NOVEL PREVENTIVE AND THERAPEUTIC TREATMENT FOR COVID 19 AND ANY OTHER DISEASE CAUSED BY SARS COV 2

Abstract

Novel preventive treatment for COVID 19 and therapeutic treatment for early stages of COVID 19 or any other disease caused by SARS CoV 2. Pharmaceutical formulations and devices are disclosed to be able to implement these treatments. These treatments to prevent and alleviate the symptoms at early stages of COVID 19, the pandemic disease caused by SARS CoV 2. These treatments are based on the administration the nasal cavity and/or the lungs of solutions containing iota Carrageenan with or without the addition of xylitol as antiviral drug substances.

Claims

1. A nasal spray comprising a solution having about 0.001% w/v to about 0.50% w/v iota carrageenan and about 0.5% w/v to about 10% w/v of xylitol as active agents.

2. The nasal spray of claim 1, wherein the solution comprises one or more preservatives comprising potassium sorbate, sorbic acid, sodium benzoate, benzoic acid, methylparaben, propylparaben, benzyl alcohol, phenoxyethanol or combinations thereof.

3. The nasal spray claim 1, wherein the nasal spray is administered via a pump dispensing 10 to 200 μL of solution per puff.

4. The nasal spray of claim 1, wherein the solution further comprises one or more of: osmolytes, buffers and combinations thereof.

5. The nasal spray of claim 1, wherein the osmolytes comprise at least one of: sodium chloride, mannitol, sorbitol or combinations thereof.

6. The nasal spray of claim 1, wherein the buffers comprise at least one of: citric acid/sodium citrate, disodium hydrogen phosphate/sodium dihydrogen phosphate, acetic acid/sodium acetate or combinations thereof.

7. The nasal spray of claim 1, wherein the active agents are anti-viral agents.

8. The nasal spray of claim 7, wherein the anti-viral agents prevent or inhibit coronavirus infections.

9. The nasal spray of claim 8, wherein the coronavirus is SARS CoV 2.

10. A method of treating a coronavirus infection comprising: administering a nasal spray comprising a solution having about 0.001% w/v to about 0.50% w/v iota carrageenan and about 0.5% w/v to about 10% w/v of xylitol as active agents, wherein the solution comprises one or more preservatives comprising potassium sorbate, sorbic acid, sodium benzoate, benzoic acid, methylparaben, propylparaben, benzyl alcohol, phenoxyethanol or combinations thereof, thereby, reducing the viral load in the nose and treating the coronavirus infection.

11. The method of claim 10, wherein the nasal spray is administered via a pump dispensing 10 to 200 μL of solution per puff.

12. The method of claim 10, wherein the solution further comprises one or more of: osmolytes, buffers and combinations thereof.

13. The method of claim 10, wherein the active agents prevent or inhibit respiratory viral infections.

14. The method of claim 13, wherein the virus is SARS CoV 2.

15. A nasal spray comprising a solution having about 0.001% w/v to about 0.50% w/v iota carrageenan and about 0.3% w/v to about 0.9% w/v of sodium chloride as osmolyte.

16. The nasal spray of claim 15, wherein the solution further comprises preservatives from the group benzalkonium chloride, sorbic acid, potassium sorbate, benzyl alcohol, phenoxyethanol, disodium edetate, edetic acid, methyl paraben, propyl paraben and/or one or more osmolytes selected from the group consisting of mannitol, sorbitol, and glucose.

17. The nasal spray of claim 15, wherein the iota-carrageenan is an anti-viral agent.

18. The nasal spray of claim 15, wherein iota/carrageenan prevent or inhibit respiratory viral infections.

19. The nasal spray of claim 18, wherein the coronavirus is SARS CoV 2.

20. The nasal spray of claim 15, wherein the solution comprises iota carrageenan 0.17% w/v, sodium chloride 0.9% w/v, sodium hydroxide or hydrochloric acid to adjust pH to 6-7 in a nasal spray with a 100 microliter pump.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0054] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawings will be provided by the Office upon request and payment of the necessary fee.

[0055] FIG. 1 is a graph demonstrating the effect of iota carrageenan on SARS-CoV-2 with Formulation #1 (1.7 mg/mL IC, 0.9% sodium chloride, 0% xylitol, pH 6-7).

[0056] FIG. 2 is a graph demonstrating the effect of iota carrageenan on SARS-CoV-2 Formulation #2 (1.2 mg/mL IC, 0.5% sodium chloride, 0% xylitol, pH 6-7).

[0057] FIG. 3 is a graph demonstrating the effect of iota carrageenan on SARS-CoV-2 Formulation #3 (1.2 mg/mL IC, 0% sodium chloride, 0.5% xylitol, pH 6-7).

[0058] In FIGS. 1-3, Vero E6 cells were treated with 600 μg/ml, 60 μg/ml, 6 μg/ml or 0.6 μg/ml pre- and post-infection with SARS-CoV-2. After a 2 h pretreatment, cells were infected with SARS-CoV-2 and incubated for 48 h in the presence of IC. Supernatants were harvested and virus yield determined by an end point dilution assay (TCID50). Controls consisted of untreated infected cells or infected cells treated with formulation without IC. Results were determined using the Reed and Muench formula and expressed as TCID50/ml. Dotted line shows the limit of detection (LOD). Testing of samples was performed in triplicate.

DETAILED DESCRIPTION

[0059] The present disclosure is based, in part, on the unexpected and surprising discovery that iota Carrageenan at a concentration of not less than 6 μg/mL, a combination of iota Carrageenan and xylitol with a concentration of not less than 0.6 μg/mL of iota Carrageenan and xylitol alone are all capable of inhibiting SARS CoV 2 in vitro. These surprising results are summarized in Example 1.

[0060] Carrageenans are linear sulfated polysaccharides that are often extracted from red seaweeds. Carrageenans are commercially available in the form of kappa (κ), iota (.Math.) or lambda (λ). Clinical trials with a formulation containing xylometazoline and iota Carrageenan revealed that the latter stays in the nasal mucosa for about 4 hrs. ((Graf et al., 2018, International Journal of General Medicine 2018:11 275-283), which makes it a good candidate for an antiviral nasal spray, if proved to be effective either in preventing virus cell infection and/or lowering viral load for the particular pathogen in question.

[0061] Xylitol is a polyol used as sugar substitute for diabetic patients. It is safe and have shown some antiviral properties against Influenza virus and human respiratory syncytial virus when ingested (Xu et al., Biol. Pharm. Bull. 39, 540-546 (2016); Yin S. Y. et al. (2014) PLoS ONE 9(1): e84633. doi: 10.1371/journal.pone.0084633). Xylitol is used as an osmolyte in nasal sprays, as well.

[0062] Both iota carrageenan and xylitol are safe for humans, being used in much larger amounts as food additive and sweetener, respectively, than those that may be used for nasal delivery. Nasal safety of iota Carrageenan by nasal and nebulization administration has been already confirmed empirically (Hebar A. et al. (2015), PLoS ONE 10(4): e0122911. doi: 10.1371/journal.pone.0122911). The same holds for 5% Xylitol water solution both applied as nasal spray and nasal irrigation (Weissman J. D. et al., Laryngoscope, 121:2468-2472, 2011), as well as applied as a nebulization solution (Durairaj L. et al., 2004, Respiratory Research 2004, 5:13 doi:10.1186/1465-9921-5-13).

[0063] In another embodiment, a solution comprises: iota carrageenan from about 0.01 to about 0.5% w/v, iota Carrageenan from about 0.001 to 0.5% w/v and xylitol in a 0.5-10% w/v or xylitol in a concentration from 0.5 to 10% w/v. In certain embodiments, the solution comprises an osmolyte, for example, sodium chloride, mannitol, sorbitol, glycerin or any other well known in the art. The said formulation is packaged into a nasal spray to be delivered to the nose as one puff of 50-200 μL into each nostril each 4 hours during waking hours. In certain embodiments, The solution may contain suitable buffers, such citric acid/sodium citrate, disodium phosphate, dihydrogen sodium phosphate, acetic acid/sodium acetate or other buffers known in the art. This formulation is based on the surprising result of Example 1 and the following estimations: iota carrageenan has a residence time of approximately 4 hour; taking into account that surface area of the nasal mucosa is 100-250 cm.sup.2 (Bitter C et al. (2011), Curr Probl Dermatol. Basel, Karger, 2011, vol 40, pp 20-35, Garcia G J M et al. (2008), Inhalation Toxicology, 2009; 21(7): 607-618, DOI: 10.1080/08958370802320186, Gizurarzon S (2012), Current Drug Delivery, 2012, 9, 566-582, Pires A et al. (2009), J Pharm Pharmaceut Sci 12(3) 288-311, 2009) and that the airway surface liquid height, though variable, would be 5-15 μm (Helassa N et al. (2014), Biochem. J., (2014) 464, 213-220, doi:10.1042/BJ20141041, Wagenman et al. (1992), J Allergy Clin Immunol, Vol. 90, No. 3, Part 2, pp 419-423), the airway surface liquid volume is within 50 and 375 μL.

[0064] Accordingly, administering a 0.01% w/v of iota carrageenan and 5% xylitol in 1 puff of 100 μl per nostril, provides a nasal concentration of iota carrageenan of at least 35 μg/mL and 1.74% w/v xylitol used as an osmolyte enhances further the antiviral action in vivo by lowering the ionic strength of the nasal airway surface liquid and thus improving the innate antiviral action of defensins (Klotman M E et al., 2006, Nature Reviews Immunology, Vol. 6, June 2006, pp 447-456, Furci L et al. 2012, PLoS ONE 7(9): e45208. doi: 10.1371/journal.pone.0045208).

[0065] The nasal formulation may be sterilized by filtration as taught by (Grassauer 2016, CA 2992352 A1) and filled aseptically into suitable bottles, snapping appropriate preservative free nasal pumps onto them. Possible realizations of this embodiment are described in Examples 2, 3, 4 and 5.

[0066] In another embodiment, a solution comprises: iota carrageenan from about 0.01 to about 0.5% w/v, iota Carrageenan from about 0.001 to 0.5% w/v and xylitol in a 0.5-10% w/v or xylitol in a concentration from 0.5 to 10% w/v. An osmolyte may be added from the group: sodium chloride, mannitol, sorbitol, glycerin or any other well known in the art. This solution also contains sorbic acid, potassium sorbate, parabens, benzalkonium chloride, benzoic acid, sodium benzoate, phenoxyethanol, phenyl ethanol or other suitable preservatives known in the art with or without the addition of chelating agents, such as citric acids, sodium citrate or sodium salts of edetic acid may be added to the formulation to preserve it, allowing a non-sterile nasal formulation in bottles fitted with standard nasal pumps as taught in Examples 6, 7 and 8.

[0067] In another embodiment a solution for either nasal or lung nebulization solution comprises: iota carrageenan from about 0.01 to about 0.5% w/v, iota Carrageenan from about 0.001 to 0.5% w/v and xylitol in a 0.5-10% w/v or xylitol in a concentration from 0.5 to 10% w/v. In certain embodiments, the solution further comprises sodium chloride or another suitable osmolyte such as mannitol, sorbitol, and others well known in the art is packaged into unpreserved unit dose vials to be used for the nebulization of patients. The solution may contain suitable buffers, such citric acid/sodium citrate, disodium phosphate, dihydrogen sodium phosphate or other buffers known in the art. In this pharmaceutical form higher volumes are usually employed and, therefore, even lower concentrations of iota carrageenan allow the administration of suitable mass doses of this drug substance. At the same time, nasal nebulization reaches a more extensive area of the nasal cavity, particularly the deeper and higher portions (Moffa 2019). The nasal nebulization is particularly useful in treating patients in early stages of a respiratory disease caused by SARS CoV 2, such as pandemic COVID 19. Lung nebulization is helpful for patients and may be practiced by using the same composition in a nebulizer. This embodiment is illustrated in examples 9, 10 and 11.

[0068] In still another embodiment, a solution for either nasal or lung nebulization solution comprises: iota carrageenan from about 0.01 to about 0.5% w/v, iota Carrageenan from about 0.001 to 0.5% w/v and xylitol in a 0.5-10% w/v or xylitol in a concentration from 0.5 to 10% w/v with the possible addition of sodium chloride or another suitable osmolyte such as mannitol, sorbitol, and others well known in the art. This solution also contains sorbic acid, potassium sorbate, parabens, benzalkonium chloride, benzoic acid, sodium benzoate, phenoxyethanol, phenyl ethanol or other suitable preservatives known in the art with or without the addition of chelating agents, such as citric acids, sodium citrate or sodium salts of edetic acid may be added to the formulation to preserve it. This solution is packaged into multidose vials to be used for the nasal and/or lung nebulization of patients at early stages of the disease. This embodiment is illustrated in examples 12 and 13.

[0069] In another embodiment, a solution for nasal lavage or nasal wash comprises: iota carrageenan from about 0.01 to about 0.5% w/v, iota Carrageenan from about 0.001 to 0.5% w/v and xylitol in a 0.5-10% w/v or xylitol in a concentration from 0.5 to 10% w/v. with the possible addition of sodium chloride or another suitable osmolyte such as mannitol, sorbitol, and others well known in the art is packaged into either spray bottles or aerosol cans fitted with continuous valves. The formulation may contain sorbic acid, potassium sorbate, parabens, benzalkonium chloride, benzoic acid, sodium benzoate, phenoxyethanol, phenyl ethanol or other suitable preservatives known in the art with or without the addition of chelating agents, such as citric acids, sodium citrate or sodium salts of edetic acid, allowing for a preserved non-sterile nasal wash formulation. This practical device helps rinse the nose with these active ingredients and may eliminate more thoroughly viruses present in the whole of the nasal cavity. They are particularly suitable for the early stages of SARS CoV 2 infection. This embodiment is illustrated in examples 14 and 15.

[0070] Accordingly, the compositions referred to herein comprising iota and/or kappa carrageenan is also active against coronaviruses, e.g. COVID-19, including other respiratory viruses such as, human rhinovirus, a member of the paramyxoviridae such as parainfluenza virus, metapneumovirus, or respiratory syncytial virus, a member of the orthomyxoviridae such as influenza virus, or an adenovirus subtype B(ICD-10 codes J09 and J10).

[0071] The composition can include a pharmaceutically acceptable carrier, e.g., one or more solvents, dispersion media, coatings, antimicrobial agents, isotonic and absorption delaying agents, and the like, compatible with administration to a mammal, such as a human. Any carrier compatible with the excipient(s) and therapeutic agent(s) is suitable for use. Supplementary active compounds may also be incorporated into the compositions.

[0072] In certain embodiments, the pharmaceutical compositions are specifically adapted for topical administration to the nasal cavity. The pharmaceutical compositions may be applied before or after the outbreak of a respiratory viral infection in a human individual. Even if applied after the outbreak of a viral infection the compositions still prevent or at least ameliorate late complications of respiratory viral infections. Such complications are known in the art and include—but are not limited to—complications in connection with secondary infections by bacteria, and deterioration of pre-existing diseases such as allergy or COPD.

[0073] Topically administrable intranasal compositions referred to herein may have a pH value within a range of from 3.5 to 8.0, usually within a range of from about 4.0 to about 8.0. They may comprise one or more nasally compatible pH adjusting agents or buffer systems that prevent pH drift during storage. Such pH adjusting agents include, but are not limited to, boric acid, sodium borate, potassium citrate, citric acid, sodium bicarbonate, and various inorganic phosphate buffers such as Na.sub.2HPO.sub.4, NaH.sub.2PO.sub.4, KH.sub.2PO.sub.4, and mixtures thereof. The minimal ionic strengths introduced by any such pH-adjusting agents do not interfere with the essence of the invention. To prevent precipitation of calcium with phosphate ions from the buffer system, EDTA may be added up to a concentration of 2 mg/ml. In addition, flavors such as eucalyptus, campher, menthol, peppermint or similar, by way of oils or extracts, may be added to the product at concentrations known in the art.

[0074] Also, the topical intranasal formulations referred to herein may comprise one or more intranasally compatible surfactants. The surfactant facilitates the spread of the formulation across the surface of the nasal mucosa and may be non-ionic or anionic. Exemplary non-ionic surfactants may be selected from the group comprising tyloxapol, polyoxyethylene sorbitan esters, polyethoxylated castor oils, poloxamers, polyoxyethylene/polyoxypropylene surfactants, polyoxyethylene stearate, polyoxyethylene propylene glycol stearate, hydroxyalkylphosphonate, lauric or palmitic acid esters and ethers, triethanol amine oleate, or from a combination of the foregoing agents. Still further suitable surfactants may be known to those skilled in the art. The surfactants may typically be present at concentrations of from 0.02% (w/v) to 0.1% (w/v) of the composition.

[0075] In various embodiments, the present topical intranasal preparation may contain one or more preservatives to inhibit microbial growth and to prolong shelf life. Exemplary preservatives include, but are not limited to, disodium edetate (EDTA) and potassium sorbate. The preservative amount is typically less than about 0.02% (w/v) of the total composition, EDTA may be added up to 2 mg/ml.

[0076] In addition to the ingredients mentioned above, it is contemplated that a variety of additional or alternative ingredients may be present in the pharmaceutical compositions of the present disclosure, which additional or alternative ingredients include anti-oxidants such as vitamin E or its commercially available derivatives such as tocopherol polyethylene glycol 1000 succinate (TPGS), ascorbic acid, or sodium metabisulfite.

[0077] The pharmaceutical compositions herein are typically provided in sterile form for topical administration to the nasal cavity and are preferably adjusted for self-administration by the individual in need thereof. In one embodiment, the preparation is a particle-free nasal spray. Other suitable formulations include intranasally acceptable swabs, as well as ointments and gels that can be applied to the nose, optionally as sprays or aerosols.

[0078] Pharmaceutical compositions described above can be delivered via different methodologies including sprays, irrigation systems (e.g. netipot), syringes or others. The composition may be provided in a dosage form that is suitable for a nasal aerosol or inhalation administration route. An exemplary method of administration of the composition can include spraying vaporized or nebulized disseminated microparticles under an active dynamic pressure.

[0079] Suitable aerosol dispensers for use will be apparent to those skilled in the art and may vary from simple devices analogous to perfume dispensers to pressurized spray cans and even complex apparatus such as might be used in hospitals. Whichever device is used it is generally preferable that it comprises some kind of dosimeter to control the amount of solution administered in one go. One device, which corresponds to a dispenser with a nozzle, effectively incorporates such a dosimeter without any specialized adaptation being necessary, the limit stop of the depressible spray head fixing the maximum single amount of solution dispensable at once. Specially developed spray devices may be made with a hand-held device comprising a reservoir of the composition.

[0080] Suitable means for dispersing the spray, preferably in aerosol form, are provided. Examples include pneumatically pressurized devices and devices employing pressurized gas forced across the opening of a tube leading into the reservoir to create an aerosol, and press-button type devices wherein the button, when pressed, creates pressure on the surface of the liquid in the reservoir, forcing it up through a tube and through a fine nozzle to disperse the solution into an aerosol spray. Other examples include aerosol dispensers, inhalers, pump sprayers, nebulizers (such as positive pressure nebulizers), and the like. In some embodiments, the device used is pre-filled with a composition described herein.

[0081] One embodiment would include a multi-dose metered dose spray pump allowing for spraying of a fixed volume of solution. Alternatively, gas driven (e.g. nitrogen) devices, such as systems that hold the compositions separate from the propellant in aluminum or plastic (or any other type of) bottle. These devices deliver solution at variable diffusion flows and angles when combined with different actuators. Preferred diffusion flows could deliver 0.5-10 ml solution per spraying second at angles of 0-60°.

[0082] The compositions described above can be administered as per physician's instructions and depending on the condition. A preferred mode of (nasal) administration comprises 1-5 sprays per nostril, 1-5 times daily; this could extend to many weeks depending on the condition or symptom to be treated (e.g. in allergy).

[0083] The therapeutic methods described herein (that include prophylactic treatment) in general comprise administration of a therapeutically effective amount of the compositions herein to a subject (e.g., animal, human) in need thereof, including a mammal, particularly a human. Such treatment will be suitably administered to subjects, particularly humans, suffering from, having, susceptible to, or at risk for infection by respiratory tract viruses or symptoms thereof.

[0084] Determination of those subjects “at risk” can be made by any objective or subjective determination by a diagnostic test or opinion of a subject or health care provider.

EXAMPLES

[0085] Examples are provided below to facilitate a more complete understanding of the invention. The following examples illustrate the exemplary modes of making and practicing the invention.

Example 1 In Vitro Inhibition of SARS-CoV-2 Using Iota Carrageenan

[0086] The aim of this study was to determine if Iota Carrageenan demonstrates antiviral activity against SARS-CoV-2.

[0087] An in vitro experiment performed under controlled conditions have demonstrated that even at concentrations of 6 μg/mL iota Carrageenan alone is capable of significantly reducing viral titers of SARS CoV 2 in Vero cell cultures in vitro. A combination of 5% xylitol and iota Carrageenan reduces by at least 5000 times the virus titer in Vero cell cultures in vitro, even using concentrations of iota Carrageenan as low as 0.6 μg/mL. Moreover, 5% xylitol alone reduces by at least 5000 times virus titer in Vero cell cultures in vitro.

[0088] The following table summarizes the formulations tested for antiviral capacity against SARS CoV 2:

TABLE-US-00001 Component Sample 1 Sample 2 Sample 3 Iota Carrageenan 1.7 mg/ml 1.2 mg/ml 1.2 mg/ml Sodium Chloride (NaCl) 0.9% 0.5% 0% Xylitol   0%   0% 5% pH adjusted to 6.00-7.00 6.00-7.00 6.00-7.00

TABLE-US-00002 Component Sample P1 Sample P2 Sample P3 Sodium Chloride (NaOH) 0.9% 0.5% 0% Xylitol   0%   0% 5% pH adjusted to 6.00-7.00 6.00-7.00  6.0-7.00

[0089] The data obtained are summarized below

TABLE-US-00003 TABLE 1 Log reduction of TCID50/mL after 48 hs. (mean of three replicates) Iota Carrageenan Sample 1 Sample 2 Sample 3  600 μg/mL ≥3.75 ≥4.25 ≥4.25   60 μg/mL ≥3.75 ≥4.25 ≥4.25   6 μg/mL 2.45 ≥4.25 ≥4.25  0.6 μg/mL 0.1 0.65 ≥4.25

[0090] Samples P1, P2 and P3 did not contain iota Carrageenan

TABLE-US-00004 Iota Carrageenan Sample P1 Sample P2 Sample P3 0 μg/mL None None ≥4.25

[0091] All data are surprising. However, the antiviral activity of xylitol on its own as shown in the viral titer reduction of Sample P3 is particularly remarkable.

[0092] A detailed description of the test protocols and results follows:

[0093] Materials: [0094] Vero E6 [0095] T-175 tissue culture flask [0096] 0.25% Trypsin-EDTA (1×) [0097] Minimum Essential Medium (MEM) with Earle's salts and L-glutamine, VWR Cat #10-010-CV [0098] Dulbecco's Phosphate Buffered Saline 1× [0099] Fetal bovine serum (FBS), BioWest Cat #S1620, Heat inactivated, (Lot No: 055G17) [0100] Greiner Bio-one TC treated PS sterile cell culture 12 well plates (Cat #07-000-691) [0101] Virus: SARS-CoV-2 UTHSC passage 2, (SARS-Related Coronavirus 2 Isolate USA-WA1/2020)

Samples:

[0102] Sample 1: 600 mcg/mL; 60 mcg/mL, 6 mcg/mL and 0.6 mcg/mL [0103] To obtain the highest concentration a dilution of 7 mL of Sample 1+3 mL of Sample P1 (or bringing to 10 mL with Sample P1) was used. [0104] The stock solution having a concentration of 1200 mcg/mL may be used to obtain the highest final concentration (600 mcg/mL) by applying 0.5 mL in each of the wells tested. [0105] The rest of 1/10 dilutions were obtained using Sample P1 as diluting agent. [0106] All concentrations and control were tested in triplicate. [0107] Sample 2: 600 mcg/mL; 60 mcg/mL, 6 mcg/mL and 0.6 mcg/mL. [0108] To obtain the highest concentration 0.5 mL of Sample 2 was directly applied in each well. [0109] To obtain the 1/10 dilutions, use Sample P2 as diluting agent. [0110] All concentrations tested in triplicate. [0111] Sample 3: 600 mcg/mL; 60 mcg/mL, 6 mcg/mL and 0.6 mcg/mL. [0112] To obtain the highest concentration directly apply 0.5 mL of Sample 3 in each well. [0113] To obtain the 1/10 dilutions, use Sample P3 as diluting agent. [0114] All concentrations tested in triplicate.

[0115] Methodology: [0116] 1) Seed 2.5×10.sup.5 Vero E6 cells to each well in 12-well plate. [0117] 2) Allow cells to grow overnight. No swirling, otherwise cell density will be lower in the center of the well. Cells should be 70-90% confluent the day of infection. 3) Next day remove media, wash each well once with 1 ml PBS [0118] 4) Add: [0119] a) 0.5 ml fresh media (MEM+2% FBS+1% P-S)+0.5 ml Sample 1, 2, or 3 (final concentrations in 1 ml total: 600 mcg/mL; 60 mcg/mL, 6 mcg/mL and 0.6 mcg/mL) [0120] b) 0.5 ml fresh media+0.5 ml Sample P1, P2, or P3 equivalent (Sample equivalent volume amount in μl) [0121] 5) Include virus only and cells only control on the plates [0122] 6) Incubate for 2 hr at 37° C. and then remove well components [0123] 7) Add SARS-CoV-2, 5×10.sup.4/0.2 ml (MOI=0.1) diluted in MEM+2% FBS, to each well. [0124] 8) Incubate for 1 hr at 37° C. Swirl the plates every 15 min [0125] 9) After 1 hr, remove virus, add: [0126] a) 0.5 ml fresh media (MEM+2% FBS+1% P-S)+0.5 ml Sample 1, 2, or 3 (final concentrations in 1 ml total: 600 mcg/mL; 60 mcg/mL, 6 mcg/mL and 0.6 mcg/mL) [0127] b) 0.5 ml fresh media+0.5 ml Sample P1, P2, or P3 equivalent (Sample equivalent volume amount in μl) [0128] 10) Incubate for 2 days at 37° C. [0129] 11) After 2 days, harvest components from each well and perform TCID50 assay.

[0130] TCID.sub.50 Assay:

[0131] Residual virus was measured by TCID50 assay in 96-well plate format with 3 wells per dilution of virus. 10-fold serial dilutions (10.sup.−1 to 10.sup.−7) of collected samples (treated, control, or virus only) were used to infect Vero E6 cells in a 96-well plate. The cell plates were incubated at 37° C., 5% CO.sub.2, with humidity for an additional 2 days. After 2 days, Cell Titer Glo was used to determine viability of cells. The virus endpoint titer was determined using the Reed-Muench formula and expressed as log TCID50/mL.

TABLE-US-00005 TABLE 1 TCID 50/mL Data TCID 50/ml Replicates 1 2 3 Mean Log Reduction Virus Only 1.78E+05 1.78E+05 1.78E+05 1.78E+05 Formulation  600 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 3.75 LOD #1   60 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 3.75 LOD   6 ug/ml 5.62E+02 3.16E+02 1.00E+03 6.26E+02 2.453294082  0.6 ug/ml 1.78E+05 5.62E+04 1.78E+05 1.37E+05 0.112339991 P1 Control   0 ug/ml 3.16E+06 5.62E+06 3.16E+06 3.98E+06

TABLE-US-00006 TABLE 2 TCID50/mL Data TCID 50/ml Replicates 1 2 3 Mean Log Reduction Virus Only 5.62E+05 5.62E+05 5.62E+05 5.62E+05 Formulation  600 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 4.25 LOD #2   60 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 4.25 LOD   6 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 4.25 LOD  0.6 ug/ml 3.16E+05 3.16E+04 3.16E+04 1.26E+05 0.647940009 P2 Control   0 ug/ml 5.62E+05 3.16E+05 5.62E+05 4.80E+05

TABLE-US-00007 TABLE 3 TCID50/mL Data TCID 50/ml Replicates 1 2 3 Mean Log Reduction Virus Only 5.62E+05 5.62E+05 5.62E+05 5.62E+05 Formulation  600 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 4.25 LOD #3   60 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 4.25 LOD   6 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 4.25 LOD  0.6 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 4.25 LOD P3 Control   0 ug/ml 3.16E+01 3.16E+01 3.16E+01 3.16E+01 4.25 LOD

[0132] Effect of Iota Carrageenan on SARS-CoV-2 is shown in FIGS. 1-3. Vero E6 cells were treated with 600 μg/ml, 60 μg/ml, 6 μg/ml or 0.6 μg/ml pre- and post-infection with SARS-CoV-2. After a 2 h pretreatment, cells were infected with SARS-CoV-2 and incubated for 48 h in the presence of IC. Supernatants were harvested and virus yield determined by an end point dilution assay (TCID50). Controls consisted of untreated infected cells or infected cells treated with formulation without IC. Results were determined using the Reed and Muench formula and expressed as TCID50/ml. Dotted line shows the limit of detection (LOD). Testing of samples was performed in triplicate.

Results:

[0133] To examine the antiviral effects of Iota Carrageenan (IC) on SARS-CoV-2, three formulations were developed and tested. Each of the three formulations were tested in a dose dependent manner and ranged from 600 μg/ml to 0.6 μg/ml. The composition of each formulation was as follows:

Formulation #1: SARS-CoV-2 samples treated with 600 μg/ml and 60 μg/ml of IC were reduced ≥3.75 Log when compared to untreated control. The 6 μg/ml concentration of IC also demonstrated an effect but to a lesser extent, with a 2.5 Log reduction in virus. At 0.6 μg/ml, IC did not demonstrate activity and was comparable to the untreated control. Lastly, there was no reduction in virus with P1, providing evidence that IC and not the components of formulation #1 is inhibiting SARS-CoV-2.
Formulation #2: SARS-CoV-2 samples treated with 600 μg/ml, 60 μg/ml, and 6 μg/ml of IC were reduced ≥4.25 Log when compared to untreated control. At 0.6 μg/ml, IC did not demonstrate activity and was comparable to the untreated control. No reduction in virus with P2 was observed, providing evidence that IC and not the components of formulation #2 is inhibiting SARS-CoV-2.
Formulation #3: All concentrations demonstrated antiviral activity including the P3 control. Xylitol was present in this formulation and suggests this component might also exert an antiviral effect.

Example 2

[0134] A nasal spray pharmaceutical formulation comprising:

TABLE-US-00008 Component Amount per 100 mL Function Iota Carrageenan 120 mg Active Ingredient NaCl 0.50 g Active ingredient HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0135] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 8,800 10-mL nasal sprays (an overfill is needed to obtain 10 mL extractable volume):

[0136] 1. Iota carrageenan (120 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0137] 2. NaCl (0.50 kg) is added and dissolved under stirring

[0138] 3. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0139] 4. The solution is brought to volume with additional Purified Water

[0140] 5. The final solution is sterilized by filtration using a suitable filtration device.

[0141] 6. Sterile solution is aseptically filled into bottles

[0142] 7. Preservative-free metered nasal pumps are snapped onto the bottle necks

[0143] 8. Units are labeled and packaged for distribution

[0144] In this preparation Purified Water may be replaced by Water for Injection.

Example 3

[0145] A nasal spray pharmaceutical formulation comprising:

TABLE-US-00009 Amount per 100 Component mL Function Iota Carrageenan 170 mg Active Ingredient NaCl 0.90 g Active ingredient HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0146] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 8,800 10-mL nasal sprays (an overfill is needed to obtain 10 mL extractable volume):

[0147] 1. Iota carrageenan (170 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0148] 2. NaCl (0.90 kg) is added and dissolved under stirring

[0149] 3. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0150] 4. The solution is brought to volume with additional Purified Water

[0151] 5. The final solution is sterilized by filtration using a suitable filtration device.

[0152] 6. Sterile solution is aseptically filled into bottles

[0153] 7. Preservative-free metered nasal pumps are snapped onto the bottle necks

[0154] 8. Units are labeled and packaged for distribution

[0155] In this preparation Purified Water may be replaced by Water for Injection.

Example 4

[0156] A nasal spray pharmaceutical formulation comprising:

TABLE-US-00010 Component Amount per 100 mL Function Iota Carrageenan 120 mg Active Ingredient Xylitol 7.00 g Active ingredient HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0157] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 8,800 10-mL nasal sprays (an overfill is needed to obtain 10 mL extractable volume):

[0158] 1. Iota carrageenan (120 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0159] 2. Xylitol (7 kg) is added and dissolved under stirring

[0160] 3. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0161] 4. The solution is brought to volume with additional Purified Water

[0162] 5. The final solution is sterilized by filtration using a suitable filtration device.

[0163] 6. Sterile solution is aseptically filled into bottles

[0164] 7. Preservative-free metered nasal pumps are snapped onto the bottle necks

[0165] 8. Units are labeled and packaged for distribution

[0166] In this preparation Purified Water may be replaced by Water for Injection.

Example 5

[0167] A nasal spray pharmaceutical formulation comprising:

TABLE-US-00011 Amount per 100 Component mL Function Xylitol 0.50 g Active ingredient NaCl 0.81 g Osmolyte HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0168] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 8,800 10-mL nasal sprays (an overfill is needed to obtain 10 mL extractable volume):

[0169] 1. Xylitol (0.5 kg) is added and dissolved under stirring

[0170] 2. NaCl (0.81 kg) is added and dissolved under stirring

[0171] 3. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0172] 4. The solution is brought to volume with additional Purified Water

[0173] 5. The final solution is sterilized by filtration using a suitable filtration device.

[0174] 6. Sterile solution is aseptically filled into bottles

[0175] 7. Preservative-free nasal metered pumps are crimped onto the bottle necks

[0176] 8. Units are labeled and packaged for distribution

[0177] In this preparation Purified Water may be replaced by Water for Injection.

Example 6

[0178] A nasal spray pharmaceutical formulation comprising:

TABLE-US-00012 Amount per 100 Component mL Function Iota Carrageenan 170 mg Active Ingredient NaCl 0.90 g Active ingredient Potassium Sorbate 0.20 g Preservative Disodium Edetate 0.10 g Chelating agent HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0179] A pharmaceutical formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 8,800 10-mL nasal sprays (an overfill is needed to obtain 10 mL extractable volume):

[0180] 1. Iota carrageenan (170 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0181] 2. NaCl (0.9 kg) is added and dissolved under stirring

[0182] 3. Potassium Sorbate (0.20 kg) is added and dissolved under stirring

[0183] 4. Disodium Edetate (0.10 kg) is added and dissolved under stirring.

[0184] 5. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0185] 6. The solution is brought to volume with additional Purified Water

[0186] 7. The final solution is filled into bottles.

[0187] 8. Metered nasal pumps are snapped onto the bottle necks

[0188] 9. Units are labeled and packaged for distribution

Example 7

[0189] A nasal spray pharmaceutical formulation comprising:

TABLE-US-00013 Amount per 100 Component mL Function Iota Carrageenan 120 mg Active Ingredient Xylitol 5.00 g Active ingredient Potassium Sorbate 0.20 g Preservative Disodium Edetate 0.10 g Chelating agent HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0190] A pharmaceutical formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 8,800 10-mL nasal sprays (an overfill is needed to obtain 10 mL extractable volume):

[0191] 1. Iota Carrageenan (120 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0192] 2. Xylitol (5 kg) is added and dissolved under stirring

[0193] 3. Potassium Sorbate (0.20 kg) is added and dissolved under stirring

[0194] 4. Disodium Edetate (0.10 kg) is added and dissolved under stirring.

[0195] 5. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0196] 6. The solution is brought to volume with additional Purified Water

[0197] 7. The final solution is filled into bottles.

[0198] 8. Metered nasal pumps are crimped onto the multidose bottles

[0199] 9. Units are labeled and packaged for distribution

Example 8

[0200] A nasal spray pharmaceutical formulation comprising:

TABLE-US-00014 Amount per 100 Component mL Function Iota Carrageenan 170 mg Active Ingredient Xylitol 0.50 g Active ingredient NaCl 0.81 g Osmolyte Potassium Sorbate 0.20 g Preservative Disodium Edetate 0.10 g Chelating agent HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0201] A pharmaceutical formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 8,800 10-mL nasal sprays (an overfill is needed to obtain 10 mL extractable volume):

[0202] 1. Iota Carrageenan (170 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0203] 2. Xylitol (5 kg) is added and dissolved under stirring

[0204] 3. Potassium Sorbate (0.20 kg) is added and dissolved under stirring

[0205] 4. Disodium Edetate (0.10 kg) is added and dissolved under stirring.

[0206] 5. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0207] 6. The solution is brought to volume with additional Purified Water

[0208] 7. The final solution is filled into bottles.

[0209] 8. Metered nasal pumps are snapped onto the bottles

[0210] 9. Units are labeled and packaged for distribution

Example 9

[0211] A pharmaceutical formulation for either nasal or lung nebulization comprising:

TABLE-US-00015 Amount per 100 Component mL Function Iota Carrageenan 50.0 mg Active Ingredient NaCl 0.90 g Active ingredient HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0212] This formulation contains 1 mg each 2 ml to be nebulized into both nostrils or into the lungs in a dose similar to the nasal sprays. NaCl is formulated at 0.90%.

[0213] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca 47,600 2-mL unit dose vials (an overfill is needed to obtain 3 mL extractable volume):

[0214] 1. Iota Carrageenan (50 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0215] 2. NaCl (0.90 kg) is added and dissolved under stirring

[0216] 3. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0217] 4. The solution is brought to volume with additional Purified Water

[0218] 5. The final solution is sterilized by filtration using a suitable filtration device.

[0219] 6. Sterile solution is aseptically filled into unit dose vials.

[0220] 7. Vials are packaged for distribution

[0221] In this preparation Purified Water may be replaced by Water for Injection.

Example 10

[0222] A pharmaceutical formulation for either nasal or lung nebulization comprising:

TABLE-US-00016 Amount per 100 Component mL Function Iota Carrageenan 100.0 mg Active Ingredient Xylitol 5.00 g Active ingredient HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0223] This formulation contains 2 mg each 2 ml to be nebulized into both nostrils or into the lungs in a dose similar to the nasal sprays. Xylitol is formulated at 5%.

[0224] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca 47,600 2-mL unit dose vials (an overfill is needed to obtain 3 mL extractable volume):

[0225] 1. Iota Carrageenan (100 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0226] 2. Xylitol (5 kg) is added and dissolved under stirring

[0227] 3. Either HCl or NaOH is added to reach pH between 6 and 7.5.

[0228] 4. The solution is brought to volume with additional Purified Water

[0229] 5. The final solution is sterilized by filtration using a suitable filtration device.

[0230] 6. Sterile solution is aseptically filled into unit dose vials.

[0231] 7. Vials are packaged for distribution

[0232] In this preparation Purified Water may be replaced by Water for Injection.

Example 11

[0233] A pharmaceutical formulation for either nasal or lung nebulization comprising:

TABLE-US-00017 Amount per 100 Component mL Function Iota Carrageenan 50.0 mg Active Ingredient Xylitol 0.50 g Active ingredient NaCl 0.81 g Osmolyte HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0234] This formulation contains 1 mg each 2 ml to be nebulized into both nostrils or into the lungs in a dose similar to the nasal sprays. Xylitol and NaCl are formulated at 0.50% and 0.81% w/v.

[0235] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca 47.600 2-mL unit dose vials (an overfill is needed to obtain 3 mL extractable volume):

[0236] 1. Iota Carrageenan (50.0 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0237] 2. Xylitol (5 kg) is added and dissolved under stirring

[0238] 3. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0239] 4. The solution is brought to volume with additional Purified Water.

[0240] 5. The final solution is sterilized by filtration using a suitable filtration device.

[0241] 6. Sterile solution is aseptically filled into unit dose vials.

[0242] 7. Vials are packaged for distribution

[0243] In this preparation Purified Water may be replaced by Water for Injection.

Example 12

[0244] A pharmaceutical formulation for either nasal or lung nebulization comprising:

TABLE-US-00018 Amount per 100 Component mL Function Iota Carrageenan 17.0 mg Active Ingredient Xylitol 5.00 g Active ingredient Potassium Sorbate 0.20 g Preservative Disodium Edetate 0.10 g Chelating agent HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0245] This formulation contains 0.34 mg of iota carrageenan each 2 ml to be nebulized into both nostrils or into the lungs in a dose similar to the nasal sprays. Xylitol is formulated at 5.0%.

[0246] A pharmaceutical formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 3,200 multidose 30-mL bottles (an overfill is needed to obtain 10 mL extractable volume):

[0247] 1. Iota Carrageenan (17.0 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0248] 2. Xylitol (5 kg) is added and dissolved under stirring

[0249] 3. Potassium Sorbate (0.20 kg) is added and dissolved under stirring

[0250] 4. Disodium Edetate (0.10 kg) is added and dissolved under stirring.

[0251] 5. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0252] 6. The solution is brought to volume with additional Purified Water.

[0253] 7. The solution is filled into multidose bottles

[0254] 8. Bottles are packaged for distribution

Example 13

[0255] A pharmaceutical formulation for either nasal or lung nebulization comprising:

TABLE-US-00019 Amount per 100 Component mL Function Iota Carrageenan 17.0 mg Active Ingredient Xylitol 0.50 g Active ingredient NaCl 0.81 g Osmolyte Potassium Sorbate 0.20 g Preservative Disodium Edetate 0.10 g Chelating agent HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0256] This formulation contains 0.34 mg each 2 ml to be nebulized into both nostrils or into the lungs in a dose similar to the nasal sprays. Xylitol and NaCl are formulated at 0.50% and 0.81% w/v.

[0257] A pharmaceutical formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 8,800 10-mL nasal sprays (an overfill is needed to obtain 10 mL extractable volume):

[0258] 1. Iota Carrageenan (17.0 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0259] 2. Xylitol (5 kg) is added and dissolved under stirring

[0260] 3. NaCl (0.81 kg) is added and dissolved under stirring

[0261] 4. Potassium Sorbate (0.20 kg) is added and dissolved under stirring

[0262] 5. Disodium Edetate (0.10 kg) is added and dissolved under stirring.

[0263] 6. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0264] 7. The solution is brought to volume with additional Purified Water.

[0265] 8. The solution is filled into multidose bottles

[0266] 9. Bottles are packaged for distribution

Example 14

[0267] A pharmaceutical formulation for nasal lavage or nasal wash:

TABLE-US-00020 Amount per 100 Component mL Function Iota Carrageenan 100.0 mg Active Ingredient Xylitol 5.00 g Active ingredient HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0268] This formulation contains 2 mg each 2 ml to be used as a nasal rinse. Xylitol is formulated at 5%.

[0269] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 1,500 60-mL units in multidose sprays or pressurized aerosols (an overfill is needed to obtain 3 mL extractable volume):

[0270] 1. Iota Carrageenan (100 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0271] 2. Xylitol (5 kg) is added and dissolved under stirring

[0272] 3. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0273] 4. The solution is brought to volume with additional Purified Water

[0274] 5. The final solution is sterilized by filtration using a suitable filtration device.

[0275] 6. Sterile solution is aseptically filled either into multidose spray bottles or aerosol cans.

[0276] 7. In case of spray bottles a suitable pump is crimped, snapped or screw capped onto the. In the case of aerosol cans a suitable continuous spray valve is crimped onto it.

[0277] In this preparation Purified Water may be replaced by Water for Injection.

Example 15

[0278] A pharmaceutical formulation for nasal lavage or nasal wash:

TABLE-US-00021 Amount per 100 Component mL Function Iota Carrageenan 12.0 mg Active Ingredient Xylitol 5.00 g Active ingredient Potassium Sorbate 0.20 g Preservative Disodium Edetate 0.10 g Chelating agent HCl/NaOH s.q. pH 5-7.5 pH adjustment agents Water s.q. 100 mL Vehicle

[0279] This formulation was manufactured in the following way to manufacture 100 L of bulk solution, yielding ca. 1,500 60-mL units in multidose sprays or pressurized aerosols (an overfill is needed to obtain 3 mL extractable volume):

[0280] 1. Iota carrageenan (12.0 g) is dissolved in 90 Liter of Purified Water under stirring in a mixing tank.

[0281] 2. Xylitol (5 kg) is added and dissolved under stirring

[0282] 3. Potassium Sorbate (0.20 kg) is added and dissolved under stirring

[0283] 4. Disodium Edetate (0.10 kg) is added and dissolved under stirring.

[0284] 5. Either HCl or NaOH is added to reach pH between 6 and 7.5

[0285] 6. The solution is brought to volume with additional Purified Water

[0286] 7. In case of spray bottles a suitable pump is crimped, snapped or screw capped onto the top. In the case of aerosol cans a suitable continuous spray valve is crimped onto it.

Example 16

[0287]

TABLE-US-00022 Candidate formulations for nasal sprays to prevent transmission and alleviate symptoms and reduce viral load at an early stage of COVID 19 Component Amount per 100 mL Function Iota Carrageenan 170 mg Active Ingredient NaCl 0.90 g Active ingredient HCl/NaOH s.q. pH 6-7 pH adjustment agents Water s.q. 100 mL Vehicle Dose to be applied 100 μL in each nostril each 4 hours or prior to exposure to COVID 19 patients (i.e. 170 μg of iota Carrageenan in each nostril) Iota Carrageenan 120 mg Active Ingredient NaCl 0.50 g Active ingredient HCl/NaOH s.q. pH 6-7 pH adjustment agents Water s.q. 100 mL Vehicle Dose to be applied 140 μL or 100 μL in each nostril each 4 hours or prior to exposure to COVID 19 patients (i.e. 168 μg or 120 μg of iota Carrageenan in each nostril) Iota Carrageenan 170 mg Active Ingredient Xylitol 5 g Active ingredient HCl/NaOH s.q. pH 6-7 pH adjustment agents Water s.q. 100 mL Vehicle Dose to be applied 100 μL in each nostril each 4 hours or prior to exposure to COVID 19 patients (i.e. 170 μg of iota Carrageenan in each nostril) Iota Carrageenan 120 mg Active Ingredient Xylitol 5 g Active ingredient HCl/NaOH s.q. pH 6-7 pH adjustment agents Water s.q. 100 mL Vehicle Dose to be applied 140 μL or 100 μL in each nostril each 4 hours or prior to exposure to COVID 19 patients (i.e. 168 μg or 120 μg of iota Carrageenan in each nostril) Iota Carrageenan 170 mg Active Ingredient NaCl 0.90 g Active ingredient Potassium Sorbate 0.20 g Preservative Disodium Edetate 0.10 g Chelating agent HCl/NaOH s.q. pH 5-6 pH adjustment agents Water s.q. 100 mL Vehicle Dose to be applied 100 μL in each nostril each 4 hours or prior to exposure to COVID 19 patients (i.e. 170 μg of iota Carrageenan in each nostril)

Example 17

[0288] Certain formulations described herein comprise 0.01-0.5% w/v iota Carrageenan alone or 0.001-0.5% w/v iota Carrageenan+0.5-10% w/v Xylitol. The formulations are delivered to a subject in 50-200 microliters of formulation per shot.

This would mean:
One shot of iota Carrageenan alone formulation would contain:

[0289] 5 micrograms of iota Carrageenan (0.01% and 50 microliter) to 1000 micrograms or 1 mg of iota Carrageenan (0.5% and 200 microliter)

One shot of iota Carrageenan+Xylitol would contain:

[0290] 0.5 micrograms of iota Carrageenan (0.001% and 50 microliter) to 1000 micrograms or 1 mg of iota Carrageenan (0.5% and 200 microliter).

[0291] 250 micrograms of Xylitol (0.5% and 50 microliter) to 20 mg (5% and 200 microliter) Each application is one shot in each nostril, i.e. that the dose is double these amounts as follows:

Two shots of iota Carrageenan alone formulation would contain:

[0292] 10 micrograms of iota Carrageenan (0.01% and 50 microliter) to 2000 micrograms or 2 mg of iota Carrageenan (0.5% and 200 microliter).

Two shots of iota Carrageenan+Xylitol would contain:

[0293] 1 microgram of iota Carrageenan (0.001% and 50 microliter) to 2000 micrograms or 2 mg of iota Carrageenan (0.5% and 200 microliter).

[0294] 500 micrograms of Xylitol (0.5% and 50 microliter) to 40 mg (5% and 200 microliter).

OTHER EMBODIMENTS

[0295] While the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0296] The patent and scientific literature referred to herein establishes the knowledge that is available to those with skill in the art. All United States patents and published or unpublished United States patent applications cited herein are incorporated by reference. All published foreign patents and patent applications cited herein are hereby incorporated by reference. All other published references, documents, manuscripts and scientific literature cited herein are hereby incorporated by reference.

[0297] While this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.