Sigma Receptor Ligands Prevent COVID Mortality In Vivo: Implications for Future Therapeutics
Reed L Berkowitz, Andrew P Bluhm, Glenn W Knox, Christopher R Mccurdy, David A Ostrov, Michael H Norris
International Journal of Molecular Sciences, doi:10.3390/ijms242115718
The emergence of lethal coronaviruses follows a periodic pattern which suggests a recurring cycle of outbreaks. It remains uncertain as to when the next lethal coronavirus will emerge, though its eventual emergence appears to be inevitable. New mutations in evolving SARS-CoV-2 variants have provided resistance to current antiviral drugs, monoclonal antibodies, and vaccines, reducing their therapeutic efficacy. This underscores the urgent need to investigate alternative therapeutic approaches. Sigma receptors have been unexpectedly linked to the SARS-CoV-2 life cycle due to the direct antiviral effect of their ligands. Coronavirus-induced cell stress facilitates the formation of an ER-derived complex conducive to its replication. Sigma receptor ligands are believed to prevent the formation of this complex. Repurposing FDA-approved drugs for COVID-19 offers a timely and cost-efficient strategy to find treatments with established safety profiles. Notably, diphenhydramine, a sigma receptor ligand, is thought to counteract the virus by inhibiting the creation of ER-derived replication vesicles. Furthermore, lactoferrin, a well-characterized immunomodulatory protein, has shown antiviral efficacy against SARS-CoV-2 both in laboratory settings and in living organisms. In the present study, we aimed to explore the impact of sigma receptor ligands on SARS-CoV-2-induced mortality in ACE2-transgenic mice. We assessed the effects of an investigational antiviral drug combination comprising a sigma receptor ligand and an immunomodulatory protein. Mice treated with sigma-2 receptor ligands or diphenhydramine and lactoferrin exhibited improved survival rates and rapid rebound in mass following the SARS-CoV-2 challenge compared to mock-treated animals. Clinical translation of these findings may support the discovery of new treatment and research strategies for SARS-CoV-2.
Institutional Review Board Statement: The studies and SOPs were approved by the University of Florida Institutional Biosafety Committee under protocol BIO5594. They were carried out in the University of Florida Environmental Health and Safety inspected and approved BSL3 and ABSL3 facilities. Ethical approval for the animal studies and procedures in this work were approved by the University of Florida IACUC under protocol #202111322. Humane endpoints and euthanasia guidelines according to the American Veterinary Medical Association were adhered to throughout the study.
Informed Consent
References
Campione, Lanna, Cosio, Rosa, Conte et al., Lactoferrin as Antiviral Treatment in COVID-19 Management: Preliminary Evidence, Int. J. Environ. Res. Public. Health,
doi:10.3390/ijerph182010985Cappanera, Palumbo, Kwan, Priante, Martella et al., When Does the Cytokine Storm Begin in COVID-19 Patients? A Quick Score to Recognize It, J. Clin. Med,
doi:10.3390/jcm10020297Charness, Gupta, Stack, Strymish, Adams et al., Rebound of SARS-CoV-2 Infection after Nirmatrelvir-Ritonavir Treatment, N. Engl. J. Med,
doi:10.1056/NEJMc2206449Cirino, Eans, Medina, Wilson, Mottinelli et al., Characterization of Sigma 1 Receptor Antagonist CM-304 and Its Analog, AZ-66: Novel Therapeutics Against Allodynia and Induced Pain, Front. Pharmacol,
doi:10.3389/fphar.2019.00678Crespo, Calleja, Fernández, Sacristan, Ruiz-Antorán et al., Real-World Effectiveness and Safety of Oral Combination Antiviral Therapy for Hepatitis C Virus Genotype 4 Infection, Clin. Gastroenterol. Hepatol,
doi:10.1016/j.cgh.2017.02.020Csr Mers Outbreaks, None
Fung, Huang, Liu, Coronavirus-Induced ER Stress Response and Its Involvement in Regulation of Coronavirus-Host Interactions, Virus Res,
doi:10.1016/j.virusres.2014.09.016Gordon, Jang, Bouhaddou, Xu, Obernier et al., A SARS-CoV-2 Protein Interaction Map Reveals Targets for Drug Repurposing, Nature,
doi:10.1038/s41586-020-2286-9Grodzki, Bluhm, Schaefer, Tagmount, Russo et al., Genome-Scale CRISPR Screens Identify Host Factors That Promote Human Coronavirus Infection, Genome Med,
doi:10.1186/s13073-022-01013-1Hu, Lewandowski, Tan, Zhang, Morgan et al., Naturally Occurring Mutations of SARS-CoV-2 Main Protease Confer Drug Resistance to Nirmatrelvir
Hu, Meng, Zhang, Xiang, Wang, The in Vitro Antiviral Activity of Lactoferrin against Common Human Coronaviruses and SARS-CoV-2 Is Mediated by Targeting the Heparan Sulfate Co-Receptor, Emerg. Microbes Infect,
doi:10.1080/22221751.2021.1888660Intagliata, Sharma, King, Mesangeau, Seminerio et al., Discovery of a Highly Selective Sigma-2 Receptor Ligand, 1-(4-(6,7-Dimethoxy-3,4-Dihydroisoquinolin-2(1H)-Yl)Butyl)-3-Methyl-1H-Benzo[d]Imidazol-2(3H)-One (CM398), with Drug-Like Properties and Antinociceptive Effects In Vivo, AAPS J,
doi:10.1208/s12248-020-00472-xJames, Shen, Zavaleta, Nielsen, Mesangeau et al., New Positron Emission Tomography (PET) Radioligand for Imaging σ-1 Receptors in Living Subjects, J. Med. Chem,
doi:10.1021/jm300371cJochmans, Liu, Donckers, Stoycheva, Boland et al., The Substitutions L50F, E166A and L167F in SARS-CoV-2 3CLpro Are Selected by a Protease Inhibitor In Vitro and Confer Resistance to Nirmatrelvir
Kabinger, Stiller, Schmitzová, Dienemann, Kokic et al., Mechanism of Molnupiravir-Induced SARS-CoV-2 Mutagenesis, Nat. Struct. Mol. Biol,
doi:10.1038/s41594-021-00651-0Kaufmann, Dorhoi, Hotchkiss, Bartenschlager, Host-Directed Therapies for Bacterial and Viral Infections, Nat. Rev. Drug Discov,
doi:10.1038/nrd.2017.162Kozu, Iinuma, Ohashi, Saito, Akasu et al., Effect of Orally Administered Bovine Lactoferrin on the Growth of Adenomatous Colorectal Polyps in a Randomized, Placebo-Controlled Clinical Trial, Cancer Prev. Res,
doi:10.1158/1940-6207.CAPR-08-0208Lednicky, Waltzek, Mcgeehan, Loeb, Hamilton et al., Isolation and Genetic Characterization of Human Coronavirus NL63 in Primary Human Renal Proximal Tubular Epithelial Cells Obtained from a Commercial Supplier, and Confirmation of Its Replication in Two Different Types of Human Primary Kidney Cells, Virol. J,
doi:10.1186/1743-422X-10-213Matino, Tavella, Rizzi, Avanzi, Azzolina et al., Effect of Lactoferrin on Clinical Outcomes of Hospitalized Patients with COVID-19: The LAC Randomized Clinical Trial, Nutrients,
doi:10.3390/nu15051285Matthew, Leidner, Lockbaum, Henes, Zephyr et al., Drug Design Strategies to Avoid Resistance in Direct-Acting Antivirals and Beyond, Chem. Rev,
doi:10.1021/acs.chemrev.0c00648Merlos, Romero, Zamanillo, Plata-Salamán, Vela, Sigma-1 Receptor and Pain, Handb. Exp. Pharmacol,
doi:10.1007/164_2017_9Mihelc, Baker, Lanman, Coronavirus Infection Induces Progressive Restructuring of the Endoplasmic Reticulum Involving the Formation and Degradation of Double Membrane Vesicles, Virology,
doi:10.1016/j.virol.2020.12.007Mirabelli, Wotring, Zhang, Mccarty, Fursmidt et al., Morphological Cell Profiling of SARS-CoV-2 Infection Identifies Drug Repurposing Candidates for COVID-19, Proc. Natl. Acad. Sci,
doi:10.1073/pnas.2105815118Moore, Anderson, Groom, Haridas, Baker, Three-Dimensional Structure of Diferric Bovine Lactoferrin at 2.8 Å Resolution, J. Mol. Biol,
doi:10.1006/jmbi.1997.1386Moreno, Perno, Mallon, Behrens, Corbeau et al., Two-Drug vs. Three-Drug Combinations for HIV-1: Do We Have Enough Data to Make the Switch?, HIV Med,
doi:10.1111/hiv.12716Naguib, Li, Ling, Grace, Nguyen-Viet et al., Live and Wet Markets: Food Access versus the Risk of Disease Emergence, Trends Microbiol,
doi:10.1016/j.tim.2021.02.007Navarro, Paredes, Tucto, Medina, Angles-Yanqui et al., Bovine Lactoferrin for the Prevention of COVID-19 Infection in Health Care Personnel: A Double-Blinded Randomized Clinical Trial (LF-COVID), BioMetals,
doi:10.1007/s10534-022-00477-3Oladunni, Park, Pino, Gonzalez, Akhter et al., Lethality of SARS-CoV-2 Infection in K18 Human Angiotensin-Converting Enzyme 2 Transgenic Mice, Nat. Commun,
doi:10.1038/s41467-020-19891-7Ostrov, Bluhm, Li, Khan, Rohamare et al., Highly Specific Sigma Receptor Ligands Exhibit Anti-Viral Properties in SARS-CoV-2 Infected Cells, Pathogens,
doi:10.3390/pathogens10111514Piacentini, Centi, Miotto, Milanetti, Di Rienzo et al., Lactoferrin Inhibition of the Complex Formation between ACE2 Receptor and SARS CoV-2 Recognition Binding Domain, Int. J. Mol. Sci,
doi:10.3390/ijms23105436Presti, Manti, Parisi, Papale, Barbagallo et al., Lactoferrin: Cytokine Modulation and Application in Clinical Practice, J. Clin. Med,
doi:10.3390/jcm10235482Rathnasinghe, Strohmeier, Amanat, Gillespie, Krammer et al., Comparison of Transgenic and Adenovirus hACE2 Mouse Models for SARS-CoV-2 Infection, Emerg. Microbes Infect,
doi:10.1080/22221751.2020.1838955Reznikov, Norris, Vashisht, Bluhm, Li et al., Identification of Antiviral Antihistamines for COVID-19 Repurposing, Biochem. Biophys. Res. Commun,
doi:10.1016/j.bbrc.2020.11.095Ricciardi, Guarino, Giaquinto, Polishchuk, Santoro et al., The Role of NSP6 in the Biogenesis of the SARS-CoV-2 Replication Organelle, Nature,
doi:10.1038/s41586-022-04835-6Rosa, Tripepi, Naldi, Aimati, Santangeli et al., Ambulatory COVID-19 Patients Treated with Lactoferrin as a Supplementary Antiviral Agent: A Preliminary Study, J. Clin. Med,
doi:10.3390/jcm10184276Sambo, Lebowitz, Khoshbouei, The Sigma-1 Receptor as a Regulator of Dopamine Neurotransmission: A Potential Therapeutic Target for Methamphetamine Addiction, Pharmacol. Ther,
doi:10.1016/j.pharmthera.2018.01.009Sanderson, Hisner, Donovan-Banfield, Hartman, Løchen et al., A Molnupiravir-Associated Mutational Signature in Global SARS-CoV-2 Genomes, Nature,
doi:10.1038/s41586-023-06649-6Schafer, Cheng, Xiong, Soloveva, Retterer et al., Repurposing Potential of 1st Generation H1-Specific Antihistamines as Anti-Filovirus Therapeutics, Antiviral Res,
doi:10.1016/j.antiviral.2018.07.003Seminerio, Robson, Abdelazeem, Mesangeau, Jamalapuram et al., Synthesis and Pharmacological Characterization of a Novel Sigma Receptor Ligand with Improved Metabolic Stability and Antagonistic Effects Against Methamphetamine, AAPS J,
doi:10.1208/s12248-011-9311-8Serrano, Kochergina, Albors, Diaz, Oroval et al., Liposomal Lactoferrin as Potential Preventative and Cure for COVID-19, Int. J. Res. Health Sci,
doi:10.5530/ijrhs.8.1.3Sicari, Zabbo, Diphenhydramine, None
Towler, Staker, Prasad, Menon, Tang et al., ACE2 X-Ray Structures Reveal a Large Hinge-Bending Motion Important for Inhibitor Binding and Catalysis, J. Biol. Chem,
doi:10.1074/jbc.M311191200Yan, Zhang, Li, Xia, Guo et al., Structural Basis for the Recognition of SARS-CoV-2 by Full-Length Human ACE2, Science,
doi:10.1126/science.abb2762Yan, Zhang, Zhou, Li, Huang et al., A Web Server for Protein-Protein and Protein-DNA/RNA Docking Based on a Hybrid Strategy, Nucleic Acids Res,
doi:10.1093/nar/gkx407Yang, Wang, Sun, The Roles of Intracellular Chaperone Proteins, Sigma Receptors, in Parkinson's Disease (PD) and Major Depressive Disorder (MDD), Front. Pharmacol,
doi:10.3389/fphar.2019.00528Zheng, Wong, Li, Verma, Ortiz et al., COVID-19 Treatments and Pathogenesis Including Anosmia in K18-hACE2 Mice, Nature,
doi:10.1038/s41586-020-2943-zZhou, Gammeltoft, Ryberg, Pham, Tjørnelund et al., Nirmatrelvir-Resistant SARS-CoV-2 Variants with High Fitness in an Infectious Cell Culture System, Sci. Adv,
doi:10.1126/sciadv.add7197
