# US 10,765,732 B2 · Compounds and methods for affecting cytokines > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,765,732 B2 - Document ID: US10765732B2 - Status: Granted - Kind: B2 - Application number: 16/273,794 - Filed: 2019-02-12 - Published: 2019-06-13 - Granted: 2020-09-08 - Earliest claimed: 2013-06-13 - Assignees: ZIVO Bioscience, Inc. - Applicants: Zivo Bioscience, Inc. - Inventors: Andrew Dahl - Web page: https://patentgrep.com/patent/US10765732B2 ## CPC - A61K 39/092: {Streptococcus} - A61K 35/74: Bacteria (therapeutic use of a bacterial protein A61K38/00) - A61K 35/744: Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs - A61K 36/02: Algae - A61K 36/06: Fungi, e.g. yeasts - A61K 36/10: Bryophyta (mosses) - A61K 39/0258: {Escherichia} - A61K 39/09: {Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus}, streptococcus - 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) ## Abstract The present invention discloses isolates and fractions from a phyto-percolate and methods for affecting various cytokines by administering an effective amount of one or more of said isolates or fractions to an animal. In various exemplary embodiments, the isolates are useful for the treatment of bovine, canine and swine infection or inflammation, including bovine mastitis, by regulation of TNF-α, lactoferrin, IFN-γ, IL-1β, serum amyloid-A (SAA), IL-6 and/or β-defensin associated with infection or an immune response generally. ## Claims ### Claim 1 A method of treating mastitis in a cow, said method comprising administering to said cow a therapeutically effective amount of one or more liquid fractions of a sterile bioactive liquid obtained by filtering an incubated culture of ATCC Deposit No. PTA-5863 through sterilizing membrane filters and subjecting the sterile filtrate to chromatographic fractionation, wherein said one or more liquid fractions up-regulate at least one anti-inflammatory cytokine in bovine mammary epithelial cells, and/or down-regulate at least one pro-inflammatory cytokine in bovine mammary epithelial cells. ### Claim 2 (depends on 1) The method of claim 1, wherein each of said cytokines is selected from the group consisting of TNF-α, lactoferrin, IFN-γ, IL-1β, serum amyloid-A (SAA), IL-6 and β-defensin. ### Claim 3 (depends on 1) The method of claim 1, wherein said one or more liquid fractions each comprise at least one major bioactive species having a molecular weight of 452 Dalton or 678 Dalton. ### Claim 4 (depends on 1) The method of claim 1, wherein said one or more liquid fractions include at least one component identified by having a liquid chromatogram (LC) retention time of about 0.58 minutes, about 2.43 minutes, about 2.93 minutes, from about 3.14 to about 3.30 minutes, from about 3.50 to about 3.70 minutes, about 3.78 minutes, about 4.07 minutes, about 4.35 minutes, about 4.92 minutes, about 5.14 minutes, about 5.57 minutes, about 6.99 minutes, about 7.35 minutes, or about 7.63 minutes. ### Claim 5 A method of modulating an immune response in bovine, swine or canine, said method comprising the step of administering to said bovine, swine or canine an effective amount of one or more fractions of a sterile bioactive liquid obtained by filtering an incubated culture of ATCC Deposit No. PTA-5863 through sterilizing membrane filters and subjecting the sterile filtrate to chromatographic fractionation, wherein said immune response is characteristic of bovine mastitis, bovine respiratory disease complex, transition cow syndrome, canine osteoarthritis, canine skeletal-muscular overexertion, or porcine reproductive and respiratory syndrome virus immune disorder. ### Claim 6 (depends on 5) The method of claim 5, wherein said immune response is triggered by the presence of at least one of Staphylococcus aureus, Streptococcus uberis, and Escherichia coli. ### Claim 7 (depends on 5) The method of claim 5, wherein said one or more fractions of a sterile bioactive liquid up-regulate at least one anti-inflammatory cytokine, and/or down-regulate at least one pro-inflammatory cytokine involved in said immune response. ### Claim 8 (depends on 7) The method of claim 7, wherein each of said cytokines is selected from the group consisting of TNF-α, lactoferrin, IFN-γ, IL-1β, serum amyloid-A (SAA), IL-6 and β-defensin. ### Claim 9 (depends on 5) The method of claim 5, wherein said one or more fractions of a sterile bioactive liquid each comprise at least one major bioactive species having a molecular weight of 452 Dalton or 678 Dalton. ### Claim 10 (depends on 5) The method of claim 5, wherein said one or more fractions of a sterile bioactive liquid include at least one component identified by having a liquid chromatogram (LC) retention time of about 0.58 minutes, about 2.43 minutes, about 2.93 minutes, from about 3.14 to about 3.30 minutes, from about 3.50 to about 3.70 minutes, about 3.78 minutes, about 4.07 minutes, about 4.35 minutes, about 4.92 minutes, about 5.14 minutes, about 5.57 minutes, about 6.99 minutes, about 7.35 minutes, or about 7.63 minutes. ### Claim 11 A method of promoting an antimicrobial response at a site of infection in an animal, said method comprising the step of administering to said animal an effective amount of one or more liquid fractions of a sterile bioactive liquid obtained by filtering an incubated culture of ATCC Deposit No. PTA-5863 through sterilizing membrane filters and subjecting the sterile filtrate to chromatographic fractionation. ### Claim 12 (depends on 11) The method of claim 11, wherein said site of infection comprises the presence of Staphylococcus, Streptococcus, Escherichia, or Mycoplasma pathogens. ### Claim 13 (depends on 11) The method of claim 11, wherein said one or more liquid fractions of a sterile bioactive liquid each comprise at least one major bioactive species having a molecular weight of 452 Dalton or 678 Dalton. ### Claim 14 (depends on 11) The method of claim 11, wherein said one or more liquid fractions of a sterile bioactive liquid up-regulate at least one anti-inflammatory cytokine, and/or down-regulate at least one pro-inflammatory cytokine within cells at said site of infection. ### Claim 15 (depends on 14) The method of claim 14, wherein each of said cytokines is selected from the group consisting of TNF-α, lactoferrin, IFN-γ, IL-1β, serum amyloid-A (SAA), IL-6 and β-defensin. ### Claim 16 (depends on 14) The method of claim 14, wherein said animal is a cow, said cells comprise bovine mammary epithelial cells, and said site of infection is bovine mammary tissue.