Methods for treating coronavirus infection

Abstract

Compositions and methods for treating a subject for coronavirus infection are provided. The methods typically involve administering an effective amount of probenecid, its metabolite or analog, or a pharmaceutically acceptable salt thereof to the subject. The methods can be therapeutic and / or prophylactic. The amount of probenecid, its metabolite or analog, or a pharmaceutically acceptable salt thereof can be effective, for example, to reduce viral replication, alleviate one or more symptoms of a virus-related disease, disorder, or illness, or a combination thereof. In a preferred embodiment, the virus is a severe acute respiratory syndrome-related coronavirus such as SARS-CoV-2 or SARS-CoV, a Middle East respiratory syndrome-related coronavirus such as MERS-CoV, or a coronavirus that causes the common cold.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 008,624, filed Apr. 10, 2020; U.S. Provisional Patent Application No. 63 / 023,021, filed May 11, 2020; U.S. Patent Application No. 16 / 875,487, filed May 15, 2020; European Patent Application No. 20202059.0, filed Oct. 15, 2020; and U.S. Provisional Patent Application No. 63 / 151,551, filed Feb. 19, 2021, each of which is hereby specifically incorporated by reference in its entirety.

[0002] Reference to a Sequence Listing The sequence listing, submitted as a text file named "UGA_2020_148_06_PCT.txt", created on Apr. 2, 2021 and having a size of 167,524 bytes, is hereby incorporated by reference in accordance with 37 C.F.R.§1.52(e)(5).

[0003] Field of the Invention The field of the present invention generally relates to compositions and methods for treating and preventing coronavirus - related diseases.

Background Art

[0004] Background of the Invention A seventh human coronavirus was recently identified in Wuhan, China (Coronaviridae Study Group of the International Committee on Taxonomy of Viruses, Nat Microbiol 2020. DOI:10.1038 / s41564-020-0695-z, WHO "Pneumonia of unknown cause - China", World Health Organization: Online, 2020). This virus, discovered in December 2019, was initially described as COVID-19, but is now classified as a beta-coronavirus within the same species as the severe acute respiratory syndrome coronavirus (SARS-CoV), which caused the 2002-2003 pandemic (Coronaviridae Study Group of the International Committee on Taxonomy of Viruses, Nat Microbiol 2020.DOI:10.1038 / s41564-020-0695-z; Ratia et al., Proc Natl Acad Sci USA 2008,105(42),16119-24.DOI:10.1073 / pnas.0805240105; Karim et al., MedRxiv,5(4):536-44(2020),DOI:10.1101 / 2020.05.26.20104497). Therefore, COVID-19 is now classified as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Although SARS-CoV-2 does not appear to be as deadly as SARS-CoV, it is spreading rapidly worldwide, according to the World Health Organization's situation report. The rapid spread of SARS-CoV-2, and its ability to cause death, particularly in the elderly or individuals with underlying conditions, has created an urgent need for antiviral therapies and vaccines to be used against the virus (CDC website, "CDC People at Risk for Serious Illness from COVID-19").

[0005] Antiviral chemotherapy drugs targeting the SARS-CoV-2 virus are not yet available, and drug-resistant strains are likely to emerge rapidly. As a result, there is a rapidly increasing need to identify new anti-SARS-CoV-2 therapies, particularly those that target host gene products necessary for viral replication, in order to reduce the potential for drug resistance.

[0006] The focus has been largely on vaccination to eliminate or control SARS-CoV-2 infection. However, vaccine effectiveness is affected by limited manufacturing capacity, cold chain storage requirements, breakthroughs in viral infection, limited efficacy data, viral shedding from asymptomatic immunized subjects, delays and / or phased vaccine rollout, vaccine fears, ongoing mutation / evolution of the virus, and the possibility of COVID-19 continuing to spread globally. Therefore, there remains a strong need for therapeutic options for the treatment and prevention of SARS-CoV-2 infection and COVID-19 disease.

[0007] Therefore, an object of the present invention is to provide compositions and methods for treating infections caused by coronaviruses, including but not limited to SARS-CoV-2, which cause severe acute respiratory syndrome. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Coronaviridae Study Group of the International Committee on Taxonomy of Viruses,Nat Microbiol 2020.DOI:10.1038 / s41564-020-0695-z [Non-Patent Document 2] WHO “Pneumonia of unknown cause-China” World Health Organization: Online, 2020 [Non-Patent Document 3] Coronaviridae Study Group of the International Committee on Taxonomy of Viruses,Nat Microbiol 2020.DOI:10.1038 / s41564-020-0695-z [Non-Patent Document 4] Ratia et al., Proc Natl Acad Sci USA 2008,105(42),16119-24.DOI:10.1073 / pnas.0805240105 [Non-Patent Document 5] Karim et al., MedRxiv,5(4):536-44(2020),DOI:10.1101 / 2020.05.26.20104497 [Non-Patent Document 6] CDC website "CDC People at Risk for Serious Illness from COVID-19" [Overview of the Initiative] [Means for solving the problem]

[0009] Summary of the Invention Compositions and methods for treating subjects with coronavirus infection are provided. The method typically involves administering an effective amount of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof, to the subject. The amount of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof may be effective, for example, in reducing viral replication, or in alleviating one or more symptoms, or combination thereof, of a virus-related illness, disorder, or disease. Symptoms include, but are not limited to, fever, sinus and / or pulmonary congestion, runny nose or nasal congestion, cough, sneezing, sore throat, body aches, fatigue, shortness of breath, chest tightness, wheezing on exhalation, chills, muscle pain, headache, diarrhea, malaise, nausea, vomiting, and combinations thereof. The subject may be, for example, a mammal or a bird. In a preferred embodiment, the subject is a human.

[0010] The subjects may be symptomatic or asymptomatic. In some embodiments, the subjects are exposed to or will be exposed to the virus. In some embodiments, the treatment begins one, two, three, four, five hours or more, several days or weeks before or after exposure to the virus. In some embodiments, the subjects are not exposed to the virus. In some embodiments, the subjects anticipate being exposed to the virus. Thus, preventative and prophylactic methods are also provided and are included in the term “treatment.”

[0011] The virus may be Severe acute respiratory syndrome-related coronavirus, Bat Hp-betacoronavirus Zhejiang2013, Rousettus bat betavirus GCCDC1, Rousettus bat betavirus HKU9, Eidolon bat coronavirus C704, Pipistrellus bat coronavirus HKU5, Tylonycteris bar coronavirus HKU4, Middle East respiratory syndrome-related coronavirus, Hedgehog coronavirus, murine coronavirus, human coronavirus HKU 1, China Rattus coronavirus HKU24, betacoronavirus 1, Myodes coronavirus 2JL14, human coronavirus NL63, human coronavirus 229E, or human coronavirus OC43.

[0012] In a preferred embodiment, the virus is a severe acute respiratory syndrome-associated coronavirus, such as SARS-CoV-2, SARS-CoV, SARSr-CoV RaTG13, SARS-CoV PC4-227, or SARSr-CoV BtKY72.

[0013] In some embodiments, the virus is severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) having a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1 or 2.

[0014] In some embodiments, SARS-CoV-2 includes a spike protein having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 5. For example, SAR-CoV-2 may have one or more mutations compared to SEQ ID NO: 5. In some embodiments, the mutations are selected from H69, and possibly its deletion; V70, and possibly its deletion; G142, and possibly G142D; K417, and possibly K417N or K417T; E484, and possibly E484K; F486, and possibly F486L; and the N501 mutation, and possibly N501Y or N501T, as well as combinations thereof. SARS-CoV-2 may originate from, for example, lineages B.1.1.7, B.1.351, P.1, B.1.1.207, B.1.429, B.1.427, or B.1.525.

[0015] In some embodiments, the severe acute respiratory syndrome-associated coronavirus is a severe acute respiratory syndrome coronavirus (SARS-CoV), e.g., SAR-CoV, having a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 3.

[0016] In some embodiments, the virus is Middle East Respiratory Syndrome-related Coronavirus (MERS-CoV), for example, having a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with SEQ ID NO: 4.

[0017] In some embodiments, the subject has a disease or disorder associated with the virus. For example, in some embodiments, the subject has been exposed to or infected with SARS-CoV-2 and optionally has Coronavirus Disease 2019 (COVID-19). In some embodiments, the subject is not infected with influenza.

[0018] In some embodiments, the viral infection is detected or diagnosed in the subject before, during or after treatment. Detection and diagnosis of viral infection can include, but are not limited to, PCR tests designed to detect viral RNA, as well as serological and immunodiagnostic tests designed to detect antibodies against the virus. An exemplary test for SARS-CoV-2 infection / COVID-19 is the iAMP COVID-19 Detection Kit, which is a real-time fluorescent isothermal assay for use with raw samples without RNA extraction.

[0019] In some embodiments, the subject has been in close contact with a person who has a positive test for the virus or has COVID-19. Such a person may or may not exhibit one or more symptoms of the infection. In some embodiments, the subject of treatment is identified by contact tracing as having been exposed to the virus or as having been in contact with one or more persons infected with the virus.

[0020] Probencid, its metabolites or analogs, or pharmaceutically acceptable salts thereof are typically administered in a pharmaceutical composition comprising a pharmaceutically acceptable carrier and / or excipient. Accordingly, a pharmaceutical composition is also provided. Dosage forms are also provided, including but not limited to 500 mg tablets of probencid, its metabolites or analogs, or pharmaceutically acceptable salts thereof. In some embodiments, a subject is administered a dosage of 10 mg to 1,000 mg or 50 mg to 500 mg of probencid, its metabolites or analogs, or pharmaceutically acceptable salts thereof once, twice, three times, four times or five times a day. In some embodiments, the dosing regimen is a pulse dosing regimen that includes bolus administration of 1, 2, 3 or more large amounts (1,000 mg or more) in close proximity (e.g., at most 5, 10, 15, 30, 45 or 60 minutes, or 1, 2, 3, 4, 5, 6 hours or more) apart. In some embodiments, after the bolus administration, a drug-free holiday (e.g., at least 12 hours, or 1, 2, 3, 4, 5 or more days) continues until the drug level in the subject's serum has decreased to 0 or nearly 0 (e.g., 1%, 5%, 10% or 20% or less of the peak serum level), as needed.

[0021] Probencid, its metabolites or analogs, or pharmaceutically acceptable salts thereof can be administered systemically or topically. Exemplary routes of administration include, but are not limited to, oral, parenteral, topical or mucosal. In some embodiments, the composition is administered to the lungs (e.g., pulmonary administration) by oral inhalation or intranasal administration. In some embodiments, the composition is administered intranasally to the nasal mucosa.

Brief Description of the Drawings

[0022] [Figure 1] Figure 1 is a bar graph showing the effect of probencid (5 μM, 2.5 μM, 1 μM or 0.1 μM) compared to controls (DMSO (infected), DMSO (only)) on virus replication using a plaque reduction assay (from left to right).

[0023] [Figure 2] Figure 2 is a bar graph (from left to right) showing the effect of probenecid pretreatment (5 μM, 2.5 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, or 0.0001 μM) on viral replication compared to controls (DMSO (infection), DMSO (only)) using a plaque reduction assay.

[0024] [Figure 3] Figure 3 is a bar graph (from left to right) showing the effect of probenecid treatment (5 μM, 2.5 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, or 0.0001 μM) on viral replication compared to controls (DMSO (infection), DMSO (only)) using a plaque reduction assay.

[0025] [Figure 4] Figure 4 is a flowchart illustrating an assay for testing the prophylactic (pre-infection) and therapeutic (post-infection) effects of probenecid against SARS-CoV-2 in hamsters in vivo.

[0026] [Figure 5] Figure 5 is a line graph showing the body weight of hamsters in the control and experimental groups before and after treatment with probenecid and / or infection.

[0027] [Figure 6] Figure 6 is a bar graph showing TCID50 (Log TCID50 / ml) from the lungs of hamsters treated with probenecid.

[0028] [Figure 7] Figure 7 is a bar graph showing the results of a plaque assay (Log PFU / ml) from hamster lungs treated with probenecid.

[0029] [Figure 8A]Figure 8A is a bar graph showing the body mass index (BMI) of human subjects enrolled in an open-label, 28-day, investigator-initiated clinical trial (IIS) ("clinical trial") testing the efficacy of probenecid in subjects with mild to moderate SARS-CoV-2 infection. [Figure 8B] Figure 8B is a bar graph showing the age (years) of subjects enrolled in the clinical trial.

[0030] [Figure 9] Figure 9 is a bar graph showing the number of days until the negative SARS-CoV-2 test result for the subjects of the clinical trial.

[0031] [Figure 10] Figure 10 is a bar graph showing the improvement in symptoms in subjects during the clinical trial.

[0032] [Figure 11] Figure 11 is a graph showing the changes in body temperature in subjects during a clinical trial.

[0033] [Figure 12] Figure 12 is a graph showing the changes in high-sensitivity C-reactive protein (hs-CRP) in subjects during clinical trials.

[0034] [Figure 13] Figure 13 is a graph showing the changes in D-dimer levels in subjects during clinical trials.

[0035] [Figure 14] Figure 14 is a graph showing the changes in fibrinogen levels in subjects during clinical trials.

[0036] [Figure 15] Figure 15 is a graph showing the changes in lactate dehydrogenase (LDH) in subjects during clinical trials.

[0037] [Figure 16A-16C]Figures 16A-16C show (from left to right) 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM or 0.00001 μM of remdesivir, a virus control or cell control (16A), and 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM or 0.00001 μM of probenecid. This bar graph shows the mean responder intensity % of normal human bronchial epithelial (NHBE) cells infected with SARS-CoV-2, prophylactically treated with a viral control or cellular control (16B); or 0.1 μM probenecid + 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.00001 μM or 0.00001 μM remdesivir, viral control or cellular control (16C).

[0038] [Figures 17A-17C] Figures 17A-17C show (from left to right) 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM or 0.00001 μM of remdesivir, a virus control or cell control (17A), and 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM or 0.00001 μM of probenecid. This bar graph shows the mean responder intensity % of normal human bronchial epithelial (NHBE) cells infected with SARS-CoV-2, treated at the time of infection with a viral control or cell control (17B); or 0.1 μM probenecid + 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.00001 μM or 0.00001 μM remdesivir, viral control or cell control (17C).

[0039] [Figure 18A-18C]Figures 18A-18C show (from left to right) 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM of remdesivir, virus control or cell control (18A), 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM. This bar graph shows the mean responder intensity % of SARS-CoV-2 infected Vero cells prophylactically treated with M probenecid, viral control or cell control (18B); or 0.1 μM probenecid + 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM remdesivir, viral control or cell control (18C).

[0040] [Figures 19A-19C] Figures 19A-19C show (from left to right) 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM of remdesivir, virus control or cell control (19A), 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM. This bar graph shows the mean response intensity % of SARS-CoV-2 infected Vero cells treated at the time of infection with M probenecid, viral control or cell control (19B); or 0.1 μM probenecid + 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM remdesivir, viral control or cell control (19C).

[0041] [Figure 20A]Figure 20A is a bar graph showing the PFU / ml of SARS-CoV-2-infected normal human bronchial epithelial (NHBE) cells treated at the time of infection with remdesivir at concentrations of 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM, either a virus control or a cell control. [Figure 20B] Figure 20B is a bar graph showing the infection percentage of SARS-CoV-2 infected normal human bronchial epithelial (NHBE) cells treated at the time of infection with remdesivir or a viral control at concentrations of 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM (from left to right). [Figure 20C] Figure 20C is a bar graph showing (from left to right) PFU / ml of SARS-CoV-2-infected normal human bronchial epithelial (NHBE) cells treated at the time of infection with probenecid at concentrations of 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.0001 μM, or 0.00001 μM, for viral or cellular controls. [Figure 20D] Figure 20D is a bar graph showing the infection percentage of SARS-CoV-2 infected normal human bronchial epithelial (NHBE) cells treated at the time of infection with probenecid or viral control at concentrations of 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM (from left to right). [Figure 20E] Figure 20E is a bar graph (from left to right) showing the PFU / ml of SARS-CoV-2-infected normal human bronchial epithelial (NHBE) cells treated at the time of infection with 0.1 μM probenecid + 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM remdesivir, either as a virus control or a cell control. [Figure 20F]Figure 20F is a bar graph showing the infection percentage of SARS-CoV-2 infected normal human bronchial epithelial (NHBE) cells treated at the time of infection with 0.1 μM probenecid + 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM remdesivir or viral control (from left to right).

[0042] [Figure 21] Figure 21 is a bar graph showing the number of plaques in SARS-CoV-2 B.1.1.7-infected Vero cells treated at the time of infection with probenecid at concentrations of 100 μM, 50 μM, 25 μM, 12 μM, 6 μM, 3 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0.0001 μM, or 0.00001 μM, either a viral control or a cellular control. [Modes for carrying out the invention]

[0043] Detailed description of the invention I. Definition As used herein, the terms “individual,” “host,” “subject,” and “patient” are interchangeable herein and refer to animals, in particular birds and mammals, including but not limited to primates, e.g., humans, bats, rodents, e.g., mice and rats, and other laboratory animals.

[0044] As used herein, the terms “effective dose” or “therapeutic effective dose” mean a dose sufficient to treat, inhibit or alleviate one or more symptoms of a medical condition being treated, or otherwise provide the desired pharmacological and / or physiological effect. The exact dose will vary depending on various factors, such as subject-dependent variables (e.g., age, immune system health), the disease, and the treatment being administered.

[0045] As used herein, the terms “carrier” or “excipient” refer to an organic or inorganic component, a natural or synthetic non-active ingredient, in a formulation that combines one or more active ingredients.

[0046] As used herein, the term “pharmaceutically acceptable” means a non-toxic substance that does not interfere with the efficacy of the biological activity of the active ingredient.

[0047] As used herein, the term “treatment” refers to the medical management of a patient with the intention of curing, improving, stabilizing or preventing a disease, pathological condition, or disorder. This term includes active treatment, i.e., treatment explicitly directed toward improvement of a disease, pathological condition, or disorder, and also causal treatment, i.e., treatment directed toward the removal of the cause of the associated disease, pathological condition, or disorder. Furthermore, this term includes palliative treatment, i.e., treatment designed to alleviate symptoms rather than cure a disease, pathological condition, or disorder; preventive treatment, i.e., treatment directed toward minimizing, or partially or completely suppressing, the onset of the associated disease, pathological condition, or disorder; and supportive treatment, i.e., treatment used to complement another specific treatment directed toward improvement of the associated disease, pathological condition, or disorder.

[0048] The enumeration of value ranges in this specification is intended solely as a convenient way to refer to each individual value within the range individually, unless otherwise indicated herein, and each individual value is incorporated herein as if it were individually enumerated herein.

[0049] The use of the term "approximately" is intended to describe a value that is either above or below the stated value within a range of approximately + / - 10%, and in other forms, the value may be a range of either above or below the stated value within a range of approximately + / - 5%, and in other forms, the value may be a range of either above or below the stated value within a range of approximately + / - 2%, and in other forms, the value may be a range of either above or below the stated value within a range of approximately + / - 1%. The aforementioned ranges are intended to be clear from the context and do not imply further limitation.

[0050] Materials, compositions, and components that can be used for, in conjunction with, or in preparation thereof, or are products thereof are disclosed. These and other materials are disclosed herein, and where combinations, subsets, interactions, groups, etc., of these materials are disclosed, specific references to various individual and collective combinations and substitutions of these compounds are not explicitly disclosed, but it is understood that each is specifically intended and described herein. For example, where ligands are disclosed and discussed, and many substitutions that can be made for many molecules containing ligands are discussed, all kinds of combinations and substitutions of ligands and possible modifications are specifically intended unless otherwise indicated. Thus, where classes A, B, and C and classes D, E, and F and examples of combined molecules A-D are disclosed, each is intended individually and collectively, even if each is not individually enumerated. Therefore, in this example, each of the combinations A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F should be considered to be specifically contemplated and disclosed from the disclosures of A, B, and C;D, E, and F; and combination examples A-D. Similarly, any subset or combination of these is also specifically contemplated and disclosed. Therefore, for example, the subgroups A-E, B-F, and C-E should be considered to be specifically contemplated and disclosed from the disclosures of A, B, and C;D, E, and F; and combination examples A-D. Furthermore, each of the materials, compositions, components, etc., contemplated and disclosed as described above may be specifically and independently included in or excluded from any group, subgroup, list, set, etc., of such materials.

[0051] These concepts apply to all aspects of this application and include, but are not limited to, steps in a method for producing and using the disclosed compositions. Therefore, where there are various additional steps that can be performed, each of these additional steps may be performed in any particular embodiment or combination of embodiments of the disclosed method, and each such combination should be considered to be specifically contemplated and disclosed.

[0052] All methods described herein may be performed in any suitable order unless otherwise indicated or unless it is clearly inconsistent with the context. Any examples or exemplary language provided herein (e.g., "etc.") are intended merely to further illustrate the embodiments and do not limit the scope of the embodiments unless specifically claimed. No language herein should be construed as indicating that any unclaimed element is essential for the practice of the invention.

[0053] II, Composition A. Probenecid, metabolites, analogues, and pharmaceutically acceptable salts thereof The disclosed method involves administering to a subject in need of administration an effective amount of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof (including, but not limited to, its sodium salt).

[0054] Probeneside (4-[(dipropylamino)sulfonyl-1]benzoic acid (CAS number 57-66-9)) has the following structure: [ka] It possesses and is sold under the product names BENEMID® and PROBALAN®.

[0055] Probenecid is a highly lipid-soluble benzoic acid derivative with an excellent safety profile, developed in the 1950s to reduce the renal tubular excretion of penicillin. Probenecid (USP) is a white or nearly white fine crystalline powder. Probenecid is soluble in dilute alkalis, alcohols, chloroform, and acetone. It is substantially insoluble in water and dilute acids. It has a half-life of 6–12 hours. See also Drugbank accession number DB01032 (APRD00167) and PubChem CID 4911.

[0056] Metabolites and analogues of probenecid are known, for example, Guarino et al., "Mass spectral identification of probenecid metabolites in rat bile," Eur.J.Pharmacol., 8, 244-252 (1969); Perel et al., "Identification and renal excretion of probenecid metabolites in man," Life Sciences, 9, 23, 1337-1343 (1970); Perel et al., "Studies of the renal excretion of probenecid acyl glucuronide in man," Eur.J.Clin.Pharmacol, 3, 106-112 (1971); Dayton and Perel, "The metabolism of probenecid in man," NYAcad.Sci., 179, 399-402 (1971); Dayton et al., "The effect of probenecid, phenylbutazone and their analogues on the excretion of See "L-ascorbic acid in rats" J.Med.Chem.9,941-944(1966), and Israili et al., "Metabolites of probenecid. Chemical, physical, and pharmacological studies" J.Med.Chem.,15,7,709-713(1972). Each of these is specifically incorporated as a whole by reference.

[0057] In some embodiments, the metabolite is a glucuronide derivative of probenecid, such as acyl glucuronide or β-ether glucuronide.

[0058] Examples of probenecid metabolites and analogues include: dl-p-(N-propyl(Propy)-N-2-hydroxypropylsulfamoyl)benzoic acid, Propylaminopropyl acetate, Propylaminopropane-3-ol, p-(N-propyl-N-3-hydroxypropylsulfamoyl)benzoic acid, Propylaminopropionitrile, p-(N-propyl-N-3-propionnitrilosulfamoyl)benzoic acid, p-(N-propyl-N-2-carboxyethylsulfamoyl)benzoic acid, p-(N-propylsulfamoyl)benzoic acid, p-(N,N-pentamethylenesulfamoyl)benzoic acid (piperidyl analog), p-(N-propyl-N-2-propenylsulfamoyl)benzoic acid, and p-(N-propyl-N-2-oxopropylsulfamoyl)benzoic acid These include, but are not limited to, the following:

[0059] Typically, the metabolites or analogues can treat coronaviruses when administered alone or in effective amounts as discussed herein, in conjunction with their further metabolism by a subject. For example, in some embodiments, the metabolites or analogues can reduce viral replication, either alone or in conjunction with their further metabolism by a subject.

[0060] B. Formulations Probenecid, its metabolites and analogues, and pharmaceutically acceptable salts thereof can be formulated into pharmaceutical compositions. These pharmaceutical compositions may be administered via parenteral (intramuscular, intraperitoneal, intravenous (IV), or subcutaneous injection), enteral, transdermal (passively, or using iontophoresis or electroporation), or transmucosal (nasal, lung, vaginal, rectal, or sublingual) routes, or using bioerosive inserts, and may be formulated into dosage forms suitable for each route of administration.

[0061] The composition can be administered systemically.

[0062] Compositions can be formulated for immediate release, sustained release, or modified release. Delayed-release formulations are formulations that release a drug (or more drugs) at a time other than immediately after administration. Sustained-release formulations are formulations that allow for at least a twofold reduction in the frequency of administration compared to drugs presented as conventional formulations (e.g., as a liquid or as a conventional solid formulation that facilitates drug release). Modified-release formulations are selected so that the drug release characteristics over time and / or at location achieve therapeutic or convenience purposes not provided by conventional formulations such as liquids, ointments, or rapidly dissolving formulations. Delayed-release and sustained-release formulations, as well as combinations thereof, are types of modified-release formulations.

[0063] The formulation is prepared using a pharmaceutically acceptable “carrier” composed of materials that are considered safe and effective and can be administered to an individual without causing undesirable biological side effects or interactions. The “carrier” is all components present in the pharmaceutical formulation other than one or more active ingredients. The term “carrier” includes, but is not limited to, diluents, binders, lubricants, disintegrants, fillers, and coating compositions.

[0064] The "carrier" also includes all components of the coating composition, which may include plasticizers, pigments, colorants, stabilizers, and flow enhancers. Delayed-release formulations can be prepared as described in references such as "Pharmaceutical dosage forms: tablets" Liberman et al. (eds.), (New York, Marcel Dekker, Inc., 1989), "Remington - The science and practice of pharmacy," 21st edition, Lippincott Williams & Wilkins, Baltimore, MD, 2006, and "Ansel's Pharmaceutical dosage forms and drug delivery systems," 11th edition, Loyd Allen., (Media, PA: Williams and Wilkins, 2017), which provide information on carriers, materials, apparatus, and processes for preparing tablets and capsules as well as delayed-release dosage forms of tablets, capsules, and granules.

[0065] Compounds can be administered to a target with or without a delivery vehicle. Suitable delivery vehicles for compounds are known in the art and can be selected to suit a particular active agent. For example, in some embodiments, the active agent(s) are incorporated into, encapsulated by, or bound to nanoparticles, microparticles, micelles, synthetic lipoprotein particles, or carbon nanotubes. For example, a composition can be incorporated into a vehicle such as polymer microparticles that provide controlled release of the active agent(s). In some embodiments, the release of the drug(s) is controlled by diffusion of the active agent(s) from the microparticles and / or degradation of the polymer particles by hydrolysis and / or enzymatic degradation.

[0066] Suitable polymers include ethylcellulose and other natural or synthetic cellulose derivatives. Polymers that dissolve slowly and form gels in aqueous environments, such as hydroxypropyl methylcellulose or polyethylene oxide, may also be suitable as materials for drug-containing microparticles or particles. Other polymers include, but are not limited to, polyacid anhydrides, poly(esteric acid anhydrides), polyhydroxy acids, such as polylactide (PLA), polyglycolide (PGA), poly(lactide-co-glycolide) (PLGA), poly-3-hydroxybutyrate (PHB) and its copolymers, poly-4-hydroxybutyrate (P4HB) and its copolymers, polycaprolactone and its copolymers, and combinations thereof. In some embodiments, both drugs are incorporated into the same particle and formulated to be released over different times and / or periods. For example, in some embodiments, one of the drugs is completely released from the particle before the release of the second drug begins. In other embodiments, the release of the first drug is initiated, followed by the release of the second drug before all of the first drug has been released. In yet another embodiment, both drugs are released simultaneously over the same period or over different periods.

[0067] 1. Oral immediate-release formulation Suitable oral dosage forms include tablets, capsules, liquids, suspensions, syrups, and lozenges. Therefore, compositions can be formulated as solids or liquids. Tablets can be made using compression or molding techniques well known in the art. Gelatin or non-gelatin capsules can be prepared as rigid or flexible capsule shells capable of encapsulating liquid, solid, and semi-solid filler materials using techniques well known in the art.

[0068] Examples of suitable coating materials include, but are not limited to, cellulose polymers such as cellulose phthalate acetate, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and hydroxypropyl methylcellulose succinate acetate; polyvinyl phthalate acetate, acrylic polymers and copolymers, as well as methacrylic resins, zein, shellac, and polysaccharides, which are commercially available under the trade name Eudragit® (Roth Pharma, Westerstadt, Germany).

[0069] Furthermore, the coating material may contain conventional carriers such as plasticizers, pigments, colorants, flow promoters, stabilizers, pore-forming agents, and surfactants.

[0070] Pharmaceutically acceptable excipients present in drug-containing tablets, beads, granules, or particles as needed include, but are not limited to, diluents, binders, lubricants, disintegrants, colorants, stabilizers, and surfactants. Diluents, also called “fillers,” are typically required to increase the bulk of solid dosage forms so that a practical size is provided for the compression of tablets or the formation of beads and granules. Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolyzed starch, pregelatinized starch, silicone dioxide, titanium dioxide, magnesium aluminum silicate, and powdered sugar.

[0071] Binders are used to impart tackiness to solid dosage forms, thus ensuring that tablets, beads, or granules remain intact after the formation of the dosage form. Suitable binder materials include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (including sucrose, glucose, dextrose, lactose, and sorbitol), polyethylene glycol, waxes, natural and synthetic rubbers, such as acacia, tragacanth, sodium alginate, cellulose including hydroxypropyl methylcellulose, hydroxypropylcellulose, and ethylcellulose, veegum, and synthetic polymers, such as acrylic acid and methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid / polymethacrylic acid, and polyvinylpyrrolidone.

[0072] Lubricants are used to facilitate tablet manufacturing. Suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, polyethylene glycol, talc, and mineral oil.

[0073] Disintegrants are used to facilitate the breakdown or "breakup" of the dosage form after administration and commonly include, but are not limited to, starch, sodium starch glycolate, sodium carboxymethyl starch, sodium carboxymethylcellulose, hydroxypropylcellulose, pregelatinized starch, clay, cellulose, arginine, gum, or cross-linked polymers, such as cross-linked PVP (Polyplasdone XL from GAF Chemical Corp).

[0074] Stabilizers are used, for example, to inhibit or delay drug degradation reactions, including oxidation reactions.

[0075] Surfactants can be anionic, cationic, amphoteric, or nonionic surfactants. Suitable anionic surfactants include, but are not limited to, those containing carboxylate ions, sulfonate ions, and sulfate ions. Examples of anionic surfactants include long-chain alkyl sulfonates such as sodium, potassium, ammonium, and alkylaryl sulfonates such as sodium dodecylbenzenesulfonate; sodium dialkyl sulfosuccinate, e.g., sodium dodecylbenzenesulfonate; sodium dialkyl sulfosuccinate, e.g., sodium bis-(2-ethylthioxyl)-sulfosuccinate; and alkyl sulfates, e.g., sodium lauryl sulfate. Cationic surfactants include, but are not limited to, quaternary ammonium compounds such as benzalkonium chloride, benzethonium chloride, cetrimonium bromide, stearyldimethylbenzylammonium chloride, polyoxyethylene, and coconutamines. Examples of nonionic surfactants include ethylene glycol monostearate, propylene glycol myristate, glyceryl monostearate, glyceryl stearate, polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150 laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbate, polyoxyethylene octylphenyl ether, PEG-1000 cetyl ether, polyoxyethylene tridecyl ether, polypropylene glycol butyl ether, POLOXAMER® 401, stearoyl monoisopropanolamide, and polyoxyethylene hydrogenated tallow amide. Examples of amphoteric surfactants include sodium N-dodecyl-β-alanine, sodium N-lauryl-β-iminodipropionate, myristoamphoacetate, lauryl betaine, and lauryl sulfobetaine.

[0076] If desired, tablets, bead granules, or particles may also contain small amounts of non-toxic auxiliary substances such as wetting agents or emulsifiers, dyes, pH buffers, and preservatives.

[0077] 2. Sustained-release dosage form Sustained-release formulations are generally prepared as diffusion or permeation systems, as described, for example, in "Remington—The science and practice of pharmacy" (21st edition, Lippincott Williams & Wilkins, Baltimore, MD, 2006). Diffusion systems typically consist of two types of devices: a reservoir and a matrix, which are well known and described in the art. Matrix devices are generally prepared by compressing a drug into tablet form with a polymer carrier that slowly dissolves the drug. Three main types of materials used in the preparation of matrix devices are insoluble plastics, hydrophilic polymers, and alipid compounds. Examples of plastic matrices include, but are not limited to, methyl acrylate-methyl methacrylate, polyvinyl chloride, and polyethylene. Examples of hydrophilic polymers include, but are not limited to, methylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and carbopol 934, and polyethylene oxide. Examples of alipid compounds include, but are not limited to, various waxes such as carnauba wax and glyceryl tristearate.

[0078] Alternatively, sustained-release formulations can be prepared using an infiltration system or by applying a semipermeable coating to the dosage form. In the latter case, the desired drug release profile can be achieved by combining low-permeability and high-permeability coating materials in appropriate proportions.

[0079] Devices with the different drug release mechanisms described above can be combined into a final dosage form containing one or more units. Examples of multiple units include multilayer tablets, tablets, beads, and capsules containing granules.

[0080] The immediate-release portion can be added to the sustained-release system by applying an immediate-release layer on top of the sustained-release core using a coating or compression process, or in a multi-unit system such as a capsule containing sustained-release and immediate-release beads.

[0081] Sustained-release tablets containing hydrophilic polymers are prepared by techniques commonly known in the art, such as direct compression, wet granulation, or dry granulation processes. These formulations typically incorporate polymers, diluents, binders, lubricants, and active pharmaceutical ingredients. Common diluents include inert powders such as any of many different types of starch, powdered cellulose, particularly crystalline and microcrystalline cellulose, sugars such as fructose, mannitol, and sucrose, grain flour, and similar edible powders. Typical diluents include, for example, various types of starch, inorganic salts such as lactose, mannitol, kaolin, calcium phosphate or sulfate, and sodium chloride, and powdered sugars. Powdered cellulose derivatives are also useful. Typical tablet binders include substances such as starch and gelatin, as well as sugars such as lactose, fructose, and glucose. Natural and synthetic rubbers, including acacia, alginates, methylcellulose, and polyvinylpyrrolidine can also be used. Polyethylene glycol, hydrophilic polymers, ethylcellulose, and waxes can also serve as binders. To prevent tablets and punches from sticking to the die, a lubricant is required in tablet formulations. The lubricant is selected from slippery solids such as talc, magnesium stearate and calcium stearate, stearic acid, and hydrogenated vegetable oils.

[0082] Sustained-release tablets containing wax materials are generally prepared using methods known in the art, such as direct blending, coagulation, and aqueous dispersion. In the freezing method, the drug is mixed with the wax material and then spray-coagulated or coagulated and sieved.

[0083] 3. Delayed-release dosage form Delayed-release formulations are prepared by coating a solid dosage form with a polymer film that is insoluble in the acidic environment of the stomach and soluble in the neutral environment of the small intestine.

[0084] Delayed-release dosage units can be prepared, for example, by coating a drug or drug-containing composition with a selected coating material. The drug-containing composition may be, for example, a tablet for incorporation into a capsule, a tablet for use as an inner core of a "coated core" dosage form, or a plurality of drug-containing beads, particles, or granules for incorporation into either a tablet or a capsule. Preferred coating materials include bioerosive, gradually hydrolyzable, slowly soluble in water, and / or enzymatically degradable polymers, which may be conventional "enteric-coated" polymers. Enteric-coated polymers, as will be understood by those skilled in the art, become soluble or slowly eroded in the higher pH environment of the lower gastrointestinal tract as the dosage form passes through the gastrointestinal tract, while enzymatically degradable polymers are broken down by bacterial enzymes present in the lower gastrointestinal tract, particularly the colon.Suitable coating materials for delayed release include cellulose polymers, such as hydroxypropylcellulose, hydroxyethylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose acetate succinate, hydroxypropylmethylcellulose phthalate, methylcellulose, ethylcellulose, cellulose acetate, cellulose acetate phthalate, cellulose acetate trimellitate, and sodium carboxymethylcellulose; preferably acrylic acid, methacrylic acid, methyl acrylate, ethylcellulose; The trade name EUDRAGIT® (Rohm) includes acrylate, methyl methacrylate and / or ethyl methacrylate, as well as EUDRAGIT® L30D-55 and L100-55 (soluble at pH 5.5 and above), EUDRAGIT® L-100 (soluble at pH 6.0 and above), EUDRAGIT® S (soluble at pH 7.0 and above as a result of a relatively high degree of esterification), and EUDRAGIT® NE, RL and RS (water-insoluble polymers with different degrees of permeability and swelling). Examples include, but are not limited to, acrylic polymers and copolymers formed from other methacrylic resins commercially available in Pharma (Westerstadt, Germany); vinyl polymers and copolymers, such as polyvinylpyrrolidone, vinyl acetate, vinyl phthalate acetate, vinyl acetate crotonic acid copolymer and ethylene vinyl acetate copolymer; enzymatically digestible polymers, such as azopolymers, pectin, chitosan, amylose and guar gum; zein and shellac. Combinations of different coating materials may be used. Multilayer coatings using different polymers may be applied.

[0085] The preferred coating weight of a particular coating material can be easily determined by those skilled in the art by evaluating the individual release profiles of tablets, beads, and granules prepared with different amounts of various coating materials. The combination of materials, methods, and application modes that yields the desired release characteristics can only be determined from clinical studies.

[0086] The coating composition may contain conventional additives such as plasticizers, pigments, colorants, stabilizers, and flow promoters. Plasticizers are typically present to reduce the brittleness of the coating and generally amount to about 10% to 50% by weight relative to the dry weight of the polymer. Typical examples of plasticizers include polyethylene glycol, propylene glycol, triacetin, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethyl citrate, tributyl citrate, triethyl acetyl citrate, castor oil, and acetylated monoglycerides. It is preferable to use stabilizers to stabilize the particles in the dispersion. Typical stabilizers are nonionic emulsifiers such as sorbitan esters, polysorbates, and polyvinylpyrrolidone. Flow promoters are recommended to reduce tackiness during film formation and drying and generally amount to about 25% to 100% by weight of the polymer in the coating solution. One effective flow promoter is talc. Other flow promoters such as magnesium stearate and glycerol monostearate may also be used. Pigments such as titanium dioxide may be used. A small amount of an antifoaming agent, such as silicone (e.g., simethicone), may be added to the coating composition.

[0087] Manufacturing method As will be understood by those skilled in the art, and as described in relevant texts and literature, many methods are available for preparing drug-containing tablets, beads, granules, or particles that provide a variety of drug release profiles. Such methods include, but are not limited to, coating a drug or drug-containing composition with a suitable coating material, typically not necessarily incorporating polymer materials, increasing the drug particle size, locating the drug within a matrix, and forming a complex of the drug with a suitable complexing agent.

[0088] Delayed-release dosage units can be coated with a delayed-release polymer coating using conventional techniques, such as conventional coating pans, airless spray technology, or fluidized bed coating equipment (with or without Wurster inserts). For detailed information on materials, equipment, and processes for preparing tablets and delayed-release dosage forms, see Pharmaceutical Dosage Forms: Tablets, Lieberman et al. (eds.), (New York: Marcel Dekker, Inc., 1989) and Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 11th edition (Media, PA: Williams & Wilkins, 2017).

[0089] A preferred method for preparing sustained-release tablets is by compressing a drug-containing blend, such as a blend of granules, prepared using direct blending, wet granulation, or dry granulation processes. Sustained-release tablets may also be molded from a wet material containing a suitable water-soluble lubricant, rather than being compressed. However, tablets are preferably manufactured using compression rather than molding. A preferred method for forming a sustained-release drug-containing blend is to directly mix drug particles with one or more excipients, such as diluents (or fillers), binders, disintegrants, lubricants, flow enhancers, and colorants. As an alternative to direct blending, drug-containing blends may be prepared by using wet granulation or dry granulation processes. Beads containing active agents can also be prepared by one of a number of prior arts, typically starting from dispersions. For example, a typical method for preparing drug-containing beads involves dispersing or dissolving an active agent in a coating suspension or solution containing pharmaceutical excipients such as polyvinylpyrrolidone, methylcellulose, talc, metal stearate, silicone dioxide, and plasticizers. The mixture is used to coat bead cores, such as spheroidal sugars (or "nonpareils"), that have a size of approximately 60-20 mesh.

[0090] An alternative procedure for preparing drug beads involves blending the drug with one or more pharmaceutically acceptable excipients such as microcrystalline cellulose, lactose, cellulose, polyvinylpyrrolidone, talc, magnesium stearate, and disintegrants; extruding this blend; spheroidizing the extruded material; drying it; and coating it as needed to form immediate-release beads.

[0091] 4. Formulations for mucosal and pulmonary administration Probenecid, its metabolites and analogues, and its pharmaceutical compositions can be formulated for pulmonary or mucosal administration. Administration may involve delivery of the composition to the lungs, nose, oral cavity (sublingual, buccal), vaginal, or rectal mucosa. In certain embodiments, the composition is formulated for a subject and delivered to the subject sublingually.

[0092] In some embodiments, compounds are formulated for pulmonary delivery, such as intranasal administration or oral inhalation. The airways are structures involved in the exchange of gases between the atmosphere and the bloodstream. The lungs are branched structures that ultimately end in the alveoli where gas exchange takes place. The alveolar surface area is the largest in the respiratory system and is where drug absorption occurs. Alveoli are covered by a thin epithelium without ciliary or mucous layers and secrete surfactant phospholipids. The airways include the upper airways, which include the oropharynx and larynx, followed by the lower airways, which include the trachea, and then branching into the bronchi and bronchioles. The upper and lower airways are called the inductive airways. The terminal bronchioles then branch into respiratory bronchioles, leading to the alveoli or deep lungs, which are the final respiratory region. The deep lungs or alveoli are the primary targets of inhalation therapeutic aerosols for systemic drug delivery.

[0093] Lung administration of therapeutic compositions consisting of low molecular weight drugs, such as β-androgen antagonists used to treat asthma, has been observed. Other pulmonary-active therapeutic agents are administered systemically and targeted via pulmonary absorption.

[0094] Nasal delivery is considered a promising technique for administering therapeutic drugs for the following reasons: the nose has a large surface area available for drug absorption due to the covering of the epithelial surface by numerous microvilli; the subepithelial layer is highly angiogenic; venous blood from the nose enters the systemic circulation directly, thus avoiding drug loss due to first-pass metabolism in the liver; lower doses; faster achievement of therapeutic blood levels; faster onset of pharmacological activity; fewer side effects; 1 cm 3 It provides high total blood flow, a porous endothelial basement membrane, and is easily accessible.

[0095] In some embodiments, the composition is formulated as an aerosol. As used herein, the term aerosol refers to any preparation of a fine mist of particles, which may be a solution or suspension, whether or not it is produced using a propellant. Aerosols can be produced using standard techniques such as sonication or high-pressure processing.

[0096] Probenecid is used to mitigate the nephrotoxicity of cidofovir when administered intravenously. Direct delivery of cidofovir to the airways has also been shown to be an effective prophylactic strategy that maximizes tissue concentration at the initial viral replication site while minimizing its accumulation in the kidneys (Roy et al., Antimicrob Agents Chemother., 47(9):2933-2937 (2003)).

[0097] Carriers for lung formulations can be divided into those for dry powder formulations and those for administration as solutions. Aerosols for delivering therapeutic drugs to the airways are known in the art. For administration via the upper respiratory tract, formulations may be formulated as liquids, e.g., water or isotonic saline (buffered or unbuffered), or as suspensions; for intranasal administration, they may be formulated as drops or sprays. Preferably, such liquids or suspensions are isotonic with respect to nasal secretions, for example, with approximately the same pH in the range of about pH 4.0 to about pH 7.4 or pH 6.0 to pH 7.0. Buffers should be physiologically compatible and include, simply as an example, phosphate buffers. For example, typical decongestants are described as being buffered to a pH of about 6.2. Those skilled in the art can easily determine the appropriate saline content and pH for a harmless aqueous solution for intranasal and / or upper respiratory tract administration.

[0098] Preferably, the aqueous solution is a physiologically acceptable aqueous solution containing water, a salt and / or a buffer, such as phosphate-buffered saline (PBS), or any other aqueous solution acceptable for administration to animals or humans. Such solutions are well known to those skilled in the art and include, but are not limited to, distilled water, deionized water, pure water or ultrapure water, saline solution, and phosphate-buffered saline (PBS). Other suitable aqueous vehicles include, but are not limited to, Ringer's solution and isotonic sodium chloride. Suitable aqueous suspensions include suspending agents such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone and tragacanth gum, and wetting agents such as lecithin. Suitable preservatives for aqueous suspensions include ethyl p-hydroxybenzoate and n-propyl p-hydroxybenzoate.

[0099] In another embodiment, solvents that are low-toxicity organic (i.e., non-aqueous) Class 3 residual solvents, such as ethanol, acetone, ethyl acetate, tetrahydrofuran, ethyl ether, and propanol, can be used in the formulation. The solvent is selected based on its ability to readily aerosolize the formulation. The solvent must not react toxicly with the compound. A suitable solvent should be used that dissolves the compound or forms a suspension of the compound. The solvent must be sufficiently volatile to allow the formation of an aerosol of the solution or suspension. Additional solvents or aerosolizers, such as Freon, can be added as desired to increase the volatility of the solution or suspension.

[0100] In one embodiment, the composition may contain small amounts of polymers, surfactants, or other excipients well known to those skilled in the art. In this context, “small amounts” means that there are no excipients that affect or mediate the uptake of the compound in the lungs, and that any excipients present are present in amounts that do not adversely affect the uptake of the compound in the lungs.

[0101] Dry lipid powders can be directly dispersed in ethanol due to their hydrophobic properties. For lipids stored in organic solvents such as chloroform, the desired amount of solution is placed in a vial, and the chloroform is evaporated under a nitrogen stream to form a dry thin film on the surface of the glass vial. Upon reconstitution with ethanol, the film swells readily. To completely disperse the lipid molecules in the organic solvent, the suspension is sonicated. Non-aqueous lipid suspensions can also be prepared in anhydrous ethanol using a reusable PARI LC Jet+ nebulizer (PARI Respiratory Equipment, Monterey, CA).

[0102] Dry powder formulations with large particle sizes ("DPFs") exhibit improved flow properties, including less aggregation, easier aerosolization, and potentially lower phagocytosis. Dry powder aerosols for inhalation therapy are generally manufactured with an average diameter primarily in the range of less than 5 microns, although the preferred range is 1 to 10 microns in aerodynamic diameter. Among other possible advantages, large "carrier" particles (without the drug) are co-delivered with the therapeutic aerosol to help achieve efficient aerosolization.

[0103] Polymer particles can be prepared using single and double emulsion solvent evaporation, spray drying, solvent extraction, solvent evaporation, phase separation, simple and complex coacervation, interfacial polymerization, and other methods well known to those skilled in the art. The particles can also be prepared using methods known in the art for producing microspheres or microcapsules. Preferred production methods are by spray drying and freeze-drying, which involve using a solution containing a surfactant, spraying it to form droplets of the desired size, and removing the solvent.

[0104] Particles can be manufactured with appropriate materials, surface roughness, diameter, and tap density for local delivery to selected areas of the airways, such as the deep lungs or upper airways. For example, higher density or larger particles can be used for upper airway delivery. Similarly, mixtures of particles of different sizes with the same or different active agents can be administered in a single dose to target different areas of the lungs.

[0105] A composition and method for preparing an inhalable pharmaceutical composition containing probenecid is described in U.S. Patent Application Publication No. 2015 / 0272870.

[0106] Accordingly, formulations and methods are provided for administering the disclosed compositions to the nasal mucosa and / or lungs by intranasal delivery and to the lungs by oral inhalation. With respect to intranasal delivery, formulations and delivery devices can be selected and prepared to facilitate absorption via the nasal mucosa or lungs. The nasal mucosa is more readily accessible than other mucous membranes and provides a practical entry portal for small and large molecules (Bitter et al., "Nasal Drug Delivery in Humans," in Surber et al. (eds.): Topical Applications and the Mucosa. Curr Probl Dermatol. Basel, Karger, 2011, vol 40, pp 20-35; Pires et al., J Pharm Pharmaceut Sci., 12(3)288-311, 2009; and Djupesland, Drug Deliv. and Transl. Res., 3:42-62 (2013) DOI 10.1007 / s13346-012-0108-9). Intranasal administration offers rapid onset of therapeutic effect, reduced first-pass effect, decreased gastrointestinal degradation and pulmonary toxicity, non-invasiveness, essentially painless application, and easy and immediate use by patients (especially suitable for children) or by physicians in emergency settings. For example, Flu Mist® is an exemplary effective intranasal influenza vaccine spray.

[0107] Numerous delivery devices are available for intranasal administration. These devices differ in delivery accuracy, dose reproducibility, cost, and ease of use. Metered-dose systems offer dose accuracy and reproducibility. Differences also exist in delivery force, spray pattern, and droplet size. The latter are crucial for intranasal drug deposition. Parameters can be modulated to promote deposition while limiting the percentage of small particles that can bypass the nose and enter the lungs, or to decrease deposition while increasing the percentage of small particles that can bypass the nose and enter the lungs.

[0108] The following aspects of the nasal anatomical structure can affect drug delivery. During exhalation, the soft palate automatically closes, separating the nasal cavity from the oral cavity. This allows for the use of relatively small particles in nasal sprays and further avoids lung deposition. Furthermore, during soft palate closure, there is a connecting passage between the two nostrils located behind the wall separating the two passages. Under these circumstances, airflow can enter through one nostril and exit through the other. This bidirectional delivery concept integrates two anatomical facts into one fully functional device. The device is inserted into one nostril by a sealing nozzle, and the patient blows into the mouthpiece. The combination of the closed soft palate and the sealing nozzle generates an airflow that enters one nostril, swirls 180° through the connecting passage, and exits through the other nostril (bidirectional flow). Because delivery occurs during exhalation, small particles cannot enter the lungs.

[0109] For efficient delivery to the nasal mucosa, particle size, flow rate, and direction can be adjusted. Further control of input pressure by adding an outlet resistor can improve distribution to the sinuses and middle ear. Manipulation of the flow pattern allows delivery to the olfactory region, thereby achieving, in some cases, direct "nose-to-brain" delivery. A 180-degree swirl behind the septum captures still-floating particles, enabling targeted delivery of cargo to the adenoids.

[0110] Strategies for enhancing drug absorption via nasal and pulmonary routes are also known in the art and can be utilized in disclosed formulations and delivery methods. Such strategies include, for example, the use of absorption enhancers such as surfactants, cyclodextrins, protease inhibitors, and tight junction modulators, as well as the application of carriers such as liposomes and nanoparticles. See, for example, Ghadiri et al., Pharmaceutics, 11(3):113 (2019).

[0111] 5. Parenteral administration formulations Probenecid, its metabolites and analogues, and its pharmaceutical compositions can be administered in aqueous solution by parenteral injection or infusion. Formulations may also be in the form of suspensions or emulsions. Generally, pharmaceutical compositions comprising an effective amount(s) of the active agent(s) are provided and optionally include pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and / or carriers. Such compositions may include diluents, sterile water, the contents of various buffers (e.g., Tris-HCl, acetates, phosphates), buffered salines of pH and ionic strength; and optionally additives, such as detergents and solubilizers (e.g., TWEEN® 20, TWEEN® 80, also known as POLYSORBATE® 20 or 80), antioxidants (e.g., ascorbic acid, sodium metabisulfite), and preservatives (e.g., thimerosal, benzyl alcohol) and fillers (e.g., lactose, mannitol). Examples of non-aqueous solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. The formulation may be lyophilized and redissolved / resuspended immediately before use. The formulation may be sterilized, for example, by filtration through a bacterial-retaining filter, by incorporating a sterilizing agent into the composition, by irradiation of the composition, or by heating the composition.

[0112] III. Treatment Methods The present invention provides methods for treating viral infections in subjects requiring treatment for viral infections. In some embodiments, the virus may be a virus that causes respiratory illness or disease. Therefore, methods for treating respiratory illness or disease in subjects infected with a virus are also provided.

[0113] This method may involve administering an effective amount of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof, to a subject to reduce viral replication, infection, or a combination thereof. In some embodiments, the amount is effective in reducing viral titer in the subject, reducing the host cell assembly of the virus, reducing and / or limiting the excess inflammation associated with infection, and / or reducing and / or limiting one or more severe respiratory symptoms.

[0114] In some embodiments, the subject is exposed to or will be exposed to the virus. In some embodiments, the subject is exposed to the virus or is experiencing an active viral infection.

[0115] In some embodiments, viral infection is detected by PCR tests designed to detect viral RNA in a sample from the subject, such as a nasal swab, throat swab, saliva, or other bodily fluids, or by serological or immunodiagnostic tests designed to detect antibodies produced by the body's immune system in response to the infection, typically in a blood sample from the subject.

[0116] The composition can also be administered prophylactically, for example, to reduce or prevent the effects of future exposure to the virus and any associated infection. Therefore, in some embodiments, the subject has not been exposed to the virus and / or has not yet experienced an active viral infection. In some embodiments, the subject is a healthy subject.

[0117] In some embodiments, subjects had close contact with subjects who had tested positive for the virus. Such subjects may or may not exhibit one or more symptoms of the infection. Close contact may include, for example, being within 2 meters of an infected person for a total of 15 minutes or more, providing home care to an infected person, having direct physical contact with an infected person (hugging or kissing), sharing dishes or drinking utensils with an infected person, being sneezed on by an infected person, being coughed on by an infected person, or otherwise coming into contact with respiratory droplets from an infected person.

[0118] In some embodiments, subjects were identified in contact tracing as one or more subjects who had been exposed to or infected with the virus.

[0119] In some embodiments, the subjects are exposed to the virus. An exemplary subject is a healthcare worker treating an infected person.

[0120] In some embodiments, the treatment is initiated one, two, three, four, or five hours or more before or after exposure to the virus, or several days or weeks before or after.

[0121] In some embodiments, probenecid, its metabolites or analogues, or pharmaceutically acceptable salts thereof are administered in amounts effective to alleviate or prevent one or more symptoms of a viral infection. Symptoms include those of acute respiratory illness, such as fever, sinus and / or pulmonary congestion, runny or nasal congestion, cough, sneezing, sore throat, body aches, fatigue, shortness of breath, chest tightness, and wheezing during exhalation. Specific symptoms associated with exemplary viruses and infections are discussed in more detail below. Most typically, the virus is a coronavirus.

[0122] In some embodiments, subjects do not have gout, do not require long-term serum levels of penicillin (or other antibiotics), and do not have pelvic inflammatory disease or gonorrhea.

[0123] In some embodiments, the subjects have an influenza infection. See, for example, Perwitasari et al., Antimicrob Agents Chemother, 57(1):475-83(2013).doi:10.1128 / AAC.01532-12.)). For example, in some embodiments, the subjects have an influenza (e.g., influenza A, influenza B, influenza C and / or influenza D) infection and an infection from another virus such as coronavirus. In some embodiments, the subjects do not have an influenza virus infection.

[0124] The target audience may be males and / or females, adults (e.g., 18 years of age or older) and / or children under 18 years of age.

[0125] A. Exemplary dosages and regimens Probenecid, its metabolites and analogues, and pharmaceutically acceptable salts thereof can be administered to subjects in pharmaceutical compositions such as those discussed above, and can be administered parenterally (intramuscular, intraperitoneal, intravenous (IV), or subcutaneous injection), enterally, percutaneously (passively, or using iontophoresis or electroporation), or transmucosal (nasal, lung, vaginal, rectal, or sublingual) routes, as discussed in more detail above.

[0126] The precise dosage varies depending on various factors, such as subject-dependent variables (e.g., age, immune system health, clinical symptoms, delivery route, etc.).

[0127] To treat gout, probenecid was administered orally (PO) at a dose of 250 mg twice daily for one week; the dose was increased to 500 mg PO twice daily, with a maximum increase of 500 mg to 2 g / day.

[0128] To prolong penicillin serum levels, probenecid was administered orally at a dose of 500 mg four times daily.

[0129] In cases of pelvic inflammatory disease, probenecid was administered orally as a single dose of 1 g together with 2 g of cefoxitine intramuscularly (IM).

[0130] In cases of gonorrhea, probenecid was administered orally as a single dose of 1 g together with 2 g of cefoxitine (IM).

[0131] Typically, in children (e.g., ages 2–14 years and weight less than 50 kg) as an adjuvant for antibiotic treatment, the initial dose is 25 mg / kg (or 0.7 g / m2) orally once; maintenance dose is 40 mg / kg (or 1.2 g / m2) / day orally four times a day in four equally divided doses.

[0132] Therefore, generally, as mere examples, useful dosage forms in the disclosed method may include doses in the range of 0.1 mg to 3,000 mg; 25 mg to 2,000 mg; 25 mg to 1,000 mg; 50 mg to 1,000 mg; 100 mg to 1,000 mg; or 250 mg to 1,000 mg, with exemplary doses being 10 mg, 25 mg, 45 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 750 mg, and 1,000 mg, which can be administered, for example, daily, weekly, every other week, 1, 2, 3, 4, or 5 times, over 1, 2, 3, 4 or longer weeks, and, for example, until symptoms improve or disappear. In some embodiments, a single treatment can be repeated at intervals of 1, 2, 3, 4, 5, 6, 7 or longer, such as days, weeks, or months.

[0133] In some embodiments, the treatment regimen is similar to those described above for conditions such as gout, prolonged penicillin serum levels, pelvic inflammatory disease, and gonorrhea.

[0134] In certain embodiments, probenecid or its metabolites or analogs or pharmaceutically acceptable salts thereof are administered at a dose of 250 mg twice daily.

[0135] The following results indicate that both 2 mg / kg and 200 mg / kg doses were effective in treating hamsters in vivo. Therefore, in some embodiments, the dose is between 2 mg / kg and 200 mg / kg, including both extremes.

[0136] As stated above, the enumeration of ranges of values ​​in this Specified Terms, including the dosage ranges above and elsewhere in this Specified Terms, is intended solely as a convenient way to refer individually to each individual value that falls within the range, and each individual value is incorporated herein as if it were individually enumerated herein.

[0137] The administration regimen may be, for example, intermittent or continuous (e.g., continuous infusion). The administration regimen may include administration of the same or different doses. Therefore, the administration regimen may include dose escalation, dose reduction, or a combination thereof.

[0138] In some embodiments, the composition is administered in a pulsed dosing regimen. Pulsed dosing refers to a dosing approach that brings about gradually increasing drug levels early in the dosing interval, followed by a long dose-free period. For example, in some embodiments, drug administration can be front-loaded by, for example, one, two, three, four, or five consecutive bolus doses, after which the drug level can be reduced to the next dose. In some embodiments, serum drug levels can be reduced to about zero.

[0139] This type of drug delivery technology can offer therapeutic benefits such as reduced dose frequency and improved patient compliance. Compared to intermittent dosing, pulse dosing frontloads the drug and allows for extended dose-free periods during which drug concentrations decrease to near zero. However, unlike a single high-dose bolus (e.g., administered once daily), short bursts of drug are separated by short dose-free periods, resulting in fluctuating serum concentrations (Ibrahim et al., Antimicrobial Agents and Chemotherapy, 48(11):4195-4199 (2004)). In certain embodiments, pulse dosing is performed by oral or intravenous administration. For example, in some embodiments, treatment involves short-term, discontinuous / intermittent intravenous infusions of very high doses of probenecid, its metabolites or analogues, or pharmaceutically acceptable salts thereof.

[0140] In some embodiments, a large bolus dose of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof is approximately 1,000 mg to 5,000 mg, including both ends, or any partial range or specific dose in that range.

[0141] The maximum recommended dose of probenecid is 2 grams / day (oral) for adults, adolescents, and children weighing over 50 kg, and 40 mg / kg / day (1.2 grams / m2 / day) (oral) (not exceeding 2 grams / day (oral)) for adolescents and children weighing 50 kg or less. Therefore, in some embodiments, the dose does not exceed 5 g, 4 g, 3 g, or 2 g per day. In some embodiments, the dose does not exceed 40 mg / kg / day. See also "probenecid-Drug Summary," the Prescribers' Digital Reference.

[0142] In some embodiments, the oral tablets contain, for example, 500 mg of probenecid and, optionally, one or more of the following non-active ingredients: microcrystalline cellulose, sodium lauryl sulfate, sodium starch glycolate, starch (corn), povidone, colloidal silicon dioxide, magnesium stearate, polyvinyl alcohol, titanium dioxide, polyethylene glycol, talc, D&C Yellow #10 Aluminum Lake, FD&C Yellow #6 Aluminum Lake, and FD&C Blue #2 Aluminum Lake.

[0143] In some embodiments, subjects are orally administered 500 mg of probenecid or its metabolites or analogs or pharmaceutically acceptable salts or thereof twice daily (BID) for 14 days (PO).

[0144] In some embodiments, the treatment ends 0, 1, 2, 3, 4, or 5 days after the subject's symptom(s) have disappeared, and the subject is enrolled in one, two, or more negative tests for viral infection (e.g., negative SARS-CoV-2 / COVID-19 tests), or a combination thereof.

[0145] B. Combination therapy In some embodiments, probenecid, its metabolites or analogues, or pharmaceutically acceptable salts thereof are administered in combination with one or more additional active agents. Combination therapy may involve administering the active agents together in the same mixture or in separate mixtures. Thus, in some embodiments, the pharmaceutical composition contains two, three or more active agents. Such formulations typically contain an effective amount of probenecid, its metabolites or analogues, or pharmaceutically acceptable salts thereof. In some embodiments, the second active agent is an antiviral agent (i.e., a second antiviral agent), an antipyretic, an anti-inflammatory agent, an analgesic, or a combination thereof. In certain embodiments, the antiviral agent is oseltamivir phosphate (TAMIFLU®). Tamiflu is a prescription drug used to treat flu (influenza) in people aged 2 weeks and older who have symptoms of flu. Probenecid or its metabolites, analogues, or pharmaceutical salts may enhance the effectiveness of antiviral drugs such as oseltamivir phosphate by helping to suppress drug excretion during treatment.

[0146] Some products that may interact with this drug include cancer chemotherapy, baricitinib, diffyline, ketorolac, methotrexate, pyrazinamide, salicylates (e.g., high-dose aspirin), zidovudine, and certain drugs that are eliminated from the body by the kidneys (e.g., ceftazidime / avibactam, dapsone, heparin, fosfomycin). Therefore, in some embodiments, one or more of these drugs are not administered while the subject is being treated with probenecid or its metabolites, analogues, or pharmaceutical salts.

[0147] In some embodiments, one or more additional active agents are remdesivir.

[0148] C. Exemplary viruses and symptoms Symptoms of diseases resulting from infection with exemplary viruses and viruses treatable by the disclosed methods are also provided. The viruses are typically coronaviruses. The current classification of coronaviruses recognizes 39 species in 27 subgenera, 5 genera, and 2 subfamilies, belonging to the family Coronaviridae, suborder Cornidovirineae, order Nidovirales, and kingdom Riboviria (Coronaviridae Study Group of the International Committee on Taxonomy of Viruses, Nat Microbiol 2020. DOI:10.1038 / s41564-020-0695-z). They are enveloped viruses with a positive-sense single-stranded RNA genome and a helical nucleocapsid. The genome size of coronaviruses ranges from approximately 26 to 32 kilobases, making them one of the largest among RNA viruses.

[0149] Coronaviruses cause disease in mammals and birds. Most typically, alpha- and beta-coronaviruses infect mammals, while gamma- and delta-coronaviruses primarily infect birds. At least seven of these viruses can infect humans: 229E (alpha), NL63 (alpha), OC43 (beta), HKU1 (beta), MERS-CoV (beta) virus, SARS-CoV (beta), and SARS-CoV-2 (beta).

[0150] In a preferred embodiment, the subject is human. In humans, coronaviruses can cause respiratory infections that can range from mild to fatal. Mild illnesses include some cases of the common cold, while more fatal types can cause SARS, MERS, and COVID-19 (i.e., those caused by SARS-CoV-2).

[0151] The subjects may have one or more symptoms characteristic of SARS, MERS, or COVID-19.

[0152] SARS (i.e., SARS-CoV) typically begins with flu-like signs and symptoms such as fever, chills, muscle aches, headache, and occasional diarrhea. About a week later, signs and symptoms include a fever of 100.5°F (38°C) or higher, a dry cough, and shortness of breath.

[0153] The illness reported from COVID-19 (i.e., caused by SARS-CoV-2) ranges from mild symptoms to severe illness and death in confirmed cases. The most common symptoms are fever, fatigue, dry cough, anosmia (loss of taste and / or smell), and shortness of breath. Runny nose, vomiting, diarrhea, skin rash (especially on the toes and fingers), sore throat, fatigue, muscle or body aches, headache, and sore throat have also been reported. These symptoms may appear 2 to 14 days after exposure.

[0154] Most people confirmed to have MERS-CoV infection had severe respiratory illness with symptoms of fever, cough, and / or shortness of breath. Some also had diarrhea and nausea / vomiting. Many people with MERS continued to experience more serious complications, such as pneumonia and kidney failure. Some infected individuals had mild symptoms (such as cold-like symptoms) or were asymptomatic.

[0155] In some embodiments, the subject has an underlying condition such as asthma, heart disease, diabetes, cancer, chronic lung disease, chronic heart disease, chronic kidney disease, or a combination thereof.

[0156] The age range of the subjects may be from young children, including pediatric subjects, to the elderly. In some embodiments, the subjects are at least 2 years of age, particularly if they have one or more symptoms, e.g., cough fever and / or other symptoms discussed herein. Treatment may be particularly indicated if the subject is male and / or over 40, 50, 60, 70 or 80 years of age. In some embodiments, the subjects are obese (BMI greater than 30, where BMI is the body mass index calculated as a person's weight (kilograms) divided by the square of their height (meters)).

[0157] In some embodiments, the subject has at least one mild or moderate COVID-19 symptom prior to treatment, such as fever or chills, cough, shortness of breath or difficulty breathing, fatigue, muscle or body aches, headache, novel loss of taste or smell, sore throat, congestion or runny nose, nausea, vomiting or diarrhea, for 6, 5, 4, 3, 2, or 1 day or less.

[0158] In other embodiments, the subjects are non-human mammals or birds. The symptoms caused by coronavirus infection in non-human species vary, causing upper respiratory tract illness in chickens, but diarrhea in cattle and pigs.

[0159] Coronavirus species and their representative viruses include: SARSr-CoV BtKY72 (Severe Acute Respiratory Syndrome-associated coronavirus), SARS-CoV-2 (Severe Acute Respiratory Syndrome-associated coronavirus), SARSr-CoV RaTG13 (Severe Acute Respiratory Syndrome-associated coronavirus), SARS-CoV PC4-227 (Severe Acute Respiratory Syndrome-associated coronavirus), SARS-CoV (Severe Acute Respiratory Syndrome-associated coronavirus), Bat-Hp-BetaCovC (Bat Hp-BetaCoronavirus Zhejiang 2013), Ro-BatCoV GCCDC1 (Rousettus bat betavirus GCCDC1), Ro-BatCoV HKU9 (Rousettus bat betavirus HKU9), Ei-BatCoV C704 (Eidolon bat coronavirus C704), Pi-BatCoV Examples include HKU5 (Pipistrellus bat coronavirus HKU5), Ty-BatCoV HKU4 (Tylonycteris bar coronavirus HKU4), MERS-CoV (Middle East Respiratory Syndrome-related coronavirus), EriCoV (Hedgehog coronavirus), MHV (mouse coronavirus), HCoV HKU1 (human coronavirus HKU1), ChRCoV HKU24 (China Rattus coronavirus HKU24), ChRCovC HKU24 (beta coronavirus 1), MrufCoV 2JL14 (Myodes coronavirus 2JL14), HCoV NL63 (human coronavirus NL63), HCoV 229E (human coronavirus 229E), and HCoV OC43 (human coronavirus OC43). For example, see Coronaviridae Study Group of the International Committee on Taxonomy of Viruses, Nat Microbiol 2020.DOI:10.1038 / s41564-020-0695-z), which incorporates the entire text specifically by reference. In some embodiments, coronaviruses are common cold coronaviruses such as 229E, NL63, OC43, and HKU1.

[0160] In a particularly preferred embodiment, the virus is a severe acute respiratory syndrome-associated virus, such as SARSr-CoV BtKY72, SARS-CoV-2, SARSr-CoV RaTG13, SARS-CoV PC4-227, or SARS-CoV, preferably a virus that infects humans, such as SARS-CoV or SARS-CoV-2.

[0161] In some embodiments, the virus is a Middle East Respiratory Syndrome-related virus such as MERS-CoV.

[0162] In some embodiments, the virus is SARS-CoV-2. The sequence WIV04 / 2019, belonging to the GISAID S clade / PANGO A lineage / Nextstrain 19B clade, is considered to most closely reflect the sequence of the original SARS-CoV-2 infecting humans. This is known as "sequence zero" and is widely used as a reference sequence. Following the initial isolation from Wuhan, China, numerous SARS-CoV-2 viral sequence variants of WIV 04 / 2019 have been identified, some of which may be particularly important due to their potential for increased infectivity, increased pathogenicity, and reduced vaccine effectiveness against them. SARS-CoV-2 variants with variations of the WIV04 / 2019 sequence include, but are not limited to, the following:

[0163] B.1.1.7 lineage (also known as 20I / 501Y.V1 Variant of Concern (VOC) 202012 / 01). This variant has a mutation in the receptor-binding domain (RBD) of the spike protein at position 501, where the amino acid asparagine (N) is replaced with tyrosine (Y). The abbreviation for this mutation is N501Y. This variant also has several other mutations, including the following: 69 / 70 deletion: Occurs spontaneously multiple times and likely leads to conformational changes in the spike protein. P681H: Near the S1 / S2 cleavage site, a region of high variability in coronaviruses. This mutation also appeared spontaneously multiple times.

[0164] Line B.1.351 (also known as 20H / 501Y.V2). This variant has multiple mutations in the spike protein, including K417N, E484K, and N501Y. Unlike line B.1.1.7 detected in the UK, this variant does not contain the 69 / 70 deletion.

[0165] The P.1 lineage (also known as 20J / 501Y.V3). The P.1 variant branched off from the B.1.1.28 lineage and was first reported by Japan's National Institute of Infectious Diseases (NIID) from four travelers from Brazil collected during routine screening at Tokyo's Haneda Airport. The P.1 lineage contains three mutations in the spike protein receptor-binding domain: K417T, E484K, and N501Y.

[0166] Other strains and variants of interest include, but are not limited to, B.1.1.207, B.1.429, B.1.427, B.1.525, and variants with two other mutations in the spike protein (e.g., N501T-G142D) or three mutations (e.g., N501T-G142D-F486 L). All of these strains and sequence substitutes compared to the WIV04 / 2019 strain are also considered SARS-CoV-2 viruses. In some embodiments, SARS-CoV-2 is a strain or isolate that poses a higher or potentially higher risk of causing human disease compared to WIV04 / 2019.For example, Science Brief, Emerging SARS-CoV-2 variants(CDC website,Updated Jan.28,2021), Horby et al., “NERVTAG note on B.1.1.7 severity” SAGE meeting report.January 21,2021; Wu et al., “mRNA-1273 vaccine induces neutralizing antibodies against spike mutants from global SARS-CoV-2 variants” bioRxiv.Posted January 25,2021; 4,2021];Weisblum et al., “Escape from See "neutralizing antibodies by SARS-CoV-2 spike protein variants" eLife 2020;9:e61312; and Resende et al., "Spike E484K mutation in the first SARS-CoV-2 reinfection case confirmed in Brazil" 2020. [Posted on virological.org on January 10, 2021].

[0167] Various strains and isolates of the aforementioned virus are publicly known, including, for example, representative genome sequences provided as Sequence IDs 1-4, accession numbers provided herein, and sequences and accession numbers provided by NCBI and GISAID, which provide hundreds of SARS-CoV-2 sequences, as well as, for example, Coronaviridae Study Group of the International Committee on Taxonomy of Viruses, Nat Microbiol 2020.DOI:10.1038 / s41564-020-0695-z).

[0168] In some embodiments, SARS-CoV-2 has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more mutations in its spike protein compared to another reference sequence, such as WIV04 / 2019, SEQ ID NO: 1 and / or SEQ ID NO: 2, or one provided therein. The spike protein sequence of SEQ ID NO: 1 is: [ka] [ka] This is coded in GenBank at 21563...25384:MN908947.3(sequence number 1), / gene=「S」, / note=「structural protein」, / codon_start=1 / product=「surface glycoprotein」, / protein_id=「QHD43416.1」).

[0169] In some embodiments, the SARS-CoV-2 spike protein has at least 70%, 75%, or 80%, preferably at least 85%, more preferably at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the spike protein of SEQ ID NO 1 or 2, or one or more spike proteins of other viral accessions provided herein.

[0170] In some embodiments, the SARS-CoV-2 spike protein has at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 5.

[0171] Mutations can be substitutions, insertions, deletions, or combinations thereof. Exemplary mutations discussed herein include, for example, one or more of the following mutations in the spike protein sequence: H69, and its absence in some cases; V70, and its absence in some cases; G142, and in some cases G142D; K417, and in some cases K417N or K417T; E484, and sometimes E484K; F486, and possibly F486L; and / or The N501 mutation, sometimes N501Y or N501T. These mutations are provided individually and in all combinations. These residues are shown in bold / italics / shading in Sequence ID No. 5 above.

[0172] In some embodiments, SARS-CoV-2 is of the B.1.1.7, B.1.351, P.1, B.1.1.207, B.1.429, B.1.427, or B.1.525 lineage.

[0173] In exemplary embodiments, SARS-CoV-2 is isolate USA / CA_CDC_5574 / 2020, or another isolate that shares one or more mutations with the original Wuhan isolate. Under the nomenclature system introduced by GISAID (Global Initiative on Sharing All Influenza Data), SARS-CoV-2, isolate USA / CA_CDC_5574 / 2020, was identified using the Phylogenetic Assignment of Named Global Outbreak LINeages (PANGOLIN) tool (GISAID website, 3. Rambaut et al., Nat. Microbiol. 5 (2020): 1403-1407. PubMed: 32669681; Mercatelli et al., Front. Microbiol. (2020): doi.org / 10.3389 / fmicb.2020.01800. PubMed: 32793182), lineage B.1.1.7 and GISAID clade The GR (Growth Rating) designation is assigned. The complete genome of SARS-CoV-2 and isolate USA / CA_CDC_5574 / 2020 have been sequenced (GISAID:EPI_ISL_751801). The following mutations exist in the clinical isolate:

[0174] Spike A570D, Spike D614G, Spike D1118H, Spike H69del, Spike N501Y, Spike P681H, Spike S982A, Spike T716I, Spike V70del, Spike Y145del, M (membrane protein) V70L, N (nucleocapsid protein) D3L, N G204R, N R203K, N S235F, NS3 T223I, NS8 (unstructured protein 8) Q27stop, NS8 R52I, NS8 Y73C, NSP3 (unstructured protein 3) A890D, NSP3 I1412T, NSP3 T183I, NSP6 (unstructured protein 6) F108del, NSP6 G107del, NSP6 S106del, NSP12(unstructured protein 12)P323L, NSP13(unstructured protein 13)A454V, NSP13 K460R. One further SNP in ORF1ab L3826F was reported in deposited passage 2 viruses compared to clinical specimens. See also BEI Resources, catalog number NR-54011, and its description, which are incorporated herein by reference in their entirety.

[0175] However, these are non-limiting examples, and the disclosed compositions and methods can also be used to treat other strains of coronavirus, particularly SARS and MERS coronaviruses. In some embodiments, the viral genome (DNA sequence) has at least 70%, 75%, or 80%, preferably at least 85%, more preferably at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NOs. 1, 2, 3, or 4 provided herein, or one or more other viral accessions, or the sequence or accession provided in Coronaviridae Study Group of the International Committee on Taxonomy of Viruses, Nat Microbiol 2020.DOI:10.1038 / s41564-020-0695-z (all of which are incorporated herein by reference in their entirety). While the sequences are provided as DNA sequences, it will be understood that the viral genome itself typically has a corresponding RNA sequence. Therefore, the corresponding RNA sequences are also explicitly provided herein.

[0176] GenBank accession number MN908947.3 and NCBI accession number NC_045512.2, which are incorporated herein by reference in their entirety, provide the (DNA) genome sequence of SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2 isolate Wuhan-Hu-1, complete genome): (Sequence ID 1).

[0177] GenBank accession number MN985325.1, which is incorporated herein by reference in its entirety, provides the (DNA) genome sequence for SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2 isolate 2019-nCoV / USA-WA1 / 2020, complete genome); (Sequence ID 2).

[0178] GenBank accession number GenBank:AY274119.3 is incorporated herein by reference in its entirety and provides the (DNA) genome sequence of SARS-CoV (Severe Acute Respiratory Syndrome-related Coronavirus isolate Tor2, complete genome) (SEQ ID NO: 3).

[0179] GenBank accession number GenBank:JX869059.2 is incorporated herein by reference in its entirety and provides the (DNA) genome sequence for MERS-CoV (human beta-coronavirus 2c EMC / 2012, complete genome) (SEQ ID NO: 4).

[0180] In some embodiments, a subject is diagnosed with a positive SARS-CoV-2 virus test result and has had at least one mild or moderate COVID-19 symptom (i.e., fever or chills, cough, shortness of breath or difficulty breathing, fatigue, muscle or body aches, headache, novel loss of taste or smell, sore throat, congestion or runny nose, nausea, vomiting or diarrhea) for six days or less prior to the first administration of probenecid, its metabolite or analogue, or a pharmaceutically acceptable salt thereof.

[0181] The disclosed compositions and methods can be further understood through the following numbered sections.

[0182] 1. A method for treating a subject with coronavirus infection, comprising administering an effective amount of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof, to the subject.

[0183] 2. The method according to item 1, wherein the method includes administering probenecid to the target.

[0184] 3. The method according to claim 1 or 2, wherein probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof is present in a pharmaceutical composition further comprising a pharmaceutically acceptable carrier and / or excipient.

[0185] 4. The method according to any one of items 1 to 3, comprising systemic administration of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof.

[0186] 5. The method according to any one of items 1 to 4, for administering probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof orally, parenterally, topically, or mucosally.

[0187] 6. The method according to any one of items 1 to 4, comprising orally administering probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof.

[0188] 7. The method according to any one of items 1 to 4, wherein probenecid, its metabolite or analogue, or a pharmaceutically acceptable salt thereof is administered into the lungs (e.g., pulmonary administration).

[0189] 8. The method according to any one of items 1 to 4, comprising intranasal administration of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof.

[0190] 9. The method according to any one of items 1 to 8, comprising administering probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof, in an amount effective in reducing viral replication.

[0191] 10. The method according to any one of items 1 to 9, comprising administering probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof, in an amount effective in reducing one or more symptoms of a virus-related illness, disorder, or disease.

[0192] 11. The method according to any one of items 1 to 10, wherein the symptoms include fever, sinus and / or pulmonary congestion, runny nose or nasal congestion, cough, sneezing, sore throat, body aches, fatigue, shortness of breath, chest tightness, wheezing on exhalation, chills, muscle pain, headache, diarrhea, malaise, nausea, vomiting, anosmia, skin rash, and combinations thereof.

[0193] 12. The method according to any one of items 1 to 11, wherein the virus is severe acute respiratory syndrome-associated coronavirus, bat Hp-betacoronavirus Zhejiang2013, Rousettus bat betavirus GCCDC1, Rousettus bat coronavirus HKU9, Eidolon bat coronavirus C704, Pipistrellus bat coronavirus HKU5, Tylonycteris bar coronavirus HKU4, Middle East respiratory syndrome-associated coronavirus, Hedgehog coronavirus, mouse coronavirus, human coronavirus HKU1, China Rattus coronavirus HKU24, betacoronavirus 1, Myodes coronavirus 2JL14, human coronavirus NL63, human coronavirus 229E, or human coronavirus OC43.

[0194] 13. The method described in paragraph 12, wherein the virus is a severe acute respiratory syndrome-associated coronavirus.

[0195] 14. The method according to paragraph 13, wherein the severe acute respiratory syndrome-associated coronavirus is SARS-CoV-2, SARS-CoV, SARSr-CoV RaTG13, SARS-CoV PC4-227, or SARSr-CoV BtKY72.

[0196] 15. The method described in paragraph 14, wherein the coronavirus associated with severe acute respiratory syndrome is SARS-CoV-2.

[0197] 16. The method according to item 15, wherein SARS-CoV-2 comprises a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1 or 2.

[0198] 17. The method described in paragraph 14, wherein the coronavirus associated with severe acute respiratory syndrome is SARS-CoV.

[0199] 18. The method according to item 17, wherein the SAR-CoV comprises a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 3.

[0200] 19. The method described in paragraph 12, wherein the virus is Middle East Respiratory Syndrome-related coronavirus.

[0201] 20. The method described in paragraph 19, wherein the Middle East Respiratory Syndrome-related coronavirus is MERS-CoV.

[0202] 21. The method according to item 20, wherein MERS-CoV comprises a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 4.

[0203] 22. The method described in any one of items 1 through 21, wherein the subject is exposed to the coronavirus.

[0204] 23. The method described in item 22, wherein the subject is currently infected with the coronavirus.

[0205] 24. The method described in paragraph 23, wherein the subject has COVID-19.

[0206] 25. The method described in any one of items 1 to 20, wherein the subject is asymptomatic.

[0207] 26. The method described in any one of items 1 through 25, wherein the subject is exposed to the coronavirus.

[0208] 27. The method described in any one of paragraphs 1 to 26, wherein the treatment is for the prevention of coronavirus infection in the subject.

[0209] 28. The method described in item 27, wherein the subject had close contact with a person infected with the coronavirus.

[0210] 29. A method for treating a subject infected with SAR-CoV-2, comprising administering an effective amount of probenecid or a pharmaceutically acceptable salt thereof to the subject.

[0211] 30. The method described in paragraph 29, wherein the subject has COVID-19.

[0212] 31. A preventive method comprising administering an effective dose of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof, to subjects not exposed to the coronavirus, thereby reducing viral infection in subjects upon exposure to the virus compared to viral infection in the absence of treatment.

[0213] 32. A method for preventing coronavirus infection, comprising administering an effective amount of probenecid or a pharmaceutically acceptable salt thereof to a subject to reduce coronavirus infection in the subject before exposure to the coronavirus.

[0214] 33. The method according to any one of paragraphs 29 to 32, wherein the coronavirus is SAR-CoV-2.

[0215] 34. The method according to any one of items 1 to 33, wherein the subject is administered 10 mg to 1,000 mg or 50 mg to 500 mg of probenecid, its metabolites or analogs, or a pharmaceutically acceptable salt thereof, once to five times daily until the symptoms are relieved, the infection is resolved, or a combination thereof.

[0216] 35. The method according to any one of items 1 to 34, wherein the subject is treated by pulsed administration.

[0217] 36. The method according to item 35, wherein the pulse administration comprises one to five bolus doses of probenecid, its metabolites or analogs, or pharmaceutically acceptable salts thereof, ranging from 1,000 mg to 5,000 mg.

[0218] 37. The method according to item 36, wherein, after a bolus administration, a drug-free period is followed, if necessary, until serum levels of probenecid, its metabolites or analogues, or pharmaceutically acceptable salts thereof are approximately zero.

[0219] 38. The method according to any one of the items 35 to 37, wherein probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof, is administered orally or by infusion.

[0220] 39. A pharmaceutical composition comprising an effective amount of probenecid, its metabolites or analogues, or a pharmaceutically acceptable salt thereof, for use in the manner described in any one of items 1 to 39.

[0221] 40. A compound for use in the treatment of coronavirus infection in a person requiring treatment for coronavirus infection, wherein the compound is probenecid or its metabolite or analogue or a pharmaceutically acceptable salt thereof.

[0222] 41. A compound for use as described in item 40, wherein the subject has one or more symptoms of a disease, disorder or illness related to coronavirus, and the symptoms are selected as such from fever, sinus and / or pulmonary congestion, runny nose or nasal congestion, cough, sneezing, sore throat, body aches, fatigue, shortness of breath, chest tightness, wheezing on exhalation, chills, muscle aches, headache, diarrhea, malaise, nausea, anosmia, skin rash and combinations thereof.

[0223] 42. Compounds for use as described in subsection 40 or 42, wherein the subject has COVID-19.

[0224] 43. Compounds for use as described in item 40, wherein the subject is asymptomatic.

[0225] 44. Compounds for use as described in item 40, for which the treatment is for the prevention of coronavirus infection in the subject.

[0226] 45. Compounds for use as described in item 44, in which the subject has had close contact with a person infected with the coronavirus.

[0227] 46. ​​A compound for use according to any one of claims 40 to 45, wherein the compound is present in a pharmaceutical composition further comprising a pharmaceutically acceptable carrier and / or excipient.

[0228] 47. A compound for use as described in any one of sub-sub

[0229] 48. A compound for use as described in any one of sub-sub

[0230] 49. The compound for use as described in item 48, wherein the treatment involves mucosal administration of the compound to the lungs, nasal mucosa, or a combination thereof.

[0231] 50. A compound for use as described in any one of sub-sub

[0232] 51. A compound for use as described in any one of items 40 to 50, wherein the coronavirus is Severe Acute Respiratory Syndrome-related Coronavirus, Bat Hp-Beta Coronavirus Zhejiang2013, Rousettus Bat Betavirus GCCDC1, Rousettus Bat Coronavirus HKU9, Eidolon Bat Coronavirus C704, Pipistrellus Bat Coronavirus HKU5, Tylonycteris Bar Coronavirus HKU4, Middle East Respiratory Syndrome-related Coronavirus, Hedgehog Coronavirus, Mouse Coronavirus, Human Coronavirus HKU1, China Rattus Coronavirus HKU24, Beta Coronavirus 1, Myodes Coronavirus 2JL14, Human Coronavirus NL63, Human Coronavirus 229E, or Human Coronavirus OC43.

[0233] 52. A compound for use as described in any one of subsections 40 to 51, wherein the coronavirus is a severe acute respiratory syndrome-associated coronavirus, and in some cases the severe acute respiratory syndrome-associated coronavirus is SARS-CoV-2, SARS-CoV, SARSr-CoV RaTG13, SARS-CoV PC4-227, or SARSr-CoV BtKY72.

[0234] 53. The compounds for use described in Section 52, wherein the severe acute respiratory syndrome-associated coronavirus is SARS-CoV-2, and optionally, SARS-CoV-2 comprises a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1 or 2.

[0235] 54. The treatment is, (i) administer to the subject 10 mg to 1,000 mg or 50 mg to 500 mg of probenecid or a pharmaceutically acceptable salt thereof, twice daily as needed, for 14 days as needed; and (ii) A pulse dosing regimen, wherein the pulse dosing, if necessary, includes one to five bolus doses of 1,000 mg to 5,000 mg of probenecid or a pharmaceutically acceptable salt thereof, followed by a rest day if necessary. A compound for use as described in any one of the items in items 40 to 53, comprising one or more of the following:

[0236] 55. A compound or method according to any of the preceding items, wherein SARS-CoV-2 comprises a spike protein having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 5.

[0237] 56. A compound or method according to any of the preceding items, wherein SAR-CoV-2 comprises one or more mutations in SEQ ID NO: 5, selected from H69, optionally its deletion; V70, optionally its deletion; G142, optionally G142D; K417, optionally K417N or K417T; E484, optionally E484K; F486, optionally F486L; and the N501 mutation, optionally N501Y or N501T.

[0238] 57. A compound or method described in any of the preceding sections, wherein SARS-CoV-2 is derived from lineage B.1.1.7, B.1.351, P.1, B.1.1.207, B.1.429, B.1.427, or B.1.525. [Examples]

[0239] Example 1: Probenecid reduces SARS-CoV-2 plaque formation in vitro. material and method Plaque reduction assay Preventive measures Vero E6 cells (ATCC CRL-1586) were divided into 5 × 10⁻¹⁶ cells. 5 The cells were plated in 12-well plates at the specified cell / well ratio and incubated overnight.

[0240] Cells were washed once with PBS, and probenecid was added to the wells in the culture medium at concentrations of 0.1 μM, 1 μM, 2.5 μM, or 5 μM, or a vehicle control was added, and incubated for 24 hours (Figure 1). All wells were normalized to 0.05% DMSO. Each concentration was completed in a double-row system.

[0241] After pretreatment, the culture medium was discarded, and the cells were refilled with medium containing probenecid (as described above) and SARS-CoV-2 (a stock grown from isolate USA-WA1 / 2020, BEI Resources Catalogue Ref. Number NR-52281). The complete genome of SARS-CoV-2, USA-WA1 / 2020, was sequenced (isolate - GenBank accession number: MN985325 and after 1 passage in Vero cells - GenBank accession number: MT020880 and after 4 passages in Vero cells - GenBank accession number: MT246667).

[0242] Cells were infected with a MOI of 0.01 for 4 days. After infection, the cells were fixed and stained to visualize the plaques. The plaques were manually quantified (Figure 1).

[0243] Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparison test (treatment vs. DMSO). A repeated measures design was used, with P<0.05. * .

[0244] In another experiment, probenecid at 0.0001 μM, 0.001 μM, 0.01 μM, 0.1 μM, 1 μM, 2.5 μM or 5 μM (Figure 2) was added. Vero E6 cells were plated at 8E5 cells / well in a 6-well plate and incubated overnight. The cells were washed once with PBS, the compound was added to the wells in culture medium, and incubated for 24 hours. All wells were normalized to 0.05% DMSO. Each concentration was done in duplicate. After pretreatment, the medium was discarded and the cells were replenished with medium containing the drug (as described above) and SARS-CoV-2. The cells were infected with the virus at an MOI of 0.01 for 4 days. After infection, the cells were fixed and stained to visualize the plaques. The plaques were quantified (Figure 2).

[0245] Therapeutic treatment Vero E6 cells were plated at 8E5 cells / well in a 6-well plate and incubated overnight. The cells were washed once with PBS and infected with the virus at an MOI of 0.01 for 1 hour. After 1 hour of infection, probenecid at 0.0001 μM, 0.001 μM, 0.01 μM, 0.1 μM, 1 μM, 2.5 μM or 5 μM was added to the wells in overlay medium and incubated for 4 days. After infection, the cells were fixed and stained to visualize the plaques. The plaques were quantified (Figure 3).

[0246] Results The effect of probenecid on virus replication was examined using an in vitro plaque formation assay.

[0247] Prolonged pretreatment resulted in a dose-dependent decrease in plaque formation at the tested concentrations (5 μM to 0.1 μM) in two independent experiments. 5 μM to 0.1 μM reduced plaque formation by approximately 89% to 72% respectively compared to the DMSO-treated infection control (Figure 1).

[0248] In another experiment, probenecid pretreatment resulted in a dose-dependent decrease in plaque formation at the tested concentrations (5 μM to 0.0001 μM) in two independent experiments. 5 μM to 0.0001 μM decreased plaque formation by approximately 93% to 50% respectively compared to the DMSO-treated infection control (Figure 2).

[0249] In another experiment, probenecid post-treatment resulted in a dose-dependent decrease in plaque formation at the tested concentrations (5 μM to 0.0001 μM). 5 μM to 0.0001 μM decreased plaques by approximately 90% to 40% respectively compared to the DMSO-treated infection control (Figure 3).

[0250] These results indicate that probenecid significantly reduces virus titer / plaque formation with a 24-hour pretreatment at the tested concentrations. These results also indicate that probenecid significantly reduces virus titer / plaque formation after treatment at the tested concentrations.

[0251] Example 2: Preventive and therapeutic probenecid treatment reduces viral load in vivo Materials and methods Eighty-four hamsters were obtained and used as subjects for an assay to determine whether probenecid has a preventive effect and / or a therapeutic effect against SARS-CoV-2 (stock grown from isolate USA-WA1 / 2020, BEI Resources Catalogue Ref. Number NR-52281) infection in vivo.

[0252] Preventive probenecid treatment was performed on day -1. Infection (1×10 3 PFU) was induced by delivering the virus intranasally in a 50 μl volume on day 0. Post-infection / therapeutic probenecid treatment was on day 2. The animals were euthanized and tissues were collected for analysis on days 0, 3, and 7. See Figure 5.

[0253] Group · Non-infected (n = 2 / time point) • PBS infection (n=2 / time period) • Prophylactic, administered 24 hours prior to infection, 200 mg / kg (n=6 / time point in time) • Prophylactic, administered 24 hours prior to infection, 2 mg / kg (n=6 / time point in time) Treatment, 48 hours after infection, • 200 mg / kg (n=6 / at this point in time) Treatment: 48 hours after infection, 2 mg / kg (n=6 / time point in time) n=3 time points (days 0, 3, and 7 after infection) n = a total of 84 animals

[0254] endpoint • Weight and clinical symptoms (daily) • Viral titer / load (n=3 / group / time point) • Homogenize the lungs • Titer (Plaque assay and TCID) 50 ) • RNA extraction and qPCR • Histopathology (n=3 / group / time point) • Phlebotomy (to store serum for neutralization assays, ELISA, and qPCR) • Expand with formalin solution • Fixed in formalin for more than 72 hours • H&E staining, reading

[0255] TCID 50 Assay TCID 50 The procedure was completed in a 96-well plate of Vero E6 cells. Lung homogenate was serially diluted and 200 μl was added to each well. The plate was incubated for 72 hours. The plate was then fixed and stained.

[0256] Plaque assay Plaque assays were completed in 12-well plates of Vero E6 cells. Serially diluted lung homogenate was added to each well at a rate of 500 μl. The plates were incubated for 96 hours. The plates were then fixed and stained.

[0257] result The hamsters' weight was measured daily before treatment and then after infection (day 0). The uninfected group continuously gained weight throughout the study. Other animals stopped gaining weight after infection but did not experience significant weight loss. See Figure 5.

[0258] The results indicate that no virus was detected in the lungs of uninfected hamsters. On post-infection (pi) day 3, all probenecid-treated hamsters had lower lung virus titers (1E5) compared to PBS-treated hamsters (1E9+). The results indicate that probenecid reduces lung virus titers by several logarithms. Treatments of 2 mg and 200 mg were equally effective, even though the hamsters received a single dose. Prophylactic and therapeutic treatments worked similarly. TCID 50 The plaque data were nearly identical. No lung virus was detectable on day 7 post-infection. See Figures 6 and 7.

[0259] Example 3: Probenecid improves recovery from SARS-CoV-2 infection in human subjects. material and method In an open-label, physician-initiated clinical trial (IIS) involving 10 non-hospitalized patients with mild to moderate SARS-CoV-2 infection, treated with probenecid for 28 days ("Clinical Trial").

[0260] Selection criteria: Non-hospitalized patients diagnosed with a positive SARS-CoV-2 RNA test within 5 days prior to screening / baseline / first dose visit 1. • The patient developed at least one mild or moderate COVID-19 symptom (i.e., fever or chills, cough, shortness of breath or difficulty breathing, fatigue, muscle or body aches, headache, novel loss of taste or smell, sore throat, congestion or runny nose, nausea, vomiting or diarrhea) within 5 days or less of screening. • Men and women aged 18 and over. · The patient has an estimated glomerular filtration rate (eGFR) of 30 mL / min or more, estimated using the Cockcroft-Gault formula. · The patient is willing to participate in this study, sign an informed consent, and participate in regular follow-up during the study. · The patient can understand the study procedures and cooperate.

[0261] Exclusion criteria: · The patient is hospitalized or requires hospitalization at the time of registration. · The patient is in an immunocompromised state due to human immunodeficiency virus (HIV) or other "immunocompromised" conditions. · The patient has severe cognitive impairment or mental illness. · Women who are capable of giving birth, are breastfeeding, plan to become pregnant or breastfeed during the study period, or have a positive urine pregnancy test at screening (negative results are required for eligibility). · Patients who may be allergic to the investigational drug and for whom the PI determines that the investigational drug is not appropriate for the patient. · The patient is simultaneously participating in another clinical trial. · Patients with a medical condition that may interfere with drug metabolism or absorption in the opinion of the principal investigator of the clinical trial (e.g., short bowel syndrome, Crohn's disease, etc.). · There is evidence of a major medical condition or laboratory finding that, in the opinion of the principal investigator of the clinical trial, makes it undesirable for the patient to participate in the trial, such as a history of malignancy, uric acid kidney stones, known blood disorders, or other unstable / clinically important diseases. · The patient has a known history of chronic obstructive pulmonary disease (COPD). · The patient has received any investigational treatment with anti-COVID-19 drugs, such as remdesivir, camostat, ritonavir, hydroxychloroquine, azithromycin, ruxolitinib, or corticosteroids, in the past 30 days. · The patient has received an organ transplant, stem cell transplant, or bone marrow transplant.

[0262] Analysis variables: Screening variables consist of baseline patient characteristics: complete physical examination, height and weight, vital signs, hematology, chemistry panel, and urine pregnancy test. The effectiveness variables consist of qualitative SARS-CoV-2 testing, SpO2, body temperature and respiratory symptom grading (WHO clinical status on an ordinal scale), time to hospitalization, and length of hospital stay. Routine and special safety variables include AE / SAE assessment, clinical laboratory assessments: CBC, CMP, TG, AA, OM-3 ratio, EPA and DHA, hs-CRP, LDH, CPK, ALT, creatinine, ferritin, fibrinogen, D-dimer, and von Willebrand factor (vWF) antigen.

[0263] COVID-19 positive patients were screened, physically examined on day 1, and retested using iAMP qualitative SARS-CoV-2 RNA testing. Safety clinical laboratory tests were also performed on day 1, including CBC by differential and comprehensive metabolic profile (CMB) and inflammatory markers for COVID-19 (i.e., high-sensitivity (hr-CRP), ferritin, d-dimer, CPK, LDH).

[0264] On day 2, after reviewing safety clinical laboratory values, the principal investigator determined whether the patient could continue the study and administered the initial dose of the investigational drug probenecid 500 mg. The patient received 500 mg twice daily for 14 days, from day 2 to day 15 of the study.

[0265] The patients returned to the hospital on days 5, 10, and 15 of the study, and completed the study on day 28.

[0266] The age demographics in this study ranged from 42 to 81 years. Patients 1, 3, and 8 had pre-existing comorbidities. Patients enrolled in the study had the characteristics of Table 1 and Figures 8A and 8B.

[0267] [Table 1]

[0268] result A clinical trial was designed to test the efficacy of probenecid against SARS-CoV-2 infection in human subjects. Data including positive / negative SARS-CoV-2 testing, symptom improvement, safety, coagulation, inflammatory markers, and adverse events were monitored in 10 patients treated over a 14-day period (day 15), and these are discussed in more detail below.

[0269] SARS-CoV-2 testing In a recent in silico study, the authors conducted a literature search to understand SARS-CoV-2 viral shedding. 77 studies / reports were eligible to be included. This review found that the pooled median duration of viral shedding from respiratory sources for mild to moderate illness was 17.2 days (Cevik et al., "SARS-CoV-2, SARS-CoV, and MERS-CoV viral load dynamics, duration of viral shedding, and infectiousness: a systematic review and meta-analysis," The Lancet, 2(1), E13-E22 (2021), DOI:10.1016 / S2666-5247(20)30172-5). In patients treated with probenecid from this IIS study, viral shedding was lower than the pooled median for all patients except one patient, patient 003.

[0270] Of the 10 patients enrolled in the study who tested positive for SARS-CoV-2, 2 tested negative on day 4 of treatment, 4 on day 9, and 9 on day 14.

[0271] The results of the SARS-CoV-2 negative test are summarized in Figure 9. Observation of weight as a comorbidity showed that patient 003 weighed over 300 pounds and remained positive on day 14 of treatment.

[0272] Symptom improvement The improvement in symptoms is summarized in Figure 10. Two symptoms persisted: shortness of breath and loss of taste or smell. The persistence of these symptoms over time is not uncommon in SARS-CoV-2 infection.

[0273] Patient 001, an 81-year-old male with comorbid hypertension, was hospitalized for shortness of breath from day 5 to day 11 of the procedure. The patient continued taking the investigational drug, received 2-3 liters of oxygen during his stay, and received 2 liters of oxygen via nasal cannula after returning home. During his hospitalization, the patient was administered dexamethasone and azithromax, but not convalescent plasma or remdesivir.

[0274] Indicator outcomes and inflammatory responses Low lymphocyte counts, elevated fibrinogen and D-dimer levels may be indicators of a worse outcome. Patient 001 had low lymphocyte counts from enrollment until day 15 of the study, at which point they returned to normal. Patient 001 also had elevated fibrinogen and D-dimer levels until day 15 of the study, and elevated LDH on day 15.

[0275] Patient 005 also entered the emergency room on day 15 of the study. This woman was diagnosed with pneumonia, but was discharged home because her oxygen saturation was normal. She was administered steroids and antibiotics. Patient 005 had elevated D-dimer levels up to day 15 of the study.

[0276] Figure 11 shows the temperature analysis (resolution) across all patients.

[0277] High-sensitivity C-reactive protein (hs-CRP) is an indicator of the initial inflammatory response to SARS-CoV-2. Values ​​>3.1 indicate infection and inflammation. hs-CRP levels surged in two patients but decreased during treatment. By day 15 of the study (day 14 of treatment), hs-CRP levels had not returned to normal. Figure 12 shows hs-CRP levels and treatment response for all patients.

[0278] D-dimer is another indicator of early coagulation disorders and inflammatory responses associated with SARS-CoV-2 infection. The normal range for d-dimer is less than 0.5 mcg / ml. All 10 patients showed signs of SARS-CoV-2-related early coagulation disorders and inflammation. Eight of the 10 patients had levels less than 1 mcg / ml during the treatment phase of the study. Patient 003 had the highest value at enrollment and approached the normal range during treatment. Patient 001's d-dimer level remained high until day 15 of the study. Figure 13 shows the d-dimer levels of all patients during treatment.

[0279] Elevated fibrinogen levels exceeding 425 mg / dL are an indicator of coagulation or thrombosis. Three patients had fibrinogen levels above the upper limit of normal. As described above, patient 001 had elevated fibrinogen levels until day 15 of the study. Patient 007 had elevated fibrinogen levels, which decreased to normal by day 9 of treatment, and patient 008's levels decreased to normal by day 14 of treatment. Figure 14 shows the fibrinogen levels for all patients.

[0280] In this age group, elevated lactate dehydrogenase (LDH) levels exceeding 250 U / L are indicative of tissue damage. Patient 001, who required hospitalization and oxygen, had elevated LDH levels after discharge. Patient 010 had slightly elevated levels on day 14 of treatment. LDH levels for all patients are shown in Figure 15.

[0281] conclusion This open-label study of 10 consecutively enrolled non-hospitalized patients showed that probenecid treatment reduced SARS-CoV-2 viral shedding to below the pooled median of 17.2 days (Cevik et al., "SARS-CoV-2, SARS-CoV, and MERS-CoV viral load dynamics, duration of viral shedding, and infectiousness: a systematic review and meta-analysis," The Lancet, 2(1), E13-E22 (2021), DOI:10.1016 / S2666-5247(20)30172-5). Although a scored grading system was not used, patients' symptoms showed improvement. Coagulation and inflammatory markers remained mostly within normal ranges, with the exception of one patient (patient 001) who had elevated biomarkers at enrollment.

[0282] Example 4: Probenecid shows favorable comparative results with remdesivir in an in vitro plaque assay. Remdesivir is a nucleotide prodrug of adenosine analog antiviral drug approved by the FDA on October 22, 2020, for the treatment of COVID-19 requiring hospitalization. Experiments were designed to compare the antiviral activity of probenecid, remdesivir, and the combination of probenecid and remdesivir. Prophylactic and therapeutic (at the time of infection) experiments were generally performed in NHBE and Vero cells as described above in Example 1. All experiments were performed in triplicate in more than three independent cycles. The results are shown in Figures 16A to 20F.

[0283] The results showed a dose-dependent reduction in viral plaque formation when prophylactically or therapeutically treated with probeniside compared to the control group.

[0284] Prophylactic treatment with probenecid inhibits SARS-CoV-2 replication in mammalian cells (for example, Vero E6 cells treated with different concentrations of probenecid inhibited SARS-CoV-2 replication, similar to the results presented in Example 1).

[0285] Under prophylactic and therapeutic conditions, probenecid had lower IC50 and IC90 values ​​than remdesivir in both cell types tested.

[0286] The results also indicate that co-administration of 0.1 μM probenecid with remdesivir enhances the efficacy of remdesivir in CoV2-infected NHBE cells.

[0287] Example 5: Probenecid reduces SARS-CoV-2 B.1.1.7 plaque formation in vitro. material and method Genome RNA from SARS-related coronavirus 2, isolate USA / CA_CDC_5574 / 2020 (lineage B.1.1.7) (BEI Resources, Catalog No. NR-55244) SARS-CoV-2 isolate USA / CA_CDC_5574 / 2020 was isolated from a nasopharyngeal swab in San Diego County, California, USA on December 29, 2020 (see, for example, the GISAID website). Under the nomenclature system introduced by GISAID (Global Initiative on Sharing All Influenza Data), SARS-CoV-2, isolate USA / CA_CDC_5574 / 2020, was identified using the Phylogenetic Assignment of Named Global Outbreak LINeages (PANGOLIN) tool (GISAID website, 3. Rambaut et al., Nat. Microbiol. 5(2020):1403-1407. PubMed:32669681; Mercatelli et al., Front. Microbiol. (2020):doi.org / 10.3389 / fmicb.2020.01800. PubMed:32793182) as lineage B.1.1.7 and GISAID clade The GR (Growth Leader) designation is assigned. The complete genome of SARS-CoV-2, isolate USA / CA_CDC_5574 / 2020, has been sequenced (GISAID:EPI_ISL_751801).

[0288] The following mutations are present in clinical isolates: spike A570D, spike D614G, spike D1118H, spike H69del, spike N501Y, spike P681H, spike S982A, spike T716I, spike V70del, spike Y145del, M (membrane protein) V70L, N (nucleocapsid protein) D3L, N G204R, N R203K, N S235F, NS3 T223I, NS8 (unstructured protein 8) Q27stop, NS8 R52I, NS8 Y73C, NSP3 (unstructured protein 3) A890D, NSP3 I1412T, NSP3 T183I, NSP6 (unstructured protein 6) F108del, NSP6 G107del, NSP6 S106del, NSP12(non-structural protein 12)P323L, NSP13(non-structural protein 13)A454V, NSP13 K460R. One additional SNP in ORF1ab L3826F was reported in deposited passage 2 viruses compared to clinical specimens.

[0289] hCoV-19 / Wuhan / WIV04 / 2019(WIV04) is the official reference sequence used by GISAID(EPI_ISL_402124). The reason for selecting WIV04 was its high-quality genome sequence and its representation of one of the few early deposits of a betacoronavirus that causes COVID-19 (Okada et al., Euro Surveill., 25(8), 5 pages (2020) pii=2000097. DOI: 10.2807 / 1560-7917. ES. 2020.25.8.2000097). WIV04 is representative of the early outbreak sequences and is identical to it. WIV04 was isolated by the Wuhan Institute of Virology from a clinical sample of bronchoalveolar lavage fluid (BALF) collected on December 30, 2019, at Jinyintan Hospital in Wuhan, Hubei Province, from a symptomatic retailer working at the Huanan Seafood Wholesale Market.

[0290] Infection Time Assay Vero cells were tested in a 12-well format at the time of infection. Probenecid was evaluated at 100, 50, 25, 12, 6, 3, 1, 0.1, 0.01, 0.001, 0.0001, and 0.00001 μM. The assay was continued for 3 days. Cells were fixed and stained to visualize plaques. Plaques were quantified.

[0291] result The in vitro plaque assay was performed as described above, using SARS-CoV-2 isolate USA / CA_CDC_5574 / 2020 as the SARS-CoV-2 test virus. This variant of the original SARS-CoV-2 isolate is classified as a variant of concern due to increased infectivity.

[0292] The data in Figure 21 shows that probenecid has a similar reduction in viral titer in SARS-CoV-2 isolate hu / USA / CA_CDC_5574 / 2020 compared to SARS-CoV-2 strain 2019 nCoV / USA-WA1 / 2020, as shown in Example 1 and related figures.

[0293] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art in the field to which the inventions of this disclosure pertain. Publications cited herein and the materials they refer to are incorporated more specifically by reference.

[0294] Those skilled in the art will recognize many equivalents to the specific embodiments of the invention described herein, or will be able to confirm them simply by routine experimentation. Such equivalents are intended to be covered by the following claims. In certain embodiments, for example, the following are provided: (Item 1) A method for treating a subject infected with severe acute respiratory syndrome coronavirus (SARS-CoV-2), comprising administering to the subject requiring said treatment an effective amount of a compound selected from probenecid, its metabolites, its analogues, and any pharmaceutically acceptable salt thereof. (Item 2) The method according to item 1, wherein the subject has one or more symptoms of the diseases, disorders, or illnesses associated with SARS-CoV-2. (Item 3) The method according to item 2, wherein one or more of the symptoms are selected from fever, sinus and / or pulmonary congestion, runny nose or nasal congestion, cough, sneezing, sore throat, body aches, fatigue, shortness of breath, chest tightness, wheezing on exhalation, chills, muscle pain, headache, diarrhea, malaise, nausea, anosmia, skin rash, and combinations thereof. (Item 4) The method described in item 1, wherein the subject has COVID-19. (Item 5) The method according to item 1, wherein the subject is asymptomatic. (Item 6) The method according to item 1, wherein the SARS-CoV-2 comprises a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1 or 2. (Item 7) The method according to item 1, wherein the SARS-CoV-2 comprises a spike protein having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 5. (Item 8) The method according to item 1, wherein the SAR-CoV-2 comprises one or more spike protein mutations for SEQ ID NO: 5, selected from H69, optionally its deletion; V70, optionally its deletion; G142, optionally G142D; K417, optionally K417N or K417T; E484, optionally E484K; F486, optionally F486L; and the N501 mutation, optionally N501Y or N501T. (Item 9) The method according to item 1, wherein the SARS-CoV-2 is derived from lineage B.1.1.7, B.1.351, P.1, B.1.1.207, B.1.429, B.1.427, or B.1.525. (Item 10) The method according to item 1, wherein the compound is probenecid or a pharmaceutically acceptable salt thereof. (Item 11) A method for preventing infection with severe acute respiratory syndrome coronavirus (SARS-CoV-2), comprising administering to a subject requiring such prevention an effective amount of a compound selected from probenecid, its metabolites, its analogues, and any pharmaceutically acceptable salt thereof. (Item 12) The method according to item 11, wherein the subject is exposed to SARS-CoV-2. (Item 13) The method described in item 11, wherein the subject had close contact with a person infected with SARS-CoV-2. (Item 14) The method according to item 11, wherein the SARS-CoV-2 comprises a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1 or 2. (Item 15) The method according to item 11, wherein the SARS-CoV-2 comprises a spike protein having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 5. (Item 16) The method according to item 11, wherein the SAR-CoV-2 comprises one or more (sphmore) mutations in sequence number 5 selected from H69, optionally its deletion; V70, optionally its deletion; G142, optionally G142D; K417, optionally K417N or K417T; E484, optionally E484K; F486, optionally F486L; and the N501 mutation, optionally N501Y or N501T. (Item 17) The method according to item 11, wherein the SARS-CoV-2 is derived from lineage B.1.1.7, B.1.351, P.1, B.1.1.207, B.1.429, B.1.427, or B.1.525. (Item 18) The method according to item 11, wherein the compound is probenecid or a pharmaceutically acceptable salt thereof. (Item 19) The method according to any one of items 1 to 18, wherein the compound is in a pharmaceutical composition further comprising a pharmaceutically acceptable carrier and / or excipient. (Item 20) The method according to any one of items 1 to 18, wherein the compound is administered systemically. (Item 21) The method according to any one of items 1 to 18, wherein the compound is administered orally, parenterally, topically, or mucosally. (Item 22) The method according to any one of items 1 to 18, wherein the compound is administered mucosally to the lungs, nasal mucosa, or a combination thereof. (Item 23) The method according to any one of items 1 to 18, wherein the compound is administered in an amount effective in reducing viral replication. (Item 24) The method according to any one of items 1 to 18, wherein the compound is administered in a dose of 10 mg to 1,000 mg or 50 mg to 500 mg, twice daily as needed, for 14 days as needed. (Item 25) The method according to any one of items 1 to 18, wherein the subject is treated by pulse administration. (Item 26) The method according to item 25, wherein the pulse administration comprises one to five bolus doses of the compound ranging from 1,000 mg to 5,000 mg. (Item 27) The method described in item 26, wherein a drug-free day follows the bolus administration. (Item 28) The method according to any one of items 1 to 18, wherein the compound is administered orally or by infusion. (Item 29) The method described in any one of items 1 to 18, wherein the subject is a human. (Item 30) The method according to any one of items 1 to 18, wherein the compound is administered once or twice daily at a dose of 250 mg to 1,000 mg. (Item 31) The method described in any one of items 1 to 18, wherein the compound is administered at a dose of 500 mg twice daily. (Item 32) The method according to any one of items 1 to 18, wherein the compound is administered to the subject for two weeks or longer.

Claims

1. A composition for treating a subject of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, comprising an effective amount of probenecid, its metabolites, its analogues, and a compound selected from any of the above pharmaceutically acceptable salts, wherein the metabolite or analogue thereof Probenecid acylglucuronide, Probenecid β-ether glucuronide, dl-p-(N-propyl-N-2-hydroxypropylsulfamoyl)benzoic acid, p-(N-propyl-N-3-hydroxypropylsulfamoyl)benzoic acid, p-(N-propyl-N-3-propionnitrilosulfamoyl)benzoic acid, p-(N-propyl-N-2-carboxyethylsulfamoyl)benzoic acid, p-(N-propylsulfamoyl)benzoic acid, p-(N,N-pentamethylenesulfamoyl)benzoic acid, p-(N-propyl-N-2-propenylsulfamoyl)benzoic acid, or A composition comprising p-(N-propyl-N-2-oxopropylsulfamoyl)benzoic acid.

2. The composition according to claim 1, wherein the subject has one or more symptoms from among the diseases, disorders, or illnesses related to SARS-CoV-2.

3. The composition according to claim 2, wherein one or more of the symptoms are selected from fever, sinus and / or pulmonary congestion, runny nose or nasal congestion, cough, sneezing, sore throat, body aches, fatigue, shortness of breath, chest tightness, wheezing during exhalation, chills, muscle pain, headache, diarrhea, malaise, nausea, anosmia, skin rash, and combinations thereof.

4. The composition according to claim 1, wherein the subject has COVID-19.

5. The composition according to claim 1, wherein the subject is asymptomatic.

6. The composition according to claim 1, wherein the SARS-CoV-2 comprises a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1 or 2.

7. The composition according to claim 1, wherein the SARS-CoV-2 comprises a spike protein having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO:

5.

8. The composition according to claim 1, wherein the SARS-CoV-2 comprises one or more spike protein mutations in SEQ ID NO: 5, selected from H69, optionally its deletion; V70, optionally its deletion; G142, optionally G142D; K417, optionally K417N or K417T; E484, optionally E484K; F486, optionally F486L; and the N501 mutation, optionally N501Y or N501T.

9. The composition according to claim 1, wherein the SARS-CoV-2 is derived from the B. 1.1.7, B. 1.351, P. 1, B. 1.1.207, B. 1.429, B. 1.427, or B. 1.525 lineage.

10. The composition according to claim 1, wherein the compound is probenecid or a pharmaceutically acceptable salt thereof.

11. A composition for preventing infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in a subject, comprising an effective amount of probenecid, its metabolites, its analogues, and a compound selected from any of the above pharmaceutically acceptable salts, wherein the metabolite or analogue thereof Probenecid acylglucuronide, Probenecid β-ether glucuronide, dl-p-(N-propyl-N-2-hydroxypropylsulfamoyl)benzoic acid, p-(N-propyl-N-3-hydroxypropylsulfamoyl)benzoic acid, p-(N-propyl-N-3-propionnitrilosulfamoyl)benzoic acid, p-(N-propyl-N-2-carboxyethylsulfamoyl)benzoic acid, p-(N-propylsulfamoyl)benzoic acid, p-(N,N-pentamethylenesulfamoyl)benzoic acid, p-(N-propyl-N-2-propenylsulfamoyl)benzoic acid, or A composition comprising p-(N-propyl-N-2-oxopropylsulfamoyl)benzoic acid.

12. The composition according to claim 11, wherein the subject is exposed to SARS-CoV-2.

13. The composition according to claim 11, wherein the subject has been in close contact with a person infected with SARS-CoV-2.

14. The composition according to claim 11, wherein the SARS-CoV-2 comprises a genome encoded by a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1 or 2.

15. The composition according to claim 11, wherein the SARS-CoV-2 comprises a spike protein having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO:

5.

16. The composition according to claim 11, wherein the SARS-CoV-2 comprises one or more mutations in SEQ ID NO: 5, selected from H69, optionally a deletion thereof; V70, optionally a deletion thereof; G142, optionally G142D; K417, optionally K417N or K417T; E484, optionally E484K; F486, optionally F486L; and the N501 mutation, optionally N501Y or N501T.

17. The composition according to claim 11, wherein the SARS-CoV-2 is derived from the B. 1.1.7, B. 1.351, P. 1, B. 1.1.207, B. 1.429, B. 1.427, or B. 1.525 lineage.

18. The composition according to claim 11, wherein the compound is probenecid or a pharmaceutically acceptable salt thereof.

19. The composition according to any one of claims 1 to 18, wherein the compound is in a pharmaceutical composition further comprising a pharmaceutically acceptable carrier and / or excipient.

20. The composition according to any one of claims 1 to 18, characterized in that the composition is administered systemically.

21. The composition according to any one of claims 1 to 18, characterized in that the composition is administered orally, parenterally, topically, or via mucosal administration.

22. The composition according to any one of claims 1 to 18, characterized in that the composition is administered mucosally to the lungs, nasal mucosa, or a combination thereof.

23. The composition according to any one of claims 1 to 18, characterized in that the composition is administered in an amount effective in reducing viral replication.

24. The composition according to any one of claims 1 to 18, characterized in that the compound is administered in a dose of 10 mg to 1,000 mg or 50 mg to 500 mg, twice daily as needed, for 14 days as needed.

25. The composition according to any one of claims 1 to 18, characterized in that the subject is treated by pulse administration.

26. The composition according to claim 25, characterized in that the pulse administration comprises one to five bolus administrations of 1,000 mg to 5,000 mg of the compound.

27. The composition according to claim 26, characterized in that a drug-free period follows the bolus administration.

28. The composition according to any one of claims 1 to 18, characterized in that the composition is administered orally or by infusion.

29. The composition according to any one of claims 1 to 18, wherein the subject is a human.

30. The composition according to any one of claims 1 to 18, characterized in that the compound is administered once or twice a day in a dose of 250 mg to 1,000 mg.

31. The composition according to any one of claims 1 to 18, characterized in that the compound is administered twice a day at a dose of 500 mg.

32. The composition according to any one of claims 1 to 18, characterized in that the composition is administered to the subject for two weeks or longer.

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