Rapid PROTAC Discovery Platform: Nanomole-Scale Array Synthesis and Direct Screening of Reaction Mixtures

Area of Science:

• Medicinal Chemistry
• Drug Discovery
• Chemical Biology

Background:

• Designing effective proteolysis targeting chimeras (PROTACs) requires precise control over length, shape, and linker attachment points.
• The synthetic complexity and limitations in computational modeling of PROTACs necessitate empirical exploration of structure-activity relationships (SAR), demanding significant time and resources.

Purpose of the Study:

• To develop a rapid, economical, and sustainable methodology for PROTAC synthesis and hit finding.
• To accelerate the exploration of PROTAC SAR by enabling direct screening of reaction mixtures.

Main Methods:

• Developed capabilities for parallel synthesis and purification of PROTACs using preformed E3-ligand-linker intermediates.
• Established a rapid, nanomole-scale PROTAC synthesis methodology utilizing amide coupling.
• Enabled direct screening of nonpurified reaction mixtures in cell-based degradation assays, logD, and EPSA measurements.

Main Results:

• Significantly expanded and accelerated PROTAC SAR exploration, reducing the timeline from several weeks to 5 days.
• Avoided laborious and solvent-intensive purification steps, making the methodology more economical and sustainable.
• Facilitated rapid hit finding for PROTAC development.

Conclusions:

• The developed methodology offers a highly efficient and streamlined approach to PROTAC synthesis and SAR exploration.
• This economical and sustainable method accelerates the discovery of novel PROTACs for therapeutic applications.
• The direct screening of nonpurified mixtures represents a significant advancement in PROTAC hit identification strategies.