Peptidase preparations for the treatment of microbial infections in the upper respiratory tract - Patent Application 20070233333

Abstract

The present disclosure relates to stable peptidase compositions and their use in the treatment and / or prevention of microbial infections.

Description

[Technical field]

[0001] The present invention relates to stable peptidase formulations for the treatment and prevention of microbial infections in the upper respiratory tract. [Background technology]

[0002] Upper respiratory tract infections are caused by acute infections involving the upper respiratory tract, including the nose, paranasal sinuses, pharynx, or larynx. Upper respiratory tract infections are one of the most common infections in the general population and cause colds and influenza, which are the main causes of unwanted work and school absenteeism. The majority of upper respiratory tract infections are caused by viruses. More than 200 different viruses have been isolated in patients with upper respiratory tract infections, including rhinoviruses, influenza viruses, respiratory syncytial viruses (RSV), coronaviruses, parainfluenza viruses, adenoviruses, enteroviruses, metapneumoviruses, and unknown viruses (Heikkinen et al, 2003).

[0003] In patients with SARS-CoV-2, nasal swabs yielded higher viral loads than throat swabs, suggesting that the nasal epithelium is the initial portal of infection and transmission (Sungnak et al., 2020). Given that nasal carriage is likely a key feature of SARS-CoV-2 transmission, drugs / vaccines administered intranasally could be highly effective in limiting the spread.

[0004] Viral infections can range from relatively mild, short-term infections, such as the common cold, or more severe viral infections, such as influenza, which spreads worldwide in seasonal epidemics, to aggressive, life-threatening infections. In addition to seasonal epidemics, several pandemics have occurred, the most devastating being the 1918 Spanish Flu, which killed 20–40 million people worldwide. Nearly half of the influenza-related deaths during the pandemic occurred among young, healthy adults, for reasons that remain unclear but are suspected to be related to secondary bacterial infections (Mallia and Johnston, 2007). Bacterial superinfections in viral respiratory diseases are well documented clinically, and physical damage to respiratory cells as a result of viral infection may lead to opportunistic bacterial attachment.

[0005] Given the magnitude of the health problems caused by colds, flu and influenza, along with newly emerging viral infections that are difficult to treat such as MERS (Middle East Respiratory Syndrome) and COVID-19, the need for novel therapies for viral infections is clear.

[0006] Vaccination is currently the primary focus for preventing the spread of influenza viruses, but due to mutations, new vaccines need to be developed every year. The development of antiviral drugs against the common cold is difficult due to the diversity of the viruses that cause this disease. Such therapeutics should have broad-spectrum activity in order to be suitable as antiviral countermeasures against a diverse range of viruses.

[0007] There are prophylactic drugs that have been proven to prevent cold infections, but these have side effects such as nasal bleeding. Other antiviral chemotherapeutic agents (e.g., ICAM-1 blockers, capsid binding agents, and protease inhibitors) have similarly failed to show a promising risk-benefit ratio (Allan et al., 2014).

[0008] There is an urgent need to develop new highly stable topical antibacterial compositions, such as antiviral compositions with broad spectrum activity (i.e., effective against a variety of enveloped and non-enveloped viruses, RNA and DNA-based genomes, and not dependent on different viral proteins (mutated or not)) that reduce and / or prevent viral binding to cells. Such compositions would ideally be based on non-toxic compounds. Moreover, such antiviral compositions should also be effective against secondary infections in the upper respiratory tract caused by bacteria and fungi, target microbial / bacterial adhesion without inducing resistance, and further have no or little effect on the natural microflora of the patient. Summary of the Invention

[0009] The inventors of the present invention have developed a novel formulation, which is a stable peptidase formulation for the prevention and treatment of viral infections in the upper respiratory tract. The formulation can be administered to the upper respiratory tract not only by nasal administration, thereby reaching the nasopharynx, but also by oral administration, so as to reach the pharynx.

[0010] This disclosure describes a product that acts as a complement to vaccines and systemic antivirals. It forms a thin, protective, broad-spectrum antiviral gel in the nasal passages and / or throat (pharynx) to prevent local uptake of viruses in the upper respiratory tract or reduce the spread of viral infection. Viruses are captured and bound to the gel, and then inactivated by broad-spectrum peptidases. The formulation constitutes an effective barrier and causes almost no stinging when applied to the nasal passages.

[0011] The formulation is capable of reducing SARS-CoV-2 virus by >99.9% in vitro.

[0012] The present invention is also active against bacteria and fungi by targeting the same virulence factor as viruses, namely the binding interaction between microorganisms and cells. The formulation does not induce microbial resistance and is therefore a good alternative to antibiotics. Thus, the formulations described herein have high potential for clinical use and provide a solution to the problem of eliminating the use of antibiotics.

[0013] In one aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: i. peptidases, ii. Stable compositions comprising sugar alcohols.

[0014] In another aspect, the present disclosure provides a method for producing a method for manufacturing a pharmaceutical composition comprising: i. peptidases, ii. glycerol, iii. xylitol, iv. hyaluronic acid, and v. A composition comprising a buffer.

[0015] In another aspect, the disclosure relates to such compositions for use in medicine.

[0016] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a pharmaceutical composition comprising: i. trypsin, ii. glycerol, iii. xylitol, iv. hyaluronic acid, v . CaCl 2 , and vi. Compositions comprising or consisting essentially of buffer solutions.

[0017] One aspect provides a composition disclosed herein for use in treating a condition selected from the group consisting of a microbial infection, a skin disease, and an oral disease in and / or on a mammal.

[0018] One aspect provides a composition as disclosed herein for use in the treatment and / or prevention of an upper respiratory tract infection, wherein the composition is formulated for nasal and / or oral administration.

[0019] In one aspect, the present disclosure provides a method for the prevention and / or reduction of a microbial infection, the method comprising administering a composition disclosed herein.

[0020] In yet another aspect, the present disclosure relates to a method for increasing microbial diversity, the method comprising administering a composition disclosed herein.

[0021] In one aspect, the present disclosure relates to a method of making the compositions disclosed herein, the method comprising: i. glycerol, ii. xylitol, iii.Buffer, iv. hyaluronic acid, and v. It involves mixing peptidases. [Brief description of the drawings]

[0022] [Figure 1] Enzyme activity as a function of dilution. The highest activity was observed at glycerol concentrations of 25-30 wt%. [Diagram 2] Effect of increasing carrageenan concentration on enzyme activity. Carrageenan concentrations >0.2mg / ml have an effect on activity. [Diagram 3] Enzyme stability in various formulations. Addition of >10% xylitol to 25-30% glycerol was required to achieve high stability. CX is a commercially available trypsin-containing mouth spray. [Figure 4] Enzyme stability in various formulations. Formulations N2 and N4 exhibited high stability without the addition of carrageenan. CX is a commercially available trypsin-containing mouth spray. [Diagram 5] Instantaneous viscosity of glycerol containing increasing concentrations of carrageenan. [Figure 6]Enzyme stability in xylitol-free TS01, TS02, TS02, and CX, which is a commercially available trypsin-containing mouth spray. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] definition As used herein, the term "microbial infection" refers to an infection by a microorganism, such as a fungus, bacteria, or virus. The term "viral infection" refers to any stage of a viral infection, including the latent stage, the latent or dormant stage, the acute stage, and the development and maintenance of immunity to the virus.

[0024] The "upper respiratory tract" includes the mouth, nose, sinuses, middle ear, throat, pharynx, larynx, and trachea.

[0025] As used herein, terms such as "preventing a viral infection" and "treating a viral infection" refer to inhibiting the replication of a particular virus, inhibiting viral transmission, or preventing a virus from establishing itself within its host, and ameliorating or alleviating symptoms of disease caused by a viral infection, such as sore throat, stuffy and / or runny nose, cough, and / or elevated temperature. Treatment is considered therapeutic if there is a reduction in viral load, and / or a reduction in morbidity and / or mortality.

[0026] The term "peptidase" as used herein includes proteases, proteinases and proteolytic enzymes, functional homologs or functionalized derivatives thereof, capable of cleaving short or long polypeptides of any length at peptide bonds, for example, by hydrolyzing the peptide bonds. Thus, the substrates of peptidases are polypeptides.

[0027] The term "sugar alcohol" as used herein refers to a sugar derivative, monosaccharide, disaccharide, or oligosaccharide in which the carbonyl group (aldehyde or ketone, reducing sugar) has been reduced to a primary or secondary hydroxyl group. Non-limiting examples of sugar alcohols include ethylene glycol (2 carbons), glycerol (3 carbons), erythritol (4 carbons), threitol (4 carbons), arabitol (5 carbons), xylitol (5 carbons), ribitol (5 carbons), mannitol (6 carbons), sorbitol (6 carbons), dolcitol (6 carbons), iditol (6 carbons), isomalt (12 carbons), maltitol (12 carbons), lactitol (12 carbons), polyglycitol, and other related polyol derivatives.

[0028] As used herein, the term "polyol" refers to a compound that contains multiple alcohol groups, such as propylene glycol. As used herein, a polyol does not necessarily have one hydroxy group on each carbon atom.

[0029] As used herein, the term "buffer" refers to an aqueous solution that contains an acid-base mixture for the purpose of stabilizing pH.

[0030] The term "hyaluronic acid" (HA) as used herein refers to a polymer comprising repeating disaccharide subunits of hyaluronan. The term also includes polymers in which the disaccharide subunits may be derivatized at one or more positions of the repeating D-glucuronic acid and / or D-N-acetylglucosamine subunits. Thus, the term hyaluronic acid encompasses hyaluronic acid, crosslinked forms of hyaluronic acid, derivatized forms, and pharmacologically acceptable salts thereof.

[0031] As used herein, the term "pharmacologically acceptable" generally refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with the tissues, organs, and / or body fluids of human beings and animals, within the scope of sound medical judgment, without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0032] The term "stability" as used herein can refer to in vivo stability or storage stability (eg, storage stability of a peptidase at room temperature).

[0033] As used herein, the term "flavoring agent" refers to any substance, natural or non-natural, capable of imparting a detectable flavor effect, particularly at concentrations of less than 4% by weight, more preferably less than 2% by weight.

[0034] The term "carrageenan" as used herein refers to a family of linear sulfated polysaccharides, usually extracted from red seaweed. All carrageenans are polysaccharides consisting of repeating galactose units, both sulfated and non-sulfated, and 3,6 anhydrogalactose. There are various types of carrageenans. The main types among them are kappa, lambda, and iota. Carrageenans are typically used for viscosity control or to achieve bioadhesion.

[0035] As used herein, the term "bioadhesion" refers to the ability of certain materials, such as polymers, to adhere to living or body tissue. As used herein, the term "mucoadhesion" refers to the ability of certain materials to adhere to mucosa or mucosal tissue (e.g., nasal mucosa) through the formation of chemical and / or physical bonds.

[0036] As used herein, the term "viscosity" refers to a fluid's resistance to flow. The unit of viscosity is Pascal seconds (Pa s or Pas).

[0037] Terms such as "oral," "oral," and "by oral administration," as used herein, refer to introducing a pharmaceutical composition into a subject via the oral cavity. In a preferred embodiment, the pharmaceutical composition is a liquid composition.

[0038] As used herein, terms such as "nasal" and "by nasal administration" refer to introducing a pharmaceutical composition into a subject via the nasal passages. In a preferred embodiment, the pharmaceutical composition is a liquid composition.

[0039] The term "tolerable" as used herein refers to, for example, irritation, stinging, and secretory irritation observed when the composition is used for nasal administration. Tolerability is herein rated from unacceptable to comfortable as assessed by the subject.

[0040] As generally used herein, the term "divalent cation" refers to a positively charged ion of any metal from the periodic table having a valence of 2. When the amount of a divalent cation, for example in an aqueous solution, is given as a percentage concentration, such as % w / w, the concentration of the divalent cation is based on the weight of the cation relative to the total weight of the composition of which the cation forms a part.

[0041] In the context of the present disclosure, the term "w / w" means "weight / weight". The expression "% w / w" is synonymous with "% by weight" or "percent by weight". By way of example, 10 g of a composition containing 50% w / w A contains 5 g of A.

[0042] In the context of the present disclosure, phrases such as "a composition comprising X to Y% A" are to be interpreted as meaning a composition comprising a range of X to Y% A, inclusive of both thresholds, i.e., the composition does not comprise less than X% A, and when X is less than Y, the composition does not comprise more than Y% A.

[0043] Nasal Anatomy and Tolerance The most common route of viral entry is through the respiratory tract. The respiratory tract refers to the pathway that carries air to the lungs and can be divided into upper and lower. The upper respiratory tract includes the nose, paranasal sinuses, throat (pharynx) and voice box (larynx). Mechanical barriers play an important role in the antiviral defense of the upper respiratory tract. For example, the airways are lined with mucociliary bodies, which consist of ciliated cells, mucus-secreting goblet cells, and subepithelial mucosecretory glands. Foreign particles deposited in the nasal cavity or upper respiratory tract become trapped in mucus and are carried to the back of the throat where they are swallowed. In the lower respiratory tract, particles trapped in mucus are lifted from the lungs to the throat by ciliary action. The lowest part of the airways, the alveoli, lack cilia or mucus, but macrophages lining the alveoli ingest and destroy particles. Other cellular and humoral immune responses are also mediated. To successfully infect the respiratory tract, a virus must not be cleared by mucus, neutralized by antibodies, or destroyed by alveolar macrophages.

[0044] A common factor shared by all cold viruses, and other infectious viruses of the respiratory tract, is that they must evade the mechanical defense mechanisms of the airways. Microbial attachment, in which microbial surface proteins recognize cell surface proteins, is the most common mode of establishing infection in the host (Meena et al., 2020).

[0045] The present invention aims to reinforce the natural mechanical barrier against microbial infection. Blocking or impeding the movement of viruses to reach specific host cell receptors is an attractive way to provide safe and effective protection against viruses.

[0046] For the best broad spectrum protection against viral infections in the upper respiratory tract, both the nasal and oral cavities should be protected.

[0047] In general, the mucous membrane of the nasal cavity is more sensitive to hyperosmolar solutions than the oropharynx. Local irritation and nasal burning / pain have been reported, for example, after intranasal administration of Nasalide® nasal spray, with up to 45% of patients experiencing nasal burning (Trangsrud et al, 2002).

[0048] The present invention allows for both rapid clearance and nasal sensitivity.

[0049] Peptidases The development of antiviral drugs against cold viruses is difficult due to the diversity of viruses that cause this disease. In the present disclosure, a peptidase composition for the prevention of microbial infection is used. As used herein, the term "peptidase" includes proteases, proteinases and proteolytic enzymes that can catalyze protein degradation in vivo in a mammalian (e.g., human) body, thereby cleaving short or long polypeptides of any length at peptide bonds, for example, by hydrolyzing the peptide bonds, as well as functional homologs or functionalized derivatives thereof. The substrate can be a naturally occurring or synthetic polypeptide. Peptidases can be either specific and capable of hydrolyzing only selected peptide bonds, or non-specific and capable of hydrolyzing many different peptide bonds.

[0050] Viruses and bacteria can be present in the nasopharynx without causing respiratory symptoms. The upper respiratory tract harbors a wide range of commensal and potentially pathogenic bacteria that form a complex microbial community. This community appears to be constantly subject to synergistic and competitive interspecies interactions. Disturbance of the equilibrium, for example by the acquisition of new bacteria or viruses, can lead to overgrowth and invasion.

[0051] Peptidases are highly suitable for reducing or inhibiting unwanted microbial adhesion, such as viral adhesion, and furthermore, peptidases are non-toxic, well tolerated and do not induce resistance. Compositions containing peptidases can be used as therapeutic agents as an alternative to antibiotics in the treatment of microbial infections without interfering with the desired healthy microflora.

[0052] Stable peptidase compositions The present disclosure relates to a stable peptidase composition for the prevention and / or treatment of microbial infections. The composition provides a means of strengthening the intrinsic barrier to enhance protection against viruses. The disclosed composition comprises a peptidase capable of hydrolyzing microbial surface proteins, thereby preventing microbial attachment and the establishment of a microbial infection. Additionally, the composition comprises a mixture of sugar alcohols or polyols that can provide a stable composition that is tolerable for intranasal administration.

[0053] The compositions of the present disclosure are more effective against viruses and have improved shelf life and activity compared to other known peptidase preparations (e.g., commercially available peptidase preparations).Furthermore, the compositions are suitable for nasal administration to humans and other mammals (as concluded from disclosed Example 4), which is a significant improvement over other known peptidase preparations that are only tolerated by oral administration.

[0054] Experimental results show that the compositions of the present disclosure are highly effective in reducing microbial infections. In a virucidal efficacy test (results presented in Example 5), both selected formulations (for throat and nose) were found to be more effective against the SARS-CoV-2 virus in in vitro assays compared to a commercial mouth spray containing trypsin.

[0055] One embodiment of the present disclosure comprises: i. 0.005-1.0% w / w of a peptidase; ii. Providing a composition comprising 1.0-70% w / w sugar alcohol.

[0056] In one embodiment, the composition comprises: i. 0.005-1.0% w / w of a peptidase; ii. 20-70% w / w glycerol, and iii. Contains 1.0-65% w / w xylitol.

[0057] In one embodiment, the composition comprises: i. 0.005-1.0% w / w of a peptidase; ii. 20-70% w / w glycerol, and iii. Contains 1.0-65% w / w xylitol; The combination of components in the composition does not exceed 100%.

[0058] In one embodiment, the composition comprises 0.005-1.0% w / w peptidase, such as 0.01-0.075% w / w peptidase, for example, 0.015-0.055% w / w peptidase.

[0059] In one embodiment, the peptidase is selected from the group consisting of serine proteases (such as trypsin and chymotrypsin), threonine proteases, cysteine ​​proteases, aspartic acid proteases, glutamic acid proteases and metalloproteases, functional homologues thereof, and functionalized derivatives thereof, hi one embodiment, the peptidase is a single type of peptidase or a mixture of different types of peptidases.

[0060] In one embodiment of the present disclosure, the combination of components of the composition does not exceed 100%.

[0061] Peptidase activity Peptidase activity can be determined by measuring the product formed under given conditions per amount of peptidase for a set time. The specific activity of a peptidase is the enzyme activity per protein concentration. Thus, the total peptidase activity is determined by the specific activity and purity. For example, the desired activity can be achieved by using a relatively impure peptidase at a relatively high concentration, or by using a relatively pure peptidase at a relatively low concentration. The optimal concentration of the peptidase used can be evaluated by measuring the enzyme activity.

[0062] Trypsin The peptidase may be a protease such as trypsin, which cleaves peptide chains primarily at the carboxyl side of the amino acids lysine and arginine.

[0063] In one embodiment, the peptidase is a protease. In another embodiment, the protease is trypsin. In yet another embodiment, the protease is a functional homolog or functionalized derivative of trypsin that has proteolytic activity.

[0064] In one embodiment of the disclosure, the composition comprises 0.005-1.0% w / w of the protease, e.g., 0.005-1.0% w / w of the protease. In a further embodiment of the disclosure, the composition comprises 0.015-0.055% w / w of the protease. In a particular embodiment of the disclosure, the composition essentially comprises 0.015-0.055% w / w of the protease, e.g., 0.015-0.055% w / w of the protease.

[0065] In one embodiment of the present disclosure, the composition comprises 0.005-1.0% w / w trypsin, a functional homologue or a functionalized derivative thereof, for example, 0.005-1.0% w / w trypsin, a functional homologue or a functionalized derivative thereof.

[0066] In further embodiments of the disclosure, the composition comprises 0.015-0.055% w / w trypsin, a functional homologue or a functionalized derivative thereof. In certain embodiments of the disclosure, the composition essentially comprises 0.015-0.055% w / w, e.g., 0.015-0.055% w / w trypsin, a functional homologue or a functionalized derivative thereof.

[0067] Cations such as calcium The compositions of the present disclosure are stable peptidase formulations. The peptidase may be stabilized by a salt.

[0068] In one embodiment, the composition further comprises a salt, hi another embodiment, the composition comprises a divalent cation.

[0069] As used herein, the term "divalent cation" refers to a positively charged ion of any metal from the periodic table that has a valence of two, such as calcium.

[0070] In one embodiment, the present invention discloses a composition comprising trypsin, the composition further comprising a salt, hi another embodiment, the present invention discloses a composition comprising trypsin, the composition further comprising a salt with a divalent cation.

[0071] In a more specific embodiment, the present invention discloses a composition comprising a peptidase, the composition further comprising a salt having a divalent cation, for example, 0.003-0.2% w / w of a salt comprising a divalent cation, for example, 0.003-0.15% w / w, for example, 0.003-0.1% w / w of a salt comprising a divalent cation.

[0072] In another more specific embodiment, the composition comprising trypsin, a functional homologue thereof, or a functionalized derivative thereof further comprises 0.003 to 0.2% w / w of a salt comprising a divalent cation, such as 0.003 to 0.15% w / w, for example 0.003 to 0.1% w / w of a salt comprising a divalent cation.

[0073] In one embodiment of the present disclosure, the divalent cation is calcium (Ca 2+ In another embodiment, the salt containing a divalent cation is a pharma- ceutically acceptable salt of calcium, including hydrates.

[0074] In another embodiment, the pharma- ceutically acceptable salt of calcium is CaCl 2 In yet another embodiment, the salt containing a divalent cation is a pharma- ceutically acceptable inorganic salt of calcium, such as calcium chloride dihydrate, including hydrates thereof. In another embodiment, the salt containing a divalent cation is a pharma- ceutically acceptable organic salt of calcium, such as calcium acetate or calcium citrate.

[0075] In one embodiment, the composition comprises 0.003-0.2% w / w calcium, such as 0.003-0.15% w / w, for example 0.003-0.1% calcium.

[0076] In one embodiment, the composition further comprises 0.003-0.2% w / w calcium ions, for example, 0.003-0.15% w / w calcium ions, for example, 0.003-0.1% w / w calcium ions. Calcium ions are Ca 2+ means...

[0077] In another embodiment, the composition comprises 0.002-0.2% w / w calcium chloride, such as 0.005-0.02% w / w calcium chloride, for example 0.010-0.015% w / w calcium chloride.

[0078] In yet another embodiment, the composition comprises 0.002-0.2% w / w calcium chloride dihydrate, such as 0.005-0.02% w / w calcium chloride dihydrate, for example, 0.010-0.015% w / w calcium chloride dihydrate.

[0079] In one embodiment of the present disclosure, the composition comprises: i. 0.005-1.0% w / w trypsin; ii. 1. 0-70% w / w of a sugar alcohol, and iii. Contains 0.010-0.015% w / w of calcium dichloride dihydrate.

[0080] Sugar alcohols / polyols The composition of the present disclosure is a stable peptidase formulation. Inactivation of peptidase improves its stability in the composition. One way to inactivate peptidase is by reducing the water activity of the composition. The water activity of an aqueous solution can be reduced by the presence of ions and / or other molecules.

[0081] The present invention includes salts, sugar alcohols, and / or polyols, which act in part to reduce the water activity of the composition. Reducing the water activity inactivates peptidases. Furthermore, it is beneficial for the purposes of the present disclosure that the composition is hyperosmolar. Hyperosmolar compositions generally stimulate nasal secretion, which has a flushing effect on the nasal mucosa, cleaning the infected surface of all contaminants, such as viral particles and free-floating bacteria, without any toxic effect on cells or cell matrix.

[0082] In one embodiment of the disclosure, the composition includes one or more of a diol, triol, or polyol, such as glycerol, propylene glycol, or a sugar alcohol.

[0083] In another embodiment, the composition comprises 35-70% w / w sugar alcohol, such as 40-65% w / w sugar alcohol, for example 45-60% w / w sugar alcohol.

[0084] The sugar alcohol can be glycerol, erythritol, xylitol, mannitol, arabitol, maltitol, lactitol, isomalt, sorbitol, hydrogenated starch hydrolysates or propylene glycol, and any other suitable polyol or mixtures thereof that provide a stable solution of the peptidase.

[0085] Although the oral cavity tolerates high concentrations of glycerol well, high concentrations of glycerol have a harsh stinging effect on the thinner and more sensitive nasal mucosa. A composition containing 50% glycerol cannot be sprayed into the nasal cavity due to the severe stinging caused by the hyperosmotic effect of glycerol. 10-25% glycerol can be tolerated in small amounts. However, at low glycerol concentrations (25%) after dilution, the enzymes disclosed herein are not stable for long-term storage, significantly reducing their use in the treatment and / or prevention of infection.

[0086] Because the nasal mucosa is more sensitive to hypertonic solutions compared to the oropharynx, the glycerol concentration in the nasal formulation must be diluted compared to oral formulations while still maintaining a stable composition.

[0087] Various combinations and concentrations of sugars and sugar alcohols were screened for applicability to minimize nasal irritation and it was surprisingly found that the disclosed compositions comprising a mixture of the sugar alcohols glycerol and xylitol provide stable compositions that are tolerable for nasal administration.

[0088] Xylitol is a sugar alcohol with properties similar to those of glycerol. Xylitol can have a protein stabilizing effect. Xylitol also has a sweetness intensity equal to that of sucrose.

[0089] In one embodiment, the composition comprises 20-70% by weight glycerol, for example 25-65% by weight glycerol, in one particular embodiment, the composition comprises 27-57% by weight glycerol.

[0090] In another embodiment, the composition comprises 1.0 to 65% w / w xylitol, such as 3.0 to 50% w / w xylitol, for example 4.5 to 30% w / w xylitol.

[0091] In yet another embodiment the composition comprises a combined amount of glycerol and xylitol of at least 40% w / w, such as at least 45% w / w, such as at least 50% w / w.

[0092] In a preferred embodiment, the composition of the present invention comprises i. 0.005-1.0% w / w of a peptidase; ii. 20-70% w / w glycerol, and iii. Contains 1.0-65% w / w xylitol.

[0093] The taste and tolerability of the product are particularly important for patient compliance when taking a nasal spray. The present invention presents a tolerable formulation with a sweet and pleasant taste.

[0094] buffer solution The term "buffer" is intended to mean an aqueous solution containing an acid-base mixture for the purpose of stabilizing the pH, which is important for both the stability of the peptidase, the activity of the peptidase, and for the tolerability of the composition when used to treat microbial infections in the oral and nasal cavities.

[0095] Examples of buffers are Trizma, Bicine, Tricine, MOPS, MOPSO, MOBS, Tris, Hepes, HEPBS, MES, phosphate, carbonic acid, acetate, citric acid, glycolic acid, lactic acid, boric acid, ACES, ADA, tartaric acid, AMP, AMPD, AMPSO, BES, CABS, cacodylic acid, CHES, DIPSO, EPPS, ethanolamine, glycine, HEPPSO, imidazole, imidazole lactate, PIPES, SSC, SSPE, POPSO, TAPS, TABS, TAPSO, and TES. The buffer can be a mixture of two or more buffer systems. In one embodiment, the buffer is any of the buffers listed above, including a mixture of two or more buffers. In one embodiment of the present disclosure, the composition includes a buffer that stabilizes the pH of the composition. The buffer may be selected from the group consisting of tris(hydroxymethyl)aminomethane (Tris), 3-(N-morpholino)propanesulfonic acid (MOPS), phosphate, or any other suitable buffer, such as a pharma- ceutically acceptable buffer.

[0096] In one embodiment, the composition of the present disclosure comprises 0.01 to 2.4% w / w buffer, such as 0.07 to 0.3% w / w buffer, for example, 0.09 to 0.2% w / w buffer.

[0097] In another embodiment, the composition comprises a buffer concentration in the range of 1-200 mM, such as 5-25 mM, for example, 8-15 mM.

[0098] The pH of the composition is adjusted so as to be suitable for use in each of the nasal cavity and the oral cavity. The nasal cavity has a pH range of 5.5 to 6.5. The throat has a pH range of 7.2 to 8.5. The composition has a pH range of 5.5 to 8.5, for example, 6.0 to 7.0, for example, 6.3 to 6.7.

[0099] In one embodiment, the composition has a pH in the range of 6.2 to 7.0, for example, 6.4 to 6.6, In another embodiment, the composition has a pH in the range of 7.2 to 8.5, for example, 7.4 to 7.6.

[0100] The pH of a solution can be assessed in a number of ways. For example, the pH can be assessed using a pH electrode or a pH indicator.

[0101] In a preferred embodiment, the composition comprises: i. 0.005-1.0% w / w of a peptidase; ii. 20-70% w / w glycerol; iii. 1. 0-65% w / w xylitol, and iv. Contains 0.01-1.2% w / w buffer.

[0102] In one embodiment the composition further comprises at most 50% w / w water, such as at most 40% w / w, such as at most 30% w / w water.

[0103] polymer For mucosal application, the composition may advantageously contain ingredients that impart mucoadhesive properties to the composition.

[0104] The disclosed compositions may include a polymer to increase the residence time of the composition (e.g., gel). In certain embodiments, the polymer(s), relative amount, and concentration are selected to provide a film that is mucoadhesive. Mucoadhesive performance is typically observed in polymers that have charged groups or non-ionic functional groups that can form hydrogen bonds. Negatively charged polysaccharides, such as hyaluronate salts of hyaluronic acid, and sulfated polysaccharides, such as carrageenan, have been found to be effective.

[0105] In one embodiment of the present disclosure, the composition comprises a polymer that can increase the residence time of the composition in the mucosa. In one embodiment, the composition comprises a polymer selected from the group consisting of hyaluronic acid and hyaluronic acid derivatives. Such polymers function in the nasal cavity as a means of strengthening the gel, increasing mucoadhesion, and reducing irritation.

[0106] In one embodiment, a polymer is included to strengthen the composition by forming a gel and reduce the potential for irritation in the nasal passages.

[0107] Hyaluronic acid In one embodiment of the present disclosure, the composition comprises hyaluronic acid.

[0108] In one embodiment, the composition comprises hyaluronic acid, for example, 0.0001-2.0% w / w hyaluronic acid, for example, 0.001-1.5% w / w, such as 0.01-1.0% w / w, for example, 0.02-0.5% w / w hyaluronic acid.

[0109] In one embodiment of the present disclosure, the composition comprises: i. 0.005-1.0% w / w of a peptidase; ii. 20-70% w / w glycerol; iii. 1.0-65% w / w xylitol; iv. 0.0001-1.5% w / w hyaluronic acid, and v. 0.01-1.2% w / w buffer solution or consisting essentially of same.

[0110] In another embodiment of the present disclosure, the composition comprises: i. 0.005-1.0% w / w trypsin; ii. 20-70% w / w glycerol; iii. 1.0-65% w / w xylitol; iv. 0.0001-1.5% w / w hyaluronic acid; v. 0.002 to 0.2% w / w CaCl 2 , and vi. Containing or consisting essentially of 0.01-1.2% w / w buffer solution.

[0111] In another embodiment, the composition comprises: i. 0.015-0.055% w / w trypsin; ii. 27-57% w / w glycerol; iii. 4.5-30% w / w xylitol; iv. 0.0002-0.02% w / w hyaluronic acid; v. 0.010-0.015% w / w CaCl 2 , and vi. Containing or consisting essentially of 0.1-0.14% w / w Tris buffer.

[0112] In one embodiment, the pH of the composition is 5.5 to 8.5, for example, 6.0 to 7.0, for example, 6.3 to 6.7. In one embodiment, the pH of the composition is 6.2 to 7.0, for example, 6.4 to 6.6. In one embodiment, the pH of the composition is 7.2 to 8.5, for example, 7.4 to 7.6.

[0113] In one embodiment, the composition comprises water. In certain embodiments of the present disclosure, at least a portion of the water in the composition is derived from a buffer. The presence of free water may activate and / or destabilize peptidases. Water in the composition may be inactivated by other ingredients, such as, for example, glycerol and other alcohols as described herein. Thus, in one embodiment, the composition comprises at most 50% w / w water, such as at most 40% w / w water, such as at most 30% w / w water. To improve the stability of peptidases, it is important that the water activity in the composition is low. When the composition is applied, for example, to the nasal cavity, oral cavity, wound, or skin, water from the nasal cavity, oral cavity, wound, or skin mixes with the composition, increasing the water activity. This activates the peptidases.

[0114] In another embodiment, the composition is in the form of a spray, lozenge, troche, chewing gum, gel, or liquid.

[0115] Carrageenan In one embodiment, the composition comprises carrageenan.

[0116] Carrageenans are typically used for viscosity control or to achieve bioadhesion. The term "carrageenan" refers to a family of linear sulfated polysaccharides that are usually extracted from red seaweed. Carrageenans are polysaccharides composed of both sulfated and non-sulfated repeating galactose units and 3,6 anhydrogalactose.

[0117] In one embodiment the composition comprises no more than 1% w / w carrageenan, such as no more than 0.5% w / w carrageenan, such as no more than 0.1% w / w carrageenan.

[0118] Flavoring Agent For the compliance of patients who receive the disclosed compositions, the aroma and taste of the product are particularly important.The present invention presents a tolerable formulation with a sweet and pleasant taste, due in part to the presence of xylitol.If the composition further comprises a flavoring agent, the aroma and taste can be further improved.

[0119] The term "flavoring agent" refers to any substance, natural or non-natural, capable of exerting a detectable flavor effect, particularly at concentrations of less than 4% w / w, more preferably less than 2% w / w. Suitable flavors or flavoring agents include, but are not limited to, eucalyptus, mints such as peppermint and spearmint, menthol, chocolate, licorice, citrus and other fruit flavors, gamma octalactone, vanillin, ethyl vanillin, breath freshener flavors, spicy flavors such as cinnamon, methyl salicylate, linalool, bergamot oil, geranium oil, lemon oil, and ginger oil.

[0120] In one embodiment, the composition comprises a flavoring agent.

[0121] In another embodiment, the flavoring agents are natural or non-natural, eg, synthetic.

[0122] In yet another embodiment, the natural or non-natural flavoring agent is spearmint or eucalyptus.

[0123] In a further embodiment the composition comprises 0.01 to 0.4% w / w flavouring, such as 0.02 to 0.1% w / w flavouring, for example 0.035 to 0.055% w / w flavouring compound.

[0124] In one embodiment, the composition comprises: i. 0.02-0.05% w / w of a peptidase; ii. 29-56% w / w glycerol; iii. 4 to 25% w / w xylitol; iv. 0.0002-0.0004% w / w hyaluronic acid; v. 0.1-0.2% w / w buffer solution, vi. 0 to 0.1% w / w of carrageenan, and vii. Comprising or consisting essentially of 0-0.05% w / w flavoring agent.

[0125] formulation Disclosed herein is a stable composition for the treatment of microbial infections, particularly viral infections in the upper respiratory tract.

[0126] Viral replication occurs in the upper respiratory tract. The currently developed oral sprays reach the oropharynx but do not reach the nasal cavity, including the nasopharynx, via the oral route of administration. It is therefore highly desirable to develop a sprayable composition that can be administered nasally, so that the nasal cavity and nasopharynx are reached using a nasal spray. In order for the composition to be sprayable, it is preferred that the composition has a viscosity that is at most 0.015 Pa·s. Administration to the upper respiratory tract is necessary for the composition to be active against microbial infections in the upper respiratory tract.

[0127] In one embodiment, the composition is formulated as a spray, lozenge, troche, chewing gum, gel, or liquid.

[0128] In one embodiment, the composition is a sprayable composition. In another embodiment, the composition is in the form of a spray. In a preferred embodiment, the composition is a nasal spray or a throat spray.

[0129] In order to administer a composition using a nasal or throat spray, the viscosity of the composition must allow the composition to be sprayable.

[0130] In one embodiment, the composition has a viscosity of at most 0.015 Pa.s. This is supported by the findings of Example 6 disclosed herein.

[0131] In comparison to oral sprays, nasal sprays target the nasal cavity, which is the primary site of upper respiratory tract infection.

[0132] medical use Disclosed herein is a stable composition for the treatment of microbial infections, particularly viral infections in the upper respiratory tract.

[0133] The composition of the present disclosure can complement vaccines and systemic antiviral drugs. It forms a thin, protective, broad-spectrum antiviral gel in the nasal passages and throat, preventing or reducing the spread of viral infection. The virus is captured, bound to the gel, and then inactivated by broad-spectrum peptidases. Compared to antiviral drugs that act systemically and intracellularly to inhibit the replication system, the composition of the present disclosure reduces and / or prevents local uptake of the virus in the upper respiratory tract. These mechanisms of action complement each other to achieve more efficient treatment and / or prevention of infection. The present invention is also active against bacteria and fungi by targeting the same virulence factors as viruses, namely the proteins responsible for the attachment between microorganisms and cells.

[0134] In one embodiment, the viral infection is selected from the group consisting of cold, pneumonia, bronchitis, Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), sinusitis, otitis media, and pharyngitis. One embodiment provides a method for treating and / or preventing a viral infection in a mammal comprising administering to a subject a therapeutically effective amount of a polypeptide having peptidase activity.

[0135] The common cold is a mild acute respiratory infection characterized by sore throat, malaise, rhinorrhea, nasal congestion, headache, coughing, sneezing, and sometimes mild fever (Jackson et al, 1958). Influenza is a more severe viral infection that spreads worldwide in seasonal epidemics, infecting up to 20% of the population and can cause substantial mortality depending on the circulating virus.

[0136] In one embodiment, the compositions of the present disclosure are for medical use.

[0137] In one embodiment, the present disclosure relates to a composition for use in the treatment and / or prevention of a microbial infection.

[0138] In one embodiment, the present disclosure relates to a composition for use in the treatment and / or prevention of a viral infection, a bacterial infection, a fungal infection, and / or a yeast infection.

[0139] In one embodiment, the present disclosure relates to a composition for use in the treatment and / or prevention of a viral infection.

[0140] In one embodiment, the present disclosure relates to a composition for use in the treatment and / or prevention of a viral infection from a virus selected from the group consisting of rhinovirus, influenza virus such as influenza A virus, respiratory syncytial virus (RSV), coronavirus, parainfluenza virus, adenovirus, enterovirus, metapneumovirus, and other infectious viruses.

[0141] In one embodiment, the present disclosure relates to a composition for use in the treatment and / or prevention of an infectious disease, such as cold, influenza, rhinitis, sinusitis, bronchitis, Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), coronavirus disease 2019 (COVID-19), pneumonia, viral meningitis, herpangina, herpes virus, papilloma virus, or any other disease caused by a viral infection.

[0142] In one embodiment, the present disclosure relates to a composition for use in the treatment and / or prevention of a viral upper respiratory tract infection.

[0143] In one embodiment, the present disclosure relates to a composition for use in the treatment and / or prevention of cold, influenza, rhinitis, sinusitis, a viral upper respiratory tract infection resulting in COVID-19, or other disease caused by a viral upper respiratory tract infection.

[0144] In one embodiment, the compositions disclosed herein are for use in the prevention and / or treatment of symptoms associated with a viral infection, such as sore throat, fatigue, runny nose, stuffy nose, headache, cough, sneezing, and / or fever.

[0145] In one embodiment, the compositions disclosed herein are for use in the prevention and / or treatment of a disease, disorder or condition selected from the group consisting of microbial infection and / or oral disease.

[0146] In one embodiment, the compositions disclosed herein are for use in the prevention and / or treatment of a microbial infection selected from the group consisting of a viral infection, a bacterial infection, a fungal infection, and a yeast infection.

[0147] In another embodiment, the compositions disclosed herein are for use in the prevention and / or treatment of microbial infections and / or oral diseases, such as periodontal disease, in and / or for a mammal.

[0148] In another embodiment, the compositions disclosed herein are for use in the prevention and / or treatment of microbial infections and / or oral diseases, such as gingivitis, in and / or to a mammal.

[0149] In one embodiment, the present disclosure provides a method for the preparation of a pharmaceutical composition comprising administering to a patient a therapeutically effective amount of i. peptidases, ii. glycerol, iii. xylitol, iv. hyaluronic acid, and v. Providing a composition comprising a buffer. In one embodiment, the peptidase of the composition is trypsin. In one embodiment, the composition comprising trypsin is calcium (Ca 2+ ).

[0150] One embodiment provides a method for the prevention and / or treatment of a disease, disorder or condition selected from the group consisting of microbial infections and / or oral diseases, comprising administering to said patient a composition comprising: i. peptidases, ii. glycerol, iii. xylitol, iv. hyaluronic acid, and v. Providing a composition comprising a buffer.

[0151] In one embodiment, the microbial infection and / or oral disease is gingivitis or periodontal disease in and / or for a mammal.

[0152] One embodiment is a method for the prevention and / or treatment of a disease, disorder or condition selected from the group consisting of microbial infections and oral diseases, comprising administering to a subject a composition comprising the composition a composition comprising: i. 0.005-1.0% w / w of a peptidase; ii. 20-70% w / w glycerol; iii. 1.0-65% w / w xylitol; iv. 0.0001-1.5% w / w hyaluronic acid, and v. A composition comprising 0.01 to 1.2% w / w of a buffer solution is provided.

[0153] One embodiment is a method for the prevention and / or treatment of a disease, disorder or condition selected from the group consisting of microbial infections and oral diseases, comprising administering to a subject a composition comprising the composition a composition comprising: i. 0.015-0.055% w / w trypsin; ii. 27-57% w / w glycerol; iii. 4.5-30% w / w xylitol; iv. 0.0002-0.02% w / w hyaluronic acid; v. 0.010-0.015% w / w CaCl 2 , and vi. Provide a composition comprising 0.1 to 0.14% w / w of a buffer solution.

[0154] One embodiment provides a composition as disclosed herein for use in the prevention and / or treatment of a microbial infection selected from the group consisting of a viral infection, a bacterial infection, a fungal infection, and a yeast infection.

[0155] One embodiment provides a composition disclosed herein for use in the prevention and / or treatment of a microbial infection that is a viral infection. In another embodiment, the composition is for use in the treatment of a viral upper respiratory tract infection. In another more specific embodiment, the microbial infection is a viral upper respiratory tract infection.

[0156] One embodiment provides a composition disclosed herein for use in the prevention and / or treatment of cold, influenza, rhinitis, sinusitis, an upper respiratory tract infection resulting in COVID-19, or another disease caused by an upper respiratory tract infection.

[0157] One embodiment provides a composition disclosed herein for use in the prevention and / or treatment of a viral infection from a virus selected from the group consisting of rhinovirus, influenza virus, respiratory syncytial virus (RSV), coronavirus, parainfluenza virus, adenovirus, enterovirus, metapneumovirus, and other infectious viruses.

[0158] In one embodiment, other infectious viruses that can infect and / or enter cells include Corynebacterium diphtheriae, Lassa fever virus (arenavirus), Astrovirus, Hantavirus, Rift Valley fever virus (phlebovirus), Calicivirus, Ebola virus, Marburg virus, Japanese encephalitis virus, Dengue virus, Yellow fever virus, Hepatitis C virus, Hepatitis G virus, Hepatitis B virus, Hepatitis D virus, Herpes simplex virus, Cytomegalovirus, and Hepatitis B virus. Viruses that cause infection include rabies, Epstein-Barr virus, varicella-zoster virus, human herpes virus, rubella virus, mumps virus, morbillivirus, measles virus, papillomavirus, JC virus (polyomavirus), BK virus (polyomavirus), parvovirus, coxsackievirus (A and B), hepatitis A virus, poliovirus, reovirus, rabies virus (lyssavirus), human immunodeficiency virus 1 and 2, and human T-cell leukemia virus.

[0159] One embodiment provides a composition disclosed herein for use in the prophylaxis and / or treatment of a viral infection causing an infectious disease such as cold, influenza, rhinitis, sinusitis, bronchitis, Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), coronavirus disease 2019 (COVID-19), pneumonia, viral meningitis, herpangina, herpes virus, papilloma virus, or any other disease caused by a viral infection.

[0160] One embodiment provides a composition disclosed herein for use in the prevention and / or treatment of symptoms associated with a viral infection, such as sore throat, fatigue, runny nose, stuffy nose, headache, cough, sneezing, and / or fever.

[0161] One embodiment provides a composition disclosed herein for use in the manufacture of a medicament for the prevention and / or treatment of a disease, disorder, or condition selected from the group consisting of a microbial infection, a skin disease, and an oral disease.

[0162] In one embodiment, the composition is for oral use, hi another embodiment, the composition is for nasal use.

[0163] In one embodiment, the nasal composition has a pH in the range of 6.2 to 7.0, for example, 6.4 to 6.6, in another embodiment, the oral composition has a pH in the range of 7.2 to 8.5, for example, 7.4 to 7.6.

[0164] One embodiment provides a method for treating or preventing a disease, disorder, or condition selected from the group consisting of a microbial infection, a skin disease, and an oral disease, the method comprising administering a composition disclosed herein to an individual in need thereof.

[0165] One embodiment provides a method for treating or preventing a disease, disorder or condition selected from the group consisting of a microbial infection, a skin disease, and an oral disease, the method comprising: i. peptidases, ii. glycerol, iii. xylitol, iv. hyaluronic acid, and v. Administering a composition comprising a buffer.

[0166] In one embodiment, the peptidase of the composition is trypsin. In one embodiment, the composition comprising trypsin is calcium (Ca 2+ ).

[0167] One embodiment provides a method of preventing and / or reducing a viral infection, the method comprising administering to a subject in need thereof a composition disclosed herein.

[0168] One embodiment provides a method of increasing microbial diversity, the method comprising administering a composition, comprising administering a composition disclosed herein to a subject in need thereof.

[0169] One embodiment provides a method for producing a composition disclosed herein, the method comprising: i. Sugar alcohols ii.Buffer, iii. hyaluronic acid, and iv. Including mixing with peptidase.

[0170] A further embodiment is i. carrageenan, and / or ii. A method for making the compositions disclosed herein is provided, further comprising mixing a flavoring agent.

[0171] One embodiment provides a method of increasing microbial diversity, the method comprising administering a composition disclosed herein. EXAMPLES

[0172] Example 1 Enzyme activity as a function of dilution in Tris-HCl buffer The purpose of the experiment was to measure the enzyme activity as a function of dilution in Tris-HCl buffer. In addition, the effect of carrageenan concentration on the enzyme activity was evaluated.

[0173] method Seven different formulations with glycerol concentrations ranging from 40 to 15% by weight were prepared along with formulations with increasing carrageenan concentrations. Enzyme activity was measured. Samples were removed at given time points and analyzed directly by measuring the enzyme activity. Activity was measured at 30 °C using Z-Gly-Pro-Arg-pNA as substrate in a spectrophotometer at 405 nm. The decrease in absorbance per minute was used as the activity parameter. The remaining activity relative to the zero time point was determined and used to describe stability.

[0174] Results and Conclusions The results of glycerol dilution are shown in Figure 1. Enzyme activity increases upon dilution with buffer. Absolute maximum activity was obtained with 25-30% glycerol.

[0175] The results for formulations with increasing concentrations of added carrageenan (0-0.8 mg / ml) are shown in Figure 2. Concentrations >0.2 mg / ml appear to reduce activity.

[0176] Example 2 Stability of Nasal Spray Formulations The purpose of the experiment was to determine the enzyme stability in various nasal spray formulations.

[0177] method The various combinations were mixed and the pH was adjusted to pH 6.5 using Tris buffer. The formulations described in Table 1 were tested. [Table 1]

[0178] The formulations were placed in sealed polypropylene bottles. The stability of the products from each formulation composition was tested by placing them under stress conditions (40±2°C / 100% RH) for up to 115 days. Enzyme activity was measured using the same method as described in Example 1.

[0179] Results and Conclusions The results are shown in Figure 3. The addition of >10% xylitol to 25-30% glycerol resulted in increased stability, as exemplified by formulations NO3, NO4 and NO5.

[0180] These findings suggest the possibility of replacing part of the glycerol with xylitol to improve the stability of the enzyme under stress conditions at 40 °C. The expected shelf life of the formulation containing 30% glycerol (formulation NO1) is <3-6 months at 25 °C, while the expected shelf life of NO5 is expected to be >2 years. The presence of excessive amounts of carrageenan reduced the stability.

[0181] Example 3. Stability of Throat Spray Formulation The purpose of the experiment was to determine the enzyme stability in various throat spray formulations.

[0182] method Various formulations were tested, as described in Table 2. The pH was adjusted to pH 7.5 using Tris buffer. [Table 2]

[0183] The formulations were stored in sealed polypropylene bottles. The stability of the products from each formulation composition was tested by placing them under stress conditions (40±2°C / 100% RH) for up to 115 days. Enzyme activity was measured using the same method as described in Example 1.

[0184] Results and Conclusions The results are shown in Figure 4. Formulations N2 and N4 showed the highest stability. The addition of 0.5% carrageenan decreased the stability, but the presence of hyaluronic acid and xylitol showed no adverse effect on the stability.

[0185] Example 4. Tolerability of the nasal spray formulation The purpose of the study was to evaluate the tolerability, irritation / stinging, and experienced secretory irritation of various nasal spray formulations by spraying the formulations into the nasal cavity. The nasal sprays are hyperosmolar and are therefore intended to stimulate nasal secretions with a flushing effect on the nasal mucosa. To minimize nasal irritation, the glycerol concentration in the nasal formulations was diluted compared to the oral formulations, based on the observation that the nasal mucosa is more sensitive to hyperosmolar solutions compared to the oral mucosa.

[0186] method The formulations described in Table 3 were prepared and tested. [Table 3]

[0187] 50 μl of the solution was sprayed into the left and right nostrils of two healthy volunteers using a nasal spray.

[0188] Tolerability was determined immediately after nebulization using a 4-point scale. 0: unacceptable, 1: unpleasant, 2: acceptable, 3: comfortable.

[0189] Secretory stimulation was determined after 2 min using a three-point scale. 0: no secretion, 1: secretion, 2, runny nose.

[0190] The nasal spray was repeated twice, with tolerability and secretory irritation noted after each spray.

[0191] Results and Conclusions The average results are shown in Table 4. Both formulations 6 and CX have a glycerol concentration of 54% by weight, which when sprayed into the nasal cavity causes severe stinging and severe runny nose. Reducing the glycerol concentration to 30% by weight and adding xylitol at increasing concentrations from 5 to 25% by weight (formulations 1 to 5) completely eliminated the stinging sensation and tolerability was experienced as acceptable to pleasant. Furthermore, slight secretory irritation was observed with formulations 4 and 5.

[0192] These findings indicate that it is possible to increase the xylitol concentration to 25% by weight in a 30% by weight glycerol solution without increasing the stinging sensation when such a solution is sprayed into the nasal cavity, supporting the suitability of glycerol / xylitol-based formulations for delivery of trypsin to the nasal cavity since trypsin is stabilized in such solutions (Example 2). [Table 4]

[0193] Example 5. Virucidal effect of throat spray and nasal spray formulations The aim of the study was to determine the virucidal effect of a throat spray formulation (for oral use) and a nasal spray formulation against the SARS-CoV-2 virus. In contrast to the throat spray, the nasal spray targets the nasal cavity, which is the primary site of viral upper respiratory tract infection.

[0194] method The study was designed to measure the virucidal efficacy of the disclosed formulations, thereby determining the potential of the formulations to inactivate the target virus-SARS-CoV-2 in suspension. The study followed the ASTM International test method entitled E1052 "Standard Test Method for Evaluating the Activity of Microbicides Against Viruses in Suspension". Severe Acute Respiratory Syndrome Associated Coronavirus 2 (SARS-CoV-2, COVID-19 virus), strain: USA-WA1 / 2020, source: BEI Resources, NR-52281 was used. Virus stocks were stored at ultra-low temperatures prior to use and thawed on the day of testing. Challenge virus stocks contained 5.0% serum. The compositions of the formulations tested were prepared as described in Table 5. The test formulations were pre-equilibrated to the test temperature prior to testing. Test solutions were evaluated in one replicate and with one contact time. For each experiment, an aliquot of 0.3 mL of virus stock was added to a mixture of 1.4 mL of test solution + 1.3 mL of phosphate buffer and mixed by vortexing. The mixture was incubated at 35-37 °C for 30 min. After incubation, an aliquot of the reaction mixture or the entire reaction mixture was removed and immediately mixed with an equal volume of newborn calf serum (NCS) and then vortexed. Selected dilutions were inoculated into host cells to assay the amount of infectious viral units.

[0195] Control Experiment Multiple controls were performed, including a virus recovery control, a neutralizer efficacy / virus interference control, a cytotoxicity control, a media negative control, and a virus stock titer control. The neutralizer efficacy / virus interference control was performed to determine whether there was any residual active component after neutralization and whether the neutralized test article interfered with virus infectivity. All controls were performed simultaneously with the test, incubated under the same conditions, and assayed in the same manner as the test.

[0196] i. Virus recovery control experiment: To a 2.7 mL aliquot of Minimum Essential Medium (MEM) + 2% NCS, 0.3 mL of virus was added and mixed by vortexing. Thirty minutes after virus addition (contact time), an aliquot of the reaction mixture or the entire reaction mixture was mixed with an equal volume of NCS by vortexing. Selected dilutions were inoculated into host cells to assay the amount of infectious virus as described in the "Infectivity Assay" section. The results of this control experiment were used as the input virus amount and compared to the results of the test article to evaluate the reduction of virus by the test article.

[0197] ii. Neutralizer Efficacy / Virus Interference Control Experiments: This control experiment determined whether there was any residual active component after neutralization and whether the neutralized test article interfered with virus infectivity. 0.3 mL of Minimum Essential Medium (MEM) + 2% NCS was added to a 2.7 mL aliquot of test article (1.4 mL test solution + 1.3 mL phosphate buffer), mixed by vortexing, and held for the contact time. After the contact time was over, an aliquot of the reaction mixture or the entire reaction mixture was immediately mixed with an equal volume of Minimum Essential Medium (MEM) + 2% NCS medium by vortexing and divided into two parts for the cytotoxicity control experiment and the neutralizer efficacy / virus interference control experiment, and processed as tests.

[0198] iii. Cytotoxicity control experiments: Selected dilutions of samples obtained from the neutralizer efficacy / virus interference control test setup were inoculated onto host cells and incubated with the samples as described for the test procedure. The status of the host cells was recorded at the end of the incubation period. Cytotoxic effects should be distinguished from virus-specific cytopathic effects, which will be evident in cultures of stock titers and virus recovery controls.

[0199] iv. Cell viability control experiment: During the incubation phase of the test, at least four wells were inoculated with the appropriate medium. This control experiment demonstrates whether the cells are viable throughout the assay period. In addition, it verifies the sterility of the medium used throughout the assay period.

[0200] v. Viral stock titer control experiment: Aliquots of the virus stocks used in the study were directly serially diluted and inoculated into host cells to confirm the titer of the stock virus. This control experiment demonstrates whether the titer of the stock virus is appropriate for use and whether the virus infectivity assay can be performed properly.

[0201] Infectivity assay: Residual infectious virus in test articles and controls was detected by virus-induced cytopathic effect (CPE).

[0202] Selected dilutions of the mixture to be tested were added to the cultured cell monolayers with at least 4 wells per dilution per sample. The inoculated plates were incubated at 4 °C for 2 h at 5 ± 3% CO 2 The cells were incubated at 36 ± 2 °C for 4 to 9 days in 100 mL of PBS. Prior to inoculation, the host cells were washed twice with phosphate-buffered saline. The host cell cultures were observed and, if necessary, refed during the incubation period. After completion of the incubation period, the host cells were examined for the presence of infectious virus. The resulting virus-specific CPE and test article-specific cytotoxic effects, if present, were scored by examining both the test article and the control. Virus was detected by staining with virus-specific antibodies, if necessary.

[0203] Calculation: The 50% tissue culture infectious dose per mL (TCID50 / mL) was determined using Spearman-Karber (Karber G. et al., 1931) or other appropriate methods such as Reed and Muench (Reed and Muench, 1938).

[0204] The Log10 reduction factor (LRF) was calculated in the following way: Log10 reduction factor = virus recovery control (Log10 TCID50) - test substance (Log10 TCID50) During the ceremony: Viral load (Log10 TCID50) = viral titer (Log10 TCID50 / mL) + Log10 [volume (mL) x volume correction (e.g., neutralization)].

[0205] Results and Conclusions The formulations of TS01 and TS02 are shown in Table 5. Only the nasal spray contains carrageenan. The virucidal efficacy of the throat spray (TS01) and the nasal spray (TS02) is shown in Table 6. [Table 5]

[0206] Virus stock titer controls for each assay confirmed that appropriate titers were used in the experiments and that sufficient virus was recovered for the virus recovery controls. No cytotoxicity was detected at any dilution. All controls met the criteria for a valid test.

[0207] Data from the SARS-CoV-2 virus reduction studies are shown in Table 6. The data demonstrate that both the nasal spray and throat spray formulations reduce SARS-CoV-2 virus by >99.9% in vitro.

[0208] This indicates that targeting the nose and mouth with antibacterial sprays, such as nasal sprays or mouth sprays, respectively, is effective in preventing or reducing the chances of contracting the virus. [Table 6]

[0209] Example 6. Rheological Experiments The purpose of this experiment was to determine how the viscosity changed upon the addition of carrageenan.

[0210] method Different formulations containing glycerol (55% by weight) and carrageenan (0-1% by weight) were mixed and the instantaneous viscosity of the solution was measured. A throat spray solution, TS03 (TS01 + 0.05% by weight carrageenan) was also evaluated.

[0211] Rheological measurements - rotational rheometer: The rheological properties of the formulations were investigated at high shear rates using a continuous shear technique and were performed using a controlled stress Malvern Rheometer (Malvern Instruments) with a plate of 20 mm geometric radius. The frequency sweep method was performed from 0.1 Hz to 10 Hz with a shear strain of 0.8% at 25 °C, and the shear rate method table shows the shear rate at 25 °C and 0.1 s -1 From 100s -1 The shear stress was measured by this method, and the apparent viscosity was calculated by dividing the shear stress by the shear rate.

[0212] Results and Conclusions The results are shown in Figure 5. It was possible to add at least 0.1 wt% carrageenan without affecting the viscosity. The addition of carrageenan to TS01 (to give TS03) did not significantly increase the viscosity, so the formulation can be used for spraying. The observed increase in viscosity suggests an increase in mucoadhesion.

[0213] Example 7. Virucidal effect of throat spray against influenza A virus The objective of the study was to determine the virucidal effect of the throat spray formulation against influenza A virus strain H3N2.

[0214] Materials and Methods This test had the same experimental setup as fully described in Example 5. This test also followed the ASTM International test method called E1052 "Standard Test Method for Evaluating the Activity of Microbicides Against Viruses in Suspension". The following specific modifications were made to the experimental setup of Example 5. Virus tested: Influenza A virus strain, H3N2, Charles River Laboratories. Host cell line: MDCK cells, ATCC CCL-34. Dilution medium: Minimum Essential Medium (MEM) + 1.0 μg / ml trypsin. Neutralizer: MEM + 1% fetal bovine serum (FBS). Incubation period 6 days.

[0215] Results and Conclusions The throat spray solutions (Table 5) were tested for their ability to inactivate influenza A virus (H3N2) exposed to TS01 throat spray solution in suspension for 30 minutes at 36°C. Assay virus stock titer controls confirmed that appropriate titers were used in the experiments and that sufficient virus was recovered for the virus recovery controls. No cytotoxicity was detected at any dilution. All controls met the criteria for a valid test.

[0216] The viral reduction with TS01 solution against influenza A virus (H3N2) was 1.25 log reduction, corresponding to a reduction of 94.4%. TS01 solution is also effective against influenza viruses in vitro.

[0217] Example 8. Inhibition of the cytopathic effect of respiratory syncytial virus (RSV) To evaluate the antiviral activity against RSV, an inhibition assay based on cytopathic effect (CPE) in HEp-2 cells was used for T01 and T02 (Table 5). CPE is a change in cell morphology caused by cytopathogenic virus infection.

[0218] Materials and Methods CPE inhibition assay On day -1, HEp-2 cells were seeded in clear 96-well plates at 1.30E+04 cells per well. T01 and T02 were diluted 1:10 in PBS and added to HEp-2 cells in triplicate. Cells were incubated at 37°C for 1 hour. Approximately 200 PFU / well of RSV A2 (0.02 MOI) was then added to the TA / cell wells. Virus only and cells only were also included for control experiments and calculations, and an internal assay control was also included for assay validation. HEp-2 cells were incubated at 37°C for 3 days. On day 3 post-infection, cells were immunostained, optical density was read, and the percentage of viral inhibition was determined.

[0219] Cytotoxicity assay T01 and T02 were diluted 1:10 in PBS and incubated with HEp-2 cells seeded in black 96-well plates for 3 days. Wells with cells only and media only were added. After 3 days, cells were lysed to assess ATP content using Promega's Cell Titer Glo kit. Luciferase luminescence was read in relative light units (RLU) to determine percent cytotoxicity.

[0220] Results and Conclusions Both T01 and T02 showed inhibition against RSV, but both solutions also showed high toxicity to HEp-2 cells (Table 7). High glycerol concentrations are known to be generally cytotoxic to human cells due to their high osmolality, therefore, at least for the T01 formulation, further dilutions can be performed to verify inhibition. [Table 7]

[0221] Example 9. In vitro antiviral effect against SARS-CoV-2 (Omicron variant) For this assay, the vesicular stomatitis virus (VSV) glycoprotein (G) was replaced with the SARS-CoV-2 B.1.1.529 (Omicron) spike (S) protein to generate a recombinant virus that can be safely handled at biosafety level 2. In this example, T01 and T02 were incubated with cells, and then virus was added to infect the cells. In Example 5, virus and T01 or T02 were pre-incubated and then added to the cells.

[0222] Materials and Methods Luciferase-based inhibition assay On day -1, Vero cells were seeded in black 96-well plates at 5.00E+04 cells per well. T01 and T02 were diluted 1:1 and 1:10 in PBS and added to Vero cells in triplicate. Cells were incubated at 37°C for 1 hour. Approximately 10,000 RLU of rVSV-SARS-CoV-2 B.1.1.529 Omicron was then added to the TA / cell wells. Virus only and cells only were also included for control experiments and calculations, and an internal assay control was also included for assay validation. Vero cells were incubated at 37°C for 24 hours. Firefly luciferase activity was detected using the Bright-Glo™ Assay System kit (Promega) to determine the percentage of viral inhibition.

[0223] Cytotoxicity assay T01 and T02 were diluted 1:1 and 1:10 in PBS and incubated with Vero cells seeded in black 96-well plates for 24 hours. Wells with cells only and media only were added. After 24 hours, cells were lysed to assess ATP content using Promega's Cell Titer Glo kit. Luciferase luminescence was read in relative light units (RLU) to determine percent cytotoxicity.

[0224] Results and Conclusions Low cytotoxicity was observed for both T01 and T02 at 1:10 dilution. Both T01 and T02 showed inhibition against SARS-CoV-2B.1.1.529 at 1:10 diluted formulations. High cytotoxicity was observed for both T01 and T02 at 1:1 dilution. Results are shown in Table 8. [Table 8]

[0225] Example 10: Stability of nasal spray and throat spray formulations The purpose of the experiment was to measure the enzyme stability in TS01 and TS02.

[0226] method TS01, TS02, TS02 without xylitol, and commercial trypsin-containing oral spray (CX) were stored in sealed glass bottles. The stability of trypsin in each formulation composition was tested by placing it under stress conditions (40±2°C / 100% RH) for up to 97 days. Enzyme activity was measured using the same method as described in Example 1.

[0227] Results and Conclusions The results are shown in Figure 6. TS01 showed the highest stability. Both TS01 and TS02 are more stable than CX. TS02 without xylitol is not stable.

[0228] References Allan GM,Arroll B.Prevention and treatment of the common cold:making sense of the evidence.CMAJ.Can Med Assoc;2014;186(3):190-9. Craik CS et al.Proteases as therapeutics.Biochem J.2011;435:1-16. D'Souza et al., 2015 Gudmundsdottir, Agusta, et al. Inactivation of SARS-CoV-2 and HCoV-229E in vitro by ColdZyme® a medical device mouth spray against the common cold. J Med Virol. 2021;93:1792-1795. Harkema JR, Carey SA, Wagner JG. The nose revisited: a brief review of the comparative structure, function, and toxicologic pathology of the nasal epithelium. Toxicologic pathology. Sage Publications;2006;34(3):252-69. Heikkinen T, Jarvinen A. The common cold. Lancet. 2003 Jan 3;361(9351):51-9. Jackson GG., Dowling HF, Spiesman IG, Boand AV.. Transmission of the common cold to volunteers under controlled conditions. I. The common cold as a clinical entity. AMA Arch Intern Med. 1958 Feb;101(2):267-78. Johnston SL et al. The relationship between upper respiratory infections and hospital admissions for asthma: a time-trend analysis. Am J Respir Crit Care Med. American Public Health Association;1996;154(3):654-60. Karber,G.Beitrag zur kollecktiven Behandlung pharmakologischer Reihenversuche.Arch.Exptl.Pathol.Pharmakol,162,480-483,1931. Lenoir J,Adriaens E,Remon JP.New aspects of the Slug Mucosal Irritation assay:predicting nasal stinging,itching and burning sensations.Journal of Applied Toxicology.Wiley Online Library;2011;31(7):640-8. Mallia,Patrick,and Sebastian L.Johnston.Influenza infection and COPD.International journal of chronic obstructive pulmonary disease;2007:2(1):55-64. Meena et al,A Review on Microbial Pathogenesis and Host Response.Model Organisms for Microbial Pathogenesis,Biofilm Formation and Antimicrobial Drug Discovery.Springer,Singapore,2020.47-60. Pappas DE,Hendley JO,Hayden FG,Winther B.Symptom profile of common colds in school-aged children.The Pediatric infectious disease journal.LWW;2008;27(1):8-11. Posch W,Vosper J,Zaderer V,Noureen A,Constant S,Bellmann-Weiler R,et al.ColdZyme Maintains Integrity in SARS-CoV-2-Infected Airway Epithelia.Mbio.Am Soc Microbiol;2021;12(2). Ramalingam S,Graham C,Dove J,Morrice L,Sheikh A.A pilot,open labelled,randomised controlled trial of hypertonic saline nasal irrigation and gargling for the common cold.Scientific reports.Nature Publishing Group;2019;9(1):1-11. Reed L.J and Muench H.A simple method of estimating fifty percent endpoints.American Journal of Epidemiology,Volume 27,Issue 3,May 1938,Pages 493-497. Salade L,Wauthoz N,Goole J,Amighi K.How to characterize a nasal product.The state of the art of in vitro and ex vivo specific methods.Int J Pharm.Elsevier;2019;561:47-65. Sungnak W,Huang N,Becavin C,Berg M,Queen R,Litvinukova M,et al.SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells together with innate immune genes.Nature medicine.Nature Publishing Group;2020;26(5):681-7. Trangsrud AJ,Whitaker AL,Small RE.Intranasal corticosteroids for allergic rhinitis.Pharmacotherapy:The Journal of Human Pharmacology and Drug Therapy.Wiley Online Library;2002;22(11):1458-67. Zelikin AN,Stellacci F.Broad-Spectrum Antiviral Agents Based on Multivalent Inhibitors of Viral Infectivity.Advanced Healthcare Materials.Wiley Online Library;2021;10(6):2001433.

[0229] item 1. A composition comprising: i. 0.005 to 1.0% w / w of a peptidase, and ii. The composition comprising 1.0-70% w / w sugar alcohol.

[0230] 2. The composition according to item 1, i. 0.005-1.0% w / w of a peptidase; ii. 20-70% w / w glycerol, and iii. The composition comprising 1.0-65% w / w xylitol.

[0231] 3. A composition according to any one of the preceding items, wherein the composition comprises 0.005-1.0% w / w of peptidase, such as 0.01-0.075% w / w of peptidase, for example 0.015-0.055% w / w of peptidase.

[0232] 4. The composition of any one of the preceding claims, wherein the peptidase is selected from the group consisting of serine proteases, threonine proteases, cysteine ​​proteases, aspartic acid proteases, glutamic acid proteases and metalloproteases, functional homologues thereof, or functionalized derivatives thereof.

[0233] 5. The composition of any one of the preceding items, wherein the peptidase is trypsin, a functional homologue thereof, or a functionalized derivative thereof.

[0234] 6. The composition of any one of the preceding items, wherein the peptidase is trypsin.

[0235] 7. The composition according to any one of the preceding items, wherein the composition comprises 0.005-1.0% w / w trypsin, such as 0.01-0.075% w / w trypsin, for example 0.015-0.055% w / w trypsin.

[0236] 8. The composition according to any one of the preceding items, i. 0.005-1.0% w / w trypsin; ii. 20-70% w / w glycerol, and iii. The composition comprising 1.0-65% w / w xylitol.

[0237] 9. The composition of any one of the preceding items, wherein the composition comprises 0.005-1.0% w / w trypsin, such as 0.01-0.075% w / w trypsin, such as 0.015-0.055% w / w trypsin.

[0238] 10. The composition according to any one of the preceding items, wherein the composition further comprises 0.003 to 0.2% w / w of a salt comprising a divalent cation, such as 0.003 to 0.15% w / w, for example 0.003 to 0.1% w / w of a salt comprising a divalent cation.

[0239] 11. The composition according to any one of the preceding items, wherein the peptidase is trypsin, a functional homologue thereof, or a functionalized derivative thereof, and the composition further comprises 0.003 to 0.2% w / w of a salt comprising a divalent cation, such as 0.003 to 0.15% w / w, for example 0.003 to 0.1% w / w of a salt comprising a divalent cation.

[0240] 12. A composition according to any one of the preceding items, wherein the composition comprises 35-70% w / w sugar alcohol, such as 40-65% w / w sugar alcohol, for example 45-60% w / w sugar alcohol.

[0241] 13. The composition of any one of the preceding items, wherein the sugar alcohol is glycerol, erythritol, xylitol, mannitol, arabitol, maltitol, lactitol, isomalt, sorbitol, hydrogenated starch hydrolysates or propylene glycol, or other related polyols.

[0242] 14. A composition according to any one of the preceding items, wherein the composition comprises 20-70% w / w glycerol, such as 25-65% w / w glycerol, for example 27-57% w / w glycerol.

[0243] 15. A composition according to any one of the preceding items, wherein the composition comprises 1.0-65% w / w xylitol, such as 3.0-50% w / w xylitol, for example 4.5-30% w / w xylitol.

[0244] 16. The composition according to any one of the preceding items, wherein the combined amount of glycerol and xylitol is at least 40% w / w, such as at least 45% w / w, such as at least 50% w / w.

[0245] 17. The composition of any one of the preceding items, wherein the composition further comprises a buffer.

[0246] 18. The composition according to any one of the preceding items, wherein the composition comprises 0.01-2.4% w / w of a buffer solution, such as 0.07-0.3% w / w of a buffer solution, for example 0.09-0.2% w / w of a buffer solution.

[0247] 19. The composition according to any one of the preceding items, wherein the concentration of the buffer is 1 to 200 mM, for example, 5 to 25 mM, for example, 8 to 15 mM.

[0248] 20. The composition of any one of the preceding items, wherein the buffer is selected from the group consisting of tris(hydroxymethyl)aminomethane (Tris), 3-(N-morpholino)propanesulfonic acid (MOPS) or phosphate.

[0249] 21. The composition according to any one of the preceding items, wherein the pH is between 5.5 and 8.5.

[0250] 22. The composition according to any one of the preceding items, wherein the pH is between 6.2 and 7.0, for example between 6.4 and 6.6.

[0251] 23. The composition according to any one of the preceding items, wherein the pH is from 7.2 to 8.5, for example from 7.4 to 7.6.

[0252] 24. The composition of any one of the preceding items, wherein the composition further comprises water.

[0253] 25. The composition according to any one of the preceding items, wherein the composition comprises at most 50% w / w water, such as at most 40% w / w, such as at most 30% w / w water.

[0254] 26. The composition according to any one of the preceding items, wherein the composition further comprises hyaluronic acid, for example, 0.0001-2.0% w / w hyaluronic acid, for example, 0.0001-1.5% w / w, for example, 0.0001-1.0% w / w, for example, 0.001-1.5% w / w, for example, 0.01-1.0% w / w, for example, 0.02-0.5% w / w, for example, 0.0001-0.1% w / w, for example, 0.0001-0.01% hyaluronic acid.

[0255] 27. The composition of any one of the preceding items, wherein the composition further comprises a divalent cation.

[0256] 28. The composition of any one of the preceding items, wherein the divalent cation is calcium.

[0257] 29. The composition of any one of the preceding items, wherein the salt containing a divalent cation is a pharma- ceutically acceptable salt of calcium, including hydrates thereof.

[0258] 30. A composition according to any one of the preceding items, wherein the composition further comprises 0.003-0.2% w / w of a calcium salt, such as 0.003-0.15% w / w of a calcium salt, for example 0.003-0.1% w / w of a calcium salt.

[0259] 31. A composition according to any one of the preceding items, wherein the composition further comprises calcium ions, for example, 0.002-0.2% w / w calcium ions, for example, 0.003-0.15% w / w calcium ions, for example, 0.003-0.1% w / w calcium ions.

[0260] 32. The salt containing a divalent cation is CaCl 2 Item 11. The composition of any one of the preceding items, wherein

[0261] 33. The composition of any one of the preceding items, wherein the salt containing a divalent cation is a pharma- ceutically acceptable inorganic salt of calcium, including hydrates thereof.

[0262] 34. A composition according to any one of the preceding items, wherein the composition comprises 0.003-0.2% w / w of divalent calcium, such as 0.003-0.15% w / w, for example 0.003-0.1% divalent calcium.

[0263] 35. A composition according to any one of the preceding items, wherein the composition comprises 0.002-0.2% w / w calcium chloride, such as 0.005-0.02% w / w calcium chloride, for example 0.010-0.015% w / w calcium chloride.

[0264] 36. The composition, i. 0.005-1.0% w / w trypsin; ii. 20-70% w / w glycerol; iii. 1.0-65% w / w xylitol; iv. 0.0001-1.5% w / w hyaluronic acid; v. 0.002 to 0.2% w / w CaCl 2 , and vi. A composition according to any one of the preceding items, comprising or consisting essentially of 0.01-1.2% w / w of a buffer solution.

[0265] 37. The composition, i. 0.015-0.055% w / w trypsin; ii. 27-57% w / w glycerol; iii. 4.5-30% w / w xylitol; iv. 0.0002-0.02% w / w hyaluronic acid; v. 0.010-0.015% w / w CaCl 2 , and vi. A composition according to any one of the preceding items, comprising or consisting essentially of 0.1-0.14% w / w buffer.

[0266] 38. The composition of any one of the preceding items, further comprising carrageenan.

[0267] 39. A composition according to any one of the preceding items, wherein the composition comprises 1% w / w or less of carrageenan, such as 0.5% w / w or less of carrageenan, for example 0.1% w / w or less of carrageenan.

[0268] 40. The composition of any one of the preceding items, further comprising a flavoring agent.

[0269] 41. The composition of any one of the preceding items, wherein the flavoring agent is natural or non-natural.

[0270] 42. The composition of any one of the preceding items, wherein the flavoring agent is spearmint or eucalyptus.

[0271] 43. A composition according to any one of the preceding items, wherein the composition comprises 0.01 to 0.4% w / w of flavouring agent, for example 0.02 to 0.1% w / w of flavouring agent, for example 0.035 to 0.055% w / w of flavouring compound.

[0272] 44. The composition, i. 0.02-0.05% w / w of a peptidase; ii. 29-56% w / w glycerol; iii. 4 to 25% w / w xylitol; iv. 0.0002-0.0004% w / w hyaluronic acid; v. 0.01-0.02% w / w CaCl 2 , vi. 0.1-0.2% w / w buffer solution; vii. 0-0.1% w / w carrageenan, and viii. A composition according to any one of the preceding items, comprising or consisting essentially of 0-0.05% w / w flavouring agent.

[0273] 45. The composition of any one of the preceding items, wherein the composition is delivered in the form of a spray, lozenge, troche, chewing gum, gel, or liquid.

[0274] 46. ​​The composition of any one of the preceding items, wherein the composition is a sprayable composition.

[0275] 47. The composition of any one of the preceding items, wherein the composition is in the form of a spray.

[0276] 48. The composition of any one of the preceding items, wherein the spray is a nasal spray or a throat spray.

[0277] 49. The composition according to any one of the preceding items, having a viscosity of at most 0.015 Pa.s, for example at most 0.015 Pa.s, as assessed using a controlled stress Malvern Rheometer (Malvern Instruments) with a plate of geometric radius 20 mm at 25°C.

[0278] 50. A composition according to any one of the preceding items for use in medicine.

[0279] 51. A composition according to any one of the preceding items for use in the prevention and / or treatment in and / or for a mammal of a disease, disorder or condition selected from the group consisting of microbial infections and oral diseases such as gingivitis or periodontitis.

[0280] 52. The composition for use according to any one of the preceding items, wherein the microbial infection is selected from the group consisting of a viral infection, a bacterial infection, a fungal infection, and a yeast infection.

[0281] 53. The composition for use according to any one of the preceding items, wherein the microbial infection is a viral infection.

[0282] 54. The composition for use according to any one of the preceding items, wherein the viral infection is a viral upper respiratory tract infection.

[0283] 55. A composition for use according to any one of the preceding items, wherein the viral upper respiratory tract infection results in a cold, influenza, rhinitis, sinusitis, COVID-19 such as the Omicron variant, or a disease caused by a viral upper respiratory tract infection.

[0284] 56. The composition for use according to any one of the preceding items, wherein the viral infection is due to a virus selected from the group consisting of rhinovirus, influenza virus such as influenza A virus, respiratory syncytial virus (RSV), coronavirus, parainfluenza virus, adenovirus, enterovirus, metapneumovirus, and other infectious viruses.

[0285] 57. The composition for use according to any one of the preceding items, wherein the viral infection causes an infectious disease such as cold, influenza, rhinitis, sinusitis, bronchitis, Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), coronavirus disease 2019 (COVID-19) such as COVID-19 Omicron variant, pneumonia, viral meningitis, herpangina, herpes virus, papilloma virus, or any other disease caused by a viral infection.

[0286] 58. A composition for use according to any one of the preceding items, wherein the prevention and / or treatment is the prevention and / or treatment of symptoms associated with a viral infection, such as sore throat, fatigue, nasal discharge, stuffy nose, headache, cough, sneezing, and / or fever.

[0287] 59. A composition for use according to any one of the preceding items, wherein the composition is nasal.

[0288] 60. A composition for use according to any one of the preceding items, wherein the composition is formulated for intranasal administration.

[0289] 61. A composition for use according to any one of the preceding items, wherein the composition is for oral use.

[0290] 62. Use of a composition according to any one of the preceding items in the manufacture of a medicament for the treatment and prevention of a disease, disorder or condition selected from the group consisting of microbial infection, skin disease, and oral disease.

[0291] 63. A method for the treatment and / or prevention of a disease, disorder, or condition selected from the group consisting of microbial infections, skin diseases, and oral diseases, comprising administering to an individual in need thereof a composition described in any one of the preceding items.

[0292] 64. A method for the prevention and / or reduction of viral infection, comprising administering a composition according to any one of the preceding items.

[0293] 65. A method for increasing microbial diversity, comprising administering a composition described in any one of the preceding items.

[0294] 66. A method for producing a composition according to any one of the preceding items, comprising: i. peptidases, and ii. mixing a sugar alcohol.

[0295] 67. A method for producing a composition according to any one of the preceding items, comprising: i. trypsin, ii. glycerol, and iii. The method comprising mixing xylitol.

[0296] 68. A method for producing a composition according to any one of the preceding items, comprising: i. glycerol, ii. xylitol, iii.Buffer, iv. hyaluronic acid, and v. The above method, comprising mixing a peptidase.

[0297] 69. A method according to any one of the preceding items, comprising: i. carrageenan, and / or ii. The method further comprising mixing a flavoring agent.

Claims

1. 1. A composition comprising: i. 0.005-1.0% w / w trypsin; ii. 20-70% w / w glycerol, and iii. The composition comprising 1.0 to 65% w / w of xylitol.

2. The composition comprises: i. 0.01-0.075% w / w trypsin; ii. 25-65% w / w glycerol, or iii. The composition of claim 1, comprising 3.0-50% w / w xylitol.

3. 10. The composition of claim 1, wherein the composition further comprises 0.003 to 0.2% w / w of a salt comprising a divalent cation.

4. 2. The composition of claim 1, wherein the composition comprises 35-70% w / w of a sugar alcohol, wherein the sugar alcohol is glycerol, erythritol, xylitol, mannitol, arabitol, maltitol, lactitol, isomalt, sorbitol, hydrogenated starch hydrolysate or propylene glycol, or other related polyol.

5. 2. The composition of claim 1, wherein the combined amount of glycerol and xylitol is at least 40% w / w.

6. The composition comprises: i. buffer solution, ii. water, iii. hyaluronic acid, or iv. carrageenan, The composition of claim 1 further comprising:

7. The composition comprises: i. 1-200 mM buffer; ii. at most 50% w / w water; iii. 0.0001-2.0% w / w hyaluronic acid, or iv. at most 1% w / w carrageenan; The composition of claim 6 comprising:

8. 7. The composition of claim 6, wherein the buffer is tris(hydroxymethyl)aminomethane (Tris), 3-(N-morpholino)propanesulfonic acid (MOPS), or phosphate.

9. The composition described in claim 1, wherein the pH of the composition is 5.5 to 8.

5.

10. The composition of claim 3 , wherein the salt containing a divalent cation is a pharmaceutically acceptable salt of calcium, including hydrates thereof.

11. The salt containing a divalent cation is CaCl ２ The composition of claim 3, wherein

12. The composition comprises: i. 0.005-1.0% w / w trypsin; ii. 20-70% w / w glycerol; iii. 1.0 to 65% w / w of xylitol; iv. 0.0001-1.5% w / w hyaluronic acid; v. 0.002-0.2% w / w CaCl ２ , and vi. The composition of claim 1, comprising or consisting essentially of 0.01-1.2% w / w of a buffer.

13. The composition comprises: i. 0.015-0.055% w / w trypsin; ii. 27-57% w / w glycerol; iii. 4.5-30% w / w xylitol; iv. 0.0002-0.02% w / w hyaluronic acid; v. 0.010-0.015% w / w CaCl ２ , and vi. The composition of claim 1, comprising or consisting essentially of 0.1-0.14% w / w of a buffer.

14. The composition comprises: i. 0.02-0.05% w / w trypsin; ii. 29-56% w / w glycerol; iii. 4-25% w / w xylitol; iv. 0.0002-0.0004% w / w hyaluronic acid; v. 0.01-0.02% w / w CaCl ２ , vi. 0.1-0.2% w / w of a buffer solution; vii. 0-0.1% w / w of carrageenan, and viii. The composition of claim 1, comprising or consisting essentially of 0-0.05% w / w flavoring agent.

15. 10. The composition of claim 1, wherein the composition is in the form of a spray, lozenge, troche, chewing gum, gel, or liquid.

16. The composition of claim 1 , wherein the spray is a nasal spray or a throat spray.

17. A pharmaceutical composition comprising the composition of claim 1 for use in the prevention or treatment of a disease, disorder, or condition selected from the group consisting of microbial infections and oral diseases in and / or for a mammal.

18. The microbial infection is selected from the group consisting of a viral infection, a bacterial infection, a fungal infection, and a yeast infection, or ii. The oral disease is selected from gingivitis and periodontal disease; 18. The pharmaceutical composition of claim 17.

19. 19. The pharmaceutical composition of claim 18, wherein the viral infection is a viral upper respiratory tract infection.

20. The viral infection is i. Caused by a virus selected from the group consisting of rhinovirus, influenza virus, respiratory syncytial virus (RSV), coronavirus, parainfluenza virus, adenovirus, enterovirus, metapneumovirus, and other infectious viruses; or ii. Causes an infectious disease selected from colds, influenza, rhinitis, sinusitis, bronchitis, severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), coronavirus disease 2019 (COVID-19) such as COVID-19 Omicron variant, pneumonia, viral meningitis, herpangina, herpesvirus, papillomavirus, or any other disease caused by a viral infection; 19. The pharmaceutical composition of claim 18.

21. 18. The pharmaceutical composition according to claim 17, wherein the prevention or treatment is the prevention or treatment of one or more symptoms selected from sore throat, fatigue, rhinorrhea, nasal congestion, headache, cough, sneezing, and fever.

22. A method for producing the composition of claim 1, comprising: i. trypsin, ii. glycerol, and iii. The method, further comprising mixing xylitol.

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