# US 8,784,898 B2 · Methods of wound care and treatment

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- Number: US 8,784,898 B2
- Document ID: US8784898B2
- Status: Granted
- Kind: B2
- Application number: 12/769,323
- Filed: 2010-04-28
- Published: 2011-04-07
- Granted: 2014-07-22
- Earliest claimed: 2006-10-25
- Assignees: Revalesio Corporation
- Applicants: Gregory J. Archambeau; Richard L. Watson; Anthony B. Wood
- Inventors: Gregory J. Archambeau; Richard L. Watson; Anthony B. Wood
- Web page: https://patentgrep.com/patent/US8784898B2

## CPC

- A61K 33/00: Medicinal preparations containing inorganic active ingredients
- A61K 9/0014: {Skin, i.e. galenical aspects of topical compositions (non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00)}
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 9/0048: {Eye, e.g. artificial tears}
- A61K 9/06: Ointments; Bases therefor; {Other semi-solid forms, e.g. creams, sticks, gels (composition of ointments, creams or gels A61K47/00)}
- A61K 9/08: Solutions {(composition of solutions A61K47/00)}
- A61K 9/14: Particulate form, e.g. powders, {Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles (microspheres A61K9/16; microcapsules A61K9/50; nanocapsules, nanoparticles of the matrix type A61K9/51)}
- A61K 9/5192: {Processes}
- A61K 9/7007: {Drug-containing films, membranes or sheets (A61K9/0041, A61K9/0043, A61K9/006, A61K9/0063 take precedence)}
- A61K 31/573: substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
- A61K 31/58: containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin (digitoxin {A61K31/7048})
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61K 47/02: Inorganic compounds
- A61K 47/36: Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K 47/42: Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein (oligopeptides having up to five amino acids {A61K47/183}; polyamino acids A61K47/34)
- A61L 15/44: Medicaments
- A61L 15/46: Deodorants or malodour counteractants, e.g. to inhibit the formation of ammonia or bacteria
- A61L 26/0066: {Medicaments; Biocides}
- A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/04: Antibacterial agents
- B01F 23/233: using driven stirrers with completely immersed stirring elements
- B01F 23/41: Emulsifying
- B01F 23/43: using driven stirrers
- B01F 27/2722: {provided with ribs, ridges or grooves on one surface}
- B29D 11/00038: {Production of contact lenses}
- A61L 2300/10: containing or releasing inorganic materials
- A61L 2300/404: Biocides, antimicrobial agents, antiseptic agents
- A61L 2300/41: Anti-inflammatory agents, e.g. NSAIDs
- A61L 2300/412: Tissue-regenerating or healing or proliferative agents
- A61L 2300/436: of receptors
- A61L 2400/12: Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- B01F: MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING (mixing paints B44D3/06)
- B29D: PRODUCING PARTICULAR ARTICLES FROM PLASTICS OR FROM SUBSTANCES IN A PLASTIC STATE (making granules B29B9/00; making preforms B29B11/00)

## Abstract

Provided are electrokinetically-altered fluids (e.g., gas-enriched electrokinetic fluids) comprising an ionic aqueous solution of charge-stabilized oxygen-containing nanostructures in an amount sufficient to provide modulation of at least one of cellular membrane potential and cellular membrane conductivity, and therapeutic compositions and methods for use in treating a wound to a surface tissue or a symptom thereof. The electrokinetically-altered fluids or therapeutic compositions and methods include electrokinetically-altered ionic aqueous fluids optionally in combination with other therapeutic agents. Particular aspects provide for regulating or modulating intracellular signal transduction associated with said inflammatory responses by modulation of at least one of cellular membranes, membrane potential, membrane proteins such as membrane receptors, including but not limited to G-Protein Coupled Receptors (GPCR), and intercellular junctions (e.g., tight junctions, gap junctions, zona adherins and desmasomes). Other embodiments include particular routes of administration or formulations for the electrokinetically-altered fluids (e.g., electrokinetically-altered gas-enriched fluids and solutions) and therapeutic compositions.

## Claims

### Claim 1

A method for treating a wound to a surface tissue or a symptom thereof, comprising administering to a subject in need thereof a therapeutically effective amount of an electrokinetically-altered aqueous fluid comprising an ionic aqueous solution of charge-stabilized oxygen-containing nanobubbles having an average diameter of less than 80 nanometers and stably configured in the ionic aqueous fluid in an amount sufficient for treating a wound to a surface tissue or a symptom thereof.

### Claim 2 (depends on 1)

The method of claim 1, wherein treating the wound comprises treating to decrease scarring in the wound to the surface tissue, wherein scarring is reduced.

### Claim 3 (depends on 2)

The method of claim 2, wherein the electrokinetically-altered aqueous fluid further increases the level of chemokines at the wound.

### Claim 4 (depends on 1)

The method of claim 1, wherein treating the wound comprises treating to increase or decrease nitric oxide production or degradation in the wound to the surface tissue, wherein an increased or decreased level of nitric oxide is afforded.

### Claim 5 (depends on 1)

The method of claim 1, wherein the electrokinetically-altered aqueous fluid is contacted to the wound by means of a wound dressing.

### Claim 6 (depends on 1)

The method of claim 1, wherein the electrokinetically-altered aqueous fluid accelerates epidermal or dermal layering.

### Claim 7 (depends on 1)

The method of claim 1, wherein the electrokinetically-altered aqueous fluid increases cellular migration of at least one type of cell to the wound.

### Claim 8 (depends on 7)

The method of claim 7, wherein the type of cellular migration or proliferation comprises at least one cell selected from the group consisting of: keratinocytes, fibroblasts, epidermal cells, dermal cells, epithelial cells, mast cells, neutrophils, lymphocytes, and macrophages.

### Claim 9 (depends on 1)

The method of claim 1, wherein the electrokinetically-altered aqueous fluid accelerates neoangiogenesis of blood vessels or lymphatic vessels.

### Claim 10 (depends on 1)

The method of claim 1, wherein the electrokinetically-altered aqueous fluid increases collagen deposition at the wound.

### Claim 11 (depends on 1)

The method of claim 1, wherein the electrokinetically-altered aqueous fluid reduces growth of at least one microbe in the wound.

### Claim 12 (depends on 11)

The method of claim 11, wherein the microbe comprises Pseudomonas.

### Claim 13 (depends on 1)

The method of claim 1, wherein the charge-stabilized oxygen-containing nanobubbles are the major charge-stabilized gas-containing nanostructure species in the fluid.

### Claim 14 (depends on 1)

The method of claim 1, wherein the percentage of dissolved oxygen molecules present in the fluid as the charge-stabilized oxygen-containing nanobubbles is greater than 0.01%.

### Claim 15 (depends on 1)

The method of claim 1, wherein the percentage of dissolved oxygen present in the fluid as the charge-stabilized oxygen-containing nanobubbles is a percentage greater than 50%.

### Claim 16 (depends on 1)

The method of claim 1, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 70 nm.

### Claim 17 (depends on 1)

The method of claim 1, wherein the ionic aqueous solution comprises a saline solution.

### Claim 18 (depends on 1)

The method of claim 1, wherein the fluid is superoxygenated.

### Claim 19 (depends on 1)

The method of claim 1, wherein the fluid comprises a form of solvated electrons.

### Claim 20 (depends on 1)

The method of claim 1, wherein alteration of the electrokinetically altered aqueous fluid comprises exposure of the fluid to hydrodynamically-induced, localized electrokinetic effects.

### Claim 21 (depends on 20)

The method of claim 20, wherein exposure to the localized electrokinetic effects comprises exposure to at least one of voltage pulses and current pulses.

### Claim 22 (depends on 20)

The method of claim 20, wherein the exposure of the fluid to hydrodynamically-induced, localized electrokinetic effects comprises exposure of the fluid to electrokinetic effect-inducing structural features of a device used to generate the fluid.

### Claim 23 (depends on 1)

The method of claim 1, wherein the wound to a surface tissue or the symptom thereof comprises at least one wound type selected from the group consisting of lacerations, abrasions, rupture, puncture wounds, chemical, thermal, or radiation-induced burns, cuts, scrapes, incisions, blisters, diabetic ulcers, bedsores or pressure ulcers, skin grafts, and surgical wounds.

### Claim 24 (depends on 23)

The method of claim 23, wherein the wound to a surface tissue or the symptom thereof comprises at least one of diabetic ulcers, bedsores or pressure ulcers, skin grafts, and surgical wounds.

### Claim 25 (depends on 24)

The method of claim 24, wherein the wound to a surface tissue or the symptom thereof comprises diabetic ulcers and bedsores or pressure ulcers.

### Claim 26 (depends on 1)

The method of claim 1, wherein the at least one symptom comprising at least one condition selected from the group consisting of epidermal or dermal layering, cellular migration, collagen deposition at the wound, neoangiogenesis at the wound, elastin deposition at the wound, inflammation at the wound, expression of proteoglycans or glycosaminoglycans, cellular proliferation, and hyaluronic acid concentration at the wound.

### Claim 27 (depends on 1)

The method of claim 1, wherein the electrokinetically altered aqueous fluid modulates localized or cellular levels of nitric oxide.

### Claim 28 (depends on 1)

The method of claim 1 wherein the electrokinetically altered aqueous fluid promotes a localized decrease at the site of administration of at least one cytokine selected from the group consisting of: IL-1beta, IL-8, TNF-alpha, and TNF-beta.

### Claim 29 (depends on 1)

The method of claim 1, further comprising a synergistic or non-synergistic inhibition or reduction in inflammation by simultaneously or adjunctively treating the subject with another anti-inflammatory agent.

### Claim 30 (depends on 29)

The method of claim 29, wherein said other anti-inflammatory agent comprises a steroid or glucocorticoid steroid.

### Claim 31 (depends on 30)

The method of claim 30, wherein said steroid comprises a glucocorticoid steroid.

### Claim 32 (depends on 1)

The method of claim 1, further comprising combination therapy, wherein at least one additional therapeutic agent is administered to the patient.

### Claim 33 (depends on 32)

The method of claim 32, wherein the one additional therapeutic agent is selected from the group consisting of anti-microbial agents, antifungal agents, and antibiotic agents.

### Claim 34 (depends on 32)

The method of claim 32, wherein, the at least one additional therapeutic agent is selected from the group consisting of: ciclosporin, hyaluronic acid, carmellose, macrogol(s), dextran and hyprolose, sodium and calcium, sodium and povidone, hypromellose, carbomer, amikacin, gentamicin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin, paromomycin, geldanamycin, herimycin, loracarbef, ertapenem, imipenem/cilastatin, meropenem, cefadroxil, cefazolin, cefalotin/cefalothin, cephalexin, cefaclor, cefamandole, cefoxitin, cefuroxime, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefeprime, teicoplanin, vancomycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spectinomycin, aztreonam, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, nafcillin, penicillin, peperacillin, ticarcillin, bacitracin, colistin, polymyxin B, ciprofloxacin, enoxacin, gatifloxacin, levofloxacin, lomefloxacin, moxifloxacin, norfloxacin, ofloxacin, trovafloxacin, mafenide, protosil, sulfacetamide, sulfamethizole, sulfanilamide, sulfasalazine, sulfisoxazole, trimethoprim, trimethoprim-sulfamethoxazole, demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline, arsphenamine, chloramphenicol, clindamycin, lincoamycin, ethambutol, fosfomycin, fusidic acid, furazolidone, isoniazid, linezolid, metronidazole, mupirocin, nitrofurantoin, platensimycin, pyrazinamide, quinupristin/dalfopristin, rifampin/rifampicin, tinidazole, miconazole, ketoconazole, clotrimazole, econazole, bifonazole, butoconazole, fenticonazole, isoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, fluconazole, itraconazole, isavuconazole, ravuconazole, posaconazole, voriconazole, teronazole, terbinafine, amorolfine, naftifine, butenafine, anidulafungin, caspofungin, micafungin, ciclopirox, flucytosine, griseofulvin, Gentian violet, haloprogin, tolnaftate, undecylenic acid, and combinations thereof.

### Claim 35 (depends on 32)

The method of claim 32, wherein the at least one additional therapeutic agent is a thymic stromal lymphopoictin (TSLP) and/or thymic stromal lymphopoietin receptor (TSLPR) antagonist.

### Claim 36 (depends on 35)

The method of claim 35, wherein the TSLP and/or TSLPR antagonist is selected from the group consisting of neutralizing antibodies specific for TSLP and the TSLP receptor, soluble TSLP receptor molecules, and TSLP receptor fusion proteins, including TSLPR-immunoglobulin Fc molecules or polypeptides that encode components of more than one receptor chain.

### Claim 37 (depends on 1)

The method of claim 1, wherein the charge-stabilized oxygen-containing nanobubbles are stably configured in the ionic aqueous fluid in an amount sufficient to provide, upon contact of a living cell by the fluid, modulation of at least one of cellular membrane potential and cellular membrane conductivity.

### Claim 38 (depends on 37)

The method of claim 37, wherein modulation of at least one of cellular membrane potential and cellular membrane conductivity comprises altering of a conformation, ligand binding activity, or a catalytic activity of a membrane associated protein.

### Claim 39 (depends on 38)

The method of claim 38, wherein the membrane associated protein comprises at least one selected from the group consisting of receptors, transmembrane receptors, ion channel proteins, intracellular attachment proteins, cellular adhesion proteins, integrins, etc.

### Claim 40 (depends on 39)

The method of claim 39, wherein the transmembrane receptor comprises a G-Protein coupled receptor (GPCR).

### Claim 41 (depends on 40)

The method of claim 40, wherein the GPCR interacts with a G protein α subunit.

### Claim 42 (depends on 41)

The method of claim 41, wherein the G protein α subunit comprises at least one selected from the group consisting of Gαs, Gαi, Gαq, and Gα12.

### Claim 43 (depends on 41)

The method of claim 41, wherein the at least one G protein α subunit is Gαq.

### Claim 44 (depends on 1)

The method of claim 1, wherein modulating cellular membrane conductivity comprises modulating whole-cell conductance.

### Claim 45 (depends on 44)

The method of claim 44, wherein modulating whole-cell conductance, comprises modulating at least one of linear and non-linear voltage-dependent contribution of the whole-cell conductance.

### Claim 46 (depends on 37)

The method of claim 37, wherein modulation of at least one of cellular membrane potential and cellular membrane conductivity comprises modulation of a calcium dependent cellular messaging pathway or system.

### Claim 47 (depends on 37)

The method of claim 37, wherein modulation of at least one of cellular membrane potential and cellular membrane conductivity comprises modulation of phospholipase C activity.

### Claim 48 (depends on 37)

The method of claim 37, wherein modulation of at least one of cellular membrane potential and cellular membrane conductivity comprises modulation of adenylate cyclase (AC) activity.

### Claim 49 (depends on 37)

The method of claim 37, wherein modulation of at least one of cellular membrane potential and cellular membrane conductivity comprises modulation of intracellular signal transduction associated with at least one condition or symptom selected from the group consisting of epidermal or dermal layering, cellular migration, collagen deposition at the wound, neoangiogenesis at the wound, elastin deposition at the wound, inflammation at the wound, expression of proteoglycans or glycosaminoglycans, cellular proliferation, and hyaluronic acid concentration at the wound.

### Claim 50 (depends on 1)

The method of claim 1, comprising administration to a cell network or layer, and further comprising modulation of an intercellular junction therein.

### Claim 51 (depends on 50)

The method of claim 50, wherein the intracellular junction comprises at least one selected from the group consisting of tight junctions, gap junctions, zona adherins and desmasomes.

### Claim 52 (depends on 50)

The method of claim 50, wherein the cell network or layer comprises at least one selected from the group consisting of epithelial cell layer or network.

### Claim 53 (depends on 1)

The method of claim 1, wherein the electrokinetically-altered aqueous fluid is oxygenated, and wherein the oxygen in the fluid is present in an amount of at least 8 ppm at atmospheric pressure.

### Claim 54 (depends on 1)

The method of claim 1, wherein the electrokinetically altered aqueous fluid comprises at least one of a form of solvated electrons and electrokinetically modified or charged oxygen species.

### Claim 55 (depends on 54)

The method of claim 54, wherein the form of solvated electrons or electrokinetically modified or charged oxygen species are present in an amount of at least 0.01 ppm.

### Claim 56 (depends on 54)

The method of claim 54, wherein the electrokinetically-altered aqueous fluid comprises a form of solvated electrons stabilized, at least in part, by molecular oxygen.

### Claim 57 (depends on 37)

The method of claim 37, wherein the ability to modulate of at least one of cellular membrane potential and cellular membrane conductivity persists for at least two months in a closed gas-tight container.

### Claim 58 (depends on 38)

The method of claim 38, wherein the membrane associated protein comprises CCR3.

### Claim 59 (depends on 1)

The method of claim 1, wherein treating a wound to a surface tissue or a symptom thereof, comprises modulation of intracellular NF-κB expression and/or activity.

### Claim 60 (depends on 1)

The method of claim 1, wherein treating comprises topical administration to the wound.

### Claim 61 (depends on 1)

The method of claim 1, wherein the electrokinetically-altered fluid comprises at least one cation selected from the group consisting of ammonium, calcium, copper, iron, magnesium, manganese, potassium, sodium and zinc.

### Claim 62 (depends on 1)

The method of claim 1, wherein a majority of the charge-stabilized oxygen-containing nanobubbles have a diameter of less than 80 nanometers.

### Claim 63 (depends on 16)

The method of claim 16, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 60 nm.

### Claim 64 (depends on 63)

The method of claim 63, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 50 nm.

### Claim 65 (depends on 64)

The method of claim 64, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 40 nm.

### Claim 66 (depends on 65)

The method of claim 65, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 30 nm.

### Claim 67 (depends on 66)

The method of claim 66, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 20 nm.

### Claim 68 (depends on 67)

The method of claim 67, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 10 nm.

### Claim 69 (depends on 68)

The method of claim 68, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 5 nm.

### Claim 70 (depends on 69)

The method of claim 69, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 4 nm.

### Claim 71 (depends on 70)

The method of claim 70, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 3 nm.

### Claim 72 (depends on 71)

The method of claim 71, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 2 nm.

### Claim 73 (depends on 72)

The method of claim 72, wherein the charge-stabilized oxygen-containing nanobubbles have an average diameter of less than 1 nm.