Target-induced formation of neuraminidase inhibitors from in vitro virtual combinatorial libraries

Abstract

Neuraminidase, a key enzyme responsible for influenza virus propagation, has been used as a template for selective synthesis of small subsets of its own inhibitors from theoretically highly diverse dynamic combinatorial libraries. We show that the library building blocks, aldehydes and amines, form significant amounts of the library components resulting from their coupling by reductive amination only in the presence of the enzyme. The target amplifies the best hits at least 120-fold. The dynamic libraries synthesized and screened in such an in vitro virtual mode form the components that possess high inhibitory activity, as confirmed by enzyme assays with independently synthesized individual compounds.

Catalytic Self‐Screening of Cholinesterase Substrates from a Dynamic Combinatorial Thioester Library

“…[1][2][3][4][5][6] The technique relies on reversible interconnections between building elements, assembled into discrete entities of interacting species, which constitute libraries of dynamically interchanging species. Different formats of the technique have been developed, including the adaptive approach, [7][8][9][10][11] the pre-equilibrated approach, [12,13] the iterative approach, [14] and the deletion approach, [15] all of which address specific challenges. The concept has also been demonstrated for a range of applications: from model systems to biological applications.…”

“…Until now, mainly imines, [7,10,16] acyl hydrazones, [12,17,18] and disulfides [9,13,19,20] have been used for DCLs since these formats have proven to be the most efficient in the systems studied. This is especially the case when biological systems are targeted, since these require reactions that are stable under mild conditions in the aqueous phase.…”

Kinetic Template‐Guided Tethering of Fragments

“…[47] They used the reversible reaction between aldehydes and a core amine scaffold 1, to probe a hydrophobic pocket on influenza virus A neuraminidase. Amine 1 is based on the commercial neuraminidase inhibitor Tamiflu 2, which binds adjacent to the hydrophobic pocket of interest, Figure 2.…”

A Fragment‐Based Approach to Probing Adenosine Recognition Sites by Using Dynamic Combinatorial Chemistry

“…[9,10] The use of proteins as templates in DCLs for ligand identification has been reported; however, few examples have shown the applicability of this method to assembling noncovalent binding fragments. [11][12][13][14] Recently, LiØnard et al reported the use of DCC coupled with mass spectrometry to identify inhibitors of a metallo-b-lactamase, relying upon the formation of a stable complex between a thiol and two functional zinc ions. [15] Cancilla et al have modified previous "extended tethering" procedures to find inhibitors of Aurora A kinase, thus removing the necessity for a reactive "warhead" to modify the protein, but still requiring the presence of a wild-type or introduced cysteine residue.…”