Synergistic antimicrobial preparations containing chlorite and hydrogen peroxide

Abstract

An anti-microbial preservative for use in ophthalmic and dermatologic products. The preservative includes from about 0.005 wt. % to about 0.20 wt. % chlorite compound and from about 0.005 wt. % to about 0.05 wt. % peroxy compound. Additionally, the preservative does not generate chlorine dioxide within the pH range of 5.0-8.8. Also included are an antimicrobial ophthalmic and dermatologic compositions for direct application onto an eye or skin of a living being including from about 0.005 wt. % to about 0.20 wt. % chlorite compound and from about 0.005 wt. % to about 0.05 wt. % peroxy compound. Also included are methods for treating dryness of the eyes and skin disorders (e.g., wounds, burns, infections, ulcerations, psoriasis, etc.) and for disinfecting and cleansing contact lenses while in place upon an eye by applying the composition to the eye or to the contact lens.

Claims

1. An anti-microbial ophthalmic composition, the composition consisting from about 0.001 wt. % to about 0.20 wt. % chlorite compound and from about 0.0002 wt. % to about 0.05 wt. % peroxy compound, wherein the composition does not contain chlorine dioxide prior to contact with the eye of an individual, and wherein the ophthalmic composition is at a pH range of between 5.0 and 8.8.

2. The anti-microbial ophthalmic composition of claim 1, wherein the chlorite compound is a metal chlorite.

3. The anti-microbial ophthalmic composition of claim 2, wherein the metal is sodium.

4. The anti-microbial ophthalmic composition of claim 2, wherein the metal is metal is selected from the group consisting of potassium, calcium, and magnesium.

5. The anti-microbial ophthalmic composition of claim 1, wherein the peroxy compound is hydrogen peroxide.

6. The anti-microbial ophthalmic composition of claim 1, wherein the composition is applied onto an eye for treating dryness of an eye.

7. The anti-microbial ophthalmic composition of claim 1, wherein the composition is applied onto an eye for treating an infection of an eye.

8. The anti-microbial ophthalmic composition of claim 7, wherein the infection is caused by a bacterial keratitis.

9. The anti-microbial ophthalmic composition of claim 7, wherein the infection is caused by a virus.

10. The anti-microbial ophthalmic composition of claim 7, wherein the infection is caused by a fungus.

11. The anti-microbial ophthalmic composition of claim 1, wherein the composition is applied onto a contact lens in place on the eye.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a graph demonstrating the non-production of chlorine dioxide at room temperature in the chlorite/peroxide preparation of the present invention at pH level 7.3;

(2) FIG. 2 is a graph demonstrating the non-production of chlorine dioxide at room temperature in the chlorite/peroxide preparation of the present invention at pH level 8.0;

(3) FIG. 3 is a graph demonstrating the non-production of chlorine dioxide at room temperature in the chlorite/peroxide preparation of the present invention at pH level 8.8;

(4) FIG. 4 is a graph demonstrating the non-production of chlorine dioxide at room temperature in the chlorite/peroxide preparation of the present invention at pH level 7.0;

(5) FIG. 5 is a graph demonstrating the non-production of chlorine dioxide at room temperature in the chlorite/peroxide preparation of the present invention at pH level 6.44;

(6) FIG. 6 is a graph demonstrating the non-production of chlorine dioxide at room temperature in the chlorite/peroxide preparation of the present invention at pH level 6.0; and

(7) FIG. 7 is a graph demonstrating the production of chlorine dioxide at room temperature in the chlorite/peroxide preparation of the present invention at a pH level of 1.5.

DETAILED DESCRIPTION

(8) The following detailed description and examples are provided for the purpose of describing certain exemplary embodiments of the invention only, and are not intended to limit the scope of the invention in any way.

(9) The present invention provides preparations which contain chlorite (e.g., a metal chlorite such as sodium chlorite) in combination with a small amount of hydrogen peroxide in neutral aqueous (pH 5.0-8.8, preferably pH 7.0-7.8, and more preferably pH 7.0-7.4) solution. These preparations exhibit synergistic antimicrobial activity without generating chlorine dioxide during storage at room temperature, thereby rendering the stability of these solutions acceptable for pharmaceutical use. For example, an aqueous solution containing 400 ppm chlorite plus 100 ppm hydrogen peroxide remains stable beyond 18 months at room temperature, and is effective to reduce Candida albicans activity by 1.0 log within six hours of challenge, even though the individual components of such solution are ineffective when applied separately at the same concentrations to reduce Candida albicans activity. Additionally, the hydrogen peroxide present within the chlorite/peroxide solutions of the present invention readily decomposes into molecular oxygen and water, upon contact with the peroxidase and catalase enzymes present in tissue and/or some body fluids. Such in situ generation of molecular oxygen contributes to cell vitality and enhances wound healing.

(10) The chlorite/H.sub.2O.sub.2 solutions of the present invention are sufficiently stable to be formulated in combination with polymeric lubricants (non-ionic and/or anionic; e.g., HPMC, Methocel, CMC, hyaluronic acid, etc.,) and/or in combination with block polymer based surfactants (e.g., pluronics). For example, an aqueous chlorite/hydrogen peroxide system can be formulated together with methocel or hyaluronic acid as a lubricant and pluronics as a surfactant for contact lens disinfectant solution (viscosity up to 50 cps at 25 degrees C.) in an ophthalmically acceptable tonicity (e.g., osmolality of at least about 200 mOsmol/kg) and a buffer to maintain the pH of the formulation within an acceptable physiological range. The formulation of the contact lens disinfection solution, artificial tear solution, and in-eye cleaner solution, contains chlorite preferably from about 0.005 to about 0.06 weight/volume percent and hydrogen peroxide preferably from about 0.0002 to about 0.05 weight/volume percent. Again, the presence of hydrogen peroxide provides the beneficial oxygen molecule to the cornea upon contact with catalase in the tear.

A. Formulations

(11) The chlorite/peroxide preparations of the present invention may be formulated in various ways, including liquid solutions, gels, ointments, creams, sprays, etc. Set forth herebelow are a few examples of the types of specific formulations which may be prepared in accordance with this invention.

(12) i. Stable Chlorite/Peroxide Liquid Solutions

(13) The following Formula 1 is a first preferred formulation of a liquid chlorite/peroxide solution of the present invention:

(14) TABLE-US-00001 FORMULA 1 Sodium Chlorite 0.005%-0.10% Hydrogen Peroxide 0.005%-0.05% Methocel A 0.05%-0.2% Boric Acid 0.15% Sodium Chloride 0.75% Pluronic F-68/F-127 0.1% HCl or NaOH Adjust pH 7.4 Purified water Q.S. to volume

(15) The following Formula 2 is a second preferred formulation of a liquid chlorite/peroxide solution of the present invention:

(16) TABLE-US-00002 FORMULA 2 Sodium Chlorite 0.05% Hydrogen Peroxide 0.02% Carboxymethyl Cellulose 0.01% Boric Acid 0.15% Sodium Chloride 0.75% Pluronic F-68/F-127 0.1% HCl or NaOH Adjust pH 7.3 Purified water Q.S. to volume

(17) The chlorite/peroxide solutions of the present invention, such as the solution of the above-shown preferred formulation, may be used for a variety of medical and non-medical applications including but not necessarily limited to a) disinfection of articles and surfaces such as contact lenses, medical/dental instruments, counter tops, treatment tables, combs and brushes, etc.; antisepsis of skin or body parts (e.g., a disinfectant hand wash, antiseptic facial scrub, etc.,) and b) treatment or prophylaxis of dermal (i.e., skin or mucous membrane) disorders such as wounds, burns, infections, ulcerations, cold sores, psoriasis, acne, and c) deterrence or prevention of scar formation, and d) treatment of ophthalmic disorders (e.g., infections or inflammations caused by bacterial keratitis).

(18) As pointed out earlier, the chlorite/hydrogen peroxide system of the present invention is sufficiently stable to be formulated in a polymeric gel form or in a paste form. Furthermore, such polymeric gel or paste formulation can contain polymers which delay or control the release of the chlorite/hydrogen peroxide (e.g., a sustained release delivery system). Such sustained release formulations provide outstanding benefits of increasing therapeutic index by maintaining the effective concentration of chlorite/H.sub.2O.sub.2 for a prolonged time on the injured sites, by preventing the injured sites from external microbial contamination by forming a seal over the injured sites, and by providing oxygen molecule to the injured tissues. Unlike the conventional ointment, the polymeric gel provides a dry, clean, and comfortable coating on the injured sites upon application. Such gel formulations may contain polymeric drug delivery vehicles like hydroxypropyl methylcellulose (HPMC), methylcellulase (Methocel), hydroxyethylcellulose (HEC), hyaluronic acid, and carboxymethylcellulose (CMC), etc.

(19) ii. A Stable Chlorite/Peroxide Gel

(20) The following Formula 3 is a presently preferred formulation of a chlorite/peroxide gel of the present invention:

(21) TABLE-US-00003 FORMULA 3 Sodium Chlorite 0.02%-0.10% Hydrogen Peroxide 0.005%-0.05% Methocel A 2.0% Boric Acid 0.15% Sodium Chloride 0.75% Pluronic F-68/F-127 0.1% HCl or NaOH Adjust pH 7.4 Purified water Q.S. to volume

(22) Any of the preparations of the present invention may be formulated for sustained release of the active components by forming liposomes of the preparing in accordance with well known liposomal forming techniques and/or by adding to the formulation a pharmaceutically acceptable and effective amount (e.g., typically 1-20 percent by weight) of a sustained release component such as a polymer matrix or one or more of the following: a cellulose ester; hydroxymethylpropyl cellulose; methylhydroxyethyl cellulose; hydroxypropyl cellulose; hydroxyethyl cellulose; carboxymethyl cellulose; a salt of a cellulose ester; cellulose acetate; hydroxypropylmethyl cellulose phthalate; methacrylic acid-methyl methacrylate copolymer; methacrylic acid-ethyl acetate copolymer; polyvinylpyrolidone; polyvinyl alcohol; hyaluronic acid; a phospholipid; cholesterol; a phospholipid having a neutral charge; a phospholipid having a negative charge; dipalmytoyl phoshatidyl choline; dipalmytoyl phoshatidyl serine; and, sodium salts thereof.

(23) iii. A Stable Chlorite/Peroxide Ophthalmic Solution

(24) The following Formula 4 is a presently preferred formulation of a chlorite/peroxide contact lens disinfecting solution for use in cleaning contact lenses residing in or out of the eye. The formulation additionally functions as a tear product for lubrication in dry-eye subjects.

(25) TABLE-US-00004 FORMULA 4 Sodium Chlorite 0.002%-0.20% Hydrogen Peroxide 0.005%-0.05% Hyaluronic Acid 0.001%-0.50% Boric Acid 0.15% Sodium Chloride 0.75% Pluronic 127 0.05%-2.0% HCl or NaOH Adjust pH to 7.4 Purified Water Q.S. to Volume

(26) As indicated earlier, the chlorite/peroxide preparation of the present invention, whether it be in the form of liquid solution, gel, ointment, cream, spray, etc., is specifically composed to maintain chlorite such as sodium chlorite and hydrogen peroxide as active ingredients at a pH range of 5.0-8.8 without generating chlorine dioxide during storage at room temperature. By way of illustration, multiple experiments were conducted on the liquid sodium chlorite/hydrogen peroxide solution in accordance with Formula 2 at different levels of pH within the specified range. However, it should be expressly stated herein that such experimentations should in no way be limited to liquid solution forms only, but are performed to illustrate the non-production of chlorine dioxide in the various forms of the present chlorite/peroxide preparation at different pH levels.

(27) The following experimentations were designed to demonstrate the stability of chlorite such as sodium chlorite and hydrogen peroxide antibacterial formulation at neutral, basic and acidic levels of pH. More specifically, the quantitative levels of sodium chlorite and the generation of chlorine dioxide were determined at the pH levels of 7.3, 8.0, 8.8, 7.0, 6.44 and 6.0. 0.1 Normal hydrochloric acid solution and 0.1 Normal sodium hydroxide solution were applied to adjust the pH levels in the experimentations. Sterile 0.9% sodium chloride sterile solution was also applied. A placebo solution with the following formulation was further applied in a spectrophotometer (e.g., Lambda 20 Model UV-Vis. spectrophotometer) to find and measure the levels of sodium chlorite and the generation of chlorine dioxide at varying pH levels:

(28) TABLE-US-00005 Placebo Solution Hydrogen Peroxide 0.02% Carboxymethyl Cellulose 0.01% Boric Acid 0.15% Sodium Chloride 0.75% Pluronic F-68/F-127 0.1% HCl or NaOH Adjust pH 7.3 Purified water Q.S. to volume

Experiment 1: pH Level of 7.3

(29) Experiment: Fill the first cuvette with the placebo solution, wipe it clean, and place the cuvette in the standard beam path of the spectrophotometer. Fill the second cuvette with the liquid sodium chlorite/hydrogen peroxide solution, wipe it clean and place the cuvette in the sample beam path of the spectrophotometer. Scan the solutions from 200 nm to 400 nm and record the results. Plot and printout the results, as illustrated in the graph shown in FIG. 1.

(30) Result: The liquid solution contained sodium chlorite and hydrogen peroxide as active ingredients, as well as buffering and tonicity agents at the pH level of 7.3. The placebo solution contained hydrogen peroxide as active ingredient, as well as buffering and tonicity agents at the pH level of 7.3.

(31) Hydrogen peroxide does not absorb in the 200 nm to 400 nm range. Therefore, as seen in FIG. 1, absorption peaks for hydrogen peroxide were not detected.

(32) Sodium chlorite has an absorption maximum at 260 nm, while chlorine dioxide which is a degradation product of sodium chlorite has an absorption maximum at 355 nm-358 nm.

(33) Scanning the solutions that have a pH of 7.3 between the 200 nm and 400 nm will give a quantitative value for sodium chlorite as well as chlorine dioxide in the same scan.

(34) Interpretation: The liquid sodium chlorite/hydrogen peroxide solution does show sodium chlorite peak at 260 nm, but does not show any chlorine dioxide peak at 355 nm-358 nm.

(35) This clearly indicates that at pH level of 7.3, the liquid sodium chlorite/hydrogen peroxide solution has only sodium chlorite, and does not contain any quantities of chlorine dioxide. This is a clear indication that sodium chlorite is stable at pH level of 7.3, and the sodium chlorite is not breaking up and forming the chlorine dioxide.

Experiment 2: pH Level of 8.0

(36) Experiment: Dispense 25 mL. of the placebo solution and 25 mL. of the liquid sodium chlorite/hydrogen peroxide solution into 2 clean containers. Add 0.1 Normal sodium hydroxide solution to each container so as to adjust the pH of both the placebo solution as well as the liquid solution to a pH level of 8.0.

(37) Fill one of the cuvette with the placebo solution, wipe it clean, and place the cuvette in the standard beam path of the spectrophotometer. Fill the second cuvette with the liquid sodium chlorite/hydrogen peroxide solution, wipe it clean and place the cuvette in the sample beam path of the spectrophotometer. Scan the solutions from 200 nm to 400 nm and record the results. Plot and printout the results, as illustrated in the graph shown in FIG. 2.

(38) Result: The liquid sodium chlorite/hydrogen peroxide solution contained sodium chlorite and hydrogen peroxide as active ingredients, as well as buffering and tonicity agents at the pH level of 8.0. The placebo solution contained hydrogen peroxide as active ingredient, as well as buffering and tonicity agents at the pH level of 8.0.

(39) As mentioned shortly above, hydrogen peroxide does not absorb in the 200 nm to 400 nm range. Therefore, as seen in FIG. 2, absorption peaks for hydrogen peroxide were not detected. As also mentioned above, sodium chlorite has an absorption maximum at 260 nm, while chlorine dioxide which is a degradation product of sodium chlorite has an absorption maximum at 355 nm-358 nm.

(40) Scanning the solutions that have a pH level of 8.0 between the 200 nm and 400 nm will give a quantitative value for sodium chlorite as well as chlorine dioxide in the same scan.

(41) Interpretation: The liquid sodium chlorite/hydrogen peroxide solution does show sodium chlorite peak at 260 nm, but does not show any chlorine dioxide peak at 355 nm-358 nm. This clearly indicates that at the pH level of 8.0, the liquid sodium chlorite/hydrogen peroxide solution has only sodium chlorite, and does not contain any quantities of chlorine dioxide. This is a clear indication that sodium chlorite is stable at the pH level of 8.0, and the chlorite is not breaking up and forming chlorine dioxide.

Experiment 3: pH Level of 8.8

(42) Dispense 25 mL. of the placebo solution and 25 mL. of the liquid sodium chlorite/hydrogen peroxide solution into 2 clean containers. Add 0.1 Normal sodium hydroxide solution to each container so as to adjust the pH of both the placebo solution as well as the liquid solution to a pH level of 8.8.

(43) Fill one of the cuvette with the placebo solution, wipe it clean, and place the cuvette in the standard beam path of the spectrophotometer. Fill the second cuvette with the liquid sodium chlorite/hydrogen peroxide solution, wipe it clean and place the cuvette in the sample beam path of the spectrophotometer. Scan the solutions from 200 nm to 400 nm and record the results. Plot and printout the results, as illustrated in the graph shown in FIG. 3.

(44) Result: The liquid sodium chlorite/hydrogen peroxide solution contained sodium chlorite and hydrogen peroxide as active ingredients, as well as buffering and tonicity agents at the pH level of 8.8. The placebo solution contained hydrogen peroxide as active ingredient, as well as buffering and tonicity agents at the pH level of 8.8.

(45) As already discussed, hydrogen peroxide does not absorb in the 200 nm to 400 nm range. Therefore, as seen in FIG. 3, absorption peaks for hydrogen peroxide were not detected. As also discussed, sodium chlorite has an absorption maximum at 260 nm, while chlorine Dioxide which is a degradation product of sodium chlorite has an absorption maximum at 355 nm-358 nm.

(46) Scanning the solutions that have a pH level of 8.8 between the 200 nm and 400 nm will give a quantitative value for sodium chlorite as well as chlorine dioxide in the same scan.

(47) Interpretation: The liquid sodium chlorite/hydrogen peroxide solution does show sodium chlorite peak at 260 nm, but does not show any chlorine dioxide peak at 355 nm-358 nm. This clearly indicates that at the pH level of 8.8, the liquid sodium chlorite/hydrogen peroxide solution has only sodium chlorite, and does not contain any quantities of chlorine dioxide. This is a clear indication that sodium chlorite is stable at the pH level of 8.8, and the chlorite is not breaking up and forming chlorine dioxide.

Experiment 4: pH Level of 7.0

(48) Experiment: Dispense 25 mL. of the placebo solution and 25 mL. of the liquid sodium chlorite/hydrogen peroxide solution into 2 clean containers. Add 0.1 Normal hydrochloric acid solution to each container so as to adjust the pH of both the placebo solution as well as the liquid solution to a pH level of 7.0.

(49) Fill one of the cuvette with the placebo solution, wipe it clean, and place the cuvette in the standard beam path of the spectrophotometer. Fill the second cuvette with the liquid sodium chlorite/hydrogen peroxide solution, wipe it clean and place the cuvette in the sample beam path of the spectrophotometer. Scan the solutions from 200 nm to 400 nm and record the results. Plot and printout the results, as illustrated in the graph shown in FIG. 4.

(50) Result: The liquid sodium chlorite/hydrogen peroxide solution contained sodium chlorite and hydrogen peroxide as active ingredients, as well as buffering and tonicity agents at the pH level of 7.0. The placebo solution contained hydrogen peroxide as active ingredient, as well as buffering and tonicity agents at the pH level of 7.0. Hydrogen peroxide does not absorb in the 200 nm to 400 nm range. Therefore, as seen in FIG. 4, absorption peaks for hydrogen peroxide were not detected.

(51) Sodium chlorite has an absorption maximum at 260 nm, while chlorine dioxide which is a degradation product of sodium chlorite has an absorption maximum at 355 nm-358 nm. Scanning the solutions that have a pH of 7.0 between the 200 nm and 400 nm will give a quantitative value for sodium chlorite as well as chlorine dioxide in the same scan.

(52) Interpretation: The sodium chlorite/hydrogen peroxide solution does show sodium chlorite peak at 260 nm, but does not show any chlorine dioxide peak at 355 nm-358 nm. This clearly indicates that at the pH level of 7.0, the liquid solution has only sodium chlorite, and does not contain any quantities of chlorine dioxide. This is a clear indication that sodium chlorite is stable at pH of 7.0, and the chlorite is not breaking up and forming chlorine dioxide.

Experiment 5: pH Level of 6.44

(53) Experiment: Dispense 25 mL. of the placebo solution and 25 mL. of the liquid sodium chlorite/hydrogen peroxide solution into 2 clean containers. Add 0.1 Normal hydrochloric acid solution to each container so as to adjust the pH of both the placebo solution as well as the liquid solution to a pH level of 6.44.

(54) Fill one of the cuvette with the placebo solution, wipe it clean, and place the cuvette in the standard beam path of the spectrophotometer. Fill the second cuvette with the liquid solution, wipe it clean and place the cuvette in the sample beam path of the spectrophotometer. Scan the solutions from 200 nm to 400 nm and record the results. Plot and printout the results, as illustrated in the graph shown in FIG. 5.

(55) Result: The liquid sodium chlorite/hydrogen peroxide solution contained sodium chlorite and hydrogen peroxide as active ingredients, as well as buffering and tonicity agents at the pH level of 6.44.

(56) The placebo solution contained hydrogen peroxide as active ingredient, as well as buffering and tonicity agents at pH=6.44. Hydrogen peroxide does not absorb in the 200 nm to 400 nm range, and thus no absorption peaks for hydrogen peroxide were detected. Sodium chlorite has an absorption maximum at 260 nm, while chlorine dioxide which is a degradation product of sodium chlorite has an absorption maximum at 355 nm-358 nm.

(57) Scanning the solutions that have a pH of 6.44 between the 200 nm and 400 nm will give a quantitative value for sodium chlorite as well as chlorine dioxide in the same scan.

(58) Interpretation: The liquid sodium chlorite/hydrogen peroxide solution does show sodium chlorite peak at 260 nm, but does not show any chlorine dioxide peak at 355 nm-358 nm. This clearly indicates that at pH of 6.44, the liquid solution has only sodium chlorite, and does not contain any quantities of chlorine dioxide. This is a clear indication that sodium chlorite is stable at pH of 6.44, and the chlorite is not breaking up and forming chlorine dioxide.

Experiment 6: pH Level of 6.0

(59) Experiment: Dispense 25 mL. of the placebo solution and 25 mL. of the liquid sodium chlorite/hydrogen peroxide solution into 2 clean containers. Add 0.1 Normal hydrochloric acid solution to each container so as to adjust the pH of both the placebo solution as well as the liquid solution to a pH level of 6.0.

(60) Fill one of the cuvette with the placebo solution, wipe it clean, and place the cuvette in the standard beam path of the spectrophotometer. Fill the second cuvette with the liquid sodium chlorite/hydrogen peroxide solution, wipe it clean and place the cuvette in the sample beam path of the spectrophotometer. Scan the solutions from 200 nm to 400 nm and record the results. Plot and printout the results, as illustrated in the graph shown in FIG. 6.

(61) Result: The liquid sodium chlorite/hydrogen peroxide solution contained sodium chlorite and hydrogen peroxide as active ingredients, as well as buffering and tonicity agents at the pH level of 6.0. The placebo solution contained hydrogen peroxide as active ingredient, as well as buffering and tonicity agents at the pH level of 6.0. Hydrogen peroxide does not absorb in the 200 nm to 400 nm range. Therefore, as seen in FIG. 6, absorption peaks for hydrogen peroxide were not detected.

(62) Sodium chlorite has an absorption maximum at 260 nm, while chlorine dioxide which is a degradation product of sodium chlorite has an absorption maximum at 355 nm-358 nm. Scanning the solutions that have a pH of 6.0 between the 200 nm and 400 nm will give a quantitative value for sodium chlorite as well as chlorine dioxide in the same scan.

(63) Interpretation: The sodium chlorite/hydrogen peroxide solution does show sodium chlorite peak at 260 nm, but does not show any chlorine dioxide peak at 355 nm-358 nm. This clearly indicates that at pH level of 6.0, the liquid solution has only sodium chlorite, and does not contain any quantities of chlorine dioxide. This is a clear indication that sodium chlorite is stable at pH of 6.0, and the chlorite is not breaking up and forming chlorine dioxide.

Experiment 7: pH Level of 1.5

(64) Experiment: Dispense 25 mL. of the placebo solution and 25 mL. of the liquid sodium chlorite/hydrogen peroxide solution into 2 clean containers. Add 0.1 Normal hydrochloric acid solution to each container so as to adjust the pH of both the placebo solution as well as the bactericidal solution to a pH of 1.5.

(65) Fill one of the cuvette with the placebo solution, wipe it clean, and place the cuvette in the standard beam path of the spectrophotometer. Fill the second cuvette with the liquid solution, wipe it clean and place the cuvette in the sample beam path of the spectrophotometer. Scan the solutions from 200 nm to 400 nm and record the results. Plot and printout the results, as illustrated in the graph shown in FIG. 7.

(66) Result: The liquid sodium chlorite/hydrogen peroxide solution contained sodium chlorite and hydrogen peroxide as active ingredients, as well as buffering and tonicity agents at pH of 1.5. The placebo solution contained hydrogen peroxide as active ingredient, as well as buffering and tonicity agents at pH of 1.5. As explained earlier, hydrogen peroxide does not absorb in the 200 nm to 400 nm range, and as such, no absorption peaks for hydrogen peroxide were detected.

(67) Also explained earlier, sodium chlorite has an absorption maximum at 260 nm, while chlorine dioxide which is a degradation product of sodium chlorite has an absorption maximum at 355 nm-358 nm. Scanning the solutions that have a pH of 1.5 between the 200 nm and 400 nm will give a quantitative value for sodium chlorite as well as chlorine dioxide in the same scan.

(68) Interpretation: The liquid sodium chlorite/hydrogen peroxide solution does not show sodium chlorite peak at 260 nm, but does show a large chlorine dioxide peak at 355 nm-358 nm. This clearly indicates that at the pH level of 1.5, the liquid sodium chlorite/hydrogen peroxide solution does not have any sodium chlorite. Rather, it clearly shows that the sodium chlorite has been degraded and converted to chlorine dioxide. This is a clear indication that at pH of 1.5, sodium chlorite is very unstable, and all chlorite that is present in the liquid solution is converted to chlorine dioxide.

Results for Experiments 1-7

(69) The liquid sodium chlorite/hydrogen peroxide solution contained sodium chlorite and hydrogen peroxide as active ingredients, as well as buffering and tonicity agents at the pH levels of 1.5, 6.0, 6.44, 7.0, 7.3, 8.0 and 8.8.

(70) The placebo solution contained hydrogen peroxide as active ingredient, as well as buffering and tonicity agents at the pH levels of 1.5, 6.0, 6.44, 7.0, 7.3, 8.0 and 8.8.

(71) Hydrogen peroxide does not absorb in the 200 nm to 400 nm range.

(72) Sodium chlorite has an absorption maximum at 260 nm, while chlorine dioxide has an absorption maximum at 355 nm-358 nm.

(73) Scanning the solutions between the 200 nm and 400 nm gave a quantitative value for sodium chlorite as well as chlorine dioxide in the same scan.

Interpretation of Results for Experiments 1-7

(74) The liquid sodium chlorite/hydrogen peroxide solutions at the pH levels of 6.0, 6.44, 7.0, 7.3, 8.0 and 8.8 does show the presence of sodium chlorite peak at 260 nm, but does not show the presence of chlorine dioxide peak at 355 nm-358 nm.

(75) In contrast, the liquid sodium chlorite/hydrogen peroxide solution at pH of 1.5 does not show the presence of sodium chlorite peak at 260 nm, but does show the presence of chlorine dioxide peak at 355 nm-358 nm.

Conclusion of Results for Experiments 1-7

(76) The results clearly show that one can quantitatively determine the level of sodium chlorite as well as chlorine dioxide which is present in the liquid sodium chlorite/hydrogen peroxide solution at the pH levels of 1.5, 6.0, 6.44, 7.0, 7.3, 8.0 and 8.8.

(77) The results also show that the storage of the liquid sodium chlorite/hydrogen peroxide solution at about room temperature (e.g., in a white opaque bottle exposed to air at room temperature) does not produce any chlorine dioxide as determined by the absence of any absorbance at 355 nm-358 nm.

(78) In conclusion, the results of Experiments 1-7 clearly indicate that the liquid sodium chlorite/hydrogen peroxide solution retains sodium chlorite at the pH range of 6.0-8.8 without the generation of chlorine dioxide. The liquid solution, however, degrades and generates chlorine dioxide upon the acidification of the solution to pH of 1.5. Thus, these results also strongly indicate that the liquid sodium chlorite/hydrogen peroxide solution does not contain chlorine dioxide when it is manufactured, nor does the solution degrade to generate chlorine dioxide after storage at about room temperature at the pH levels of 6.0, 6.44, 7.0, 7.3, 8.0, or 8.8.

(79) Furthermore, these results present clear evidence that the liquid sodium chlorite/hydrogen peroxide solution of the present invention has its bactericidal properties in the pH range studied due to the sodium chlorite/hydrogen peroxide and not due to chlorine dioxide. This is very much unlike other prior art inventions that have sodium chlorite as a starting material as, but the active bactericide is the chlorine dioxide which is generated by the acidification of the sodium chlorite.

B. Examples of Therapeutic Applications

(80) The following are specific examples of therapeutic applications of the chlorite/peroxide preparations of the present invention.

i. Example 1: Treatment of PsoriasisNo Crossover

(81) A human patient having psoriasis plaques present on both arms is treated as follows: Twice daily application to plaques on the left arm only, of a chlorite/peroxide solution having the following formulation:

(82) TABLE-US-00006 Sodium Chlorite 0.06% Hydrogen Peroxide 0.01% HPMC 2.0% Boric Acid 0.15% HCl or NaOH to adjust pH 7.4 Purified water Q.S. to volume

(83) Twice daily application to plaques on the right arm only of a commercially available 0.1% triamcinolone acetonide cream.

(84) The chlorite/peroxide treated psoriatic plaques on the right arm began to become less severe within 24 hours of beginning treatment and had substantially disappeared within three days of beginning treatment. However, the triamcinolone acetonide treated psoriatic plaques present on the left arm remained unchanged and inflamed during the two week treatment period.

ii. Example 2: Treatment of PsoriasisCrossover

(85) A human patient having psoriasis plaques present on both arms is treated for two weeks, as follows: Twice daily application to plaques on the left arm only, of a chlorite/peroxide solution having the following formulation:

(86) TABLE-US-00007 Sodium Chlorite 0.06% Hydrogen Peroxide 0.01% HPMC 2.0% Boric Acid 0.15% HCl or NaOH to adjust pH 7.4 Purified water Q.S. to volume/100%

(87) Twice daily application to plaques on the right arm only of a commercially available 0.1% triamcinolone acetonide cream.

(88) The chlorite/peroxide treated psoriatic plaques on the right arm began to become less severe within 24 hours of beginning treatment and had substantially disappeared within one week of beginning treatment. However, the triamcinolone acetonide treated psoriatic plaques present on the left arm remained unchanged and inflamed during the two week treatment period.

(89) Beginning the day after the end of the initial two week treatment period, and continuing for a second two week treatment period, the patient was treated as follows:

(90) Twice daily application to plaques on the left arm only of the same commercially available 0.1% triamcinolone acetonide cream described hereabove in this example.

(91) Twice daily application to plaques on the right arm only, of the same chlorite/peroxide sustained release gel described hereabove in this example.

(92) Within 24 hours of commencing the second treatment period, the psoriatic lesions on the right arm began to subside. By day three and continuing through the end of the second two week treatment period, the psoriatic lesions on the right arm had substantially disappeared.

iii. Example 3: Treatment of Cold Sores

(93) A patient with painful, fluid-containing cold sores (i.e., chancre sores) on his lips was treated twice daily by application to the lips of a chlorite/peroxide preparation prepared in accordance with Formula 1 above.

(94) Within 6 to 12 hours of the first application of the chlorite/peroxide preparation, the patient reported that the pain had subsided. Within 24 hours of the first application of the chlorite/peroxide preparation, the fluid contained within the cold sores had substantially dissipated and the cold sores appeared dry. Within six days of the first application of the chlorite/peroxide preparation the cold sores had substantially disappeared and the lips appeared normal, whereas cold sores of such severity typically require substantially longer than six days to completely disappear and heal.

iv. Example 4: Treatment of Venous Ulcer

(95) A patient with a venous ulcer on the right leg of 3-4 cm diameter which had been present for 9-12 months was treated by twice daily application to the ulcer of gauze soaked with a chlorite/peroxide liquid solution prepared in accordance with Formula 1 above.

(96) Within three days after commencement of treatment the ulcer appeared clean and dry. Within 14 days of the commencement of treatment the ulcer began to decrease in size and healthy new tissue was observed about its periphery. At 35 days after commencement of treatment, the ulcer had completely healed, without scarring, and the area where the ulcer had been located was free of pain.

v. Example 5: Treatment of Diabetic Decubitus Ulcer

(97) A non-ambulatory, diabetic patient with decubitus ulcers on both legs and some toes, of 12-18 month duration, was treated by daily application of clean, sterile gauze to the ulcers and saturation of each gauze, three times each day, with a liquid chlorite/peroxide solution prepared in accordance with Formula 1 above. Within four to seven days of commencing the chlorite/hydrogen peroxide treatments the ulcers began to appear less inflamed, clean and dry. About seven to ten days after commencement of the chlorite/hydrogen peroxide treatment, granulation tissue began to form within the ulcers. Within 12 to 14 days, re-epithelialization was observed to have begun within the ulcerated areas except for one toe ulcer which had been particularly severe and had permeated to the bone of the toe. Within 30 to 45 days of the commencement of treatment, all of the ulcers except for the severe toe ulcer had completely closed and re-epithelialized, without irregular scar formation. Also, at 30 to 45 days after the commencement of treatment, the toe ulcer had also become substantially smaller (but was not completely closed) and the patient was able to walk. The liquid and or gel formulations of the present invention, such as Formulas 1 and 2 above, may also be applied topically to prevent scar formation due to wounds, burns, acne, infections, trauma, surgical incision, or any other scar-forming lesion or disorder.

vi. Example 6

(98) a. Treatment of Dry Eye Conditions

(99) Subjects with dry eye conditions have itchy and scratchy eyes. In extreme cases, the subjects have more serious problems that can interfere with health maintenance. Subjects were treated with a preferred tear product of the following formulation:

(100) TABLE-US-00008 Sodium Chlorite 0.005%-0.02% Hydrogen Peroxide 0.01% Methylcellulose A4M 0.075% Hyaluronic Acid 0.10%-0.125% Boric Acid 0.15% Sodium Chloride, USP 0.75% Pluronic 127 0.10% HCl or NaOH Adjust pH to 7.4 Purified Water Q.S. to Volume

(101) Testing of dry eye subjects with rose bengal stain or fluorescein gives a good indication regarding the condition of the corneal epithelial health, while rose bengal staining provides a good indication of the number of dead epithelial cells on the cornea as well as conjunctiva.

(102) Two subjects with dry eye condition were tested with rose bengal stain, and the quantitative staining to the cornea and conjunctiva was documented by photographs. The subjects started using the above preferred tear product at a dosage of two drops three times per day. At the end of two weeks, the two subjects were tested with rose bengal stain and the level of staining was quantitatively documented by photography. The results showed a 50% to 70% reduction in rose bengal staining, which clearly indicates that the preferred tear formulation was ameliorating the corneal and conjunctival cells from dying.

(103) In addition to an objective determination of the health of the epithelial cells, the two subjects were tested subjectively regarding the safety and efficacy of the preferred tear product. First of all, slit-lamp biomicroscopy of the subjects during the two-week treatment period did not show any redness, irritation, inflammation, or other signs of discomfort. Second, the subjects indicated that the application of the tear product completely removed symptoms of redness, itching, scratching, pain, and dryness due to dry eye while providing lubrication that lasted for several hours. It is therefore evident that the tear product exhibits both safety and efficacy in the treatment of dry eye. As is further recognized in view of the foregoing antimicrobial activity of such compositions, the tear product will also have efficacy in enhancing wound healing within the eye such as after surgery where bacterial infections are to be avoided.

(104) b. Treatment of Allergic Conjunctivitis

(105) In addition to treating dry eye condition with the above preferred tear product, the product was also tested in the treatment of conditions from allergic conjunctivitis. In particular, two subjects suffering from allergic conjunctivitis including itchy, scratchy eyes with constant tearing applied two drops of the product three times per day. This dosage resulted in the disappearance of the symptoms.

C. Examples of Contact Lens Cleansing

i. Example 1: Soaking, Cleaning and Disinfecting

(106) The following formulation is a preferred disinfecting solution applicable to the cleaning of contact lenses by conventional soaking.

(107) TABLE-US-00009 Sodium Chlorite 0.05% Hydrogen Peroxide 0.02% Methylcellulose A4M 0.075% Hyaluronic Acid 0.05%-0.10% Boric Acid 0.15% Pluronic 127 0.25%-0.50% Sodium Chloride USP 0.75% HCl or NaOH Adjust pH to 7.4 Purified Water Q.S. to Volume

(108) Six subjects using soft hydrophilic contact lenses soaked the lenses in the above disinfecting solution and then placed the lenses directly into the eyes. Soaking was performed nightly or on an as-needed basis. All six subjects reported that the lenses felt very comfortable, and that no adverse effects (e.g., burning, stinging, redness, pain) were experienced. Additionally, the solution extended the comfort and clean condition of the lenses for several weeks beyond such extension experienced with other commercially available disinfecting solutions.

(109) The disinfecting solution can be used with soft hydrophilic lenses of varying water content (e.g., 38% to 75%), as well as with silicone acrylate rigid gas permeable lenses. Cycling studies of soft lenses soaked daily in the solution for 30 days showed no damage or change in the physical and chemical characteristics of the lenses. Eye comfort, as earlier noted, is achieved through non-binding and non-accumulating of preservative in soft or rigid gas permeable lenses, while such binding and accumulation can be found in certain currently commercially available formulations to cause irritation and discomfort.

ii. Example 2: Cleaning while Wearing

(110) The following formulation is a preferred disinfecting in-eye solution applicable to the cleaning of contact lenses while they are being worn by introducing the solution into the eye:

(111) TABLE-US-00010 Sodium Chlorite 0.02% Hydrogen Peroxide 0.01%-0.02% Methylcellulose A4M 0.075% Hyaluronic Acid 0.075%-0.10% Boric Acid 0.15% Sodium Chloride USP 0.75% Pluronic 127 0.75% HCl or NaOH Adjust pH to 7.4 Purified Water Q.S. to Volume

(112) Four subjects applied two drops of the above in-eye solution three times per day for 30 days to contact lenses while being worn. Examinations of all of the subjects showed no irritation, burning, stinging, or adverse effects of any kind. These subjects further reported that the solution felt soothing and lubricating.

(113) Two subjects were involved in a comparative study where, first of all, they wore ACUVUE disposable lenses continuously for two weeks with occasional removal and cleaning with commercially available cleaning solutions followed with a saline rinse. After 14 days, the lenses became very gritty and uncomfortable, and were discarded. Second, the two subjects started with new ACUVUE lenses and practiced daily application of the present in-eye solution three times per day without removing or touching the lenses. These subjects were able to wear the lenses for three to four weeks before replacement. Additionally, the inconvenience of cleaning the lenses outside the eye was completely eliminated, as was the risk of lens loss, tearing, or contamination. It is therefore evident that the present in-eye cleaning solution provides cleansing efficacy as well as convenience.

D. In-Vitro and In-Vivo Antimicrobial Efficacy

(114) i. Synergistic Activity

(115) Tables I and II compare the antimicrobial effects of (a) 400 ppm sodium chlorite alone; (b) 200 ppm hydrogen peroxide alone; and (c) 400 ppm sodium chlorite and 200 ppm hydrogen peroxide in combination against antibiotic-resistant strains of staphylococcus haemolyticus (Table I) and pseudomonas aeruginosa (Table II) both isolated from human infected eyes. Tables I and II summarize the antimicrobial effects observed at time points one and two hours after introduction of the test solutions.

(116) TABLE-US-00011 TABLE I (staphylococcus haemolyticus: Initial inoculum = 1.01 10.sup.7: Log 7.03) Log Reduction Log Reduction Time NaClO.sub.2 alone H.sub.20.sub.2 alone NaClO.sub.2 & H.sub.20.sub.2 (hours) (400 ppm) (200 ppm) (400 ppm & 200 ppm) 1 0.11 0.20 0.69 2 1.01 0.23 2.43

(117) TABLE-US-00012 TABLE II (pseudomonas aeruginosa: Initial inoculum = 2.22 10.sup.6: Log 6.35) Log Reduction Log Reduction Time NaClO.sub.2 alone H.sub.20.sub.2 alone NaClO.sub.2 & H.sub.20.sub.2 (hours) (400 ppm) (200 ppm) (400 ppm & 200 ppm) 1 0.351 0.01 0.04 2 1.35 0.54 6.35

(118) In the experiment summarized in Table I, sodium chlorite alone caused a Log reduction in staphylococcus haemolyticus bacteria of 0.11 at 1 hour and 1.01 at 2 hours. Hydrogen peroxide alone caused a Log reduction in staphylococcus haemolyticus bacteria of 0.20 at 1 hour and 0.23 at 2 hours and the combination of sodium chlorite and hydrogen peroxide caused a Log reduction in staphylococcus haemolyticus bacteria of 0.69 at 1 hour and 2.43 at 2 hours. Thus, in this experiment, the antimicrobial effect of the sodium chlorite-hydrogen peroxide combination was significantly greater than the sums of the effects of the sodium chlorite and hydrogen peroxide alone, at least at the 2 hour time point. Accordingly, it is concluded that the sodium chlorite-hydrogen peroxide combination exhibited a supra-additive effect against the strain of staphylococcus haemolyticus used in this experiment.

(119) In the experiment summarized in Table II, sodium chlorite along caused a Log reduction in pseudomonas aeruginosa bacteria of 0.35 at 1 hour and 1.35 at 2 hours. Hydrogen peroxide alone caused a Log reduction in pseudomonas aeruginosa bacteria of 0.01 at 1 hour and 0.54 at 2 hours and the combination of sodium chlorite and hydrogen peroxide caused a Log reduction in pseudomonas aeruginosa bacteria 0.04 at 1 hour and 6.35 at 2 hours. Thus, in this experiment, the antimicrobial effect of the sodium chlorite-hydrogen peroxide combination was significantly greater than the sums of the effects of the sodium chlorite and hydrogen peroxide alone, at least in the 2 hour time point. Accordingly, it is concluded that the sodium chlorite-hydrogen peroxide combination exhibited a supra-additive effect against the strain of pseudomonas aeruginosa used in this experiment.

(120) ii. Animal Testing

(121) S. haemolyticus keratitus was induced in respective right eyes of 12 rabbits by dropping broth containing 50,000 CFU/ml of S. haemolyticus onto abraded corneas of these eyes. After 24 hours, all corneas were likewise infected, and the rabbits were divided randomly into three groups. The rabbits (five) of Group I then were treated with the chlorite-hydrogen peroxide formulation defined above as cleaning while wearing contact lenses (here termed Bactericide); the rabbits (five) of Group II were treated with commercially available 0.3% ofloxacin antibiotic ophthalmic solution; and the rabbits (two) of Group III were untreated to serve as a control.

(122) At 24 and 48 hours post infection, the rabbits underwent visual eye examination, photographic documentation and biomicroscopy. After 24 hours of treatment, three animals each from Groups I and II and one animal from Group III were sacrificed. The eyes were enucleated and an 8 mm disc of cornea was homogenized and plated onto growth media for microbial isolation and quantification. After 48 hours of treatment, the same procedure was followed for the remaining animals.

(123) Tables III, IV and V summarize the results of this experimentation. As is there apparent, the Bactericide of the present invention exhibited superior overall results as compared to the competing commercially available regimens. The results therefore confirm that the clinical efficacy of the Bactericide is better than the antibiotic treatment. In addition to having excellent bactericidal properties, it is demonstrated that bactericide superiority is probably attributable to inactivation of bacterial proteolytic enzymes (thus decreasing bacterial virulence) and inactivation of bacterial toxins responsible for inflammation and hyperemia.

(124) TABLE-US-00013 TABLE III IN-VIVO ANTIMICROBIAL EFFICACY IN INFECTIOUS S. HAEMOLYTICUS KERATITIS IN RABBITS Post Treatment Group I Group II Group III Time Bactericide 0.3% Ofloxacin Untreated Control 24 hours i) 0 CFU i) 23,000 CFU ii) 18,000 CFU ii) 5,000 CFU iii) 0 CFU iii) 11,000 CFU 39,000 CFU Average 6,000 CFU 13,000 CFU 39,000 CFU 48 hours i) 0 CFU i) 5,000 CFU ii) 0 CFU ii) 5,200 CFU 231,000 CFU Average 0 CFU 5,100 CFU 231,000 CFU

(125) TABLE-US-00014 TABLE IV IN-VIVO CLINICAL EFFICACY IN INFECTIOUS S. HAEMOLYTICUS KERATITIS IN RABBITS Group I Group II Group III Time Bactericide 0.3% Ofloxacin Untreated Control 24 hours inflammation (+2) inflammation(+2) inflammation(+2) after hyperemia (+2) hyperemia (+2) hyperemia (+2) infection corneal edema corneal edema corneal edema (+2) (+2) (+2) 24 hours inflammation (0) inflammation(+2) inflammation(+3) after hyperemia (0) hyperemia (+2) hyperemia (+3) treatment corneal edema (0) corneal edema corneal edema (+2) (+3) 48 hours inflammation (0) inflammation(+1) inflammation(+3) after hyperemia (0) hyperemia (+1) hyperemia (+3) treatment corneal edema (0) corneal edema corneal edema (+1) (+3)

(126) TABLE-US-00015 TABLE V IN-VITRO INHIBITION OF PROTEOLYTIC ENZYME ACTIVITY Inhibition of proteolytic enzyme activity of Trypsin and porcine pancreatic Elastase Concentration of % Inhibition of Enzyme Bactericide Enzyme activity Elastase (porcine) 0.18 ppm 46% Trypsin 0.12 ppm 28%

E. Ocular Tolerability and Degradation Speed

(127) i. Ocular Tolerability of High Levels of Hydrogen Peroxide

(128) Previously, it was believed that the upper limit of human ocular tolerability of hydrogen peroxide is about 100 ppm (0.01 wt. %). See Paugh, J. R., Brennan, N. A., and Efron, N., Ocular Response to Hydrogen Peroxide, Am. J. Optom. Physiol. Opt. 1988 February; 65(2):91-8. The following experiments show, however, that when combined with sodium chlorite, hydrogen peroxide is well tolerated by human eyes in levels up to 500 ppm (0.05 wt. %). In all of the following experiments, the hydrogen peroxide was combined with 400 ppm (0.04 wt. %) of sodium chlorite in 0.2% boric acid at pH 7.4 and filtered through a 0.2 m Acrodisc syringe filter. Two drops of each formulation were then placed in the cul-de-sac of two normal human eyes. Upon instillation of the drops, the subjects were instructed to close their eyelids. The subjects' ocular symptoms of the treated eyes were observed and graded over a period of one hour post instillation, for burning and stinging sensation, pain, redness, tearing, itching, photopsia, photophobia, discharge, and foreign body sensation. The observations are presented below.

(129) TABLE-US-00016 TABLE VI Experiment 1: Human Ocular Response to 100 ppm of Hydrogen Peroxide Time post instillation Zero 30 1 2 3 5 10 60 time seconds minute minutes minutes minutes minutes minutes Burning/Stinging 0 0 0 0 0 0 0 0 Pain 0 0 0 0 0 0 0 0 Redness 0 0 0 0 0 0 0 0 Tearing 0 0 0 0 0 0 0 0 Itching 0 0 0 0 0 0 0 0 Photopsia 0 0 0 0 0 0 0 0 Photophobia 0 0 0 0 0 0 0 0 Discharge 0 0 0 0 0 0 0 0 Foreign body 0 0 0 0 0 0 0 0 sensation Grading Scale: 0 = None; +0.5 = Trace; +1 = Mild; +2 = Moderate; +3 = Moderately Severe; +4 = Severe

(130) TABLE-US-00017 TABLE VII Experiment 2: Human Ocular Response to 200 ppm of Hydrogen Peroxide Time post instillation Zero 30 1 2 3 5 10 60 time seconds minute minutes minutes minutes minutes minutes Burning/Stinging 0 0 0 0 0 0 0 0 Pain 0 0 0 0 0 0 0 0 Redness 0 0 0 0 0 0 0 0 Tearing 0 0 0 0 0 0 0 0 Itching 0 0 0 0 0 0 0 0 Photopsia 0 0 0 0 0 0 0 0 Photophobia 0 0 0 0 0 0 0 0 Discharge 0 0 0 0 0 0 0 0 Foreign body 0 0 0 0 0 0 0 0 sensation Grading Scale: 0 = None; +0.5 = Trace; +1 = Mild; +2 = Moderate; +3 = Moderately Severe; +4 = Severe

(131) TABLE-US-00018 TABLE VIII Experiment 3: Human Ocular Response to 300 ppm of Hydrogen Peroxide Time post instillation Zero 30 1 2 3 5 10 60 time seconds minute minutes minutes minutes minutes minutes Burning/Stinging 0 0 0 0 0 0 0 0 Pain 0 0 0 0 0 0 0 0 Redness 0 0 0 0 0 0 0 0 Tearing 0 0 0 0 0 0 0 0 Itching 0 0 0 0 0 0 0 0 Photopsia 0 0 0 0 0 0 0 0 Photophobia 0 0 0 0 0 0 0 0 Discharge 0 0 0 0 0 0 0 0 Foreign body 0 0 0 0 0 0 0 0 sensation Grading Scale: 0 = None; +0.5 = Trace; +1 = Mild; +2 = Moderate; +3 = Moderately Severe; +4 = Severe

(132) TABLE-US-00019 TABLE IX Experiment 4: Human Ocular Response to 400 ppm of Hydrogen Peroxide Time post instillation Zero 30 1 2 3 5 10 60 time seconds minute minutes minutes minutes minutes minutes Burning/Stinging 0 0 0 0 0 0 0 0 Pain 0 0 0 0 0 0 0 0 Redness 0 0 0 0 0 0 0 0 Tearing 0 0 0 0 0 0 0 0 Itching 0 0 0 0 0 0 0 0 Photopsia 0 0 0 0 0 0 0 0 Photophobia 0 0 0 0 0 0 0 0 Discharge 0 0 0 0 0 0 0 0 Foreign body 0 0 0 0 0 0 0 0 sensation Grading Scale: 0 = None; +0.5 = Trace; +1 = Mild; +2 = Moderate; +3 = Moderately Severe; +4 = Severe

(133) TABLE-US-00020 TABLE X Experiment 5: Human Ocular Response to 500 ppm of Hydrogen Peroxide Time post instillation Zero 30 1 2 3 5 10 60 time seconds minute minutes minutes minutes minutes minutes Burning/Stinging 0 0 0 0 0 0 0 0 Pain 0 0 0 0 0 0 0 0 Redness 0 0 0 0 0 0 0 0 Tearing 0 0 0 0 0 0 0 0 Itching 0 0 0 0 0 0 0 0 Photopsia 0 0 0 0 0 0 0 0 Photophobia 0 0 0 0 0 0 0 0 Discharge 0 0 0 0 0 0 0 0 Foreign body 0 0 0 0 0 0 0 0 sensation Grading Scale: 0 = None; +0.5 = Trace; +1 = Mild; +2 = Moderate; +3 = Moderately Severe; +4 = Severe

(134) As can be seen from the above data, hydrogen peroxide levels up to 500 ppm are very well tolerated by human eyes when in the presence of sodium chlorite. There were no signs of irritation, inflammation, or any other adverse effects associated with the instillation of the formulations containing up to 500 ppm hydrogen peroxide. These results show that hydrogen peroxide up to 500 ppm can be very safe and free of any adverse effects to the human eye when used in conjunction with sodium chlorite. Furthermore as discussed above, an outstanding synergistic antimicrobial activity has been discovered with formulations containing sodium chlorite and hydrogen peroxide. Because the previous literature (See Paugh) reported that human ocular tolerability of hydrogen peroxide is about 100 ppm, it is believed that the sodium chlorite must be stabilizing the hydrogen peroxide by forming a kind of transient complex molecule (e.g., peroxychlorite), which exhibits the excellent synergistic antimicrobial activity and degrades to innocuous products like water, oxygen, and salt upon contact with biological systems, as will be discussed in greater detail below.

(135) ii. Hydrogen Peroxide/Sodium Chlorite Degradation in the Eye

(136) The following experiments were designed to determine the speed of self degradation of the hydrogen peroxide/sodium chlorite formulation when placed in the human eye and to determine the level of ocular symptomatology associated with the formulation when used in an in the eye contact lens cleaner product or an artificial tear product.

Experiment 1: in the Eye Contact Lens Cleaner

(137) An in the eye contact lens cleaner containing 0.5 g carboxymethylcellulose, 0.5 g pluronic, and 0.05 g hydrogen peroxide/sodium chlorite mixture in 100 mL sterile water was provided. The cleaner contained 400 ppm sodium chlorite and 100 ppm hydrogen peroxide for a total of 500 ppm hydrogen peroxide/sodium chlorite mixture. Two drops of the cleaner were placed in the cul-de-sac of two normal human eyes. Upon instillation of the drops, the subjects closed their eyelids and pressed their index finger on the medial cantus, so as to block the puncta and stop the tears going into the lachrymal duct.

(138) At 30 second, 1 minute, 2 minute, and 3 minute intervals, the subjects' tear samples were obtained by placing a fresh peroxide test strip in the cul-de-sac of the subjects' eyes. The used peroxide test strips were removed from the eye and left to dry at room temperature for 15 minutes. At the completion of the drying period, the level of hydrogen peroxide/sodium chlorite material left in the tear was estimated by comparing the color formed on the peroxide test strip to that of a standard color chart and recorded as shown below.

(139) TABLE-US-00021 TABLE XI Time post instillation Zero time 30 seconds 1 minute 2 minutes 3 minutes Level of 500 ppm >25 ppm 10 ppm 2 ppm 0.5 ppm hydrogen peroxide/ sodium chlorite

(140) The data presented above shows a rapid reduction in the level of hydrogen peroxide/sodium chlorite in the tear film of the treated subjects. The placing of the index finger on the medial cantus blocks the puncta and does not allow the tears of the subjects to escape into the lachrymal duct. In addition, the closing off the eyelids stops the blinking process and thus stops the pumping action of the tear removal from the treated eyes. As such, it would appear that the rapid reduction in the level of hydrogen peroxide/sodium chlorite from the tears is not due to the loss of the tears of the subjects into the lachrymal duct. Rather, it is believed that the reduction is due to the presence of catalase and superoxide desmutase enzymes in the tears of human subjects. As the drops are placed in the eye of the patients, the catalase and other enzymes start the rapid enzymatic degradation of the hydrogen peroxide/sodium chlorite preparation, whereby in a matter of 3 minutes the level in the tears of the treated subjects is almost undetectable. The results of this experiment tend to show that upon instillation in the eye, the hydrogen peroxide/sodium chlorite mixture behaves like a self destructing preservative with the end products being water, oxygen, and sodium chloride.

(141) Additionally, the ocular symptoms of the treated eyes were observed and graded over a period of one hour post instillation for burning and stinging sensations, pain, redness, tearing, itching, photopsia, photophobia, discharge and for foreign body sensation as shown below.

(142) TABLE-US-00022 TABLE XII Time post instillation Zero 30 1 2 3 5 10 15 30 60 time seconds minute minutes minutes minutes minutes minutes minutes minutes Burning/ 0 0 0 0 0 0 0 0 0 0 Stinging Pain 0 0 0 0 0 0 0 0 0 0 Redness 0 0 0 0 0 0 0 0 0 0 Tearing 0 0 0 0 0 0 0 0 0 0 Itching 0 0 0 0 0 0 0 0 0 0 Photopsia 0 0 0 0 0 0 0 0 0 0 Photophobia 0 0 0 0 0 0 0 0 0 0 Discharge 0 0 0 0 0 0 0 0 0 0 Foreign 0 +0.5 0 0 0 0 0 0 0 0 Body Sensation Grading Scale: 0 = None; +0.5 = Trace; +1 = Mild; +2 = Moderate; +3 = Moderately Severe; +4 = Severe

(143) The above results show that the hydrogen peroxide/sodium chlorite mixture is very well tolerated by the human eye without presenting any signs of irritation, inflammation, or any other adverse effects.

Experiment 2: Artificial Tear Product

(144) An artificial tear product containing 0.15 g sodium hyaluronate, 0.50 g protector, and 0.06 g hydrogen peroxide/sodium chlorite mixture in 100 mL sterile water was provided. The artificial tear product contained 400 ppm sodium chlorite and 200 ppm hydrogen peroxide for a total of 600 ppm hydrogen peroxide/sodium chlorite mixture. Two drops of the cleaner were placed in the cul-de-sac of six normal human eyes. Upon instillation of the drops, the subjects closed their eyelids and pressed their index finger on the medial cantus, so as to block the puncta and stop the tears going into the lachrymal duct.

(145) At zero second, 5 second, 20 second, 30 second, 60 second, 90 second, 120 second, and 180 second intervals, the subjects' tear samples were obtained by placing a fresh peroxide test strip in the cul-de-sac of the subjects' eyes. The used peroxide test strips were removed from the eye and left to dry at room temperature for 15 minutes. At the completion of the drying period, the level of hydrogen peroxide/sodium chlorite material left in the tear was estimated by comparing the color formed on the peroxide test strip to that of a standard color chart and recorded as shown below.

(146) TABLE-US-00023 TABLE XIII Time post instillation Time 5 20 30 60 90 120 180 0 seconds seconds seconds seconds seconds seconds seconds Subject 1 150 ppm 60 ppm 45 ppm 38 ppm Subject 2 75 ppm 38 ppm 38 ppm 23 ppm Subject 3 45 ppm 30 ppm 8 ppm 15 ppm Subject 4 60 ppm 15 ppm 11 ppm 15 ppm Subject 5 60 ppm 23 ppm 5 ppm Subject 6 150 ppm 75 ppm 30 ppm 17 ppm Average 600 ppm 375 ppm 105 ppm 58 ppm 40 ppm 30 ppm 15 ppm 12 ppm

(147) The data presented above shows a rapid reduction in the level of hydrogen peroxide/sodium chlorite in the tear film of the treated subjects. The placing of the index finger on the medial cantus blocks the puncta and does not allow the tears of the subjects to escape into the lachrymal duct. In addition, the closing off the eyelids stops the blinking process and thus stops the pumping action of the tear removal from the treated eyes. As such, it would appear that the rapid reduction in the level of hydrogen peroxide/sodium chlorite from the tears is not due to the loss of the tears of the subjects into the lachrymal duct. Rather, it is believed that the reduction is due to the presence of catalase and superoxide desmutase enzymes in the tears of human subjects. As the drops are placed in the eye of the patients, the catalase and other enzymes start the rapid enzymatic degradation of the hydrogen peroxide/sodium chlorite preparation, whereby in a matter of 3 minutes the level in the tears of the treated subjects is almost undetectable. The results of this experiment tend to show that upon instillation in the eye, the hydrogen peroxide/sodium chlorite mixture behaves like a self destructing preservative with the end products being water, oxygen, and sodium chloride.

(148) Additionally, the ocular symptoms of the treated eyes were observed and graded over a period of one hour post instillation for burning and stinging sensations, pain, redness, tearing, itching, photopsia, photophobia, discharge and for foreign body sensation as shown below.

(149) TABLE-US-00024 TABLE XIV Time post instillation Zero 30 1 2 3 5 10 15 30 60 time seconds minute minutes minutes minutes minutes minutes minutes minutes Burning/ 0 0 0 0 0 0 0 0 0 0 Stinging Pain 0 0 0 0 0 0 0 0 0 0 Redness 0 0 0 0 0 0 0 0 0 0 Tearing 0 0 0 0 0 0 0 0 0 0 Itching 0 0 0 0 0 0 0 0 0 0 Photopsia 0 0 0 0 0 0 0 0 0 0 Photophobia 0 0 0 0 0 0 0 0 0 0 Discharge 0 0 0 0 0 0 0 0 0 0 Foreign 0 +0.5 0 0 0 0 0 0 0 0 Body Sensation Grading Scale: 0 = None; +0.5 = Trace; +1 = Mild; +2 = Moderate; +3 = Moderately Severe; +4 = Severe

(150) The above results show that the hydrogen peroxide/sodium chlorite mixture is very well tolerated by the human eye without presenting any signs of irritation, inflammation, or any other adverse effects.

(151) It will be appreciated by those skilled in the art, that the invention has been described hereabove with reference to certain examples and specific embodiments. However, these are not the only examples and embodiments in which the invention may be practiced. Indeed, various modifications may be made to the above-described examples and embodiments without departing from the intended spirit and scope of the present invention. Accordingly, the present embodiments are to be considered on all respects as illustrative and not restrictive. It is intended that all such modifications be included within the scope of the following claims.