WO2016067283A1 - Virucidal compositions comprising tannins for nasal and pulmonary administration - Google Patents

Virucidal compositions comprising tannins for nasal and pulmonary administration Download PDF

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Publication number
WO2016067283A1
WO2016067283A1 PCT/IL2015/051048 IL2015051048W WO2016067283A1 WO 2016067283 A1 WO2016067283 A1 WO 2016067283A1 IL 2015051048 W IL2015051048 W IL 2015051048W WO 2016067283 A1 WO2016067283 A1 WO 2016067283A1
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composition
tannins
nasal
dry powder
water
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PCT/IL2015/051048
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French (fr)
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Howard RICE
Ian Solomon
Dr. Andreas BILSTEIN
Olaf SCHERNER
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Rice Howard
Ian Solomon
Bilstein Dr Andreas
Scherner Olaf
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Publication of WO2016067283A1 publication Critical patent/WO2016067283A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0043—Nose
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/12—Carboxylic acids; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/007—Pulmonary tract; Aromatherapy
    • A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
    • A61K9/0078—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a nebulizer such as a jet nebulizer, ultrasonic nebulizer, e.g. in the form of aqueous drug solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/02—Nasal agents, e.g. decongestants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses

Definitions

  • tannins are virucidal compounds in the sense that they attack the membrane wall of the virus.
  • the literature uses the terms antiviral and virucidal interchangeably, the more correct term for the activity of tannins is "virucidal" as the mode of action is one of destroying viruses on contact. While the virucidal properties of tannins in the form of tannic acid and tannin-containing solutions are well known, a 1953 paper was the first to explicitly describe the way in which influenza viruses are deactivated.
  • Influenza hemagglutinin (HA) is a glycoprotein found on the surface of the influenza virus, and thus we see that the mode of deactivation of influenza viruses by tannins is the denaturing of the proteins on contact with the external membrane of the virus.
  • Liu and his co-workers Liu S, Li H, Chen L, Yang L, Li L, Tao Y, et al., "Epigallocatechin-3- gallate inhibition of Epstein-Barr virus spontaneous lytic infection involves ERK1/2 and PI3-K/Akt signaling in EBV-positive cells", Carcinogenesis 2013; 34: 627-37
  • BALB/c mice On the 3rd day of the treatment, the mice were infected with influenza A virus.
  • EGCG at 40 mg/kg per day dramatically improved the survival rate of the experimental animals, and also decreased virus replication and the mitigation of viral pneumonia in the lungs.
  • tannins are known to have antibacterial properties.
  • An overview of both the antiviral and antimicrobial effects of tannins can be found in the article: "Antimicrobial and antiviral activity of hydrolysable tannins", Mini Rev Med Chem. 2008 Oct; 8(12): 1179-87, by Buzzini P. et al.
  • the antimicrobial mechanism of tannins is thought to result from the astringent property of the tannin inducing complexation with enzymes or substrates on the membranes of the microorganisms (J. Antimicrob. Chemother. (2001) 48 (4): 487-491).
  • the role of tannins in combatting S. Aureus and P. aeruginosa have been shown in H.Akiyama et al.
  • tannins are knows to be astringent and, at effective concentrations, convey a stinging sensation on the tissue, where they are applied.
  • Natural tannin is an acidic compound, and the pH of a concentrated aqueous solution of tannic acid is lower than 3.5.
  • Aqueous acidic solutions cause a stinging sensation on mucosa and should be basified to pH>5 in order to be comfortable.
  • the use of tannin preparations within the nasal cavity - for example as a nasal spray or nasal drops - is problematic.
  • the other problem with liquid tannin formulations is their lack of stability due to hydrolysis.
  • WO 2011/082835 Al it is stated that "hydrosoluble tannins are not suitable for topical applications" .
  • the objective of the present invention is to overcome the formulation problems associated with delivering tannins such that the practical implementation of tannin-based compositions in the form of aerosols for nasal and pulmonary delivery will be enabled.
  • the present invention therefore comprises a comfortable, non-stinging aqueous formulation of tannins.
  • This comfortable composition results from dissolving the tannins in a mostly aqueous solution together with one or more compatible soothing agents and, optionally, also one or more suitable stabilizing agents.
  • the soothing agents that may be used for this purpose may be selected from the groups including osmolytes and demulcents (sometimes referred to as mucoprotective agents), but only where the specific soothing agent chosen does not react with tannic acid to form insoluble tannates.
  • said soothing agent will also be permitted for use in a nasal and/or pulmonary aerosol.
  • any stabilizing agent(s) used need to be compatible with tannic acid and allowed in a nasal and/or pulmonary aerosol.
  • a soothing agent or demulcent serves to form a soothing, protective film or complex when administered onto a mucous membrane surface, it serves to counteract and thus allay the stinging or astringent effect of the chemical agent dissolved within it.
  • compositions are provided for the nasal administration of tannin-containing aerosols and for the pulmonary administration of tannin-containing aerosols, including inhalation solutions.
  • Said compositions may be used for the purpose of combating viral infection in general, and influenza, rhinovirus, RSV and viral pneumonia specifically.
  • tannins are also known for their antibacterial characteristics, the use of the composition of the present invention will also cause decolonization of bacterial activity in the nasal cavity, lungs and sinuses. This is expected to reduce the risk of bacterial complications following an influenza or other viral infection and minimize the symptoms of diseases such as cystic fibrosis.
  • Figure 1 a shows the cytotoxicity of the pure 2% Ectoine solution.
  • Figure lb shows the cytotoxicity of the combination with 0.2% tannic acid.
  • Figure 2a shows the cytotoxicity of the pure 2% Ectoine solution on RLE cells.
  • Figure 2b shows the cytotoxicity of the combination with 0.2% tannic acid on RLE cells.
  • Figures 3a, 3b and 3c show the virucidal efficacy of the exemplary composition against influenza viruses A/CA/07/09 (H1N1), A/HK/1/68 (H3N2), and B/Lee/40 respectively.
  • Figure 4 shows a plot-graph of the cytotoxicity information from the nasal epithelial model on the same graph as data showing the virucidal efficacy against the three influenza viruses mentioned in Figs 3a-c.
  • Tannic acid as powder (Merck, USP/EP) was dissolved in water, at a number of concentrations in the 0.1% to 1% range. Due to the astringent or stinging effect caused by the spraying of this tannic acid solution into the nose, a number of potential soothing agents were examined for their potential addition to the composition.
  • Demulcents are a broad class of substances, including the synthetic demulcents methyl cellulose, glycerin or propylene glycol, the natural demulcents pectin and honey, and active extracts from Aloe Vera and other natural sources. Obviously, compounds that are available off-the-shelf in pharmaceutical grade (USP) are preferred choices for the composition of the present invention. Unfortunately, in preliminary work performed by the authors, the preferred demulcents glycerin and propylene glycol were both shown to be irritants when used in a nasal spray; whereas methyl cellulose made the formulation too viscous to spray.
  • USP pharmaceutical grade
  • tannic acid forms precipitates with a wide range of organic and inorganic compounds as di- and trivalent metal ions, amines, amino acids, peptides, proteins and some other polymers.
  • Precipitating tannates remove the tannic acid from solution and thus reduces its bioavailability.
  • tannic acid is incompatible in solutions with many active ingredients and excipients.
  • Aloe Vera which is available in USP grade (Aloway Natural Health Products, Haymeadow Crescent Faerie Glen, Pretoria East, South Africa) was tested for its applicability.
  • This compound has proved to be an excellent soothing agent for nasal sprays in general; and indeed it is used for that purpose in the Otrimer Aloe Vera product from Novartis Consumer Health (Nyon, Switzerland) where it serves to prevent the stinging caused by a saline nasal spray.
  • adding it to our tannic acid solutions caused a tannate precipitate to emerge from the solutions, and thus Aloe Vera was shown by experiment to be incompatible with tannic acid in solution.
  • Osmolytes include sugar alcohols (myoinositol, mannitol, sorbitol), quaternary amines (such as taurine, choline, betaine, betaine- glycine) and Ectoines.
  • Further candidates include glutamate, aspartate or proline, phosphatidylcholine, phosphatidylinositol diglycerol phosphate, phosphorylcholine, glycerophosphorylcholines, and amino acids such as glutamine, glycine and alanine.
  • Toxicity was determined via neutral red assay, where absorption was measured at 540 nm and a high absorption represents a healthy cell.
  • an existing Ectoine nasal spray solution comprising just 2% Ectoine (labelled "SNS01") was also diluted in cell medium and measured as described above.
  • Figure la shows the in vitro cytotoxicity of a solution comprising just 2% Ectoine; this being a comfortable nasal spray product which is currently marketed for symptomatic relief of rhinitis.
  • Figure lb shows the in vitro cytotoxicity of an exemplary composition of the present invention comprising 0.2% tannic acid + 2% Ectoine. Note that the combined composition is slightly more cytotoxic than the pure Ectoine one, but is still within reasonable limits. The comfort level of the above exemplary composition was then verified by testing on ten human subjects who each received a spray of this solution into both nostrils and did not experience any discomfort.
  • This exemplary composition (0.2% tannic acid and 2% Ectoine) was then tested at successive dilutions in order to determine whether, over the tested concentration range, tannic acid performed as an effective virucidal compound.
  • the cell model used was the CPE-based EC50 assay using MDCK cells, where the composition was tested against the three influenza viruses: A/CA/07/09 (H1N1), A/HK/1/68 (H3N2), and B/Lee/40.
  • the assay comprised the following steps: MDCK cells were seeded in 96-well plates and incubated overnight. The next day serial dilutions of the test article (starting at 25% final concentration, 2-fold dilutions) as well as a control compound were prepared in medium.
  • the growth medium was aspirated from the cells and the compound dilutions were added at 2x the final concentration for a one-hour incubation period. Afterwards the virus was added at an MOI of 0.01 and the cells were incubated for 6 days. The cells were then fixed and stained with crystal violet in glutaric dialdehyde solution. The optical density was determined and the EC50 was calculated using the uninfected (cells only) control as 0% CPE and the controls without compound (virus only) as 100% CPE using a 4-PL curve fit of the OD.
  • Figures 3a, 3b and 3c shows the virucidal efficacy of the exemplary composition against influenza viruses A/CA/07/09 (H1N1), A/HK/1/68 (H3N2), and B/Lee/40 respectively. Note that the composition exhibits efficient inhibition of all of these viruses at just a 25% concentration, and that the dose-response curves are similar to those of the standard antiviral drug Zanamivir (which is shown for reasons of comparison).
  • Figure 4 then plots the cytotoxicity information from the nasal epithelial model (above) on the same graph as data showing the virucidal efficacy against the three above influenza viruses (shown as a band between the two dashed lines). From this graph we a clear picture emerges of the therapeutic window of the composition of the present invention; at least for nasal delivery.
  • the two solid vertical lines define the concentrations at which the composition will be tolerable but nevertheless effective. Looking at this area, it is reasonable to state that the concentration of tannic acid should be between 0.02% and 0.2%. Additionally, there may well also be some room for the patient to tolerate some slight discomfort in return for greater efficacy, and therefore the upper concentration range of tannic acid could be raised to 2% in such cases. Note also that immediately after application, the composition will immediately be diluted with biological liquid (mucus) and thus the concentration delivered would be reduced significantly by this dilution; causing an initial, higher tannic acid concentration to rapidly be diluted towards the 0.2% level.
  • tannic acid is inherently unstable in aqueous solution. This instability results from both oxidation and hydrolysis of the tannic acid components. Accordingly, a number of approaches were used to address this stability problem: (1) reduction of the aqueous percentage of the composition by using a 75/25 water/propylene glycol mix instead of just water; (2) addition of one or more preservative agents, (3) addition of one or more antioxidants, (4) pH adjustment and (5) addition of one or more osmolytes; as detailed below:
  • composition of the present invention can be made "less aqueous" by using propylene glycol is, unfortunately, relatively limited as the propylene glycol itself was found to cause irritation of the nasal mucosa when used in concentrations above 25%.
  • preservative agents are potential candidates to increase the stability of the composition of the present invention. According to a review of preservative agents currently used in nasal sprays, "/ « vitro effects of preservatives in nasal sprays on human nasal epithelial cells” (Am J._Rhinol. 2008 Mar-Apr;22(2): 125-9. doi: 10.2500/ajr.2008.22.3154), of the two common preservatives, benzalkonium chloride and potassium sorbate, the former causes epithelial damage, and thus only the latter should be used. Our experiments also included propyl gallate, which is an antioxidant agent, often used as a preservative agent in foods, together with BHA and BHT.
  • propyl gallate should demonstrate a high degree of compatibility with tannic acid and thus preserve it relatively well.
  • the last is least preferred as it may iodize the tannic acid.
  • Antioxidant agents suitable for the composition of the present invention include ascorbic acid, tocapherol and glutathione.
  • preferred exemplary compositions according to the present invention contain from 0.02% to 2% by weight of tannins (for example tannic acid) and include at least one soothing agent.
  • tannins for example tannic acid
  • the concentration of tannin will be between 0.02% and 0.2%.
  • said compositions further comprise at least one antioxidant agent and at least one preservative agent. In some cases the antioxidant and the preservative may be the same agent.
  • the soothing agent may also serve as a preservative, in particular preserving against hydrolysis of the tannins.
  • composition of the present invention are as follows:
  • formulations may be made up in either a purely aqueous solution (i.e. 100% water) or, in a preferred embodiment, a water / propylene glycol mixture may be used.
  • a water / propylene glycol mixture may be used.
  • using the latter mixture was shown to reduce the rate of hydrolysis, thus enhancing the stability of the product.
  • the water / propylene glycol ratios used for such a mixture can range from 99%/l% to 75%/25%; as using above 25% propylene glycol makes the composition too viscous to spray well and somewhat painful to the nasal tissues.
  • some preferred formulations incorporating propylene glycol are those of composition no. 2 above, in a 75/25 mixture or an 80/20 mixture.
  • antioxidant agents and stabilizers can be added to the above compositions.
  • oil of mint, menthol, eucalyptus oil and/or saline or sea water may be added to the composition, to open the nasal passages.
  • aerosol refers to a colloid of fine solid particles or liquid droplets, in air or another gas.
  • the composition of the present invention may be formulated either for use as a liquid-droplet aerosol or as a fine -powder aerosol.
  • Liquid droplets may, for example, be delivered using a spray bottle for nasal administration, or by a nebulizer for pulmonary administration.
  • the liquid formulation of this composition can be provided to the patient in a number of forms: (a) in final liquid form, ready for aerolization; (b) in powder form ready for reconstitution with water, or (c) in a two-part or two-bottle format, where one bottle contains the hydrolysable tannins dissolved in propylene glycol or other suitable solvent, and the second bottle contains the water.
  • the two parts of this kit are then combined before first use.
  • the soothing, antioxidant and stabilizer agent(s) of the composition could be in either bottle, but it is simpler if they too are dissolved in the solvent. In that case, instead of providing a bottle containing just water, water can be added to the first bottle to make up the full volume, preferably in a pharmacy.
  • a liquid solution concentrate in glycerin, propylene glycol or any other suitable organic solvent or mixture of solvents.
  • This concentrated solution should contain tannic acid, Ectoine and excipients as mentioned above. Since the concentrated solution contains very low levels of water it is stable to hydrolysis. It would be diluted with water before use.
  • Inhalable fine powder formulations of the composition of the present invention can be prepared using one of more of the manufacturing steps known in the art and summarized in the article "Recent trends in DPI technology", Taneja Ruchi et al, International Research Journal of Pharmacy, 2012, 3 (11); hereby incorporated by reference.
  • the simplest powder formulation routes are either milling of the crystalline powders to the appropriate particle size, or spray drying - in both cases ensuring that a significant proportion of the powder particles fall in the respirable range of under 3.3 microns.
  • additional excipients optimized to enable absorption are added to the liquid mix before spray drying.
  • the thus prepared powders may be suitable for either pulmonary delivery to the lungs as aerosols, using a dry powder inhaler (DPI) or dry powder nebulizer. Additionally, there are also devices for dry-powder inhalation into the nasal cavity.
  • the composition of the present invention may be optimized for nasal or pulmonary inhalation, whether by use of a nasal spray, nebulizer or any other aerosol delivery device, and either as a liquid aerosol or a powder one. Additionally, use of this composition in nasal drops or as a throat spray is also possible.
  • the tannins serve as a natural anti-viral barrier providing wide-spectrum virucidal abilities; i.e. they constitute an agent (physical or chemical) that deactivates or destroys viruses.
  • the formulation of the present invention works in a different way from an antiviral drug, the latter serving to inhibit the development of the virus.
  • the mechanism of action of tannins in destroying viruses is thought to be the ability of these compounds to precipitate the proteins in the viral envelope provides the tannins with antiviral properties. In this way, tannins bond to viral (and also bacterial membranes) and destroy them.
  • compositions of the present invention represent a comfortable and effective means for the prophylaxis and/or treatment of influenza, rhinovirus, and RSV and/or of their symptoms. Additionally, these compositions can serve as a nasal aerosol for treating sinus infections.
  • the ability of the composition to decolonize bacterial infections in the lungs is important in two contexts: to prevent bacterial complications after a viral infection such as influenza, and to treat the microbial infections associated with cystic fibrosis (CF).
  • CF cystic fibrosis
  • common bacteria such as Staphylococcus aureus and Haemophilus influenzae colonize and infect the lungs.
  • Pseudomonas aeruginosa (and sometimes Burkholderia cepacia) dominates.
  • P. aeruginosa As tannins have been shown to be effective against both S. aureus and P.
  • aeruginosa it is expected that pulmonary delivery of the composition of the present invention will constitute an effective element of symptomatic relief for CF sufferers.
  • a mucolytic for example a natural mucolytic - such as N- Acetyl Cysteine and/or saline.

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Abstract

The present invention is in the field of virucidal tannin-comprising compositions for nasal and pulmonary delivery. Formulations of said compositions comprising soothing agents eleminating stinging effect of tannin are disclosed. Said compositions are intended for prohylaxis and / or treatment of influenza, viral rhinitis, cystic fibrosis and viral and bacteial lung infections.

Description

Virucidal Compositions
Background to the Invention
The broad spectrum antiviral nature of tannins is well known in the prior art (Mini Rev Med Chem. 2008 Oct; 8(12): 1179-87). Specifically, they are known from literature to be active against influenza. Tannins are virucidal compounds in the sense that they attack the membrane wall of the virus. Although the literature uses the terms antiviral and virucidal interchangeably, the more correct term for the activity of tannins is "virucidal" as the mode of action is one of destroying viruses on contact. While the virucidal properties of tannins in the form of tannic acid and tannin-containing solutions are well known, a 1953 paper was the first to explicitly describe the way in which influenza viruses are deactivated. This paper (Frisch A.W., Carson R.S., "Mode of inactivation of influenza virus by tannic acid", J Bacteriol. 1953 November; 66(5): 572-575.) describes how: "both the in vivo and in vitro inactivations of influenza virus are probably related to the potent protein denaturing action of tannic acid."
That this activity against influenza virus is a more general property of tannins was shown by Nakayama M., Suzuki K., Toda M., et al., "Inhibition of the infectivity of influenza virus by tea polyphenols" , Antiviral Res. 1993; 21; p.289-299. This paper shows how tannin products epigallocatechin gallate (EGCg) and theaflavin digallate (TF3) (1-10 μΜ) inhibited the infectivity of both influenza A virus and influenza B virus in Madin- Darby canine kidney (MDCK) cells in vitro. Their electron microscopy study showed how EGCg and TF3 (1 mM) agglutinated influenza viruses and prevented the viruses from adsorbing to MDCK cells. Their findings "suggest that tea polyphenols bind to the haemagglutinin of influenza virus, inhibit its adsorption to MDCK cells, and thus block its infectivity" .
Influenza hemagglutinin (HA) is a glycoprotein found on the surface of the influenza virus, and thus we see that the mode of deactivation of influenza viruses by tannins is the denaturing of the proteins on contact with the external membrane of the virus. In a recent in vivo trial, Liu and his co-workers (Liu S, Li H, Chen L, Yang L, Li L, Tao Y, et al., "Epigallocatechin-3- gallate inhibition of Epstein-Barr virus spontaneous lytic infection involves ERK1/2 and PI3-K/Akt signaling in EBV-positive cells", Carcinogenesis 2013; 34: 627-37) orally administrated BALB/c mice with EGCG at varied doses for 5 days. On the 3rd day of the treatment, the mice were infected with influenza A virus. EGCG at 40 mg/kg per day dramatically improved the survival rate of the experimental animals, and also decreased virus replication and the mitigation of viral pneumonia in the lungs.
Similarly, tannins are known to have antibacterial properties. An overview of both the antiviral and antimicrobial effects of tannins can be found in the article: "Antimicrobial and antiviral activity of hydrolysable tannins", Mini Rev Med Chem. 2008 Oct; 8(12): 1179-87, by Buzzini P. et al. The antimicrobial mechanism of tannins is thought to result from the astringent property of the tannin inducing complexation with enzymes or substrates on the membranes of the microorganisms (J. Antimicrob. Chemother. (2001) 48 (4): 487-491). In specific examples, the role of tannins in combatting S. Aureus and P. aeruginosa have been shown in H.Akiyama et al. "Antibacterial action of several tannins against S. aureus", JAC, 2001, 48, pp. 487-491 ; and S. Ghafourian et al, "Effect of tannin extract against Pseudomonas aeruginosa producing metallo beta-lactamas", J Biol Regul Homeost Agents. 2012 Apr-Jun; 26(2):231-6.
Unfortunately, tannins are knows to be astringent and, at effective concentrations, convey a stinging sensation on the tissue, where they are applied. Natural tannin is an acidic compound, and the pH of a concentrated aqueous solution of tannic acid is lower than 3.5. Aqueous acidic solutions cause a stinging sensation on mucosa and should be basified to pH>5 in order to be comfortable. Accordingly, the use of tannin preparations within the nasal cavity - for example as a nasal spray or nasal drops - is problematic. The other problem with liquid tannin formulations is their lack of stability due to hydrolysis. Thus, although the antiviral nature of tannins is known, there are no virucidal tannin nasal sprays using tannic acid in the market. On the contrary, in WO 2011/082835 Al it is stated that "hydrosoluble tannins are not suitable for topical applications" .
Accordingly, the objective of the present invention is to overcome the formulation problems associated with delivering tannins such that the practical implementation of tannin-based compositions in the form of aerosols for nasal and pulmonary delivery will be enabled.
Summary of the Invention
In one aspect, the present invention therefore comprises a comfortable, non-stinging aqueous formulation of tannins. This comfortable composition results from dissolving the tannins in a mostly aqueous solution together with one or more compatible soothing agents and, optionally, also one or more suitable stabilizing agents. The soothing agents that may be used for this purpose may be selected from the groups including osmolytes and demulcents (sometimes referred to as mucoprotective agents), but only where the specific soothing agent chosen does not react with tannic acid to form insoluble tannates. Preferably, said soothing agent will also be permitted for use in a nasal and/or pulmonary aerosol. Similarly, any stabilizing agent(s) used need to be compatible with tannic acid and allowed in a nasal and/or pulmonary aerosol.
As a soothing agent or demulcent serves to form a soothing, protective film or complex when administered onto a mucous membrane surface, it serves to counteract and thus allay the stinging or astringent effect of the chemical agent dissolved within it.
Advantageously, to the extent that the soothing agent also serves to preserve or provide stability to the tannins, said demulcent thereby solves both of the above-described formulation issues at the same time. In a still further aspect of the present invention, compositions are provided for the nasal administration of tannin-containing aerosols and for the pulmonary administration of tannin-containing aerosols, including inhalation solutions. Said compositions may be used for the purpose of combating viral infection in general, and influenza, rhinovirus, RSV and viral pneumonia specifically. Advantageously, as tannins are also known for their antibacterial characteristics, the use of the composition of the present invention will also cause decolonization of bacterial activity in the nasal cavity, lungs and sinuses. This is expected to reduce the risk of bacterial complications following an influenza or other viral infection and minimize the symptoms of diseases such as cystic fibrosis.
Brief description of the drawings
Figure 1 a shows the cytotoxicity of the pure 2% Ectoine solution.
Figure lb shows the cytotoxicity of the combination with 0.2% tannic acid.
Figure 2a shows the cytotoxicity of the pure 2% Ectoine solution on RLE cells.
Figure 2b shows the cytotoxicity of the combination with 0.2% tannic acid on RLE cells.
Figures 3a, 3b and 3c show the virucidal efficacy of the exemplary composition against influenza viruses A/CA/07/09 (H1N1), A/HK/1/68 (H3N2), and B/Lee/40 respectively.
Figure 4 shows a plot-graph of the cytotoxicity information from the nasal epithelial model on the same graph as data showing the virucidal efficacy against the three influenza viruses mentioned in Figs 3a-c.
Detailed Description of the Invention
Tannic acid as powder (Merck, USP/EP) was dissolved in water, at a number of concentrations in the 0.1% to 1% range. Due to the astringent or stinging effect caused by the spraying of this tannic acid solution into the nose, a number of potential soothing agents were examined for their potential addition to the composition.
Demulcents are a broad class of substances, including the synthetic demulcents methyl cellulose, glycerin or propylene glycol, the natural demulcents pectin and honey, and active extracts from Aloe Vera and other natural sources. Obviously, compounds that are available off-the-shelf in pharmaceutical grade (USP) are preferred choices for the composition of the present invention. Unfortunately, in preliminary work performed by the authors, the preferred demulcents glycerin and propylene glycol were both shown to be irritants when used in a nasal spray; whereas methyl cellulose made the formulation too viscous to spray.
Finding a compatible demulcent is challenging as tannic acid forms precipitates with a wide range of organic and inorganic compounds as di- and trivalent metal ions, amines, amino acids, peptides, proteins and some other polymers. Precipitating tannates remove the tannic acid from solution and thus reduces its bioavailability. For example, our experiments with tannic acid and seawater demonstrated precipitation of calcium and magnesium tannates from the solution, whereas at pH=6-7 the precipitation was more intensive than at pH=3-5. Thus, tannic acid is incompatible in solutions with many active ingredients and excipients. Looking then at the natural demulcents for a potentially compatible compound, Aloe Vera, which is available in USP grade (Aloway Natural Health Products, Haymeadow Crescent Faerie Glen, Pretoria East, South Africa) was tested for its applicability. This compound has proved to be an excellent soothing agent for nasal sprays in general; and indeed it is used for that purpose in the Otrimer Aloe Vera product from Novartis Consumer Health (Nyon, Switzerland) where it serves to prevent the stinging caused by a saline nasal spray. However, adding it to our tannic acid solutions caused a tannate precipitate to emerge from the solutions, and thus Aloe Vera was shown by experiment to be incompatible with tannic acid in solution.
The search for potential soothing agents was then extended to incorporate further classes of chemicals such as osmolytes, to evaluate whether they have potential as soothing agents within the context of a nasal spray. Osmolytes include sugar alcohols (myoinositol, mannitol, sorbitol), quaternary amines (such as taurine, choline, betaine, betaine- glycine) and Ectoines. Further candidates include glutamate, aspartate or proline, phosphatidylcholine, phosphatidylinositol diglycerol phosphate, phosphorylcholine, glycerophosphorylcholines, and amino acids such as glutamine, glycine and alanine. After experimentation with a range of osmolytes, unexpectedly and surprisingly it was found that adding Ectoine or Hydroxyectoine (medical grade, bitop AG, Witten, Germany), to the composition, serves to at least partially eliminate the stinging associated with the tannic acid solutions. Furthermore, the clarity of Ectoine-tannin solutions demonstrates that the Ectoine does not form an insoluble tannate as a result; this being a non-obvious and surprising experimental result.
As the virucidal effect of the tannic acid is dose-dependent, it was important to quantify the maximum concentration of tannic acid which - based on the above surprising experimental result - can be comfortably delivered as a nasal aerosol. To that end, a cell model using epithelial nasal cells was established and a range of combinations of tannic acid and Ectoine were tested on it for cytotoxicity. Cytotoxicity was evaluated by a 24 hour incubation of RPMI-2650 (human nasal cell line) with successive dilutions of the tannin containing nasal spray. The tannin nasal composition was diluted in cell culture medium, and there was also an untreated healthy control and an SDS treated "death control". Toxicity was determined via neutral red assay, where absorption was measured at 540 nm and a high absorption represents a healthy cell. As a control, an existing Ectoine nasal spray solution comprising just 2% Ectoine (labelled "SNS01") was also diluted in cell medium and measured as described above.
Figure la shows the in vitro cytotoxicity of a solution comprising just 2% Ectoine; this being a comfortable nasal spray product which is currently marketed for symptomatic relief of rhinitis. Figure lb, by way of comparison, shows the in vitro cytotoxicity of an exemplary composition of the present invention comprising 0.2% tannic acid + 2% Ectoine. Note that the combined composition is slightly more cytotoxic than the pure Ectoine one, but is still within reasonable limits. The comfort level of the above exemplary composition was then verified by testing on ten human subjects who each received a spray of this solution into both nostrils and did not experience any discomfort.
In order to evaluate the suitability of the composition of the present invention for pulmonary delivery, the same comparison was performed using RLE lung cells, where Figure 2a shows the cytotoxicity of the pure 2% Ectoine solution and Figure 2b shows the cytotoxicity of the combination with 0.2% tannic acid. This testing on lung epithelial cells showed a similar pattern to that seen with the nasal epithelial cells, and thus it can be concluded that (from an irritation perspective) the composition is going to be suitable for pulmonary administration as well.
This exemplary composition (0.2% tannic acid and 2% Ectoine) was then tested at successive dilutions in order to determine whether, over the tested concentration range, tannic acid performed as an effective virucidal compound. The cell model used was the CPE-based EC50 assay using MDCK cells, where the composition was tested against the three influenza viruses: A/CA/07/09 (H1N1), A/HK/1/68 (H3N2), and B/Lee/40. The assay comprised the following steps: MDCK cells were seeded in 96-well plates and incubated overnight. The next day serial dilutions of the test article (starting at 25% final concentration, 2-fold dilutions) as well as a control compound were prepared in medium. The growth medium was aspirated from the cells and the compound dilutions were added at 2x the final concentration for a one-hour incubation period. Afterwards the virus was added at an MOI of 0.01 and the cells were incubated for 6 days. The cells were then fixed and stained with crystal violet in glutaric dialdehyde solution. The optical density was determined and the EC50 was calculated using the uninfected (cells only) control as 0% CPE and the controls without compound (virus only) as 100% CPE using a 4-PL curve fit of the OD.
Figures 3a, 3b and 3c shows the virucidal efficacy of the exemplary composition against influenza viruses A/CA/07/09 (H1N1), A/HK/1/68 (H3N2), and B/Lee/40 respectively. Note that the composition exhibits efficient inhibition of all of these viruses at just a 25% concentration, and that the dose-response curves are similar to those of the standard antiviral drug Zanamivir (which is shown for reasons of comparison).
Figure 4 then plots the cytotoxicity information from the nasal epithelial model (above) on the same graph as data showing the virucidal efficacy against the three above influenza viruses (shown as a band between the two dashed lines). From this graph we a clear picture emerges of the therapeutic window of the composition of the present invention; at least for nasal delivery. The two solid vertical lines define the concentrations at which the composition will be tolerable but nevertheless effective. Looking at this area, it is reasonable to state that the concentration of tannic acid should be between 0.02% and 0.2%. Additionally, there may well also be some room for the patient to tolerate some slight discomfort in return for greater efficacy, and therefore the upper concentration range of tannic acid could be raised to 2% in such cases. Note also that immediately after application, the composition will immediately be diluted with biological liquid (mucus) and thus the concentration delivered would be reduced significantly by this dilution; causing an initial, higher tannic acid concentration to rapidly be diluted towards the 0.2% level.
As mentioned above, tannic acid is inherently unstable in aqueous solution. This instability results from both oxidation and hydrolysis of the tannic acid components. Accordingly, a number of approaches were used to address this stability problem: (1) reduction of the aqueous percentage of the composition by using a 75/25 water/propylene glycol mix instead of just water; (2) addition of one or more preservative agents, (3) addition of one or more antioxidants, (4) pH adjustment and (5) addition of one or more osmolytes; as detailed below:
1. The extent to which the composition of the present invention can be made "less aqueous" by using propylene glycol is, unfortunately, relatively limited as the propylene glycol itself was found to cause irritation of the nasal mucosa when used in concentrations above 25%.
2. A number of preservative agents are potential candidates to increase the stability of the composition of the present invention. According to a review of preservative agents currently used in nasal sprays, "/« vitro effects of preservatives in nasal sprays on human nasal epithelial cells" (Am J._Rhinol. 2008 Mar-Apr;22(2): 125-9. doi: 10.2500/ajr.2008.22.3154), of the two common preservatives, benzalkonium chloride and potassium sorbate, the former causes epithelial damage, and thus only the latter should be used. Our experiments also included propyl gallate, which is an antioxidant agent, often used as a preservative agent in foods, together with BHA and BHT. As tannic acid exists in equilibrium with gallic acid, without being bound by theory, it is expected that propyl gallate should demonstrate a high degree of compatibility with tannic acid and thus preserve it relatively well. Of the other candidates, such as methyl paraben, propyl paraben and povidone-iodine, the last is least preferred as it may iodize the tannic acid.
3. Antioxidant agents suitable for the composition of the present invention include ascorbic acid, tocapherol and glutathione.
4. At an optimal physiological pH level of pH6-pH7, the hydrolysis rate is much lower than at pH=3-4, and therefore the tannic acid solution which is typically in the pH3- pH4 range may be pH adjusted (for example by adding NaOH or NaC03) in order to reduce the hydrolysis rate and thus enhance the stability of the composition.
5. Some literature has shown the potential of osmolytes to reduce the rate of hydrolysis.
Examples of this literature include US patent application US20070092925 and "Effects of Osmolytes on the SLN1-YPD1-SSK1 Phosphorelay system from Saccharomyces cerevisiae" (Biochemistry. Aug 25, 2009; 48(33): 8044- 8050.doi: 10.1021/bi900886g). Thus it is expected that the inclusion of osmolytes in the composition of the present invention will enhance its stability against degradation due to hydrolysis.
By adopting this formulation approach, preferred exemplary compositions according to the present invention contain from 0.02% to 2% by weight of tannins (for example tannic acid) and include at least one soothing agent. In further preferred compositions, the concentration of tannin will be between 0.02% and 0.2%. In further embodiments, said compositions further comprise at least one antioxidant agent and at least one preservative agent. In some cases the antioxidant and the preservative may be the same agent. Similarly, the soothing agent may also serve as a preservative, in particular preserving against hydrolysis of the tannins.
Further to the experimentation described above, some preferred formulations of the composition of the present invention, are as follows:
1. 0.2% tannic acid, 2% Ectoine, 0.03% propyl gallate + 0.5% ascorbic acid
2. 0.2% tannic acid, 2% Ectoine, 0.03% propyl gallate + 2% ascorbic acid 3. 0.2% tannic acid, 2% Ectoine, 0.06% propyl gallate + 1% ascorbic acid
4. 0.2% tannic acid, 2% Ectoine, 0.09% propyl gallate + 1% ascorbic acid
5. 0.2% tannic acid, 2% Ectoine, 0.09% propyl gallate + 1% ascorbic acid
These above formulations may be made up in either a purely aqueous solution (i.e. 100% water) or, in a preferred embodiment, a water / propylene glycol mixture may be used. Advantageously, using the latter mixture was shown to reduce the rate of hydrolysis, thus enhancing the stability of the product. The water / propylene glycol ratios used for such a mixture can range from 99%/l% to 75%/25%; as using above 25% propylene glycol makes the composition too viscous to spray well and somewhat painful to the nasal tissues. Accordingly, some preferred formulations incorporating propylene glycol are those of composition no. 2 above, in a 75/25 mixture or an 80/20 mixture.
As will be obvious to one skilled in the art, further suitable antioxidant agents and stabilizers can be added to the above compositions. Additionally, for a nasal composition, oil of mint, menthol, eucalyptus oil and/or saline or sea water may be added to the composition, to open the nasal passages.
The term "aerosol" as used herein refers to a colloid of fine solid particles or liquid droplets, in air or another gas. The composition of the present invention may be formulated either for use as a liquid-droplet aerosol or as a fine -powder aerosol.
Liquid droplets may, for example, be delivered using a spray bottle for nasal administration, or by a nebulizer for pulmonary administration. In both of these cases, the liquid formulation of this composition can be provided to the patient in a number of forms: (a) in final liquid form, ready for aerolization; (b) in powder form ready for reconstitution with water, or (c) in a two-part or two-bottle format, where one bottle contains the hydrolysable tannins dissolved in propylene glycol or other suitable solvent, and the second bottle contains the water. The two parts of this kit are then combined before first use. The soothing, antioxidant and stabilizer agent(s) of the composition could be in either bottle, but it is simpler if they too are dissolved in the solvent. In that case, instead of providing a bottle containing just water, water can be added to the first bottle to make up the full volume, preferably in a pharmacy.
Hydrolysis of tannic acid in aqueous solution during prolonged storage as per (b) above can be avoided using anhydrous mixture of tannic acid and Ectoine. Such mixtures are stable enough to be stored long period of time and could be reconstituted with water before use. Two types of anhydrous mixtures are suggested;
1. A solid mixture of tannic acid, Ectoine and excipients (optional) as preservatives, pH and osmotic modulators. This mixture could be prepared by dry blending, lyophilization, spray drying or co-milling of ingredients.
2. A liquid solution concentrate, in glycerin, propylene glycol or any other suitable organic solvent or mixture of solvents. This concentrated solution should contain tannic acid, Ectoine and excipients as mentioned above. Since the concentrated solution contains very low levels of water it is stable to hydrolysis. It would be diluted with water before use.
Inhalable fine powder formulations of the composition of the present invention can be prepared using one of more of the manufacturing steps known in the art and summarized in the article "Recent trends in DPI technology", Taneja Ruchi et al, International Research Journal of Pharmacy, 2012, 3 (11); hereby incorporated by reference. Broadly speaking, the simplest powder formulation routes are either milling of the crystalline powders to the appropriate particle size, or spray drying - in both cases ensuring that a significant proportion of the powder particles fall in the respirable range of under 3.3 microns. In some cases, additional excipients optimized to enable absorption are added to the liquid mix before spray drying. The thus prepared powders may be suitable for either pulmonary delivery to the lungs as aerosols, using a dry powder inhaler (DPI) or dry powder nebulizer. Additionally, there are also devices for dry-powder inhalation into the nasal cavity.
Thus, the composition of the present invention may be optimized for nasal or pulmonary inhalation, whether by use of a nasal spray, nebulizer or any other aerosol delivery device, and either as a liquid aerosol or a powder one. Additionally, use of this composition in nasal drops or as a throat spray is also possible. In all of these cases, without being bound by theory, the tannins serve as a natural anti-viral barrier providing wide-spectrum virucidal abilities; i.e. they constitute an agent (physical or chemical) that deactivates or destroys viruses. In this way, the formulation of the present invention works in a different way from an antiviral drug, the latter serving to inhibit the development of the virus. The mechanism of action of tannins in destroying viruses is thought to be the ability of these compounds to precipitate the proteins in the viral envelope provides the tannins with antiviral properties. In this way, tannins bond to viral (and also bacterial membranes) and destroy them.
As described above, the compositions of the present invention represent a comfortable and effective means for the prophylaxis and/or treatment of influenza, rhinovirus, and RSV and/or of their symptoms. Additionally, these compositions can serve as a nasal aerosol for treating sinus infections.
Furthermore, the ability of the composition to decolonize bacterial infections in the lungs is important in two contexts: to prevent bacterial complications after a viral infection such as influenza, and to treat the microbial infections associated with cystic fibrosis (CF). In the initial stage of CF, common bacteria such as Staphylococcus aureus and Haemophilus influenzae colonize and infect the lungs. Eventually, Pseudomonas aeruginosa (and sometimes Burkholderia cepacia) dominates. By 18 years of age, 80% of patients with classic CF harbor P. aeruginosa. As tannins have been shown to be effective against both S. aureus and P. aeruginosa (see above), it is expected that pulmonary delivery of the composition of the present invention will constitute an effective element of symptomatic relief for CF sufferers. In order to reduce the levels of mucus in the lungs, it may be useful to also add a mucolytic to the composition, for example a natural mucolytic - such as N- Acetyl Cysteine and/or saline.

Claims

Claims:
1. A virucidal composition for nasal and/or pulmonary administration, said composition comprising tannins together with at least one soothing agent; such that said soothing agent at least partially eliminates the stinging due to the tannins.
2. The composition of claim 1, wherein said composition is formulated as a mostly aqueous liquid for delivery as an aerosol via a nasal spray and/or a nebulizer.
3. The composition of claim 1, wherein said soothing agent is selected from the group comprising of osmolytes, compatible solute, demulcents and any combination thereof.
4. The composition of claim 3, wherein said osmolytes include Ectoine and/or Hydroxyectoine .
5. The composition of claim 1, wherein said composition further comprises a solvent such as prolylene glycol to reduce hydrolysis of the tannins.
6. The composition of claim 1 , wherein said tannins include tannic acid.
7. The composition of claim 1, further comprising antioxidant agents, including ascorbic acid, tocapherol, and glutathione.
8. The composition of claim 1, further comprising preservative agents, including potassium sorbate, propyl gallate, methyl paraben, propyl para ben and povidone- iodine.
9. The composition of claim 7, wherein said antioxidant agents also comprise preservative characteristics.
10. The composition of claim 1, wherein said soothing agent also comprise preservative characteristics.
11. The composition of claim 1, where said tannins comprise between 0.02% and 2% by weight of said composition.
12. The composition of claim 1, where said tannins comprise between 0.02% and 0.2% by weight of said composition.
13. The composition of claim 1, further comprising one or more selected from the group consisting of oil of mint, menthol, eucalyptus oil, saline or sea water and any combination thereof; for use as a nasal spray.
14. The composition of claim 1, for prophylaxis and/or treatment of viral conditions including influenza, rhinitis, RSV and viral pneumonia.
15. The composition of claim 1, for prophylaxis and/or treatment of sinusitis.
16. The composition of claims 1, further comprising a mucolytic, such as natural mucolytic N-Acetyl Cysteine and/or saline; for treating lung conditions where the lungs are colonized by bacteria; for example cystic fibrosis.
17. The composition of claim 1, wherein said composition is provided as a dry powder product which is reconstituted with water.
18. The composition of claim 17, provided in a two-part water / solvent format, using a solvent such as propylene glycol, where the tannins are dissolved in the solvent part; thereby enabling an extended shelf life.
19. The composition of claim 1, where said composition is formulated as an inhalable dry powder aerosol, for delivery via a dry powder inhaler or dry powder nebulizer.
20. A method for manufacturing a virucidal composition for nasal and/or pulmonary administration, comprising steps of:
a) adding of tannins from about 0.02% to about 3% by weight to water or to a water- propylene gycol mixture or a water-glycerin mixture in a volume ratio of about 99: 1 to about 50:50; and stirring to dissolution;
b) adding of a soothing agent to the solution prepared from about 0.01% to about 3% and stirring to dissolution;
c) adding of preservatives to the solution prepared, from about 0.01% to about 1%, and / or ascorbic acid from about 0.1% to about 3%;
d) adjustment of the pH to the range 5-7;
such that said soothing agent at least partially eliminates the stinging due to the tannins.
21. The method of claim 20 wherein said soothing agent is Ectoine.
22. The method of claim 20 wherein said preservatives include propyl gallate
23. The method of claim 20, wherein said method produces a composition which is formulated as a mostly aqueous liquid for delivery as an aerosol via a nasal spray and/or a nebulizer.
24. The method of claim 20, wherein said soothing agent is selected from the group comprising of osmolytes, compatible solute, demulcents and any combination thereof.
25. The method of claim 22, wherein said osmolytes include Ectoine and/or Hydroxyectoine .
26. The method of claim 20, further comprising a step of adding a solvent such as prolylene glycol to reduce hydrolysis of the tannins.
27. The method of claim 20, wherein said tannins include tannic acid.
28. The method of claim 20, further comprising a step of adding antioxidant agents, including ascorbic acid, tocapherol, and glutathione.
29. The method of claim 20, further comprising a step of adding preservative agents, including potassium sorbate, propyl gallate, methyl paraben, propyl paraben and povidone-iodine.
30. The method of claim 27, wherein said antioxidant agent also comprises preservative characteristics.
31. The method of claim 20, wherein said soothing agent also comprises preservative characteristics.
32. The method of claim 20, further comprising a step of adding one or more of the group consisting of oil of mint, menthol, eucalyptus oil, saline or sea water and any combination thereof; for use as a nasal spray.
33. The method of claim 20, wherein said composition is used for prophylaxis and/or treatment of viral conditions including influenza, rhinitis, RSV and viral pneumonia.
34. The method of claim 20, wherein said composition is used for prophylaxis and/or treatment of sinusitis.
35. The method of claim 20, further comprising a step of adding a mucolytic, such as natural mucolytic N-Acetyl Cysteine and/or saline; for treating lung conditions where the lungs are colonized by bacteria; for example cystic fibrosis.
36. The method of claim 20, wherein said method further comprises a step of lyophilizing said composition to a dry powder product which is reconstituted with water.
37. The method of claim 34, further comprising a step of reconstituting said dry powder in a two-part water / solvent format, using a solvent such as propylene glycol, where the tannins are dissolved in the solvent part; thereby enabling an extended shelf life.
38. The method of claim 20, further comprising a step of formulating said composition as an inhalable dry powder aerosol, for delivery via a dry powder inhaler or dry powder nebulizer.
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