The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has prompted great interest in novel broad-spectrum antivirals, including perylene-related compounds. In the present study, we performed a structure-activity relationship analysis of a series of perylene derivatives, which comprised a large planar perylene residue, and structurally divergent polar groups connected to the perylene core by a rigid ethynyl or thiophene linker. Most of the tested compounds did not exhibit significant cytotoxicity towards multiple cell types susceptible to SARS-CoV-2 infection, and did not change the expressions of cellular stress-related genes under normal light conditions. These compounds showed nanomolar or sub-micromolar dose-dependent anti-SARS-CoV-2 activity, and also suppressed the in vitro replication of feline coronavirus (FCoV), also termed feline infectious peritonitis virus (FIPV). Perylene compounds exhibited high affinity for liposomal and cellular membranes, and efficiently intercalated into the envelopes of SARS-CoV-2 virions, thereby blocking the viral-cell fusion machinery. Furthermore, the studied compounds were demonstrated to be potent photosensitizers, generating reactive oxygen species (ROS), and their anti-SARS-CoV-2 activities were considerably enhanced after irradiation with blue light. Our results indicated that photosensitization is the major mechanism underlying the anti-SARS-CoV-2 activity of perylene derivatives, with these compounds completely losing their antiviral potency under red light. Overall, perylene-based compounds are broad-spectrum antivirals against multiple enveloped viruses, with antiviral action based on light-induced photochemical damage (ROS-mediated, likely singlet oxygen-mediated), causing impairment of viral membrane rheology.

Department of Biology Lomonosov Moscow State University Moscow 119991 Russia

Department of Chemistry and Biochemistry Mendel University in Brno CZ 61300 Brno Czech Republic

Department of Chemistry Lomonosov Moscow State University Moscow 119991 Russia; Shemyakin Ovchinnikov Institute of Bioorganic Chemistry Moscow 117997 Russia

Department of Pharmacology and Toxicology Veterinary Research Institute CZ 621 00 Brno Czech Republic

Laboratory of Emerging Viral Diseases Veterinary Research Institute CZ 621 00 Brno Czech Republic

Laboratory of Emerging Viral Diseases Veterinary Research Institute CZ 621 00 Brno Czech Republic; Department of Experimental Biology Faculty of Science Masaryk University CZ 62500 Brno Czech Republic

Laboratory of Emerging Viral Diseases Veterinary Research Institute CZ 621 00 Brno Czech Republic; Institute of Parasitology Biology Centre of the Czech Academy of Sciences CZ 370 05 České Budějovice Czech Republic; Department of Experimental Biology Faculty of Science Masaryk University CZ 62500 Brno Czech Republic

Laboratory of Emerging Viral Diseases Veterinary Research Institute CZ 621 00 Brno Czech Republic; Institute of Parasitology Biology Centre of the Czech Academy of Sciences CZ 370 05 České Budějovice Czech Republic; Department of Experimental Biology Faculty of Science Masaryk University CZ 62500 Brno Czech Republic; Department of Pharmacology and Pharmacy Faculty of Veterinary Medicine University of Veterinary Sciences Brno CZ 612 42 Brno Czech Republic

Laboratory of Emerging Viral Diseases Veterinary Research Institute CZ 621 00 Brno Czech Republic; Institute of Parasitology Biology Centre of the Czech Academy of Sciences CZ 370 05 České Budějovice Czech Republic; Department of Experimental Biology Faculty of Science Masaryk University CZ 62500 Brno Czech Republic; Faculty of Science University of South Bohemia Ceske Budejovice CZ 37005 Czech Republic

Shemyakin Ovchinnikov Institute of Bioorganic Chemistry Moscow 117997 Russia

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Alkyl Derivatives of Perylene Photosensitizing Antivirals: Towards Understanding the Influence of Lipophilicity

Membrane-Targeting Perylenylethynylphenols Inactivate Medically Important Coronaviruses via the Singlet Oxygen Photogeneration Mechanism