High-Throughput Miniaturized Synthesis of PROTAC-Like Molecules.

The development of miniaturized high-throughput in situ screening platforms capable of handling the entire process of drug synthesis to final screening is essential for advancing drug discovery in the future. In this study, an approach based on combinatorial solid-phase synthesis, enabling the efficient synthesis of libraries of proteolysis targeting chimeras (PROTACs) in an array format is presented. This on-chip platform allows direct biological screening without the need for transfer steps.  UV-induced release of target molecules into individual droplets facilitates further on-chip experimentation. Utilizing a mitogen-activated protein kinase kinases (MEK1/2) degrader as a template, a series of 132 novel PROTAC-like molecules is synthesized using solid-phase Ugi reaction. These compounds are further characterized using various methods, including matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) imaging, while consuming only a few milligrams of starting materials in total. Furthermore, the feasibility of culturing cancer cells on the modified spots and quantifying the effect of MEK suppression is demonstrated. The miniaturized synthesis platform lays a foundation for high-throughput in situ biological screening of potent PROTACs for potential anticancer activity and offers the potential for accelerating the drug discovery process by integrating miniaturized synthesis and biological steps on the same array.

Palladium-Catalyzed Combinatorial Synthesis of Biphenyls on Droplet Microarrays at Nanoliter Scale.

The rising costs of pharmaceutical research are currently limiting the productivity of drug discovery and development, but can potentially be diminished via miniaturization of the synthesis and screening of new compounds. As droplet microarrays already present themselves as a versatile tool for highly miniaturized biological screening of various targets, their use for chemical synthesis is still limited. In this study, the influential palladium-catalyzed Suzuki-Miyaura reaction is successfully implemented at the nanoliter scale on droplet microarrays for the synthesis of an 800-compound library of biphenyls. Each reaction is carried out in individual 150 nL droplets. Remarkably, the synthesis of these 800 compounds requires a minimal amount of reagents, totaling 80 µmol, and a solvent volume of 400 µL. Furthermore, the cleavage kinetics and purity of the obtained biphenylic compounds are investigated. Via the solid-phase synthesis approach, the compounds could be purified from excess reactants and catalyst prior to the analysis and a UV-cleavable linker allows for fast and additive-free cleavage of each compound into the individual 100 nL droplet. This novel approach expands the toolbox of the droplet microarray for miniaturized high-throughput chemical synthesis and paves the way for future synthesis and screening of chemical compounds in a single platform.