# Pharmacophore‐Based Identification and Molecular Characterization of Potent Neprilysin Inhibitors: Biochemical and Therapeutic Implications for Cardiovascular Diseases

## TL;DR

Researchers identified two natural compounds that effectively inhibit neprilysin, a target for heart failure treatments, using computational and biochemical methods.

## Contribution

A pharmacophore-based virtual screening approach combined with MD simulations identified novel, non-toxic neprilysin inhibitors.

## Key findings

- Pentagalloylglucose and tannic acid inhibit neprilysin with IC50 values of 17.2 ± 1.5 μM and 10.9 ± 0.7 μM.
- Molecular dynamics simulations revealed stable interactions with the catalytic Zn2+ site.
- The compounds showed negligible cytotoxicity in HEK293T cells.

## Abstract

Neprilysin (NEP), a zinc‐dependent metalloprotease involved in the degradation of bioactive peptides, represents a validated target for heart failure therapeutics. In this study, a pharmacophore‐based virtual screening approach combined with biochemical and biophysical assays, alongside molecular dynamics (MD) simulations, was employed to identify novel NEP inhibitors. The pharmacophore model Phar‐A3D2R1 successfully identified pentagalloylglucose (PGG) and tannic acid as potent inhibitors, with IC50 values of 17.2 ± 1.5 μM and 10.9 ± 0.7 μM, respectively. Local surface plasmon resonance (LSPR) assays confirmed strong binding affinities (KD = 6.2 ± 0.4 μM for PGG and 5.9 ± 0.5 μM for tannic acid). MD simulations revealed stable ligand–enzyme interactions mediated by hydrogen bonding, hydrophobic contacts, electrostatic interactions, and coordination with the catalytic Zn2+ ion. Cytotoxicity assessment in HEK293T cells indicated negligible toxicity. These results validate PGG and tannic acid as promising lead compounds for NEP inhibition and provide a basis for further structure‐based optimization toward cardiovascular therapeutics.

Pharmacophore‐based virtual screening and molecular dynamics identified pentagalloylglucose and tannic acid as potent, non‐toxic neprilysin inhibitors. These natural compounds demonstrated strong binding affinity and stable interactions with the catalytic Zn2+ site, offering promising leads for cardiovascular drug development.