COVID-19 Genetics and Drug Development

In December of 2019, the World Health Organization (WHO) was notified by Chinese officials of a pneumonia of unknown etiology that had affected nearly 44 individuals. On January 7, 2020, Chinese researchers isolated a novel coronavirus now known as SARS-CoV-2 that causes the disease COVID-19. Using our knowledge and expertise in clinical pharmacology and genomics, we investigate the genetic differences affecting the progress of SARS-CoV-2 infection and target the development of novel assays or therapeutics for COVID-19. We focus our efforts on the following three pilot projects. (1) Development and Validation of a Simple, Selective and Rapid LC-MS/MS Assay for the Quantification of Remdesivir in Humans with SARS-CoV-2: Remdesivir, formerly GS-5734, has recently become the first antiviral drug approved by the U.S. Food and Drug Administration (FDA) to treat COVID-19, the disease caused by SARS-CoV-2. Therapeutic dosing and pharmacokinetic studies require a simple, sensitive, and selective validated assay to quantify drug concentrations in clinical samples. Therefore, we developed a rapid and sensitive LC-MS/MS assay for the quantification of remdesivir in human plasma with its deuterium-labeled analog, remdesivir-2H5, as the internal standard. Chromatographic separation was achieved on a Phenomenex Synergi HPLC Fusion-RP column by gradient elution. Excellent accuracy and precision (5.2% within-run variations and. 9.8% between-run variations) were obtained over the range of 0.5-5000 ng/mL. The assay met the FDA Bioanalytical Guidelines for selectivity and specificity, and low inter-matrix lot variability (2.7%) was observed for extraction efficiency (77%) and matrix effect (123%) studies. Further, stability tests showed that the analyte does not degrade under working conditions, nor during freezing and thawing processes. (2) Development of TMPRSS2 Targeted Therapies for COVID-19: The angiotensin I converting enzyme 2 (ACE2) and the transmembrane protease, serine 2 (TMPRSS2) have been implicated in influenza and SARS-CoV infection as well as for SARS-CoV-2 in mediating viral entry into the host cell. We have identified potential compounds of interest that target TMPRSS2 or other mechanisms that mediate viral entry and have initiated preclinical studies with collaborators in Japan and at NCATS to screen these compounds in live SARS-CoV-2 virus assays. The overall goal is to identify novel potential TMPRSS2 inhibitor as a part of a rational therapeutic combination for treatment of patients with COVID-19. (3) TMPRSS2 Polymorphisms on Sex Specific Outcomes in COVID-19: The COVID-19 data so far have shown that the mortality rate for men is higher than women. Initial reports and those recently from the scientific community attribute the discrepancy to smoking habits, sex-based disparities in the immune response or other comorbidities. We proposed that the observed sex differences in clinical outcomes in patients may involve genomic differences in genes that drive SARS-CoV-2 infections. Based on data from the literature, we propose that TMPRSS2, an androgen-regulated protein that mediates entry of SARS-CoV-2 into the host cell, may contribute to the severity of COVID-19 outcomes. We have initiated a collaboration with Columbia University to have access to their clinical specimens collected from COVID-19 patients. We propose to genotype patients who are positive for COVID-19 to evaluate known single nucleotide polymorphisms and correlate to disease susceptibility and/or disease outcomes. Underlying genetic polymorphisms may contribute to the observed sex differences noted in the severity of clinical outcomes in patients with COVID-19. Furthermore, understanding the genetic differences that lead to changes in clinical outcomes, especially clear sexual dimorphisms, can help inform patterns of susceptibility (if any) and future targeted drug therapy research and development, and more importantly, disease management. The protocol has already been written and submitted for IRB review at Columbia University. The expected outcome for this project is that discovery of polymorphisms in TMPRSS2 could lead to TMPRSS2 becoming an actionable biomarker, stratifying patients by severity and informing healthcare providers the patients that will require the most care. (4) The Biospecimen Processing Core (BPC) is currently involved in processing research bloods for a COVID-19 clinical protocol. Progress to data: For project 1, the remedesivir PK bioanalytical assay has been developed, validated, and published. For project 2, at this time, no lead compound has been identified from the drug development screen efforts. For project 3, we have not received COVID-19 clinical samples from Columbia University. For project 4, the BPC is currently processing research samples for a COVID-19 clinical protocol.