Peptides for treatment of infections of viral origin

Abstract

To provide peptides for treatment of infections of viral origin.SOLUTION: There are provided: peptides that have specific amino acid sequences, as well as pharmaceutical compositions comprising the peptides; and pharmaceutical compositions for the treatment or prevention of infections caused by viruses that infect epithelial cells of the mammalian respiratory system, comprising peptides that have specific amino acid sequences; The invention includes the use of peptides that have specific amino acid sequences for the manufacture of a medicament for the treatment or prevention of infections caused by viruses that infect epithelial cells of the mammalian respiratory system. Also disclosed is a combination of at least one peptide of specific amino acid sequences and an antiviral medication.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to the fields of virology and the pharmaceutical industry. In particular, the present invention discloses a group of synthetic peptides with interrelated sequences that have antiviral activity against infections by a wide range of viruses that infect epithelial cells of the respiratory system, as well as synergistic combinations of the peptides with other antiviral agents. [Background technology]

[0002] Acute respiratory diseases are among the most common illnesses in all age groups. A significant proportion of these diseases are caused by viruses that specifically infect respiratory epithelial cells, either as an initial or subsequent gateway into the respiratory system, in both humans and animals. The airway epithelium possesses morphological and physiological characteristics that distinguish it from the rest of the epithelium. Most airway epithelia are composed of a single layer of cells, but are pseudostratified, with nuclei not aligned. Unlike other epithelial cells, however, they are predominantly ciliated. Mucus-secreting goblet cells are another cell type present in this epithelium but absent in other epithelia. The presence of cilia directed toward the airway lumen, along with the action of goblet cells, maintains humidity within the tract and prevents particles from entering the lungs. Another distinctive feature of respiratory epithelial cells is the presence of numerous pattern recognition receptors (PRRs) that can recognize so-called PAMPs (Pathogen Associated Molecular Patterns), which immediately activate the innate immune response in the presence of pathogens. Ten Toll-like receptors (TLR1 to TLR10) are expressed in the airway epithelium. In particular, TLR3, TLR7, TLR8, and TLR9 constitute a TLR family specialized for detecting viruses infecting the airway. Another characteristic of these cells related to defense functions is their ability to produce molecules with antiviral activity, such as interferons, defensins, and nitric oxide (NO). Furthermore, they can secrete adaptive immune responses that activate cytokines and chemokines (Vareille M et al. 2011, Clin Microbiol Rev, Volume 24, No. 1, pp. 210-229).

[0003] Among the viruses that enter through the respiratory tract are the Orthomyxoviridae family, which includes influenza viruses; the Paramyxoviridae family, which includes parainfluenza, respiratory syncytial virus, and human metapneumovirus; the Picornaviridae family, which includes rhinoviruses and coxsackieviruses; the Adenoviridae family, which includes adenoviruses; and the Coronaviridae family, represented by coronaviruses (Nichols GW et al. 2008 Clin Microbiol Rev, Volume 21, No. 2, pp. 274-290; De Clerk and Li. 2016, Clin Microbiol Rev, Volume 29, pp. 695-747).

[0004] All of the viruses mentioned above cause disease in humans and animals. Some of them, such as seasonal influenza, cause significant economic losses each year due to their high morbidity but low mortality rates. However, several influenza variants generated by adaptation from animal-infecting viruses have caused extremely deadly outbreaks, such as the 1918 Spanish H1N1 influenza (40-100 million deaths), the 1957 Asian H2N2 influenza (1.1 million deaths), or the 1958 Hong Kong H3N2 influenza (500,000 deaths). In recent years, H5N1 avian influenza, first detected in China in 1996, has caused outbreaks in Asia and North Africa with a 50% mortality rate. Respiratory syncytial virus is estimated to result in 3.2 million hospitalizations, 59,600 deaths, and 118,200 deaths annually among children under the age of five (Shi T et al. 2017. Lancet, Volume 390, p. 946-58).

[0005] On the other hand, there are viruses that infect hosts through other routes and initially replicate in other tissues, but can also infect the respiratory tract and cause severe damage during the infection process. Among this second group is the dengue virus, which enters the bloodstream via mosquito bites but can infect epithelial lung cells, causing exacerbation of disease (Cheng NM et al. 2017. PLOS Neglected Tropical Diseases. doi.org / 10.1371 / journal.pntd.0005520; Lee YR et al. 2007, Virus Res, Volume 126, No. 1, pp. 216-25).

[0006] The dengue virus infects approximately 390 million people worldwide each year. Of these, 500,000 develop severe disease or hemorrhagic dengue fever, resulting in 25,000 deaths annually. Indeed, many severe cases of dengue are associated with these respiratory disorders. Some of these pulmonary complications include pulmonary infiltrates, pleural effusion, noncardiogenic pulmonary edema, and respiratory failure (Cheepsattayakorn A and Cheepsattayakorn R. 2014, Journal of Respiratory Medicine Research and Treatment doi: 10.5171 / 2014.162245; Marchiori E et al. 2009, Orphanet Journal of Rare Diseases, Volume 4, No. 8). Other viruses that exhibit these characteristics are herpes simplex virus (HSV) and cytomegalovirus (CMV), both members of the herpesviridae family. HSV causes nosocomial viral pneumonia, bronchopneumonia, and acute respiratory syndrome (Luyt CE et al. 2011, Presse Med, Volume 40, p. E561-e568).

[0007] Antiviral drugs have been developed for some of these viruses, showing comparative therapeutic efficacy. For example, among the antiviral drugs approved for human use are doxuridine, trifluridine, and brivudine for herpes viruses, oseltamivir, zanamivir, and baloxavir marboxil for influenza, and palivizumab and tocozanol for respiratory syncytial virus. Nevertheless, there is a continuing need for the development of new and more effective antiviral drugs with novel mechanisms of action, lower costs, and fewer adverse effects than existing drugs. Therefore, the development of new antiviral agents that act through mechanisms different from existing ones is a global health priority.

[0008] For other viruses, such as dengue virus and coronaviruses, there are no suitable effective treatments (De Clerk and Li, 2016, Clin Microbiol Rev, Volume 29, p. 695-747). In particular, members of the Coronaviridae family are responsible for a wide range of diseases in animals and humans. SARS-CoV, MERS-CoV, and SARS-CoV-2 cause epidemics and severe respiratory infections. The outbreak of the zoonotic virus SARS-CoV (Severe Acute Respiratory Syndrome Coronavirus) in 2003 (Drosten C et al. 2003, New England Journal of Medicine, Volume 348, pp. 1967–1976) and the emergence of MERS-CoV (Middle East Respiratory Syndrome Coronavirus) a decade later (Cui J et al. 2019, Nature Reviews, Volume 17. doi.org / 10.1038 / s41579-018-0118-9) provided the first evidence of the extensive damage caused by this type of virus in humans. Both coronaviruses caused severe respiratory illness and infected nearly 10,000 people before being brought under control.

[0009] This year, a novel coronavirus, SARS-CoV-2, was reported, causing the coronavirus disease 2019 (COVID-19) outbreak (Zhou P et al. 2020, Nature, 579, 270-273, doi.org / 10.1038 / s41586-020-2012-7), which was later declared a COVID-19 pandemic by the World Health Organization. The disease caused by this virus, known as COVID-19, is characterized by an acute respiratory syndrome that can be fatal in approximately 3% of cases. The pandemic has spread rapidly around the world, with over 70 million confirmed cases and over 1.5 million deaths as of November 2020.

[0010] There is little clinical evidence demonstrating the safety and efficacy of any drug against any coronavirus in humans, including SARS-CoV-2 (Kalil AC et al. 2020, JAMA, Volume 323, No. 19, pp. 1897-1898). There is also no clinical evidence of any drug demonstrating antiviral efficacy in patients with persistent SARS-CoV-2 infection lasting longer than 14 days. While remdesivir (Wang Y et al. 2020, Lancet, Volume 395, No. 10236, doi.org / 10.1016 / S0140-6736(20)31022-9) is an antiviral compound that has shown some therapeutic efficacy in clinical trials, its scope is limited to a narrow spectrum of patients, and its therapeutic effect is not as potent as needed. The lack of treatments supported by sufficient scientific evidence has led to the use of different treatment regimens and rapid changes in protocols. The lack of proven treatments and the need for clinical trials to establish evidence-based treatment guidelines have been highlighted (Diaz E et al. 2020, Med Intensiva, doi.org / 10.1016 / j.medin.2020.06.017). There remains an unmet medical need for the treatment of SARS-CoV-2 infection. The search for new antiviral therapeutic options for coronavirus infections, particularly SARS-CoV-2, constitutes an urgent need for human health worldwide. The threat of new pandemics caused by coronaviruses, influenza, or other respiratory viruses necessitates the need for novel antiviral drugs that can control viral infections. Summary of the Invention [Means for solving the problem]

[0011] Description of the Invention The present invention solves the above-mentioned problems by providing peptides having an amino acid sequence selected from the group consisting of SEQ ID NOs: 2 to 20, as well as pharmaceutical compositions comprising at least one of these peptides and a pharmaceutically acceptable excipient. In one embodiment of the present invention, the pharmaceutical composition is formulated for parenteral or mucosal administration. The present invention also provides a pharmaceutical composition for treating or preventing infectious diseases caused by viruses that infect epithelial cells of the mammalian respiratory system, comprising a peptide having the amino acid sequence identified as SEQ ID NO: 1 and a pharmaceutically acceptable excipient. The group of synthetic peptides identified in the Sequence Listing as SEQ ID NOs: 1 to 20 are related to each other by their primary structure.

[0012] In particular, the present invention relates to the inhibition of viral infections caused by viruses that directly infect epithelial cells of the respiratory tract, such as viruses of the Coronaviridae, Orthomyxoviridae and Paramyxoviridae, Picornaviridae, and Adenoviridae families. More specifically, the present invention relates to influenza, parainfluenza, respiratory syncytial virus, coronavirus, enterovirus, and adenovirus, particularly SARS-CoV-1, SARS-CoV-2, MERS-CoV, bovine coronavirus (BCoV), influenza A, parainfluenza type 3, H1N1, H5N1, adenovirus Ad7, and bocavirus HBoV-1. The present invention also relates to the treatment of viral infections caused by dengue and Zika viruses, herpes simplex viruses, and cytomegalovirus, avoiding or treating the serious damage these viruses can cause in the respiratory tract, primarily at the lung level. Viruses that primarily affect animals, such as swine fever virus, are also included.

[0013] The peptide identified as SEQ ID NO:1 has previously been reported as an inhibitor of HIV replication through modifications in the vimentin cytoskeleton that prevent viral replication in cells in which the cytoskeletal intermediate filaments are formed by vimentin (Fernandez-Ortega C et al. 2016, Viruses, Volume 8, No. 6, doi.org / 10.3390 / v8060098). Meanwhile, prior art studies have shown that vimentin is an important target for viral inhibition in cells with cytoskeletal intermediate filaments formed by this protein. In 2015, Zhang et al. reported that vimentin gene silencing affected viral replication of TGEV porcine coronavirus (Zhang X et al. 2015, Virus Research, http: / / dx.doi.org / 10.1016 / j.virusres.2014.12.013).

[0014] More recently, it has been reported that vimentin phosphorylation is regulated in cells infected with SARS-CoV-2 (Bouhaddou M et al. 2020, Cell, volume 182, pp. 685-712. https: / / doi.org / 10.1016 / j.cell.2020.06.034). However, in the present invention, the peptide identified in the Sequence Listing as SEQ ID NO: 1 was found to exhibit strong antiviral activity against coronaviruses in cells that do not display the vimentin cytoskeleton, the primary target of viral infection. Furthermore, it was demonstrated that the antiviral activity of this peptide, which is useful for treating viral infections referred to in the present invention, is independent of the vimentin cytoskeleton. More specifically, the inventors surprisingly demonstrate that this peptide can be used to treat diseases caused by viruses that infect cells for which those skilled in the art would not expect any effect. The present invention also includes undescribed peptides that have strong antiviral activity against viruses that infect epithelial cells of the respiratory system.

[0015] The first and second examples of the present invention demonstrate the antiviral activity of the peptide identified as SEQ ID NO: 1 and other related peptides against BCoV and SARS-CoV-2 in cell lines that display vimentin intermediate filaments as part of their cytoskeleton. Surprisingly, these peptides do not affect the reorganization of the vimentin cytoskeleton in these cells. Although these cell lines are of epithelial origin, they developed a vimentin cytoskeleton in vitro. While the state of the art would predict its modification by the action of the peptides, this does not occur. Despite the fact that these cells possess vimentin filaments, the peptides of the present invention exhibit inhibitory activity against viral cytopathic effects without modifying these cytoskeletal filaments. Even more surprising is the demonstration that the peptides identified as SEQ ID NOs: 1 to 20 can inhibit infection by coronaviruses and other viruses in primary cultures of respiratory epithelial cells that do not display vimentin intermediate filaments as part of their cytoskeleton.

[0016] Meanwhile, previous studies have reported direct interaction between vimentin and the spike (S) protein of SARS-CoV during viral entry. Furthermore, vimentin is known to play a role in the binding of SARS-CoV to its cellular receptor and viral entry into cells (Yu et al. 2016, Journal of Biomedical Science, volume 23, number 14. doi 10.1186 / s12929-016-0234-7). Surprisingly, the peptides of the present invention do not interact with either the S protein of SARS-CoV-2 or its ACE2 receptor, but rather with the N protein of the virus.

[0017] Another unexpected result of the present invention is that viral inhibition caused by the peptides identified in the Sequence Listing as SEQ ID NOS: 1-20 was not achieved in all epithelial cells. In the case of the HSV-1 virus, its replication is efficiently inhibited by the described peptides in primary cells of the nasal mucosa. However, examples of the present invention show that inhibition of viral replication was not achieved in epithelial cells other than those of the respiratory tract. Taken together, all these results demonstrate, quite surprisingly, that these peptides have antiviral activity against viruses that infect respiratory epithelial cells. This finding refers to, but is not limited to, the inhibition of replication of SARS-CoV-2, BCoV, influenza A H1N1, parainfluenza type 3, respiratory syncytial virus, adenovirus Ad7, bocavirus HBoV-1, dengue virus 2, herpes simplex virus HSV-1, and swine fever virus.

[0018] Therefore, the antiviral effect of the peptides of the present invention is exerted by a mechanism that does not involve alteration of intermediate vimentin filaments or blocking of virus binding to its receptor on the target cell membrane. This finding confirms that the peptides of the present invention exert their antiviral activity through a novel mechanism not reported in the state of the art. Surprisingly, they are able to inhibit viral infection in epithelial cells of the respiratory system.

[0019] In one embodiment of the present invention, pharmaceutical compositions comprising peptides identified in the Sequence Listing as SEQ ID NOs: 1 to 20 are formulated to be administered parenterally or via mucosal routes. In one embodiment of the present invention, the infections treated with the peptides of SEQ ID NOs: 1 to 20 are caused by viruses from the Coronaviridae, Orthomyxoviridae and Paramyxoviridae, Picornaviridae, and Adenoviridae families. In certain embodiments, pharmaceutical compositions comprising peptides of SEQ ID NOs: 1 to 20 are useful for the treatment or prevention of infections caused by coronavirus, influenza virus, parainfluenza, respiratory syncytial virus, adenovirus, dengue virus, herpes simplex virus, cytomegalovirus, and swine fever virus.

[0020] Another object of the present invention is the use of a peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 20 for the manufacture of a medicament for treating or preventing an infection caused by a virus that infects epithelial cells of the mammalian respiratory system. In a particular embodiment, the medicament comprises two or more peptides having an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 20.

[0021] Thus, the present invention includes the use of the peptide of SEQ ID NO: 1 in the treatment of COVID-19 and other infectious diseases caused by viruses that infect epithelial cells of the respiratory system. The present invention also provides new peptides structurally related to the peptide of SEQ ID NO: 1 for the treatment of these infectious diseases, including those caused by viruses whose initial target of entry is not cells of the respiratory system, but which may have more severe clinical consequences on cells of this system. These viruses include herpesviruses, dengue viruses, and cytomegaloviruses.

[0022] In another aspect, the present invention provides a method for treating or preventing an infection caused by a virus that infects epithelial cells of the mammalian respiratory system, comprising administering to an individual in need thereof a therapeutically effective amount of at least one peptide having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 20. In certain embodiments of the methods of the present invention, the infection is caused by a virus of the Coronaviridae, Orthomyxoviridae and Paramyxoviridae, Picornaviridae, and Adenoviridae families. In preferred embodiments of the methods of the present invention, the infection is caused by a virus selected from the group consisting of coronavirus, influenza virus, parainfluenza, respiratory syncytial virus, adenovirus, dengue virus, herpes simplex virus, cytomegalovirus, and swine fever virus.

[0023] In one embodiment of the present invention, an antiviral drug is further administered to the individual as part of a method of treatment or prevention. The peptide having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 20 and the antiviral drug can be administered simultaneously or sequentially in the same treatment schedule. In one embodiment of the present invention, the antiviral drug is selected from the group consisting of ribavirin, ivermectin, penciclovir, nitazoxanide, nafamostat, remdesivir, favipiravir, the peptide identified as SEQ ID NO: 23, interferon (IFN) alpha, IFN gamma, or a combination thereof.

[0024] Another object of the present invention is a pharmaceutical combination comprising two or more peptides having an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 20. In one embodiment of the present invention, in the pharmaceutical combination, the peptides are administered simultaneously or sequentially in the course of the same treatment.

[0025] The present invention further contemplates a pharmaceutical combination for treating or preventing infections caused by viruses that infect epithelial cells of the mammalian respiratory system, characterized in that it comprises: a) at least one peptide having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 20; and b) an antiviral drug. In a specific embodiment, the antiviral drug in the pharmaceutical combination is selected from the group formed by ribavirin, ivermectin, penciclovir, nitazoxanide, nafamostat, remdesivir, favipiravir, the peptide identified as SEQ ID NO: 23, IFN-alpha, IFN-gamma, or a combination thereof. In such a pharmaceutical combination, the peptide and the antiviral drug are administered simultaneously or sequentially during the same course of treatment.

[0026] The present invention demonstrates that the combination of peptides of SEQ ID NO: 1 to SEQ ID NO: 20 with other antiviral compounds enhances their inhibitory activity, resulting in synergistic antiviral activity. The antiviral drug is selected from the group formed by ribavirin, ivermectin, penciclovir, nitazoxanide, nafamostat, remdesivir, favipiravir, the peptide identified as SEQ ID NO: 23, interferon alpha, interferon gamma, or a combination thereof, but the scope of the present invention is not limited thereto. [Brief explanation of the drawings]

[0027] [Figure 1] Inhibition of replication of the Mebus strain of bovine coronavirus by the peptide identified in the Sequence Listing as SEQ ID NO: 1. (A): Inhibition of cytopathic effect; (B): Reduction of viral ribonucleic acid (RNA) copy number by RT-qPCR. [Figure 2] Immunodetection of vimentin intermediate filaments in cell lines. MDBK cells (Panel A) and VeroE6 cells (Panel B) were treated with the peptide identified as SEQ ID NO: 1 in the Sequence Listing at 100 μM for 24 hours. No difference in the structure of vimentin filaments was observed between treated and untreated cells. CC: untreated cells; Target: vimentin cytoskeleton; 40x magnification. [Figure 3]Sensorgrams corresponding to the interaction assay of different molecules with the peptide identified in the Sequence Listing as SEQ ID NO: 1. (A) SARS-CoV-2 protein N, (B) Dengue 2 virus capsid protein, (C) HIV p24 protein, (D) polyclonal anti-SEQ ID NO: 1. All analytes were measured in the concentration range of 10 nM to 200 nM, and at least five curves per sample were taken into account to estimate the characteristic parameters of the interaction. [Figure 4] Inhibition of swine fever virus by the peptide identified in the Sequence Listing as SEQ ID NO: 1. A. Inhibitory effect of viral replication in PK-15 cells. B. Cytotoxicity of the peptide in PK15 cells assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazole (MTT) bromide assay. [Figure 5] Determination of the antiviral effect of combined treatment with the peptide identified in the Sequence Listing as SEQ ID NO: 1 and interferon alpha 2b. MDBK cells were treated 24 hours prior to infection with the Mebus strain of bovine coronavirus (BCO). The antiviral effect was assessed by determining the infectivity of the virus present in the culture supernatant 72 hours after infection. DETAILED DESCRIPTION OF THE INVENTION

[0028] example Example 1. Inhibition of bovine coronavirus replication by the peptide identified as SEQ ID NO:1. The peptide identified as SEQ ID NO:1 in the Sequence Listing was obtained by chemical synthesis using solid-phase synthesis. To evaluate the antiviral effect of this peptide, bovine kidney cells (MDBK), also known as MDBK, were seeded at 280,000 cells per well in a 24-well plate in DMEM culture medium containing 10% fetal bovine serum (FBS). The plates were incubated at 37°C and 5% CO2 in a humidified atmosphere. After 18 to 24 hours, different concentrations of the peptide identified as SEQ ID NO:1 were added and incubated under the same conditions for 1 hour or 24 hours. After this treatment period, the culture medium was removed, and the cells were infected with the Mebus strain of bovine coronavirus at a multiplicity of infection (MOI) of 0.01 in 200 μL of serum-free DMEM medium for 1 hour in a humidified atmosphere at 37°C and 5% CO2. The volume of each well was then adjusted to 1 mL with serum-free DMEM medium in the presence of the peptide. After 120 hours of incubation, the viability of the cell cultures was assessed by the MTT metabolic degradation method. To do this, the culture medium was removed. 200 microliters of MTT was added to each well, and the plate was incubated at 37°C, 5% CO2, and 95% humidity for 2 to 4 hours. Then, 300 μL of isobutyl alcohol was added, and the plate was shaken for 1 hour. As an alternative to the MTT method, cells were stained with crystal violet (0.1%). Finally, the absorbance of each well was measured at 570 nm in a plate reader (Amersham). The antiviral effect was expressed as the percentage of inhibition of cytopathic effect (CPE) (Figure 1A) and viral RNA by RT-qPCR (Figure 1B). The results demonstrate that the peptide identified as SEQ ID NO: 1 exerts an antiviral effect against bovine coronavirus infection.

[0029] Example 2. Evaluation of the antiviral effect of peptides identified as SEQ ID NO: 1 to SEQ ID NO: 22 against SARS-CoV-2 virus. The peptides identified as SEQ ID NOs: 2 to 22 in the Sequence Listing were chemically synthesized using solid-phase synthesis. To evaluate the antiviral effects of the peptides identified as SEQ ID NOs: 1 to 22 against SARS-CoV-2 coronavirus, VeroE6 cells (African green monkey kidney cells) were seeded in 96-well plates at 20,000 cells per well in MEM culture medium containing 10% FBS and incubated at 37°C and 5% CO2 in a humidified atmosphere. After approximately 24 hours, the peptides to be evaluated were added at concentrations ranging from 0.01 μM to 100 μM and incubated under the same conditions for 1 hour or 24 hours. The culture medium was then removed, and the cells were infected with SARS-CoV-2 coronavirus at an MOI of 0.01 in 100 μL of MEM culture medium containing 2% FBS for 1 hour in a humidified atmosphere at 37°C and 5% CO2. The volume of each well was then adjusted to 200 μL with MEM culture medium containing 2% FBS in the presence of the peptides. After 96 hours of incubation, cell viability in the cultures was assessed by neutral red staining. To do this, the medium was removed, 100 μL of dye was added, and the cells were incubated for 1 hour at 37°C, 5% CO2, and 95% humidity. The absorbance was measured at 580 nm in a plate reader. The antiviral effect was expressed as the percentage inhibition of CPE. The results demonstrate that the peptide identified as SEQ ID NO: 1 and several related peptides exert antiviral effects against bovine coronavirus infection, as observed in Table 1. [Table 1]

[0030] Example 3. Evaluation of the effect of the peptide identified as SEQ ID NO: 1 on vimentin intermediate filaments in MDBK and VeroE6 cells. To evaluate the effect of the peptide identified as SEQ ID NO:1 on vimentin intermediate filaments, MDBK and Vero E6 cells were treated with 100 μM of the peptide at 37°C and 5% CO2 for 24 hours. They were then fixed, permeabilized in a solution of acetone:methanol (1:1 v / v) for 10 minutes at -20°C, and blocked with 1% bovine serum albumin in phosphate-buffered saline for 30 minutes at 37°C. Vimentin intermediate filaments were detected using a mouse anti-vimentin IgG1 monoclonal antibody (4.5 μg / mL) followed by an anti-mouse antibody conjugated to fluorescein isothiocyanate (1 / 200). Vimentin intermediate filaments (FI) were observed distributed throughout the cytoplasm, from the perinuclear region toward one or both poles of the cell. No differences in supramolecular structure were observed between treated and untreated cells under the conditions evaluated. No damage to cell integrity was observed after treatment. The vimentin cytoskeleton was not altered by the peptide identified as SEQ ID NO: 1 in MDBK cells (Figure 2A) or VeroE6 cells (Figure 2B). In similar experiments performed with peptides of SEQ ID NOs: 2-22, no alterations were observed in the structure of intermediate filaments.

[0031] Example 4. Anti-SARS-CoV-2 activity exhibited by various peptides in primary cultures of respiratory epithelial cells We used primary epithelial cells obtained from the respiratory tract to determine whether the peptides under investigation inhibit SARS-CoV-2 viral replication in cells that lack the vimentin cytoskeleton. These cells are known to lack vimentin intermediate filaments (Robinson-Bennett and Han A 2006. Handbook of Immunohistochemistry and In Situ Hybridization of Human Carcinomas, vol. 4. Edited by MA Hayat; Elsevier. pp. 537-47). The antiviral activity of peptides identified in the Sequence Listing as SEQ ID NOS: 1-22 against SARS-CoV-2 coronavirus was evaluated in human airway epithelium (HAE) obtained from nasal or bronchial biopsies or organ donors according to Fulcher et al. (Fulcher et al. 2003, Methods in Molecular Medicine, vol. 107, Second Edition, Ed: J. Picot (c) Humana Press Inc., Totowa, NJ). Primary cells were obtained with individual consent.

[0032] Primary cultures of human airway epithelium were maintained at the air-liquid interface in Transwell plates and infected with SARS-CoV-2 at an MOI of 1 for 1 h at 37°C and 5% CO2. Cultures were treated with peptides ranging from 0.01 μM to 100 μM for 1 h or 24 h before virus challenge. After virus challenge, peptides were substituted in the cultures every 24 h. After 72 h, apical washes and SARS-CoV-2 RT-qPCR were performed. Strikingly, the peptide inhibited replication of this virus in cells that do not display intermediate vimentin filaments, indicating an independent antiviral mechanism for this protein (Table 2). [Table 2]

[0033] The remaining peptides identified in the Sequence Listing as SEQ ID NO: 1 to SEQ ID NO: 22 were also evaluated in the system as described, and they inhibited SARS-CoV-2 in primary cultures of human airway epithelium, similar to Table 2, with an inhibition range between 60-70%, except for peptides SEQ ID NO: 21 and 22, which did not achieve an inhibition rate above 20%.

[0034] Example 5. Interaction of peptides with SARS-CoV-2 protein N and other viral proteins The unexpected results, which indicated that the inhibitory effect of these peptides does not occur via the cytoskeleton, led us to explore possible pathways of action. In particular, we investigated the interaction of the peptides with SARS-CoV-2 proteins, as well as with several proteins involved in the virus's entry into human host cells, including the nucleocapsid protein (N protein) of the SARS-CoV-2 virus, the extracellular domain of the ACE2 receptor, and the receptor-binding domain (RBD) of the viral S protein. Similarly, we investigated the interaction of the peptides with proteins from other viruses, as described below. In this study, we used a BIACORE-type sensor, which allows us to characterize the interaction between two molecules based on an optical principle called surface plasmon resonance. To perform the required measurements, a biotinylated peptide identified as SEQ ID NO: 1 (under medium-density conditions) was immobilized on both channels of a streptavidin (SA)-functionalized chip (General Electric Healthcare, USA). After a regeneration process, the fc1 channel was used as the sample channel, and the fc2 channel was used as the reference channel to confirm the integrity of the immobilized molecules on the surface.

[0035] To assess the reactivity of the immobilized peptide, a specific polyclonal antibody obtained in rabbits at a titer of 1 / 3000 was used against the peptide identified as SEQ ID NO: 1. Application of the polyclonal antibody caused an increase in signal (RU), evidence of molecular recognition that can be correlated with its integrity after immobilization on the chip surface (Figure 3D, Table 3).

[0036] Analysis of the interaction of the peptide identified as SEQ ID NO: 1 with different proteins revealed that neither the RBD nor the ACE2 peptide interacts with the peptide identified as SEQ ID NO: 1. Protein N showed a significant increase in signal (RU) upon interaction with the immobilized peptide identified as SEQ ID NO: 1 in the concentration range examined (10-200 nM) (Figure 3A). The estimated K for this interaction was D is 3.58 x 10 -9 M, which assumes a high affinity of interaction (in the nanomolar range) between both interactants.

[0037] Figure 3B shows a sensorgram obtained from a study of the interaction of the peptide identified as SEQ ID NO: 1 with the Dengue 2 virus capsid protein, where a concentration-dependent interaction of the protein can be observed. On the other hand, the sensorgram of the interaction of the peptide identified as SEQ ID NO: 1 with the HIV-1 p24 protein (Figure 3C) shows no interaction. Other viral proteins with similar characteristics to SARS-CoV-2 protein N, such as the envelope protein of Zika virus (ZIKV E), the nonstructural protein 1 (NS1-D2) of Dengue virus 2, and the capsid protein of influenza H1N1 virus, showed interactions with the peptides identified as SEQ ID NOs: 1, 2, 14, and 15. The most important parameters characterizing the interaction of this peptide with various viral proteins are listed in Tables 3 and 4.

[0038] In the case of the HIV-1 p24 protein, there was no interaction with the peptides evaluated (Figure 3C). This result confirms that the inhibition of HIV-1 infection by the known peptide identified as SEQ ID NO: 1 is caused by a different mechanism of action of this peptide under specific conditions that are not met in epithelial cells of the respiratory system. [Table 3]

[0039] Surprisingly, all these results point to the fact that the antiviral activity of the peptides of the invention is caused by a direct mechanism of interaction with the virus, independent of the host cell cytoskeleton. [Table 4]

[0040] The remaining peptides identified in the Sequence Listing as SEQ ID NO: 1 to SEQ ID NO: 20 were evaluated with similar results. The peptides identified as SEQ ID NO: 21 and SEQ ID NO: 22 did not interact with the proteins evaluated.

[0041] Example 6. Inhibition of swine fever (CSF) virus by the peptide identified in the Sequence Listing as SEQ ID NO: 1. To evaluate the antiviral effect of the peptide identified as SEQ ID NO: 1 in the Sequence Listing, 8500 PK15 cells per well were seeded in 50 μL of DMEM medium in a 96-well plate, and either 50 μL of DMEM or the peptide diluted in DMEM was added. Starting at 80 μM, two-fold serial dilutions of the peptide were made. Each point was evaluated in triplicate, and three virus-free cell controls were included. The plates were incubated at 37°C and 5% CO2 for 24 hours. 100 TCID50 of the CSF virus, Margarita strain, was added per well (MOI 0.01). The plates were incubated for 72 hours, the medium was discarded, and the cells were fixed at 80°C for 1 hour. They were then blocked with 100 μL of 2% milk in PBS at 37°C for 1 hour. They were washed three times with PBS + 0.05% Tween 20, and 100 μL of a 1:3000 dilution of a monoclonal antibody against the E2 viral protein conjugated to peroxidase was added and incubated for 1 h at 37°C. The plates were washed five times with PBS + 0.05% Tween 20, and substrate was added (10 mL of citrate buffer pH 5.0 containing 5 mg of orthophenylenediamine + 5 μL of H2O2). After a 10-minute incubation at room temperature, the optical density (OD) was determined at 492 nm in an ELISA plate reader.

[0042] Inhibitory concentration 50 (IC 50 To obtain α- and β-actin values, corresponding dose-response curves were generated (Figure 4), and these values ​​were estimated using GraphPad Prisma software. In parallel, the cytotoxic effect of the peptides on PK15 cells was tested by MTT assay in the concentration range of 1 μM to 400 μM. For this, incubation with the peptides was extended to 96 h, followed by color development by treatment with MTT.

[0043] The results obtained demonstrate that the peptide identified as SEQ ID NO: 1 is able to exert a concentration-dependent inhibitory effect on CSF viral replication (Figure 4A) in a dose range that does not affect cell viability (Figure 4B). 50 (300 nM) is the cytotoxic concentration of 50 (CC 50 These peptides were also shown to give a very favorable safety index (SI) of 1333, much higher than that of the 1333-like peptides (>400 μM). The remaining peptides identified in the Sequence Listing as SEQ ID NO: 2 to SEQ ID NO: 20 were also evaluated in the described system with similar results.

[0044] Example 7. Demonstration of the antiviral effect of peptides against various viruses that infect the respiratory tract. Primary cultures of HAE were obtained from the nasal passages, trachea, bronchi, and / or lungs of organ donors according to Fulcher et al. (Fulcher et al. 2003, Methods in Molecular Medicine, vol. 107, Second Edition. Ed: J. Picot (c) Humana Press Inc., Totowa, NJ). Inhibition studies were performed with different respiratory viruses using different panels of peptides of the invention to facilitate initial validation.

[0045] To evaluate the antiviral activity of peptides identified as SEQ ID NOS: 1, 2, 3, 7, 9, 14, and 19 against influenza H1N1 virus, HAEs derived from tracheobronchial tissue were cultured in 12- or 24-well Transwell plates (Costar) and pretreated with peptides at concentrations ranging from 0.01 to 100 μM for 2 or 24 hours before virus challenge. The cultures were infected with influenza A H1N1 at 100 PFU (plaque-forming units) for 1 hour at 4°C, washed with PBS, and incubated at 37°C and 5% CO2 for 24 hours. Apical washes were then performed using the collected PBS to evaluate viral potency in MDCK cells. To do this, MDCK cells were seeded in 96-well plates, and when they reached confluence, serial 10-fold dilutions of the apical washes were used to infect the cells. The cells were then incubated at 37°C for 1 hour. Next, they were washed with PBS, and an agar layer in 2x DMEM (1:1) was added and allowed to solidify. Plates were then incubated at 37°C, 5% CO for 48 hours and stained with crystal violet to determine PFU numbers. The peptides inhibited viral replication by percentages ranging from 67 to 93 (Table 5).

[0046] To evaluate the antiviral activity of peptides identified as SEQ ID NOS: 1, 2, 8, 13, 15, and 20 against parainfluenza virus type 3 (HPIV3), the assay was performed as described for influenza H1N1, except that infection was performed with 4000 PFU at 37°C for 90 min. After washing with PBS, cultures were incubated in the presence of peptides for 3 days. Afterward, apical washes with PBS were performed, harvested, and viral potency in MDCK cells was evaluated, as described previously. Results show percentages of viral inhibition between 73 and 90% in the presence of peptides at the concentrations evaluated (Table 5).

[0047] The antiviral activity of peptides identified as SEQ ID NOs: 1, 2, 3, 13, 15 and 20 against respiratory syncytial virus (RSV) was tested as described for influenza H1N1 but at 2×10 6Infection with PFU was performed for 2 hours at 37°C and assessed in HAEs obtained from the bronchi. After washing with PBS, cultures were incubated for 10 days, with medium and peptides being refreshed every 2 days. Virus was assessed by detecting RSV nucleoprotein RNA by RT-qPCR. Viral inhibition values ​​reached >75% (75-91%) (Table 5).

[0048] To evaluate the effects of peptides identified as SEQ ID NOS: 1, 3, 7, 9, 13, and 15 against adenovirus type 7, we infected HAE cultures obtained from bronchi with the AdV7 Gomen strain at an MOI of 1 for 1 h at 37°C, as described for influenza H1N1 virus. Cultures were washed with PBS and incubated for 24 h at 37°C and 5% CO2. Apical lavage was then performed with PBS, which was collected for virus detection by qPCR. Viral inhibition values ​​in the presence of peptides reached greater than 68% (range, 68-82%) (Table 5).

[0049] The efficacy of peptides identified as SEQ ID NOs: 3, 8, 9, 19, and 20 against human bocavirus (HBoV) was achieved and measured as described for H1N1, but using tracheal-derived HAE. Cultures were infected at an MOI of 1 for 2 h at 37 °C. The medium was then removed, and cultures were maintained for 72 h before apical washes were performed and the presence of virus in the NS1 region was quantified by qPCR. Viral inhibition values ​​obtained in the presence of peptides ranged from 67 to 87% (Table 5).

[0050] To evaluate the inhibition of dengue virus replication by peptides identified as SEQ ID NOS: 1, 2, 7, 8, 14, and 19, primary cultures of lung cells obtained from lung cancer patients and maintained in DMEM culture medium containing 30% FBS were performed. Prior to viral challenge, the cultures were treated with peptides at concentrations ranging from 0.01 μM to 100 μM for 2 or 24 hours. The medium was discarded, and the cells were infected with Dengue 2 virus at an MOI of 10 for 2 hours at 37°C and 5% CO2. After virus removal and subsequent washing, the cultures were treated again with peptides in DMEM medium and incubated for 48 hours at 37°C and 5% CO2. The cultures were then harvested to evaluate viral efficacy against VeroE6 cells. For this purpose, cells were seeded into 24-well plates, and upon reaching confluence, they were infected with serial 10-fold dilutions of the supernatant and incubated for 1 hour at 37°C and 5% CO2. They were then washed with PBS, and an agar layer in 2x DMEM (1:1) was added. The layers were allowed to solidify and incubated for 48 hours at 37°C and 5% CO2. Plates were stained with crystal violet to determine the number of PFU. The peptides inhibited viral replication by 68-88% (Table 5).

[0051] Herpes simplex virus type 1 (HSV-1) replication was performed in primary explants of human nasal mucosa according to Glorieux et al. (Glorieux et al. 2011, PLoS ONE, 6(7): e22160, doi: 10.1371 / journal.pone.0022160). Explants were cultured in 24-well plates and treated with peptides identified as SEQ ID NOs: 1, 3, 13, 15, and 20 at concentrations of 0.01 to 100 μM for 2 hours, followed by incubation in serum-free medium for 10 min. 7 TCID 50 Cultures were infected with HSV-1 / mL for 1 hour at 37°C and 5% CO2. Cultures were then washed and maintained for 36 hours at 37°C and 5% CO2 in the presence of peptides in the culture medium. Cultures were harvested, and HSV-1 was quantified by qPCR. The percentage of viral inhibition ranged from 70 to 92% in the presence of peptides (Table 5).

[0052] All peptides identified in the Sequence Listing as SEQ ID NOs: 1-20 were evaluated in subsequent experiments against each of the viruses mentioned, with ranges of inhibition similar to those shown in Table 5. [Table 5]

[0053] Example 8. Effect of peptides on epithelial cells other than those from the respiratory system. To determine whether the peptides of the present invention inhibit viral infection in primary epithelial cells other than those of respiratory origin, HSV-1 infection was tested in primary cultures of human corneal epithelial cells. Cells were seeded in 48-well plates until they reached 80-90% confluence, and they were treated with peptides identified as SEQ ID NOs: 1, 13, and 20 at different concentrations for 2 hours. They were then incubated for 10 min in serum-free medium. 7 TCID50 of HSV-1 / mL was infected for 1 hour at 37°C and 5% CO2. Cells were washed, treated again with peptides, and maintained at 37°C and 5% CO2 for 72 hours. The presence of HSV-1 in the cultures was determined by qPCR. As demonstrated in Example 7, despite the fact that these peptides were able to inhibit this virus in primary respiratory cells, the percentage of virus inhibition obtained in the presence of the peptides was less than 25%.

[0054] Example 9. Synergistic effect of the peptide identified as SEQ ID NO: 1 with alpha interferon in inhibiting bovine coronavirus replication. The antiviral effect of the peptide identified as SEQ ID NO: 1 in combination with recombinant alpha 2b interferon (Heberon™ Alfa R, Cuba) was evaluated in an infection system of the Mebus strain of bovine coronavirus in MDBK cells using a 96-well plate. Peptide treatment was performed at 10 μM for 24 hours, followed by incubation with interferon at 20 μg / mL for 24 hours. The results are shown in Figure 5. As can be seen, the combination of both molecules reduced viral infection approximately 50-fold compared to the peptide alone and 100-fold compared to interferon. The test results demonstrate the enhancement of the antiviral effect of the peptide by co-treatment with interferon. Similar results were obtained with the peptides identified in the Sequence Listing as SEQ ID NO: 2 to SEQ ID NO: 20.

[0055] Example 10. Effect of peptides in combination with different compounds with antiviral activity. To evaluate the effect of the peptide identified as SEQ ID NO: 1 in combination with compounds with antiviral activity against SARS-CoV-2 or other viruses, a SARS-CoV-2 virus challenge assay was performed on VeroE6 cells as described in Example 2. Monolayers of VeroE6 cells were incubated with the peptide identified as SEQ ID NO: 1 at 10 μM for 1 hour or 24 hours, alone or with each antiviral compound at concentrations ranging from 0.1 μM to 100 μM, at 37°C and 5% CO2. Antiviral efficacy was expressed as a percentage of CPE inhibition and was calculated based on the inhibitory concentration 50 (IC50) of each compound in the presence or absence of the peptide. 50 ) was calculated. In Table 6, compounds evaluated in the presence of peptides identified as SEQ ID NOs: 1, 2 and 3 show an enhancing effect on antiviral activity. [Table 6]

[0056] The remaining peptides identified in the Sequence Listing as SEQ ID NO:2 to SEQ ID NO:20 also enhanced the activity of the compounds listed in Table 6.

[0057] Example 11. A pharmaceutical composition comprising one or more of the peptides identified in the Sequence Listing as SEQ ID NOs: 1-20. To obtain a pharmaceutical composition for the prevention or treatment of infectious diseases caused by viruses affecting epithelial cells of the respiratory system, a formulation containing at least one of the peptides identified in the Sequence Listing as SEQ ID NOS: 1-20 at a concentration of 1.0-20.0 mg / mL, sucrose at a concentration between 10.0-20.0 mg / mL, and acetic acid at a concentration between 0.1-6 μg / mL was developed. The composition further contained sodium hydroxide and water for injection in appropriate amounts (sufficient concentrations). The formulations were subjected to accelerated stability testing, and the results showed that they remained as homogeneous white lyophilizates with a purity greater than 95% and a microbial limit of less than 0.01 cfu / mg (colony forming units / mg), were pyrogen-free, had a humidity percentage of less than 10%, a peptide content of more than 60%, and a pH between 2 and 6.

[0058] Example 12. Antiviral effect of the peptide identified as SEQ ID NO: 1 in patients infected with SARS-CoV-2. The human study was conducted in accordance with the Declaration of Helsinki. Patients infected with SARS-CoV-2 were treated with a peptide identified as SEQ ID NO: 1, obtained with quality and adapted for clinical use in humans. The peptide was administered subcutaneously. The presence of SARS-CoV-2 viral RNA was determined by RT-qPCR (Roche) in nasopharyngeal swabs taken from the patients every 48 or 96 hours for 15 days. Four groups were formed, each containing 20 patients with similar characteristics and in the early stages of disease. Patients in Group 1 received the most frequently used treatment for COVID-19, based on Kaletra (lopinavir / ritonavir), chloroquine, and Heberon™ Alpha R (IFNα2b). Patients in Group 2 received this treatment plus the peptide identified as SEQ ID NO: 1 in the Sequence Listing. The latter was administered subcutaneously at 2 mg / kg body weight for 5 days. Patients in Group 3 received treatment based on Kaletra, chloroquine, and a combination formulation of intramuscular IFNα2b (3 million international units, MIU) and IFNγ (0.5 MIU). Patients in Group 4 received the same treatment as Group 3, plus the peptide identified in the Sequence Listing as SEQ ID NO: 1 (2 mg / kg body weight) subcutaneously for 5 days.

[0059] The results showed that patients treated with peptides became SARS-CoV-2 negative more quickly than patients treated similarly without peptides. The number of patients testing negative for SARS-CoV-2 was higher in the groups receiving peptide antiviral treatment (Table 7). Four days after the start of treatment, 80% and 85% of patients treated with peptides (Groups 2 and 4, respectively) were SARS-CoV-2 negative, a higher percentage than the percentage reached by groups treated without peptides (Groups 1 and 3). Six days after the start of treatment, 100% of patients in Groups 2 and 4 were SARS-CoV-2 negative, while patients not treated with peptides reached 100% by Day 14 (Table 7). [Table 7]

[0060] Example 13. Effect of the peptide identified in the Sequence Listing as SEQ ID NO: 1 in healthy individuals in contact with COVID-19 patients. Healthy individuals (contacts of COVID-19 patients) were included in this study. Two groups (Group I and Group II) of 25 healthy individuals exposed to similar risk infectious agents were analyzed. Group II was treated daily for 6 days with the peptide identified in the Sequence Listing as SEQ ID NO: 1 as intranasal drops at a concentration of 14 μg / mL. Nasopharyngeal swab samples were collected for SARS-CoV-2 detection by RT-qPCR (Roche). Sampling was performed at the start of the study and 7 days after the start. The results of SARS-CoV-2 determination showed that individuals treated with the peptide identified in the Sequence Listing as SEQ ID NO: 1 were protected from viral infection, with only one of the 25 individuals tested infected (3.9%), in contrast to the untreated group, in which 12 of the 25 individuals tested (48%) were infected with SARS-CoV-2 (Table 8). [Table 8] [Sequence List Free Text]

[0061] Sequence Listing 1 <223> Description of artificial sequences: synthetic peptides Sequence Listing 2 <223> Description of artificial sequences: synthetic peptides Sequence Listing 2 <223> Acetylation Sequence Listing 3 <223> Description of the artificial sequence: A synthetic peptide that is acetylated and has a D amino acid at N6 Sequence Listing 3 <223> Acetylation Sequence Listing 4 <223> Description of the artificial sequence: A synthetic peptide that is acetylated and has a D amino acid at N8 Sequence Listing 4 <223> Acetylation Sequence Listing 5 <223> Description of the artificial sequence: A synthetic peptide that is acetylated and has a D amino acid at N9 Sequence Listing 5 <223> Acetylation Sequence Listing 6 to Sequence Listing 18 <223> Description of artificial sequences: synthetic peptides Sequence Listing 19 <223> Description of artificial sequences: synthetic peptides where N6 is a D amino acid Sequence Listing 20 <223> Description of artificial sequences: synthetic peptides where N8 is a D amino acid Sequence Listing 21~Sequence Listing 22 <223> Description of artificial sequences: synthetic peptides Sequence Listing 23 <223> Description of an artificial sequence: A synthetic antiviral peptide with a disulfide bond between C15 and C25

Claims

1. A peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 2 to SEQ ID NO:

20.

2. A pharmaceutical composition comprising at least one peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 2 to SEQ ID NO: 20, and a pharmaceutically acceptable excipient.

3. 3. The pharmaceutical composition of claim 2, further comprising a peptide having the amino acid sequence identified as SEQ ID NO:

1.

4. 3. The pharmaceutical composition of claim 2, formulated for administration by a parenteral or mucosal route.

5. A pharmaceutical composition for treating or preventing an infection caused by a virus that infects epithelial cells of the mammalian respiratory system, comprising a peptide having the amino acid sequence identified as SEQ ID NO: 1 and a pharmaceutically acceptable excipient.

6. 6. The pharmaceutical composition of claim 5, formulated for administration by a parenteral or mucosal route.

7. 6. The pharmaceutical composition of claim 5, wherein the infectious disease is caused by a virus of the Coronaviridae, Orthomyxoviridae and Paramyxoviridae, Picornaviridae, and Adenoviridae families.

8. 6. The pharmaceutical composition of claim 5, wherein the infectious disease is caused by a virus from the group consisting of coronavirus, influenza virus, parainfluenza, respiratory syncytial virus, adenovirus, dengue virus, herpes simplex virus, cytomegalovirus, and swine fever virus.

9. Use of a peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 20 for the manufacture of a medicament for treating or preventing an infection caused by a virus that infects epithelial cells of the mammalian respiratory system.

10. 10. The use according to claim 9, wherein the infectious disease is caused by a virus from the Coronaviridae, Orthomyxoviridae and Paramyxoviridae, Picornaviridae, and Adenoviridae families.

11. 10. The use according to claim 9, wherein the infectious disease is caused by a virus selected from the group consisting of coronavirus, influenza virus, parainfluenza virus, respiratory syncytial virus, adenovirus, dengue virus, herpes simplex virus, cytomegalovirus, and swine fever virus.

12. 10. The use according to claim 9, wherein the medicament comprises two or more peptides having amino acid sequences selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO:

20.

13. A method for treating or preventing an infection caused by a virus that infects epithelial cells of the mammalian respiratory system, comprising administering to an individual in need thereof a therapeutically effective amount of at least one peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO:

20.

14. 14. The method of claim 13, wherein the infectious disease is caused by a virus from the Coronaviridae, Orthomyxoviridae and Paramyxoviridae, Picornaviridae, and Adenoviridae families.

15. 14. The method of claim 13, wherein the infectious disease is caused by a virus selected from the group consisting of coronavirus, influenza virus, parainfluenza virus, respiratory syncytial virus, adenovirus, dengue virus, herpes simplex virus, cytomegalovirus, and swine fever virus.

16. 14. The method of claim 13, wherein an antiviral drug is further administered to the individual.

17. 17. The method of claim 16, wherein the peptide and the antiviral agent are administered simultaneously or sequentially.

18. 17. The method of claim 16, wherein the antiviral agent is selected from the group consisting of ribavirin, ivermectin, penciclovir, nitazoxanide, nafamostat, remdesivir, favipiravir, the peptide identified as SEQ ID NO: 23, alpha interferon (IFN), gamma IFN, or a combination thereof.

19. A combination drug comprising two or more peptides having amino acid sequences selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO:

20.

20. 20. The pharmaceutical combination of claim 19, wherein the peptides are administered simultaneously or sequentially during the same course of treatment.

21. A pharmaceutical combination for treating or preventing an infectious disease caused by a virus that infects epithelial cells of the mammalian respiratory system, comprising: a) at least one peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 20; and b) an antiviral drug.

22. 22. The pharmaceutical combination of claim 21, wherein the antiviral agent is selected from the group consisting of ribavirin, ivermectin, penciclovir, nitazoxanide, nafamostat, remdesivir, favipiravir, the peptide identified as SEQ ID NO: 23, alpha interferon (IFN), gamma IFN, or a combination thereof.

23. 22. The pharmaceutical combination of claim 21, wherein the peptide and the antiviral agent are administered simultaneously or sequentially during the same course of treatment.

24. 22. The pharmaceutical combination of claim 21, wherein the infectious disease is caused by a virus from the Coronaviridae, Orthomyxoviridae and Paramyxoviridae, Picornaviridae, and Adenoviridae families.

25. 22. The pharmaceutical combination of claim 21, wherein the infectious disease is caused by a virus selected from the group consisting of coronavirus, influenza virus, parainfluenza, respiratory syncytial virus, adenovirus, dengue virus, herpes simplex virus, cytomegalovirus, and swine fever virus.

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