In today’s global plan to completely eradicate hepatitis C virus (HCV), the essential list
of medications used for HCV treatment are direct-acting antivirals (DAAs), as interferon-sparing
regimens have become the standard-of-care (SOC) treatment. HCV nonstructural protein 5A (NS5A)
inhibitors are a very common component of these regimens. Food and Drug Administration (FDA)-
approved NS5A inhibitors, although very potent, do not have the same potency against all eight
genotypes of HCV. Therefore, this study aims to synthesize NS5A inhibitor analogues with high
potency pan-genotypic activity and high metabolic stability. Starting from an NS5A inhibitor scaffold
previously identified by our research group, we made several modifications. Two series of compounds
were created to test the effect of changing the length and spatial conformation (para-para vs. meta-metapositioned bis-imidazole-proline-carbamate), replacing amide groups in the linker with imidazole
groups, as well as different end-cap compositions and sizes. The frontrunner inhibits genotype 1b
(Con1) replicon, with an EC50 value in the picomolar range, and showed high genotypic coverage
with nanomolar range EC50 values against four more genotypes. This together with its high metabolic
stability (t1
⁄2 > 120 min) makes it a potential preclinical candidate
