WO2022051557A1

WO2022051557A1 - Glycyrrhizin for the treatment or prophylaxis of viral infection

Info

Publication number: WO2022051557A1
Application number: PCT/US2021/048945
Authority: WO
Inventors: Nazlie LATEFI
Original assignee: Applied Biological Laboratories, Inc.
Priority date: 2020-09-02
Filing date: 2021-09-02
Publication date: 2022-03-10

Abstract

Compositions and methods for the treatment or prophylaxis of infection (e.g., influenza, coronavirus, SARS-CoV-2) and diseases associated therewith (e.g., COVID-19) are provided herein. The present disclosure partially based on the discovery that glycyrrhizin has antiviral activity against SARS-CoV-2 virions which may be increased with the combination of certain actives (e.g., SLPI).

Description

GLYCYRRHIZIN FOR THE TREATMENT OR PROPHYLAXIS OF VIRAL INFECTION

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority to and the benefit of U.S. App. No. 63/073,859, filed September 2, 2020, the entire contents of which are hereby incorporated by reference in their entirety.

BACKGROUND

[0002] The control of the spread of infections in a populace is often difficult to achieve. In particular, pathogenic organisms that spread easily may infect a great number of the populace at a fast rate. Furthermore, without appropriate treatments identified, some infections are able to spread rapidly without curtailment by normal means. In particular, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the strain of coronavirus that causes coronavirus disease 2019 (COVID-19), the illness responsible for the COVID-19 pandemic. Treatments for SARS-CoV-2 are severely lacking contributing to the pandemic and many worldwide states of emergency.

[0003] It is therefore an object of this disclosure to provide compositions and methods for treating coronavirus, and in particular SARS-CoV-2 and Coronavirus -229E.

SUMMARY

In accordance with the foregoing objectives and others, the present disclosure provides methods of treatment or prophylaxis of infection with viral infection (e.g., influenza, H1N1, coxsackie virus, coronavirus such as SARS-CoV-2 or Coronavirus 229E)) in a subject, and compositions for use in the methods disclosed herein. Typically, the pharmaceutical compositions of the present disclosure comprise one or more pharmaceutically acceptable carriers, excipients, and/or diluents and glycyrrhizin or, enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing. In particular, topical administration of these compositions are shown herein to achieve an antiviral effect. The presently disclosed administration regimens of glycyrrhizin such as topical administration may allow for lower administration dosages to achieve an antiviral affect as compared to other routes of administration (e.g., oral). Topical administration may avoid toxic effects associated with other modalities of glycyrrhizin administration (e.g., oral administration). For example, glycyrrhizin administration is known to inhibit the enzyme 11-P-hydroxysteroid dehydrogenase enzyme type 2 resulting in a cortisol induced mineralocorticoid effect and a tendency to elevate sodium levels and reduce potassium levels in a subject. Typical administration may result in side effect such as increased blood pressure (e.g., due to its interaction with the kidneys), heart disease, hormone-sensitive conditions, hyperaldosteronism, hypertonia, or hypokalemia. The presently disclosed methods, however, may avoid (or reduce) these side effects while still offering the antiviral activity to the subject in need thereof.

[0004] The administration of the pharmaceutical compositions of the present disclosure may reduce the viral load of an virions (e.g., influenza, coronavirus such as of SARS-CoV-2) and/or prevent an increase in the viral load of virions in the subject. Furthermore, administration as disclosed herein may not be associated with, or have decreased, side effects as compared to other administration mechanisms of glycyrrhizin or, enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing. In some embodiments, the methods may be used for the treatment or prophylaxis of a disease, disorder, or condition associated with SARS- CoV-2 infection such as COVID-19. In some embodiments, the administration of the compositions of the present invention may occur before virus absorption (e.g., as in prophylaxis) to the site of infection and/or during and/or after virus absorption (e.g., as in prophylaxis and/or treatment) to the site of infection in the subject in need thereof. In some methods the administration of the pharmaceutical composition topically coats the oral and/or nasal mucosa (e.g., the pharmaceutical composition is formulated as an oral spray, a nasal spray, or a lozenge). In some embodiments, the pharmaceutical composition is administered daily following infection. In some embodiments, the pharmaceutical composition may be administered each day for at least two days (e.g., from 2-14 days, from 2-10 days, from 2-7 days, from 2-5 days).

[0005] In some embodiments, the compositions of the present disclosure may include compounds of the licorice family (e.g., Glycyrrhiza glabra plant), including glycyrrhetic acid, glycyrrhizic acid, and derivatives thereof. For example, salts and esters may be used to afford the antiviral effects described herein. Extracts of the Glycyrrhiza glabra plant may also be used. Suitable salts of the prior compounds include metal and ammonium salts of these compounds. Suitable esters include unsaturated and saturated C2-C24, or Cio-C24or C16-C24, esters. As specific examples, mention may be made of oil-soluble licorice extract, glycyrrhizic and glycyrrhetic acids, monoammonium glycyrrhizinate, monopotassium glycyrrhizinate, dipotassium glycyrrhizinate, 1-beta-glycyrrhetic acid, stearyl glycyrrhetinate, 3-stearyloxy- glycyrrhetinic acid, and 3-succinyloxy-beta-glycyrrhetinate disodium, or stearyl glycyrrhetinate.

[0006] The method for the treatment of infection (e.g., coronavirus such as SARS-CoV-2) may comprise topical administration of a pharmaceutical composition to a subject in need thereof (e.g., a subject having a viral load of coronavirus such as SARS-CoV-2), wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients and glycyrrhizin (e.g., (3p,20P)-20-carboxy-l l-oxo-30-norolean-12-en-3-yl 2-O-P-D- glucopyranuronosyl-a-D-glucopyranosiduronic acid) or, enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing.

[0007] The method for the prophylaxis of infection (e.g., infection from coronavirus such as SARS-CoV-2) may comprise topical administration of a pharmaceutical composition to a subject in need thereof (e.g., a subject having a viral load of the SARS-CoV-2), wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients and glycyrrhizin (e.g., (3p,20P)-20-carboxy-l l-oxo-30-norolean-12-en-3-yl 2-O-P-D- glucopyranuronosyl-a-D-glucopyranosiduronic acid) or, enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing.

[0008] The present disclosure also includes pharmaceutical compositions for the treatment and/or prophylaxis of viral infection (e.g. coronavirus such as SARS-CoV-2) comprising glycyrrhizin enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing. In some embodiments, the pharmaceutical composition further comprises a protease inhibitor (e.g., serine protease inhibitor) such as elafin or antileukoproteinase (also known as secretory leukocyte protease inhibitor (SLPI)). In some embodiments, the pharmaceutical composition is formulated as an oral spray, a nasal spray, or a lozenge. The pharmaceutical composition may comprise a weight ratio of glycyrrhizin: SLPI of from 100: 1 to 1 : 100 (e.g., 50: 1 to 1 :50, 25: 1 to 1 :25, 20: 1 to 1 :20, 10: 1 to 1 : 10, 5: 1 to 1 :5, 2: 1 to 1 :2, 1 : 1 to 1 : 100, 1 : 1 to 1 :50, 1 :1 to 1 :25, 1 : 1 to 1 :20, 1 : 1 to 1: 10, 1 : 1 to 1 :5, 1 : 1 to 1 :2, 100: 1 to 1 : 1, 50: 1 to 1 : 1, 25: 1 to 1 : 1, 20: 1 to 1 : 1, 10: 1 to 1 : 1, 5: 1 to 1 : 1, 2: 1 to 1 : 1, 20: 1 to 10: 1, 20: 1 to 15: 1, 17: 1 to 16: 1). In various implementations, the pharmaceutical composition comprises less than (or from 0.0001% to) 50% (e.g., less than 40%, less than 30%, less than 20%, less than 10%, from 0.001 to 0.01%, from 0.01% to 0.1%, from 0.1% to 1%, from 1% to 10%, from 10% to 50%) glycyrrhizin by weight of the composition. In various implementations, the pharmaceutical composition comprises less than (or from 0.0001% to) 50% (e.g., less than 40%, less than 30%, less than 20%, less than 10%, from 0.001 to 0.01%, from 0.01% to 0.1%, from 0.1% to 1%, from 1% to 10%, from 10% to 50%) SLPI by weight of the composition. In some embodiments, the pharmaceutical composition for the treatment and/or prophylaxis of viral may comprise a) less than (or from 0.0001% to) 50% (e.g., less than 40%, less than 30%, less than 20%, less than 10%, from 0.001 to 0.01%, from 0.01% to 0.1%, from 0.1% to 1%, from 1% to 10%, from 10% to 50%) glycyrrhizin by weight of the composition; and b) less than (or from 0.0001% to) 50% (e.g., less than 40%, less than 30%, less than 20%, less than 10%, from 0.001 to 0.01%, from 0.01% to 0.1%, from 0.1% to 1%, from 1% to 10%, from 10% to 50%) SLPI by weight of the composition. In certain aspects, the pharmaceutical composition comprises a concentration of glycyrrhizin of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL, from 5 mg/mL to 2400 mg/mL). In some embodiments, the pharmaceutical composition may be formulated to provide a concentration of glycyrrhizin on the area of topical administration (e.g., the oral mucosa, the nasal mucosa) of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL, from 1 mg/mL to 50 mg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 25 mg/mL, from 5 mg/mL to 2400 mg/mL). In certain aspects, the pharmaceutical composition comprises a concentration of SLPI of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL). In some embodiments, the pharmaceutical composition may be formulated to provide a concentration of SLPI on the area of topical administration (e.g., the oral mucosa, the nasal mucosa) of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL, from 1 mg/mL to 50 mg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 25 mg/mL). In some embodiments, the pharmaceutical composition has a concentration of SLPI of greater than 30 pg/mL (e.g., more than 40 pg/mL, more than 50 pg/mL, more than 60 pg/mL, more than 70 pg/mL, more than 80 pg/mL, more than 90 pg/mL, more than 100, pg/mL, more than 150 pg/mL, more than 200 pg/mL, more than 250 pg/mL, more than 275 pg/mL, 30 pg/mL to 5000 mg/mL, 30 pg/mL to 2500 mg/mL, 30 pg/mL to 2000 mg/mL, 30 pg/mL to 1000 mg/mL, 30 pg/mL to 600 mg/mL, 30 pg/mL to 300 mg/mL, 30 pg/mL to 100 mg/mL, 40 pg/mL to 2500 mg/mL, 40 pg/mL to 2000 mg/mL, 40 pg/mL to 1000 mg/mL, 40 pg/mL to 600 mg/mL, 40 pg/mL to 300 mg/mL, 40 pg/mL to 100 mg/mL, 50 pg/mL to 2500 mg/mL, 50 pg/mL to 2000 mg/mL, 50 pg/mL to 1000 mg/mL, 50 pg/mL to 600 mg/mL, 50 pg/mL to 300 mg/mL, 30 pg/mL to 100 mg/mL, 100 pg/mL to 2500 mg/mL, 100 pg/mL to 2000 mg/mL, 100 pg/mL to 1000 mg/mL, 100 pg/mL to 600 mg/mL, 100 pg/mL to 300 mg/mL, 30 pg/mL to 100 mg/mL).

[0009] The pharmaceutical composition may be formulated for delivery to the oral and/or nasal mucosa (e.g., as an oral spray, nasal spray, lozenge), wherein said pharmaceutical composition comprises a pharmaceutically acceptable carrier, excipient, and diluent; and less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 10 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 5 mg/mL) glycyrrhizin enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing; wherein said composition is characterized as resulting in a modulation of the tissue integrity of said oral and/or nasal mucosa following application thereto. In particular embodiments, the pharmaceutical composition, wherein said oral and/or nasal mucosa of the characterization may be infected with virions (e.g., influenza virions, coronavirus virions such as SARS-CoV-2). In some embodiments, the pharmaceutical composition may be further characterized as reducing the number of virions following application of the composition to said mucosa (e.g., as compared to administration of an otherwise identical composition not comprising said glycyrrhizin, for example at from 24 hours to 72 hours following administration).

[0010] The methods of the present disclosure may comprise topical administration is topical administration to the oral or nasal mucosa. In some embodiments, the pharmaceutical composition is an oral spray, a nasal spray, or a lozenge. In various implementations, the pharmaceutical composition comprises less than (or from 0.0001% to) 50% (e.g., less than 40%, less than 30%, less than 20%, less than 10%, from 0.001 to 0.01%, from 0.01% to 0.1%, from 0.1% to 1%, from 1% to 10%, from 10% to 50%) glycyrrhizin by weight of the composition. In certain aspects, the pharmaceutical composition comprises a concentration of glycyrrhizin of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL, from 5 mg/mL to 2400 mg/mL). In some embodiments, the pharmaceutical composition may be formulated to provide a concentration of glycyrrhizin on the area of topical administration (e.g., the oral mucosa, the nasal mucosa) of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL, from 1 mg/mL to 50 mg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 25 mg/mL).

[0011] A method for the treatment and/or prophylaxis of respiratory infection may comprise administering a pharmaceutical composition comprising glycyrrhizin and a pharmaceutically acceptable carrier, excipient, and diluent to the oral and/or nasal mucosa of a subject in need thereof; wherein said pharmaceutical composition comprises less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 10 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 5 mg/mL) glycyrrhizin or enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing; wherein said glycyrrhizin is administered in an amount sufficient to reduce the viral infection in said subject (e.g., at 48 hours following infection or exposure to infection, at 72 hours following infection or exposure to infection).

[0012] In some embodiments, the method may comprise administering a pharmaceutical composition comprising glycyrrhizin and a pharmaceutically acceptable carrier, excipient, and diluent to a subject having a viral infection or a subject having been exposed to a viral infection; wherein said pharmaceutical composition comprises less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL) glycyrrhizin or enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing; and wherein said glycyrrhizin is administered in an amount sufficient to reduce the viral infection in said subject (e.g., at 48 hours following infection or exposure to infection, at 72 hours following infection or exposure to infection).

[0013] Methods are disclosed which may comprise administering a pharmaceutical composition comprising glycyrrhizin and a pharmaceutically acceptable carrier, excipient, and diluent to the oral and/or nasal mucosa of a subject having a viral infection or a subject having been exposed to a viral infection; wherein said pharmaceutical composition comprises less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 10 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 5 mg/mL) glycyrrhizin or enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing; and wherein said glycyrrhizin is administered in an amount sufficient to modulate (e.g., increase, decrease) the tissue integrity of the oral and/or nasal mucosa (e.g., as compared to the tissue of otherwise identical mucosa at, for instance, 48 hours or 72 hours, following initial administration of said composition).

[0014] As shown herein glycyrrhizin and enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing has antiviral activity against infections such as SARS-CoV-2 infection at certain concentrations thereof. In some implementations, the viral load of the SARS-CoV-2 in the subject at 48 hours following administration is less than (or from 0.01%) 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of the compound) . In some aspects, the viral load of the SARS-CoV-2 in the subject at 72 hours is less than (or from 0.01%) 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of the compound). Viral loads may be determined, for example, using polymerase chain reaction (PCR) analysis and quantification.

[0015] In some embodiments, the administration may delivers the glycyrrhizin or enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing to the oral and/or nasal mucosa (e.g., coats the oral and/or nasal mucosa or a portion thereof with a composition comprising glycyrrhizin) and the viral load of the SARS-CoV-2 on the oral/and nasal mucosa at 48 hours following administration is less than (or from 0.01%) 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of the compound). In various implementations, the administration may deliver the glycyrrhizin to the oral and/or nasal mucosa (e.g., coats the oral and/or nasal mucosa or a portion thereof with a composition comprising glycyrrhizin) and the viral load of the SARS-CoV-2 on the oral/and nasal mucosa at 72 hours following administration is less than (or from 0.01%) 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of the compound).

[0016] In some embodiments, the administration is daily administration such as administering the composition once a day or two or more times daily. In some embodiments, the compositions of the present disclosure are administered daily for at least two days (e.g., from 2-30 days or from 2-14 days or from 2-10 days or from 2-8 days or from 2-5 days).

[0017] In some embodiments, glycyrrhizin or enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing is the sole active ingredient in the pharmaceutical composition (i.e., an otherwise identical composition not comprising the sole active agent would have no effect on the viral load as compared to a control). In some embodiments, the pharmaceutical composition comprises an additional antiviral agent. In various implementations, pharmaceutical composition comprises an enzyme (e.g., protease inhibitors such as secretory leukocyte protease inhibitor (SLPI)). In various implementations, the pharmaceutical composition further comprises a glycoprotein (e.g., sialic acid such as siallyllactose, 6’- sialyllactose, 3’-sialyllactose combinations of 6’-sialyllactose and 3-sialyllactose such as in a 10: 1 to 1 : 10 or 5: 1 to 1 :5 or 2: 1 to 1 :2 weight ratio). In some embodiments, the pharmaceutical composition comprises lactoferrin (e.g., human lactoferrin, apolactoferrin) and/or lysozyme (e.g., chicken lysozyme). In various implementations, the pharmaceutical composition does not comprise (or comprises less than (or from 0.01% to) 5% by weight or less than 1% by weight, or from 0.1% by weight) an enzyme (e.g., protease inhibitors such as secretory leukocyte protease inhibitor (SLPI)) and/or a glycoprotein (e.g., sialic acid such as siallyllactose, 6’-sialyllactose, 3’-sialyllactose combinations of 6’-sialyllactose and 3- sialyllactose such as in a 10: 1 to 1 : 10 or 5: 1 to 1 :5 or 2: 1 to 1 :2 weight ratio) and/or comprises lactoferrin (e.g., human lactoferrin, apolactoferrin) and/or lysozyme (e.g., chicken lysozyme).

[0018] The methods of the present disclosure may be used against coronavirus infection, and in particular various strains of SARS-CoV-2 infection. In various implementations, the viral RNA of the SARS-CoV-2 comprise the sequence ORFlb-nspl4 or fragments thereof (e.g., 0RFlb-nspl4F, 0RFlb-nspl4R, 0RFlb-nspl4P). In some embodiments, the viral RNA of the SARS-CoV-2 comprise one or more sequences complimentary to:

5 ’ -TGGGGYTTTACRGGTAACCT-3 ’ ,

5’-AACRCGCTTAACAAAGCACTC-3’, or

5’-TAGTTGTGATGCWATCATGACTAG- 3’.

[0019] In some embodiments, the methods of the present disclosure further comprise obtaining a biological sample from a subject and/or assaying the biological sample for the presence of SARS-CoV-2. The biological sample may be taken from the oral and/or nasal mucosa with a swab. In some embodiments polymerase chain reaction (PCR) analysis is performed on the biological sample to determine the presence of SARS-CoV-2. Suitable assays include those designed by the School of Public Health/University of Hong Kong (Leo Poon, Daniel Chu, and Malik Peiris) and published by the World Health Organization at https://www.who.int/docs/default-source/coronaviruse/peiris-protocol-16-l- 2O.pdf?sfvrsn=aflaac73_4, which is hereby incorporated by reference in its entirety.

BRIEF DESCRIPTION OF FIGURES

[0020] FIG. 1A is a schematic of the protocol used to quantify SARS-CoV-2 infection on MucilAir media as described in Example 1.

[0021] FIG. IB shows various metrics of the protocol described in Example 1.

[0022] FIG. 1C is a table detailing the experimental protocol of Example 1.

[0023] FIG. 2A shows the viral SARS-CoV-2 quantification as determined by RT qPCR in each test condition at 48 hours post inoculation (pi). Statistical comparison was performed daily comparing all conditions to vehicle using Kruskal -Wallis test with Dunn’s posttests (Prism 6.0 GraphPad: *<p<0.05).

[0024] FIG. 2B shows the viral SARS-CoV-2 quantification as determined by RT qPCR in each test condition at 48 hours post inoculation (pi).

[0025] FIG. 3A shows the infectious SARS-CoV-2 titration using TCID50 from the apical wash of MucilAir™ at 48 hours post inoculation (pi). The dotted line indicates the limit of detection (63.2 TCID50/mL). [0026] FIG. 3B shows the infectious SARS-CoV-2 titration using TCID50 from the apical wash of MucilAir™ at 72 hours post inoculation (pi). The dotted line indicates the limit of detection (63.2 TCID50/mL).

[0027] FIG. 4 shows the TEER measurements for each test condition at 48 hours and 72 hours post inoculation (pi). The threshold limit is shown as the dotted line at 100 Q.cm².

[0028] FIG. 5 shows the relative SARS-CoV-2 genome copy number on the apical side of MucilAir Pool with one hour pretreatment (Bl and B2) and repeated apical exposure to test compositions once a day (Bl, B2, and B3) (n=4, 3 cultures) at 48 and 72 hours. Antiviral remdesiver at 5pM was added in basolateral medium at one hour post-inoculation. Statistical comparison was performed comparing all conditions to vehicle using two-way ANOVA with Dunnett’s multiple comparison post-tests (Prism 6.0 GraphPad, *p<0.05, **p<0.05, ***p<0.001, ****p<0.0001). Error bars representing the standard deviation are also provided.

[0029] FIGS. A and 6B show infectious SARS-CoV-2 titration using TCID50 from the apical wash of MucilAir Pool with one hour pretreatment and repeated apical exposure of the Bl and B2 compositions (n=4, 3 cultures) once a day at 48 hours (FIG. 6A) and 72 hours (FIG. 6B). Antiviral remdesivier at 5 pM was added in basolateral medium at one hour post-inoculation. The dotted line represents the limit of detection 63.2 TCID50.mL. Statistical comparison was performed comparing all conditions to vehicle using two-way ANOVA with Dunnett’s multiple comparison post-tests (Prism 6.0 GraphPad, *p<0.05, **p<0.05, ***p<0.001, ****p<0.0001). Error bars representing the standard deviation are also provided.

[0030] FIG. 7 shows the effect of pre-treatment (Bl and B2) and repeated apical exposure to test compositions Bl, B2, and B3 on tissue integrity as measured by TEER from MucilAir Pool upon SARS-CoV-2 infection. TEER was measured before inoculation, and at 2 and 3 days post infection (pi, n=4, 3 cultures). Threshold limit value is lOOQ.cm².

[0031] FIGS. 8 A and 8B provide the measured genome copies/mL from each sample at 24 hours (FIG. 8A) and 48 hours (FIG. 8B). Error bars represent the standard deviation of the measurements.

DETAILED DESCRIPTION

[0032] Detailed embodiments of the present disclosure are disclosed herein; however, it is to be understood that the disclosed embodiments are merely illustrative of the disclosure that may be embodied in various forms. In addition, each of the examples given in connection with the various embodiments of the disclosure is intended to be illustrative, and not restrictive. [0033] All terms used herein are intended to have their ordinary meaning in the art unless otherwise provided. All concentrations are in terms of percentage by weight of the specified component relative to the entire weight of the topical composition, unless otherwise defined.

[0034] As used herein, “a” or “an” shall mean one or more. As used herein when used in conjunction with the word “comprising,” the words “a” or “an” mean one or more than one. As used herein “another” means at least a second or more.

[0035] As used herein, all ranges of numeric values include the endpoints and all possible values disclosed between the disclosed values. The exact values of all half-integral numeric values are also contemplated as specifically disclosed and as limits for all subsets of the disclosed range. For example, a range of from 0.1% to 3% specifically discloses a percentage of 0.1%, 1%, 1.5%, 2.0%, 2.5%, and 3%. Additionally, a range of 0.1 to 3% includes subsets of the original range including from 0.5% to 2.5%, from 1% to 3%, and from 0.1% to 2.5%. It will be understood that the sum of all weight % of individual components will not exceed 100%.

[0036] Throughout this description, various components may be identified having specific values or parameters, however, these items are provided as exemplary embodiments. Indeed, the exemplary embodiments do not limit the various aspects and concepts of the present disclosure as many comparable parameters, sizes, ranges, and/or values may be implemented. Unless otherwise specified, the terms “first,” “second,” and the like, “primary,” “secondary,” and the like, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another.

[0037] Any polypeptide or protein described herein may be a protein or fragment thereof comprising at least 60% or at least 70% or at least 80% at least 85% or at least 90% amino acid sequence identity to the sequence, for example, as identified at NCBI.

[0038] For example, the secretory leukocyte protease gene (SLPI) is a gene which encodes a secreted enzyme inhibitor which may protect epithelial tissues from serine proteases. SLPI protein may be an amino acid sequence containing at least eight cysteine residues and possessing serine protease inhibitor activity, wherein at least one active site comprises one or more amino acid sequences selected from the group consisting of:

Gln-Cys-Leu-R²-Tyr-Lys-Lys-Pro-Glu-Cys-Gln-Ser-Asp; and

Gln-Cys-R⁸-R³-R⁹-Asn-Pro-Pro-Asn-Phe-Cys-Glu-R⁴-Asp wherein R², R³ and R⁴ are the same or different and are independently selected from the group consisting of methionine, valine, alanine, phenylalanine, tyrosine, tryptophan, lysine, glycine and arginine residues; and R⁸ and R⁹ are the same or different and are selected from the group consisting of methionine, valine, alanine, phenylalanine, tyrosine, tryptophan, lysine, glycine, leucine and arginine residues. The SLPI may have the following amino acid sequence:

Ser-Gly-Lys-Ser-Phe-Lys-Ala-Gly-Val-Cys-Pro-Pro-Lys-Lys-Ser-Ala-Gln-Cys- Leu-Arg- Tyr-Lys-Lys-Pro-Glu-Cys-Gln-Ser-Asp-Trp-Gln-Cys-Pro-Gly-Lys-Lys- Arg-Cys-Cys-Pro- Asp-Thr-Cys-Gly-Ile-Lys-Cys-Leu-Asp-Pro-Val-Asp-Thr-Pro- Asn-Pro-Thr-Arg-Arg-Lys- Pro-Gly-Lys-Cys-Pro-Val-Thr-Tyr-Gly-Gn-Cys-Leu- Met-Leu-Asn-Pro-Pro-Asn-Phe-Cys- Glu-Met-Asp-Gly-Gln-Cys-Lys-Arg-Asp-Leu-Lys-Cys-Cys-Met-Gly-Met-Cys-Gly-Lys-Ser- Cys-Val-Ser-Pro-Val- Lys-Ala; or be a fragment thereof having at least 60% or at least 70% or at least 80% at least 85% or at least 90% amino acid sequence identity thereto. In some embodiments, an SLPI analog may be used such as:

R^Gly-Lys-Ser-Phe-Lys-Ala-Gly-Val-Cys-Pro-Pro-Lys-Lys-Ser- Ala-Gln-Cys-Leu-R²-Tyr- Lys-Lys-Pro-Glu-Cys-Gln-Ser-Asp-Trp- Gln-Cys-Pro-Gly-Lys-Lys-Arg-Cys-Cys-Pro-Asp- Thr-Cys-Gly-Ile- Lys-Cys-Leu-Asp-Pro-Val-Asp-Thr-Pro-Asn-Pro-Thr-Arg-Arg-Lys-Pro- Gly-Lys-Cys-Pro-Val-Thr-Tyr-Gly-Gln-Cys-R⁸-R³-R⁹-Asn- Pro-Pro- Asn-Phe-Cys-Glu-R⁴- Asp-Gly-Gln-Cys-Lys-Arg-Asp-Leu- Lys-Cys-Cys-R⁵-Gly-R⁶-Cys-Gly-Lys-Ser-Cys-Val- Ser-Pro-Val- Lys R⁷ wherein R¹ and R⁷ are the same or different and are independently selected from the group consisting of serine, alanine or a substituted amino acid residue;

R², R³, R⁴, and R⁶ are the same or different and are independently selected from the group consisting of methionine, valine, alanine, phenylalanine, tyrosine, tryptophan, lysine, glycine and arginine; and

R⁸ and R⁹ are the same or different and are independently selected from the group consisting of methionine, valine, alanine, phenylalanine, tyrosine, tryptophan, lysine, glycine, leucine and arginine.

[0039] By “consist essentially” it is meant that the ingredients include only the listed components along with the normal impurities present in commercial materials and with any other additives present at levels which do not affect the operation of the disclosure, for instance at levels less than 5% by weight or less than 1% or even 0.5% by weight. [0040] The method for the treatment of SARS-CoV-2 infection may comprise topical administration of a pharmaceutical composition to a subject in need thereof (e.g., a subject having a viral load of the SARS-CoV-2), wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients and glycyrrhizin ((3p,20P)-20-carboxy- 11 -oxo-30-norolean-12-en-3-yl 2-O-P-D-glucopyranuronosyl-a-D-glucopyranosiduronic acid) or, enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing.

[0041] The method for the prophylaxis of SARS-CoV-2 infection may comprise topical administration of a pharmaceutical composition to a subject in need thereof (e.g., a subject having a viral load of the SARS-CoV-2), wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients and glycyrrhizin ((3p,20P)-20-carboxy- 11 -oxo-30-norolean-12-en-3-yl 2-O-P-D-glucopyranuronosyl-a-D-glucopyranosiduronic acid) or, enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing.

[0042] The term “pharmaceutical composition,” as used herein, represents a composition containing a compound described herein formulated with a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gel cap); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other formulation described herein (see below).

[0043] As used herein, the phrase “pharmaceutically acceptable” generally safe for ingestion or contact with biologic tissues at the levels employed. Pharmaceutically acceptable is used interchangeably with physiologically compatible. It will be understood that the pharmaceutical compositions of the disclosure include nutraceutical compositions (e.g., dietary supplements) unless otherwise specified. [0044] Unit dosage forms, also referred to as unitary dosage forms, often denote those forms of medication supplied in a manner that does not require further weighing or measuring to provide the dosage (e.g., tablet, capsule, caplet, lozenge). The compositions of the present disclosure may be present as unit dosage forms. For example, a unit dosage form may refer to a physically discrete unit suitable as a unitary dosage for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with any suitable pharmaceutical excipient or excipients. Exemplary, non-limiting unit dosage forms include a tablet (e.g. , a chewable tablet), caplet, capsule (e.g., a hard capsule or a soft capsule), lozenge, film, strip, and gel cap. In certain embodiments, the compounds described herein, including crystallized forms, polymorphs, and solvates thereof, may be present in a unit dosage form.

[0045] Useful pharmaceutical carriers, excipients, and diluents for the preparation of the compositions hereof, can be solids, liquids, or gases. These include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The pharmaceutically acceptable carrier or excipient does not destroy the pharmacological activity of the disclosed compound and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the compound. Thus, the compositions can take the form of tablets, pills, capsules, suppositories, powders, enterically coated or other protected formulations (e.g., binding on ion-exchange resins or packaging in lipid-protein vesicles), sustained release formulations, solutions, suspensions, elixirs, and aerosols. The carrier can be selected from the various oils including those of petroleum, animal, vegetable or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, and sesame oil. Water, saline, aqueous dextrose, and glycols are examples of liquid carriers, particularly (when isotonic with the blood) for injectable solutions. For example, formulations for intravenous administration comprise sterile aqueous solutions of the active ingredient(s) which are prepared by dissolving solid active ingredient(s) in water to produce an aqueous solution and rendering the solution sterile. Suitable pharmaceutical excipients include starch, cellulose, chitosan, talc, glucose, lactose, gelatin, malt, rice, flour, chalk, silica, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk, glycerol, propylene glycol, water, and ethanol. The compositions may be subjected to conventional pharmaceutical additives such as preservatives, stabilizing agents, wetting or emulsifying agents, salts for adjusting osmotic pressure, and buffers. Suitable pharmaceutical carriers, excipients, diluents, and formulations are described in Remington’s Pharmaceutical Sciences by E. W. Martin, which is hereby incorporated by reference in its entirety. Such compositions will, in any event, contain an effective amount of the active compound together with a suitable carrier so as to prepare the proper dosage form for administration to the recipient.

[0046] Non-limiting examples of pharmaceutically acceptable carriers and excipients include sugars such as lactose, glucose and sucrose; starches such as com starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as polyethylene glycol and propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate; coloring agents; releasing agents; coating agents; sweetening, flavoring and perfuming agents; preservatives; antioxidants; ion exchangers; alumina; aluminum stearate; lecithin; self-emulsifying drug delivery systems (SEDDS) such as d-atocopherol polyethyleneglycol 1000 succinate; surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices; serum proteins such as human serum albumin; glycine; sorbic acid; potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts; colloidal silica; magnesium trisilicate; polyvinyl pyrrolidone; cellulose-based substances; polyacrylates; waxes; and polyethylene-polyoxypropylene-block polymers. Cyclodextrins such as a-, P-, and y-cyclodextrin, or chemically modified derivatives such as hydroxyalkylcyclodextrins, including 2-and 3 -hydroxypropyl -cyclodextrins, or other solubilized derivatives can also be used to enhance delivery of the compounds described herein.

[0047] The pharmaceutically acceptable carrier may be selected to provide a specified residence time in the mucosa of a subject. In some embodiments, the “residence time” of the inventive compositions on the mucosa represent average residence times from studies involving multiple applications (intranasal and/or oral) using a sample of multiple individuals sufficient to approximate the population at large. In some embodiments, at least 25% (and preferably, at least 30%, or at least 40% or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%) by weight of the initially applied active ingredients remain on the mucosa after the specified duration of time (e.g., from 1 minute to 60 minutes, from 5 minutes to 40 minutes, from ten minutes to 35 minutes). In some embodiments, the pharmaceutically acceptable carrier at 25°C has the Hansen Solubility Parameters of energy from dispersion (5d), energy from dipolar intermolecular force between molecules (6_P), energy from hydrogen bonds (6h) of from 15 to 18, from 12 to 15, from 21 to 25, respectively.

[0048] In some embodiments, the pharmaceutical composition comprises one or more ingredients such as polymers, thickening agents, viscosity adjusters, release modifying agents (e.g., hydrophilic and/or hydrophobic polymers) to control the residence time of glycyrrhizin on the oral and/or nasal mucosa.

[0049] The pharmaceutically acceptable carrier may be aqueous. In some embodiments, the pharmaceutically acceptable carrier is free of mercurial preservatives. The solvent may be 1,2- propanediol, 1,3 -propanediol and a variety of aqueous carriers can be used, e.g. buffered water, 0.9 percent saline, buffered aqueous-ethanol solutions and the like. Combinations of any of these carriers are within the scope of the invention. These compositions can be sterilized by conventional, well-known sterilization techniques, or can be sterile filtered. The resulting solutions can be packaged for use as is or mixed as an adjuvant to another medication. A composition can contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents, taste modifiers, sweeteners, wetting agents and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, or triethanolamine oleate. In some embodiments, the pharmaceutically acceptable carrier is a mixture of water and a polyol. In some embodiments, the pharmaceutically acceptable carrier is a mixture of water and propanediol (e.g. 1,2-propendediol, 1,3- propanediol). In some embodiments, the pharmaceutical composition is a mixture of water and glycerin. The pharmaceutically acceptable carrier may be from 1%— 35% (e.g. 5% -30%) aqueous solution of propanediol or glycerin by weight of the aqueous carrier. Some pharmaceutically acceptable carriers include 20% aqueous solution of 1,3 -propanediol, 20% aqueous solution of glycerin, 10% aqueous solution of 1,3-propanediol, 10% aqueous solution of glycerin, 20% aqueous solution of 1,3-propanediol with 1% sunflower oil and 5% polysorbate 80, 20% aqueous solution of glycerin with 1% sunflower oil and 5% polysorbate 80, 10% aqueous solution of 1,3-propanediol with 1% sunflower oil and 5% polysorbate 80, 10% aqueous solution of glycerin with 1% sunflower oil and 5% polysorbate 80, the Versaflex V-175 polymeric emulsifier system (i.e. sucrose palmitate, glyceryl stearate, glyceryl sterate citrate, sucrose, mannan, and Xanthan gum), the Versaflex V-175 polymeric emulsifier system with 3% sunflower oil, the Versaflex V-175 polymeric emulsifier system with 3% sunflower oil and from 5 to 30% propanediol or glycerin, the Versaflex V-175 emulsifier system with 3% acetylated monoglyceride, and the Versaflex V-175 emulsifier system with 3% acetylated monoglyceride and from 5 to 30% propanediol or glycerin.

[0050] Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, or magnesium carbonate. Examples of suitable pharmaceutical carriers are described in “Remington's Pharmaceutical Sciences” by E. W. Martin, the contents of which is hereby incorporated by reference in its entirety. Such compositions will generally contain a therapeutically effective amount of the therapeutic agent and/or the immunotherapeutic agent, in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

[0051] Tonicity control agents may be present in the pharmaceutical composition as one of the components therein. These agents may adjust the tonicity of the formulation, and/or adjust the osmolarity of the formulation. The tonicity agents may be added in amount to adjust the tonicity of a dissolved formulation (e.g., a lozenge such that when dissolved has a specified tonicity). Suitable tonicity agents include salts such as sodium chloride or sugars. The tonicity may be adjusted to a desired tonicity for the therapeutic use. For example, a desired tonicity may be one that enhances intranasal mucosal delivery while minimizing discomfort of delivery to the nasal cavity. The tonicity of the formulation may be isotonic, hypotonic, or hypertonic. The osmolality may be adjusted to a desired osmolality. A desired osmolality for certain compositions (e.g., nasal sprays, oral sprays, lozenges) may be between 290 mOsm/kg and 500 mosm/kg. In some embodiments, the formulation is isoosmolar, e.g., 280 mOsm/kg. In some embodiments, the composition is hypoosmotic (e.g., less than 50 mOsm/kg). In some embodiments, the composition is hyperosmotic (e.g, greater than 900 mOsm/kg). [0052] The viscosity of the formulation may be adjusted to for a variety of delivery mechanisms such as increasing time in the on the oral and/or nasal mucosa, altering the spray characteristics, and the like. Viscosity adjusting agents may be included in order to manage factors such as thinning and thixotropic behavior and are key elements in the performance of the dispensed product such as drop particle size, spray angles and also influence the residence time of the product once delivered, for example, in the nasal cavity. The viscosity of the formulation may be for example from 0.1 cp to 500 cp (e.g., from 0.1 cp to 1 cp, from 1 cp to 10 cp, from 10 cp to 100 cp, from 1 cp to 50 cp, from 50 cp to 100 cp, from 100 cp to 150 cp, from 150 cp to 200 cp, from 100 cp to 200 cp, from 200 cp to 250 cp, from 250 cp to 300 cp, from 200 cp to 300 cp, from 300 cp to 350 cp, from 350 cp to 400 cp, from 300 cp to 400 cp, from 400 cp to 450 cp, from 450 cp to 500 cp from 400 cp to 500 cp). Viscosity of the formulation may be measured using a Brookfield DV-IIC Pro viscometer (Brookfield Engineering Laboratories, Middleboro, MA, USA). Briefly, a formulation may be loaded onto the small sample adapter of the viscometer with a constant volume of 5 mL and rested for 30 min with the solvent cap on prior to measurement. An appropriate spindle (e.g., number #18) may be immersed in the test liquid and rotated at a speed of 100 rpm (corresponding to a shear rate of 132 s-1) and yield measurements between 10% and 100% of viscometer torque scale at 25 °C.

[0053] The pharmaceutical composition may comprise a pharmaceutical composition comprising one or more pharmaceutically acceptable excipients, carriers or diluents; and a therapeutically effective amount of an anti-viral agent capable of reducing the infection load in the nasal cavity; wherein the pharmaceutical composition is formulated as an oral or nasal spray. In some embodiments, the composition is formulated for aerosol delivery with a particle size (e.g., as measured by dynamic light scattering) of greater than 1 pm or greater than 5 pm or from 1 pm to 100 pm or from 5 pm to 100 pm or from 10 pm to 100 pm or from 10 pm to 30 pm.

[0054] The glycyrrhizin may be present in the pharmaceutical compositions of the present disclosure as a pharmaceutically acceptable salt. Typically, salts are composed of a related number of cations and anions (at least one of which is formed from the compounds described herein) coupled together (e.g., the pairs may be bonded ionically) such that the salt is electrically neutral. Pharmaceutically acceptable salts may retain or have similar activity to the parent compound (e.g., an ED50 within 10%) and have a toxicity profile within a range that affords utility in pharmaceutical compositions. For example, pharmaceutically acceptable salts may be suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66: 1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. Salts may be prepared from pharmaceutically acceptable non-toxic acids and bases including inorganic and organic acids and bases. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, dichloroacetate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glutamate, glycerophosphate, hemi sulfate, heptonate, hexanoate, hippurate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, isethionate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, methanesulfonate, mucate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pantothenate, pectinate, persulfate, 3 -phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and valerate salts. Representative basic salts include alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium, aluminum salts, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, caffeine, and ethylamine. One particular pharmaceutically acceptable salt of glycyrrhizin is glycyrrhizin ammonium.

[0055] Pharmaceutically acceptable acid addition salts of the disclosure can be formed by the reaction of a compound of the disclosure with an equimolar or excess amount of acid. Alternatively, hemi-salts can be formed by the reaction of a compound of the disclosure with the desired acid in a 2: 1 ratio, compound to acid. The reactants are generally combined in a mutual solvent such as diethyl ether, tetrahydrofuran, methanol, ethanol, /.w-propanol, benzene, or the like. The salts normally precipitate out of solution within, e.g., one hour to ten days and can be isolated by filtration or other conventional methods.

[0056] Solvates of the compounds described herein may the aggregate of the compound or an ion of the compound with one or more solvents. Such solvents may not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, MeOH, EtOH, and AcOH. Solvates wherein water is the solvent molecule are typically referred to as hydrates. Hydrates include compositions containing stoichiometric amounts of water, as well as compositions containing variable amounts of water.

[0057] The term “effective amount” or “therapeutically effective amount” of an agent (e.g glycyrrhizin), as used herein, is that amount sufficient to effect beneficial or desired results, such as clinical results, and, as such, an “effective amount” depends upon the context in which it is being applied. In some embodiments, the compounds are administered in an effective amount for the treatment or prophylaxis of a disease disorder or condition associated with coronavirus infection. In some embodiments, the effective amount may reduce the viral load of infection in the subject in need thereof. In certain implementations, the effective amount may prevent an increase in the viral load of infection in the subject in need thereof. In another embodiment, an effective amount of glycyrrhizin is, for example, an amount sufficient to achieve alleviation or amelioration or prevention or prophylaxis of one or more symptoms or conditions; diminishment of extent of disease, disorder, or condition; stabilized (i.e., not worsening) state of disease, disorder, or condition; preventing spread of disease, disorder, or condition; delay or slowing the progress of the disease, disorder, or condition; amelioration or palliation of the disease, disorder, or condition (e.g., those associated with infection such as COVID-19); and remission (whether partial or total), whether detectable or undetectable, as compared to the response obtained without administration of the agent.

[0058] Typically, the treatment of a disease, disorder, or condition (e.g, the conditions described herein such as those associated with infection such as COVID-19) is an approach for obtaining beneficial or desired results, such as clinical results. Beneficial or desired results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions; diminishment of extent of disease, disorder, or condition; stabilized (i.e., not worsening) state of disease, disorder, or condition; preventing spread of disease, disorder, or condition; delay or slowing the progress of the disease, disorder, or condition; amelioration or palliation of the disease, disorder, or condition; and remission (whether partial or total), whether detectable or undetectable. “Palliating” a disease, disorder, or condition means that the extent and/or undesirable clinical manifestations of the disease, disorder, or condition are lessened and/or time course of the progression is slowed or lengthened, as compared to the extent or time course in the absence of treatment.

[0059] As used herein, the term “subject” refers to any organism to which a composition and/or compound in accordance with the disclosure may be administered, e.g, for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include any animal (e.g, mammals such as mice, rats, rabbits, non-human primates, and humans). A subject in need thereof is typically a subject for whom it is desirable to treat a disease, disorder, or condition as described herein. For example, a subject in need thereof may seek or be in need of treatment, require treatment, be receiving treatment, may be receiving treatment in the future, or a human or animal that is under care by a trained professional for a particular disease, disorder, or condition.

[0060] In some embodiments, the compositions of the present disclosure may be used immediately prior to a high-risk exposure (e.g., boarding an enclosed environment such as a subway, bus, or airplane, entering a hospital). In some embodiments, administration may occur immediately following suspected contact with SARS-CoV-2 (e.g., airborne SARS-CoV-2).

[0061] The methods of the present disclosure may comprise topical administration is topical administration to the oral or nasal mucosa. In some embodiments, the pharmaceutical composition is an oral spray, a nasal spray, or a lozenge. In various implementations, the pharmaceutical composition comprises less than 50% (e.g., less than 40%, less than 30%, less than 20%, less than 10%, from 0.001 to 0.01%, from 0.01% to 0.1%, from 0.1% to 1%, from 1% to 10%, from 10% to 50%) glycyrrhizin enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing by weight of the composition. In certain aspects, the pharmaceutical composition comprises a concentration of glycyrrhizin enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing of less than 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL) and a volume of, for example, from 0.1 mL to 100 mL or from 0.1 mL to 50 mL or from 0.1 mL to 20 mL or from 0.1 mL to 10 mL or from 1 to 100 mL or from 1 to 50 mL or from 1 to 20 mL. In some embodiments, the pharmaceutical composition may be formulated to provide a concentration of glycyrrhizin enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing on the area of topical administration (e.g., the oral mucosa, the nasal mucosa) of less than 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL, from 1 mg/mL to 50 mg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 25 mg/mL).

[0062] For use in administration with oral and/or nasal sprays, the pharmaceutical composition may be formulated to deliver from 20 pL to 1000 pL (e.g., from 20 to 50 pL, from 50 to 100 pL from 100 pL to 150 pL, from 150 pL to 200 pL, from 200 pL to 250 pL, from 250 pL to 300 pL from 300 pL to 350 pL, from 350 pL to 400 pL, from 400 pL to 450 pL, from 450 pL to 500 pL, from 50 pL to 350 pL, from 500 pL to 600 pL from 600 pL to 700 pL, from 700 pL to 800 pL, from 800 pL to 900 pL, from 900 pL to 1000 pL) of the pharmaceutical composition in each spray. The oral and/or nasal spray may be formulated to deliver a narrow spray pattern. In various embodiments the volume per spray may be varied. For example, the volume per spray may be between 50 pL and 150 pL may be. Many different such metered drug pump designs can be adapted for use in administration of the oral and/or nasal sprays including those disclosed in U.S. Pat. Nos. 4,860,738, 4,944,429, 6,321.942, 6,446,839, 6,705.493, 6,708,846, 6,772.915, and 7,182,226, each of which is incorporated by reference in their entirety.

[0063] Each administration (e.g., unit dosage form, spray of an oral and/or nasal spray) of the pharmaceutical composition may include an amount of glycyrrhizin enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing of from 1 pg to 50 mg (e.g., from 1 pg to 10 pg, from 10 pg to 100 pg, from 100 pg to 200 pg, from 200 pg to 300 pg, from 300 pg to 400 pg, from 400 pg to 500 pg, from 500 pg to 600 pg, from 700 pg to 800 pg, from 800 pg to 900 pg, from 900 pg to 1000 pg, from 1 mg to 10 mg, from 10 mg to 20 mg, from 20 mg to 30 mg, from 30 mg to 40 mg, from 40 mg, to 50 mg). In particular embodiments, the composition is formulated to provide a concentration of glycyrrhizin at the delivery are (e.g., the oral and/or nasal mucosa) of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL).

[0064] As shown herein glycyrrhizin has antiviral activity against Coranovirus infection including SARS-CoV-2 and Coronavirus -229E infection. In some implementations, the viral load of the SARS-CoV-2 in the subject at 48 hours following administration is less than 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of the compound) . In some aspects, the viral load of the SARS-CoV-2 in the subject at 72 hours is less than 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of the compound). Viral loads may be determined, for example, using polymerase chain reaction (PCR) analysis and quantification.

[0065] In some embodiments, the administration is daily administration or occurs two or more times daily (e.g., two times daily, three times daily, four times daily).

[0066] In some embodiments, glycyrrhizin or enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing is the sole active ingredient in the pharmaceutical composition (i.e., an otherwise identical composition not comprising the sole active agent would have no effect on the viral load as compared to a control). In some embodiments, the pharmaceutical composition comprises an additional antiviral agent. For example, the weight ratio of glycyrrhizin to antiviral agent may be from 100:1 to 1:100 (e.g., 50:1 to 1:50, 25:1 to 1:25, 20:1 to 1:20, 10:1 to 1:10, 5:1 to 1:5, 2:1 to 1:2, 1:1 to 1:100, 1:1 to 1:50, 1:1 to 1:25, 1:1 to 1:20, 1:1 to 1:10, 1:1 to 1:5, 1:1 to 1:2, 100:1 to 1:1, 50:1 to 1:1, 25:1 to 1:1, 20:1 to 1:1, 10:1 to 1:1, 5:1 to 1:1, 2:1 to 1:1, 20:1 to 10:1, 20:1 to 15:1, 17:1 to 16:1). In various implementations, pharmaceutical composition comprises an enzyme (e.g., protease inhibitors such as secretory leukocyte protease inhibitor (SLPI)). In various implementations, the pharmaceutical composition further comprises a glycoprotein (e.g., sialic acid such as siallyllactose, 6’-sialyllactose, 3’-sialyllactose combinations of 6’-sialyllactose and 3- sialyllactose such as in a 10:1 to 1:10 or 5:1 to 1:5 or 2:1 to 1:2 weight ratio) or a fragment thereof such as a fragment having more than 50% or more than 60% or more than 70% or more than 80% or more than 85% or more than 90% sequence identity to the glycoprotein. In some embodiments, the pharmaceutical composition comprises lactoferrin (e.g., human lactoferrin, apolactoferrin) and/or lysozyme (e.g., chicken lysozyme). In various implementations, the pharmaceutical composition does not comprise an enzyme (e.g., protease inhibitors such as secretory leukocyte protease inhibitor (SLPI)) and/or a glycoprotein (e.g, sialic acid such as siallyllactose, 6’-sialyllactose, 3’-sialyllactose combinations of 6’-sialyllactose and 3- sialyllactose such as in a 10: 1 to 1 : 10 or 5: 1 to 1 :5 or 2: 1 to 1 :2 weight ratio) and/or comprises lactoferrin (e.g., human lactoferrin, apolactoferrin) and/or lysozyme (e.g., chicken lysozyme).

[0067] The methods of the present disclosure may be used against coronavirus infection, and in particular various strains of SARS-CoV-2 infection. In various implementations, the viral RNA of the SARS-CoV-2 comprise the sequence ORFlb-nspl4 or fragments thereof (e.g., ORFlb-nspl4F, ORFlb-nspl4R, ORFlb-nspl4P). In some embodiments, the viral RNA of the SARS-CoV-2 comprise one or more sequences complimentary to:

5 ’ -TGGGGYTTTACRGGTAACCT-3 ’ ,

5’-AACRCGCTTAACAAAGCACTC-3’, or

5’-TAGTTGTGATGCWATCATGACTAG- 3’.

[0068] In some embodiments, the methods of the present disclosure further comprise obtaining a biological sample from a subject and/or assaying the biological sample for the presence of SARS-CoV-2. The biological sample may be taken from the oral and/or nasal mucosa with a swab. In some embodiments polymerase chain reaction (PCR) analysis is performed on the biological sample to determine the presence of SARS-CoV-2. Suitable assays include those designed by the School of Public Health/University of Hong Kong (Leo Poon, Daniel Chu, and Malik Peiris) and published by the World Health Organization at https://www.who.int/docs/default-source/coronaviruse/peiris-protocol-16-l- 2O.pdf?sfvrsn=aflaac73_4, which is hereby incorporated by reference in its entirety.

[0069] Glycyrrhizin, also called glycyrrhizic acid, is a triterpenoid saponin which may be isolated from the roots (Glycyrrhizae Radix) of the plants Glycyrrhiza glabra and G. uralensis Fisch and G. inflata Bat (used in the Chinese Pharmacopoeia). Glycyrrhizin may be obtained from resources that exist in nature such as licorice having glycyrrhizin as one of its components or licorice powder, licorice extract such as licorice root extract. The glycyrrhizin may be acquired from, for example, Minophagen Pharmaceutical Co., Ltd. Alternatively, glycyrrhizin and methods of its production and extraction are described in, for example, Japanese Unexamined Patent Application, First Publication No. Sho 63-2959; Chem. Pharm. Bull. 34, 897 (1986); and Jpn. J. Pharmacol., 71, 281 (1996), each of which are hereby incorporated by reference in their entirety. In some embodiments, the pharmaceutical composition comprises an extract of licorice (e.g., a licorice root extract such as an aqueous and/or alcoholic extract including licorice extracts where the extraction solvent is selected from water, lower alcohols including C1-C4 alcohols, and combinations thereof). In some embodiments the glycyrrhizin is a glycyrrhizinate salt such as dipotassium glycyrrhizinate. In certain embodiments, the glycyrrhizin is dipotassium glycyrrhizinate hydrate.

[0070] In some embodiments, the pharmaceutical composition (including pharmaceutical composition for administration to the oral and/or nasal mucosa) may further comprise a viscosity enhancing agent. In some embodiments, the viscosity enhancing agent includes methylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose and smart hydrogel. In some embodiments, the viscosity enhancing agent is hydroxyethylcellulose. In some embodiments, the intranasal pharmaceutical composition comprises 0.01-1.0% (w/v) hydroxyethylcellulose. In other embodiments, the intranasal pharmaceutical composition comprises 0.05% (w/v) hydroxyethyl cellulose.

[0071] In some embodiments, the pH of the pharmaceutical composition is from 3 to 10 or from 4.0 to 7.5. In other embodiments, the pH of the pharmaceutical composition is from 4.0 to 6.5. In another embodiment the pharmaceutical composition has a pH of from 5.5 to 6.5. In further embodiments, the pharmaceutical composition has a pH of from 6.0 to 6.5. In various implementations, the pH of said aqueous solution or liquid formulation is from pH 3 to pH 7, from pH 3 to pH 6, from pH 4 to pH 6, or from pH 5 to pH 6. These pH ranges may be achieved through the incorporation of one or more pH modifying agents, buffers, and the like. In some embodiments, a pH modifier such as acetic acid, is present in a final concentration of at least 0.001%, preferably at least 0.01%, more preferably between 0.01%-0.2% by weight of the composition.

[0072] In terms of their form, compositions of this invention may include solutions, emulsions (including microemulsions), suspensions, creams, lotions, gels, powders, or other typical solid or liquid compositions used for application to skin and other tissues where the compositions may be used. Such compositions may contain: additional antimicrobials, moisturizers and hydration agents, penetration agents, preservatives, emulsifiers, natural or synthetic oils, solvents, surfactants, detergents, gelling agents, emollients, antioxidants, fragrances, fillers, thickeners, waxes, odor absorbers, dyestuffs, coloring agents, powders, viscosity-controlling agents and water, and optionally including anesthetics, anti-itch actives, botanical extracts, conditioning agents, darkening or lightening agents, glitter, humectants, mica, minerals, polyphenols, silicones or derivatives thereof, sunblocks, vitamins, and phytomedicinals. In certain embodiments, the composition of the invention is formulated with the above ingredients so as to be stable for a long period of time, as may be beneficial where continual or long-term treatment is intended.

[0073] In order to treat, prevent, or prevent recurrence of diseases, disorders, or conditions as discussed herein, the composition of the present disclosure may be administered at least once a day for at least one week. In various embodiments, the composition is administered at least twice a day for at least two days. In certain embodiments, the composition is administered approximately daily, at least daily, twice a week, weekly, or for one month. In certain embodiments, the composition of the invention is administered for several months, such as at least two months, six months, or one year or longer. The invention is further suited for longterm use, which may be particularly beneficial for preventing recurring infection, or for preventing infection or conditions in at-risk or susceptible patients, including immune compromised patients. Such long-term use may involve treatment for at least two years, three years, four years, or even five or more years.

[0074] In another aspect, the composition of the invention is a kit, which contains the compositions of the present disclosure packaged to facilitate dispensing and/or applying the composition to affected or susceptible regions (e.g., nasal and/or oral mucosa, infected nasal and/or oral mucosa). The packaging or dispenser may include a bottle, tube, spray bottle, or other dispenser. In certain embodiments of the invention, the composition is packaged in a concentrated form, and diluted to a desired concentration upon use by the end user. Typically, in these aspects, the composition may be formulated and packaged in a manner suitable for long-term storage to maintain efficacy of the composition.

[0075] The kit may further include additional components to facilitate application of the composition to the affected area, such as, for example, a brush, sponge, cotton swab, or the like.

[0076] The compositions of the present disclosure may be formulated for delivery to the nasal cavity in a form selected from the group consisting of: a powder, a granule; a cachet; a capsule; a tablet; a paste; a cream; a gel; an ointment; a salve; a foam; a paste; a lotion; a cream; an oil suspension; a spray; a suspension; a solution; an emulsion; a patch; a stick; a spray, preferably a nasal spray, or a buccal spray; a mouth wash; an aerosol, from a Venturi effect; and a drink. The compositions may be formulated as a solution, gel, lotion, suspension, cream, ointment, or other formulation appropriate for local application for the treatment or prevention of viral infections (e.g., coronavirus such as SARS-CoV-2 and Coronavirus -229E) and diseases associated therewith such as COVID-19.

[0077] In some embodiments, the compositions of the present disclosure may modulate cytokines such as increasing inflammatory cytokines and/or decreasing inflammatory cytokines in the site of delivery (e.g., mucosa such as the oral or nasal mucosa). Without wishing to be bound by theory, glycchirizin and salts thereof (e.g., dipotassium glycyrrhizinate) may show an effect for promoting activity on an infected surfaces having increased cytokines and infected surfaces having decreased inflammatory cytokines.

[0078] In some embodiments, the pharmaceutical composition is in the form of an oral spray and/or nasal spray. Pharmaceutical products for oral administration are provided. In some embodiments, the pharmaceutical product may comprise:

(a) a body configured to be inserted into a nasal and/or oral passage for dispensing a nasal spray and/or oral spray composition through an orifice;

(b) a reservoir in fluid communication with said orifice, wherein said nasal spray and/or oral spray pharmaceutical composition is contained in said reservoir;

(c) a pump mechanism capable of atomizing and expelling said nasal spray and/or oral spray composition through said orifice in appropriately sized aerosolized droplets; capable of coating the nasal and/or oral mucosa of a user; wherein said nasal spray and/or oral spray composition comprises glycyrrhizin and one or more pharmaceutically acceptable carriers, excipients, and/or diluents.

[0079] These devices may be constructed of material to warrant proper mechanical function and minimize likelihood of chemical interactions with the pharmaceutical composition contained therein. For example, the devices may comprise polyethylene, polypropylene, polyoxy methane, rubber elastomers, stainless steel, aluminum, glass, or combinations thereof.

[0080] Each spray of the oral and/or nasal spray may include an amount of glycyrrhizin of from 1 pg to 50 mg (e.g., from 1 pg to 10 pg, from 10 pg to 100 pg, from 100 pg to 200 pg, from 200 pg to 300 pg, from 300 pg to 400 pg, from 400 pg to 500 pg, from 500 pg to 600 pg, from 700 pg to 800 pg, from 800 pg to 900 pg, from 900 pg to 1000 pg, from 1 mg to 10 mg, from 10 mg to 20 mg, from 20 mg to 30 mg, from 30 mg to 40 mg, from 40 mg, to 50 mg). In particular embodiments, the composition is formulated to provide a concentration of glycyrrhizin at the delivery are (e.g., the oral and/or nasal mucosa) of less than (or from 0.1 mg/mL to) 2400 mg/mL e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL).

[0081] The compounds of the present disclosure (e.g., glycyrrhizin, dipotassium glycyrrhizinate, SLPI) or pharmaceutically acceptable salts, N-oxides, metal complexes, or prodrugs thereof may be used for the treatment or prophylaxis of infection such as infection by coronavirus including SARS-CoV-2 virions. These compounds may also be used in the preparation of a medicament such as a lozenge, a nasal spray, or oral spray for the treatment of prophylaxis of infection such as infection by coronavirus including SARS-CoV-2 virions and the like.

[0082] Methods for the treatment and/or prophylaxis of viral infection (e.g., coronavirus infection, SARS-CoV-2 infection, Coronavirus -229E infection) on a surface (e.g., the oral mucosa in a subject in need thereof, the nasal mucosa in a subject in need thereof, infected MucilAir™ media) are provided comprising applying the compositions of the present disclosure (e.g., a pharmaceutical composition comprising glycyrrhizin and optional SLPI) thereto. The compositions may be applied prior to infection (e.g., prophylactically) and/or after infection (e.g., prophylactically, therapeutically). In some embodiments, the compositions are applied daily. Application of the compositions of the present disclosure may result in a relative SARS-CoV-2 viral infection, as compared to prior to infection, of more than 2 or more than 3 (e.g., 2-10, 3-10, 2-5, 3-5) Mean loglO reduction at 48 hours following the initial application. In some embodiments, application of the compositions of the present disclosure may result in a relative SARS-CoV-2 viral infection, as compared to prior to infection, of more than 2 or more than 3 or more than 4 or more than 5 (e.g., 2-10, 3-10, 4-10, 5-10, 6-10, 2-7, 5-7) Mean log 10 reduction at 72 hours following the initial application. In certain implementations, application of the compositions of the present disclosure may result in a reduction of TCID50/mL of less than 4 or less than 3 or less than 2 loglO TCID50/mL (e.g., 0.1-4, 0.1-3, 0.1-2) at 48 hours following initial application. In some embodiments, application of the compositions of the present disclosure may result in a reduction of TCID50/mL of less than 4 or less than 3 or less than 2 loglO TCID50/mL (e.g., 0.1-4, 0.1-3, 0.1-2) at 48 hours following initial application. In some embodiments, the number of dead viral cells at 48 hours following application and/or at 72 hours after application (e.g., of a composition comprising glycyrrhizin and SLPI) is less than (or from 0.1 to) 70 TCID50/mL or less than 65 TCID50/mL or less than 63.2 TCID50/mL. In some embodiments, some embodiments, the health and/or integrity of the mucosa is modulated by administration of the compositions of the present disclosure (e.g., as measured by TEER measurements). For example, the oral and/or nasal mucosa of a subject may have an increase in TEER (e.g., of less than 10% or less than 20% or less than 25% or less than 30% or from 10% to 30%) as compared to a positive control (e.g., an otherwise identical infected tissue without said composition having been administered). The increase may be measured, for example, at 48 hours post infection (or exposure to infection) or at 72 hours post infection (or exposure to infection). In certain embodiments, the oral and/or nasal mucosa of a subject may have a decrease in TEER (e.g., of less than 10% or less than 20% or less than 25% or less than 30% or from 10% to 30%) as compared to a positive control (e.g., an otherwise identical infected tissue without said composition having been administered). The increase may be measured, for example, at 48 hours post infection (or exposure to infection) or at 72 hours post infection (or exposure to infection). Repeated measurements may be taken such the modulation is demonstrated to be statistically significant (e.g., p<0.05).

EXAMPLES

[0083] Example 1

[0084] SARS-CoV-2 infection after administration of various test compositions was monitored using a 3D model of human airway epithelium, constituted with primary human epithelial cells freshly isolated from nasal, tracheal or bronchial biopsies (MucilAir™). MucilAir™ is composed of basal cells, ciliated cells and mucus cells from the respiratory tract. MucilAir™ is a pseudostratified and ready-to-use 3D model of human airway epithelium, constituted with primary human epithelial cells freshly isolated from nasal, tracheal or bronchial biopsies. When switched at the air-liquid interface, the progenitor cells undergo a progressive differentiation and polarization to a fully ciliated epithelia. The mature MucilAir™ is composed of basal cells, ciliated cells and mucus cells. The proportion of these various cell types is preserved compared to what one observes in vivo (Huang et al., Drug Discovery and Development — Present and Future, 8, 2011, hereby incorporated by reference in its entirety). Epithelia were reconstituted with a mixture of cells isolated from 14 different normal nasal donors having ages and genders as specified in Table 1. Nasal epithelia were all fully differentiated at the start of the experiment. Table 1

[0085] MucilAir™ is functionally differentiated, secretes mucus and are electrically tight (TEER>200 Q.cm²). The activity of the main epithelial ionic channels, such as CFTR, EnaC, Na/K ATPase, is preserved and the epithelia is shown to respond in a regulated and vectorial manner to the pro-inflammatory stimulus, TNF-a. A large panel of cytokines, chemokines and metalloproteinases has been detected in MucilAir™ (e.g., IL-8, IL-6, GM-CSF, MMP-9, GRO- a). Most importantly, MucilAir™ replicates the main function of the airway epithelial cells, the mucociliary clearance driven by synchronized cilia-beating.

[0086] The SARS-CoV-2 strain used in the study was isolated by directly inoculating VeroE6 cell monolayers with a nasal swab sample collected from Bichat Claude Bernard Hospital, Paris. Once characteristic cytopathic effect was observable in more than 50% of the cell monolayer, supernatants were collected and immediately stored at -80°C. The complete viral genome sequence was obtained using Illumina MiSeq sequencing technology and was deposited under the name BetaCoV/France/IDF0571/2020. Viral stocks were titrated by tissue culture infectious dose 50 % (TCID50/mL) in VeroE6 cells, using Reed & Meunch statistical method.

[0087] Prior to infection, the apical side of the MucilAir™ cultures were washed twice for 10 min. Inoculations were performed with 150 pL at a theoretical multiplicity of infection (MOI) of 0.1 (50,000 TCID50 for an average of 500,000 cells in MucilAir™), applied to the apical side of the cultures for 1 hour at 37°C, 5% CO2. Non-infected control, Mock, was exposed also to 150 pL of culture medium on the apical side for 1 hour. Unbound viruses were removed after one hour of incubation period. New viral particles were collected by 10 minute apical washes 48 and 72 hours post inoculation (pi) and quantified by RT-qPCR.

[0088] Several stock solutions were prepared as shown in Table 2.

Table 2

f Stock 3 was prepared to form a 50:50 weight ratio mixture of lactoferrindysozyme using a 5 mg/mL lactoferrin in buffer (0.9% NaCl) solution and 100 mg/mL lysozyme in water solutions

[0089] Glycyrrhizin was obtained from Sigma and secretory leukocyte protease inhibitor (SLPI) was obtained from Origene Technologies. These four stock solutions were used to prepare 100 pL of the six test compositions shown in Table 3 using 0.9% NaCl buffer saline solution.

Table 3

[0090] As can be seen in Table 3, Test Compositions Al, A4, and A6 each comprised a concentration of 5 mg/mL glycyrrhizin and Test Compositions A2, A4, and A6 each comprised a concentration of 0.03 mg/mL SLPI. [0091] Each test composition was measured according to the protocol illustrated in FIGS. 1 A, IB, and 1C. These test compositions were compared against two negative vehicle controls (one with virus and one without) and a positive control comprising 5 pM of the antiviral Remdesivir which was applied basolaterally. Briefly, 10 pL of each test composition was applied to the apical portion of MucilAir™ media (t = -1 hr). One hour later, the apical side of the media having one of the test compositions dispersed thereon was inoculated with SARS- CoV-2 virions (French circulating strain) as described in detail above. The epithelia were inoculated with 100 pL inoculum (z.e., SARS-CoV-2; 500,000 virions, multiplicity of infection (MOI) = 0.1) for 1 hour at 37 °C. After 1 hour, the media were washed with PBS (with Ca²⁺/Mg²⁺ in order to remove the inoculum), using 150 pL of OptiMEM™ culture media. The media were then incubated in a 37 °C, 5% CO2, and 100% humidity environment for 1 hour. 10 pL of each test composition was reapplied to the apical surface at 1 hour post inoculation (t = 1 hr), 24 hours post inoculation (t = 24 hr), and 48 hours post inoculation (t = 48 hr) with incubation between each composition administration. For the positive control, 5 pM Remdesevir (purchased from MedChemExpress) and diluted in DMSO (DMSO final concentration was 0.05%) was added in the basolateral medium at 1, 24, and 48 hours post inoculation (pi).

[0092] Tissue integrity was also monitored using trans-epithelial electrical resistance (TEER) measurements at t = 48 hr and t = 72 hr. TEER is a dynamic parameter that reflects the state of epithelia and is typically from 200 to 600 Q.cm². An increase of the TEER value reflects a blockage of the ion channel activities. A notable decrease of the TEER values (but > 100 Q.cm²) could be observed in certain cases, reflecting an activation of ion channels. Disruption of cellular junction or holes in the epithelia result in TEER values below 100 Q.cm². After addition 200 pL of culture medium to the apical compartment of MucilAir™ cultures, resistance was measured with an EVOMX volt-ohm meter (World Precision Instruments UK, Stevanage) for each test condition. Resistance values (Q) were converted to TEER (Q.cm² using the following formula:

TEER (Q.cm²) = (resistance value (Q) - 100 (Q) * 0.33 (cm²) where 100 Q is the resistance of the membrane and 0.33 cm² is the total surface of the epithelium.

[0093] Apical washes of the epithelia were performed following the TEER measurements at 48 hours pi (t = 48 hr) and 72 hours pi (t = 72 hr) and prior to administration of any test composition with 200 pL of OptiMEM™ culture media during a 10 minute period with 37 °C environment. Samples collected from apical washes at different time-points were separated into 2 tubes: one for TCID50 viral titration and one for RT-qPCR. While RT-qPCR analysis was performed for all conditions, TCID50 was performed only for selected test conditions (Al, A4, and A6) chosen after RT-qPCR analysis.

[0094] From the 200 pL apical washes, 140 pL was used for viral RNA extraction with the QIAamp© Viral RNA kit (Qiagen), obtaining 60 pL of eluted RNA. Viral RNA was quantified by quantitative RT-PCR (EXPRESS One-Step Superscript™ qRT-PCR kit, Invitrogen , 1178101K) using 2 pL of viral RNA with Mastermix and two ORFlb-nspl4 specific primers (5’-TGGGGYTTTACRGGTAACCT-3’; 5’-AACRCGCTTAACAAAGCACTC-3’; 5’- AACRCGCTTAACAAAGCACTC-3’) and probe (5’-FAM-

TAGTTGTGATGCWATCATGACTAG-TAMRA-3’) of SARS-CoV-2 as designed by the School of Public Health/University of Hong Kong (Leo Poon, Daniel Chu, and Malik Peiris) and published by the World Health Organization as Assay 1 at https://www.who.int/docs/default-source/coronaviruse/peiris-protocol-16-l- 2O.pdf?sfvrsn=aflaac73_4, which is hereby incorporated by reference in its entirety. Samples were run on StepOnePlus™ Real-Time PCR System (Applied Biosystems). “W” as used in the probe sequence indicates a weakly interacting nucleotide at that position which may be A or T/U. Count (Ct) data were determined and relative changes in gene expression were calculated using the 2'^ACt method. Fold reductions for each measurement are shown in FIGS. 2A (t = 48 hr) and 2B (t = 72 hr) relative to the mean of the untreated infected inserts.

[0095] As can be seen in FIGS. 2A and 2B, a strong antiviral effect is seen with compositions comprising glycyrrhizin (Al, A4, and A6). Combination treatments of SLPI with glycyrrhizin (A4 and A6) also showed an antiviral effect. Antiviral control remdesiver reduced apical SARS-CoV-2 genome copies at both time points. The magnitude of inhibition was 3.4 log (mean Ct was 29) at 48 hours (vs. 0.9% NaCl with a mean Ct of 18.3) and 5.7 log (mean Ct was 31.9) at 72 hours (vs 0.9 % NaCl with a mean Ct of 13.2). The change at 72 hours was statistically significant from vehicle. Al, A4, and A6 administration strongly inhibited apical SARS-CoV-2 genome copies in MucilAir™ at both the 48 hour and 72 hour time points. Statistically significant (p<0.05) changes were observed with exposure to Al and A4 formulations at 48 hours and Al at 72 hours. Mean loglO reductions were 4.5, 4, and 3.5 for Al, A4, and A6, respectively, at 48 hours. Mean loglO reductions were 5.7, 5.6, and 5 for Al, A4, and A6, respectively at 72 hours. [0096] Mean Tissue Culture Infection Dose (TCID50) measurements were also performed on the apical washes TCID50 for samples Al, A4, and A6 confirming the efficacy of these test conditions. The appearance of clearly observable cytopathic effect of SARS-CoV-2 from 48 hours pi permitted the classic infection titre determination in cell culture, the Median Tissue Culture Infection Dose (TCID50), using African green monkey VeroE6 cell line. The assay enabled the validation of a large interval, range 1-8 loglO (TCID50) and showed high correlation (R² 0.94) with the molecular viral quantification RT-qPCR identified in Pizzorno A., et al. Cell Reports Medicine 1 (2020): 100059, which is hereby incorporated by reference in its entirety.

[0097] The results of the TCID50 measurements are shown in FIGS. 3A (48 hours pi) and 3B (72 hours pi). Reference Remdesivir showed a reduction of 1.5 and 3 loglO TCID50/mL in the number of SARS-CoV-2 infectious particles at 48 and 72 hours, respectively. All of Al, A4, and A6 strongly inhibited infectious SARS-CoV-2 number at both time points. Reduction of 3.1 and 2.6 loglO TCID50/mL were observed for Al at 48 and 72 hours, respectively, similar to Remdesivir. Exposure to A4 and A6 (glycyrrhizin in combination with SLPI) prevented all cytopathic effect of SARS-CoV-2 in VeroE6 cells. No dead cells were observed (detection limit of 63.2 TCID/mL). In test conditions involving administration of A4 and A6, the culture medium changed color and morphology of cells was different. Administration of glycyrrhizin and SLPI or fragments thereof, particularly in concentrations above 30 pg/mL, may result in increased activity on infected surfaces as compared to the use with either active alone. In comparing the rT-qPCT and TCID50 measurements, it can be seen that SLPI alone had minimal or no effect on SARS-CoV-2 infection, while a marked difference between is seen in the TCID50/mL measurements between administrations with glycyrrhizin alone in combination with SLPI.

[0098] The effect on tissue integrity of pre-treatment and repeated apical exposure to formulations is shown in FIG. 4. As discussed above, TEER was measured 2- and 3-days post infection (n=4, 3 cultures) and has a threshold limit value of lOOQ.cm². Non-infected cultures showed low TEER values (mean 204 Q.cm²) which are considered in the normal range of MucilAir™. TEER values may be interpreted as an “ON-OFF” parameter using the threshold limit of 100 Q.cm². In this “ON-OFF” type analysis, SARS-CoV-2 infection and exposure to A1-A6 did not change tissue integrity as TEER values remained above this threshold value at 48 and 72 hours. However, exposure to A4, A5, and A6 decreased TEER values at 72 hours upon SARS-CoV-2 infection, without disruption of tissue integrity. This decrease may suggest effects of the combination treatments on ion channel activities in the system.

[0099] As can be seen in FIG. 4, application of glycyrrhizin to the mucosa is shown to modulate TEER of the systems at both measured time points. When administered alone (Al), glycyrrhizin increased tissue integrity as measured by TEER as compared to positive control, and when administered with SLPI (A4 and A6), glycyrrhizin decreased TEER as compared to a positive control at measured both time points. Furthermore, SLPI alone (A2) had no effect on TEER. In either case (increased modulation or decreased modulation), the glycyrrhizin administration effected a change in the mucosal tissues that would not have been expected. For example, IntT Pub No WO 2019/046664, which is hereby incorporated by reference in its entirety, illustrates that glycyrrhizin alone had no antiviral effect or integrity effect on the infected tissues and at the amounts and regimens administered therein. In contrast, and without wishing to be bound by theory, the measured decrease in TEER correlating with substantial eradication of virions on infected tissues as demonstrated in FIGS. 2-4 (samples A4 and A6), may be due to the activation of separate heretofore unknown mechanisms afforded by administration of glycyrrhizin alone or in concert with another antiviral component. Highlighting this distinction, TEER decreased with sample A5 (composition without glycyrrhizin) as well, but the pronounced viral eradication measured was not present with this test condition. In both situations (increased or decreased modulation of TEER), administration of glycyrrhizin is a responsible factor for activity (e.g., anti-viral activity, changes to mucosal integrity which may stay above a threshold value such as 100 ) of the presently disclosed compositions.

[0100] Example 2

[0101] Several stock solutions were prepared as shown in Table 4.

Table 4

f Stock 2.3 was prepared to form a 50:50 weight ratio mixture of lactoferrin :lysozyme using a 5 mg/mL lactoferrin in buffer (PBS) from Ingredia (product name Proferrin) solution and 100 mg/mL lysozyme (Nutriscience) in water solutions [0102] Glycyrrhizin was obtained from Sigma and secretory leukocyte protease inhibitor (SLPI) was obtained from Origene Technologies. Following preparation, the vials were stored at -80°C until the first day of experiment. These four stock solutions were used to prepare 300 pL of the three test compositions shown in Table 5 using 0.9% NaCl buffer saline solution. Vials of each stock solution was thawed on ice prior to mixing.

Table 5

[0103] As can be seen, test compositions Bl, B2, and B3 had 5 mg/mL glycyrrhizin and test composition B3 had 0.1 mg/mL SLPI.

[0104] Test compositions were kept at 4° for application of Ih post-inoculation 7t, 4h pi, 24h pi, and 48h pi administration. Each test material was applied to a MucilAir media as described in Example 1 as shown in Table 6.

Table 6

[0105] TEER and RT-PCR were taken as described in Example 1. FIG. 5 shows the relative virus genome copy of Apical SARS-CoV-2 quantification at 48 hours and 72 hours pi for the three test conditions. Antiviral control Remdesivir reduced apical SARS-CoV-2 genome copies at both time points. The magnitude of inhibition was 3.1 loglO at 48 hours and 3.9 loglO at 72 hours (vs 0.9% NaCl). These changes were statistically significant from vehicle. Test compositions Bl and B2 inhibited apical SARS-CoV-2 genome copies (>1 loglO) in MucilAirt at both time points. The mean loglO reductions were 3 and 2.2 for Bl and B2 respectively, at 48 hours. The mean loglO reductions were 3.1 and 2.6 for Bl and B2, respectively, at 72 hours. Test composition B3, administered only following inoculation (z.e., in a treatment modality) showed minimal decrease at 48 hours pi, with an increase in viral reduction at 72 hours. This reduction was seen despite the higher order of magnitude concentrations of viral particles used in the assay as compared to what a person is exposed to, particularly in the early stages of infection.

[0106] Bl and B2 formulations were selected for TCID50 measurements at 48 and 72 hours. Those results are shown in FIGS. 6 A and 6B. Reference Remdesivir showed a reduction of 1.9 and 3 loglO TCID50/mL in the number of SARS-CoV-2 infectious particles at 48 and 72 hours respectively. Both selected formulations, Bl and B2 inhibited infectious SARS-CoV-2 number at both time points. The reductions were moderate at 48 hours, 0.8 and 0.9 loglO TCID50/mL for Bl and B2, respectively. The reductions were significant at 72 hours, 1.7 and 1.9 loglO TCID50/mL for Bl, and B2, respectively.

[0107] FIG. 7 provides the TEER measurements for each condition as well. In the Noninfected cultures (mock, 0.9% NaCl(-)), TEER values were measured in the normal range of MucilAir (200-600 Q.cm²). SARS-CoV-2 infection induced a slight decrease of TEER values at 72 hours, however TEERs stayed above the 100 Q.cm² thershold limit. Exposure to test compositions B 1 and B2, as well as Remdesivir, prevented virus induced TEER decrease, while exposure to B3 resulted in the same TEER value as the infected control NaCl(+).

[0108] Example 3

[0109] Experiments were performed on Human Coronavirus -229E as well. MucilAir as described in Example 1 was inoculated with 9.69* 10⁵ genome copies/mL (inoculation titer 1 x 10⁷ genome copies in 11 pL). The following treatment compositions were applied:

A: glycyrrhizin at a concentration of 5 mg/mL water;

B: dipotassium glycyrrhizate at a concentration of 5 mg/mL in 10 pL water

C SLPI at 18.75 pg/mL with glycyrrhizin at 5 mg/mL in 10 pL of water.

Dipotassium glycyrrhizinate hydrate (DPB) was ordered from Spectrum Chemical (TCI- G0270-25G), glycyrrhizin was orded from SigmaAldrich (item #CDS020796) and SLPL recomibinant human was ordered from Origen technologies (item #TP304697 Lot#WXOOACFF 0.05). [0110] The MucilAir cultures were placed in 24 well plates with 0.7 mL of MucilAir medium. 10 pL of treatment or buffer was added to the apical surface at incubated at 37°C for 30 minutes. 10 pL of virus was the applied apically. The plate was incubated at 33 °C for 3 hours. Tissues were washed 3 times with 200 pL of MucilAir medium. The plates were moved back to 33 °C. After 24 and 48 hours post inoculation, apical and basal medium were collected and frozen at -80°C. TEER was measured and the treatments were reapplied. After 72 hours, the apical and basal medium were collected and frozen at -80°C. The TEER was measured and the tissues were mixed in 4% formalin (PBS rinse, 4% PFA 10’ RT, PBS wash 3X, 4°C in PBS/axide).

[OHl] RNA was then extracted and reverse transcribed from the samples and analyzed by qPCR. RNA was extracted from 50 pL samples using the MagMax-96 viral isolation kit and following the user’s guide instructions. The final volume of RNA was 30pL (in elution buffer from the kit). Samples were stored at -20°C until use. cDNA synthesis was performed using the Superscript IV VILO kit. The annealing, reverse transcription, and enzyme inactivation steps were performed using the Quantstudio-3 machine in the lab. The final cDNA volume was 20 pL. qPCR was performed using the QuantiNova probe PCR kit using:

Probe: /56-FAM/TGTTGAAGA/ZEN/ACCTGACTCCCGTGC/3IABkFQ/

Primerl : AAGCTCCCAAATGGTGTTACT

Primer2: AGGATTCCGAGATTGAGGTTTG

Cycling conditions had an activation step of 2 minutes at 95°C with (Denaturation: 5sec at 95C Annealing/Extension: 30sec at 60C) x 40 cycles.

[0112] FIGS. 8 A and 8B provide the measured genome copies/mL from each sample at 24 hours (FIG. 8A) and 48 hours (FIG. 8B). Error bars represent the standard deviation of the measurements illustrating that application of compositions A, B, and C resulting in statistically significant viral reductions. Furthermore, the glycyrrhizate salt (dipotassium glycyrrhizate, composition B) showed increased reduction in genome copy number than glycyrrhizin (composition C) at 48 hours pi. The combination of glycyrrhizin and SLPI showed the greatest inhibition of viral spread.

[0113] As various changes can be made in the above-described subject matter without departing from the scope and spirit of the present disclosure, it is intended that all subject matter contained in the above description, or defined in the appended claims, be interpreted as descriptive and illustrative of the present disclosure. Many modifications and variations of the present disclosure are possible in light of the above teachings. Accordingly, the present description is intended to embrace all such alternatives, modifications and variances which fall within the scope of the appended claims.

[0114] All documents cited or referenced herein and all documents cited or referenced in the herein cited documents, together with any manufacturer's instructions, descriptions, product specifications, and product sheets for any products mentioned herein or in any document incorporated by reference herein, are hereby incorporated by reference, and may be employed in the practice of the disclosure.

Claims

1. A method for the treatment of viral infection (e.g., respiratory viral infection, influenza, coronavirus such as SARS-CoV-2, Coronavirus 229E) comprising topically administering a pharmaceutical composition to a subject in need thereof (e.g., a subject having a viral load of said SARS-CoV-2), wherein said pharmaceutical composition comprises one or more pharmaceutically acceptable excipients and glycyrrhizin or, enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium, dipotassium glycyrrhizinate) and/or prodrugs or hydrates of any of the foregoing.

2. The method according to claim 1, wherein said topical administration is topical administration to the oral or nasal mucosa.

3. The method according to claim 1 or 2, wherein said pharmaceutical composition is an oral spray, a nasal spray, or a lozenge.

4. The method according to any one of claims 1-3, wherein said pharmaceutical composition comprises less than (or from 0.1% to) 50% (e.g., less than 40%, less than 30%, less than 20%, less than 10%, from 0.001 to 0.01%, from 0.01% to 0.1%, from 0.1% to 1%, from 1% to 10%, from 10% to 50%) glycyrrhizin by weight of the composition.

5. The method according to any one of claims 1-4, wherein said pharmaceutical composition comprises a concentration of glycyrrhizin of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 10 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 5 mg/mL).

6. The method according to any one of claims 1-5, wherein said pharmaceutical composition is formulated to provide a concentration of glycyrrhizin on the area of topical administration (e.g., the oral mucosa, the nasal mucosa) of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL, from 1 mg/mL to 50 mg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 25 mg/mL).

7. The method according to any one of claims 1-6, wherein said glycyrrhizin is a pharmaceutically acceptable salt of glycyrrhizin.

8. The method according to any one of claims 1-6, wherein said glycyrrhizin is dipotassium glycyrrhizinate.

9. The method according to any one of claims 1-8, wherein said infection is an infection of SARS-CoV-2 and viral load of said SARS-CoV-2 in said subject at 48 hours following administration is less than (or from 0.0001% to) 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of said compound) .

10. The method according to any one of claims 1-9, wherein said infection is an infection of SARS-CoV-2 and wherein viral load of said SARS-CoV-2 in said subject at 72 hours is less than (or from 0.0001% to) 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of said compound).

11. The method according to any one of claims 1-10, wherein said administration is daily administration.

12. The method according to any one of claims 1-11, wherein said glycyrrhizin is the sole active ingredient in the pharmaceutical composition.

13. The method according to any one of claims 1-11, wherein said pharmaceutical composition comprises an additional antiviral agent.

14. The method according to claim 13, wherein said additional antiviral agent is an enzyme (e.g., a protease inhibitor such as secretory leukocyte protease inhibitor (SLPI)).

15. The method according to claim 13, wherein the ratio of said glychyrrhizimadditional antiviral agent (e.g., SLPI) is 100:1 to 1:100 (e.g., 50:1 to 1:50, 25:1 to 1:25, 20:1 to 1:20, 10:1 to 1:10, 5:1 to 1:5, 2:1 to 1:2, 1:1 to 1:100, 1:1 to 1:50, 1:1 to 1:25, 1:1 to 1:20, 1:1 to 1:10, 1:1 to 1:5, 1:1 to 1:2, 100:1 to 1:1, 50:1 to 1:1, 25:1 to 1:1, 20:1 to 1:1, 10:1 to 1:1, 5:1 to 1:1, 2:1 to 1:1, 20:1 to 10:1, 20:1 to 15:1, 17:1 to 16:1).

16. The method according to any one of claims 1-14, wherein said pharmaceutical composition further comprises a glycoprotein (e.g, sialic acid such as siallyllactose, 6’- sialyllactose, 3’-sialyllactose combinations of 6’-sialyllactose and 3-sialyllactose such as in a 10: 1 to 1 : 10 or 5: 1 to 1 :5 or 2: 1 to 1 :2 weight ratio).

17. The method according to any one of claims 1-16, wherein said pharmaceutical composition comprises lactoferrin (e.g., human lactoferrin, apolactoferrin, bovine lactoferrin) and/or lysozyme (e.g., chicken lysozyme).

18. The method according to any one of claims 1-17, wherein the viral RNA of said SARS-CoV-2 comprise the sequence ORFlb-nspl4 or fragments thereof (e.g., ORFlb- nspl4F, ORFlb-nspl4R, ORFlb-nspl4P).

19. The method according to claim 18, wherein the viral RNA of said SARS-CoV-2 comprise one or more sequences complimentary to:

5 ’ -TGGGGYTTTACRGGTAACCT-3 ’ ,

5’-AACRCGCTTAACAAAGCACTC-3’, or 5’-TAGTTGTGATGCWATCATGACTAG- 3’.

20. The method according to any one of claims 1-19, wherein the administration delivers said glycyrrhizin to the oral and/or nasal mucosa (e.g., coats the oral and/or nasal mucosa or a portion thereof with a composition comprising glycyrrhizin) and the viral load of said virions (e.g., SARS-CoV-2) on the oral/and nasal mucosa at 48 hours following administration is less than 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of said compound).

21. The method according to any one of claims 1-20, wherein the administration delivers said glycyrrhizin to the oral and/or nasal mucosa (e.g., coats the oral and/or nasal mucosa or a portion thereof with a composition comprising glycyrrhizin) and the viral load (e.g., SARS- CoV-2) on the oral/and nasal mucosa at 72 hours following administration is less than 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of said compound).

22. The method according to any one of claims 1-21, wherein said administration delivers said glycyrrhizin to the oral and/or nasal mucosa of said subject and said composition modulates the tissue integrity of said oral and/or nasal mucosa (e.g., as measured by TEER).

23. The method according to any one of claims 1-21, wherein said administration delivers said glycyrrhizin to the oral and/or nasal mucosa of said subject and said composition increases the tissue integrity of said oral and/or nasal mucosa (e.g., as measured by TEER) as compared to the tissue integrity of otherwise identical mucosa not having said composition administered thereto.

24. The method according to any one of claims 1-21, wherein said administration delivers said glycyrrhizin to the oral and/or nasal mucosa of said subject and said composition increases the tissue integrity of said oral and/or nasal mucosa (e.g., as measured by TEER at 48 hours following infection) as compared to the tissue integrity of otherwise identical mucosa not having said composition administered thereto.

25. The method according to any one of claims 1-21, wherein said administration delivers said glycyrrhizin to the oral and/or nasal mucosa of said subject and said composition increases the tissue integrity of said oral and/or nasal mucosa (e.g., as measured by TEER at 72 hours following infection) as compared to the tissue integrity of otherwise identical mucosa not having said composition administered thereto.

26. The method according to any one of claims 1-21, wherein said administration delivers said glycyrrhizin to the oral and/or nasal mucosa of said subject and said composition decreases the tissue integrity of said oral and/or nasal mucosa (e.g., as measured by TEER at 48 hours following infection) as compared to the tissue integrity of otherwise identical mucosa not having said composition administered thereto.

27. The method according to any one of claims 1-21, wherein said administration delivers said glycyrrhizin to the oral and/or nasal mucosa of said subject and said composition decreases the tissue integrity of said oral and/or nasal mucosa (e.g., as measured by TEER at 72 hours following infection) as compared to the tissue integrity of otherwise identical mucosa not having said composition administered thereto.

28. The method according to any one of claims 1-27, wherein said viral infection is SARS-CoV-2 infection.

29. The method according to any one of claims 1-27, wherein said viral infection is Coronavirus -229E infection.

30. A method for the prophylaxis of viral infection (e.g., coronavirus such as SARS-CoV- 2 infection) comprising topical administration of a pharmaceutical composition to a subject in need thereof (e.g., a subject having a viral load of said SARS-CoV-2), wherein said pharmaceutical composition comprises one or more pharmaceutically acceptable excipients and glycyrrhizin or, enantiomers thereof, racemic mixtures thereof, mixtures of enantiomers thereof, or pharmaceutically acceptable salts (e.g., glycyrrhizin ammonium, glycyrrhizin diammonium) and/or prodrugs of any of the foregoing.

31. The method according to claim 30, wherein said topical administration is topical administration to the oral or nasal mucosa.

32. The method according to claim 30 or 31, wherein said pharmaceutical composition is an oral spray, a nasal spray, or a lozenge.

33. The method according to any one of claims 30-32, wherein said pharmaceutical composition comprises less than (or from 0.1% to) 50% (e.g., less than 40%, less than 30%, less than 20%, less than 10%, from 0.001 to 0.01%, from 0.01% to 0.1%, from 0.1% to 1%, from 1% to 10%, from 10% to 50%) glycyrrhizin by weight of the composition.

34. The method according to any one of claims 30-33, wherein said pharmaceutical composition comprises a concentration of glycyrrhizin of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 10 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 5 mg/mL).

35. The method according to any one of claims 30-34, wherein said glycyrrhizin is a pharmaceutically acceptable salt of glycyrrhizin.

36. The method according to any one of claims 30-34, wherein said glycyrrhizin is dipotassium glycyrrhizinate.

37. The method according to any one of claims 30-36, wherein said pharmaceutical composition is formulated to provide a concentration of glycyrrhizin on the area of topical administration (e.g., the oral mucosa, the nasal mucosa) of less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL, from 1 mg/mL to 50 mg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 25 mg/mL).

38. The method according to any one of claims 30-37, wherein said infection is an infection of SARS-CoV-2 and wherein viral load of said SARS-CoV-2 in said subject at 48 hours following administration is less than 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of said compound).

39. The method according to any one of claims 30-38, wherein said infection is an infection of SARS-CoV-2 and wherein viral load of said SARS-CoV-2 in said subject at 72 hours is less than (or from 0.01% to) 10% (e.g., less than 1%, less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of said compound).

40. The method according to any one of claims 30-37, wherein said infection is an infection of SARS-CoV-2 and wherein the administration delivers said glycyrrhizin to the oral and/or nasal mucosa (e.g., coats the oral and/or nasal mucosa or a portion thereof with a composition comprising glycyrrhizin) and the viral load of said SARS-CoV-2 on the oral/and nasal mucosa at 48 hours following administration is less than 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of said compound).

41. The method according to any one of claims 30-37 or 40, wherein the administration delivers said glycyrrhizin to the oral and/or nasal mucosa (e.g., coats the oral and/or nasal mucosa or a portion thereof with a composition comprising glycyrrhizin) and the viral load of said SARS-CoV-2 on the oral/and nasal mucosa at 72 hours following administration is less than 1% (e.g., less than 0.1%, less than 0.05%) as compared to the viral load prior to administration (e.g., the viral load less than 1 day or less than 12 hours or less than 6 hours, or less than 3 hours or less than 1 hour prior to the first administration of said compound).

42. The method according to any one of claims 30-41, wherein said administration is daily administration.

43. The method according to any one of claims 30-41, wherein said administration occurs two or more (e.g., two, three, four) times per day.

44. The method according to any one of claims 30-42, wherein said glycyrrhizin is the sole active ingredient in the pharmaceutical composition.

45. The method according to any one of claims 30-42, wherein said pharmaceutical composition comprises an additional antiviral agent.

46. The method according to claim 45, wherein said additional antiviral agent is an enzyme (e.g., protease inhibitor, secretory leukocyte protease inhibitor (SLPI)).

47. The method according to claim 45, wherein the ratio of said glychyrrhizimadditional antiviral agent (e.g., protease inhibitor such as SLPI) is 100:1 to 1:100 (e.g., 50:1 to 1:50, 25:1 to 1:25, 20:1 to 1:20, 10:1 to 1:10, 5:1 to 1:5, 2:1 to 1:2, 1:1 to 1:100, 1:1 to 1:50, 1:1 to 1:25, 1:1 to 1:20, 1:1 to 1:10, 1:1 to 1:5, 1:1 to 1:2, 100:1 to 1:1, 50:1 to 1:1, 25:1 to 1:1, 20:1 to 1:1, 10:1 to 1:1, 5:1 to 1:1, 2:1 to 1:1, 20:1 to 10:1, 20:1 to 15:1, 17:1 to 16:1).

48. The method according to any one of claims 30-46, wherein said pharmaceutical composition further comprises a glycoprotein (e.g., sialic acid such as siallyllactose, 6’- sialyllactose, 3’-sialyllactose combinations of 6’-sialyllactose and 3-sialyllactose such as in a 10:1 to 1:10 or 5:1 to 1:5 or 2:1 to 1:2 weight ratio).

49. The method according to any one of claims 30-48, wherein said pharmaceutical composition comprises lactoferrin (e.g., human lactoferrin, apolactoferrin) and/or lysozyme (e.g., chicken lysozyme).

50. The method according to any one of claims 30-49, wherein the viral RNA of said SARS-CoV-2 comprise the sequence ORFlb-nspl4 or fragments thereof (e.g., ORFlb- nspl4F, ORFlb-nspl4R, ORFlb-nspl4P).

51. The method according to claim 50, wherein the viral RNA of said SARS-CoV-2 comprise one or more sequences complimentary to:

5 ’ -TGGGGYTTTACRGGTAACCT-3 ’ ,

5’-AACRCGCTTAACAAAGCACTC-3’, or

5’-TAGTTGTGATGCWATCATGACTAG- 3’.

52. The method according to any one of claims 30-51, wherein said viral infection is SARS-CoV-2 infection.

53. The method according to any one of claims 30-51, wherein said viral infection is Coronavirus -229E infection.

54. A method for the treatment and/or prophylaxis of respiratory infection comprising administering a pharmaceutical composition comprising glycyrrhizin and a pharmaceutically acceptable carrier, excipient, and diluent to the oral and/or nasal mucosa of a subject in need thereof; wherein said pharmaceutical composition comprises less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 10 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 5 mg/mL) glycyrrhizin; wherein said glycyrrhizin is administered in an amount sufficient to reduce the viral infection in said subject (e.g., at 48 hours following infection or exposure to infection, at 72 hours following infection or exposure to infection).

55. The method according to claim 54, wherein said respiratory infection is SARS-CoV-2 infection.

56. The method according to claim 54, wherein said respiratory infection is Coronavirus - 229E infection.

57. A method comprising administering a pharmaceutical composition comprising glycyrrhizin and a pharmaceutically acceptable carrier, excipient, and diluent to a subject having a viral infection or a subject having been exposed to a viral infection; wherein said pharmaceutical composition comprises less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 5 mg/mL) glycyrrhizin; and wherein said glycyrrhizin is administered in an amount sufficient to reduce the viral infection in said subject (e.g., at 48 hours following infection or exposure to infection, at 72 hours following infection or exposure to infection).

58. The method according to claim 57, wherein said respiratory infection is SARS-CoV-2 infection.

59. The method according to claim 57, wherein said respiratory infection is Coronavirus - 229E infection.

60. A method comprising administering a pharmaceutical composition comprising glycyrrhizin and a pharmaceutically acceptable carrier, excipient, and diluent to the oral and/or nasal mucosa of a subject having a viral infection or a subject having been exposed to a viral infection; wherein said pharmaceutical composition comprises less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 10 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 5 mg/mL) glycyrrhizin; and wherein said glycyrrhizin is administered in an amount sufficient to modulate (e.g., increase, decrease) the tissue integrity of the oral and/or nasal mucosa (e.g., as compared to the tissue of otherwise identical mucosa at, for instance, 48 hours or 72 hours, following initial administration of said composition).

61. The method according to claim 60, wherein said tissue has been infected a virus (e.g., coronavirus, SARS-CoV-2, Coronavirus -229E).

62. The method according to any one of claims 54-61, wherein said glycyrrhizin is a pharmaceutically acceptable salt of glycyrrhizin.

63. The method according to any one of claims 54-61, wherein said glycyrrhizin is dipotassium glycyrrhizinate.

64. A pharmaceutical composition formulated for delivery to the oral and/or nasal mucosa, wherein said pharmaceutical composition comprises a pharmaceutically acceptable carrier, excipient, and diluent; and less than (or from 0.1 mg/mL to) 2400 mg/mL (e.g., less than 2000 mg/mL, less than 1000 mg/mL, less than 600 mg/mL, less than 300 mg/mL, less than 100 mg/mL, less than 10 mg/mL, less than 5 mg/mL, less than 1000 pg/mL, less than 500 pg/mL, less than 100 pg/mL, less than 50 pg/mL, less than 30 pg/mL, from 1 pg/mL to 100 pg/mL, from 100 pg/mL to 1000 pg/mL, from 1 mg/mL to 10 mg/mL, from 1 mg/mL to 5 mg/mL) glycyrrhizin; wherein said composition is characterized as resulting in a modulation of the tissue integrity of said oral and/or nasal mucosa following application thereto.

65. The pharmaceutical composition according to claim 64, wherein said oral and/or nasal mucosa are infected with virions (e.g., influenza virions, coronavirus virions such as SARS- CoV-2).

66. The pharmaceutical composition according to claim 65, wherein said pharmaceutical composition is further characterized as reducing the number of virions following application of the composition to said mucosa (e.g., as compared to administration of an otherwise identical composition not comprising said glycyrrhizin, for example at from 24 hours to 72 hours following administration).

67. The pharmaceutical composition according to any one of claims 64-66, wherein said glycyrrhizin is a pharmaceutically acceptable salt of glycyrrhizin.

68. The method according to any one of claims 64-66, wherein said glycyrrhizin is dipotassium glycyrrhizinate.

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