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Proteolysis Targeting Chimeras With Reduced Off-targets

ABSTRACT: Proteolysis Targeting Chimeras (PROTACs) are molecules that induce proximity between target proteins and E3 ligases triggering target protein degradation. Pomalidomide, a widely used E3 ligase recruiter in PROTACs, can independently degrade other proteins, including zinc-finger (ZF) proteins, with vital roles in health and disease. This off-target degradation hampers the therapeutic applicability of pomalidomide-based PROTACs, requiring development of PROTAC design rules that minimize off-target degradation. Here we developed a highthroughput platform that interrogates off-target degradation and found that reported pomalidomide-based PROTACs induce degradation of several ZF proteins. We generated a library of pomalidomide analogs to understand how functionalising different positions of the phthalimide ring, hydrogen bonding, steric and hydrophobic effects impact ZF protein degradation. Modifications of appropriate size on the C5 position reduced off-target ZF degradation, which we validated through target engagement and proteomics studies. By applying these design principles, we developed ALK oncoprotein-targeting PROTACs with enhanced potency and minimal offtarget degradation.

INSTRUMENT(S): timsTOF Pro 2

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Eric Fischer

LAB HEAD: Eric Fischer

PROVIDER: PXD046264 | Pride | 2023-12-20

REPOSITORIES: Pride

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			Action	DRS
	esf4_1405_Slot1-1_1_2221.d.zip	Other
	esf4_1406_Slot1-2_1_2222.d.zip	Other
	esf4_1407_Slot1-3_1_2223.d.zip	Other
	esf4_1408_Slot1-4_1_2224.d.zip	Other
	esf4_1409_Slot1-5_1_2226.d.zip	Other

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