Novel antiviral agents targeting early stage CMV replication - Summary: Human cytomegalovirus (CMV) is a β-herpes virus with high seroprevalence rates of 60-90% within the population that can spread through bodily fluids, organ transplants, and from mother to fetus via the placenta. Virus proliferation significantly increases the morbidity and mortality of immunocompromised individuals, such as newborns, organ transplant recipients, AIDS patients, and the elderly. Approximately 30,000 solid organ transplant and 23,000 bone marrow transplants operations are performed in the U.S. every year. CMV is the leading cause of birth defects, affecting ~1% of newborns giving rise to ~30,000 new cases of CMV infection reported annually in the US. While several drugs have been approved by the FDA for the treatment of CMV infections, including ganciclovir, foscarnet, letermovir, and recently maribavir, these compounds were found to exhibit high frequencies of drug resistance and severe side effects, including bone marrow toxicity, gastrointestinal disruption, and nephrotoxicity. Given the large number of patient populations at risk for CMV- associated diseases and the estimated cost to treat CMV in the US ($4.4 billion/year by the National Academy of Sciences), novel therapeutic strategies to treat CMV-associated diseases is needed. The development of novel therapeutics that target different steps of the viral life cycle to limit virus propagation and spread would provide therapeutic for treating CMV-related diseases. We developed a high-content screening assay using a CMV AD169 reporter virus to screen >112,000 compounds with Microbiotix to identify inhibitors that block the early stage of infection. One compound, MBXC-4302, is a N-arylpyrimidinamine (NAPA) that exhibited potent anti-CMV infection activity (IC50 ~3 µM), limited cytotoxicity (CC50 >100µM), favorable in vitro ADME properties, and a responsive structure activity relationship (SAR). The preliminary SAR analysis identified the analogs with an improved selective index. Our general hypothesis is that the NAPA compounds represent novel CMV entry inhibitors which can be developed into effective novel CMV therapeutics to prevent virus proliferation and spread. To evaluate our hypothesis, we plan to complete the following Aims: 1) Characterize the mechanism of action and target of NAPA compounds; 2) Optimize the NAPA series through SAR-driven analog generation; 3) Evaluate NAPA compounds to limit virus proliferation using in vitro models; and 4) Evaluate pharmacokinetics, tolerability, and efficacy of prioritized NAPA analogs in proliferation studies in vivo We plan to develop 1-2 lead NAPA analogs as effective CMV therapeutics that inhibit CMV dissemination as monotherapy or combination therapy with FDA approved CMV drugs to prevent virus-associated diseases by limiting viral load.
