5108, LY3295668, and TAS-119. (B) Previously developed AURKA degraders SK2188 and SK3277 generated by coupling MK-5108 to thalidomide using PEG-based linkers of varying lengths. Both PROTACs exhibited potent biological activities (DC 50 , D max , GI 50 ) and, in the case of SK2188, a high binding selectivity for AURKA over AURKB ( K i AURKA/AURKB) and confirmed binding with CRBN ( K i CRBN). However, both PROTACs showed low plasma stabilities (plasma t 1/2 ), high turnover rates in mouse liver microsomes (CL int MLM), and in the case of SK2188, a high intravenous clearance (CL IV ) (n.d. = not determined).

Article Snippet: For promising candidates, we more accurately assessed their degradation potency (DC 50 , D max ) (Simple Western) ( Supplementary Figure S2B,C ) and monitored their stability in plasma (plasma t 1/2 ), mouse liver microsomes (CL int MLM), and mouse hepatocytes (CL int MH) using Eurofins Discovery Services.

Techniques: Generated, Binding Assay, Clinical Proteomics

Journal: Journal of Medicinal Chemistry

Article Title: Second-Generation AURKA-Targeting PROTACs: Structural Optimization toward in Vivo Degradation in Neuroblastoma

doi: 10.1021/acs.jmedchem.5c01271

Figure Lengend Snippet: Key compounds developed during the PROTAC structural optimization study and their corresponding in vitro and in vivo parameters. Key compounds are categorized into series based on different optimization strategies: ( series 1 ) linker rigidification, ( series 2 ) exploration of arylaminoglutarimide- and ( series 3 ) dihydrouracil-based CRBN ligands, and ( series 4 ) the use of a new AURKA ligand analogous to LY3295668. Antiproliferative effects were monitored using IncuCyte live-cell imaging in NGP and expressed as GI 50 72h after treatment ( n = 3) (see also Supplementary Figure S1 ). AURKA degradation potency was assessed using Simple Western after 24 h treatment in NGP and expressed as DC 50 and D max ( n = 3) (see also Supplementary Figure S2B ). Plasma half-life ( t 1/2 ) and metabolic stability (CL int ) were measured by Eurofins Discovery services through incubation of the compounds in mouse plasma, mouse liver microsomes (MLM), or mouse hepatocytes (MH). The percentage of the compound remaining over time was measured by LC–MS analysis ( n = 2 replicate samples). Binding to AURKA, AURKB, and CRBN was measured using the KINOMEscan or E3scan profiling services by Eurofins Discovery and expressed as K i ( n = 2 replicate samples) (see also Supplementary Figure S3 ). In vivo systemic clearance (CL IV ) was obtained by a noncompartmental analysis following IV administration at a dose of 15 mg/kg in mice ( n = 3 mice) (see also Supplementary Figure S4 ). Data for all synthesized compounds can be found in Supplementary Tables 1–4 .

Article Snippet: For promising candidates, we more accurately assessed their degradation potency (DC 50 , D max ) (Simple Western) ( Supplementary Figure S2B,C ) and monitored their stability in plasma (plasma t 1/2 ), mouse liver microsomes (CL int MLM), and mouse hepatocytes (CL int MH) using Eurofins Discovery Services.

Techniques: In Vitro, In Vivo, Live Cell Imaging, Simple Western, Clinical Proteomics, Incubation, Liquid Chromatography with Mass Spectroscopy, Binding Assay, Synthesized