Compositions and uses thereof

Abstract

A dry powder composition for administration to the nasal cavity is provided. [Solution] The composition consists of only two or more ingredients: i) hydroxypropyl methylcellulose particles; and ii) at least one chemical agent selected from signal transduction agents, wherein the particles of the homogenized dry powder have an average particle size in the range of 80 μm to 101 μm, the signal transduction agent includes garlic, and the composition is for use as a nasally administered pharmaceutical.

Description

[Technical Field]

[0001] The present invention relates to dry powder compositions for administration to the nasal cavity and uses thereof. In particular, the compositions of the present invention comprise powdered hydroxypropyl methylcellulose (pHPMC) particles of a defined average particle size, a signaling agent, and optionally a biologically active agent. [Background technology]

[0002] Allergic rhinitis (AR) is a global health problem affecting up to 25% of the adult population and over 40% of children in developed countries and is thought to be responsible for an annual economic loss of approximately $2 billion to $5 billion in the United States alone. 1 .

[0003] It is known, for example from U.S. Patent No. 8,202,550, that HPMC powder and the gel it forms when administered to the nasal cavity are an effective means of intranasally administering therapeutic agents, particularly herbal or homeopathic agents. This prior art invention teaches that the therapeutic agents contained in the compositions described therein provide a therapeutic effect. When administered to the nasal cavity, pHPMC forms a protective gel that slowly releases moisture upon contact with the nasal mucosa, and any co-administered agents. Reported for 6 hours 2 In comparison, administration of an active agent to the nasal cavity in a conventional intranasal composition (such as a liquid or powder composition containing little or no cellulose) typically provides a relatively short duration (less than 4 hours) of therapeutic effect. However, there is no disclosure of average particle size in U.S. Patent No. 8,202,550.

[0004] The prior art further teaches or suggests the use of HPMC powder compositions to treat allergic rhinitis (AR) and other respiratory illnesses (including certain viral illnesses, although there is no data regarding coronavirus illnesses, particularly respiratory illnesses associated with the SARS-CoV-2 virus). However, the prior art is silent about the average particle size of the powdered HPMC prior to formation into a gel.

[0005] Diethart B et al., Nat. Sci. 2010; Vol. 2, No. 2: 79-84, teaches the possibility of using HPMC to block the diffusion of house dust mite allergens through HPMC and agar gels, thus blocking allergen uptake. 3 No mention is made of the average particle size of the powdered HPMC prior to formation into a gel.

[0006] A Russian study by MKErofeeva et al. (https: / / medi.ru / info / 7023 / ) indicates that HPMC is useful in preventing influenza in children. 4 However, there was no indication of the mean particle size used in the dry powder HPMC-containing formulations used in the study.

[0007] Defective nasal barrier function contributes to allergic rhinitis, resulting in persistent inflammation and clinical symptoms, with congestion playing a prominent role. A recent study by Valerieva, A et al. in Allergy Asthma Proc 36:1-6, 2015; doi:10.2500 / aap.2015.36.3879 showed that administration of HPMC followed by oxymetazoline provided relief in a sample of patients known to be sensitive to at least one of a range of allergens across four seasons. 2 The authors stated that further research is needed due to the statistical methods used, the short study period, and the small sample size. There was no clear mention of average particle size in this study, and it does not appear that signaling agents were used.

[0008] A paper by Popov TA et al. on HPMC powder for the prevention and management of nasal symptoms (Popov TA et al., Expert Review of Respiratory Medicine, 2017 Vol. 11, No. 11, 885-892 (https: / / doi.org / 10.1080 / 17476348.2017.1375408) reported that HPMC provides a natural barrier against pollen allergens and harmful agents. 5 The paper itself and some of the studies cited therein included a variety of plant pollen and house dust mite allergens, but did not mention the average particle size of the powder preparations used.

[0009] HPMC powder by itself and HPMC powder containing peppermint and wild garlic have been shown to reduce H5N1 viral titers in SPEV cell cultures in in vitro studies when compared to controls (Lvov DK and Deryabin PG, "Virucidal Activity of Nasaleze (Nasaval) and Nasaleze Travel (Nasaleze Plus) in Cell Cultures Infected with Pathogenic Avian Flu Virus (H5N1)," 2010, European Journal for Nutraceutical Research 1-8, www.phytomedcentral.org). 6 There is no mention of the average particle size and how it may affect the effectiveness of the HPMC powder as a blocking agent against viral uptake.

[0010] The addition of HPMC powder and stabilized allicin extract from garlic was shown in the evaluation of a specific strain of methicillin-resistant Staphylococcus aureus, namely UEL301, and showed significant biological activity in various formulations (Cutler University of East London, March 2003). There is no mention of the average particle size and how it may affect the effectiveness of HPMC powder as a blocking agent for bacterial removal.

[0011] Powdered compositions of HPMC containing only a signaling agent and / or biologically active agent having the average particle size described herein and capable of forming a gel upon contact with moisture have been shown to be more effective than prior art compositions in reducing allergic rhinitis (AR) caused by airborne particles such as viral infections (virus particles size range 0.005 μm to 0.3 μm), pollen grains (size range 10 μm to 1000 μm), allergic reactions caused by inhalation of particles from biological sources such as house dust mites (size range 100 μm to 300 μm), and PM 2.5 From PM 10 etc., have been found to be more efficient in controlling or containing allergic reactions caused by inhalation of airborne pollutant particles (size range 1 μm to 150 μm). 7 This is surprising given the wide range of inhaled particle sizes and the range of average particle sizes of the inventive HPMC powders.

[0012] Given two recent outbreaks of deadly coronavirus disease in the Middle East, and a third outbreak in the Middle East where a new coronavirus strain that infects humans is currently evolving, there is an urgent need to provide formulations that can at least slow, block, and / or neutralize the spread of such viruses before they become established in the host organism. A further problem with the new coronavirus strain from the Far East (2019-n CoV, also known as SAR-COV-2) is that it can persist in the host (human) for approximately two weeks before symptoms appear. When the compositions of the present invention are applied prophylactically and / or curatively to the nasal lining via regular insufflation, disease containment may be possible.

[0013] The above-suggested advantages and other advantages will become apparent from the following description.

[0014] Previously, it was thought that the average particle size should be in the range of 5 μm to 500 μm, but new data, as detailed herein, indicates that the optimum average particle size is in the range of 20 μm to 500 μm.

[0015] It has now been found that mean particle sizes of the dry powder compositions of the present invention in the range of about 20 μm to about 500 μm, preferably in the range of 60 μm to 150 μm, and more preferably in the range of 80 to 125 μm, are surprisingly effective in trapping airborne allergens in the gel. Furthermore, the dry powder compositions of the present invention designed for intranasal application are promising for the delivery of drugs against airborne viruses such as coronaviruses. Summary of the Invention

[0016] According to the present invention, i) hydroxypropyl methylcellulose particles, and ii) at least one chemical agent selected from signaling agents, and / or iii) one or more biologically active agents; A composition in the form of a homogenized dry powder consisting of two or more components selected from The homogenized dry powder particles have an average particle size of 20 μm or more and 500 μm or less.

[0017] The powder compositions of the present invention preferably have an average particle size in the range of 20 to 500 μm, preferably 60 to 150 μm, more preferably 80 to 125 μm, for example 86 μm + / - 15 μm, depending on the signalling agent added and / or the biologically active agent added.

[0018] The compositions of the present invention are designed for application to the nasal mucosa by insufflation through the nose.

[0019] The compositions of the present invention must be capable of forming a gel upon contact with moisture, as shown in the accompanying examples. The compositions of the present invention should not contain additives that could interfere with or substantially interfere with their ability to form a gel upon contact with moisture, such as additives that could significantly lower the pH of the nasal mucosa. The dry powder particles of the present invention absorb moisture upon contact with the nasal mucosa, thereby forming a gel matrix on their surface. The function of the gel is believed to be at least twofold: first, it acts as a physical barrier against the uptake of small particles, such as airborne allergens and viruses, through the nasal mucosa, and second, it allows the selected drug to diffuse across the nasal mucosa cells and into the bloodstream. During hydration of the dry powder compositions of the present invention, a gel matrix is ​​formed upon contact with moisture, with larger and smaller particles combining to form a molecular net or matrix, with the smaller particles occupying the spaces or interstices between the larger particles, thus contributing to gel formation and helping the larger particles to be more easily enclosed together. Particulate matter is trapped in the gel and is largely unable to pass through the mucosa.

[0020] The compositions of the present invention may include biologically active agents selected from pharmaceutical agents, herbal agents, and homeopathic agents. Suitable homeopathic and herbal agents may be selected from St. John's wort, valerian extract, ginkgo biloba extract, vitamins A, E or C, garlic, lime, one or more probiotics, ginger, ellagic acid, echinacea, Swedish flower pollen, black walnut shells, lemongrass, mugwort, grapefruit seed extract, broccoli, digestive enzymes, hyaluronic acid, astragalus, rose hips, gentian, hypericum, horse chestnut, ginseng, green tea, phosphatidylserine, phosphatidylcholine, citrus fruits, pycnogenol, caffeine, quercetin, coenzyme Q10, yarrow, tea tree, noni juice, lipase, fructooligosaccharides, inulin, black cumin, stabilized allicin, or any combination thereof.

[0021] The compositions of the present invention may comprise a biologically active agent selected from pharmaceutical antiviral agents, i.e., Type I (α, β) interferons (IFNs), such as IFN-β, IFNβ-1b, Type II (γ) and Type III (λ) interferons, favipiravir (also known as Favilovir, T-705, and Avigan) available from Fujifilm Toyama Chemica, remdesivir, ozeltamivir, zanamivir, ribavirin, lopinavir, the combination of lopinavir-ritonavir with IFNβ-1b, monoclonal and (camelid) polyclonal neutralizing antibodies, and macrolides, such as ivermectin, plant alkaloids, such as colchicine, etc. A compound that generally shows potential for use against coronaviruses is K22, structural name (Z)-N-(3-(4-(4-bromophenyl)-4-hydroxypiperidin-1-yl)-3-oxo-1-phenylprop-1-en-2-yl)benzamide, available from ChemDiv (San Diego, CA, catalog number 4295-0370). This compound targets membrane-bound viral RNA synthesis and shows potent inhibition in various coronaviruses, including MERS. Further suitable biologically active agents include the isolated griffithsin lectin protein of approximately 121 amino acids extractable from red algae such as Griffithsia, and its biologically active isolated antiviral analogs, and natural seaweed extracts containing same (Journal of Virology 2010, O'Keefe, BR et al., "Broad Spectrum In Vitro Activity and In Vivo Efficacy of the Antiviral Protein Griffithsin against Emerging Viruses of the Family Coronaviridae"). 8.Published online DOI: 10.1128 / JVI.02322-09; Marine Drugs 2019 Oct.;17(10):567, Choongho Lee, "Griffithsin - a highly potent broad spectrum antiviral lectin from red algae: From discovery to clinical application," published online October 6, 2019 doi:10.3390 / md 17100567.

[0022] The compositions according to the present invention may contain pHPMC, a biologically active agent as defined herein, and a signaling agent or additive, such as menthol, strawberry, mint, spearmint, peppermint, eucalyptus, lavender, and citrus fruits, or any combination thereof. Examples of citrus fruits may include lemon, lime, and kumquat. Preferably, the signaling agent is one that is not known to be or has not been implicated as an irritant to the nasal mucosa, such as one selected from lemon, lime, kumquat, and strawberry.

[0023] The signaling agent may be present at 0.25% w / w to ≦2% w / w, preferably 0.50% w / w to 2% w / w of the total weight of the composition. The biologically active agent comprises 8% w / w to 9.75% w / w of the composition. The homogenized dry powder compositions of the present invention consist of 90% w / w up to 99.75% w / w of HPMC particles, depending on the design.

[0024] The compositions of the present invention are effective against viruses selected from influenza viruses, such as influenza A, H1N1, H5N1 and H3N2, coronaviruses, such as MERS-CoV, SAR-CoV, HCoV-229E, HCoV-NL63, HCoV-OC43, CoV-HKU1, and 2019nCoV (also known as SAR-COV-2), and bacteria, such as Staphylococcus aureus, methicillin resistant Staphylococcus aureus, Haemophilus influenzae, Mycobacterium tuberculosis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecium, and the like. The present invention can physically contain and / or destroy the physiology of bacteria such as Candida faecium, Candida albicans, Candida tropicalis, and Enterobacter species.

[0025] In a second embodiment of the present invention, i) hydroxypropyl methylcellulose particles; ii) at least one chemical agent selected from signaling agents; A composition in the form of a homogenized dry powder is provided, comprising:

[0026] The homogenized dry powder (pHPMC) of the second embodiment of the present invention has the same defined average particle size as provided above.

[0027] The composition according to the second embodiment of the present invention contains a signaling agent or additive, such as menthol, strawberry, mint, spearmint, peppermint, eucalyptus, lavender, and citrus fruits, or any combination thereof. Examples of citrus fruits include lemon, lime, and kumquat. Preferably, the signaling agent is not known to be or is not associated with nasal mucosa irritation, such as lemon, lime, kumquat, and strawberry.

[0028] The signaling agent in the second embodiment of the invention comprises 0.25% to 10% w / w or less, preferably 0.50% to 5% w / w, of the total weight of the composition. The homogenized dry powder composition of the second embodiment of the invention comprises from about 90% w / w to up to 99.75% w / w of HPMC particles as defined herein, depending on design. Preferably, the composition of the second embodiment of the invention contains more than 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% w / w of the composition, depending on design.

[0029] The inclusion of such signaling agents in the compositions of the present invention is intended to provide the patient with sensory feedback upon use in the form of a distinct sensation that allows the patient to recognize that administration has occurred and can aid in the patient's memory of administration. Thus, for purposes of the present invention, signaling agents are defined as those that primarily impart olfactory and / or gustatory sensations to the user.

[0030] Signaling agents may have other beneficial effects on subjects.Without intending to be bound by theory, certain formulations according to the present invention that contain mint may have the effect of helping to dilate airways.This may be particularly beneficial when the formulation is used to treat patients suffering from asthma.Some patients, especially those under stress, tend to breathe in an irregular pattern.Administering HPMC formulations that contain agents such as mint may also provide a feel-good factor that can help restore normal breathing patterns.

[0031] In a third embodiment of the present invention, i) hydroxypropyl methylcellulose particles; ii) one or more biologically active agents selected from antiviral agents, antibacterial agents, and antiallergic agents; A composition comprising:

[0032] The biologically active agent may comprise 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% w / w of the composition, or any value therebetween, depending on the design. The homogenized dry powder composition of the third embodiment of the present invention comprises 90% w / w to 98% w / w of pHPMC particles. Preferably, the composition of the third embodiment of the present invention contains pHPMC particles as defined herein at greater than 90, 91, 92, 93, 94, 95, 96, 97, or 98% w / w of the composition, or any value therebetween, depending on the design.

[0033] The composition of the third embodiment of the present invention comprises one or more biologically active agents selected from therapeutic agents selected from pharmaceuticals, herbal agents, and homeopathic agents. Suitable herbal and homeopathic agents are typically selected from those having one or more of the following functions: antibacterial, antiviral, or anti-inflammatory. The selection of biologically active agents for use in the third aspect of the present invention is St. John's wort, valerian extract, ginkgo biloba extract, vitamins A, E or C, garlic, one or more probiotics, ginger, ellagic acid, echinacea, Swedish flower pollen, black walnut shells, lemongrass, mugwort, grapefruit seed extract, broccoli, digestive enzymes, hyaluronic acid, astragalus, rose hips, gentian, hypericum, horse chestnut, ginseng, green tea, phosphatidylserine, phosphatidylcholine, pycnogenol, caffeine, quercetin, coenzyme Q10, yarrow, tea tree, noni juice, lipase, fructooligosaccharides, inulin, black cumin, stabilized allicin, or any combination thereof.

[0034] Further suitable biologically active agents for use in the third embodiment of the present invention are pharmaceuticals that show promise against diseases caused by coronaviruses, such as Severe Acute Respiratory Syndrome (SARS), COVID19 caused by the SAR-COV2 virus, Middle East Respiratory Syndrome (MERS), coronavirus E229E, and variants thereof selected from type I (α, β) interferons (IFN), such as IFN-β, IFNβ-1 b, type II (γ) and type III (λ) interferons, favipiravir (also known as Favilovir, T-705, and Avigan) available from Fujifilm Toyama Chemica, remdesivir, ozeltamivir, zanamivir, ribavirin, lopinavir, the combination of lopinavir-ritonavir with IFNβ-1 b, monoclonal and (camelid) polyclonal neutralizing antibodies, and macrolides, such as ivermectin, plant alkaloids, such as colchicine. A compound that generally shows potential for use against coronaviruses is K22, structural name (Z)-N-(3-(4-(4-bromophenyl)-4-hydroxypiperidin-1-yl)-3-oxo-1-phenylprop-1-en-2-yl)benzamide, available from ChemDiv (San Diego, CA, catalog number 4295-0370). This compound targets membrane-bound viral RNA synthesis and shows potent inhibition in various coronaviruses, including MERS.Further suitable biologically active agents include the isolated griffithsin lectin protein of approximately 121 amino acids extractable from red algae such as Griffithsia, and its biologically active isolated antiviral analogs, and natural seaweed extracts containing same (Journal of Virology 2010, O'Keefe, BR et al. "Broad Spectrum In Vitro Activity and In Vivo Efficacy of the Antiviral Protein Griffithsin against Emerging Viruses of the Family Coronaviridae." Published online DOI: 10.1128 / JVI.02322-09). 8 ;Marine Drugs 2019 Oct.;17(10):567, Choongho Lee, "Griffithsin - A Highly Potent Broad Spectrum Antiviral Lectin from Red Algae: From Discovery to Clinical Application," published online October 6, 2019, doi:10.3390 / md 17100567.

[0035] Preferred medications include remdesivir and ivermectin.

[0036] Typically, the compositions of the present invention provide sustained release of a biologically active agent as defined herein. Typically, the biologically active agent has a systemic effect when administered intranasally.

[0037] In certain embodiments of the present invention, a combination of HPMC, a signaling agent, and a biologically active agent is provided for sequential or simultaneous administration. The HPMC, signaling agent, and biologically active agent may be contained together in a single preparation. Alternatively, the HPMC, signaling agent, and biologically active agent may be provided in separate preparations for sequential administration. When administration is sequential, the HPMC and / or signaling agent may be administered before or after the biologically active agent, or both. Similarly, the biologically active agent may be administered before or after the HPMC and / or signaling agent, or both.

[0038] When the powdered HPMC and / or signaling agent are included in the same preparation as the biologically active agent, the preparation is preferably in powder form. When the powdered HPMC and / or signaling agent are included in a separate preparation from the biologically active agent, the HPMC is preferably in powder form. However, the biologically active agent may be in any form, preferably in a form suitable for nasal administration, such as powder, liquid, cream, or gel form.

[0039] In the compositions of the present invention, the powdered HPMC is present in a proportion of at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% w / w of the total weight of the composition, depending on the design.

[0040] In another embodiment of the present invention, a combination of HPMC and a biologically active agent is provided for sequential or simultaneous administration. The HPMC and the biologically active agent may be contained together in a single preparation. Alternatively, the HPMC and the biologically active agent may be provided in separate preparations for sequential administration. When administration is sequential, the HPMC may be administered before and / or after the biologically active agent. Alternatively, the biologically active agent may be administered before and / or after the HPMC.

[0041] When the HPMC and biologically active agent are contained in separate preparations, the agent may be in any form suitable for intranasal administration, such as a powder, liquid, cream, or gel.

[0042] According to one embodiment of the present invention, there is provided a kit comprising the HPMC powder composition defined herein and a signaling agent. Such kits can be used to treat influenza viruses, such as influenza A, H1N1, H5N1, and H3N2, coronaviruses, such as MERS-CoV, SAR-CoV, HCoV-229E, HCoV-NL63, HCoV It is intended to be used prophylactically or therapeutically to protect against viral attack from viruses such as CoV-OC43, CoV-HKU1, and 2019nCoV (also known as SAR-COV-2), as well as bacteria such as Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Haemophilus influenzae, Mycobacterium tuberculosis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecium, Candida albicans, Candida tropicalis, and Enterobacter species.

[0043] As suggested herein, the signaling agent additive may be selected from menthol, mint, spearmint, peppermint, eucalyptus, lavender, citrus, strawberry, or any combination thereof. Preferably, the signaling agent is one that is not known or implicated as an irritant, such as one selected from strawberry, lemon, lime, kumquat, or other citrus sources.

[0044] The signaling agent comprises from 0.25% to 10% or less, preferably from 0.50% to 5%, of the total weight of the applied composition.

[0045] According to a fourth embodiment of the present invention, there is provided a kit comprising an HPMC powder composition, a signaling agent, and a biologically active agent for simultaneous or sequential administration, wherein the signaling agent comprises 0.25% to 10% or less, preferably 0.50% to 5%, of the total weight of the composition. Such kits also protect against viral attacks from viruses such as influenza A viruses, e.g., H1N1, H5N1 and H3N2, coronaviruses, e.g., MERS-CoV, SAR-CoV, HCoV-229E, HCoV-NL63, HCoV-OC43, CoV-HKU1, and 2019-nCoV (also known as SAR-COV-2), as well as bacteria, e.g., Staphylococcus aureus, methicillin resistant Staphylococcus aureus, Haemophilus influenzae, Mycobacterium tuberculosis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, and others. baumannii, Enterococcus faecium, Candida albicans, Candida tropicalis, Enterobacter species, etc.

[0046] Suitable biologically active agents may be selected from St. John's wort, valerian extract, ginkgo biloba extract, vitamins A, E, or C, garlic, one or more probiotics, ginger, ellagic acid, echinacea, Swedish flower pollen, black walnut shells, lemongrass, mugwort, grapefruit seed extract, broccoli, digestive enzymes, hyaluronic acid, astragalus, rose hips, gentian, hypericum, horse chestnut, ginseng, green tea, phosphatidylserine, phosphatidylcholine, pycnogenol, caffeine, quercetin, coenzyme Q10, yarrow, tea tree, noni juice, lipase, fructooligosaccharides, inulin, black cumin, stabilized allicin, or any combination thereof.

[0047] Further suitable biologically active agents that show promise against diseases caused by coronaviruses, such as Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS), include type I (α, β) interferons (IFNs), such as IFN-β, IFNβ-1b, type II (γ) and type III (λ) interferons, remdesivir, ozeltamivir, zanamivir, ribavirin, lopinavir, the combination of lopinavir-ritonavir with IFNβ-1b, monoclonal and (camelid) polyclonal neutralizing antibodies and macrolides, such as ivermectin, plant alkaloids, such as colchicine. A compound that generally shows potential for use against coronaviruses is K22, structural name (Z)-N-(3-(4-(4-bromophenyl)-4-hydroxypiperidin-1-yl)-3-oxo-1-phenylprop-1-en-2-yl)benzamide, available from ChemDiv (San Diego, CA, catalog number 4295-0370). This compound targets membrane-bound viral RNA synthesis and shows potent inhibition in various coronaviruses, including MERS.Further suitable biologically active agents include the isolated griffithsin lectin protein of approximately 121 amino acids extractable from red algae such as Griffithsia, and its biologically active isolated antiviral analogs, and natural seaweed extracts containing the same (Journal of Virology 2010, O'Keefe, BR et al. "Broad Spectrum In Vitro Activity and In Vivo Efficacy of the Antiviral Protein Griffithsin against Emerging Viruses of the Family Coronaviridae." Online DOI Publication: 10.1128 / JVI.02322-09; Marine Drugs 2019 Oct.;17(10):567, Choongho Lee "Griffithsin - A Highly Potent Broad Spectrum Antiviral Lectin from Red Algae: From Discovery to Clinical Application"). "Lectin from Red Algae: From Discovery to Clinical Application," published online October 6, 2019, doi:10.3390 / md17100567.

[0048] Again, the signaling agent or additive may be selected from menthol, strawberry, mint, spearmint, peppermint, eucalyptus, lavender, citrus fruits, or any combination thereof. Preferably, the signaling agent is one that is not known or implicated as an irritant, such as lemon, lime, kumquat, or other citrus source.

[0049] The signaling agent comprises from 0.25% to 10% or less, preferably from 0.50% to 3%, of the total weight of the applied composition.

[0050] The powder composition of the present invention does not contain other additives or molecular components, as such additives may interfere with the ability of the composition of the present invention to form a gel when applied to the nasal mucosa. Such additives that are harmful to the formation of a gel in the nasal cavity include citric acid in combination with sodium citrate and benzalkonium chloride. Furthermore, other additives or components often used in intranasal compositions, such as other dry powders or solutions, may cause irritation or affect ciliary motility, for example, solvents such as propylene glycol, absorption enhancers such as cyclodextrins or glycosides, or mucoadhesives such as chitosan. The use of such additives may be undesirable because they may cause discomfort and interfere with normal nasal function, which may adversely affect breathing.

[0051] The powdered ingredients can be blended together for approximately 15 to 20 minutes using a ribbon blender or similar type blender. The blending time depends on the moisture content and compatibility of the powders. Preferably, the ingredients have a moisture content of less than 5% immediately after blending, as determined by the United States Pharmacopeia (USP / NF) Loss on Drying method.

[0052] Suitable devices for dispensing compositions according to the present invention are disclosed, for example, in European Patent Nos. 1,368,090 and 3,183,022, the teachings of which are incorporated herein in their entirety. The bottles disclosed therein use a very simple mechanism for limiting the amount of powder dispensed. While it is not necessary to precisely control the amount of powdered cellulose delivered to the nasal cavity to enhance natural mucus, administering too much powder can potentially cause uncomfortable blockage of the nasal cavity and even make breathing through the nose difficult.

[0053] Compositions according to the present invention are preferably administered in an amount of about 1 mg to about 10 mg per nostril. Preferably, the dose is about 2.5 mg to about 7.5 mg, 3 mg to about 7 mg, about 4 mg to about 6 mg, or about 5 mg.

[0054] In a fifth embodiment of the present invention, dry, free-flowing pHPMC particles for use in the present invention (i.e., before the addition of other dry powder components forming the compositions of the present invention) are provided, having the irregular sizes and shapes described herein and an average particle size of 118 μm when freshly prepared. Under storage conditions, moisture from the air is absorbed by the particles, causing them to swell by up to 14%, while still remaining in free-flowing powder form. Stored particles may assume an average particle size of up to about 134 μm. Thus, the HPMC particles themselves may have an average particle size of about 110 μm to 140 μm, preferably about 115 μm to about 135 μm, and more preferably 118 μm to 134 μm, depending on moisture absorption.

[0055] In a sixth embodiment of the present invention, there is provided a composition in homogenized dry powder form comprising hydroxypropyl methylcellulose particles and at least one chemical agent in particulate form selected from signal transduction agents, the homogenized powder comprising particles having an average particle size of 20 μm or more and 500 μm or less for use in the treatment or containment of respiratory disease in a mammal such as a human, the disease being caused by airborne allergens such as viruses, plant pollens and house dust mites, and / or PM 2.5 and PM 10 Such compositions may be used to treat respiratory illnesses caused by viruses such as coronaviruses selected from MERS-CoV, SAR-CoV, and 2019 nCov (also known as SAR-COV-2).

[0056] The composition of this embodiment of the invention preferably comprises powdered HPMC particles having an average particle size ranging from 60 to 150 μm, preferably 80 to 125 μm, more preferably 86 μm + / - 15 μm, and the signaling agent is selected from the group of mint, spearmint, peppermint, eucalyptus, lavender, citrus, or any combination thereof. Preferably, the signaling agent is selected from citrus, lemon, lime, kumquat, or any combination thereof. The signaling agent comprises no more than 0.25% to 10% of the total weight of the composition.

[0057] In a further embodiment of the present invention, there is provided a composition in the form of a homogenized dry powder comprising hydroxypropyl methylcellulose particles, at least one chemical agent in particulate form selected from signal transduction agents, and one or more biologically active agents, the homogenized powder comprising particles having an average particle size of between 20 μm and 500 μm, preferably between 60 μm and 150 μm, more preferably between 80 μm and 125 μm, for example 86 μm + / - 15 μm, for use as a medicament for respiratory disease in mammals such as humans, the disease being caused by airborne allergens such as viruses, plant pollens and house dust mites, and / or PM 2.5 and PM 10 Such compositions may be used to treat respiratory illnesses caused by viruses such as coronaviruses selected from MERS-CoV, SAR-CoV, and 2019-nCov (also known as SAR-COV-2). Definitions of all homogenized dry powder ingredients and amounts thereof in this aspect of the invention are as defined hereinabove.

[0058] In a further embodiment of the present invention, there is provided a composition as defined herein, which is for use as a nasally administered pharmaceutical agent.

[0059] As a still further embodiment of the present invention, there is provided a composition as defined herein, wherein the composition is for use in the treatment of COVID-19 disease.

[0060] As a still further embodiment of the present invention, there is provided a composition as defined herein, wherein the composition is for use in the prevention of COVID-19 disease.

[0061] Of course, those skilled in the art will understand that all compositional embodiments of the present invention detailed herein are for delivery to the nasal mucosa by insufflation through the nose.

[0062] 1) adding a signaling agent powder to a hydroxypropyl methylcellulose powder; 2) diffusively blending the two components of 1) in a blender; 3) optionally adding a powdered biologically active agent and further blending; Also provided is a method for producing a powder composition for use as a medicament for treating COVID-19 disease, comprising:

[0063] Examples and figures illustrating the invention are given below. It should be understood that the teachings of the examples and figures should not be construed as limiting the invention in any way. [Brief explanation of the drawings]

[0064] [Figure 1] Figure 1 shows the cumulative mass (average) of successive powder batches over a two year period, which were analyzed by particle size analyzer (Beckman Coulter LS particle size analyzer). [Figure 2] This figure shows the morphology of particles constituting HPMC as an important factor for deposition in the respiratory tract under a scanning electron microscope (magnification 100x). [Figure 3]FIG. 1 shows a step-by-step procedure for sample preparation, testing of pHPMC of the present invention and comparison with a competitor's hypromellose (cHPMC) powder against Der p 1 allergen using the agar diffusion method (in vitro). DETAILED DESCRIPTION OF THE INVENTION

[0065] Referring to the protocol in Figure 3, *For control samples: Follow the same procedure as above without adding extra thick sample layer and agar block. #For reference samples: follow the same procedure as above without adding the HPMC or hypromellose layer and Der p 1.

[0066] For the baseline measurement, 20 μl of Der p 1 antigen is added to 0.5 ml of PBS-T solution and the last three steps above are followed, followed by the ELISA measurement.

[0067] Experimental Section Section 1 Physicochemical characterization of powdered HPMC (HPMC) in support of its safety profile HPMC itself has been thoroughly characterized. As an inert natural product, its physical and biochemical properties do not provide any basis for safety concerns. Its favorable safety profile has been supported by all clinical trials conducted to date, none of which have reported any serious or severe adverse events (Popov TA, Aberg N, Emberlin J, et al. "Methyl-cellulose powder for prevention and management of nasal symptoms." Expert review of respiratory medicine. Nov 2017;11(11):885-892). 5、7A single ex vivo study suggests that higher doses of cellulose powder may negatively affect the viability of nasal epithelium and its ciliary beat frequency (Zhou M, Zuo KJ, Xu ZF, et al., "Effect of Cellulose Powder on Human Nasal Epithelial Cell Activity and Ciliary Beat Frequency." International Archives of Allergy and Immunology. 2019;178(3):229-237). 9 Nevertheless, because it is intended for use by nasal insufflation, we have thoroughly characterized the compound and performed toxicology studies in rats.

[0068] We routinely assayed HPMC batches by laser diffraction techniques to obtain the mean particle size. The particle size distribution was measured, with 99.4% of the particles falling within the diameter range of 5 to 500 μm, and the mean particle size being 118 μm (Figure 1).

[0069] Particle counts and mass distributions were measured in triplicate using a Grimm 1.109 laser particle counter connected to Grimm Dust Monitor 3.20 software. Test-retest reliability was assessed using correlation analysis, with Pearson coefficients of 0.998 and 0.985. The actual particle mass and count distributions proved variable, with a mean of 6,095.0 μg / m for all particle sizes. 3 The standard deviation of the mass distribution is 4,709.9 μg / m 3 (75.4% of the mean), and the mean of the count distribution is 619,135,967 counts / m 3 and the standard deviation is 330,964,124 counts / m 3(57.5% of the average). The particle size distribution of HPMC is highly skewed toward larger particles. The distribution pattern of HPMC particles depends on the actual delivery method utilized to deliver the powder, their morphology and swelling behavior due to the hygroscopic nature of cellulose. (Telko MJ, Hickey AJ. "Dry powder inhaler formulation." Respiratory care. Sep 2005;50(9):1209-1227) 10 HPMC particles are characterized by irregular shapes and surfaces that may affect nasal deposition (Figure 2). The coarse particle structure improves swelling by increasing the contact area, resulting in more efficient and faster swelling of the nose (Diethart B, "The use of inert hydroxypropyl methylcellulose powder as a remedy for allergic rhinitis. [Chapter 10, "The effect of HPMC application on human nasal cells"]: University of Coventry in collaboration with the University of Worcester; 2009).

[0070] Other determinants of nasal deposition include shape, density, potential charge, individual breathing pattern, and airflow velocity. Particles larger than 5 μm are deposited in the nasopharynx, while particles between 1 and 5 μm in diameter can deposit on the walls of the trachea and bronchial tree when actively inhaled. Particles deposited in the nose and tracheobronchial airways are trapped by the mucus layer, move with it to the pharynx, and are swallowed. Only particles smaller than 1 micron can reach the alveoli. In our study, only 0.63% of particles were smaller than 5 μm in diameter, and no particles smaller than 1.9 μm were detected. In other words, essentially none of the HPMC particles reached the alveoli, and therefore, it can be assumed that the entire amount was swallowed. This particle size range is advantageous for targeted deposition in the nasal cavity, achieving maximum local effectiveness in protecting the mucosa from allergens in allergic rhinitis and any irritants or infectious agents in non-allergic rhinitis.

[0071] Particle swelling begins immediately upon contact with moisture in the nasal cavity, and the powder also absorbs moisture from the nasal air, causing diameter growth. This is thought to result in increased intranasal deposition, the efficiency of which increases with increasing particle size. These inherent properties provide an explanation for the role HPMC may play in rapidly relieving the symptoms of seasonal allergic rhinitis. Overall, HPMC is a very safe material when administered orally in gram quantities, and the use of milligram quantities of Nasaleze for nasal insufflation does not pose any discernible risk. Based on a no-observed-adverse-effect level (NOAEL) of 5000 mg / kg bw / day from a 90-day feeding study in rats, a tolerable intake of 5 mg / kg bw / day for human consumption of HPMC was established, which is more than 100 times higher than the estimated current consumption of 0.047 mg / kg bw / day. (Burdock GA, "Safety assessment of hydroxypropyl methylcellulose as a food ingredient." Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association. December 2007; 45(12):2341-2351) 11 No genotoxicity or reproductive toxicity studies were identified, but the chemical properties of the materials, their recognized safety in food use, and lack of toxicity in feeding studies do not suggest that further research is needed.

[0072] In conclusion, in vitro studies support the ability of HPMC to form a gel upon contact with moisture, providing a reliable barrier against airborne allergens and particulate matter. Studies in rats also show that inhaling fairly high doses of HPMC via the mouth does not affect the animals' lungs, heart, or liver. In clinical practice, HPMC is not expected to be inhaled into the lower respiratory tract, and the cited animal studies provide an additional precaution: no adverse consequences are expected even if this unintentional event occurs.

[0073] Expert commentary Preventing contact between the nasal mucosa and harmful substances in the surrounding environment (allergens, irritants, microorganisms) that attack it is the simplest and most natural approach to prevent inflammatory events in the airways and the subsequent induction of clinical symptoms. This approach is called "barrier-forcing measures" and can be considered a means to achieve allergen avoidance (Andersson M, Greiff L, Ojeda P, Wollmer P. "Barrier-forcing measures as a treatment principle in allergic rhinitis: a systematic review." Current medical research and opinion. Jun 2014;30(6):1131-1137). 12 .

[0074] Ideally, if properly implemented, this strategy could obviate the need for any other therapeutic agents. Attempts have been made to use various substances as barrier enhancers, namely white petrolatum, pollen blocker creams, lipid-based ointments, microemulsions, liposomal preparations, and seawater gels.

[0075] Many of the approaches listed have not stood the test of time and have been abandoned. Microcrystalline powder hydroxypropyl methylcellulose (HPMC) has been developed into a patented medical device and licensed for the management of allergic rhinitis (general product information is available at https: / / www.nasaleze.com / ). Its clinical and real-world efficacy has been proven in dozens of studies. There are unresolved issues along the way, which have been considered and tested in laboratory, in vitro, and ex vivo studies. This overview provides previously unpublished data that may be useful to the medical and patient communities as a basis for the broader application of natural products for the prevention and treatment of airway diseases.

[0076] Key Issues HPMC is a cellulose derivative powder with a patented drug delivery system. Intranasally insufflated HPMC emits a spectrum of particles, 99.4% of which are within the diameter range of 5 to 500 μm. HPMC particles are highly hygroscopic and have a rough shape and surface, resulting in rapid swelling and gel formation when insufflated into the nose. The HPMC gel layer acts as a barrier between the nasal mucosa and pollen, dust mite allergens, and 2.5 μm particulate matter (PM 2.5 ) to prevent contact with them (avoidance effect). In addition to theoretical arguments and long-term experience with cellulose derivatives, studies in rats have demonstrated that HPMC does not deposit in the lungs and does not cause adverse systemic effects.

[0077] Section 2 Description of particle size before and after homogeneous mixing of cellulose with signaling agents Particle size definition of IOM915K HPMC cellulose powder Pure HPMC, designated IOM915K, is a polydisperse powder that specifically targets the extrathoracic airways. When instilled intranasally, over 96% of the IOM915K powder is available for immediate gel formation.

[0078] Initial particle size measurements showed that the pure cellulose powder varied between 2 and 478.50 microns, with a mean particle size of approximately 118 μm. The introduction of signaling agents designed to allow the end user to determine when an effective dose had been injected was introduced in 2006 after reports of difficulties in ensuring that the dose had been injected in early clinical trial studies (Josling P, Steadman S, "Use of cellulose powder for the treatment of seasonal allergic rhinitis," Adv Therapy 20, 213-219, 2003). 13 and Emberlin JC and Lewis RA, "A double blind placebo controlled trial of inert cellulose for the relief of hay fever in adults," Current Med Research and Opinion, 22, 275-285, 2006. 14 .

[0079] This means that a powder homogenization and controlled mixing procedure was adopted as follows.

[0080] Powder homogenization We use IOM915K HPMC powder and mix it with our established signaling agents including lemon, mint, garlic and strawberry under standard operating procedures.

[0081] QMS Procedure 4, Revision 11, dated July 10, 2018. Powders are routinely assayed for moisture content, density, and particle size, as well as standard microbiological analysis. Powder blending is completed using a V-blender for a total of 15 minutes for each blend. This follows our protocol for powder storage, blend preparation, blend proportion calculation, and use of an industrial-grade V-blender machine (V100, model number A39525-2, supplied by Key Packaging Machinery Limited).

[0082] A V-blender consists of two hollow cylindrical shells joined at a 75° to 90° angle. The blender vessel is mounted on a trunnion to allow rotation. As the V-blender rotates, the material is continuously divided and recombined, and mixing occurs as the material falls freely into the vessel. The repeated converging and diverging motion of the material, combined with increased frictional contact between the material and the long, straight sides of the vessel, results in a gentle but uniform mixture.

[0083] The primary mechanism of blending in a V-blender is diffusion. Diffusion blending is characterized by small-scale random motion of solid particles. The motion of the blender increases the mobility of individual particles, thus promoting diffusive blending. Diffusive blending occurs when particles are distributed over newly developed interfaces. In the absence of segregation effects, diffusion blending eventually leads to a high degree of homogenization. Therefore, V-blenders are preferred when precise blend formulations are required. V-blenders are also well suited for applications where some components may be as low as 5 percent of the overall blend size, as in the homogenization between IOM915K and a signaling agent that is present at less than 5% of the overall blend mixture. Typical blending times of 15 minutes ensure complete homogenization of the powders of the present invention.

[0084] IOM915K+ Signaling Agent Particle Size Definition After homogenization of the powder, the particle size changes significantly from pure IOM915K. Upon mixing, a proportion of the larger particles are reduced in size due to being broken down in the V-blender when mixed with the signaling agent. Similarly, a small proportion of the smaller particles, less than 5 microns, aggregate to form larger particles. Analysis of particle size shows that the powder mixture ranges from 4 to 395 microns, with an average of 86.2 μm.

[0085] Further studies also show that over time, ie, over a period of several months, the overall particle size average tends to increase.

[0086] HPMC mixtures increase in size by approximately 14% during storage at ambient temperature. Therefore, we hypothesize that our powders absorb moisture from the air, increasing in diameter and causing the powder to deposit higher in the respiratory tract upon nasal instillation. This may result in increased nasal deposition, with increasing efficiency with increasing particle size.

[0087] From the extensive clinical trial database now available with our Nasaleze family of extracts containing IOM915K cellulose and proprietary formulations of lemon, mint, strawberry and garlic, we have demonstrated that small particles do not reach the lungs or brain, and that our mean particle size of 86.2 μm, along with the associated mass of these particles, provides highly effective control of symptoms in persistent allergic rhinitis, as well as protection from pollen, viruses, bacteria, fungi and PM 2.5 and PM 10 It has been shown that this technology enables the elimination of pathogens, including environmental toxins such as

[0088] Section 3 Section 3(a) Determination of the preventive and therapeutic potential of pHPMC powder formulated with mint and wild garlic extracts as signaling agents against coronavirus 229E.

[0089] 1.0 Purpose To determine the antiviral efficacy of Nasaleze® powder (mean particle size approximately 82 μm) against human coronavirus 229E (CoV229E). 5% w / w European wild garlic extract was obtained from Pfannenschmidt GmbH, Hamburg, Germany. 93% w / w Nasaleze® powder from Nasaleze Limited (locally)

[0090] 2.0 Materials and Methods 2.1 Test organisms Cell type: Medical Research Council human fibroblast cell line 5 [MRC-5 (ATCC® CCL-171)] Virus: Human coronavirus 229E (CoV229E) (ATCC® VR-740)

[0091] 2.2 Test Agents The test agents used in this study are shown in Table 1.

[0092] [Table 1]

[0093] 2.3 Devices and Media device: Class II Biosafety Cabinet - BioMAT, ThermoFisher Scientific, UK Vortex - Grant Instruments, UK UKAS Calibrated Multichannel Pipette (P300) - Gilson®, UK UKAS Calibrated Multichannel Pipette (P20) - Gilson®, UK UKAS calibrated pipettes (0.5-1000 μL range) - Proline® Plus, UK 96-well plates - ThermoFisher Scientific, UK CO2 Incubator - Thermo Scientific, UK Tissue culture flasks - Nunc, ThermoFisher Scientific, UK Olympus CK2 inverted microscope - KeyMed, UK VWB2 Bather - VWR, UK Vacuboy Aspirator-INTEGRA, UK Medium: Phosphate Buffered Saline (PBS) - Gibco™, UK Penicillin-Streptomycin - ThermoFisher Scientific, UK, Eagle's Minimum Essential Medium (EMEM) - ATCC®, UK Dulbecco's Phosphate Buffered Saline (DPBS) Gibco(TM), UK Fetal Bovine Serum (FBS) - Gibco™, USA Trypsin-EDTA-Gibco™, UK Trypan blue - Sigma-Aldrich, UK

[0094] 2.4 Method 2.4.1 Cell maintenance and assay setup MRC-5 cells were used as a host cell line for human coronavirus 229E (CoV229E) propagation. MRC-5 cells were maintained in Eagle's minimum essential medium (EMEM) supplemented with 20% fetal bovine serum (FBS) and 1% penicillin-streptomycin (complete EMEM) at 37 ± 2°C and 5% CO2. In preparation for cytotoxicity screening and antiviral assays, MRC-5 cells were plated in 24-well plates at 1.0 × 10 5 MRC-5 cells were seeded at 2 × 10 cells / mL and incubated at 37 ± 2°C and 5% CO2 for 24 hours or until they reached 80–90% confluency. In preparation for tissue culture infectious dose 50 (TCID50) testing, MRC-5 cells were seeded at 2 × 10 cells / mL in 96-well plates. 5 cells mL -1 and incubated at 37±2°C and 5% CO2 for 24 hours.

[0095] 2.4.2 Phase 1: Cytotoxicity Screening of Nasal Spray Formulations Nasaleze® HPMC powder was diluted to 3.2 mg / 0.1 mL, 6.4 mg / 0.1 mL, and 12.8 mg / 0.1 mL in EMEM supplemented with 2% FBS and 1% penicillin-streptomycin (assay medium). Complete EMEM was aspirated from the test plate, and 100 μL of each test concentration was added to replicate wells. After a 10-minute incubation period at 20 ± 2°C, an additional 400 μL of assay medium was added to the test wells. The plates were incubated at 37 ± 2°C and 5% CO2 for 24 hours. After incubation, visual scoring was performed according to ISO 10993-5 guidelines on a scale of 0 to 4 (Table 2). Cytotoxic effects were assessed based on various morphological changes to MRC-5 cells, such as cell rounding, detachment, and cell lysis.

[0096] [Table 2]

[0097] 2.4.3 Phase 2: Evaluation of the preventive and therapeutic potential of Nasaleze® Powder MRC-5 cells were treated with Nasaleze® powder according to two methods to determine the preventative and therapeutic capabilities of the formulation. Assays were performed in 24-well plates with replicate wells for each experimental condition.

[0098] 2.4.3.1 Prophylactic Treatment of MRC-5 Cells with Nasaleze® Powder Prior to Human Coronavirus 229E Infection To evaluate the protective potential of Nasaleze® powder against CoV229E, MRC-5 cells were pretreated with 3.2 mg of the formulation for 10 minutes and then infected with CoV229E at a multiplicity of infection (MOI) of 1 (high dose) and 0.01 (low dose). After aspirating complete EMEM from the test plates and washing once with Dulbecco's phosphate-buffered saline (DPBS), 3.2 mg of Nasaleze® powder was applied in 100 μL of assay medium. After 10 minutes of incubation at 20 ± 2°C, cells were inoculated with 100 μL of CoV229E, pre-diluted to achieve high and low MOI infection, and incubated for 30 minutes at 35 ± 2°C and 5% CO2. The infected cells were then supplemented with an additional 300 μL of assay medium and incubated at 35 ± 2°C and 5% CO2 for 4 days. Viral cytopathic effect (CPE) on MRC-5 cells was scored on days 2, 3, and 4 according to the criteria listed in Table 2. On days 3 and 4, 100 μL of medium was collected from each well to determine the viral titer and then replaced with 100 μL of fresh assay medium. Collected samples were stored at -80°C until required for viral titer determination.

[0099] 2.4.3.2 Treatment of Human Coronavirus 229E-Infected MRC-5 Cells with Nasaleze® Powder To evaluate the therapeutic potential of Nasaleze® powder against CoV229E, MRC-5 cells were first infected with CoV229E at a high MOI of 1 and a low MOI of 0.01, respectively, and then treated with the formulation. After aspirating complete EMEM from the test plates and washing once with DPBS, 100 μL of pre-diluted CoV229E was inoculated to achieve high and low MOI infections and incubated for 30 minutes at 35 ± 2°C and 5% CO2. After incubation, the virus inoculum was removed, and a 3.2 mg dose of Nasaleze® powder in 100 μL assay medium was added to the cells and incubated for 10 minutes at 20 ± 2°C to allow gel barrier formation. The treated cells were then supplemented with an additional 300 μL of assay medium and incubated for 4 days at 35 ± 2°C and 5% CO2. Viral CPE on MRC-5 cells was scored on days 2, 3, and 4 using the criteria described in Table 2. On days 3 and 4, 100 μL of medium was collected from each well to determine the virus titer and then replaced with another 100 μL of fresh assay medium. The collected samples were stored at -80°C until required for virus titration.

[0100] 2.4.4 TCID 50 Quantification of viral infectivity by To determine the viral titer of the collected samples, 10-fold serial dilutions were performed in assay medium. The medium was aspirated from the wells of the cell plate, and the cells were washed with DPBS. 100 microliters of each dilution of sample was added to the corresponding test well. The test plate was incubated at 35±2°C and 5% CO2 for 7 days. There were four replicate wells for each test condition. After incubation, viral CPE was determined using an Olympus CK2 inverted microscope. The viral titer was calculated using the Spearman-Carber method.

[0101] 3.0 Results 3.1 Phase 1: Cytotoxicity Screening No observable cytotoxicity was observed in MRC-5 cells exposed to Nasaleze® powder after a 24-hour contact period (Table 3). Visual scoring revealed a gel barrier formed by the Nasaleze® powder above the cell monolayer. Additionally, residue was observed on the treated cells (data not shown).

[0102] [Table 3]

[0103] 3.2 Prophylactic Treatment of MRC-5 Cells with Nasaleze® Powder Prior to Coronavirus 229E Infection 3.2.1. Cytopathic Effect of CoV229E on MRC-5 Cells Pretreated with Nasaleze® Powder After incubation periods of 2, 3, and 4 days, the test plates were scored for CPE (Tables 4-6). CPE was observed (visual data not shown). Replicating cells treated with a high MOI of CoV229E and Nasaleze® powder showed slight CPE on day 2 and severe CPE on days 3 and 4. Replicating cells treated with a low MOI of CoV229E and Nasaleze® powder showed no CPE on day 2 and moderate CPE on days 3 and 4.

[0104] [Table 4]

[0105] [Table 5]

[0106] [Table 6]

[0107] 3.2.2 Viral Titration of Samples Pretreated with Nasaleze® Powder After 3-day and 4-day incubation periods with high MOI CoV229E, the negative control resulted in mean virus titers of 5.82±0.35 Log10TCID50 / mL and 5.32±0.35 Log10TCID50 / mL, respectively. Pretreatment of MRC-5 cells with Nasaleze® powder resulted in a reduction in virus titer of 2.68 Log10TCID50 / mL and 2.55 Log10TCID50 / mL on days 3 and 4 post-infection, respectively, when compared to the negative control (Table 7).

[0108] [Table 7]

[0109] After 3-day and 4-day incubation periods with low MOI CoV229E, the negative control resulted in mean virus titers of 6.02±0.53 Log10TCID50 / mL and 5.39±0.18 Log10TCID50 / mL, respectively. Pretreatment of MRC-5 cells with Nasaleze® powder resulted in a reduction in virus titer of 1.70 Log10TCID50 / mL and 1.00 Log10TCID50 / mL on days 3 and 4 post-infection, respectively, when compared to the negative control (Table 8).

[0110] [Table 8]

[0111] 3.3 Therapeutic Potential of Nasaleze® Powder 3.3.1 Cytopathic Effect of CoV229E on MRC-5 Cells Treated with Nasaleze® Powder After Viral Infection After incubation periods of 2, 3, and 4 days, the test plates were scored for CPE (Tables 9-11). Representative images of observed CPE are shown in Figure B. Replicating cells infected with a high MOI of CoV229E and treated with Nasaleze® powder showed mild CPE on day 2 and severe CPE on days 3 and 4 post-infection. Replicating cells infected with a low MOI of CoV229E and treated with Nasaleze® powder showed no CPE on day 2 and moderate CPE on days 3 and 4 post-infection.

[0112] [Table 9]

[0113] [Table 10]

[0114] [Table 11]

[0115] 3.3.2 Viral Titer Measurements in Samples Treated with Nasaleze® Powder After Viral Infection After 3-day and 4-day incubation periods with high MOI CoV229E, the negative control resulted in mean virus titers of 5.82±0.35 Log10TCID50 / mL and 5.32±0.35 Log10TCID50 / mL, respectively. Treatment of MRC-5 cells with Nasaleze® powder following infection with high MOI CoV229E resulted in a reduction in virus titer of 1.07 Log10TCID50 / mL and 1.93 Log10TCID50 / mL on days 3 and 4 post-infection, respectively, when compared to the negative control (Table 12).

[0116] [Table 12]

[0117] After 3-day and 4-day incubation periods with low MOI CoV229E, the negative control resulted in mean virus titers of 6.50±0.00 Log10TCID50 / mL and 5.89±0.18 Log10TCID50 / mL, respectively. Treatment of MRC-5 cells with Nasaleze® powder following infection with low MOI CoV229E resulted in a reduction in virus titer of 0.75 Log10TCID50 / mL and 1.00 Log10TCID50 / mL on days 3 and 4 post-infection, respectively, when compared to the negative control (Table 13).

[0118] [Table 13]

[0119] 4.0 Discussion The spread of potentially pathogenic viruses increases the risk of infection in both healthy and immunocompromised individuals. Coronaviruses are enveloped, single-stranded RNA viruses that cause a variety of upper respiratory tract illnesses in humans. These illnesses range from mild symptoms, such as the common cold, to severe acute respiratory syndromes, as seen in the recent COVID-19 pandemic. Coronaviruses are thought to be transmitted primarily via respiratory droplets, and some evidence suggests that the virus can remain active on fomites for several days. Both preventive and therapeutic interventions are essential to slow and / or stop the spread of coronaviruses. Evaluation of this invention against coronavirus surrogate strains allows for the safe evaluation of product efficacy. Coronavirus 229E is structurally and genetically similar to the SARS-CoV-2 virus.

[0120] Two approaches were used to investigate the antiviral effects of Nasaleze® powder. In the first group of studies, MRC-5 cells were pretreated with Nasaleze® powder and then infected with high and low doses of CoV229E. In the second group, MRC-5 cells were infected with high and low doses of CoV229E before treatment with Nasaleze® powder. Treatment with Nasaleze® powder resulted in a substantial reduction in viral titer in both experimental groups, demonstrating high levels of antiviral activity.

[0121] Section 3(b) 1.0 Purpose To evaluate the antiviral efficacy of two nasal dry powder spray products against human coronavirus 229E using a preventative and treatment-based approach.

[0122] 2.0 Materials and Methods 2.1 Test organisms Cell type: MRC-5 (ATCC® CCL-171™) passage 3 Virus: Human coronavirus 229E (CoV229E) (ATCC® VR-740™) - Amplification Number: 1

[0123] 2.2 Test Agents The test agents used in the study are listed in Table 1.

[0124] [Table 14]

[0125] 1. REM consists of remdesivir at a concentration of 8% w / w homogeneously mixed with 90% HPMC particles and 2% signaling agent.

[0126] 2. IVER consists of ivermectin at a concentration of 8% w / w homogeneously mixed with 90% w / w HPMC particles and 2% signaling agent.

[0127] 2.3 Devices and Media device: Class II Biosafety Cabinet - BioMAT, ThermoFisher Scientific, UK Vortex - Grant Instruments, UK UKAS Calibrated Multichannel Pipette (P300) - Gilson®, UK UKAS Calibrated Multichannel Pipette (P20) - Gilson®, UK UKAS calibrated pipettes (0.5-1000 μL range) - Proline® Plus, UK 96-well plates - ThermoFisher Scientific, UK 24-well plate - ThermoFisher Scientific, UK CO2 Incubator BB-15 - Thermo Scientific, UK Tissue culture flasks - Nunc, ThermoFisher Scientific, UK Olympus CK2 inverted microscope - KeyMed, UK VWB2 Water Bath - VWR, UK Vacuboy Aspirator INTEGRA, UK Medium: Phosphate Buffered Saline (PBS) - Gibco™, UK Penicillin-Streptomycin - ThermoFisher Scientific, UK Eagle's Minimum Essential Medium (EMEM) ATCC®, UK Dulbecco's Phosphate Buffered Saline (DPBS) Gibco™, UK Fetal Bovine Serum (FBS) - Gibco™, USA Trypsin-EDTA Gibco™, UK Trypan Blue Sigma Aldrich, UK

[0128] 2.4 Method 2.4.1 Cell maintenance and assay setup MRC-5 cells were used as a host cell line for propagation of human coronavirus 229E. MRC-5 cells were maintained in Eagle's minimum essential medium (EMEM) supplemented with 20% fetal bovine serum (FBS) and 1% penicillin-streptomycin (complete EMEM) at 37 ± 2 °C and 5% CO2. In preparation for cytotoxicity screening and antiviral assays, MRC-5 cells were seeded into 24-well plates and incubated at 37 ± 2 °C and 5% CO2 for 24 h or until they reached 80-90% confluence.

[0129] 2.4.2 Phase 1: Cytotoxicity Screening of Nasal Spray Formulations Test articles were diluted to 3.2 mg / 0.1 mL in EMEM supplemented with 2% FBS and 1% penicillin-streptomycin (assay medium). Complete culture EMEM was aspirated from the test plate, and 100 μL of each test concentration was added to replicate wells. After a 10-minute incubation period at 20 ± 2°C, an additional 400 μL of assay medium was added to the test wells. The plates were incubated at 37 ± 2°C and 5% CO2 for 24 hours. After incubation, visual scoring was performed according to ISO 10993-5 guidelines on a scale of 0 to 4 (Table 2). Cytotoxic effects were assessed based on various morphological changes to MRC-5 cells, such as cell rounding, detachment, and cell lysis.

[0130] [Table 15]

[0131] 2.4.3 Phase 2: Antiviral Efficacy of Two Nasal Spray Formulations Against Human Coronavirus 229E Using Prevention-Based and Treatment-Based Approaches MRC-5 cells were treated with the nasal spray formulations according to two methods to determine the preventative and therapeutic potential of the formulations. Assays were performed in 24-well plates with replicate wells for each experimental condition.

[0132] 2.4.3.1 Prophylactic treatment of MRC-5 cells with two nasal spray formulations prior to infection with human coronavirus 229E To evaluate the protective potential of the nasal spray against human coronavirus 229E, MRC-5 cells were pretreated with 3.2 mg / 0.1 mL of each formulation for 10 minutes prior to infection. Complete EMEM was aspirated from the test plates, washed once with Dulbecco's phosphate-buffered saline (DPBS), and 3.2 mg of test powder in 100 μL of assay medium was applied. After incubation at 20 ± 2°C for 10 minutes, cells were inoculated with 100 μL.

[0133] 2.4.3.2 Prophylactic treatment of MRC-5 cells with two nasal spray formulations prior to infection with human coronavirus 229E To evaluate the protective potential of the nasal spray against human coronavirus 229E, MRC-5 cells were pretreated with 3.2 mg / 0.1 mL of each formulation for 10 minutes prior to infection. Complete EMEM was aspirated from the test plate, washed once with Dulbecco's phosphate-buffered saline (DPBS), and 3.2 mg of test powder in 100 μL of assay medium was applied. After a 10-minute incubation at 20 ± 2°C, the cells were inoculated with 100 μL of human coronavirus 229E, prediluted to achieve high (0.3) and low (0.01) multiplicities.

[0134] 2.4.3.3 Prophylactic treatment of MRC-5 cells with two nasal spray formulations prior to infection with human coronavirus 229E To evaluate the protective potential of the nasal spray against human coronavirus 229E, MRC-5 cells were pretreated with 3.2 mg / 0.1 mL of each formulation for 10 minutes prior to infection. Complete EMEM was aspirated from the test plate, washed once with Dulbecco's phosphate-buffered saline (DPBS), and 3.2 mg of test powder in 100 μL of assay medium was applied. After 10 minutes of incubation at 20 ± 2°C, cells were inoculated with 100 μL of human coronavirus 229E, pre-diluted to achieve high (0.3) and low (0.01) multiplicities of infection (MOI). Samples were incubated for 30 minutes at 35 ± 2°C and 5% CO2. The infected cells were then supplemented with an additional 300 μL of assay medium and incubated for 4 days at 35 ± 2°C and 5% CO2. On days 2, 3, and 4, 100 μL of medium was collected from each well to determine virus titers. A 100 μL aliquot of fresh assay medium was applied to the cells after each harvest. Harvested samples were stored at -80°C until required for viral titration by TCID50. Viral titers were calculated using the Spearman-Carber method.

[0135] 2.4.3.4 Treatment of human coronavirus 229E-infected MRC-5 cells with two nasal spray formulations To evaluate the therapeutic potential of the nasal spray against human coronavirus 229E, MRC-5 cells were first infected with human coronavirus 229E at a high MOI of 0.3 and a low MOI of 0.01, respectively, and then treated with each of these two formulations. Complete EMEM was aspirated from the test plates and washed once with DPBS. Samples were inoculated with 100 μL of pre-diluted human coronavirus 229E to achieve high and low MOI infections and incubated for 30 minutes at 35 ± 2°C and 5% CO2. After incubation, the virus inoculum was removed, and a 3.2 mg dose of test powder in 100 μL assay medium was added to the cells and incubated for 10 minutes at 20 ± 2°C to allow gel barrier formation. The treated cells were then supplemented with an additional 300 μL of assay medium and incubated for 4 days at 35 ± 2°C and 5% CO2. On days 2, 3, and 4, 100 μL of medium was collected from each well to determine the virus titer. A 100 μL aliquot of fresh assay medium was applied to the cells after each collection. Collected samples were stored at -80°C until required for virus titration by TCID50. Virus titers were calculated using the Spearman-Carber method.

[0136] 3.0 Results 3.1 Phase 1: Cytotoxicity Screening of Two Nasal Spray Formulations No observable cytotoxicity was observed in MRC-5 cells exposed to the nasal spray after a 24-hour contact period (Table 3).

[0137] [Table 16]

[0138] 3.2 Phase 2: Antiviral efficacy of two nasal spray formulations against human coronavirus 229E using prevention-based and treatment-based approaches. 3.2.1 Prophylactic treatment of MRC-5 cells with a nasal spray formulation prior to human coronavirus 229E infection. 3.2.1.1 High MOI After 2, 3, and 4 days of incubation with a high MOI of human coronavirus 229E, the positive infection control had 7.00, 6.50, and 4.75 Log10TCID50mL, respectively. -1 Pretreatment of MRC-5 cells with 1.REM and 2.IVER resulted in the highest reduction in human coronavirus 229E recovered after 2, 3, and 4 days of incubation (Table 4).

[0139] [Table 17]

[0140] 3.2.1.2 Low MOI After 2, 3, and 4 days of incubation with a low MOI of human coronavirus 229E, the positive infected control had 7.50, 6.67, and 6.42 Log10TCID50mL, respectively. -1 Pretreatment of MRC-5 cells with 1.REM and 2.IVER resulted in the highest reduction in human coronavirus 229E recovered after 2 and 4 days of incubation (Table 5).

[0141] [Table 18]

[0142] 3.2.2 Treatment of human coronavirus 229E-infected MRC-5 cells with two nasal spray formulations 3.2.2.1 High MOI After 2, 3, and 4 days of incubation with a high MOI of human coronavirus 229E, the positive infection control had 7.00, 6.50, and 4.75 Log10TCID50mL, respectively. -1Treatment of human coronavirus 229E-infected MRC-5 cells with 1.REM and 2.IVER resulted in the highest reduction in human coronavirus 229E recovered after 2, 3, and 4 days of incubation (Table 6).

[0143] [Table 19]

[0144] 3.2.2.2 Low MOI After 2, 3, and 4 days of incubation with a low MOI of human coronavirus 229E, the positive infected control had 7.50, 6.67, and 6.42 Log10TCID50mL, respectively. -1 Treatment of human coronavirus 229E-infected MRC-5 cells with 1.REM and 2.IVER resulted in the highest reduction in human coronavirus 229E recovered after 2, 3, and 4 days of incubation (Table 7).

[0145] [Table 20]

[0146] 4.0 Discussion The spread of potentially pathogenic viruses increases the risk of infection in both healthy and immunocompromised individuals. Coronaviruses are enveloped, single-stranded RNA viruses that cause a variety of upper respiratory tract diseases in humans. These diseases range from mild symptoms (e.g., the common cold) to severe acute respiratory syndromes, as seen in the ongoing COVID-19 pandemic. Interventions that take both preventive and curative approaches are essential to halt or slow the spread of coronaviruses. In this study, the preventive and curative applications of two formulations were evaluated against high and low doses of human coronavirus 229E. Coronavirus 229E is structurally and genetically similar to the SARS-CoV-2 virus.

[0147] Across all evaluations, REM and IVER resulted in a reduction in human coronavirus 229E recovered after prophylactic and curative applications.

[0148] Future studies could investigate the effects of the formulation after multiple applications. Future studies could also evaluate the nasal spray formulation against other respiratory viruses, such as influenza A and B, adenovirus, and rhinovirus. Bacterial respiratory pathogens, such as Pseudomonas aeruginosa, could also be investigated. To further mimic the actual use of the product, a 3D nasal model could be used to understand the effect on ciliary function after application of the formulation.

[0149] Section 4 Preamble In the art, hydroxypropyl methylcellulose (HPMC) is also known by the synonym "hypromellose." The term "hypromellose" is used in product literature for competing products. To distinguish the results of Applicant's HPMC-containing powders from those of competitors, "hypromellose" is used in Section 4 to distinguish them from Applicant's HPMC-containing powders.

[0150] It should be understood that all references herein to "hypromellose" refer only to competitive low pH hypromellose-containing compositions that further contain additives that act to lower their pH after contact with moisture. The HPMC-containing powders of the present invention do not contain additives of the type known to be included in competitive products.

[0151] One purpose of Section 4 is to compare the performance of the HPMC powder of the present invention with that of a competitor hypromellose-containing powder.

[0152] The competitive hypromellose-containing product to which Applicant's HPMC-containing powder is being compared has the following ingredients: 89.9% hypromellose, 6% citric acid, 4% sodium citrate, 0.1% benzalkonium chloride and less than 0.1% menthol as described in the literature for the competitive product.

[0153] Section 4 Application of hydroxypropyl methylcellulose gel retards Der p 1 diffusion in vitro significantly better than low pH hypromellose background: Following updated ARIA guidelines and data indicating that certain cellulose powders can trap viral particles by forming an internal gel barrier within the nose, we investigated hydroxypropyl methylcellulose powders of different average particle sizes and a competing, commercially available, low-pH hypromellose powder for the relief of nasal symptoms of allergic rhinitis and the capture of viral particles, including coronaviruses 229E and SARS-CoV-2. The efficacy of these barrier compounds has been the subject of several clinical, observational, and in vitro studies. The purpose of this study was to investigate the hypothesis that the quality of the gel formed after nasal moisture absorption may be related to the average particle size, with lower particle sizes producing a less effective barrier against external pathogens. The quality of the mechanical barrier created by each compound is also important in preventing allergen diffusion to the nasal epithelium over an extended period of time.

[0154] Methods: The diffusion of Der p 1 through HPMC and hypromellose gels was measured in vitro after 15, 30, 60, 180, and 360 minutes using an ELISA method. Agar blocks were used to simulate the nasal mucosa. Control samples without the gel layer were obtained.

[0155] Section 4 Application of hydroxypropyl methylcellulose gel retards Der p 1 diffusion in vitro significantly better than low pH hypromellose background: Following updated ARIA guidelines and data indicating that certain cellulose powders can trap viral particles by forming an internal gel barrier within the nose, we investigated hydroxypropyl methylcellulose powders of different average particle sizes and a competing, commercially available, low-pH hypromellose powder for the relief of nasal symptoms of allergic rhinitis and the capture of viral particles, including coronaviruses 229E and SARS-CoV-2. The efficacy of these barrier compounds has been the subject of several clinical, observational, and in vitro studies. The purpose of this study was to investigate the hypothesis that the quality of the gel formed after nasal moisture absorption may be related to the average particle size, with lower particle sizes producing a less effective barrier against external pathogens. The quality of the mechanical barrier created by each compound is also important in preventing allergen diffusion to the nasal epithelium over an extended period of time.

[0156] Methods: The diffusion of Der p 1 through HPMC and hypromellose gels was measured in vitro after 15, 30, 60, 180, and 360 minutes using an ELISA method. Agar blocks were used to simulate the nasal mucosa. Control samples without the gel layer were obtained.

[0157] result: The control sample, which did not have a gel barrier applied, absorbed 100% of the Der p 1 solution after 15 minutes. In comparison, HPMC significantly delayed Der p 1 diffusion, allowing only 1.33% penetration into the agar block after 15 minutes and only 10.41% after 360 minutes under simulated nasal conditions; slight differences were observed between small, medium, and larger particle sizes, all of which were superior to hypromellose gel, which allowed 5.37% penetration after 15 minutes and 25.89% penetration after 360 minutes under the same conditions.

[0158] Conclusion: HPMC gel significantly reduces Der p 1 diffusion in vitro compared to hypromellose, which may be due to the average mesh size of the polymer network of HPMC forming a more efficient barrier than the lower mesh size of hypromellose, and may have important implications for the formation of a prophylactic barrier to capture various pathogens.

[0159] method Three HPMC compounds were constructed and provided for testing by Nasaleze Limited. A sample of a low pH hypromellose compound was obtained from Nasus Pharma, IL.

[0160] Der p 1 allergen was procured from Indoor Biotechnology, India. The experiment was followed as per the stepwise protocol shown in Figure 3. ELISA measurements were then performed.

[0161] ELISA measurement The Der p1 allergen standard used in the assay was purchased from Indoor Biotechnologies and the assay was performed according to the manufacturer's instructions.

[0162] Results and findings The average baseline allergen content in 20 ul of standard solution was found to be 153.02 ng after recommended dilution and preparation of the stock solution.

[0163] The results and observations clearly show that all three HPMC powders were free flowing in nature when sprayed from the container (traditional powder spray bottle), whereas the hypromellose powder had to be tapped several times to become free flowing from the bottle.

[0164] All HPMC formulations immediately formed thick, clear, firm gels when mixed with the diluent, whereas hypromellose did not initially form any kind of gel, as it was actually a liquid. After a series of dilutions, all gels from the samples were confirmed to be in 5% gel solution by mixing 50 mg of powder with 1 ml of 0.9% sterile saline to match the pH and consistency of normal nasal mucosa.

[0165] Throughout the experiment, and even after 6 hours of incubation at 30–35°C, the HPMC layer remained thick and fresh in morphology, while the hypromellose layer dried completely and formed a white precipitate on the surface of the glass slide.

[0166] [Table 21] pHPMC small particles: average particle size = 88.57μm pHPMC medium particles: average particle size = 107.7μm pHPMC large particles: average particle size = 121.00μm Hypromellose (competitive low pH hypromellose): average particle size = 68.56 μm

[0167] conclusion The data clearly show that HPMC is superior to the competitor hypromellose in terms of the quality, consistency and nature of the barrier produced, and this means that HPMC is approximately 150% more effective than hypromellose in preventing penetration by Der p 1 allergen over the 360 ​​minute test; therefore, the inventors predict that when HPMC is used, the ability to trap allergens including pollen, viruses, bacteria and spores in the nasal mucosa will be much more efficient.

[0168] References TIFF2026042058000023.tif48169 TIFF2026042058000024.tif214169

Claims

1. 1. A composition in the form of a homogenized dry powder for use in the prevention and / or treatment of coronavirus diseases, said composition comprising: i) hydroxypropyl methylcellulose particles; and ii) at least one chemical agent selected from signaling agents; and consisting of only two or more components, the particles of the homogenized dry powder have an average particle size in the range of 80 μm to 101 μm; the signaling agent comprises garlic; The composition is for use as a nasally administered pharmaceutical.

2. 1. A composition in the form of a homogenized dry powder for use in the prevention and / or treatment of coronavirus diseases, said composition comprising: i) hydroxypropyl methylcellulose particles; and ii) at least one chemical agent selected from signaling agents; and iii) one or more biologically active agents; and consisting of only two or more components, the particles of the homogenized dry powder have an average particle size in the range of 80 μm to 101 μm; the signaling agent comprises garlic; the biologically active agent is an herbal or homeopathic agent selected from St. John's wort, valerian extract, ginkgo biloba extract, vitamin A, vitamin E, vitamin C, garlic, one or more probiotics, ginger, ellagic acid, echinacea, Swedish flower pollen, black walnut shells, lemongrass, mugwort, grapefruit seed extract, broccoli, digestive enzymes, hyaluronic acid, astragalus, rose hips, gentian, hypericum, horse chestnut, ginseng, green tea, phosphatidylserine, phosphatidylcholine, citrus fruits, pycnogenol, caffeine, quercetin, coenzyme Q10, yarrow, tea tree, noni juice, lipase, fructooligosaccharides, inulin, black cumin, stabilized allicin, and any combination thereof; The composition is for use as a nasally administered pharmaceutical.

3. 3. The composition of claim 1, wherein the signaling agent comprises 0.25% to 10% of the total weight of the composition.

4. The composition of claim 3, wherein the hydroxypropyl methylcellulose particles comprise 90% to 99.75% of the total weight of the composition.

5. The composition according to any one of claims 1 to 4, wherein the coronavirus is selected from SARS-CoV, MERS-CoV, SARS-COV-2, HCov-NL63, HCov-OC43, CoV-HKU1, HCov-229E and mutant strains thereof.

6. The composition of any one of claims 1 to 5, wherein the composition provides sustained release of the biologically active agent.

7. The composition of any one of claims 1 to 5, wherein the biologically active agent has a systemic effect when administered intranasally.

8. The composition of any one of claims 1 to 7, wherein the composition is for use in the treatment of COVID-19 disease.

9. 8. The composition of any one of claims 1 to 7, wherein the composition is for use in the prevention of COVID-19 disease.

10. 1) adding the powdered signaling agent to the hydroxypropyl methylcellulose particles; 2) diffusively blending the two components of 1) in a blender; 3) optionally adding powdered biologically active agent and further blending; 8. A method for manufacturing a composition according to any one of claims 1 to 7 for use as a medicament for treating COVID-19 disease, comprising:

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