A direct-to-biology high-throughput chemistry approach to reactive fragment screening

Abstract

Methods for rapid identification of chemical tools are essential for the validation of emerging targets and to provide medicinal chemistry starting points for the development of new medicines. Here, we...

Reactive fragments targeting carboxylate residues employing direct to biology, high-throughput chemistry

“…Hits afforded higher crosslinking yields than observed previously with diazirine-based fragment screening (PhABits), which facilitates both hit calling and follow-up studies. 14,42 This is consistent with the formation of a longer-lived nitrilimine intermediate upon photoactivation of the ACT, which reacts preferentially with carboxylate protein side chains. By contrast, the diazirine forms the shorter-lived carbene upon photoactivation, which can be rapidly quenched by water, often leading to low crosslinking yields.…”

“…Previously we have reported the screening of diazirine containing fragments for the discovery of ligands for proteins of interest. 14,42 A challenge with this approach can be the low crosslinking yields achieved with carbenes, and it was anticipated that the nitrilimine furnished by the photoactivation of the ACT reactive group may offer improved crosslinking yields.…”

Profiling Sulfur(VI) Fluorides as Reactive Functionalities for Chemical Biology Tools and Expansion of the Ligandable Proteome

“…Chemical probes offer a molecular toolkit for the study of the proteome and validation of potential therapeutic targets . Probes comprising reactive functionalities are particularly powerful for the study of protein targets via the covalent modification of selected amino acids. , Covalent inhibitors and therapeutics are a proven strategy to enhance potency and selectivity and to reduce dosing frequency (Figure a­(i)). − Reactive tools have also been employed broadly in chemical biology, enabling robust protein capture and providing access to a suite of techniques, including chemoproteomic mapping of ligand–protein interactions across the proteome (Figure a­(ii)). , More recently, reactive fragment-based screening platforms have been developed for streamlined and robust detection of hits, both with purified proteins of interest and in proteome-wide screening, which offers a route to expand the liganded proteome (Figure a­(iii)). − …”

High-throughput crystallography for rapid fragment growth from crude arrays by low-cost robotics

“…In recent years, progress has been made in developing low-cost HTS platforms [23] for the synthesis, purification and analysis of compounds. [24][25][26][27] We previously completed a fragment screen against the second bromodomain of the pleckstrin homology domain-interacting protein (PHIP(2)), a multidomain protein involved in various cellular processes including cellular growth and mobility [18] that has been implicated in a number of aggressive cancers including BRAFnegative melanomas, breast and lung cancer. [28,29] Following up on the piperazine hit, F709, from this screen, we implemented a growth array synthesis exercise using a low-cost OpenTrons OT-1 TM robotics-based synthesis platform, which allows for a quick survey of the chemical space around a fragment hit.…”