FIELD

This disclosure relates to compositions comprising nitric oxide and uses, methods, and devices thereof. BACKGROUND

Nitric oxide (NO) is produced in the endothelium tissue of the human body as part of normal physiological processes. NO is an endogenous vasodilator i.e., an agent that widens the internal diameter of blood vessels. NO has also been investigated for its use as a sterilizing agent. It has been discovered that NO will interfere with or kill the growth of bacteria grown irt vitro. WOOO/30659 discloses a method and apparatus for the treatment of respiratory infections by NO inhalation. It has also been suggested that NO has an inhibitory effect on the life cycle of the influenza virus. See, for example, Rimmelzwaan et. al, Journal of Virology; Vol. 73, No. 10; p. 8880-8883 (Oct. 1999) and Akerstrom et. al, Journal of Virology; Vol. 79, No. 3; p. 1966-1969 (Feb. 2005). In addition, it has been suggested that NO gas may be used to treat various conditions (US7,520,866; US6,432,077; US6,793,644; US7,122,018; EP1,283,724; Tucker A. T. et. al. Lancet (1999) Nov 13;354:1670-5; J. B. J. Hardwick et. al, Clinical Science (2001) 100, 395^100).

SUMMARY

The present disclosure provides gel compositions comprising nitric oxide. It has been found that NO can be formulated into a gel composition and maintain its anti-microbial effect.

The present disclosure also provides uses, methods, and devices that utilize NO- containing gels.

The present disclosure also provides compositions comprising greater than 1 mM to less than 100 mM of nitrite.

The compositions of the present disclosure have an anti-microbial and/or sterilizing activity. It has surprisingly been found that, for optimal effect, the concentration and/or pH of the compositions should be kept within relatively narrow ranges.

As used herein 'gel' refers to compositions having a continuous liquid phase and comprising a cross-linked network. The compositions herein preferably have a viscosity at viscosity at 21°C of from about 10 mPa-s to about 50000 mPa-s; about 20 mPa-s to about 40000 mPa-s; 30 mPa-s to about 30000 mPa-s; 40 mPa-s to about 20000 mPa-s; 50 mPa-s to about 15000 mPa-s.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows the effect of 5mM and lOmM nitrite solutions on Trichophyton Rubrum (1), Trichophyton Mentagrophytes (2), Candida albicans (3), and Aspergillus Niger (4). The 5mM solution significantly reduces the number of plaques for Trichophyton Rubrum (1), Trichophyton Mentagrophytes (2), and Aspergillus Niger (4). The lOmM solution leaves no plaques for Trichophyton Rubrum (1), Trichophyton Mentagrophytes (2), and Aspergillus Niger (4). In addition, the number of plaques are reduced for Candida albicans (3).

Figure 2 shows the effect of 1 mM and 10 mM nitrite sprays of Influenza A (Common Flu). Column A was treated with two sprays of 1 mM nitrite solution and Column B was treated with two sprays of 10 mM nitrite solution.

Figure 3 shows the decay in the [N02] (mM) of 0.3 gel viscosity over a 6 day period is slower relative to the change in nitirite concentration with time for the liquid phase.

Figure 4 shows the decay in the nitrite concentration over time in an enclosed vial.

DETAILED DESCRIPTION

The present compositions comprise nitric oxide. The present compositions may be prepared from solvents which have been exposed to a suitable amount of nitric oxide has been dissolved in a suitable solvent. For example, saline treated with NO has been found to inhibit microbial transmissibility. Therefore, it is within the scope of this disclosure to prepare a composition from NO in a solvent. Any suitable solvent or mixture of solvents may be used herein. For example, NO is soluble in water and various alcohols such as methanol, ethanol, isopropanol, and the like. In an embodiment the solvent is water, for example, in the form of saline. One aspect of the present disclosure relates to an anti-viral composition comprising a nitric oxide solution as described herein.

While not wishing to be bound by theory, it is believed that the NO forms nitrite in solution which, upon application, devolves back into NO gas which has an antimicrobial effect.

2 The present gel compositions may, for example, be prepared by adding a gelling agent to a NO-containing liquid or exposing existing gels to NO at an appropriate concentration for an appropriate amount of time. For example, the gel composition may be exposed to a concentration of from 100 ppm to 20,000 ppm for approximately 30 seconds.

While not wishing to be bound by theory, it is believed that the present gel compositions will release NO over a period of time providing a 'slow-release' mechanism which delays or controls the anti-microbial/anti-parasitic effect. Moreover, when for example used topically it is believed that the present gels will provide a longer-lasting effect because they persist on the skin for a longer period. This can, for example, be beneficial in combating certain parasitic conditions such as leishmaniasis.

In an embodiment of the present disclosure there is provided an NO composition having an appreciable anti-microbial/anti-parasitic effect for 2 hours or more; 4 hours or more; 6 hours or more.

The present disclosure provides compositions comprising NO in a therapeutically acceptable amount i.e. an amount that is sufficient to achieve the therapeutic purpose for which the composition is intended. For example, the concentration of NO in solution is preferably from about 10 μΜ or greater, or from about 50 μΜ or greater, or from about 100 μΜ or greater. In an embodiment the concentration of NO in solution is from about 100 raM or less, or from about 75 mM or less, or from about 60 mM or less, or from about 25 mM or less.

The present disclosure provides compositions comprising greater than about 1 mM and less than about 100 mM of NO. Preferably the concentration is from about 1.5 mM to about 80 mM. More preferably the concentration is from about 2 mM to about 70 mM. Even more preferably the concentration is from about 3 mM to about 25 mM. Even more preferably the concentration is from about 4 mM to about 10 mM.

As can be seen from Figure 4, even within a closed system the nitrite concentration decreases over time until it reaches a relatively steady state. Unless otherwise specified, as used herein references to NO concentration refers to the concentration prior to use of the composition. It can be seen that it may be necessary to introduce a higher concentration of NO during the formulation of the gel in order to provide a gel composition having the appropriate concentration of NO at the time of usage. The present disclosure provides compositions having a pH of from about 3 to about 6, or from about 3.2 to about 5.5, or from about 3.4 to about 4.5, or from about 3.5 to about 4.

The compositions herein may be prepared from exposing gels or solutions to NO gas or substances that can devolve nitric oxide. For example, the composition may comprise nitrites. Common nitrite salts that may be useful in the present composition include sodium nitrite, potassium nitrite, calcium nitrite, other alkali or alkali earth nitrites, or suitable combinations thereof. Sodium nitrite may be used herein.

The present gels may be prepared in any suitable manner. For example, the compositions may comprise gelling agents. Numerous gelling agents are known in the art. Examples include, but are not limited to, polyvinylpyrrolidone, copolymers of a vinylpyrrolidone, polyethylene oxide, methacrylamide, N-vinylimidazole, polyvinyl alcohol, (meth)acrylate polymers, agarose, hydroxymethylcelluloses, hydroxyethylcelluloses, hyaluronan, gelatin, glycerine, silicic acid, natural gums, agar, starches, pectins, alginates, alginic acid, carrageenan, Locust bean gum, and suitable combinations thereof. For example, the gelling agent may be selected from hydroxymethylcelluloses, hydroxyethylcelluloses, glycerine, gun arabic, alginate, or combinations thereof. The present gels preferably comprising from about 0.1 % to about 20%, by weight, of gelling agent. More preferably the present gels comprise from about 0.5% to about 10%, by weight, of gelling agent. In an example, 1% of hydroxyethylcellulose is used. In another example, 1% of hydroxymethylcellulose is used. In another example, 5% of glycerine is used

The present disclosure provides a hydrogel composition comprising nitric oxide and having a viscosity at 21°C of from about 5 mPa-s to about 100000 mPa-s. For example, the gel compositions herein may have a viscosity at 21°C of from about 10 mPa-s to about 50000 mPa s; about 20 mPa s to about 40000 mPa s; 30 mPa-s to about 30000 mPa-s; 40 mPa-s to about 20000 mPa-s; 50 mPa-s to about 15000 mPa-s.

The present disclosure provides for modifying currently marketed gel products by the addition of nitric oxide. For example, nitric oxide or substances that can devolve nitric oxide can be added to water-soluble lubricants like KY Jelly® or to petroleum jellies like Vaseline®. The present disclosure provides a spray composition comprising NO. For example, the present disclosure provides a nasal spray which may be used to combat respiratory infections such as colds and flu. The spray may have a lower viscosity than that of the above mentioned gels. The present spray preferably has a pH of from about 3 to about 6, or from about 3.2 to about 5.5, or from about 3.4 to about 4.5, or from about 3.5 to about 4. The present spray preferably has a concentration of NO in solution is from about 10 μΜ or greater, or from about 50 μΜ or greater, or from about 100 μΜ or greater. In an embodiment the concentration of nitrites/nitrates in solution is from 10 mM or less, or from about 1 mM or less, or from about 500 μΜ or less.

The present disclosure provides a wash composition comprising NO. For example, the wash may be used to combat diabetic leg ulcers, fungal infections, ear infections, sore thoat (due to Streptococci bacteria or viruses)or other persistent skin infections. The wash may also be used for sterilization, antiseptic, or prophylactic purposes. The spray may have a lower viscosity than that of the above mentioned gels. The present spray preferably has a pH of from about 3 to about 6, or from about 3.2 to about 5.5, or from about 3.4 to about 4.5, or from about 3.5 to about 4.

The NO concentration in the present wash is preferably greater than about 1 mM and less than about 100 mM of NO. Preferably the concentration is from about 1.5 mM to about 80 mM. More preferably the concentration is from about 2 mM to about 70 mM. Even more preferably the concentration is from about 3 mM to about 25 mM. Even more preferably the concentration is from about 4 mM to about 10 mM.

It has been found that certain the present compositions are relatively stable and may be stored for a period before use. Further, it has been found that certain of the present compositions may provide a sustained-release of the anti-microbial NO over a period of time.

The present compositions may comprise various other ingredients. Examples include, but are not limited to, acne preparations, analgesics, anaesthetics, antibiotics, antihistamines, anti-infective agents, anti-inflammatory agents (e.g corticosteroids, NSAIDs), antipruritics, anti-psoriasis agents, antipsychotics, antipyretics, anti-seborrhoeic agents, antiviral agents, burn preparations, cardiovascular agents (e.g. e.g., alpha or beta blockers), cleansing agents, deodorants, depigmenting agents, diaper-rash products, emollients, hair-growth products, hormones (e.g. androgens, estrogens, progesterone), keratolytics, lubricants, mast cell stabilizers, moisturizers, mouth wash, pediculicides, photosensitizing agents, scabicides, steroids, sunburn preparations, sunscreens, surfactants, vasoconstrictors, vasodilators, vitamins, wet dressings, wound care products, and combinations thereof. The present compositions may comprise eucalyptus oil. The present compositions may comprise a variety of optional ingredients. The CTFA Cosmetic Ingredient Handbook, Second Edition (1992) describes a wide variety of non- limiting, widely-used cosmetic and pharmaceutical ingredients, which are suitable for use in the present compositions. For example, preservatives, emollients, solvents, humectants, powders, kaolin, starch, gums, silicas, oils, emulsifiers, water, surfactants, preservatives, buffers, waxes, plant extracts, thickeners, activity enhancers; colourants, perfumes, sunscreens, fatty acids, polyols, hydroxy carboxylic acids, keto carboxylic acids, sterols, chelating agents, thickening agents, colourants, and combinations thereof.

The present compositions may have non-Newtonian or shear-thinning properties such that an appropriate viscosity is maintained while the composition is stored in a container and then upon dispensing from the container, the viscosity becomes lower to facilitate dispensing and spreading of the composition on skin or hair of the consumer. The present compositions may be packaged in a variety of containers. Preferred containers include pressurized containers, pump containers, tubes, sachets, and the like. In a preferred embodiment the present gel compositions are stored in a pressurized container prior to use.

The present compositions may be used to treat any suitable condition. For example, the present compositions may be used to treat chronic non-healing wounds, abscesses, ulcers, acute wounds, MRSA infections, fungal infections, bacterial infections, viral infections, parasitic infections, and the like.

Other conditions which may benefit from treatment with the present compositions include, but are not limited to skin conditions such as those caused by Staphylococcus spp. (e.g. Folliculitis, Furunculosis, Impetigo, Staphylococcal scalded-skin syndrome, Toxic Shock syndrome); Streptococcus spp. (e.g. Cellulitis, Erysipelas, Impetigo, Necrotising fasciitis, Scarlet fever); Corynebacterium spp. (e.g. erythrasma, pitted keratolysis, trichomycosis axillaris); Gonorrhoea; Meningococcal disease; Erysipeloid; Chancroid; Urticaria; Rosacea; Rhionscleroma; Athlete's foot; Gram-negative folliculitis; Chronic paronychia; Spa-pool folliculitis; Eecthyma gangrenosum; Anthrax; Granuloma inguinale; Gangrene; Syphilis; Yaws; Pinta; Lyme's disease; Bacillary angiomatosis; Bartonellosis; Leprosy; Bacterial vaginosis; Leishmaniasis; Kawasaki disease; Pseudofolliculitis barbae; Sarcoidosis; Bacterial endocarditis; Morbilli; Rubella; Varicella; Fifth Disease; Roseola; Mononucleosis; Glandular fever; Enterovirus infections; Pityriasis rosea; Hand foot and mouth disease; Gianotti-Crosti syndrome; Laterothoracic exanthema; Haemorrhagic fevers; Smallpox; Cowpox; Chikungunya fever; Dengue fever; viral warts (e.g verrucas, genital warts, condylomas, squamous cell papillomas); Influenza; Leishmaniasis; Malaria; Skin cancers or other types of cancer; Kaposi sarcoma; Rickettsial diseases; Yellow fever; HIV; Herpes simplex; Herpes zoster; Vesicular stomatitis; Molluscum contagiosum; Orf; or Milker's nodules.

The present compositions may be used to treat infections that are unresponsive to antibiotic treatment.

The present compositions may be used to prophylatically treat a subject at risk of developing an infection. For example, the present compositions may be applied to a subject who has been or will be exposed to an infectious agents such as bacteria, virus, fungi, or parasites.

In an embodiment the present compositions may be used as a prophylactic to reduce the risk of a sexually transmitted disease such as HIV or gonorrhoea from being transmitted.

The present compositions may also be applied to surfaces in order to sterilize said surfaces or to render them inhospitable to infectious agents.

The present disclosure provides a method of treatment of a human or animal subject.

The compositions may be applied directly to the area to be treated or may be applied in conjunction with a dressing. For example, in an embodiment of the present method a composition according to the present disclosure is applied directly to a non-healing wound on the skin of a mammal. The wound is then covered with a dressing. The present compositions may also be applied as a hand wipe, or applied into the nares of the nose, as a vaginal or penile lubricant. The present compositions may be applied to the udders of cows to treat mastitis or their nares to treat bovine viral respiratory complex.

The compositions herein may be produced by infusing NO into saline and then adding an appropriate gelling agent. The compositions herein may be produced by adding an appropriate gelling agent to a solution and then infusing NO into the resultant mixture. For example, the gelling agent may be glycerine, hydroxmethylcellulose, hydroxethylcellulose, or combinations thereof.

EXAMPLES

Four different fungi, Aspergillus Niger (32656), Candida albicans (14053), Trichophyton Mentagrophytes (114841) and Trichophyton Rubrum (18758), were plated on Sabouraud Dextrose Broth + Agar plates and grown to spore stage. Glass beads were put on the plates and then shaken for 5 minutes. The beads from each plate were then transferred into four 25 ml vials containing 5 ml of Sabouraud Dextrose Broth and grown at 30°C for 24- 48 hours. The media was then diluted to 25 ml diluted with media - Sabouraud Dextrose Broth to final volume of 25 ml. ΙΟΟμί of the fungi-infected media was then transferred into 3 mL saline containing either 5 or 10 mM nitrite. The pH of the solutions was 3.7. The inoculates were then incubated for 30 minutes at 30°C. ΙΟΟμί of this solution was then plated Sabouraud Dextrose Broth + Agar plates and incubated for 72 hours.

The 5 mM solutions significantly reduced the amount of viable fungi for Aspergillus Niger (32656), Trichophyton Mentagrophytes (114841) and Trichophyton Rubrum (18758). The lOmM solutions eradicated Aspergillus Niger (32656), Trichophyton Mentagrophytes (114841) and Trichophyton Rubrum (18758), and markedly reduced Candida albicans (14053).