Compositions and methods for treating or preventing diseases associated with beta-coronavirus infection

Abstract

The present disclosure relates to methods and compositions for treating or preventing betacoronavirus infections, including SARS-CoV-2 and COVID-19, or inhibiting symptoms of disease caused by betacoronavirus infection in a subject. In various embodiments, the compositions and methods include a combination of a nonsteroidal anti-inflammatory drug (NSAID) and ketotifen. In exemplary embodiments, naproxen and / or indomethacin are combined with ketotifen for therapy in a single unit dose or in separate compositions, and this combination is surprisingly and unexpectedly effective in reducing SARS-CoV-2 viral load and treating COVID-19.

Description

[Technical Field]

[0001] This disclosure comprises compositions and methods for treating or preventing diseases associated with beta-coronavirus infection. [Background technology]

[0002] Coronaviruses are a large family of viruses that cause illnesses ranging from the common cold to more serious diseases such as Middle East Respiratory Syndrome (MERS-CoV), Severe Acute Respiratory Syndrome (SARS-CoV), and Coronavirus Disease 2019 (COVID-19). Coronaviruses are zoonotic, meaning they are transmitted between animals and humans. COVID-19 is an infectious disease caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The SARS-CoV-2 virus is an enveloped, positive-sense, single-stranded RNA beta-coronavirus. This disease has been spreading globally since 2019.

[0003] SARS-CoV-2 is often spread through respiratory droplets produced during coughing. The time from exposure to symptom onset is generally 2 to 14 days, with an average of 5 days. The standard diagnostic method is reverse transcription polymerase chain reaction (rRT-PCR) from nasopharyngeal swabs or sputum samples. Antibody assays can also be used with serum samples. The infection can also be diagnosed from a combination of symptoms, risk factors, and chest CT scans that exhibit features of pneumonia.

[0004] COVID-19 is a novel, poorly understood disease for which effective treatments are limited. Common symptoms of COVID-19 include fever, cough, and shortness of breath. Muscle aches, phlegm production, and sore throat are some of the less common symptoms. The majority of cases remain mild, but some progress to pneumonia, severe acute respiratory syndrome, multiple organ failure, and death. The mortality rate of cases is estimated to be between 1% and 5%, varying with age and other health conditions. The disease is more likely to occur in people whose immune systems are compromised due to age, immunosuppressive therapy, chronic illness, psychological stress, or other factors.

[0005] Therefore, there is a need for effective and safe methods to prevent or treat infections caused by beta-coronaviruses such as SARS-CoV-2, and / or to alleviate the symptoms of related diseases (e.g., COVID-19). [Overview of the project]

[0006] This disclosure relates to methods and compositions for treating or preventing betacoronavirus infections, including SARS-CoV-2 and COVID-19, or for alleviating symptoms of diseases caused by betacoronavirus infections in subjects. In various embodiments, compositions and methods include a combination of a nonsteroidal anti-inflammatory drug (NSAID) and ketotifen. In exemplary embodiments, naproxen and / or indomethacin are combined with ketotifen for therapy in a single unit dose or in separate compositions, and this combination is remarkably and unexpectedly effective in reducing the SARS-CoV-2 viral load and / or alleviating COVID-19 symptoms.

[0007] In various embodiments, the method comprises administering one or more pharmaceutical compositions to a mammal, typically a human, in an amount effective to treat or prevent a disease caused by SARS-CoV-2 and / or to alleviate its symptoms. An example of a disease caused by SARS-CoV-2 is coronavirus disease 2019 (i.e., COVID-19).

[0008] In some embodiments, the disclosure provides a treatment for subjects at risk of developing disease caused by SARS-CoV-2 (or other beta-coronaviruses). In some embodiments, subjects are at risk of developing disease caused by SARS-CoV-2 due to age, immunodeficiency, or one or more pre-existing conditions.

[0009] In various embodiments, therapy may be initiated before or after the onset of COVID-19 symptoms. In some embodiments, therapy is initiated shortly after the first appearance of symptoms (i.e., immediately after the first appearance of symptoms). In some embodiments, subjects according to the Disclosure are asymptomatic but test positive for SARS-CoV-2 (or other beta-coronavirus). In some embodiments, therapy according to the Disclosure is initiated as soon as the individual is first suspected of having an infection or disease caused by SARS-CoV-2 (or other beta-coronavirus), but before the onset of any symptoms of the disease. In various embodiments, subjects have one or more risk factors for COVID-19. In various embodiments, subjects are experiencing one or more symptoms selected from: throat / nasal congestion, cough, fever, loss of taste, loss of smell, muscle pain, arthralgia (myalgia), fatigue, sore throat, headache, myocarditis, coagulation disorder, kidney injury or failure, liver injury or failure, neurological disorder, hypoxia, pneumonia with or without hypoxia, acute respiratory syndrome, or multiple organ failure.

[0010] In various embodiments, the present invention provides a method for administering one or more pharmaceutical compositions to a human subject in an amount effective to treat or prevent beta-coronavirus infection or related disease in the subject. The NSAID and ketotifen may be administered as a single unit dose (composition) or in separate compositions. In some embodiments, the method of the present disclosure includes administering an effective amount of indomethacin and ketotifen. In certain embodiments, indomethacin and ketotifen are administered at least once daily, with a daily dose of indomethacin being about 20 mg to about 300 mg, about 40 mg to about 200 mg, or about 75 mg to about 150 mg. In various embodiments, the daily dose of ketotifen is about 1 mg to about 10 mg, or about 1 mg to about 6 mg, or about 2 mg to about 5 mg, or about 3 mg to about 4 mg. In some embodiments, the method of the present disclosure includes administering naproxen and ketotifen. In certain embodiments, naproxen and ketotifen are administered at least once daily, with daily doses of naproxen ranging from approximately 100 mg to approximately 1600 mg, approximately 200 mg to approximately 1400 mg, approximately 400 mg to approximately 1200 mg, approximately 600 mg to approximately 1000 mg, or approximately 700 mg to approximately 900 mg. In various embodiments, daily doses of ketotifen are approximately 1 mg to approximately 10 mg, or approximately 1 mg to approximately 6 mg, or approximately 2 mg to approximately 5 mg, or approximately 3 mg to approximately 4 mg. In certain embodiments, the unit dose of this disclosure comprises 50 mg of indomethacin and 1 mg of ketotifen, and this unit dose is administered two or three times daily.

[0011] In some embodiments, the disclosure provides pharmaceutical compositions comprising an NSAID (e.g., indomethacin and / or naproxen) and ketotifen as active pharmaceutical ingredients. In some embodiments, the pharmaceutical composition comprises an NSAID (e.g., indomethacin and / or naproxen) and ketotifen as active pharmaceutical ingredients. In these embodiments, the composition may include inactive ingredients, such as pharmaceutical excipients, sweeteners, flavorings, taste masking ingredients, diluents, alum, stabilizers, buffers, colorants, respiratory neutralizers (e.g., peppermint or menthol scents), emulsifiers, suspending agents, and agents that control delivery. The compositions in some embodiments may have additional ingredients (including the active ingredients) that do not affect the basic and novel properties of the composition.

[0012] According to embodiments of the present disclosure, the pharmaceutical composition(s) may be administered orally, intravenously, intramuscularly, subcutaneously, transdermally, by inhalation, or intranasally. In further embodiments, the composition comprising ketotifen and an NSAID, such as indomethacin and / or naproxen, may be combined in the form of tablets, capsules, lozenges, or chewing gum.

[0013] Other aspects and embodiments of this disclosure will become apparent from the following detailed description. [Brief explanation of the drawing]

[0014] [Figure 1] This illustrates the progression of COVID-19 disease: the early infection phase with activation of the innate immune system, the pulmonary phase with activation of the adaptive immune system, and the hyperinflammatory phase with cytokine storm. [Figure 2] The dose-response curves for naproxen and ketotifen, both individually and in combination, are shown. EC50 represents the effective concentration of the product, i.e., the concentration at which viral infection is inhibited by 50 percent. [Figure 3] The dose-response curves for indomethacin and ketotifen, both individually and in combination, are shown.

Mode for Carrying Out the Invention

[0015] The present disclosure relates to methods and compositions for treating or preventing beta-coronavirus infections, including SARS-CoV-2 and COVID-19, or for reducing the symptoms of diseases caused by beta-coronavirus infections in a subject. In various embodiments, the compositions and methods include a combination of a non-steroidal anti-inflammatory drug (NSAID) and ketotifen. In an exemplary embodiment, naproxen and / or indomethacin are combined with ketotifen for therapy in a single unit dose or in separate compositions, and this combination is surprisingly and unexpectedly effective in reducing the SARS-CoV-2 viral load and / or reducing COVID-19 symptoms.

[0016] Thus, the present disclosure provides therapeutic and prophylactic pharmaceutical compositions for diseases caused by beta-coronaviruses, including SARS-CoV-2. The SARS-CoV-2 virus is an enveloped, positive-sense single-stranded RNA beta-coronavirus. In embodiments, the methods and compositions of the present disclosure are effective in treating, reducing, or preventing the symptoms of diseases caused by distinguishable genetic variants of SARS-CoV-2, including the B.1.1.7 (i.e., alpha), B.1.351 (i.e., beta), p.1 (i.e., gamma), B.1.617.2 (i.e., delta), and C.37 (i.e., lambda) variants of SARS-CoV-2.

[0017] In various embodiments, the method involves administering one or more pharmaceutical compositions to a mammal, typically a human, in an amount effective to treat or prevent a disease caused by SARS-CoV-2 and / or to alleviate its symptoms. An example of a disease caused by SARS-CoV-2 is coronavirus disease 2019 (i.e., COVID-19). The clinical progression of COVID-19 ranges from asymptomatic carrierhood to a severe cytokine storm with respiratory failure, multi-organ dysfunction, and ultimately death. Early infection (or Stage I) is characterized by activation of the innate immune system and presents with increased cytokine and chemokine production. This stage is the incubation period during which SARS-CoV-2 reproduces and establishes residence in the host, primarily focusing on the respiratory system. Symptoms reported at the time of early infection are usually mild and often nonspecific, and include fever, cough, diarrhea, and headache. COVID-19 infection progresses to the pulmonary stage (or "stage II"), which can be divided into two distinct parts: pneumonia without hypoxia (stage IIA) or pneumonia with hypoxia (stage IIB). In stage IIB, symptoms worsen, and patients are likely to require hospitalization and oxygen support. Some infections progress to stage III, which manifests as extrapulmonary systemic hyperinflammatory syndrome caused by a cytokine storm. Stage III is characterized by fervent inflammation due to high levels of circulating cytokines such as interleukin (IL)-6 and tumor necrosis factor alpha (TNF-α). These inflammatory mediators cause acute respiratory distress syndrome (ARDS) and multi-organ damage, including cardiac and renal dysfunction. Generally, the prognosis and recovery from stage III are poor.

[0018] Mast cells are strongly activated during SARS-CoV-2 infection, causing damage to lung tissue, resulting in not only bleeding in the lungs and accumulation of fluid in the lungs, but also causing other COVID-19 symptoms. Traditionally, mast cell stabilizers such as ketotifen have been used in humans for the treatment of allergies and asthma, and these drugs are thought to act by preventing mast cell degranulation. Thus, in various embodiments, the present invention involves a combination of a mast cell stabilizer and an NSAID for preventing or treating coronavirus infection or related diseases. An example of a mast cell stabilizer is ketotifen.

[0019] In embodiments, the present disclosure provides treatment for a subject at risk of developing a disease caused by SARS-CoV-2 (or other betacoronaviruses). As used herein, a subject at risk of developing a disease caused by SARS-CoV-2 is either not infected with SARS-CoV-2 or other betacoronaviruses (i.e., negative for SARS-CoV-2 testing), or otherwise not known to be infected with SARS-CoV-2. In some embodiments, a subject at risk of developing a disease caused by SARS-CoV-2 has had close contact with one or more SARS-CoV-2 positive individuals. As used herein, close contact means, for example, having been within 6 feet of an individual infected with SARS-CoV-2 for a total of at least 15 minutes, or a total of at least 1 hour, or over 24 hours, or suspected of having been. Close contacts with a confirmed COVID-19 human include, but are not limited to, subjects who have come into contact with, or are suspected of having come into contact with, one or more SARS-CoV-2 positive individuals in a daycare center, airplane, restaurant, school, medical institution (such as a doctor's office or hospital). Close contacts include, but are not limited to, healthcare workers or first responders (such as law enforcement officers, emergency medical technicians, nurses, doctors, or firefighters) who have come into contact with a human suspected of being infected with SARS-CoV-2.

[0020] In some embodiments, subjects are at risk of developing the disease caused by SARS-CoV-2 due to age, immunodeficiency, or one or more pre-existing conditions. For example, subjects at risk may be elderly (i.e., 65 years of age or older) or immunodeficiency due to disease (genetic or acquired disease) or immunosuppressant therapy. Subjects at risk of developing beta-coronavirus-related disease may also include those with pre-existing conditions, such as chronic health conditions, including but not limited to: obesity; diabetes (type 1 or type 2); AIDS; chronic lung conditions such as asthma, COPD, pulmonary fibrosis, cystic fibrosis, pulmonary hypertension, emphysema, and bronchitis; neutrophilia; a history of myocardial infarction or stroke; coronary artery disease; cardiomyopathy; dementia or Alzheimer's disease; chronic kidney or liver disease; organ transplantation; and one or more autoimmune diseases (e.g., SLE or RA). In some embodiments, subjects have a history of tobacco smoking.

[0021] Individuals with a weakened or impaired immune system include, but are not limited to, cancer patients, particularly those undergoing chemotherapy, radiation therapy, or immunotherapy; individuals who have received organ or tissue transplants; individuals taking medications to suppress the immune response (e.g., corticosteroids, cyclosporine, tacrolimus); individuals with HIV or AIDS; individuals with diabetes; individuals recovering from illness, surgery, or trauma; and individuals with malnutrition.

[0022] Prophylactic administration of NSAIDs and ketotifen is also suitable for individuals who cannot tolerate (or benefit from) vaccines. For example, many elderly individuals are known not to respond effectively to vaccines. In embodiments, prophylactic administration of the pharmaceutical compositions according to this disclosure includes administration before the subject is infected with SARS-CoV-2 or before SARS-CoV-2 is detected in the subject.

[0023] In some embodiments, the therapies according to the present disclosure are provided at the time of vaccination (one or more days) to inhibit inflammatory symptoms associated with the beta-coronavirus spike protein contained in or encoded by the vaccine.

[0024] In various embodiments, therapy may be initiated before or after the onset of symptoms. In some embodiments, therapy is initiated shortly after the first appearance of symptoms (i.e., immediately after the first appearance of symptoms). In some embodiments, subjects according to this disclosure are asymptomatic but test positive for SARS-CoV-2 (or other beta-coronavirus). As used herein, testing for SARS-CoV-2 (or other beta-coronavirus) includes, but is not limited to, genetic testing (e.g., PCR, or nucleic acid amplification-based testing), antigen testing, or antibody (i.e., serological) testing. In some embodiments, therapy according to this disclosure is initiated as soon as an individual is first suspected of having an infection or disease caused by SARS-CoV-2 (or other beta-coronavirus), but before the onset of symptoms of the disease. In various embodiments, subjects have one or more risk factors for COVID-19, as already described.

[0025] In various embodiments, subjects are experiencing one or more symptoms selected from the following: throat or nasal congestion, cough, fever, loss of taste, loss of smell, muscle pain, joint pain (myalgia), fatigue, sore throat, headache, myocarditis, coagulation disorder, kidney injury or failure, liver injury or failure, neurological disorder, hypoxia, pneumonia with or without hypoxia, acute respiratory syndrome, or organ failure.

[0026] In some embodiments, the therapy according to this disclosure is initiated during the incubation period of betacoronavirus or SARS-CoV-2, when the virus has replicated and established a residence within the subject, primarily within the respiratory system of the subject. Symptoms at this stage are usually mild and often nonspecific, and may include fever, cough, sore throat, diarrhea, loss of taste or smell, fatigue, and headache.

[0027] In some embodiments, the therapy according to this disclosure is initiated during a pulmonary phase which may include hypoxia and may require oxygen supplementation.

[0028] In further embodiments, the therapy according to this disclosure is initiated in subjects experiencing extrapulmonary systemic hyperinflammatory syndrome. For example, a subject may exhibit fervent inflammation resulting from high levels of circulating cytokines such as interleukin (IL)-6 and tumor necrosis factor alpha (TNF-α). In some embodiments, a subject may experience acute respiratory syndrome and / or multiple organ failure.

[0029] In some embodiments, the therapies according to this disclosure are administered to subjects exhibiting early symptoms of COVID-19. Examples of early symptoms of COVID-19 include, but are not limited to, respiratory symptoms (without substantial hypoxia), throat and nasal congestion, cough, fever, loss of taste or smell (osmoticism), muscle pain, joint pain (myalgia), fatigue, sore throat, or headache.

[0030] In some embodiments, the therapies according to this disclosure are administered to subjects having symptoms of long COVID, or the therapies according to this disclosure prevent or reduce the likelihood of long COVID. As used herein, long COVID refers to a series of new or ongoing symptoms that may last for weeks or months after a subject has been initially infected with SARS-CoV-2 or has experienced symptoms of COVID-19. Long COVID may occur in subjects who have had COVID-19, even if the illness was mild or there were no initial symptoms. Symptoms of long COVID include difficulty breathing or shortness of breath, fatigue or exhaustion, symptoms that worsen after physical or mental activity, difficulty thinking or difficulty concentrating (sometimes referred to as “brain fog”), cough, chest or stomach pain, headache, rapid or pounding heart (i.e., heart palpitations), joint or muscle pain, diarrhea, sleep disturbances, fever, dizziness when standing (lightheadedness), rash, mood changes, changes in smell or taste, and changes in the menstrual cycle.

[0031] NSAIDs as disclosed herein include, but are not limited to, aspirin (i.e., acetylsalicylic acid), ibuprofen (i.e., isobutylphenylpropanoic acid), naproxen (i.e., 6-methoxy-α-methyl-2-naphthaleneacetic acid), diclofenac (2-[(2,6-dichlorophenyl)-amino]benzeneacetic acid), diflunisal (2',4'-difluoro-4-hydroxybiphenyl-3-carboxylic acid), etodrac ((1,8-diethyl-1,3,4,9-tetrahydropyrano[3,4-b]indole-1-yl)acetic acid, indomethacin (2-{l-[(4-chlorophenyl)-carbonyl]-5-methoxy-2-methyl-1H-indole-3-yl}acetic acid), ketoprofen (3-benzoyl-α-methylbenzeneacetic acid), and ketrolac (2-amino-2-(hydroxymethyl)-l,3-propane Diol), meloxicam (4-hydroxy-2-methyl-N-(5-methyl-2-thiazolyl)-2H-1,2-benzothiazine-3-carboxamide-1,1-dioxide), nabumetone (4-(6-methoxy-2-naphthyl)-2-butanone), oxaprozin (3-(4,5-diphenyl-l,3-oxazole-2-yl)propionic acid), piroxicam (4-hydroxy-2-methoxy-N-2-pyridinyl It contains (-2H-l,2-benzothiazine-3-carboxamide 1,1-dioxide), salsalate (2-(2-hydroxybenzoyl)-oxybenzoic acid), sulindac ({(lZ)-5-fluoro-2-methyl-l-[4-(methylsulfinyl)-benzylidene]-lH-inden-3-yl}acetic acid), and tolmetin ([l-methyl-5-(4-methylbenzoyl)-lH-pyrrole-2-yl]acetic acid).

[0032] In certain embodiments, the NSAID comprises naproxen (6-methoxy-α-methyl-2-naphthaleneacetic acid) and / or indomethacin (2-{l-[(4-chlorophenyl)-carbonyl]-5-methoxy-2-methyl-lH-indole-3-yl}acetic acid). In some embodiments, the present disclosure provides a pharmaceutical composition comprising indomethacin, naproxen, and ketotifen.

[0033] As used herein, NSAIDs (e.g., naproxen and indomethacin) or ketotifen include all pharmaceutically acceptable versions thereof, for example, all pharmaceutically acceptable salts, stereoisomers and / or any mixtures thereof, all pharmaceutically acceptable amphoteric and / or any mixtures thereof, all pharmaceutically acceptable polymorphic forms and / or any mixtures thereof, and all pharmaceutically acceptable complexes (including solvates) and / or any mixtures thereof. Salts include their racemates, enantiomers, or any mixtures thereof. Particularly preferred salts include alkali metal salts (e.g., sodium and / or potassium salts), alkaline earth metal salts (e.g., magnesium and / or calcium salts), aluminum salts, ammonium salts, salts of suitable organic bases (e.g., alkylamines and / or methyl-D-glutamine salts), and salts of amino acids (e.g., arginine and / or lysine salts). Salts also include all enantiomer salts formed with pharmaceutically acceptable chiral acids and / or bases and / or any mixture of enantiomers of such salts (e.g., any mixture of those including (+) tartrates, (-) tartrates, and / or racemic mixtures). Typical examples of salts include naproxen sodium, indomethacin sodium, and ketotifen fumarate.

[0034] As used herein, “treating” a disease caused by SARS-CoV-2 means reducing, eliminating, or preventing at least one symptom of the disease, or reducing or eliminating the presence or activity of SARS-CoV-2 infection in a subject. Treatment includes the prevention and / or reduction of symptoms and / or morbidity associated with the disease. Treatment is effective in preventing or reducing symptoms associated with a disease caused by beta-coronavirus or SARS-CoV-2, particularly in reducing or preventing symptoms such as throat and nasal congestion, cough, fever, loss of taste, loss of smell, muscle pain, arthralgia (myalgia), fatigue, sore throat, headache, hypoxia, pneumonia with or without hypoxia, severe acute respiratory syndrome, or organ failure. Furthermore, treatment is effective in shortening the duration or severity of symptoms of the disease.

[0035] As used herein, “preventing” disease caused by SARS-CoV-2 means preventing a subject from becoming infected with SARS-CoV-2, or reducing or eliminating the presence or activity of SARS-CoV-2 in the subject, thereby preventing the development or progression of disease in a subject infected with SARS-CoV-2.

[0036] The method of the present invention involves administering one or more pharmaceutical compositions to a human subject in an amount effective in treating or preventing beta-coronavirus infection or related disease in the subject. NSAIDs and ketotifen may be administered as a single unit dose (composition) or in separate compositions. Administration may be by a physician or by self-administration. The actual amount of the active agent in a particular case may vary depending on the specific formulation, mode of application, specific application site, and the subject being treated (e.g., age, sex, size, drug resistance, etc.).

[0037] In some embodiments, the methods of the present disclosure include administering an effective amount of indomethacin and ketotifen. In certain embodiments, indomethacin and ketotifen are administered at least once daily, with the daily dose of indomethacin being about 20 mg to about 300 mg, about 40 mg to about 200 mg, or about 75 mg to about 150 mg. In various embodiments, the daily dose of ketotifen is about 1 mg to about 10 mg, or about 1 mg to about 6 mg, or about 2 mg to about 5 mg, or about 3 mg to about 4 mg.

[0038] In some embodiments, the methods of the present disclosure include administering naproxen and ketotifen. In certain embodiments, naproxen and ketotifen are administered at least once daily, with daily doses of naproxen ranging from approximately 100 mg to approximately 1600 mg, approximately 200 mg to approximately 1400 mg, approximately 400 mg to approximately 1200 mg, approximately 600 mg to approximately 1000 mg, or approximately 700 mg to approximately 900 mg. In various embodiments, daily doses of ketotifen ranging from approximately 1 mg to approximately 10 mg, or approximately 1 mg to approximately 6 mg, or approximately 2 mg to approximately 5 mg, or approximately 3 mg to approximately 4 mg.

[0039] In certain embodiments, the method of the Disclosure involves administering a pharmaceutical composition comprising an effective amount of indomethacin in doses of about 20 mg to about 300 mg, about 40 mg to about 200 mg, or about 75 mg to about 150 mg, and ketotifen in doses of about 1 mg to about 6 mg, about 2 mg to about 5 mg, or about 3 mg to about 4 mg. In some embodiments, the Disclosure involves administering a pharmaceutical composition comprising ketotifen and at least one NSAID in a unit dose. In certain embodiments, the unit dose of the Disclosure comprises about 50 mg of indomethacin and about 1 mg of ketotifen, and the unit dose is administered two or three times a day.

[0040] In some embodiments, the disclosure provides pharmaceutical compositions comprising an NSAID (e.g., indomethacin and / or naproxen) and ketotifen as active pharmaceutical ingredients. In some embodiments, the pharmaceutical composition comprises an NSAID (e.g., indomethacin and / or naproxen) and ketotifen as active pharmaceutical ingredients. In these embodiments, the composition may include inactive ingredients, such as pharmaceutical excipients, sweeteners, flavorings, taste masking ingredients, diluents, alum, stabilizers, buffers, colorants, respiratory neutralizers (e.g., peppermint or menthol scents), emulsifiers, suspending agents, and agents for controlling delivery. The compositions in some embodiments may have additional ingredients (including active ingredients) that do not affect the basic and novel properties of the composition. Such components that may be specifically excluded in some embodiments include cyclooxygenase-2-selective inhibitors (i.e., COX-2-selective inhibitors) or their prodrugs, adrenergic agonists, anticholinergics, zinc (e.g., zinc acetate, zinc gluconate, zinc sulfate), preconalil, phosphordiesterase type 4 modulators, recombinant human uterine globin, IL-9 antagonists or IL-8 antagonists (including blocking monoclonal antibodies or peptides), glycerophosphate calcium, sedative antihistamines, nasal decongestants, ginsenosides, and Examples include minoquinolone (e.g., chloroquine, hydroxycycloquine), protease inhibitors (e.g., lopinavir, ritonavir, nafamostat, and camostat), H2 receptor antagonists (e.g., famotidine), umifenovir, thiazolids (e.g., nitazoxanide), ivermectin, corticosteroids, tocilizumab, sarilumab, bevacizumab, selective serotonin reuptake inhibitors (e.g., fluvoxamine), remdesivir, actemra, baricitinib, dexamethasone (glucocorticoid), aviptadil, and azithromycin.

[0041] In some embodiments, the disclosure includes administering an NSAID (e.g., indomethacin and / or naproxen) and ketotifen at least once daily. In further embodiments, the NSAID and ketotifen are administered about once to about three times daily. In certain embodiments, the NSAID and ketotifen are administered for at least one week. In even further embodiments, the NSAID and ketotifen are administered for at least two weeks. In even further embodiments, the NSAID and ketotifen are administered for at least about three weeks, or about one month.

[0042] In some embodiments, the Disclosure provides pharmaceutical compositions comprising an NSAID and ketotifen, the compositions further comprising a pharmaceutical carrier, a pharmaceutical excipient, a sweetener, a flavoring agent, a taste masking agent, a diluent, alum, a stabilizer, a buffer, a coloring agent, a respiratory neutralizer (e.g., a peppermint or menthol scent), an emulsifier, a suspension agent, a spray agent, and combinations thereof.

[0043] Examples of daily doses administered in the method of the present invention are shown below. The daily dose can be administered in one dose or multiple doses, typically two doses. The quantity can be defined by a range and by lower and upper boundary ranges. Exemplary lower and upper limits are shown below, each of which may be taken as a separate element.

[0044] Ketotifen may be administered in doses of approximately 0.5 mg to 10 mg, with examples of lower limits within this range being approximately 1 mg, 3 mg, and 4 mg. Other examples of upper limits within this range are approximately 5 mg, 6 mg, and 2.8 mg. Typical doses are approximately 1 mg to 4 mg per day, typically administered as 1 to 2 mg twice daily.

[0045] Naproxen dosages range from approximately 110 mg to 1650 mg, with other lower limits including approximately 220 mg, 320 mg, 550 mg, and 720 mg. Other upper limits include approximately 880 mg, 1000 mg, 1300 mg, and 1500 mg. Typical dosages range from approximately 220 mg to 1100 mg per day, administered twice daily, more typically 220 mg of naproxen sodium twice daily, or 550 mg of naproxen sodium twice daily.

[0046] Indomethacin is administered in doses ranging from approximately 25 mg to 300 mg, with other lower limits including approximately 50 mg, 75 mg, 100 mg, and 125 mg. Other upper limits include approximately 150 mg, 175 mg, 200 mg, and 250 mg. A typical dosage is approximately 50–150 mg / day, usually administered as 25 mg three times a day or 50 mg three times a day.

[0047] According to embodiments of the present disclosure, the pharmaceutical composition(s) may be administered orally, intravenously, intramuscularly, subcutaneously, transdermally, by inhalation, or intranasally. In further embodiments, the composition comprising ketotifen and an NSAID, such as indomethacin and / or naproxen, may be combined in the form of tablets, capsules, lozenges, or chewing gum.

[0048] In further embodiments, the compositions of the present disclosure are administered by nebulizer or by powder or solution aerosol (aerosol spray, mist, or powder) for local delivery to the lungs. Such local delivery is useful for subjects already experiencing lung injury.

[0049] In embodiments, the compositions according to the present disclosure may be administered orally by any method known in the art. For example, the compositions may be administered in the form of, for example, orally soluble tablets, tablets including chewable tablets, capsules, lozenges, pills (e.g., Pastil, sugar-coated), troches, elixirs, suspensions, syrups, wafers, chewing gum, strips, films (e.g., orally soluble thin films), soluble powders, effervescent compositions, and the like. In some embodiments, the compositions are tablets or capsules for oral delivery (e.g., intra-enteral or oral delivery).

[0050] For tablets intended for oral use, commonly used carriers include lactose and corn starch, with lubricants such as magnesium stearate commonly added. For oral administration in capsule form, useful carriers include lactose and corn starch. Further examples of carriers and excipients include milk, sugars, certain types of clay, gelatin, stearic acid or its salts, calcium stearate, talc, vegetable fats or oils, gums, and glycols.

[0051] When an aqueous suspension is used for oral administration, emulsifiers and / or suspending agents are commonly added. In addition, sweeteners and / or flavoring agents may be added to the oral composition.

[0052] In additional embodiments, the compositions according to the present disclosure may be administered intestinally or parenterally, for example, intravenously, intramuscularly, subcutaneously, intraperitoneally, sublingually, or rectally (e.g., by suppositories) as injectable solutions or suspensions. For intramuscular, intraperitoneal, subcutaneous, and intravenous use, sterile solutions of the pharmaceutical composition may be used, and the pH of the solution may be suitably adjusted and buffered. For intravenous use, the total concentration of solute(s) may be controlled to make the preparation isotonic.

[0053] In certain embodiments, NSAIDs and ketotifen are provided as orally dissolvable tablets, with or without taste-masking agents, diluents, etc. Such tablets can be administered on the tongue without water, dissolve quickly, and are absorbed into the gastrointestinal tract or oral cavity. Orally dissolvable tablets can be formulated by several techniques, including compression and lyophilization, as is known to those skilled in the art.

[0054] In some embodiments, the composition is a lozenge or troche containing an NSAID and ketotifen, with or without taste-masking components, diluents, etc. Such lozenges / troches can be administered without water and can be slowly dissolved in the mouth or swallowed or chewed. Such lozenges / troches can be formulated by compression, as is known to those skilled in the art.

[0055] In various embodiments, the pharmaceutical composition is a controlled-release composition. Controlled-release drug delivery achieves a certain level of drug over a specific period of time. The level is typically measured by plasma concentration. Methods for controlled-release of drugs are well known in the art. An example of a controlled-release formulation is a capsule containing beads, some of which dissolve instantaneously, and some of which dissolve with a delay time due to different types of bead coatings.

[0056] This disclosure includes the embodiments described in the following sections. [Section 1] A method for treating or preventing beta-coronavirus infection or related disease in a person requiring such treatment, comprising administering a non-steroidal anti-inflammatory drug (NSAID) and ketotifen. [Section 2] The method according to item 1, wherein the subject is a mammal. [Section 3] The method described in item 2, wherein the subject is a human. [Section 4] The method according to any one of items 1 to 3, wherein the beta-coronavirus is SARS-CoV-2. [Section 5] The method according to item 4, wherein the subject is negative for SARS-CoV-2 infection. [Section 6] The method according to item 5, wherein the subject has had close contact with one or more individuals who have tested positive for SARS-CoV-2. [Section 7] The method according to paragraph 6, wherein the subject is a healthcare professional or a first responder. [Section 8] The method described in item 4, wherein the subject has previously tested positive for SARS-CoV-2. [Section 9] The method according to item 8, wherein the SARS-CoV-2 is an alpha, beta, gamma, delta, or lambda variant. [Section 10] The method according to any one of paragraphs 1 to 9, wherein the subject is over 65 years of age and / or is immunocompromised due to immunosuppressant therapy and / or has a chronic health condition. [Section 11] The method according to paragraph 10, wherein the aforementioned chronic health condition is selected from one or more of the following: obesity; diabetes; AIDS; chronic lung conditions, optionally selected from asthma, COPD, pulmonary fibrosis, cystic fibrosis, pulmonary hypertension, emphysema, and bronchitis; neutrophilia; a history of myocardial infarction or stroke; coronary artery disease; congestive heart failure; cardiomyopathy; dementia or Alzheimer's disease; chronic kidney or liver disease; organ transplantation; and autoimmune diseases. [Section 12] The method according to any one of items 8 to 11, wherein the SARS-CoV-2 related disease in question is asymptomatic. [Section 13] The method described in any one of paragraphs 8 to 11, wherein the subject is experiencing one or more symptoms of COVID-19. [Section 14] The method according to paragraph 13, wherein the symptoms include one or more of the following: throat or nasal congestion, cough, fever, loss of taste, loss of smell, muscle pain, joint pain (myalgia), fatigue, sore throat, headache, myocarditis, coagulation disorder, kidney injury or failure, liver injury or failure, neurological disorder, hypoxia, pneumonia with or without hypoxia, acute respiratory syndrome, or multiple organ failure. [Section 15] The method according to item 14, wherein the subject has symptoms of hypoxia and / or acute respiratory distress syndrome. [Section 16] The method according to item 14, wherein the subject has chronic headache and / or fatigue. [Section 17] The method according to item 14, wherein the subject has long COVID. [Section 18] The method according to any one of claims 1 to 17, wherein the NSAID and the ketotifen are administered in separate compositions. [Section 19] The method according to any one of claims 1 to 17, wherein the NSAID and the ketotifen are administered in the same composition. [Section 20] The method according to claim 18 or 19, wherein the composition is administered orally, intravenously, as an inhalation solution or powder aerosol, intramuscularly, subcutaneously, percutaneously, or intranasally. [Section 21] The method according to claim 20, wherein the composition is administered by a nebulizer. [Section 22] The method according to claim 20, wherein the composition is in the form of a tablet, capsule, lozenge, or chewing gum. [Section 23] The method according to any one of claims 20 to 22, wherein the composition essentially consists of the NSAID and the ketotifen as active pharmacokinetic components. [Section 24] The method according to claim 23, wherein the composition comprises the NSAID and the ketotifen as active pharmacokinetic components. [Section 25] The method according to any one of claims 20 to 24, wherein the composition further comprises a pharmaceutical carrier, a pharmaceutical excipient, a sweetener, a flavoring agent, a taste masking component, a diluent, alum, a stabilizer, a buffer, a coloring agent, a respiratory neutralizer, an emulsifier, a suspension agent, a spray agent, and combinations thereof. [Section 26] The method according to any one of claims 1 to 25, wherein the NSAID and the ketotifen are administered before the onset of COVID symptoms. [Section 27] The method according to any one of claims 1 to 25, wherein the NSAID and the ketotifen are administered after the onset of COVID symptoms. [Section 28] The method according to item 27, wherein the NSAID and the ketotifen are optionally administered during the initial infection phase at the time of the first appearance of symptoms of COVID-19. [Section 29] The method according to any one of claims 1 to 25, wherein the NSAID and the ketotifen are administered at the time of vaccination against beta-coronavirus or SARS-CoV-2. [Section 30] The method according to any one of claims 1 to 29, wherein the NSAID and the ketotifen are administered at least once daily. [Section 31] The method according to item 30, wherein the NSAID and the ketotifen are administered about once to about three times a day. [Section 32] The method according to claim 30 or 31, wherein the NSAID and the ketotifen are administered for at least one week. [Section 33] The method according to claim 32, wherein the NSAID and the ketotifen are administered for at least two weeks. [Section 34] The method according to claim 33, wherein the NSAID and the ketotifen are administered for at least three weeks or about one month. [Section 35] The method according to any one of claims 1 to 34, wherein the NSAID is selected from indomethacin, naproxen, aspirin, ibuprofen, diclofenac, diflunisal, etodlach, ketoprofen, ketrolac, meloxicam, nabumetone, oxaprozin, piroxicam, salsalate, sulindac, and tolmetin, or a combination thereof. [Section 36] The method according to item 35, wherein the NSAID is indomethacin. [Section 37] The method according to item 36, wherein the daily dose of indomethacin is approximately 20 mg to approximately 300 mg, approximately 40 mg to approximately 200 mg, or approximately 75 mg to approximately 150 mg. [Section 38] The method according to item 35, wherein the NSAID is naproxen. [Section 39] The method according to item 38, wherein the daily dose of naproxen is approximately 100 mg to approximately 1600 mg, approximately 200 mg to approximately 1400 mg, approximately 400 mg to approximately 1200 mg, approximately 600 mg to approximately 1000 mg, or approximately 700 mg to approximately 900 mg. [Section 40] The method according to any one of claims 35 to 39, wherein the daily dose of ketotifen is approximately 1 mg to approximately 10 mg. [Section 41] The method according to item 40, wherein the daily dose of ketotifen is approximately 1 mg to approximately 6 mg. [Section 42] The method according to item 41, wherein the daily dose of ketotifen is approximately 2 mg to approximately 5 mg. [Section 43] The method according to item 42, wherein the daily dose of ketotifen is approximately 3 mg to approximately 4 mg. [Section 44] The method according to any one of claims 35 to 43, wherein the NSAID and the ketotifen are combined in a unit dose. [Section 45] The method according to claim 44, wherein the NSAID is indomethacin, the amount of indomethacin in the unit dose is about 50 mg, the amount of ketotifen is about 1 mg, and the unit dose is administered two or three times a day. [Section 46] The method according to any one of paragraphs 1 to 45, wherein the method reduces the viral load in the subject and / or prevents or reduces the symptoms of the disease. [Section 47] A pharmaceutical composition for use in preventing or treating beta-coronavirus infection or related diseases, wherein the pharmaceutical composition comprises a nonsteroidal anti-inflammatory drug (NSAID) and ketotifen. [Section 48] The pharmaceutical composition according to claim 47, wherein the composition is formulated for oral, intravenous, aerosol, intramuscular, subcutaneous, transdermal, or intranasal administration. [Section 49] The pharmaceutical composition according to claim 48, wherein the composition is formulated for administration by nebulizer. [Section 50] The pharmaceutical composition according to claim 49, wherein the composition is in the form of a tablet, capsule, lozenge, or chewing gum. [Section 51] The pharmaceutical composition according to any one of claims 47 to 50, wherein the composition essentially consists of the NSAID and the ketotifen as active pharmaceutical components. [Section 52] The pharmaceutical composition according to claim 51, wherein the composition comprises the NSAID and the ketotifen as active pharmaceutical components. [Section 53] The pharmaceutical composition according to any one of claims 48 to 52, wherein the composition further comprises a pharmaceutical carrier, a pharmaceutical excipient, a sweetener, a flavoring agent, a taste masking component, a diluent, alum, a stabilizer, a buffer, a coloring agent, a respiratory neutralizer, an emulsifier, a suspension agent, a spray agent, and combinations thereof. [Section 54] The pharmaceutical composition according to any one of claims 48 to 53, wherein the NSAID is selected from indomethacin, naproxen, aspirin, ibuprofen, diclofenac, diflunisal, etodolac, ketoprofen, ketrolac, meloxicam, nabumetone, oxaprozin, piroxicam, sarsalate, sulindac, and tolmetine, or a combination thereof. [Section 55] The pharmaceutical composition according to item 54, wherein the NSAID is indomethacin. [Section 56] The pharmaceutical composition according to item 55, wherein the daily dose of indomethacin is approximately 20 mg to approximately 300 mg, approximately 40 mg to approximately 200 mg, or approximately 75 mg to approximately 150 mg. [Section 57] The pharmaceutical composition according to item 54, wherein the NSAID is naproxen. [Section 58] The pharmaceutical composition according to item 57, wherein the daily dose of naproxen is approximately 100 mg to approximately 1600 mg, approximately 200 mg to approximately 1400 mg, approximately 400 mg to approximately 1200 mg, approximately 600 mg to approximately 1000 mg, or approximately 700 mg to approximately 900 mg. [Section 59] The pharmaceutical composition according to any one of claims 54 to 58, wherein the daily dose of ketotifen is approximately 1 mg to approximately 10 mg. [Section 60] The pharmaceutical composition according to item 59, wherein the daily dose of ketotifen is approximately 1 mg to approximately 6 mg. [Section 61] The pharmaceutical composition according to item 60, wherein the daily dose of ketotifen is approximately 2 mg to approximately 5 mg. [Section 62] The pharmaceutical composition according to item 61, wherein the daily dose of ketotifen is approximately 3 mg to approximately 4 mg. Other aspects and embodiments of this disclosure will become apparent from the following exemplary examples. [Examples]

[0057] The 2019 coronavirus infectious disease (COVID-19) is caused by infection with a novel severe acute respiratory syndrome coronavirus (SARS-CoV-2). Treatment options for COVID-19 are limited. The objective of these studies was to demonstrate the efficacy of ketotifen, naproxen, and indomethacin, either alone or in combination, in reducing SARS-CoV-2 replication. In addition, the cytotoxicity of the drugs was evaluated. The findings demonstrate that both ketotifen and indomethacin or naproxen in combination reduce viral yield. Ketotifen alone (EC 50 Compared to ketotifen alone (60% inhibition), increases in the percentage of SARS-CoV-2 inhibition to 79%, 83%, and 93% were observed when co-administered with 25, 50, and 100 μM indomethacin, respectively. Compared to ketotifen alone, increases in the percentage of SARS-CoV-2 inhibition to 68%, 68%, and 92% were observed when co-administered with 25, 50, and 100 μM naproxen, respectively. For both drug combinations, the observations suggest additive or synergistic effects compared to the administration of the drugs alone. No cytotoxic effects were observed with respect to the administered doses of ketotifen, naproxen, and indomethacin.

[0058] Materials and methods Ketotifen, indomethacin, and naproxen were obtained from Sigma Aldrich, and remdesivir from MedChemExpress. SARS-CoV-2 hCoV-19 / Australia / VIC01 / 2020 was provided by the Peter Doherty Institute for Infection and Immunity in Melbourne. African green monkey kidney (Vero E6) cells were obtained from ATCC (ATCC-CRL1586) (Noble Park North, VIC, Australia).

[0059] The viral stock was expanded via passage within Vero E6 cells in growth medium, which consisted of minimal essential medium without L-glutamine, supplemented with 1% (w / v) L-glutamine, 1.0 μg / mL TPCK-trypsin, 0.2% BSA, 1 × Pen / Strep, and 1% insulin transferrin selenium (ITS).

[0060] African green monkey kidney (Vero E6) cells (ATCC-CRL1586) were subcultured to generate a cell bank stock in cell growth medium. The cell growth medium consisted of l-glutamine-free minimal essential medium (MEM) supplemented with 10% (v / v) heat-inactivated fetal bovine serum (FBS) and 1% (w / v) l-glutamine. After subculturing the Vero E6 cells for up to 13 passages, a fresh working cell bank stock was recovered from liquid nitrogen for further use.

[0061] Cytopathic effect (CPE) assay The cytopathic effects of indomethacin, ketotifen, and naproxen were compared with those of remdesivir (positive control).

[0062] Vero E6 cells were seeded in 100 μL of seeding medium (MEM supplemented with 1% (w / v) l-glutamine and 2% FBS) at a rate of 2 × 10⁶ 4 Cells were seeded in 96-well plates at the specified cell / well rate. The plates were incubated overnight at 37°C in 5% CO2. Test compounds were prepared fresh on the day of testing, vortexed, and visually inspected to confirm complete solubility.

[0063] The positive control compound remdesivir was prepared as a 10 mM stock in dimethyl sulfoxide (DMSO) and stored at -20°C. Compound dilutions were prepared on the day of the experiment. Eight DMSO dilution series, 3-fold for each of the four test compounds, were initially performed in the range of 100 mM to 0.045 mM. Intermediate dilution series in viral growth medium (MEM supplemented with 1% (w / v) l-glutamine, 2% FBS, and 8 μg / mL tosylphenylalanyl chloromethyl ketone (TPCK)-trypsin) were generated in the range of 800 μM to 0.37 μM. 50 μL volume from each compound's intermediate dilution series was added to triple wells of an assay plate pre-seed with Vero E6 cells. The DMSO concentration in the assay plate was maintained at 0.2%. One assay plate was prepared per compound.

[0064] Similarly, remdesivir was subjected to an initial DMSO dilution series and then to an intermediate dilution series to reduce the DMSO concentration in the assay plate to 0.2%. Remdesivir was tested at an initial concentration of 20 μM.

[0065] 50 μL of SARS-CoV-2 diluted in viral growth medium was added to the assay plate to generate a multiplicity of infection (moi) of 0.05. This moi had previously been determined to provide 100% cytopathic effect (CPE) after 4 days. To evaluate antiviral activity, the virus was added to three columns, and to evaluate cytotoxicity, virus-free viral growth medium was added to three columns.

[0066] The cytoprotective rates achieved by the positive control (remdesivir) and the test substance in virus-infected cells were calculated using the formula by Pauwels et al., as shown below (J. Virol. Methods 1988, 20, 309-321). Cell protection rate = ([ODt]virus - [ODc]virus / [ODc]mock - [ODc]virus) × 100 During the ceremony: [ODt] Virus = The optical density measured in a well that examines the effect of a given test substance or positive control at a concentration on virus-infected cells. [ODc] Virus = The optical density measured in a well that examines the effect of a negative control on virus-infected cells. [ODc] Mock = The optical density measured in a well that examines the effect of a negative control on mock-infected cells.

[0067] The EC 50 value was calculated from the cell protection rate results by non-linear regression analysis using the Hill (sigmoid Emax) equation. y = minimum y + (maximum y - minimum y ) / 1 + (EC 50 / X) D Where: X = the concentration of the test substance or control sample, Y = the cell protection rate, Min = minimum, Max = maximum, D = the slope coefficient.

[0068] Yield reduction assay Vero E6 cells were seeded in a 96-well plate at 2 × 10 4 cells / well in 100 μL of seeding medium (MEM supplemented with 1% (w / v) L-glutamine and 2% FBS). The plate was incubated overnight at 37 °C and 5% CO2.

[0069] For combination studies, the compounds were added together in a deep well plate at a 1:1 ratio and then a 100 μL volume of the compound combination was added to the cell monolayer. A 200 μL volume of virus (B3) was added to the plate (moi 0.003).

[0070] After 48 hours of incubation, the virus was harvested at each concentration and diluted 1:10 (n=1) or 1:100 (n=2) in virus growth medium. 100 microliters were added to three wells of a 96-well plate containing Vero E6 cells, and the entire plate was serially diluted threefold to obtain a total of nine different virus concentrations. Six of the wells contained assay medium alone (i.e., no virus) and were used as a control. The plates were incubated at 37°C for 3 days in a humidified 5% CO2 atmosphere, during which time CPE was induced. The cell monolayer was then microscopically observed using visual scoring of virus-induced CPE, which was used as the endpoint. TCID of the virus suspension 50 This was determined using the Reed-Muench method (Reed, LJ; Muench, HA simple method of estimating fifty percent endpoints. Am.J.Hyg. 1938, 27, 493-497). Virus yield was expressed as a percentage of viral growth compared to the case without drug addition, for each concentration.

[0071] Cytotoxicity assay The cytotoxic effects of indomethacin, ketotifen, and naproxen on Vero E6 cells were tested over 4 days or 48 hours to mimic either an antiviral assay or a yield reduction assay. The drugs were tested at the same concentrations used in Sections 2.1 and 2.2. Live cells were determined by staining with MTT or crystalline violet. 100 μL of 3 mg / mL MTT solution was added to a plate and incubated at 37°C for 2 hours in a 5% CO2 incubator. The wells were aspirated until dry, and formazan crystals were solubilized by adding 200 μL of 100% 2-propanol at room temperature for 30 minutes. Absorbance was measured on a plate reader at 540–650 nm. The medium was removed from the cells stained with crystalline violet and washed once with PBS. 40 μL of 0.25% crystalline violet / 20% methanol stain was added to each well and incubated at room temperature for 30 minutes. Crystalline violet stain was aspirated, the monolayer was washed 3-5 times with PBS, and the stain was solubilized by adding 1% sodium dodecyl sulfate (SDS) in 100 μL. After 30 minutes at room temperature, the absorbance was measured on a plate reader at 540-650 nm. 50% cytotoxic concentration (CC) 50 CC was defined as the concentration of the test compound that reduces the absorbance of mock-infected cells by 50% compared to the control value. 50 The value was calculated as follows: CC 50 =[ODt] mock / [ODc] mock.

[0072] result The results of the CPE assay are shown in Table 1. None of the treatments showed antiviral activity in the 4-day CPE assay.

[0073] [Table 1]

[0074] The results for ketotifen and naproxen in the Yield Reduction Assay (YRA) are summarized in Table 2 and Figure 2. Ketotifen alone inhibited SARS-CoV-2 and exhibited an EC50 of 48.9 μM. Naproxen alone did not inhibit SARS-CoV-2 and exhibited an EC50 greater than 100 μM. 50 It presented the following: EC 50 This is the effective concentration, i.e., the concentration at which viral infection is inhibited by 50%.

[0075] [Table 2]

[0076] The addition of ketotifen at a single concentration enhanced naproxen's ability to reduce viral yield. 50 The concentration decreased to less than 6 μM, which corresponds to approximately a 17-fold increase in the presence of ketotifen. The dose-response curve is shown in Figure 2. These data indicate additive or synergistic effects.

[0077] The results for ketotifen and indomethacin are summarized in Tables 3 and 4. Table 3 shows the results for ketotifen alone (EC 50 Compared to 60% inhibition with indomethacin alone, increases in the inhibition percentage of SARS-CoV-2 to 79%, 83%, and 93% were observed when co-administered with 25, 50, and 100 μM indomethacin, respectively, demonstrating a synergistic effect.

[0078] [Table 3]

[0079] [Table 4]

[0080] Indomethacin alone inhibited SARS-CoV-2, and 100.1 μM EC 50 This was shown (see Table 4). Indomethacin was EC2 in the presence of 50 μM ketotifen.50 Although it did not reach 60%, 60% inhibition was observed at 6.25 μM, and if the dilution series covered a lower range, EC 50 It was indicated that this would likely be reached at approximately 3 μM. These in vitro data suggest that lower doses of indomethacin are required to inhibit viral replication in the presence of 50 μM ketotifen, and that these doses exhibit additive or synergistic effects. Indomethacin inhibits EC in the presence of 25 μM, 12.5, 6.25, or 3.1 μM ketotifen. 50 It did not reach that goal.

[0081] The addition of ketotifen at a single concentration enhanced the ability of indomethacin to reduce viral yield. 50 The concentration decreased to less than 6 μM, which corresponds to approximately 13 times the effect in the presence of ketotifen. Figure 3 shows the dose-response curves for indomethacin and ketotifen alone and in combination. The dose-response curves suggest an additive or synergistic antiviral effect against SARS-CoV-2 when ketotifen and indomethacin are administered in combination.

[0082] The results of the cytotoxicity assessment are summarized in Table 5. 50% cytotoxic concentration (CC) 50 ) was defined as the compound concentration required to reduce cell viability by 50%. No cytotoxicity was observed for indomethacin, naproxen, and ketotifen at the concentrations tested.

[0083] [Table 5]

[0084] Current findings indicate that both ketotifen / indomethacin and ketotifen / naproxen combinations reduce viral yield. Ketotifen alone (EC 50Compared to 60% inhibition with indomethacin alone, increases in the inhibition percentage of SARS-CoV-2 to 90%, 97%, and 94% were observed when co-administered with 25, 50, and 100 μM indomethacin, respectively, indicating additive or synergistic effects. 50 It decreased to less than 6 μM, which corresponds to approximately 13 times the effect in the presence of ketotifen. Ketotifen alone (EC 50 Compared to 60% inhibition with ketotifen alone, increases in the percentage of SARS-CoV-2 inhibition to 68%, 68%, and 92% were observed when co-administered with naproxen at 25, 50, and 100 μM, respectively. This also indicates an additive or synergistic effect. Finally, no cytotoxic effects were observed for the concentrations of ketotifen, indomethacin, and naproxen tested.

[0085] conclusion These examples demonstrate that combinations of ketotifen with indomethacin or naproxen significantly reduce viral yield. Compared to ketotifen alone, the combination with indomethacin or naproxen exhibits additive or synergistic effects. No cytotoxic effects were observed at the concentrations of ketotifen, naproxen, and indomethacin tested.

[0086] Further considerations Where used herein, the phrase “at least one of” preceding a set of items, accompanied by the terms “and” or “or” to distinguish the set of items, modifies the entire list rather than each member of the list (i.e., each item). The phrase “at least one of” does not require a selection of at least one of each listed item; rather, it allows the meaning to include at least one of any one of the items, and / or at least one of any combination of the items, and / or at least one of each of the items. For example, the phrases “at least one of A, B, and C” or “at least one of A, B, or C” refer, respectively, to A only, B only, or C only, any combination of A, B, and C, and / or at least one of each of A, B, and C.

[0087] Where used herein, the term "approximately" means ±10% of the relevant number unless the context requires otherwise.

[0088] As used herein, the term “comprising” indicates the presence of a specified feature but allows for the possibility of other unspecified features. The term does not imply any particular proportion of the specified feature. The terms “comprising,” for example, “comprise,” and “comprises,” have correspondingly similar meanings.

[0089] The term “exemplary” is used herein to mean “serving as an example, illustration, or representation.” No embodiment described herein as “exemplary” should necessarily be construed as being preferable or advantageous to any other embodiment.

[0090] References to singular elements, unless otherwise specified, are not intended to mean "one and only one," but rather "one or more." All structural and functional equivalents to elements of the various configurations described herein, whether known to those skilled in the art or to be known thereafter, are expressly incorporated herein by reference and are intended to be encompassed by the subject art.

Claims

1. A combination pharmaceutical for treating or preventing beta-coronavirus infection in a target population, comprising a non-steroidal anti-inflammatory drug (NSAID) and ketotifen, wherein the NSAID is selected from naproxen or indomethacin.

2. The combination pharmaceutical according to claim 1, wherein the beta coronavirus is SARS-CoV-2.

3. The combination pharmaceutical according to claim 2, wherein the subject is negative for SARS-CoV-2 infection, and the subject has had close contact with one or more SARS-CoV-2 positive individuals.

4. The combination pharmaceutical according to claim 2, wherein the subject has previously tested positive for SARS-CoV-2.

5. The combination pharmaceutical according to any one of claims 1 to 4, wherein the subject is over 65 years of age and / or is immunocompromised due to immunosuppressant therapy of any choice and / or has a chronic health condition.

6. The combination pharmaceutical according to claim 5, wherein the chronic health condition is selected from one or more of the following: obesity; diabetes; AIDS; chronic lung conditions selected at will from asthma, COPD, pulmonary fibrosis, cystic fibrosis, pulmonary hypertension, emphysema, and bronchitis; neutrophilia; a history of myocardial infarction or stroke; coronary artery disease; congestive heart failure; cardiomyopathy; dementia or Alzheimer's disease; chronic kidney or liver disease; organ transplantation; and autoimmune diseases.

7. The combination pharmaceutical according to any one of claims 4 to 6, wherein the SARS-CoV-2 in question is asymptomatic.

8. The combination pharmaceutical according to any one of claims 4 to 6, wherein the subject has experienced one or more symptoms of COVID-19.

9. The combination pharmaceutical product according to claim 8, wherein the symptoms include one or more of the following: throat or nasal congestion, cough, fever, loss of taste, loss of smell, muscle pain, joint pain (myalgia), fatigue, sore throat, headache, myocarditis, coagulation disorder, kidney injury or failure, liver injury or failure, neurological disorder, hypoxia, pneumonia with or without hypoxia, acute respiratory syndrome, or multiple organ failure.

10. The combination pharmaceutical according to claim 9, wherein the subject has long COVID.

11. The combination pharmaceutical according to any one of claims 1 to 10, wherein the NSAID and the ketotifen are administered in separate compositions.

12. The combination pharmaceutical according to any one of claims 1 to 10, wherein the NSAID and the ketotifen are administered in the same composition.

13. The combination pharmaceutical according to claim 11 or 12, wherein the composition is administered orally, intravenously, as an inhaled solution or powder aerosol, intramuscularly, subcutaneously, transdermally, or intranasally.

14. The combination pharmaceutical according to claim 13, wherein the composition is in the form of a tablet, capsule, lozenge, or chewing gum.

15. The combination pharmaceutical according to claim 13 or 14, wherein the composition comprises the NSAID and the ketotifen as active pharmacochemical components.

16. The combination pharmaceutical product according to any one of claims 1 to 15, wherein the NSAID and the ketotifen are administered after the onset of COVID symptoms.

17. The combination pharmaceutical according to claim 16, wherein the NSAID and the ketotifen are optionally administered during the initial infection phase at the time of the first appearance of COVID-19 symptoms.

18. The combination pharmaceutical according to any one of claims 1 to 15, wherein the NSAID and the ketotifen are administered at the time of vaccination against beta-coronavirus or SARS-CoV-2.

19. The combination pharmaceutical according to any one of claims 1 to 18, wherein the NSAID and the ketotifen are administered at least once a day.

20. The combination pharmaceutical according to claim 1, wherein the NSAID is indomethacin.

21. The combination pharmaceutical according to claim 20, wherein the daily dose of indomethacin is approximately 20 mg to approximately 300 mg, or approximately 40 mg to approximately 200 mg.

22. The combination drug according to claim 20, wherein the daily dose of indomethacin is about 40 mg to about 200 mg.

23. The combination pharmaceutical according to claim 1, wherein the NSAID is naproxen.

24. The combination pharmaceutical according to claim 23, wherein the daily dose of naproxen is approximately 100 mg to approximately 1600 mg, approximately 200 mg to approximately 1400 mg, approximately 400 mg to approximately 1200 mg, or approximately 600 mg to approximately 1000 mg.

25. The combination pharmaceutical according to claim 23, wherein the daily dose of naproxen is approximately 600 mg to approximately 1000 mg.

26. The combination pharmaceutical according to any one of claims 20 to 25, wherein the daily dose of ketotifen is about 1 mg to about 10 mg.

27. The combination pharmaceutical according to any one of claims 20 to 26, wherein the NSAID and the ketotifen are combined in a unit dose.

28. The combination pharmaceutical according to claim 27, wherein the NSAID is indomethacin, the amount of indomethacin in the unit dose is about 50 mg, the amount of ketotifen is about 1 mg, and the unit dose is administered two or three times a day.

29. A pharmaceutical composition for use in preventing or treating betacoronavirus infection, wherein the pharmaceutical composition comprises a nonsteroidal anti-inflammatory drug (NSAID) and ketotifen, wherein the NSAID is indomethacin or naproxen.

30. The pharmaceutical composition according to claim 29, wherein the NSAID is indomethacin.

31. The daily dose of ketotifen is approximately 1 mg to approximately 10 mg, When the NSAID is indomethacin, the daily dose of indomethacin is approximately 40 mg to approximately 200 mg. The pharmaceutical composition according to claim 29 or 30, wherein, when the NSAID is naproxen, the daily dose of naproxen is about 600 mg to about 1000 mg.

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