Parallel Screening of Wild-Type and Drug-Resistant Targets for Anti-Resistance Neuraminidase Inhibitors

Abstract

Infection with influenza virus is a major public health problem, causing serious illness and death each year. Emergence of drug-resistant influenza virus strains limits the effectiveness of drug treatment. Importantly, a dual H275Y/I223R mutation detected in the pandemic influenza A 2009 virus strain results in multidrug resistance to current neuraminidase (NA) drugs. Therefore, discovery of new agents for treating multiple drug-resistant (MDR) influenza virus infections is important. Here, we propose a parallel screening strategy that simultaneously screens wild-type (WT) and MDR NAs, and identifies inhibitors matching the subsite characteristics of both NA-binding sites. These may maintain their potency when drug-resistant mutations arise. Initially, we analyzed the subsite of the dual H275Y/I223R NA mutant. Analysis of the site-moiety maps of NA protein structures show that the mutant subsite has a relatively small volume and is highly polar compared with the WT subsite. Moreover, the mutant subsite has a high preference for forming hydrogen-bonding interactions with polar moieties. These changes may drive multidrug resistance. Using this strategy, we identified a new inhibitor, Remazol Brilliant Blue R (RB19, an anthraquinone dye), which inhibited WT NA and MDR NA with IC50 values of 3.4 and 4.5 µM, respectively. RB19 comprises a rigid core scaffold and a flexible chain with a large polar moiety. The former interacts with highly conserved residues, decreasing the probability of resistance. The latter forms van der Waals contacts with the WT subsite and yields hydrogen bonds with the mutant subsite by switching the orientation of its flexible side chain. Both scaffolds of RB19 are good starting points for lead optimization. The results reveal a parallel screening strategy for identifying resistance mechanisms and discovering anti-resistance neuraminidase inhibitors. We believe that this strategy may be applied to other diseases with high mutation rates, such as cancer and human immunodeficiency virus type 1.

Five Novel Non-Sialic Acid-Like Scaffolds Inhibit In Vitro H1N1 and H5N2 Neuraminidase Activity of Influenza a Virus

“…Inspecting the final docked poses helped identify typical noncovalent interaction patterns such as hydrogen bonds, ionic or saline bridges, van der Waals or hydrophobic interactions, all of which are commonly seen in reversible drug-receptor complexes. Not all observed drug-interacting residues at the active site were also reported in the literature [ 25 , 26 , 27 , 28 , 29 ]. This finding constitutes a key argument in favor of the five scaffolds in view of drug resistance by mutated residues.…”

“…Precisely, the active site of NA is extremely polar. To facilitate the inspection of the inhibitor binding modes, the active site was divided into five subsites (S1-S5) [ 27 , 32 , 33 ] ( Figure 3 ). S1 is composed of three basic amino acids (R118, R292, and R371), whereas S2 is formed by three acidic amino acids (E119, D151, and E227) in addition to W178.…”

Identification of neuraminidase inhibitors against dual H274Y/I222R mutant strains

“…5A ). However, the surface view of the dual mutant structure shows a change in volume and polarity within the binding site due to the H274Y/I222R residue mutation 51 . The I222R mutation has been previously shown to reduce the volume of the binding site 51 .…”

Development of 1‐Amino‐4‐(phenylamino)anthraquinone‐2‐sulfonate Sodium Derivatives as a New Class of Inhibitors of RANKL‐Induced Osteoclastogenesis

“…Dye ligands are considered important alternatives to natural counterparts for binding different types of protein structures because these ligands can interact with active sites of many proteins by mimicking the structures of the substrates, cofactors, or binding agents for those proteins . Bioactive anthraquinone derivatives (Fig.…”

“…Bioactive anthraquinone derivatives (Fig. ) are some of the most widely used reactive dyes in drug discovery that were found to have a diverse range of biological activities such as anti‐inflammatory , anti‐tumor , and anti‐viral activities . Among these anthraquinone dye ligands, it is noteworthy to mention that anilinoanthraquinone derivatives were found to act as antagonists of P2 purinergic receptors , inhibitors of nucleotidases , and activators of large conductance Ca 2+ ‐activated K + channels .…”