Methods and compositions for treating or preventing coronavirus infections

Abstract

To provide a method for treating and / or preventing an infection of Coronavirus.SOLUTION: The method comprises a step of providing a therapeutically effective amount of a composition comprising interferon to a subject via sublingual administration and / or buccal administration, wherein the Coronavirus comprises SARS-CoV-2.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to methods and compositions for treating or preventing coronavirus infection. [Background technology]

[0002] Coronavirus disease 2019 (COVID-19) is an infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 virus). COVID-19 affects different people differently. Most infected individuals develop mild to moderate illness and recover without hospitalization. However, in severe cases, patients can rapidly progress to acute respiratory distress syndrome (SARS), acute fatal lung failure, and / or multiple organ dysfunction syndrome with multiple complications. Nearly all patients have different degrees of lung damage.

[0003] COVID-19 infection induces a sustained release of pro-inflammatory cytokines by inhibiting the secretion of autologous interferon in lung cells, resulting in persistent accumulation of immune cells in the lungs, causing severe inflammation (i.e., cytokine storm) and even necrosis. Most current therapies use viral suppression or immunosuppression as the primary mechanism of treatment.

[0004] Interferons (IFNs) are proteins made by host cells in response to the presence of pathogens such as viruses, bacteria, parasites, or tumor cells. IFNs allow for cell-to-cell communication to trigger the immune system's protective defenses that eradicate the pathogen or tumor. Of the nine different families of human interferons that have been identified, interferon-alpha (IFNα) has been the most widely studied. The U.S. FDA has approved the use of IFNα in large doses given by injection for the treatment of some cancers, as well as hepatitis B and C. In the relevant art of interferons, conventional wisdom is to use high doses of interferon, for example more than 1 million IU, typically administered by intramuscular injection to treat the condition systemically.

[0005] There remains a need for methods and / or compositions to treat and / or prevent COVID-19. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Shackelford et al. (Toxicologic Pathology, Vol 30, No 1, pp93-96, 2002) Summary of the Invention

[0007] The present application describes a method for treating and / or preventing a coronavirus infection, the method comprising: providing a subject with a composition comprising a therapeutically effective amount of interferon via sublingual and / or buccal administration; the coronavirus includes SARS-CoV-2.

[0008] The present application also provides a composition for the treatment and / or prevention of coronavirus infection, the composition comprising a therapeutically effective amount of interferon alpha (INF-α) in the form of a lozenge, the composition being capable of being administered sublingually to a patient. [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 shows changes in body weight in different animal experiments of the present application. [Diagram 2] FIG. 1 shows changes in body weight in different animal experiments of the present application. [Diagram 3] FIG. 1 shows changes in body weight in different animal experiments of the present application. [Figure 4] FIG. 1 shows changes in body weight in different animal experiments of the present application. [Diagram 5] FIG. 1 shows changes in body weight in different animal experiments of the present application. [Figure 6]FIG. 1 shows changes in body weight in different animal experiments of the present application. [Figure 7] Figure showing viral titration of SARS-CoV-2 delta variant on nasal turbinates. [Figure 8] Figure showing viral titer determination of SARS-CoV-2 delta variant on lungs. [Figure 9] FIG. 1 shows viral titration of SARS-CoV-2 omicron variant on nasal turbinates. [Figure 10] FIG. 1 shows viral titration of SARS-CoV-2 Omicron variant in the lungs. [Figure 11A] This figure shows a table of histopathological findings for the placebo group in an animal experiment using the SARS-CoV-2 delta variant. [Figure 11B] This figure shows a table of histopathological findings for the Veldona group in an animal experiment with the SARS-CoV-2 delta mutant strain. [Figure 12A] This figure shows a table of histopathological findings from an animal experiment with the SARS-CoV-2 delta mutant strain. [Figure 12B] This figure shows a table of histopathological findings from an animal experiment with the SARS-CoV-2 delta mutant strain. [Figure 12C] This figure shows a table of histopathological findings from an animal experiment with the SARS-CoV-2 delta mutant strain. [Figure 12D] This figure shows a table of histopathological findings from an animal experiment with the SARS-CoV-2 delta mutant strain. [Figure 13-1] This figure shows a photo of a pathology test in an animal experiment using the SARS-CoV-2 delta mutant strain. [Figure 13-2] This figure shows a photo of a pathology test in an animal experiment using the SARS-CoV-2 delta mutant strain. [Figure 13-3] This figure shows a photo of a pathology test in an animal experiment using the SARS-CoV-2 delta mutant strain. [Figure 14-1]This figure shows a photo of a pathology test in an animal experiment using the SARS-CoV-2 delta mutant strain. [Figure 14-2] This figure shows a photo of a pathology test in an animal experiment using the SARS-CoV-2 delta mutant strain. [Figure 14-3] This figure shows a photo of a pathology test in an animal experiment using the SARS-CoV-2 delta mutant strain. [Figure 15A] This is a table showing histopathological findings in an animal experiment with the SARS-CoV-2 omicron variant. [Figure 15B] This is a table showing histopathological findings in an animal experiment with the SARS-CoV-2 omicron variant. [Figure 15C] This is a table showing histopathological findings in an animal experiment with the SARS-CoV-2 omicron variant. [Figure 15D] This is a table showing histopathological findings in an animal experiment with the SARS-CoV-2 omicron variant. [Figure 16-1] This figure shows a photo of a pathology examination in an animal experiment using the SARS-CoV-2 omicron mutant strain. [Figure 16-2] This figure shows a photo of a pathology examination in an animal experiment using the SARS-CoV-2 omicron mutant strain. [Figure 16-3] This figure shows a photo of a pathology examination in an animal experiment using the SARS-CoV-2 omicron mutant strain. [Figure 16-4] This figure shows a photo of a pathology examination in an animal experiment using the SARS-CoV-2 omicron mutant strain. [Figure 17] This figure shows a photo of a pathology examination in an animal experiment using the SARS-CoV-2 omicron mutant strain. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] In one aspect, the present application provides a method for treating and / or preventing a coronavirus infection, the method comprising: providing to a subject via sublingual and / or buccal administration a composition comprising a therapeutically effective amount of interferon; the coronavirus includes SARS-CoV-2.

[0011] The coronavirus may be Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), SARS-CoV-1, Middle East Respiratory Syndrome-associated Coronavirus (MERS-CoV), etc. In one embodiment, the coronavirus is SARS-CoV-2. Clinically, SARS-CoV-2 has several mutant strains, such as Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Delta (B.1.617.2), Mu (B.1.621), R.1, Epsilon, Theta, and Zeta. The treatment and / or prevention method of the present invention can be applied to all mutant strains of SARS-CoV-2.

[0012] The method of the present application is characterized by sublingual and / or buccal administration of a composition comprising an interferon. Examples of suitable interferons include type I INF, type II INF, and type III INF.

[0013] In a preferred embodiment, the composition comprises interferon alpha (INF-α). The INF-α may be natural INF-α, recombinant INF-α, or a mixture thereof. In a preferred embodiment, the INF-α contained is recombinant INF-α.

[0014] In some embodiments, the composition may further comprise interferon beta and / or interferon gamma.

[0015] In contrast to conventional wisdom, the present application provides compositions having low doses of IFNα for oral delivery that are a non-toxic alternative to high doses of injectable IFNα. More specifically, the low doses of IFNα are given via sublingual and / or buccal administration.

[0016] Sublingual administration involves placing the composition under the tongue and allowing it to dissolve and be absorbed into the subject's blood via the tissues under the tongue. Buccal administration involves placing the composition between the gums and cheek where it dissolves and is absorbed into the subject's blood. Generally, sublingually and / or buccally administered compositions pass through and are not absorbed by the digestive tract.

[0017] Examples of dosage forms for sublingual and / or buccal administration include, but are not limited to, lozenges, tablets, films, and sprays. In one embodiment, the composition is in the form of a lozenge. In one embodiment, the composition is in the form of an orally disintegrating tablet. Orally disintegrating tablets can disintegrate in the oral cavity within seconds without the need for additional liquid.

[0018] Thus, via sublingual and / or buccal administration, the oral mucosa can rapidly contact and absorb the composition, and the therapeutic effect can be obtained more efficiently. In addition, sublingual and / or buccal administration can develop higher drug compliance.

[0019] In one embodiment, the composition comprises an IFN-α dose of 1,000 International Units (IU) or less. In one embodiment, the composition comprises an IFN-α dose of 1,000 micrograms or less. The dosage can be, for example, less than 900 IU, less than 800 IU, less than 700 IU, less than 600 IU, less than 500 IU, less than 400 IU, less than 300 IU, less than 200 IU, or less than 100 IU.

[0020] In one embodiment, the composition comprises a dose of INF-α of 1 IU or more, for example, greater than 2 IU, greater than 3 IU, greater than 4 IU, greater than 5 IU, greater than 10 IU, greater than 25 IU, greater than 50 IU, greater than 75 IU, greater than 80 IU, or greater than 90 IU.

[0021] In one embodiment, the composition comprises a dose of INF-α between 1 IU and 1,000 IU, e.g., 2 IU, 3 IU, 4 IU, 5 IU, 8 IU, 10 IU, 25 IU, 50 IU, 75 IU, 80 IU, 90 IU, 100 IU, 200 IU, 300 IU, 400 IU, 500 IU, 600 IU, 700 IU, 800 IU, 900 IU, 950 IU, or a value within a range between any two of the above values.

[0022] In a preferred embodiment, the amount of INF-α contained in the composition may be 1 IU to 500 IU. In another preferred embodiment, the amount of INF-α contained in the composition may be 1 IU to 100 IU. In another preferred embodiment, the amount of INF-α contained in the composition may be 5 IU to 50 IU. In a more preferred embodiment, the amount of INF-α contained in the composition may be 1 IU to 10 IU.

[0023] The composition may include a buffer, carrier, excipient, or the like.

[0024] Examples of suitable buffers include, but are not limited to, acetate buffers, phosphate buffers, citrate buffers, borate buffers, etc. In some embodiments, the buffer may be phosphate buffered saline (PBS), saline, etc.

[0025] Examples of suitable carriers can include, but are not limited to, carbohydrates, antioxidants, chelating agents, low molecular weight proteins, etc. In some embodiments, the carrier can be glucose, sucrose, dextran, ascorbic acid, glutathione, etc.

[0026] Examples of suitable excipients may include, but are not limited to, diluents, coating agents, colorants, lubricants, preservatives, flavoring agents, etc. In some embodiments, the excipient may be ethanol, lactose, starch, etc.

[0027] In one embodiment, the method for treating and / or preventing coronavirus infection further comprises: administering to the subject a therapeutically effective amount of an antiviral agent, an anti-inflammatory agent, interferon beta, and / or interferon gamma. Examples of antiviral agents include, but are not limited to, remdesivir, paquilovid, molnupiravir, and zocova. Such administration may occur prior to, simultaneously with, or after administration of the composition comprising INF-α. The route of such administration is not limited, and for example, the antiviral agent may be administered orally or via injection or inhalation.

[0028] In another aspect, the present application also provides a composition for the treatment and / or prevention of coronavirus infection, comprising a therapeutically effective amount of interferon alpha (INF-α) in a sublingual and / or buccal administration dosage form.

[0029] In one embodiment, the INF-α is recombinant INF-α.

[0030] In one embodiment, INF-α is between 1 IU and 1,000 IU, for example, 2 IU, 3 IU, 4 IU, 5 IU, 8 IU, 10 IU, 25 IU, 50 IU, 75 IU, 80 IU, 90 IU, 100 IU, 200 IU, 300 IU, 400 IU, 500 IU, 600 IU, 700 IU, 800 IU, 900 IU, 950 IU, or a value within a range between any two of the above values.

[0031] In one embodiment, the composition further comprises a buffer, a carrier, and / or an excipient. In some embodiments, the buffer may be phosphate buffered saline (PBS).

[0032] In one embodiment, the composition further comprises interferon beta and / or interferon gamma.

[0033] In one embodiment, the composition further comprises an antiviral agent and / or an anti-inflammatory agent.

[0034] In the present application, dosage forms for sublingual and / or buccal administration may include lozenges, tablets, films, sprays, etc. In one embodiment, the composition is in the form of a lozenge.

[0035] In the present application, the composition is delivered intraorally as a lozenge containing low doses of IFN-α, i.e., 1-1000 IU. Orally administered IFNα can activate dozens of immune system genes in peripheral blood and can be effective against viral diseases without the side effects associated with injections of high doses of IFNα.

[0036] In the present application, sublingual and / or buccal administration of low doses of IFNα is rapidly absorbed by the subject, prevents digestion-induced loss of therapeutic efficacy, and may promote greater compliance.

[0037] Working Example The Golden Syrian hamster model is recommended by BARDA (Biomedical Advanced Research and Development Authority, Health and Human Services, US) as an indicator of in vivo efficacy since it can be infected without artificial genetic manipulation. In practice, hamsters recover from infection in 7-10 days, and the common clinical sign is weight loss (10-20%) without fever. In this application, drug efficacy for virus eradication will be evaluated through primary indicators of viral load in the nose and lungs. Also, the rate of weight change during infection will be evaluated. Secondary indicators are also proposed to evaluate the plasma levels of several key cytokines using qPCR to investigate possible drug mechanisms of action.

[0038] In the following examples, male golden hamsters were obtained from the National Laboratory Animal Center (National Applied Research Laboratories, Taiwan). All experiments were performed in an ABSL-3 core facility (Institute of Preventive Medicine, National Defense Medical College). Hamsters were randomly assigned to experimental groups from different litters and were acclimated in the ABSL-3 facility for one week prior to the experiment. The study protocol was reviewed and approved by the Committee on the Institutional Animal Care and Use, Institute of Preventive Medicine (Permit number: AN-111-11). EXAMPLES

[0039] Materials and Methods Cell lines and viruses Vero E6 cells were cultured in DMEM medium with 10% fetal bovine serum. In all cases, the passage number of the cells was less than 25. SARS-CoV-2 virus was provided by the Taiwan Center for Disease Control (hCoV-19 / Taiwan / 1144 / 2021; B.1.617.2; EPI_ISL_5854263) and propagated in Vero E6 cells. SARS-CoV-2 titers were determined via plaque assay and processed in a BSL-3 or BSL-4 facility.

[0040] SARS-CoV-2 delta and omicron variants were applied in this application.

[0041] Animal testing Twelve hamsters (~5 to 6 weeks old) were exposed to SARS-CoV-2 (delta variant) via intranasal administration (50ul / nare) on day 0 (dose: 1 × 10 4pfu / ml). The hamsters were then divided into two groups, one of which was administered the composition of the present application (hereinafter sometimes simply referred to as the "Veldona group"), and the other was administered a buffer solution as a placebo group (hereinafter also referred to as the "control group"). In this example, the composition of the present application has a dose of 8 IU / 100 g (QD) of interferon alpha. No toxicity was observed at this dose for hamsters. The hamsters were sacrificed for the following analysis.

[0042] For testing SARS-CoV-2 delta variants, hamsters were administered the compositions of the present application daily, beginning 6 hours after viral infection on day 0. Hamsters were sacrificed on days 3 and 6.

[0043] For testing the SARS-CoV-2 Omicron variant, hamsters were dosed prior to virus challenge for 5 days (days -5 to -1) and daily starting 6 hours after virus infection on day 0. The total treatment period was 16 days. Hamsters were sacrificed on days 2, 5, and 10.

[0044] Weight change Hamsters were weighed daily throughout the study and changes in body weight were calculated.

[0045] Lung and nasal turbinate processing steps The lung tissues were transferred to 2 mL tubes containing 1 mL of DMEM medium and 3 mm glass beads, respectively, and pulverized using a Tissue Lyser machine (Bertin / Precellys24). 100 μL of the supernatant medium (for lung RT-qPCR) was transferred to a 2 mL tube containing 0.9 mL of TRIzol reagent for RNA extraction. 100 μL of the supernatant medium was ready to perform the plaque assay. For backup, 250 μL of both supernatant mediums were stored at -80 °C. Extraction of the supernatant for nasal turbinate RT-qPCR followed the procedure for lung tissue processing.

[0046] Quantitative real-time RT-PCR (RT-qPCR) assay 100 μL of tissue supernatant was taken for RNA extraction via TANBead Nucleic Acid Extraction Kit. After RNA quantification, 1 μg of RNA was reverse transcribed into cDNA using SuperScript IV Reverse Transcriptase Kit (Invitrogen). RT-qPCR was then performed using PowerTrack SYBR Green Master Mix (Applied biosystems) and LightCycler480 system using primer pairs specific for the NP gene. Target genes for RT-qPCR included SARS-CoV-2 NP gene, IFN-α(-2b), IFN-β, IFN-λ, IL-6, TNF-α, and TGF-β, and GAPDH was included as an internal control.

[0047] Pathological Sectioning and Interpretation of Lung Tissue The National Laboratory Animal Center was commissioned to perform HE staining and interpretation of pathological sections of lung tissue. Histopathological evaluation was performed on the submitted lungs. The severity of the lesions (excluding inflammatory areas of the lungs) was graded according to the method described by . The degree of lesions was histopathologically graded from 0 to 5 according to severity (0=absent; 1=minimal (<1%); 2=slight (1-25%); 3=moderate (26-50%); 4=moderate (51-75%); 5=severe / high (76-100%)).

[0048] Lesions (from the left lobe) will be visually assessed at necropsy and photographs of the lungs will be taken as evidence of any gross findings. Tissue (from the left lobe) will be collected in cassettes for histopathological H&E staining.

[0049] result Several independent animal experiments were performed and the results are shown as follows. Figures 1 to 6 show the percentage of body weight change in different animal experiments. Figures 1 to 3 were for the SARS-CoV-2 delta mutant strain, and Figures 4 to 6 were for the SARS-CoV-2 omicron mutant strain. As shown in the figures, the hamsters were most severely affected during the first three days after infection with the virus, and their body weight rapidly decreased. About 7 to 10 days after infection, the hamsters recovered and regained their weight. Compared with the control group, the results showed that the hamsters in the Veldona group regained significant weight on the fourth day, almost returning to their pre-infection state. Then, the body weight remained stable. This indicates that the composition of the present application has a significant anti-COVID-19 virus effect.

[0050] Figures 7 and 8 show viral titration of SARS-CoV-2 delta variants on nasal turbinates and lungs, respectively. Figures 9 and 10 show viral titration of SARS-CoV-2 omicron variants on nasal turbinates and lungs, respectively.

[0051] According to the results shown in Figures 7 and 8, the amount of SARS-CoV-2 delta mutant strain in the nasal cavity of the Veldona group on day 3 after infection was similar to that of the control group, but the amount of virus in four hamsters was significantly reduced on day 6 after infection. More importantly, at the site of infection in the lower respiratory tract, the Veldona group had less virus in the lungs of hamsters on day 3 after infection than the placebo group. On day 6 after infection, the hamsters had a significantly reduced amount of virus.

[0052] According to the results shown in Figures 9 and 10, the Veldona group had significantly less virus in the lungs of hamsters than the placebo group on day 5 after infection, and there was no inflammation on day 10. It was found that on day 10 after infection, the hamsters in the Veldona group had a significantly reduced amount of virus in their lungs. Combining the above two results, the hamsters in the Veldona group show good anti-COVID-19 efficacy during the 15-day treatment period.

[0053] Figures 11A and 11B show histopathological findings for the placebo group and the Veldona group, respectively, in one animal experiment with the SARS-CoV-2 delta variant. Figures 12A to 12D show histopathological findings in another animal experiment with the SARS-CoV-2 delta variant.

[0054] Figures 13 and 14 show the photos of pathology examination in one animal study of SARS-CoV-2 delta mutant strain. The left column of Figure 13 shows lung tissue sections with H&E staining, indicating multifocal pulmonary inflammation in the lung lobes by arrows, and further indicating artificial damage (extravasated blood cells in the alveolar lumen) or artifacts related to the autopsy procedure by circled areas. The right column of Figure 13 shows a partial enlargement of the lung section, i.e., a high magnification (200X) of the boxed area shown in the left column. The animal ID and observed symptoms were noted under each photograph. The animal ID and its detailed information are shown in Table 1.

[0055] [Table 1] In one of the animal experiments with SARS-CoV-2 delta variant, the results of histopathological examination included the microscopic changes presented in the histopathological findings table in Figures 12A-12D, and the pathological photographs presented in Figures 13-14. Histologically, Syrian hamsters infected with SARS-CoV-2 (delta variant) presented lesions including:

[0056] Mixed cell type inflammation, peribronchial infiltrate, and perivascular infiltrate: At 3 dpi (days post inoculation), lesions were characterized by a mixture of lymphoid and histiocytic cell types and heterophylls within the alveoli / interstitium, and peribronchiolar and perivascular spaces in the lungs. Lesion severity was minimal to slight. At 6 dpi, hamsters developed slight to moderate interstitial inflammation of the lungs with mixed cell types, primarily mononuclear cells with little heterophylls.

[0057] Degeneration / necrosis of bronchial epithelial cells with or without inflammatory infiltrates within the bronchioles: Lesions were observed in the lungs of animals inoculated with SARS-CoV-2 and were characterized by pyknosis, cellular swelling, cytoplasmic vacuolization, and perinuclear clear spaces in the affected epithelium together with heterophilic infiltration into the bronchial lumen. Lesions were of minimal to moderate severity.

[0058] Regenerative hyperplasia of the bronchiolar epithelium: Lesions were primarily observed in the lungs of animals inoculated with SARS-CoV-2 at 6 dpi. Epithelial regenerative hyperplasia is a common response to epithelial injury. Bronchiolar epithelial hyperplasia is characterized by an increased layer of superficial respiratory epithelial cells that normally lack cilia. Lesions were minimal in severity.

[0059] Alveolar wall necrosis: Lesions were observed in the lungs of animals inoculated with SARS-CoV-2 and were characterized by cellular swelling and pyknosis of nuclei in the affected epithelium. The severity of the lesions was minimal to slight.

[0060] Type II alveolar cell hyperplasia: Lesions were primarily observed in the lungs of animals inoculated with SARS-CoV-2 at 6 dpi. Hyperplasia of type II pneumocytes was characterized by reactive hypertrophy with megakaryons, giant cells, and scattered syncytia. Lesions were minimally severe.

[0061] Vasculitis and endothelialitis: Lesions were observed in the lungs of animals inoculated with SARS-CoV-2. Endotheliitis is characterized by bulging of blood vessels into the lumen with infiltration of macrophages and lymphocytes. Inflammation of blood vessels (vasculitis) is characterized by degeneration of fibrinoids and inflammatory infiltration of heterophylls and lymphocytes. Lesions were minimal to moderate in severity.

[0062] Pulmonary parenchymal edema and hemorrhage: Pulmonary hemorrhage is characterized by the accumulation of extravasated blood cells in the alveolar lumen. Pulmonary edema is characterized by the accumulation of homogeneous eosinophilic material in the alveolar lumen. Lesions were observed in some lung submissions. Hemorrhage and edema result from either alterations in pulmonary circulation or damage to the air-blood barrier in the alveolar wall. The severity of the lesions was minimal to subtle. In addition, pleural thickening was observed in some lung submissions. The lesions may be due to SARS-CoV-2 inoculation. Alveolar calcification was observed in some lung submissions. The lesions may be due to spontaneous or incidental lesions. Extravasated blood cells in the alveolar lumen associated with artificial injury were thought to be related to injury from tissue sampling.

[0063] The pathological indicators in which the Veldona group was superior to the control group were all lung symptoms of COVID-19. For example, in the Veldona group: (1) no lung under-inflation was observed at 6 dpi, which is a condition associated with pneumonia and may lead to acute respiratory distress syndrome (ARDS), shortness of breath, fatigue, difficulty in inhalation, and exercise intolerance; (2) a lower degree of vasculitis and endotheliitis, which may lead to thrombosis; (3) less hyperplasia of type II alveolar epithelial cells could be observed. Lung under-inflation can lead to pulmonary fibrosis and ARDS. Patients with ARDS usually require intensive care and can cause a mortality rate of 40-70%.

[0064] In this study, although the severity of pneumonia in the left lobe of the lung was not significantly different between the Veldona group and the control group at 3 dpi and 6 dpi, the Veldona group had lower expression of IL-6 and TNF-alpha, and higher expression of IFN-alpha, IFN-gamma, and IFN-lambda, which were confirmed by the results of cytokine qPCR in lung tissue. Obviously, the Veldona group had a better antiviral immune response without having a cytokine storm. According to the above results, the Veldona group with moderate pneumonia (40-50%) had a lower viral load compared to the control group.

[0065] Figures 15A-15D show histopathological findings in one animal experiment with SARS-CoV-2 Omicron mutant. Figures 16 and 17 show photos of pathology examination in one animal experiment with SARS-CoV-2 Omicron mutant. The left column of Figure 16 shows lung tissue sections with H&E staining (12.5X), and the right column shows partial enlargement of the lung sections, i.e., high magnification (200X) of the boxed area shown in the left column. The animal sample ID and observed symptoms were noted under each photo. The animal ID and its detailed information are shown in Table 2.

[0066] [Table 2] In one of the animal studies of SARS-CoV-2 Omicron variant, the results of histopathological examination included the microscopic changes presented in the histopathological findings table in Figures 15A-15D, and the pathological photographs presented in Figures 16-17. Histologically, Syrian hamsters infected with SARS-CoV-2 (Omicron variant) presented lesions including:

[0067] Mixed cell type inflammation, peribronchial infiltrate, and perivascular infiltrate: Lesions were characterized by a mixture of lymphoid and histiocytic cell types and heterophylls within the alveolar / interstitium, as well as peribronchial and perivascular lymphoid and histiocytic cell types in the lungs. Lesions were minimal in severity.

[0068] Degeneration / necrosis of bronchial epithelial cells with or without inflammatory infiltrates within the bronchioles: Lesions were observed in the lungs of animals inoculated with SARS-CoV-2 and were characterized by pyknosis, cellular swelling, cytoplasmic vacuolization, and perinuclear zona pellucida in the affected epithelium together with heterophilic infiltration into the bronchial lumen. Lesions were minimal in severity.

[0069] Vasculitis and endotheliitis: Lesions were observed in the lungs of animals inoculated with SARS-CoV-2. Endotheliitis is characterized by bulging of blood vessels into the lumen with infiltration of macrophages and lymphocytes. The severity of the lesions was minimal.

[0070] In addition, bone metaplasia in the alveoli and alveolar calcification were observed in some lungs. The lesions may be due to spontaneous or accidental lesions. Extravasated blood cells in the alveolar lumen associated with artificial injury were thought to be related to injury due to tissue sampling.

[0071] Lesions associated with SARS-CoV-2 inoculation include (1) mixed cell type inflammation, peribronchial infiltrates, and perivascular infiltrates; (2) bronchial epithelial cell degeneration / necrosis with or without inflammatory infiltrates within the bronchioles; and (3) vasculitis and endotheliitis. Pulmonary hypoinflation and alveolar-filled red blood cells (artifactual findings) were observed in most submitted lungs, and histopathological examination was limited by these artifacts.

[0072] This study evaluated the efficacy of the compositions of the present application over a 16-day course of treatment (5 days pre-treatment and 10 days post-infection treatment) of hamsters infected with Omicron mutant strains. Compared to hamsters receiving a solution without the test substance ("control group"), hamsters receiving a solution with the test substance ("Veldona group") demonstrated resistance to weight loss immediately after infection, and then showed a trend toward better recovery over the next 3 days. The weights of hamsters in the Veldona group remained more stable over the treatment period than hamsters in the control group.

[0073] Regarding pathological indices, hamsters in the Veldona group showed no mixed cell type inflammation, peribronchial infiltration, and perivascular infiltration, compared with 50% in the control group, on day 10. Hamsters in the Veldona group generally showed promising results in terms of therapeutic indications for new mutant virus infection.

[0074] Viral infection occurs primarily in the upper respiratory tract, but in severe cases, it spreads to the lower respiratory tract, i.e., the lungs, causing severe and irreversible complications. In this COVID-19 antiviral efficacy study, with reference to the results shown in Figures 1-3, 7, 8, 11A-11B, 12A-12D, 13 and 14, it was found that hamsters infected with the delta mutant strain that received the composition orally once a day maintained and regained weight significantly, similar to the control. After a 7-day course of treatment, the Veldona group has effectively reduced the virus. Combined with the physiological characteristics and pathological findings, the results show that the composition of the present application with a low dose of interferon in an oral / sublingual formulation has a protective effect against the lung organ caused by SARS-CoV-2 by modulating the immune response. It also allows infected animals to recover to a normal physiological state more quickly. The composition of the present application can induce significant systemic immunomodulation, has a good antiviral effect on COVID-19, and can be an effective treatment.

[0075] With reference to the results shown in Figures 4-6, 9, 10, 15A-15D, 16, and 17, it was found that hamsters infected with the Omicron mutant strain receiving the composition of the present application orally once a day maintained significant body weight and recovered similarly to the control. After a 16-day course of treatment, the Veldona group can effectively reduce the virus in this preventive study design. Combined with the physiological characteristics and pathological findings, the results show that the composition of the present application has a protective effect against the lung organ caused by SARS-CoV-2 by modulating the immune response. It also allows infected animals to recover to a normal physiological state more quickly. The composition of the present application can induce significant systemic immunomodulation, has a good antiviral effect on COVID-19, and can be an effective treatment.

[0076] In this application, the composition can activate the autoimmune system to achieve the effect of fighting various viruses, including SARS-COV-2. The composition can induce cytotoxicity and activate NK cells and antibody-dependent cytotoxicity. The composition containing recombinant human interferon against coronavirus has obvious antiviral effect and at the same time has great therapeutic potential in preventing coronavirus infection.

[0077] Although the present invention has been disclosed by referring to the preferred embodiments and examples detailed above, it will be understood that these examples are intended to be illustrative and not limiting. Modifications and combinations will readily occur to those skilled in the art, and it is contemplated that these modifications and combinations are within the spirit of the present invention and the scope of the appended claims and equivalent systems and methods.

Claims

1. 1. Use of a composition comprising an interferon in the manufacture of a medicament for the treatment and / or prevention of a coronavirus infection, said composition being in a dosage form for sublingual and / or buccal administration, said composition comprising an INF-α dose of less than 1,000 IU, and said coronavirus comprising SARS-CoV-2.

2. The use according to claim 1, wherein the INF-α is recombinant INF-α.

3. The use according to claim 1, wherein the composition comprises a dose of 1 IU or more of INF-α.

4. The use of claim 1, wherein the composition comprises a dose of INF-α of from 1 IU to less than 1,000 IU.

5. The use of claim 1 , wherein the composition further comprises a buffer, a carrier, and / or an excipient.

6. The use according to claim 1 , wherein the composition further comprises interferon beta and / or interferon gamma.

7. The use according to claim 1 , wherein the composition further comprises an antiviral agent and / or an anti-inflammatory agent.

8. The use according to claim 1 , wherein the composition is in the form of a lozenge, tablet, film, or spray.

9. A composition for the treatment and / or prevention of coronavirus infection, comprising a therapeutically effective amount of interferon alpha (IFN-α), in a sublingual and / or buccal administration dosage form, wherein the therapeutically effective amount of INF-α is less than 1,000 IU.

10. The composition of claim 9, wherein the INF-α is recombinant INF-α.

11. The composition of claim 9, wherein the INF-α is from 1 IU to less than 1,000 IU.

12. The composition of claim 9 further comprising a buffer, a carrier, and / or an excipient.

13. 10. The composition of claim 9, further comprising interferon beta and / or interferon gamma.

14. 10. The composition of claim 9, further comprising an antiviral agent and / or an anti-inflammatory agent.

15. The composition of claim 9 in the form of a lozenge, tablet, film, or spray.

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