# US 11,998,586 B2 · Compositions comprising antimicrobial peptides > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,998,586 B2 - Document ID: US11998586B2 - Status: Granted - Kind: B2 - Application number: 17/525,344 - Filed: 2021-11-12 - Published: 2022-02-24 - Granted: 2024-06-04 - Earliest claimed: 2019-11-08 - Assignees: University of Pittsburgh—Of the Commonwealth System of Higher Education - Applicants: University of Pittsburgh—Of the Commonwealth System of Higher Education - Inventors: Kenneth Urish; Jonathan Brendan Mandell - Web page: https://patentgrep.com/patent/US11998586B2 ## CPC - A61K 38/10: Peptides having 12 to 20 amino acids {(A61K38/043 - A61K38/046 take precedence)} - C07K 14/4723: {Cationic antimicrobial peptides, e.g. defensins} - 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 9/0024: {Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue (compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L)} - A61K 9/10: Dispersions; Emulsions - A61K 31/546: containing further heterocyclic rings, e.g. cephalothin - A61K 38/1729: {Cationic antimicrobial peptides, e.g. defensins} - A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like - A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics - A61P 31/04: Antibacterial agents - 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) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract A method of treating or preventing a microbial infection in a patient is provided, along with a wound irrigation system and a composition in the form of an irrigation liquid for reducing microbial load or preventing microbial infection in a wound. ## Claims ### Claim 1 A method of treating or preventing an infection in a subject in need thereof, comprising administering a pharmaceutical composition to an open wound, an implant, a medical device, a prosthetic, or any part thereof of the subject, wherein the infection comprises a biofilm, thereby treating or preventing the infection, wherein the pharmaceutical composition comprises: a. a cationic antimicrobial peptide or pharmaceutically acceptable salt thereof, wherein the cationic antimicrobial peptide is SEQ ID NO: 10; and b. a pharmaceutically acceptable aqueous carrier; wherein the pharmaceutical composition is in the form of a liquid; wherein the cationic antimicrobial peptide is present at a concentration of about 3 mg/mL to about 30 mg/mL, and wherein the pharmaceutical composition has a total osmolarity from about 50 mOsm/L to about 350 mOsm/L. ### Claim 2 (depends on 1) The method of claim 1, wherein the pharmaceutical composition further comprises a salt that is selected from the group consisting of sodium chloride, potassium chloride, potassium dihydrogen phosphate, disodium phosphate, calcium chloride, sodium lactate, copper chloride, copper sulfate, C11H22CuO14, ammonium hydroxide, magnesium hydroxide, sodium carbonate, and any combination thereof. ### Claim 3 (depends on 1) The method of claim 1, wherein the pharmaceutical composition further has a pH of from about 7.2 to about 8.0. ### Claim 4 (depends on 1) The method of claim 1, wherein the administering comprises irrigating the open wound, the implant, the medical device, the prosthetic, or any part thereof using the pharmaceutical composition. ### Claim 5 (depends on 4) The method of claim 4, wherein the irrigating further comprises reducing microbe load, wherein the microbe load is reduced by at least 1000-fold when the irrigating lasts for at most about 30 minutes. ### Claim 6 (depends on 1) The method of claim 1, wherein the treating or preventing the infection further comprises administering an antibiotic to the subject in need thereof. ### Claim 7 (depends on 1) The method of claim 1, wherein the aqueous carrier comprises water, normal saline, phosphate buffered saline, dextrose, glycerol, ethanol, buffered salt solutions, lactated Ringer's solution, or any combination thereof. ### Claim 8 (depends on 1) The method of claim 1, wherein the pharmaceutical composition further has a pH of from about 5.0 to about 8.0. ### Claim 9 (depends on 1) The method of claim 1, wherein the pharmaceutical composition further has a pH from about 7.4 to about 8.0. ### Claim 10 (depends on 1) The method of claim 1, wherein the infection comprises a bacteria species comprising Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species, Escherichia coli, or any combination thereof. ### Claim 11 (depends on 1) The method of claim 1, wherein the infection comprises periprosthetic joint infection. ### Claim 12 (depends on 1) The method of claim 1, wherein the method is subsequent to an irrigation and debridement (I&D) of the open wound, the implant, the medical device, the prosthetic, or any part thereof, wherein said I&D comprises use of a betadine, a chlorohexidine, or saline solution. ### Claim 13 (depends on 1) The method of claim 1, further comprising administering an antibiotic selected from the group consisting of: Aminoglycoside, Carbapenem, Ceftazidime, Cefepime, Ceftobiprole, Ceftaroline, Clindamycin, Cefazolin, Dalbavancin, Daptomycin, Fluoroquinolone, Linezolid, Mupirocin, Oritavancin, Piperacillin, Silver nitrate, Streptogramin, Telavancin, Tedizolid, Ticarcillin, Tigecycline, Vancomycin, and any combination thereof. ### Claim 14 (depends on 1) The method of claim 1, wherein the method reduces a bacterial burden of the biofilm by at least 102 CFU/mL as determined by a CFU quantification assay compared to an untreated subject. ### Claim 15 (depends on 1) The method of claim 1, wherein the method reduces a bacterial burden of the biofilm by at least a three-log reduction as determined by a CFU quantification assay compared to an untreated subject. ### Claim 16 (depends on 1) The method of claim 1, wherein the method reduces a bacterial burden of the biofilm by 99.9%. ### Claim 17 A method of treating or preventing an infection in a subject in need thereof, comprising irrigating an open wound, an implant, a medical device, a prosthetic, or any part thereof of the subject using a pharmaceutical composition, wherein the infection comprises a biofilm, thereby treating or preventing the infection, wherein the pharmaceutical composition comprises: a. a cationic antimicrobial peptide or pharmaceutically acceptable salt thereof, wherein the cationic antimicrobial peptide is SEQ ID NO: 10, and wherein the cationic antimicrobial peptide is present at a concentration of about 3 mg/mL to about 30 mg/mL; and b. a pharmaceutically acceptable aqueous carrier; wherein the pharmaceutical composition is in the form of a liquid; and wherein the irrigating is performed at least once a day. ### Claim 18 (depends on 17) The method of claim 17, wherein the cationic antimicrobial peptide is present at a concentration of about 10 mg/mL. ### Claim 19 (depends on 17) The method of claim 17, wherein the aqueous carrier comprises water, normal saline, phosphate buffered saline, dextrose, glycerol, ethanol, buffered salt solutions, lactated Ringer's solution, or any combination thereof. ### Claim 20 (depends on 17) The method of claim 17, wherein the pharmaceutical composition further has a pH of from about 5.0 to about 8.0. ### Claim 21 (depends on 17) The method of claim 17, wherein the pharmaceutical composition further has a pH from about 7.2 to about 8.0. ### Claim 22 (depends on 17) The method of claim 17, wherein the infection comprises a bacteria species comprising Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species, Escherichia coli, or any combination thereof. ### Claim 23 (depends on 17) The method of claim 17, wherein the infection comprises periprosthetic joint infection. ### Claim 24 (depends on 17) The method of claim 17, wherein the method is subsequent to an irrigation and debridement (I&D) of the open wound, the implant, the medical device, the prosthetic, or any part thereof, wherein said I&D comprises use of a betadine, a chlorohexidine, or saline solution. ### Claim 25 A method of treating or preventing an infection in a subject in need thereof, comprising irrigating an open wound, an implant, a medical device, a prosthetic, or any part thereof of the subject using a pharmaceutical composition, wherein the infection comprises a biofilm, thereby treating or preventing the infection, wherein the pharmaceutical composition comprises: a. a cationic antimicrobial peptide or pharmaceutically acceptable salt thereof, wherein the cationic antimicrobial peptide is SEQ ID NO: 10, and wherein the cationic antimicrobial peptide is present at a concentration of about 3 mg/mL to about 30 mg/mL; and b. a pharmaceutically acceptable aqueous carrier; wherein the pharmaceutical composition is in the form of a liquid; and wherein the irrigating using the pharmaceutical composition is performed for a period ranging from about 0.1 min to about 30 min. ### Claim 26 (depends on 25) The method of claim 25, wherein the irrigating using the pharmaceutical composition is performed for a period of about 15 min. ### Claim 27 (depends on 25) The method of claim 25, wherein the cationic antimicrobial peptide is present at a concentration of about 10 mg/mL. ### Claim 28 (depends on 25) The method of claim 25, wherein the aqueous carrier comprises water, normal saline, phosphate buffered saline, dextrose, glycerol, ethanol, buffered salt solutions, lactated Ringer's solution, or any combination thereof. ### Claim 29 (depends on 25) The method of claim 25, wherein the pharmaceutical composition further has a pH of from about 5.0 to about 8.0. ### Claim 30 (depends on 25) The method of claim 25, wherein the pharmaceutical composition further has a pH from about 7.2 to about 8.0. ### Claim 31 (depends on 25) The method of claim 25, wherein the infection comprises a bacteria species comprising Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species, Escherichia coli, or any combination thereof. ### Claim 32 (depends on 25) The method of claim 25, wherein the infection comprises periprosthetic joint infection. ### Claim 33 (depends on 25) The method of claim 25, wherein the method is subsequent to an irrigation and debridement (I&D) of the open wound, the implant, the medical device, the prosthetic, or any part thereof, wherein said I&D comprises use of a betadine, a chlorohexidine, or saline solution. ### Claim 34 A method of treating or preventing an infection in a subject in need thereof, comprising administering a pharmaceutical composition to an open wound, an implant, a medical device, a prosthetic, or any part thereof of the subject, wherein the infection comprises a biofilm, thereby treating or preventing the infection, wherein the pharmaceutical composition comprises: a. a cationic antimicrobial peptide or pharmaceutically acceptable salt thereof, wherein the cationic antimicrobial peptide is SEQ ID NO: 10, and wherein the cationic antimicrobial peptide is present at a concentration of about 3 mg/mL to about 30 mg/mL; and b. a pharmaceutically acceptable aqueous carrier; wherein the pharmaceutical composition is in the form of a liquid, and wherein the pharmaceutical composition is physiologically isotonic or physiologically hypotonic. ### Claim 35 (depends on 34) The method of claim 34, wherein the pharmaceutical composition is physiologically isotonic. ### Claim 36 (depends on 34) The method of claim 34, wherein the pharmaceutical composition is physiologically hypotonic. ### Claim 37 (depends on 34) The method of claim 34, wherein the aqueous carrier comprises water, normal saline, phosphate buffered saline, dextrose, glycerol, ethanol, buffered salt solutions, lactated Ringer's solution, or any combination thereof. ### Claim 38 (depends on 34) The method of claim 34, wherein the cationic antimicrobial peptide is present at a concentration of about 10 mg/mL. ### Claim 39 (depends on 34) The method of claim 34, wherein the pharmaceutical composition further has a pH of from about 5.0 to about 8.0. ### Claim 40 (depends on 34) The method of claim 34, wherein the pharmaceutical composition further has a pH from about 7.2 to about 8.0. ### Claim 41 (depends on 34) The method of claim 34, wherein the infection comprises a bacteria species comprising Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species, Escherichia coli, or any combination thereof. ### Claim 42 (depends on 34) The method of claim 34, wherein the infection comprises periprosthetic joint infection. ### Claim 43 (depends on 34) The method of claim 34, wherein the method is subsequent to an irrigation and debridement (I&D) of the open wound, the implant, the medical device, the prosthetic, or any part thereof, wherein said I&D comprises use of a betadine, a chlorohexidine, or saline solution. ### Claim 44 A method of treating or preventing an infection in a subject in need thereof, comprising administering a pharmaceutical composition to an open wound, an implant, a medical device, a prosthetic, or any part thereof of the subject, wherein the infection comprises a biofilm, thereby treating or preventing the infection, wherein the pharmaceutical composition comprises: a. a cationic antimicrobial peptide or pharmaceutically acceptable salt thereof, wherein the cationic antimicrobial peptide is SEQ ID NO: 10, and wherein the cationic antimicrobial peptide is present at a concentration of about 3 mg/mL to about 30 mg/mL; and b. a pharmaceutically acceptable aqueous carrier; wherein the pharmaceutical composition is in the form of a liquid; and wherein the pharmaceutical composition comprises a total ionic strength from about 0.02 M to about 0.2 M. ### Claim 45 (depends on 44) The method of claim 44, wherein the cationic antimicrobial peptide is present at a concentration of about 10 mg/mL. ### Claim 46 (depends on 44) The method of claim 44, wherein the aqueous carrier comprises water, normal saline, phosphate buffered saline, dextrose, glycerol, ethanol, buffered salt solutions, lactated Ringer's solution, or any combination thereof. ### Claim 47 (depends on 44) The method of claim 44, wherein the pharmaceutical composition further has a pH of from about 5.0 to about 8.0. ### Claim 48 (depends on 44) The method of claim 44, wherein the pharmaceutical composition further has a pH from about 7.2 to about 8.0. ### Claim 49 (depends on 44) The method of claim 44, wherein the infection comprises a bacteria species comprising Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species, Escherichia coli, or any combination thereof. ### Claim 50 (depends on 44) The method of claim 44, wherein the infection comprises periprosthetic joint infection. ### Claim 51 (depends on 44) The method of claim 44, wherein the method is subsequent to an irrigation and debridement (I&D) of the open wound, the implant, the medical device, the prosthetic, or any part thereof, wherein said I&D comprises use of a betadine, a chlorohexidine, or saline solution. ### Claim 52 A method of treating or preventing an infection in a subject in need thereof, comprising administering a pharmaceutical composition to an open wound, an implant, a medical device, a prosthetic, or any part thereof of the subject, wherein the infection comprises a biofilm, thereby treating or preventing the infection, wherein the pharmaceutical composition comprises: a. a cationic antimicrobial peptide or pharmaceutically acceptable salt thereof, wherein the cationic antimicrobial peptide is SEQ ID NO: 10, and wherein the cationic antimicrobial peptide is present at a concentration of about 10 mg/mL; and b. a pharmaceutically acceptable aqueous carrier; wherein the pharmaceutical composition is in the form of a liquid. ### Claim 53 (depends on 52) The method of claim 52, wherein the aqueous carrier comprises water, normal saline, phosphate buffered saline, dextrose, glycerol, ethanol, buffered salt solutions, lactated Ringer's solution, or any combination thereof. ### Claim 54 (depends on 52) The method of claim 52, wherein the pharmaceutical composition further has a pH of from about 5.0 to about 8.0. ### Claim 55 (depends on 52) The method of claim 52, wherein the pharmaceutical composition further has a pH from about 7.2 to about 8.0. ### Claim 56 (depends on 52) The method of claim 52, wherein the infection comprises a bacteria species comprising Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species, Escherichia coli, or any combination thereof. ### Claim 57 (depends on 52) The method of claim 52, wherein the infection comprises periprosthetic joint infection. ### Claim 58 (depends on 52) The method of claim 52, wherein the method is subsequent to an irrigation and debridement (I&D) of the open wound, the implant, the medical device, the prosthetic, or any part thereof, wherein said I&D comprises use of a betadine, a chlorohexidine, or saline solution. ### Claim 59 A method of treating or preventing an infection in a subject in need thereof, comprising injecting a pharmaceutical composition to an open wound, an implant, a medical device, a prosthetic, or any part thereof of the subject, thereby treating or preventing the infection, wherein the pharmaceutical composition comprises: a. a cationic antimicrobial peptide or pharmaceutically acceptable salt thereof, wherein the cationic antimicrobial peptide is SEQ ID NO: 10, and wherein the cationic antimicrobial peptide is present at a concentration of about 3 mg/mL to about 30 mg/mL; and b. a pharmaceutically acceptable aqueous carrier; wherein the pharmaceutical composition is in the form of a liquid. ### Claim 60 (depends on 59) The method of claim 59, wherein the cationic antimicrobial peptide is present at a concentration of about 10 mg/mL. ### Claim 61 (depends on 59) The method of claim 59, wherein the aqueous carrier comprises water, normal saline, phosphate buffered saline, dextrose, glycerol, ethanol, buffered salt solutions, lactated Ringer's solution, or any combination thereof. ### Claim 62 (depends on 59) The method of claim 59, wherein the pharmaceutical composition further has a pH of from about 5.0 to about 8.0. ### Claim 63 (depends on 59) The method of claim 59, wherein the pharmaceutical composition further has a pH from about 7.2 to about 8.0. ### Claim 64 (depends on 59) The method of claim 59, wherein the infection comprises a bacteria species comprising Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species, Escherichia coli, or any combination thereof. ### Claim 65 (depends on 59) The method of claim 59, wherein the infection comprises periprosthetic joint infection. ### Claim 66 (depends on 59) The method of claim 59, wherein the method is subsequent to an irrigation and debridement (I&D) of the open wound, the implant, the medical device, the prosthetic, or any part thereof, wherein said I&D comprises use of a betadine, a chlorohexidine, or saline solution.