Driving factors in amiloride recognition of HIV RNA targets

Abstract

RNA profiling reveals key contributors to affinity and selectivity in amiloride : RNA interactions, including a predictive model for ESSV recognition.

IRES-targeting small molecule inhibits enterovirus 71 replication via allosteric stabilization of a ternary complex

“…By comparison, all six SMs caused significant CSPs to adenosines adjacent to (A133 and A139) and located within (A134 and A136) the bulge. Thus, these data clearly localize the SLII-binding sites for the SMs to the surface formed by the bulge, in line with previous preferences for amiloride derivatives 22,23 (Fig. 1b).…”

“…1 ), which facilitated the development of a high-throughput FID protocol to screen SMs against low concentrations of SLII ( Z score 0.49, Supplementary Note 2 ). The SM library largely contained derivatives of amiloride, previously identified as a tunable RNA-binding scaffold 22 , 23 (Supplementary Fig. 2 ), along with other known RNA-binding SMs.…”

“…Concentration of samples was determined using the RNA theoretical molar extinction coefficient, and NMR samples ranged from 0.05 to 0. 22,23 . 4-Trifluoromethyl derivative DMA-197 was synthesized via the previously reported two step procedure using Sonogashira coupling reaction followed by guanidinylation 22 .…”

Small Molecule Targeting IRES Domain Inhibits Enterovirus 71 Replication via an Allosteric Mechanism that Stabilizes a Ternary Complex

“…We observed tight binding of the indicator peptide to SLII (24.5 ± 4.7 nM - Figure S1), which facilitated the development of a highthroughput FID protocol to screen small molecules against low concentrations of SLII (Z' score 0.49, see SI). The small molecule library largely contained derivatives of amiloride, previously identified as a tunable RNA-binding scaffold 21,22 (Figure S2), along with other known RNA binding small molecules. The screening assay was performed at two different small molecule concentrations to identify both strong and weak binding derivatives, and "hits" were defined as causing fluorescent changes greater than 25%.…”

“…Synthesis of amiloride derivatives DMA-001, DMA-019, DMA-096, DMA-097, DMA-101, DMA-132, DMA-135, DMA-155, DMA-165, DMA-169, DMA-178, and DMA-185 have been previously reported 21,22 . 4-Trifluoromethyl derivative DMA-197 was synthesized via the previously reported 2 step procedure using Sonogashira coupling reaction followed by guanidinylation 21 .…”