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WO2018191409A1 - Agents antimicrobiens et leurs procédés d'utilisation - Google Patents

Agents antimicrobiens et leurs procédés d'utilisation Download PDF

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Publication number
WO2018191409A1
WO2018191409A1 PCT/US2018/027157 US2018027157W WO2018191409A1 WO 2018191409 A1 WO2018191409 A1 WO 2018191409A1 US 2018027157 W US2018027157 W US 2018027157W WO 2018191409 A1 WO2018191409 A1 WO 2018191409A1
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WIPO (PCT)
Prior art keywords
pca
metabolite
composition
skin
anthocyanin
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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PCT/US2018/027157
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English (en)
Inventor
Lanny Leo Johnson
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Individual
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Individual
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Publication date
Priority claimed from US15/484,827 external-priority patent/US10016380B2/en
Application filed by Individual filed Critical Individual
Publication of WO2018191409A1 publication Critical patent/WO2018191409A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/11—Aldehydes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/42—Use of materials characterised by their function or physical properties
    • A61L15/46—Deodorants or malodour counteractants, e.g. to inhibit the formation of ammonia or bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/216—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with other specific functional groups, e.g. aldehydes, ketones, phenols, quaternary phosphonium groups
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention generally relates to antimicrobial compositions, methods for promoting healing of a wound and more specifically to methods and compositions including the administration of an anthocyanin or an anthocyanidin or metabolites thereof for promoting wound healing by reducing or preventing microbial growth and inducing the activation of growth hormones.
  • nutraceutical compositions containing anthocyanin or anthocyanidins or metabolites thereof suitable for administration to a mammal for promoting or inducing wound healing are also described.
  • compositions and methods of sanitizing skin antiseptic or sterilizing solid surfaces, porous or semi-porous or cloth like surfaces, such as materials or cloth or bandages.
  • compositions and methods for killing bacterial biofilms which may be present on implantable devices, solid surfaces, porous or semi-porous, or cloth-like material.
  • the Centers for Disease Control and Prevention estimated at least 2 million people in the United States become infected with bacteria that are resistant to antibiotics and at least 23,000 people die each year as a direct result of these infections. (Antibiotic Resistance Threats in the United States, 2013, Centers for Disease Control and Prevention, Atlanta, Ga., USA 2013).
  • the CDC report classified three microorganisms with an antibiotic resistance threat level of urgent in the United States and twelve microorganisms with an antibiotic resistance threat level of serious.
  • Clostridium difficile Carbapenem-resistant Enterobacteriacaeae (CRE) and drug resistant Neisseria gonorrhoeae (cephalosporin resistance) are classified by the CDC as urgent because they require urgent public health attention to identify infections and to limit transmission.
  • CRE Carbapenem-resistant Enterobacteriacaeae
  • Neisseria gonorrhoeae cephalosporin resistance
  • Clostridium difficile is the most frequent etiologic agent for health-care-associated diarrhea. In one hospital, 30% of adults who developed health-care-associated diarrhea were positive for C. difficile.
  • Risk factors for acquiring C. difficile-associated infection include a) exposure to antibiotic therapy, particularly with beta-lactam agents; b) gastrointestinal procedures and surgery; c) advanced age; and d) indiscriminate use of antibiotics.
  • C. difficile is an anaerobic, gram-positive bacterium capable of sporulating when environmental conditions no longer support its continued growth.
  • the capacity to form spores enables the organism to persist in the environment (e.g., on dry surfaces or in soil) for extended periods of time.
  • Environmental contamination by this microorganism is well known, especially direct exposure to contaminated patient-care items and high-touch surfaces in patients' bathrooms have been implicated as sources of infection.
  • EBLs Enterobacteriacaea
  • VRE Vancomycin-resistant Enterococcus
  • Multidrug- resistant Psuedomonas Aeruginosa Drug-resistant Non-typhoidal Salmonella, Drug- resistant Salmonella Typhi, Drug-resistant Shigella, Methicillin-resistant Staphylococcus auereus (MRSA), Drug-resistant Streptococcus pneumonia, Drug-resistant tuberculosis (MDR and XDR)(Antibiotic Resistance Threats in the United States, 2013, Centers for Disease Control and Prevention, Atlanta, Ga., USA 2013).
  • MRSA Methicillin-resistant Staphylococcus auereus
  • MDR and XDR Drug-resistant Streptococcus pneumonia
  • Staphylococcus aureus is the most frequently identified antimicrobial drug- resistant pathogen in U.S. hospitals. MRSA was one of the first pathogens to develop resistance, first detected the United Kingdom in 1961. In 1999, MRSA was responsible for 37% of fatal cases of sepsis in the UK. Additionally, half of all S. aureus infections in the U.S. are resistant to penicillin, methicillin, tetracycline and erythromycin, leaving only vancomycin as an effective agent against S.
  • GISA glycopeptide-intermediate Staphylococcus aureus
  • VSA vancomycin-intermediate Staphylococcus aureus
  • CA-MRSA community-acquired MRSA
  • Treatment of the failed infected total joint may include repeat surgery, removal of the implant, insertion of an antibiotic impregnated spacer, hospitalization, therapy and return at later date to remove spacer and redo the total joint with
  • Candida albicans is the major fungal pathogen of humans. (Warren, NG, American Society for Microbiology; 1995. 723; Bachmann, SP, Antimicrobial Agents Chemother, 2002; 46: 3591 ). It has been reported that mortality rate of patients with catheter related candidemia approaches 40%. (Fux, CA, Trends Microbiol, 2005; 13(1 ): 34; and Tampakakis, E., Eukaryot Cell, 2009; 8:732). Biofilms of C. albicans are capable of holding other micro-organisms and more likely to be heterogeneous with other bacteria and fungi in the environment and on medical devices.
  • biofilm cells are significantly less susceptible to antimicrobial agents.
  • drug therapy for an implant infection may be futile, and often, the only solution is mechanical removal of the implant.
  • Biofilm formation also plays an important role in outbreaks of C. albicans related infections.
  • ChloraPrep (2% chlorhexidine gluconate in 70% isopropyl alcohol) for skin antiseptic in preparation for blood culture collection. Journal of Infection , Volume 57, Issue 6 , 459 - 463). This is likely due to the fact that P. acnes normally reside deep in the skin surface within the hair follicles and/or sebaceous glands. Therefore, it necessary to have a composition of matter in treating potential pathogens on the human skin, especially P. acnes that contains within the vehicle a property than enhances skin penetration.
  • Nakatsuji et al showed that the microbiota extends within the dermis, therefore, enabling physical contact between bacteria and various cells below the basement membrane. These observations show that normal commensal bacterial communities directly communicate with the host in a tissue previously thought to be sterile. (Nakatsuji T, Chiang HI, Jiang SB, Nagarajan H, Zengler K, Gallo RL. The microbiome extends to subepidermal compartments of normal skin. Nat Commun. 2013;4: 1431 .) Zeeuwen et. al. showed the bacterial communities of the surface of human skin, mostly under static conditions in healthy volunteers differs from what is found following skin injury. The dynamics of re-colonization of skin microbiota following skin barrier disruption by tape stripping as a model of superficial injury showed microbiome of the deeper layers, rather than that of the superficial skin layer, may be regarded as the host indigenous
  • biofilms are highly resistant to most antimicrobial agents and disinfectants (Donlan RM. Role of biofilms in antimicrobial resistance. ASAIO J. 2000;46:S47-52.).
  • Biofilms may form on a wide variety of surfaces, including living tissues, indwelling medical devices, industrial or potable water system piping, or natural aquatic systems.
  • An implant in the body i.e. total joint, typically has the biofilms attach to the implant and then the bacteria colony grows.
  • a common example of biofilms is dental plaque. It requires physical removal with a tooth brush.
  • Donlan reported that biofilms are highly resistant to most antimicrobial agents and disinfectants. (Donlan RM. Role of biofilms in antimicrobial resistance.
  • MRSA Staphylococcus aureus
  • the present invention provides generally for compositions and methods for reducing or preventing microbial growth, which are particularly useful for treating and promoting the healing of wounds. Specifically, the present invention provides methods and pharmaceutical and nutraceutical compositions that reduce or substantially eliminate potentially pathologic microbes.
  • a method of promoting healing of a wound in a mammal comprising administering an anthocyanin or an anthocyanidin to the mammal in need of such treatment a therapeutically effective amount of the anthocyanin or anthocyanidin composition or compound wherein microbial growth is reduced and local growth hormone activity is optimized.
  • a method of promoting healing of a wound in a mammal comprising administering an anthocyanin metabolite or an anthocyanidin metabolite to the mammal in need of such treatment a therapeutically effective amount of the metabolite or anthocyanidin metabolite compositions wherein microbial growth is reduced and local growth hormone activity is optimized.
  • the invention provides methods and compositions that effectively deliver the compositions to the affected wound.
  • the present invention provides compositions and methods for use in the treatment of a variety of wound problems, including but not limited to, burns, pressure wounds, abrasions, diabetic wounds, peri-surgical and skin infections.
  • the present invention provides compositions and methods for use in the treatment of a variety of infections, including those from blood transfusions.
  • the therapeutic compositions and compounds are administered orally or topically; however, the therapeutic compositions and compounds may be administered by any conventional route including, for example, oral, topical, buccal, injection, pulmonary, intravenous, inhalant, subcutaneous, sublingual, or transdermal.
  • Those of skill in the art can readily determine the various parameters and conditions for producing these compositions or formulations without resort to undue experimentation.
  • the present invention provides a pharmaceutical composition for promoting wound healing, comprising: a) an anthocyanin; or b) anthocyanidin.
  • the anthocyanin can be selected from cyanidin-3- glucoside or delphinidin-3-glucoside, cyanidin-3-galactoside, and pelargonidin-3- galactoside.
  • the anthocyanidins can be selected from cyanidin, delphinidin, pelargonidin, malvidin and petunidin.
  • the present disclosure provides a
  • composition comprising: a) an anthocyanin metabolite; or b) an anthocyanidin metabolite.
  • a pharmaceutical composition for promoting wound healing comprising: a) an anthocyanin metabolite; or b) an anthocyanidin metabolite.
  • metabolites can be selected from protocatechuic acid and 2, 3, 4
  • the present disclosure provides a pharmaceutical composition comprising: a) a metabolite of an anthocyanin metabolite; or b) a metabolite of an anthocyanidin metabolite.
  • the metabolites are metabolites selected from protocatechuic acid, and 2, 3, 4 trihydroxybenzaldehyde.
  • the pharmaceutical composition can comprise: a) an anthocyanin; or b) an anthocyanidin; and c) a pharmaceutically acceptable carrier.
  • the pharmaceutical composition can comprise: a) an anthocyanin metabolite; or b) an anthocyanidin metabolite; and c) a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier can be selected from, but not limited to, any carrier, diluent or excipient compatible with the other ingredients of the composition.
  • the pharmaceutical composition can comprise: a) an anthocyanin; and/or b) an anthocyanidin; and/or c) a pharmaceutically acceptable carrier; and/or d) an acceptable delivery carrier.
  • the delivery carrier can be formulated and administered as known in the art, e.g., for topical, oral, buccal, injection, intravenous, inhalant, subcutaneous, sublingual or transdermal
  • said topical delivery carrier may be formulated and administered to any surface, including but not limited to skin, bone, synovium, cartilage, and implants.
  • the acceptable delivery carrier can be selected from any dermal or
  • the acceptable delivery carrier is a biodegradable microsphere or a slow release bioadsorbable material.
  • the acceptable delivery carrier can be selected from 50/50 D, L lactide/glycolide or 85/15 D, L lactide/glycolide, both of which are amorphous physically and, therefore, are non-reactive when used as a carrier in a composition that is delivered in or to the body.
  • the anthocyanidin provided in any recited composition is provided less than 200 mM and in other embodiments about 100 mM. In some embodiments, the anthocyanin or anthocyanidin provided in any recited composition or method of use is provided in a range of between 10 to 200 mM. In other embodiments, the anthocyanin metabolite or anthocyanidin metabolite provided in any recited composition or method of use is provided in a range of between 20 to 200 mM.
  • the anthocyanin, anthocyanidin, or metabolites thereof provided in any recited composition or method of use is provided in a range of between 20 to 100 mM. In yet other embodiments, the anthocyanin, anthocyanidin, or metabolites thereof provided in any recited composition or method of use is provided in a range of between 20 to 50 mM. In some embodiments, the metabolite is provided in any recited composition less than 100 mM and in others at 25 mM.
  • an antimicrobial composition comprising: a) an anthocyanin metabolite; or b) an
  • anthocyanidin metabolite can be selected from protocatechuic acid, 2, 3, 4 trihydroxybenzaldehyde.
  • metabolites can be selected from protocatechuic acid, 2, 3, 4 trihydroxybenzaldehyde.
  • anthocyanin, anthocyanidin and metabolites thereof can have broad spectrum activity against disease-causing microbes.
  • a method of treating a wound of an individual comprising administering any of the recited pharmaceutical compositions by topical application, transdermal, buccal, oral, gavage, and injection or intravenous.
  • the present disclosure also provides for methods for reducing
  • microorganisms on any surface or liquid comprising contacting a surface or liquid with any of the recited compositions.
  • the present disclosure also provides for compositions and methods for reducing microorganisms in foods comprising contacting a food with any of the recited compositions.
  • the invention also provides a method of killing methicillin resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa biofilm colonies comprising spraying a solution of up to 30% PCA in 70% isopropyl alcohol on a surface containing a biofilm.
  • the surface is a solid surface, porous or semi- porous, or a cloth-like surface.
  • a bandage or wound dressing having a solution of PCA or 2,4,6 trihydroxybenzaldehyde (246 THBA) or mixtures thereof impregnated thereon, and methods of preparing same.
  • the invention provides a method of treating a solid surface, porous or semi-porous, or a cloth-like surface to sanitize, sterilize, reduce bacterial growth or inhibit growth of microorganisms, the method comprising treating the solid surface with a solution of PCA or 2,4,6 trihydroxybenzaldehyde (246 THBA) or mixtures thereof, or in the case of porous or semi-porous surface or cloth-like surface, treating with a solution of PCA or PCA crystals.
  • a solution of PCA or 2,4,6 trihydroxybenzaldehyde (246 THBA) or mixtures thereof or in the case of porous or semi-porous surface or cloth-like surface, treating with a solution of PCA or PCA crystals.
  • a method of treating a medical device or surgical implant to sanitize, sterilize, reduce bacterial growth or inhibit growth of microorganisms comprising treating the medical device or surgical implant with a solution of PCA or 2,4,6 trihydroxybenzaldehyde (246 THBA) or a mixture thereof.
  • An embodiment of the invention includes a method of treating skin of a mammal to kill microorganisms on the skin, the method comprising treating the skin with a solution of PCA or 2,4,6 trihydroxybenzaldehyde (246 THBA) or a mixture thereof to kill microorganism on the skin.
  • the skin can be treated before a surgical procedure or after a surgical procedure.
  • Another embodiment of the invention provides a method of killing P. acnes on the skin of a patient, the method comprising applying a solution of PCA or 2,4,6 trihydroxybenzaldehyde (246 THBA) or a mixture thereof to the skin of the patient to kill the P. acnes.
  • the invention also includes a solution comprising 20-30% PCA, and essential oils; as peppermint or lemon oil etc., propylene glycol and isopropyl alcohol.
  • essential oils as peppermint or lemon oil etc.
  • propylene glycol and isopropyl alcohol are essential oils.
  • liposomes may be used in dermal applications with the compositions of the invention.
  • the present invention also provides a covering or bandage for administering antimicrobial compositions to a wound, wherein the antimicrobial compositions is an element of the covering or bandage.
  • Methods for promoting healing of a wound and more specifically to methods and compositions including the application of an anthocyanin or an anthocyanidin or metabolites thereof in the form of a covering or bandage for promoting wound healing by reducing or preventing microbial growth and inducing the activation of growth hormones are described.
  • Pharmaceutical compositions containing anthocyanin or anthocyanidins or metabolites thereof suitable for transdermal administration to a mammal for promoting or inducing wound healing are also described.
  • Figure 1 provides the minimum, maximum and optimum pH for growth of microorganisms. Acidic environments retard proliferation of various bacteria.
  • Anthocyanins, anthocyanidins and main metabolites are unstable relative to basic pH; thus, anthocyanins, anthocyanidins and main metabolites thereof have the potential to lower the pH of wound tissue as well as any surfaces and act as bactericidal or bacteriostatic.
  • Figure 2 is the metabolic pathway of cyanidin-3-glucoside (C3G) and includes the chemical structures of cyanidin-3-glucoside and cyanidin and their metabolites.
  • Figure 3 is the chemical structure of Protocatechuic acid (PCA), a dihydroxybenzoic acid, a type of phenolic acid. It is a major metabolite of antioxidant polyphenols found in certain plants, including green tea.
  • PCA Protocatechuic acid
  • Figure 4 compares concentrations of C3G and PCA to determine optimal effective concentrations. Bacterial burdens for P. aeruginosa were compared after treatment with C3G or PCA at 48 and 96 hours. A concentration of PCA 25 mM was effective to reduce the bioburden with statistical significance at 48 hours. C3G at 100 and 200 mM concentrations were effective at reducing the bioburden at 48 and 96 hours.
  • Figure 5 is a chart disclosing potential sources of PCA.
  • Figure 6 is a table providing a summary of the effectiveness of certain anthocyanins, anthocyanidins and a metabolite, including bactericidal or bacteriostatic activity. During this test, the purity, concentrations and molecular weight of these test substances (compounds) were known. The carrier was water and the dose was accurately calculated. Delphinidin limited growth against C. perfringens, S. aureus, and MRSA. Pelargonidin limited growth of P. acnes, C.perfingens, S. aureus, MRSA, and S. pyogenes. Cyanidin CI was effective against C. difficile, C prefringens, S. aureus ATCH 6538, S.
  • PCA P. aeruginosa.
  • PCA was also effective on C. albicans, which is important considering its ability to form biofilms and difficulty in treating C. albicans when existing with a catheter or implant.
  • Figure 7 is a table summarizing in vitro test results of 2,4,6
  • Trihydroxybenzaldehyde and demonstrating its ability to act as an antimicrobial, including as a bactericidal or bacteriostatic. Specifically, 2,4,6 Trihydroxybenzaldehyde was effective against E. coli, K. pneumonia, P. aeruginosa, S. aureus 6538 and 33591 (MRSA); further it was effective against a fungi, Aureobasidium pullulans, ATCC 15233.
  • Figure 8A is a photographic image illustrating in vitro test results of 2,4,6 Trihydroxybenzaldehyde against P. aeruginosa.
  • Figure 8B is a photographic image illustrating in vitro test results of 2,4,6 Trihydroxybenzaldehyde against S. aureus 33591 (MRSA).
  • Figure 8C is a photographic image illustrating in vitro test results of 2,4,6 Trihydroxybenzaldehyde against P. acnes.
  • Figure 9A shows the results of a rodent back skin tape study where concentrations of PCA and C3G in a vehicle of water were utilized to determine effectiveness against P. aeruginosa skin infections.
  • Figure 9B shows the results of rodent back skin tape study where concentrations of PCA and C3G in a vehicle of water were utilized to determine effectiveness against P. aeruginosa skin infections.
  • Figure 10A shows the results of an additional rodent back skin tape study to determine effective dosages of PCA and C3G in a vehicle of water that would be bactericidal for P. aeruginosa.
  • Figure 10B shows the results of an additional rodent back skin tape study to determine effective dosages of PCA and C3G in a vehicle of water that would be bactericidal for P. aeruginosa.
  • Figure 10C shows the results of an additional rodent back skin tape study to determine effective dosages of PCA and C3G in a vehicle of water that would be bactericidal for P. aeruginosa.
  • Figure 1 1 A shows the results of a rodent back skin study to determine the effects of PCA and C3G on the local growth hormones in untreated skin wounds of rodents.
  • a concentration of 25 mM PCA increased local growth hormone levels of IGF-1 at the site of the untreated skin wound.
  • Figure 1 1 B shows the results of a rodent back skin study to determine the effects of PCA and C3G on the local growth hormones in untreated skin wounds of rodents.
  • a concentration of 25 mM PCA increased local growth hormone levels of TGF-Beta at the site of the untreated skin wound.
  • Figure 1 1 C shows the results of a rodent back skin study to determine the effects of PCA and C3G on the local growth hormones in untreated skin wounds of rodents.
  • a concentration of 25 mM PCA increased local growth hormone levels of EGF at the site of the untreated skin wound.
  • Figure 12A is a photographic image of rodents treated with a topical solution of C3G (28%); at an acidic pH, this solution maintains a purple or red color and quickly metabolized at elevated pH levels, the C3G changes to a pink or even clear color.
  • the purple color of C3G remained on the rodent wound surface scar, thus indicating the pH remained acidic on the wound surface.
  • the C3G material on the surface was confirmed by subsequent histology.
  • Figure 12B is a photographic image of tissue from a study utilizing the homogenized wound tissue method used in this study, whereby the purple color indicates that the wound probably retained an acidic pH.
  • Figure 13A shows the IGF-1 response to 25 mM PCA in various environments, including tape stripped, tape stripped and infected with P. aeruginosa, tape stripped and treated with PCA, and tape stripped infected with P. aeruginosa, and PCA treated.
  • Figure 13B shows the TGF- ⁇ response to 25 mM PCA in various environments, including tape stripped, tape stripped and infected with P. aeruginosa, tape stripped and treated with PCA, and tape stripped infected with P. aeruginosa, and PCA treated.
  • Figure 13C shows the EGF response to 25 mM PCA in various environments, including tape stripped, tape stripped and infected with P. aeruginosa, tape stripped and treated with PCA, and tape stripped infected with P. aeruginosa, and PCA treated.
  • Figure 13C shows the EGF response to 25 mM PCA in various
  • Figure 14 is a photographic image of a cross section of rodent skin.
  • Figure 15 is a photographic image of a cross section of rodent skin.
  • Figure 16 is a photographic image of a cross section of rodent skin.
  • Figure 17 is a photographic image of a cross section of rodent skin.
  • Figure 18 is a photographic image of a cross section of rodent skin.
  • Figure 19 is a photographic image ol " a cross section of rodent skin.
  • Figure 20 is a photographic image ol " a cross section of rodent skin.
  • Figure 21 is a photographic image ol " a cross section of rodent skin.
  • Figure 22 is a photographic image ol " a cross section of rodent skin.
  • Figure 23 is a photographic image ol " a cross section of rodent skin.
  • Figure 24 is a photographic image ol " a cross section of rodent skin.
  • Figure 25 is a photographic image ol " a cross section of rodent skin.
  • Figure 26 is a photographic image ol " a cross section of rodent skin.
  • Figure 27 is a photographic image ol " a cross section of rodent skin.
  • Figure 28 is a photographic image ol " a cross section of rodent skin.
  • Figure 29 is a photographic image ol " a cross section of rodent skin.
  • Figure 30 is a photographic image ol " a cross section of rodent skin.
  • Figure 31 is a photographic image ol " a cross section of rodent skin.
  • Figure 32 provides a chart studying t he effectiveness of anthocyanin anthocyanidin metabolites against various microbes, including P. acnes, C. difficile, E. coli 8739 and 43895, S. Aureus 6538, 33591 , P. Aeruginosa 9027, MRSA 51625 and
  • MRSE methicillin resistant staphylococcus epidermis
  • Figure 33 provides a chart summarize results of testing PCA against
  • Figure 34 provides a chart summarize results of testing PCA against
  • Figure 35 provides a chart summarize results of testing PCA against
  • Figure 36 provides a chart summarize results of testing PCA against
  • Figure 37 provides a chart summarize results of testing PCA against
  • Figure 38 provides a chart summarize results of testing PCA against
  • Figure 39 provides a chart of results of testing PCA against P. acnes on the skin.
  • Figure 40 shows the results for a single spray of 30% PCA in isopropyl alcohol on 10 million biofilms colonies of Pseudomonas aeruginosa.
  • Figure 41 shows the concentration of 30% has lesser effect on MRSA, but still 90%.
  • Figure 42 provides a composite of the results based only for no growth cultures following treatment by each solution. * Note that the 1 % PCA was not included.
  • Figure 43 provides the summation percentages of "no growth.”
  • Ranges can be expressed herein as “approximately” or from “about” one particular value, and/or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value.
  • a weight percent of a reagent, component, or compound unless specifically stated to the contrary, is based on the total weight of the reagent
  • reduce or other forms of the word, such as “reducing” or “reduction,” is meant decrease or lower a characteristic (e.g., inflammation, growth or viability of microorganisms).
  • promote or other forms of the word, such as “promoting,” is meant to induce a particular event or characteristic, or delay the development or progression of a particular event or characteristic, or to minimize the chances that a particular event or characteristic will occur.
  • Treating or other forms of the word, such as “treating,” “treatment” or treated,” is used here to mean to administer a composition or to perform a method in order to induce, reduce, eliminate, and prevent a characteristic (e.g., inflammation, growth or viability of microbes). It is generally understood that treating involves providing an effective amount of the composition to the mammal or surface for treatment.
  • vehicle or "vehicle carrier” as used herein refers to mean the manner in which the reagents or compositions may be delivered, including as a liquid, salve, soap, foam, cream, solution, gel, spray, powder, wipes, antibacterial treatments, wipes and the like.
  • wound or “wound associated condition” as used herein refers to a medical condition when the integrity of any tissue is compromised (e.g., burns, skin breaks, bone breaks, muscle tears, tendon injuries punctures, surgical incision sites, microdermabrasion site, skin graft site, ).
  • a wound may be caused by any act, infectious disease, underlying condition, fall, or surgical procedure.
  • a wound may be chronic, such as skin ulcers caused by diabetes mellitus, or acute, such as a cut or puncture from a sharp object, an animal bite or a gunshot.
  • growth factors include but are not limited to, fibroblast growth factor (FGF), FGF-1 , FGF-2, FGF-4, platelet-derived growth factor (PDGF), insulin-binding growth factor (IGF), IGF-1 , IGF-2, epidermal growth factor (EGF), transforming growth factor (TGF), TGF- ⁇ , TGF-a, and collagen growth factors, and/or biologically active derivatives of these growth factors.
  • FGF fibroblast growth factor
  • FGF-1 FGF-1
  • FGF-2 FGF-4
  • PDGF platelet-derived growth factor
  • IGF insulin-binding growth factor
  • IGF-1 insulin-binding growth factor
  • IGF-2 insulin-binding growth factor
  • IGF-2 insulin-binding growth factor
  • EGF epidermal growth factor
  • TGF transforming growth factor
  • TGF- ⁇ TGF- ⁇
  • TGF-a and collagen growth factors
  • bacteria or “antimicrobial” is meant the ability to effect (e.g., eliminate, inhibit decrease, or prevent) microorganism growth, viability, and/or survival at any concentration. It also means kill the microorganism.
  • bacteriostatic is meant the ability to effect (e.g., stabilize or prevent future growth or prevent new growth) microorganism growth at any concentration.
  • a bacteriostatic compound, agent or reagent does not eliminate or kill the bacteria.
  • additive or “food additive” is meant to the use as a component of any food (including any substance intended to use in producing manufacturing, packing, processing, preparing, treating, packaging, transporting, or holding food).
  • antiseptic is meant an antimicrobial reagent or composition that is applied to any surface, including skin or tissue, to effect (e.g., eliminate, inhibit, decrease or prevent) microorganism growth, viability, and/or survival.
  • disinfect or other forms of the word, such as “disinfectant” or “disinfecting,” is meant decrease or lower a characteristic (e.g., eliminate, reduce, inhibit, decrease, or prevent) microorganism growth, viability or survival at any concentration. It is generally understood that disinfect involves providing an effective amount of the composition to any surface, but particularly solid surfaces, whether smooth or porous or semi-porous, or cloth-like surfaces.
  • sanitizing is meant decrease or lower a characteristic (e.g., eliminate, reduce, inhibit, decrease, or prevent) microorganism growth, viability or survival at any concentration. It is generally understood that sanitizing involves providing an effective amount of the composition to any surface. Further, it is generally understood that sanitizing solutions and sanitizing components are those solutions that may be safely used on food- processing equipment and utensils and on other food-contacting conditions.
  • sterilizize it is meant to kill microbes on the article being sterilized. Sterilize and sterilization include cold sterilization methods.
  • isolated or "an isolate” as it refers to either the compounds or reagents described herein means not 100% by weight but rather approximately 95% to 97% of the compound or reagent by weight.
  • alkyl as used herein is a branched or unbranched
  • hydrocarbon group of 1 to 20 carbon atoms Non limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, eicosyl, tetracosyl, and the like.
  • alkyl group can also be substituted or unsubstituted.
  • alkoxy or "alkyoxy group” as used herein refers to a branched or unbranched hydrocarbon chain having from 1 to 15 carbons and linked to oxygens. Non-limiting examples include methoxy, ethoxy and the like.
  • ProC3GTM commercially available ChromaDex®, Inc. Irvine, CA product
  • ChromaDex® Commercially available ChromaDex®, Inc. Irvine, CA product
  • the term "medicament” as used herein refers to any wound treatment, including but not limited to the group consisting of burn relief medications, anesthetic agents, wound cleansers, antiseptic agents, scar reducing agents, immunostimulating agents, antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, acne treating agents, sunscreen agents, dermatological agents, antihistamine agents, antibacterial agents, bioadhesive agents, inhibitors of prostaglandin synthesis, antioxidants, and mixtures thereof.
  • the term "nutraceutical” as used herein refers to any food stuff, including a dietary supplement or fortified food, provided for potential health and medical benefits.
  • the present invention provides methods, compositions and uses for treating and promoting healing of a wound. More specifically, the methods and compositions described herein include the administration of an anthocyanin or an anthocyanidin or metabolites thereof, preferably PCA and 2,4,6
  • Trihydroxybenzaldehyde (2,4,6 THBA) for promoting wound healing by reducing or preventing microbial growth and inducing the activation and or optimization of growth hormones.
  • the methods and compositions are used for the treatment of mammals, including humans. As with humans, there is a need for new antimicrobial compositions for the treatment of animals, including equine, canine and feline, due to resistance or allergic reactions to current antimicrobial compositions or agents. Therefore, the methods and compositions disclosed herein will be useful for the treatment and promotion of wound healing in livestock as well as domestic pets and will have broad- spectrum activity against microbes.
  • the present invention provides a broad spectrum antibiotic.
  • the inventor has shown the PCA and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA) are a broad spectrum antibiotic.
  • the inventor has shown that the parent anthocyanins or an anthocyanidins had limited antibiotic properties including cyanidin chloride. Further the subsequent acids in the metabolic chain; vanillic and hippuric acid had little activity as well. The inventor has thus identified the specific metabolites in the chain that had the antibiotic properties.
  • Methods and compositions described herein include the administration of an anthocyanin or an anthocyanidin or metabolites thereof, preferably PCA and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA) as a skin sterilizer (antiseptic).
  • an anthocyanin or an anthocyanidin or metabolites thereof preferably PCA and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA) as a skin sterilizer (antiseptic).
  • the normal human skin bacterial resident flora is one source of surgical site infections.
  • Two such commercially available preparations are Chloroprep® and
  • Betadine® Reports of their effectiveness in the medical literature show residual bacteria. Propionibacterium acnes is one such organism.
  • PCA Protocatechuic acid
  • 2,4,6 a phytochemical metabolite
  • Trihydroxybenzaldehyde (246 THBA) or mixtures thereof can be applied to the skin or wound in advance or after a surgical or dental procedure to kill bacterial present on the skin or wound. It has been shown (See figures 6 and 32) that PCA and 2,4, 6 THBA have the ability to kill a wide spectrum of microbes. For example, figure 6 provides the results of testing showing that protocatechuic acid (PCA), the main metabolite from anthocyanins and anthocyanidins, was effective against all bacteria tested as well as C. albicans and K. pneumonia. Importantly for skin wound treatment, PCA was effective against S. aureus 6538 and 33591 (MRSA) and P. aeruginosa. PCA was also effective on C.
  • PCA protocatechuic acid
  • MRSA MRSA
  • P. aeruginosa P. aeruginosa
  • MRSE staphylococcus epidermis
  • Figure 32 shows that 2,4,6 THBA is effective against E. coli 8739 and 43895, S. Aureus 6538, S. Aureus 33591 , P. Aeruginosa 9027, methicillin resistant staphylococcus epidermis (MRSE), including MRSE ATCC 51625, and Legionella 43662, and others.
  • MRSE methicillin resistant staphylococcus epidermis
  • the compositions promote wound healing by reducing or preventing microbial growth and inducing the activation and or optimization of local skin growth hormones.
  • the present invention also provides methods, compositions and uses for treating surfaces (solid, smooth, porous or semi-porous, or cloth-like) and liquids to reduce microbial growth or to sanitize or sterilize the surface. More specifically, the methods and compositions described herein include contacting any surface with a composition comprising an anthocyanin or an anthocyanidin or metabolites thereof (preferably PCA and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA)) thereby reducing or preventing microbial growth on said surface, or to sanitize or sterilize the surface.
  • the surfaces can be in the health care setting, sports setting or even food preparation settings or any setting where sterile surfaces are required.
  • compositions can be applied to solid surfaces such as implants, or solid surfaces like operating tables, benches, equipment, patient beds, etc. or surgical instruments to sanitize or sterilize the surface.
  • solid surfaces such as implants, or solid surfaces like operating tables, benches, equipment, patient beds, etc. or surgical instruments to sanitize or sterilize the surface.
  • the implants can be treated and coated with the compositions before inserted into the patient.
  • the instruments may be treated before use on a patient.
  • Solid surfaces such as operating tables, other equipment and other surfaces can be treated as well by spraying of the surface with compositions comprising PCA, 2,4,6 THBA or mixtures thereof.
  • compositions can be applied to smooth, porous or semi- porous, or cloth-like surfaces such as wound dressings, bedding, vascular implants, bandages, etc.
  • the material can be treated with the PCA solution and then used immediately or the material can be allowed to dry and then used.
  • a bandage can be treated with PCA and then allowed to dry and store (for about up to 2 years known shelf life).
  • the bandage can either be applied directly to the wound or can be wetted with water, 70- 90% isopropyl alcohol, saline or propylene glycol and/or essential oils and then applied to the wound. It is preferred to use propylene glycol and an essential oil as they enhance the absorption of PCA into the skin.
  • the antimicrobial composition of this invention When the antimicrobial composition of this invention is applied to as an element of a covering or bandage, to adhere to a surface to be treated, such as a wound, the antimicrobial composition generally can include a concentration of the PCA or 246 THBA or mixtures thereof of at least 1 .24% and 30% by weight of the compound depending upon the chemical nature of the vehicle, the target being treated and the species of bacteria to be treated. In certain embodiments a 25mM concentration of PCA is applied to the bandage. In Trans Pharm's independent laboratory experiment 367 on tape stripped wounds or rodents, the data demonstrated that C-3-G at 100 and 200 mM significantly decreased the bacterial burden of P. aeruginosa in skin tissue at 48 and 96 hours.
  • the isolated anthocyanins, anthocyanidins, or metabolites compounds this invention will be between 20-30% by weight of the compound for one intended use and more preferably, between 1.24% and 30% by weight of the compound depending upon the chemical nature of the vehicle, the target being treated and the species of bacteria to be treated.
  • the antimicrobial composition comprises PCA or 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA) or mixtures thereof. These bandages can promote wound healing not only as they are effective against killing microbes, but they also promote healing by inducing the activation and or optimization of local skin growth hormones.
  • the methods and compositions described herein include adding the composition comprising an anthocyanin or an anthocyanidin or metabolites thereof to liquid or fluid, including other sanitizing solutions and/or sanitizing components, thereby reducing or preventing microbial growth on said surface. Further still, the methods and compositions described herein include adding the composition comprising an anthocyanin or an anthocyanidin or metabolites thereof to any other vehicle, including but not limited to a powder, paste, cream foam, gel, wipes, other sanitizing components and the like thereby reducing or preventing microbial growth on said surface.
  • compositions for, and methods of destroying, killing or significantly reducing a bacterial biofilms are comprised of bacteria that form colonies and produce a surrounding matrix film to protect themselves.
  • the biofilm forming bacteria can form colonies that attach to foreign bodies, each other and tissues.
  • the bacteria aggregate in clusters and are surrounded by extracellular polymer matrix.
  • the biofilms are hard to destroy and therefore kill the underlying bacteria and provide the basis for much of the antibiotic resistance that has developed.
  • Biofilms can be found attached to surfaces such as implants and catheters and they also can be embedded in the biological host, such as for example cystic fibrosis wounds.
  • MRSA biofilms play a role in many device-related infections such as native valve endocarditis, otitis media, urinary tract infections, cystic fibrosis, acute septic arthritis, total joint implantation, catheters, pacemakers, etc.
  • the formation of a biofilm is a two-step process: 1 . Adherence of cells to a foreign body surface; and 2.
  • the present invention thus provides a composition that destroys biofilms.
  • the composition is preferably PCA.
  • the PCA may be mixed with 70% isopropyl alcohol and or small amount of essential oil; i.e. lemon, peppermint, etc.
  • the concentration of PCA can be anywhere from about 20% by weight PCA to 100% PCA.
  • the concentration of PCA varies with the intended purpose from 1 .24%, 20%, 30%, 20-50% or 20-40% or 20-30% by weight.
  • the crystals can be up to 100% PCA by weight.
  • the PCA may be in the form of crystals that are embedded into a material such as a cloth or a mesh, such as titanium or stainless steel. The crystals are applied to the metal where there is surface
  • the inventor has shown that a composition comprising PCA was able to stop the formation of a biofilm as well as kill bacterial in already formed biofilms.
  • the biofilms tested were Pseudomonas Aeruginosa ATCC 700888 and Staphylococcus aureus ATCC 33591 (MRSA). The tests are described in more details below and in Examples 7 and 8.
  • anthocyanidin, and or the main metabolites, PCA and or 246 THBA vary with the application and the vehicle.
  • the dose used on an open rodent taped stripped skin would be conditioned by the wound's tolerance of the vehicle which in this case would be water, normal saline or like.
  • the amount of PCA for instance that can be dissolved in water is 1 .24 grams per 100 ml.
  • the effective dose of C-3-G and PCA were expressed in micromolars (mM) as 25 to 200 mM dose. Conversion factors are necessary to make comparisons to larger doses used in other applications. 78 mM (mmole/L) PCA would be 12.021 g PCA per Liter.
  • Propionibacterium acnes Note that there was no skin penetration properties to water by this method.
  • the increase of PCA concentration was possible to as much as 17% when in composition of matter was 70% isopropyl alcohol with propylene glycol and essences of peppermint oil.
  • This composition had skin penetration properties.
  • the amount of liquid applied was no more than one milliliter by cotton swab. Therefore the topical dose on intact human skin was 1 .7 grams (estimated). This is many times that required for traumatized taped stripped skin.
  • PCA necessary for coating metal and or cloth was 20 to 30% PCA or 20-30 grams per 100 ml of 70% isopropyl alcohol. These compositions allowed for higher concentrations, and also evaporated rapidly to dry state of PCA crystals on the metal or cloth.
  • Methicillin resistant Stapylococcus aureus required higher dose than Pseudomonas aeruginosa. See figure 35. However at concentration of 30%, it was effective. See figure 36. The application on an implant allowed to dry had the above results. However, when a glass surface is covered with 10 million biofilms colonies the results differ with the concentration and the bacteria biofilms to be eradicated.
  • the glass surface experiments are described in example 8. Generally, Glass slides were inoculated at Time 0. Batch phase was performed for 6 hours to allow for biofilm formation on the glass slides. The drip flow mechanism was then turned on to provide a continuous flow of nutrients to the glass slides over 48 Hours. After 48 hours, 2 sets of glass slides were sprayed with a 30% PCA solution.
  • compositions of the present invention show discovery of dose and composition requirements for any given intended use related to nature of the wound, the disinfectant intentions, environmental conditions of coating and or spraying plus consideration of the bacterial species and biofilms to be destroyed.
  • these compositions also have broad spectrum activity against a wide range of microbes.
  • these compositions may also be used in combination with other wound treatments, including other antimicrobials.
  • the additional antimicrobial is not sulfamethoxazole.
  • anthocyanins and anthocyanidins or combinations of anthocyanins, anthocyanidins or their metabolites that are bactericidal or antimicrobial was determined by conducting in vitro testing.
  • Anthocyanidins that were tested at 100 mM (44.938 grams per liter) with less than one milliliter per dose included delphenindin, pelargonindin, and cyanidin CI and cyanidin-3-glucoside.
  • Protocatechuic acid and 2,4,6 trihydroxybenzaldehyde, the anthocyanidin metabolites, were also tested at the same concentrations. Referring to FIGS. 6-8, delphinidin limited growth against C.
  • Cyanidin CI was effective against C. difficile, C prefringens, S. aureus ATCH 6538, S. aureus (MRSA) ATCH 33591 , S.
  • PCA Protocathechuic acid
  • MRSA MRSA
  • K. pneumoniae the main metabolite from anthocyanins and anthocyanidines
  • PCA was effective against all bacteria tested as well as C. albicans and K. pneumonia.
  • S. aureus 6538 and 33591 MRSA
  • P. aeruginosa was also effective on C. albicans, which is important considering its ability to form biofilms and difficulty in treating C.
  • CFU colony forming units
  • anthocyanins, or anthocyanidins, or metabolites thereof are provided in any recited composition or method of use in a range of between 20 to 200 mM. In yet other embodiments, the anthocyanin, anthocyanidin, or metabolites thereof provided in any recited composition or method of use is provided in a range of between 20 to 100 mM. In yet other embodiments, the anthocyanin, anthocyanidin, or metabolites thereof provided in any recited composition or method of use is provided in a range of between 20 to 50 mM. In a preferred embodiment, anthocyanins or anthocyanidins and or their metabolites are provided in concentrations of about 100 mM or less.
  • bacteria have a range of pH at which growth is optimized, and most bacteria are more viable at basic pH ranges. Generally,
  • anthocyanins, anthocyanidins and their metabolites also have an acidic pH and have the potential to have bactericidal or bacteriostatic modes of action. Because C3G and PCA reagents have an acidic pH, their bactericidal or bacteriostatic mode of action is by direct contact with the bacteria.
  • Anthocyanins and anthocyanidins were further studied to determine effects on wound healing, including whether they had any effect on the optimization of local growth hormone activity at the wound site along with other supporting histological evidence of promoting healing.
  • Examples of common local growth hormones related to skin wound healing include Epidermal growth factor (EGF), Insulin-like growth factor-1 (IGF-1 ) and Transforming Growth Factor -Beta (TGF- ⁇ ).
  • EGF Epidermal growth factor
  • IGF-1 Insulin-like growth factor-1
  • TGF- ⁇ Transforming Growth Factor -Beta
  • Epidermal growth factor or EGF is a growth factor that stimulates cell growth, proliferation, and differentiation by binding to its receptor EGFR.
  • IGF-1 is important in skin repair by stimulating keratinocyte proliferation and migration as well as collagen production by fibroblasts. Its expression is important during wound healing such that retarded healing has been correlated with reduced IGF-1 levels. While local administration of IGF-1 to wound sites enhanced wound closure and stimulated granulation tissue formation, increased IGF-1 receptor expression was reported in chronic wounds and in hypertrophic scars. Additionally, IGF- 1 stimulation was associated with increased invasive capacity of keloid
  • TGF- ⁇ also is important in skin would healing; however, it is considered a pro-fibrotic growth factor and increased levels of TGF- ⁇ or prolonged presence has been identified as causing hypertrophic scaring.
  • FIG. 1 tests were performed on rodent skin to explore the effects of PCA on the local growth hormones in rodent skin.
  • a concentration of 25 mM PCA increased local growth hormone levels at the site of the untreated skin wound.
  • figure 1 1 demonstrates that a single reagent or compound would optimize local growth hormones to promote healing without scarring.
  • Approximately 25 mM PCA was the optimal reagent and dose.
  • optimization is possible using the compositions of the present invention.
  • all three local growth hormones were lowered in the simulated clinical pathological environment (stripped and infected); however, the lowering of these hormones was not to the extent of absences.
  • the necessary IGF-1 is still above the controls in this environment; however, the scar forming properties of the other two hormones have been markedly reduced. Therefore, optimization of local growth hormones is achieved.
  • the optional concentration of PCA was confirmed as 25 mM PCA in this situation and environment, meaning local growth hormone growth levels were optimized at this dosage such that IGF-1 as moderately elevated while TGF- ⁇ and EGF levels were decreased. This is important to promote wound healing while preventing potential scarring.
  • the therapeutic effective dose may vary depending on a wide variety of factors. For instance, the dose may vary depending on the formulation, method of application of the therapeutic reagent or combination with other reagents, or
  • compositions, compounds or combination of compositions or compounds to the wound are compositions, compounds or combination of compositions or compounds to the wound.
  • a method of promoting healing of a wound in a mammal comprising administering an anthocyanin or an anthocyanidin to the mammal in need of such treatment a therapeutically effective amount of the anthocyanin or anthocyanidin compound wherein microbial growth is prevented or reduced and local growth hormone activity is optimized.
  • a method of promoting healing of a wound in a mammal comprising administering an anthocyanin metabolite or an anthocyanidin metabolite to the mammal in need of such treatment a therapeutically effective amount of the metabolite or an anthocyanidin metabolite compound wherein microbial growth is reduced and local growth hormone activity is optimized.
  • a method of promoting healing a wound further comprises applying a wound dressing or bandage that has been treated with a composition of PCA or 246 THBA or mixtures thereof.
  • a method of treating P. acnes in a patient in need thereof comprising administering an anthocyanin metabolite to the patient in need of treatment in an therapeutically effective amount of protocatechuic acid wherein growth of P. acnes is reduced.
  • Figure 32 shows that both C-3-G and PCA were able to kill P. acnes, specifically P. acnes 6919.
  • a method of prophylactically treating a preoperative skin incision site comprising administering an anthocyanin, an anthocyanidin and/or a metabolite to a patient in need of such treatment an effective amount of the anthocyanin or anthocyanidin compound wherein microbial growth is prevented or reduced.
  • a method of disinfecting a surface comprising contacting said surface with an anthocyanin, an anthocyanidin and/or a metabolite thereof in an effective amount of the anthocyanin, anthocyanidin and/or metabolite compound wherein microbial growth is prevented, reduced or eliminated and, further, where the microbial growth that is reduced is methicillin resistant staphylococcus aureus (MRSA).
  • MRSA methicillin resistant staphylococcus aureus
  • a method of disinfecting or sterilizing a surface comprising contacting said surface with an anthocyanin, an anthocyanidin and/or a metabolite thereof or combination thereof in an effective amount of the anthocyanin, anthocyanidin and/or metabolite or combination thereof wherein microbial growth is prevented, reduced or eliminated and, further, where the microbial growth that is reduced may be an endogenous or exogenous source, including but not limited to P. acnes, S. aureus, P. aeruginosa, E. coli, S. epidermidis, S.
  • a method of post-operative treating a post-operative skin site comprising administering an anthocyanin, an anthocyanidin and/or a metabolite to a post-operative skin site, such as a skin graft, skin graft donor site, a microdermabrasion site, or a surgical incision site, in an effective amount of the anthocyanin, anthocyanidin and/or metabolite compound wherein microbial growth is prevented, reduced or eliminated and local growth hormone production is optimized.
  • a post-operative skin site such as a skin graft, skin graft donor site, a microdermabrasion site, or a surgical incision site
  • This disclosure also provides for a method comprising contacting a surface with an effective amount of the antimicrobial composition.
  • an effective amount of a composition as provided herein is meant an amount of a composition sufficient to provide the desired benefit, either bactericidal or bacteriostatic (e.g., reduction or prevention of microorganism growth or survival).
  • the exact amount required will vary from use to use depending on a variety of processing parameters, as understood by one of ordinary skill, such as the type of surface, the type of microorganism to be treated, the surface size, the mode of deliver (e.g., aerosol, spraying or dipping), and the like. Determination of what constitutes an "effective amount” is made by routine testing with known concentrations and adjusting those concentrations as needed to obtain the desired benefit and can be determined by one of ordinary skill in the art using only routine experimentation.
  • the antimicrobial composition of this invention When the antimicrobial composition of this invention is applied to a surface to be treated, the antimicrobial composition generally can include a
  • concentration of the anthocyanins and anthocyanidins of at least 25 mM concentration, not including the carrier.
  • the isolated anthocyanins, anthocyanidins, or metabolites compounds this invention will be between 90 %-97% by weight of the compound, and more preferably, between 95% - 98% by weight of the compound.
  • compositions of this invention When the antimicrobial composition or compositions of this invention are applied to a surface to be treated may be diluted for use as a sanitizer or as a
  • a method of inhibiting growth of a biofilm on a solid, smooth, porous or semi-porous, or cloth-like surface by treating the surface with a composition of the present invention, preferably comprising PCA.
  • the surface can be any solid, smooth, porous or semi-porous, or cloth-like surface, including implants that are inserted into a patient.
  • antimicrobial compositions Disclosed herein, in one aspect, are antimicrobial compositions.
  • the disclosed antimicrobial reagents and compositions can be used to eliminate, reduce, and/or prevent microorganism growth, viability, or survival.
  • the present invention thus provides a composition that destroys or inhibits growth of a biofilm.
  • the composition is preferably PCA.
  • the PCA may be mixed with 70% isopropyl alcohol.
  • the concentration of PCA can be anywhere from about 20% PCA to 100% PCA.
  • the concentration of PCA is about 20-50% by weight, or is about 20-40% by weight or is about 20-30% by weight or is 30% to 50% by weight.
  • the PCA may be in the form of crystals that are embedded into a material such as a cloth or a mesh, such as titanium or stainless steel.
  • PCA in water only decreased the colonies of P. acnes in and on human skin as the PCA concentration he was able to achieve when dissolving in water was limited to 1.24%.
  • the use of isopropyl alcohol in the combination was not an obvious choice because isopropyl alcohol alone only is marginally effective in killing bacteria.
  • the invention provides a composition comprising about 17 to 40%, or 17 to 30% or 17 to 20 % by weight of PCA, isopropyl alcohol, propylene glycol and an essential oil, preferably of peppermint, or a citrus fruit (i.e. lemon, grapefruit, orange, lime, etc.).
  • composition is useful in the methods described in the invention, for example as a skin antiseptic as a surface disinfectant, as a spray to disinfect a surface, etc. Further, the inventor found that neither propylene glycol or essential oil alone or in combination absent the isopropyl alcohol provide enough concentration of PCA to be above the effective range of 10% concentration. Therefore the composition of the invention, to be effective should have at least PCA at 17+% by weight in at least 70-90% isopropyl alcohol, propylene glycol (15 mis in a 105 ml total solution) and the essential oil; i.e. peppermint or lemon etc.
  • the invention further provides a composition of PCA wherein the composition comprises or consists of PCA that can be applied directly or provided in various vehicles depending upon the application.
  • PCA 1.24 grams in 100 milliliters of water was effective on open wound.
  • a composition of 70% isopropyl alcohol, propylene glycol and essential peppermint oil was also effective in use as a skin antiseptic, especially against P. acnes.
  • Studies showed that higher concentration of 10% PCA (20 grams in 90 milliliters of 70% isopropyl alcohol) was more effective than PCA in water.
  • concentration was more effect than just PCA in isopropyl alcohol - the composition consisted of PCA (20 grams) 70% isopropyl alcohol (85ML), propylene glycol (15 ml) and an essential oil (5 ml).
  • the invention also provides bandages, wound dressings and the like comprising PCA or 246 THBA.
  • the disclosed antimicrobial composition can be selected from the list of anthocyanins, anthocyanidins, metabolites of anthocyanin and anthocyanidin metabolites, or a combination thereof.
  • the anthocyanin can be selected from cyanidin-3-glucoside or delphinidin-3- glucoside, cyanidin-3-galactoside, and pelargonidin-3-galactoside.
  • the anthocyanidins can be selected from cyanidin, delphinidin, pelargonidin, malvidin and petunidin.
  • metabolites can be selected from
  • the present invention provides for a pharmaceutical composition for promoting wound healing, comprising phytochemicals: a) an
  • anthocyanin or b) anthocyanidin or their metabolites such as C3G, PCA, 246 THBA, vanillic and hippuric acid.
  • the anthocyanin can be selected from cyanidin-3-glucoside or delphinidin-3-glucoside, cyanidin-3-galactoside, and
  • the anthocyanidins can be selected from cyanidin, delphinidin, pelargonidin, malvidin and petunidin.
  • the present composition comprises PCA and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA).
  • the present disclosure provides for a pharmaceutical composition comprising protocatechuic acid (PCA) whereby said composition reduces the growth of certain microbes, including P. acnes.
  • PCA protocatechuic acid
  • the present disclosure provides for a pharmaceutical composition comprising cyanidin-3-glucoside whereby said composition reduces the growth of certain microbes, including H. pylori.
  • the present invention provides for a pharmaceutical composition for treating a wound, comprising: a) an anthocyanin; b) anthocyanidin; or c) a metabolite of an anthocyanin or anthocyanidin in an effective amount whereby microbial growth is reduced.
  • a pharmaceutical composition for treating a wound comprising: a) an anthocyanin; b) anthocyanidin; or c) a metabolite of an anthocyanin or anthocyanidin in an effective amount whereby microbial growth is reduced.
  • the present composition comprises PCA and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA).
  • the anthocyanin can be selected from cyanidin-3- glucoside or delphinidin-3-glucoside, cyanidin-3-galactoside, and pelargonidin-3- galactoside.
  • the anthocyanidins can be selected from cyanidin, delphinidin, pelargonidin, malvidin and petunidin.
  • metabolites can be selected from protocatechuic acid (PCA) and 2, 3, 4 trihydroxybenzaldehyde.
  • the pharmaceutical composition of this invention to treat a wound generally can include a concentration of the anthocyanins and
  • the present composition comprises PCA and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA).
  • the pharmaceutical composition of this invention to treat a wound can include a concentration of the anthocyanins and anthocyanidins or metabolites thereof in a concentration of between 20 mM to 200 mM concentration, not including the carrier.
  • the pharmaceutical composition of this invention to treat a wound can include a concentration of the anthocyanin,
  • the pharmaceutical composition of this invention to treat a wound can include a
  • PCA in a pharmaceutical composition of this invention, can be provided in a concentration approximately 50-100 mM.
  • pterostilebene can be provided in a concentration of approximately 35-65 mM.
  • PCA can be provided in a
  • PCA and psterostilbene can be provided in a combination in concentrations provided in this disclosure.
  • C3G would be provided at a dosage of 131 , 261 and 522 mg/kg.
  • the present invention provides for a pharmaceutical composition for treating a wound, comprising phytochemicals: a) an anthocyanin; b) anthocyanidin; c) a metabolite of an anthocyanin or anthocyanidin such as C3G, PCA, 246 THBA, vanillic and hippuric acid.
  • the present composition comprises PCA and 2,4,6 Tnhydroxybenzaldehyde (2,4,6 THBA).
  • the present disclosure also provides for a pharmaceutical composition
  • a pharmaceutical composition comprising phytochemicals: a) an anthocyanin; b) anthocyanidin; c) a metabolite of an anthocyanin or anthocyanidin such as C3G, PCA, 246 THBA, vanillic and hippuric acid.
  • the present composition comprises PCA and 2,4,6 Tnhydroxybenzaldehyde (2,4,6 THBA).
  • the present invention provides for a pharmaceutical composition for promoting wound healing, comprising phytochemicals: a) an
  • anthocyanin b) anthocyanidin; c) a metabolite of an anthocyanin or anthocyanidin such as C3G, PCA, 246 THBA, vanillic and hippuric acid.
  • metabolites can be selected from, protocatechuic acid, 2,4,6 trihydroxybenzaldehyde.
  • the present disclosure provides for a pharmaceutical composition comprising phytochemicals: a) an anthocyanin; b) anthocyanidin; c) a metabolite of an anthocyanin or anthocyanidin such as C3G, PCA, 246 THBA, vanillic and hippuric acid.
  • the metabolites are metabolites selected from protocatechuic acid, 2,4,6 trihydroxybenzaldehyde.
  • compositions whereby the anthocyanin, anthocyanidin, anthocyanin metabolite, anthocyanidin metabolite, anthocyanin metabolite, or metabolites thereof, are isolated reagents.
  • the present composition comprises PCA and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA).
  • the present disclosure also provides for routes of administration of the pharmaceutical compositions, including oral, injection, intravenous, topical, sublingual, buccal, inhalation, intradermal, subcutaneous, soft tissue, and cutaneous.
  • Oral administration of the compositions of this disclosure may include a liquid or semisolid form, tablet, pill, capsule, powder, or gel.
  • oral administration will be in a liquid composition.
  • Compositions including a liquid pharmaceutically inert carrier such as water may be considered for oral administration.
  • Other pharmaceutically compatible liquids or semisolids may also be used. The use of such liquids and semisolids is well known to those of skill in the art.
  • Intravenous and injection administration will be in liquid form.
  • Other pharmaceutically compatible liquids or semisolids may also be used.
  • the use of such liquids and semisolids is well known to those of skill in the art.
  • the composition is formulated as a topical composition.
  • the vehicle of the topical composition delivery is in the form of a liquid, salve, soap, spray, foam, cream, emollient, gel, ointment, balm or transdermal patch.
  • compositions can be in the form of an aqueous solution.
  • the compositions disclosed herein can also be in the form of a liquid, gel, suspension, dispersion, solid, emulsion, aerosol, for example, powders, tablets, capsules, pills, liquids, suspensions, dispersions or emulsions.
  • the compositions disclosed herein can be in the form suitable for dilutions.
  • the compositions can be in the form of a powder, cream, paste, gel or solid that can be reconstituted.
  • the antimicrobial composition can also include at least one additive selected independently from a carrier, a diluent, an adjuvant, a solubilizing agent, a suspending agent, a filler, a surfactant, a secondary antimicrobial agent, a preservative, a viscosity modifier, a thixotropy modifier, a wetting agent, an emulsifier, or any combinations thereof.
  • the disclosed antimicrobial composition can further comprise at least one surfactant selected from a cationic surfactant, an anionic surfactant, a non- ionic surfactant, and an amphoteric surfactant.
  • the disclosed antimicrobial and/or pharmaceutical compositions may further comprise medicament is selected from the group consisting of burn relief medications, anesthetic agents, wound cleansers, antiseptic agents, scar reducing agents, immunostimulating agents, antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, acne treating agents, sunscreen agents, dermatological agents, antihistamine agents, antibacterial agents, bioadhesive agents, inhibitors of prostaglandin synthesis, antioxidants, and mixtures thereof.
  • the disclosed antimicrobial compositions can optionally include one or more additives such as carriers, adjuvants, solubilizing agents, suspending agents, diluents, surfactants, other antimicrobial agents, preservatives, fillers, wetting agents, antifoaming agents, emulsifiers, and additives designed to affect the viscosity or ability of the composition to adhere to and/or penetrate the wound.
  • additives such as carriers, adjuvants, solubilizing agents, suspending agents, diluents, surfactants, other antimicrobial agents, preservatives, fillers, wetting agents, antifoaming agents, emulsifiers, and additives designed to affect the viscosity or ability of the composition to adhere to and/or penetrate the wound.
  • the disclosed antimicrobial compositions including the selected active components, including the anthocyanins or anthocyanidins and metabolites thereof, are without causing significant undesirable biological effects or interacting in a deleterious manner with any of
  • compositions including the selected active components, including the anthocyanins or anthocyanidins and metabolites thereof, are provided as a nutraceutical and provided as a dietary supplement without causing significant undesirable biological effects or interacting in a deleterious manner with any of the other components of the foodstuff in which it is contained.
  • the antimicrobial composition provided herein including anthocyanins or anthocyanidins and metabolites thereof, are used in agricultural settings, including but not limited to nurseries, commercial farming, agricultural research facilities, residential gardens and produce processing facilities, and are applied to plants and trees to inhibit, reduce or substantially eliminate microbial bioburden as well as many fungal bioburden on plants, trees, and surfaces thereof, including leaf surfaces.
  • antiseptic compositions of the present invention are formulated for use in liquids, solutions, gels, soaps, creams, powders salves and other preparations designed for topical use as antiseptic agents, sprays, foams, antibacterial treatments, wipes and the like.
  • antiseptic compositions of the present invention are formulated as a hand antiseptic.
  • antiseptic compositions of the present invention are used in industrial settings such as in water treatment facilities, including swimming pools or water treatment plants, food preparation, including but not limited to poultry and fish processing facilities or produce handling and packaging settings to inhibit, reduce or substantially eliminate microbial bioburden, as well as many fungal bioburden.
  • industrial equipment and surfaces may be contacted with, or soaked in, the antiseptic compositions of the present invention.
  • sanitizing compositions of the present invention are formulated for use in liquids, solutions, gels, soaps, and other preparations designed for use as sanitizing agents, liquids, including sprays, foams, gels, soaps, sanitizing treatments, and the like when used as a sanitizing solution, including but not limited to, use in food processing facilities, including food-processing equipment and utensils, and on other food-contact articles.
  • sanitizing compositions of the present invention use in food processing facilities, including food-processing equipment and utensils, and on other food-contact articles are formulated to include any components generally recognized as safe for use in food processing facilities, including but not limited to, aqueous solutions containing potassium, sodium or calcium hypochlorite, a solution of hydrogen peroxide, an aqueous solution containing potassium iodide, sodium lauryl sulfate, sodium-toluenesulfonchloroamide, solutions containing
  • dodecylbenzensulfonic acid other acceptable detergents and the like.
  • the one or more of the additives can be an agent that is acceptable when used in or on foods and beverages and which can be consumed by a mammal (e.g., human, pet, livestock, etc.) along with the selected active components, including the anthocyanins or anthocyanidins and metabolites thereof, without causing significant undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained.
  • a mammal e.g., human, pet, livestock, etc.
  • compositions of the present invention including those compositions comprising: a) an anthocyanin; or b) an anthocyanidin; or c) a metabolite of an anthocyanin metabolite; or d) a metabolite of an anthocyanidin metabolite, or e) a combination thereof, are used in food processing, packing, manufacturing, handling, preparing, treating, transporting or holding as a food additive without causing undesirable effects or interacting in a deleterious manner.
  • protocatechuic acid can be used as an additive in meat, including the handling and processing, without causing undesirable effects or interacting in a deleterious manner with the meat.
  • compositions of the present invention including those compositions comprising: a) an anthocyanin; or b) an anthocyanidin; or c) a metabolite of an anthocyanin metabolite; or d) a metabolite of an anthocyanidin metabolite, or e) a combination thereof, are used in food processing, including cold sterilization of food containers, including bottles, without causing undesirable effects or interacting in a deleterious manner.
  • the antimicrobial compositions disclosed herein can further comprise a carrier.
  • carrier means a compound, composition, substance, or structure that, when in combination with a compound or composition disclosed herein, facilitates preparation, administration, delivery, effectiveness, or any other feature of the compound or composition.
  • carriers include water, isopropyl alcohol ethanol, polyols (propyleneglycol, polyethyleneglycol, glycerol, and the like), vegetable oils, and suitable mixtures thereof.
  • “Pharmaceutically acceptable carrier” means a compound, composition, substance, or structure that is useful in neither preparing a pharmaceutical composition which is generally safe, non-toxic, and neither biologically nor otherwise undesirable and includes that which is acceptable for veterinary as well as human pharmaceutical use.
  • the antimicrobial compositions disclosed herein can also comprise adjuvants such as preserving, wetting, emulsifying, suspending agents, and dispensing agents. Prevention of the action of other microorganisms can be accomplished by various antifungal agents, for example, parabens, chlorobutanol, phenol, and the like.
  • Suitable suspending agents can include, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, and the like.
  • the disclosed antimicrobial compositions can also comprise solubilizing agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl alcohol, benzyl benzoate, propyleneglycol, 1 ,3- butyleneglycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol, tetrahydrofur fury 1 alcohol, polyethyleneglycols and fatty acid esters of sorbitan or mixtures of these substances, and the like.
  • the additives can be present in the disclosed compositions in any amount for the individual anthocyanin or anthocyanidin compound components.
  • Example 1 Use of in vitro studies for antimicrobial susceptibility testing of anthocyanins, anthocyanidins, or metabolites and compounds thereof.
  • This example describes the method for testing the antimicrobial susceptibility of anthocyanins, anthocyanidins, or metabolites and compounds thereof.
  • the Kirby-Bauer method of disc diffusion was used for testing, following a standard set of procedures recommended by the NCCLS.
  • a set of discs saturated with either testing compounds or a control was placed on inoculated agar plates.
  • the plates were inoculated with organisms listed in the tables provided in FIG.6, including C difficile, P. acnes, C. prefringens, L. casei, C. albicans, E. coli, ATTC 8739 and ATCC 43895, S. aureus, S. mutans, S. pyogenes, P. aeruginosa and K.
  • control sample was amoxicillin, an antimicrobial with very effective broad spectrum antibiotic properties.
  • Samples included delphinidin, pelargonidin, cyanidin CI, 28% cyanindin-3-glucoside (C3G), protocatechuic acid (PCA) and 2,4,6 Trihydroxybenzaldehyde (2,4,6 THBA).
  • the testing samples had bactericidal and bacteriostatic activity against many of the organisms.
  • P. acnes an organism that is very difficult to treat, often requiring multiple current antibiotics for effective treatment, was susceptible to both C3G and PCA. Indeed, both of these test samples were bactericidal against P. acnes.
  • PCA was also effective against Staphlococcus aureus ATCC 33591 , known as Methacillin Resistant Staph Aureus (MRSA), Staphlococcus epidermidis ATCC 51625, known as Methacillin
  • MRSE Resistant Staph Epidermidis
  • PCA was also shown to have some effectiveness against Pseudomonas aeruginosa, a common pathogen in wounds, especially burns, as well as chronic lung infections. Amoxicillin, the control sample, had no effect on P. aeruginosa. Similarly, Candida albicans, frequently a co pathogen in wounds, was susceptible to PCA.
  • PCA was also shown to have some effectiveness against Pseudomonas aeruginosa, a common pathogen in wounds, especially burns. Amoxicillin, the control sample, had no effect on P. aeruginosa. Similarly, Candida albicans, frequently a co pathogen in wounds, was susceptible to PCA.
  • the present invention provides advantages over the prior art, including providing anthocyanin, anthocyanidin, their metabolites or combinations thereof to a wound to provide a reduction or elimination of bacteria. It is contemplated that the invention will also find use in the treatment of surfaces, including medical devices and medical implants, to reduce or eliminate bacteria.
  • Example 2 Use of mouse model to determine dose levels and intervals of test samples. Methods: [0192] Mice had back skin tape stripped and the stripped site (wound) was infected with P. aeruginosa (ACTA 9027). The test reagents were applied topically in an aqueous solution on the stripped site at two hours and daily for four days.
  • Cyanidin 3-glucoside (C3G), an anthocyanin, and its main metabolite PCA were formulated and tested at several doses.
  • the aqueous carrier was water.
  • the C3G formulation included 50 mM, 100 mM and 200 mM dose concentrations.
  • the PCA formulation included at 50, 100 and 200 mM dose concentrations.
  • Results were collected from the mice at day five. Both C3G and PCA decreased the bacterial burden; however, none were statistically significant. See figure 9A. There was a trend towards a decreasing concentration of PCA, with 50 mM being the most effective. The most effective dose of C3G was 100 mM. It is contemplated that because C3G degrades to PCA in this environment, the test results may indicate that C3G was not being tested alone, but rather was a combination of C3G and its metabolites, including a combination of C3G and PCA as the effective agents.
  • Example 3 Use of mouse model to further determine effective dose levels and dose intervals of test samples.
  • mice had back skin tape stripped and the stripped site (wound) was infected with P. aeruginosa (ACTA 27853).
  • the test reagents were applied topically in an aqueous solution on the stripped site at two hours and daily on day 1 , 2 and 3.
  • C3G, an anthocyanin and its main metabolite PCA were formulated and tested at several doses.
  • the aqueous carrier was water.
  • the C3G formulation included 100 mM and 200 mM dose concentrations and the PCA formulation included 25 and 50 mM dose concentrations.
  • Results were collected from the mice at day two and four. Both C3G and PCA decreased the bacterial burden at 48 and 96 hours. (See Figure 9B) . The most significant decrease of bacteria was observed at 25 mM of and 100 and 200 mM of C3G. Although PCA at 25 mM reduced the bacterial burden at both time periods, its activity was statistically significant at 48 hours. C3G at both 100 mM and 200 mM significantly reduced the bacterial burden at 48 and 96 hours.
  • Example 4 Use of a mouse model for wound healing.
  • mice were shaved but unstrapped and uninfected (normal rodent skin).
  • the test reagents were applied topically in an aqueous solution on the unstripped site at two hours and daily on day 1 , 2 and 3.
  • Testing reagents consisted of C3G and PCA formulated at one dose, 100 ⁇ in an aqueous solution.
  • Example 5 Use of mouse model to determine isolated effect of 25 mM solution of PCA in various environments.
  • the PCA test reagent was applied topically in an aqueous solution on the stripped site at two hours and 24 hours.
  • the testing reagents consisted of and PCA formulated at one dose, 25mM, in an aqueous solution.
  • Levels of IGF-1 , TGF- ⁇ , and EGF levels in the skin tissue at 48 hours were measured by ELISA. There were two control groups; the stripped skin and the stripped skin and infected.
  • the infected stripped skin showed the highest level with IGF-1 (statistically significant) and TGF- ⁇ . This is representative of tissue response to injury and infection; similarly, the EGF response was very inconsistent compared to the other two growth hormones.
  • EGF response levels were different than either IGF-1 or TGF- ⁇ . They were highest in the stripped and uninfected wound and lowest in the stripped, infected and treated wound. Therefore, the treatment optimized the amount of hormone production compared to the untreated infection. This is beneficial to limit scarring while promoting healing over the controls.
  • PCA at 25 mM acts on stripped and infected mice skin and optimizes the IGF-1 production and optimizes the local growth hormones.
  • Example 6 Use of mice to establish wound promoting effect of compositions.
  • Control Group 1 three mice with only tape stripped wounds on the back. These mice were not infected or treated. The skin was harvested at time zero, 2 and 48 hours for histology examination.
  • Control Group 2 three had tape stripped wounds and infection. Tissue submitted at 2 and 48 hours for histological examination.
  • mice had skin stripped wounds and infection. Treatment varied by reagent and dosage. Testing reagents included PCA at 25 at 25 and 50 mM and C3G at100 and 200 mM.
  • Inoculum count was estimated before inoculation by optical density and confirmed after inoculation by dilution and back count.
  • testing reagents were topically applied at 2 and 24 hours with 100 ⁇ _ of fluid spread over the wound.
  • Thickness The thickness of the dermal layer was observed.
  • Hair Follicles The hair follicles and the layer of surrounding cells were observed. Hair follicles presence is critically important to skin wound healing. (Gharzi A, Reynolds AJ, Jahoda CA. Plasticity of hair follicle dermal cells in wound healing and induction. Exp Dermatol. 2003Apr; 12 (2): 126-36). The dermal sheath surrounding the hair follicle has the progenitor cells for contributing fibroblasts for wound healing.
  • Vascularity Vascularity was observed, but an assessment of
  • Inflammation The presence of cellular infiltration was observed and its location.
  • Skin Thickness The thickness of the skin was estimated related to the uninfected, untreated wound. This depth was estimated on the uniform histology photomicrographs from the surface to the muscle layer.
  • CONTROL GROUP 1 Uninfected and untreated.
  • Time Zero (See figures 14-15) At time zero following the wound stripping there was cellular covering of the surface. The dermal layer was not thickened. The hair follicles have a single cellular lining. There was minimal vascularity and no inflammation. The depth of the tissue was considered zero for future bench mark. 0+
  • CONTROL GROUP 2 Infected and untreated.
  • the experimental model provided evidence of a histological contrast between the control and experimental groups.
  • Control Group 2 that was wound stripped and infected showed a clear contrast to the uninfected Control Group 1 .
  • the skin stripped infected group there was loss of the epithelial cellular covering, no follicular cellular proliferation, marked increase in vascularity and little inflammatory response.
  • This histological condition provided clear contrast to the treatment groups.
  • All treatment groups by comparison showed healing response with multiple layer cellular proliferation on the surface, multiple layer cellular proliferation along the hair follicles, less vascularity, but an inflammatory cellular response in the dermis and muscular levels. See figures 14 - 31.
  • PCA at a concentration of 25 mM also showed collagen layer formation between the epidermis and dermis. (See figures/ photos 24 and 25). This response is beneficial in the use of anthocyanin and
  • anthocyanidins and metabolites thereof as a cosmetic agent to promote wound healing and improve skin health, including wrinkle reduction or removal.
  • This method of use of anthocyanin and anthocyanidin metabolites, and particularly PCA, is based upon the two fold response; the collagen layer increase and the skin swelling that increased the depth of the skin.
  • Example 7 PCA's effect on Pseudomonas Aeruginosa ATCC 700888 and
  • the inventor has shown that a composition comprising PCA was able to stop the growth of a biofilm formation as well as destroy already formed biofilms.
  • the biofilms tested were Pseudomonas Aeruginosa ATCC 700888 and Staphylococcus aureus ATCC 33591 (MRSA).
  • the bacteria (Pseudomonas Aeruginosa ATCC 700888 and
  • Staphylococcus aureus ATCC 33591 (MRSA) were placed in reactors and allowed to grow and form biofilms. Then cloths and metal were treated by coating with PCA solutions and then were left to dry. Two sets of the stainless steel mesh had crystals imbedded into the mesh to replicate placement into a mesh or coated joint implant.
  • a standard ASTM E-2647 drip flow biofilm reactor was used to grow a biofilm and the treated surfaces (as well as the control) were placed into the reactors and the biofilm was allowed to grow for about 6 hours. The samples received a continuous nutrient flow for an additional time period for about 48hours to promote a steady growth rate of the biofilm.
  • CFU Colony forming unites
  • Example 8 Spray on Solution of PCA and Time Study of PCA's effect on Pseudomonas Aeruginosa ATCC 700888 and Staphylococcus aureus ATCC 33591 (MRSA) biofilms
  • the invention also provides a method of blocking initial attachment of the bacteria to the implant and therefor preventing growth/development of a biofilm on an implant.
  • BALB/c mice were infected with Propionibacterium acnes via intradermal injection and treated topically with varying concentrations of a novel test compound, PCA, at 2, 24, 48 and 72 hours following challenge. Efficacy was evaluated by CFU analysis from skin samples harvested at 96 hours post challenge. [0246] These data demonstrate that P. acnes establishes a steady intradermal colonization in the skin of BALB/c mice. When administered topically, PCA at 60 mg/kg, demonstrated a bacteriostatic effect and reduced P. acnes CFU burden in mouse skin by a statistically significant amount. All lower amounts of PCA showed no such effect.
  • PCA a novel test compound
  • mice Female BALB/c mice, ordered from Harlan and weighing 17-19 g, were acclimated to housing conditions and handled in accordance with AUP number TP-18- 13. The animals were acclimated for 4 days prior to bacterial challenge. Only animals deemed healthy and fully immunocompetent were included in this study. Cages were prepared with 2 mice per cage.
  • Propionibacterium acnes (1 100; ATCC 6919), procured directly from the American Type Culture Collection.
  • each mouse was anesthetized in an Isoflurane induction chamber and the lesion site was cleared of hair. An area of approximately 2.0 cm x 2.0 cm of skin on the dorsal area of each mouse was cleared through use of the depilatory agent Nair®.
  • test treatment PCA
  • PCA test treatment
  • test article preparations were administered topically at 2, 24, 48 and 72 hours following the bacterial challenge. While untreated mice were harvested at 2 hours post infection, CFU burden was not detected. However at 96 hours post infection, the CFU burden rose from 6.0 Iog10 to 6.65 Iog10, indicating a successful inoculation.
  • mice were humanely euthanized and skin was aseptically removed from the infection site. Skin samples were placed in homogenation vials with 2.0 mL PBS, weighed and homogenized using a mini-bead beater. Homogenate was serially diluted and plated anaerobically on TSA agar plates for enumeration of colony forming units per gram of skin tissue. [0256] The mean bacterial burden of the untreated group at 96 hours was 6.65 Iog10. CFU levels in all treated groups were compared to the untreated group to determine statistical significance. Only the high dose of PCA (60 mg/kg) showed significant reduction of CFU burden when compared to the untreated control
  • Example 10 Antibiotic testing with PCA or 246 THBA using propylene glycol
  • PCA or 246 THBA were combined with propylene glycol (PPG).
  • PPG propylene glycol
  • the PPG was placed on a paper disc and then either PCA or 246 THBA was applied.
  • the paper disc was then placed on colonies of various bacterial in a Petri dish. At a certain uniform time they were inspected and classified in the following categories:
  • CZ clear zone of inhibition surrounding the sample and zone measured in millimeters (mm).
  • Example 11 PCA to sterilize/disinfect human skin study [0260] A randomized double blinded study was performed at Loma Linda Medical School. It involved 4 phases over two years' time. The methods were as follows:
  • Phase 1 The active test reagent was topically applied 1 .54% PCA in sterile water to the anterior shoulder region. This 1 .54% solution of PCA in water was used effectively in our prior animal wound studies.
  • the controls were Chloroprep (2% Chlorhexidine in 70% isopropyl alcohol) and Betadine (9.0% to 12.0% available iodine in water). Cultures were taken before application and 20 minutes after application. The initial harvest was by a surface swab. Application was by soaked sponge, without force or scrubbing. The second harvest was performed with the back edge of a sterile knife blade scraping with pressure in attempt to maximize the harvest from the deeper sebaceous glands and hair follicles. The specimens were placed in culture media.
  • Phase 2 included eleven medical students and was same method as Phase 1 . However the PCA vehicle was changed to 70% isopropyl alcohol. This allowed a higher concentration of PCA than possible in sterile water, 10%.
  • Phase 2a The 70% isopropyl alcohol vehicle was tested for its bactericidal properties. All cultures that were negative or markedly reduced with PCA topical solution were examined for exact nature of the index bacteria and the post treatment cultures that showed no or minimal growth. In this way it was learned what specific bacterial strains PCA could eliminate or reduce. Results:
  • Phase 1 showed the aqueous solution of 1 .54% PCA to be partially effective as compared to the controls.
  • 1 % PCA in water showed no growth in 7 of 22 subjects on aerobic culture and 10 cultures showed reduced growth.
  • 6 of 22 cultures showed no growth and 15 showed reduced colony growth.
  • Six heavy growth pre- treatment cultures prior to the 1.24% PCA treatment were chosen to examine the isolates to learn what pathogens were killed or not killed. Most of the bacteria eradicated were non pathogens.
  • Sample culture #29 showed 5 unique colonies by aerobic culture and 2 unique colonies by anaerobic culture. After 1 .24% PCA treatment there was no growth on either culture. Therefore pre-treatment cultures were examined for the species.
  • the chart below is the result of the specific species colony identified and the method of identification used. The organisms were predominately non pathogens except for P. acnes, all of which were removed by the treatment.
  • Propionibacterium acnes a potential pathogen.
  • the post treatment cultures showed the colonies of P. acnes was decreased to 5 colonies in one and 1 colony in another, but not eliminated. This suggested that PCA may be effective against P. acnes if a higher concentration was applied in subsequent Phases of this study. Testing with 1 .24% in water killed bacteria of normal flora as shown above chart. To increase efficacy and to improve skin penetration to the depth of the hair follicles that can harbor bacteria, Phase
  • Phase II This study involved 1 1 human subjects. There were 5 Males and 6 females. The ages were 23-33 years. There were two reagents. The control was 70% isopropyl alcohol.
  • the PCA source was a phytochemical extract from Nanjing Zelang Medical Technology Co. LTD. This source was chosen due to markedly reduced cost of goods compared to that which is biochemically manufactured.
  • the experimental dose was 9 +/- % PCA in 70% isopropyl alcohol. 10 grams of PCA was placed in 100 ml of isopropyl alcohol. The isopropyl alcohol allowed for a greater dose of PCA than water, (allowed more PCA to be dissolved).
  • the initial harvest was by a surface swab. Application was by soaked sponge, without force or scrubbing. The second harvest was performed with the back edge of a sterile knife blade scraping with pressure in attempt to maximize the harvest from the deeper sebaceous glands and hair follicles.
  • Phase 2 results with 9+% solution of PCA were compared to similar test Phase 1 and this showed this to comparable to Betadine in effectiveness, but not with Chloroprep, which killed all the bacterial colonies.
  • Phase 2a showed that 70% isopropyl alcohol (IPA) alone had few antibacterial properties. Of course sterile water used in Phase 1 had no antibacterial properties, therefore the testing was on the effectiveness of the PCA.
  • IPA isopropyl alcohol
  • PCA at 1.54 % although effective in an animal open skin wound was not a very effective antibacterial reagent on intact human skin. Therefore the concentration of PCA needed to be increased and could not be increased with water, but only be accomplished with the vehicle of 70% isopropyl alcohol.
  • Phase 3 Another test was conducted where the PCA was dissolved in 15 ml propylene glycol.
  • the PPH is a skin penetration enhancer and assists in dissolving PCA.
  • propylene glycol was added. Fasano WJ, ten Berge WF, Banton Ml, Heneweer M, Moore NP. Dermal penetration of propylene glycols: measured absorption across human abdominal skin in vitro and comparison with a QSAR model. Toxicol In Vitro. 201 1 Dec;25(8): 1664-70. Then to further the skin penetration an essential oil was added; i.e. an essence of peppermint oil (EOPO) (Nature oil, 1800 Miller Parkway, Streetsburo, OH 44241 100% pure [Japan]. Chen J, Jiang Q-D, Wu Y- M, Liu P, Yao J-H, Lu Q, Zhang H, Duan J-A. Potential of Essential Oils as Penetration Enhancers for Transdermal Administration of Ibuprofen to Treat Dysmenorrhoea
  • Staphylococcus epidermidis / hominis and Micrococcus luteus The Subject #18's post PCA treatment aerobic culture, #20 showed 2 colonies; S. capitis and S. epidermidis. For reasons unknown, neither of these species was identified in the index culture, and both are considered non pathogens.
  • the pre-PCA treatment index culture on subject #37 had colonies too numerous to count (TNTC) with heavy growth. The colony species were reported as one; Staphylococcus capitus.
  • the Subject #37's post PCA treatment aerobic culture #39 showed one (1 ) colony growth, but not the former S. capitus, but was identified Staphylococcus epidermidis as the residual. As in prior phases, the residual growth was most often a non pathogen on aerobic culture.
  • Propionibacterium avidium After this PCA treatment there was no anaerobic growth. Subject #37 index anaerobic culture showed 79 colonies. There were 4 prominent colony species; Staphylococcus epidermidis, Staphylococcus capitis /caprae,
  • Staphylococcus capitis and Propionibacterium acnes The one post PCA culture that showed a bacterial colony culture was subject #37 with post PCA treatment anaerobic culture being #39. There were 2 two colonies of same species; Staphylococcus epidermidis. There were no Propionibacterium acnes colonies. Thus, the results of Phase 3 showed the effectiveness of 17%+/- PCA in a composition of matter that had skin penetration properties; propylene glycol and essence of peppermint oil.
  • the solution used was 20 mg of ground PCA (Nanjing Zelang Medical Technology Co. Ltd.) in 95 ml 91 % isopropyl alcohol and 5 ml of essential oil of peppermint 100% pure .
  • the method placed the 20 grams of ground PCA in sterile container with volume markers. Then added 85 ml of 91 % isopropyl alcohol; warm and shake. 5 ml of essential oil of peppermint was added. A repeat of warming and shaking was instituted. The container was filled to 100 ml, warm and repeated the shaking. It took perhaps 15 to 20 minutes for the ground crystals to dissolve.
  • #8 pre PCA treatment anaerobic culture had 8 colony count but zero at 20 and 60 minutes. The interest was to learn what bacterial were eliminated. There were two species; S. capitis and S. epidermidis. #12 post PCA at 60 minutes showed 1 spreader colony on the plate (SPR) 50 colonies. This contrasted with the pre-treatment of 28 colonies. The species found in #12 aerobic were Klebsiella pneumonia/oxytoca and Micrococcus leuteus. These are not common pathogens. #14 pre PCA anaerobic showed 1 15 colonies. The species were Propionbacterium acnes, S. ludgunessis and Kocuia varians. There was no growth at 20 minutes showing that potential pathogen P. acnes was eliminated.
  • Gram Negative Rods Stenotrophomonas maltophilia: rare pathogen and Stenotrophomonas maltophilia.
  • the Gram Positive cocci were Kytococcus sedentarius: rare opportunistic pathogen and Micrococcus luteus / lylae: opportunistic pathogen, particularly in hosts with compromised immune systems, such as HIV patients.
  • #19 was a pre PCA anaerobic culture with 39 count showing species of P. acnes and S.
  • #32 was pre Chloroprep with aerobic colony count of 59 with the following species: Micrococcus luteus/lylae, Micrococcus luteus and Kocuria Kritinae. There was no colony growth at 20 or 60 minutes. #32 was pre Chloroprep anaerobic with 17 colonies with the following species: S. epidermidis and Gemella Bergeri/sanguinis. The post treatment showed no growth at 20 or 60 minutes. It should be noted in any of the literature or these experiments that re-colonization occurs with skin bacteria, likely in 30 minutes after treatment.
  • Species Analysis The pre-treatment species were predominately saprotrophic or commensal organisms. Propionibacterium acnes was identified in two subjects. After PCA treatment the specific species analysis showed predominately saprotrophic or commensal organisms. The potential pathogen Propionibacterium acnes was identified with 19 colonies pre-PCA treatment in #14 but showed no growth at 20 minutes after PCA treatment. P. acnes was identified pre-PCA treatment in #19 with 19 colonies and post treatment at 20 minutes there was one remaining colony of P. acnes. However there was no growth at 60 minutes.
  • PCA composition was not as effective as Cloroprep® against anaerobes, but where there was post PCA growth the species specific analysis showed that Propionibacterium was controlled.
  • PCA in one or more of these vehicles provided a broad spectrum disinfectant effect comparable to existing commercial products; isopropyl alcohol, Chloroprep® and Betadine®.
  • the other method is to establish clear superiority to an existing product. Both were considered in this study.
  • the various PCA solutions showed a non-inferiority status to 70% Isopropyl alcohol, Betadine® and Chloroprep®.
  • the PCA solutions in this study showed superiority to all but Chloroprep®.
  • the materials and methods were modified in attempt to improve the results of the test reagent PCA.
  • the vehicle was changed from water to isopropyl alcohol to facilitate an increase dose of PCA.
  • the vehicle was change to include reagents known to have skin penetration properties as well as antibiotic properties; i.e. propylene glycol and essence of peppermint oil.
  • Chloroprep® had a 7% incidence of residual bacteria; Propionibacterium acnes and Staphylococcus aureus persisted after treatment and his harvest was at a short time in which the reagent was still moist and there was not enough time perhaps for
  • Chloroprep® results in the literature since there were many variables in methods and timing of harvesting. There is not an apparent explanation for the differing results at 20 minutes in Phase I and IV in our study. Yet in Phase IV Chloroprep® was better than 20% PCA in 91 % isopropyl alcohol and 5 ml of essence of peppermint oil except at 20 minute aerobic cultures, which were sterile with the PCA solution. Note that isopropyl alcohol was not in Phase IV solution.
  • Statement 1 A method of reducing growth or development of a biofilm on a surface, including: contacting the surface with a composition including an effective amount of an anthocyanin metabolite or an anthocyanidin metabolite selected from protocatechuic acid and 2,4,6-trihydroxybenzaldehyde and mixtures and combinations thereof, wherein the contacting step reduces an initial attachment of a biofilm or a biofilm forming microorganism to the surface, thereby reducing the growth or
  • Statement 2 The method of statement 1 , wherein the biofilm forming microorganism is selected from a Pseudomonas aeruginosa and a Methicillin-resistant Staphylococcus aureus.
  • Statement 3 The method of statements 1 -2, wherein the composition is selected from a liquid, a sanitizer, a disinfectant, a swab, a salve, soap, foam, cream, solution, gel, spray, powder, crystals, antibacterial treatments, wipes, a lotion and combinations thereof.
  • Statement 4 The method of statements 1 -3, wherein the surface is selected from a solid surface, a smooth surface, a porous or semi-porous surface, and a cloth-like surface.
  • Statement 5 The method of statements 1 -4, wherein the surface is on an implant, a bandage, a catheter, a dressing, a heart or vascular grafts, or a wound.
  • Statement 6 The method of statements 1 -5, wherein the surface is a ceramic surface.
  • Statement 7 The method of statements 1 -6, wherein the contacting step is selected from wiping the surface, spraying the surface, soaking the surface, imbedding crystals of the anthocyanin metabolite or an anthocyanidin metabolite on or in the surface, or applying a bandage or wound dressing impregnated with the composition to the surface.
  • Statement 8 The method of statements 1 -7, wherein the anthocyanin metabolite or an anthocyanidin metabolite is present in an amount of from about 10% to about 50% by weight of the composition.
  • Statement 9 The method of statements 1 -8, wherein the anthocyanin metabolite or an anthocyanidin metabolite is present in an amount of from about 20% to about 30% by weight of the composition.
  • Statement 10 The method of statements 1 -9, wherein the anthocyanin metabolite or an anthocyanidin metabolite is protocatechuic acid.
  • Statement 1 1 The method of statements 1 -10, wherein the surface is contacted with the composition for up to two years.
  • Statement 12 The method of statements 1 -1 1 , wherein the composition comprises protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof dissolved in a solution.
  • Statement 13 The method of statements 3-12, wherein the solution further comprises a carrier.
  • Statement 14 The method of statement 13, wherein the carrier is selected from water, isopropyl alcohol, and ethyl alcohol.
  • Statement 15 The method of statement 14, wherein the isopropyl alcohol is present at a concentration of about 70% to about 91 %.
  • Statement 16 The method of statements 3-15, wherein the solution comprises 10% to 30% protocatechuic acid dissolved in about 70% isopropyl alcohol.
  • Statement 17 A method of destroying a biofilm on a surface, including: contacting the surface having one or more biofilms with a composition including an effective amount of an anthocyanin metabolite or an anthocyanidin metabolite selected from protocatechuic acid, 2,4,6-trihydroxybenzaldehyde, and mixtures and
  • biofilms are destroyed upon contact and one or more biofilm colonies surrounded by the biofilms are killed.
  • Statement 18 The method of statement 17, wherein the biofilm colony is selected from a Pseudomonas aeruginosa biofilm colony and a Methicillin-resistant Staphylococcus aureus biofilm colony.
  • Statement 19 The method of statements 17-18, wherein the surface is selected from a solid surface, a smooth surface, a porous or semi-porous surface, and a cloth-like surface.
  • Statement 20 The method of statements 17-19, wherein the surface is a ceramic surface.
  • Statement 21 The method of statements 17-20, wherein the contacting step is selected from wiping the surface, spraying the surface, soaking the surface, imbedding crystals of the anthocyanin metabolite or an anthocyanidin metabolite on or in the surface, or applying a bandage or wound dressing impregnated with the composition to the surface.
  • Statement 22 The method of statements 17-21 , wherein the surface is contacted with the composition for about 1 minute to about 60 minutes.
  • composition is a solution including about 10% to about 30% by weight of protocatechuic acid dissolved in about 70% isopropyl alcohol, whereby biofilms are destroyed upon contact and one or more biofilm colonies surrounded by the biofilms are killed and wherein the solution achieves a reduction in Pseudomonas aeruginosa or Methicillin- resistant Staphylococcus aureus colony forming units upon the surface.
  • Statement 24 The method of statements 17-23, wherein the contacting step comprises spraying the solution on the surface.
  • Statement 25 The method of statements 17-24, wherein greater than 99.9% of Pseudomonas aeruginosa biofilm colonies are killed after 30 minutes of being contacted with the composition.
  • Statement 26 The method of statements 17-25, wherein about 90% of Methicillin-resistant Staphylococcus aureus biofilm colonies are killed after 30 minutes of being contacted with the composition.
  • Statement 27 A bandage or wound dressing including protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof impregnated on the bandage or wound dressing.
  • Statement 28 The bandage or wound dressing of statement 27, wherein the protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof is part of a solution.
  • Statement 29 The bandage or wound dressing of statements 27-28, wherein the bandage or wound dressing has been sprayed or soaked with the solution, and wherein the protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof becomes activated by contact with moisture from the wound.
  • Statement 30 The bandage or wound dressing of statements 27-29, wherein the protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof
  • Statement 31 The bandage or wound dressing of statements 27-30, wherein the bandage or wound dressing is made of a material selected from the group consisting of a cloth material, linen, metal, and sintered metallic mesh.
  • Statement 32 The bandage or wound dressing of statements 27-31 , wherein the bandage or wound dressing is in the form of a patch.
  • Statement 33 The bandage or wound dressing of statements 27-32, wherein the protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or mixture or combination thereof is provided in an effective amount to promote an effect selected from the group consisting of inhibiting microorganism growth, eliminating microorganism growth, inhibiting microorganism survival, and eliminating microorganism survival.
  • Statement 34 The bandage or wound dressing of statements 27-33, wherein the bandage or wound dressing is impregnated with protocatechuic acid and not 2,4,6- trihydroxybenzaldehyde.
  • Statement 35 The bandage or wound dressing of statements 28-34, wherein the protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture thereof is present in the solution in a concentration of at least 25 mM.
  • Statement 36 The bandage or wound dressing of statements 28-35, wherein the protocatechuic acid is about 10% to about 40% by weight (w/v) of the solution.
  • Statement 37 The bandage or wound dressing of statements 28-36, wherein the solution further comprises a carrier that is selected from the group consisting of water, isopropyl alcohol, ethanol, propylene glycol, polyethylene glycol, or glycerol and mixtures or combinations thereof.
  • Statement 38 The bandage or wound dressing of statements 28-37, wherein the solution further comprises an essential oil.
  • Statement 39 The bandage or wound dressing of statements 28-38, wherein the solution further comprises isopropyl alcohol, propylene glycol, and an essential oil.
  • Statement 40 The bandage or wound dressing of statement 28-39, wherein the isopropyl alcohol is present as a solution including about 70% to about 91 % isopropyl alcohol.
  • Statement 41 The bandage or wound dressing of statement 33, wherein the microorganism is selected from the group consisting of C. difficile 9689, P. acnes 6919, C. Prefringens 13124, L. casei, E. coli 8739, E. coli 43895, S. Aureus 6538, S. Aureus 33591 , S. mutans, S. pyogenes, P. aeruginosa 9027, K. pneumonia, MRSE 51625, C. albicans, and legionella 43662.
  • the microorganism is selected from the group consisting of C. difficile 9689, P. acnes 6919, C. Prefringens 13124, L. casei, E. coli 8739, E. coli 43895, S. Aureus 6538, S. Aureus 33591 , S. mutans, S. pyogenes, P. aeruginosa 9027, K. pneumonia, MRSE
  • Statement 42 The bandage or wound dressing of statement 33, wherein the microorganism is part of a colony forming a biofilm.
  • Statement 43 A method of making a bandage or wound dressing impregnated with protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof including:
  • the effective amount of protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof elicits an effect selected from the group consisting of inhibiting microorganism growth, eliminating microorganism growth, inhibiting
  • Statement 44 The method of statement 43, wherein the bandage or wound dressing material is impregnated by soaking or spraying the bandage or wound dressing material with a solution including an alcohol selected from isopropyl alcohol and ethyl alcohol, and wherein the solution further comprises protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof, and wherein the protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof becomes activated by contact with moisture from the wound.
  • a solution including an alcohol selected from isopropyl alcohol and ethyl alcohol
  • the solution further comprises protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof, and wherein the protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof becomes activated by contact with moisture from the wound.
  • Statement 45 The method of statement 43, wherein the bandage or wound dressing material is impregnated with an effective amount of crystals of protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof.
  • Statement 46 A method for treating a patient having a wound including applying a bandage or wound dressing to the wound, wherein the bandage or wound dressing has been impregnated or will subsequently be impregnated with
  • the bandage or wound dressing elicits an effect selected from the group consisting of inhibiting microorganism growth, eliminating microorganism growth, inhibiting microorganism survival, and eliminating microorganism survival, thereby treating the wound.
  • Statement 48 The use or method of statements 46-47, wherein the bandage or wound dressing is first applied to the wound and the wound dressing is then soaked or sprayed with a solution including protocatechuic acid or 2,4,6- trihydroxybenzaldehyde or a mixture or combination thereof.
  • Statement 49 The use or method of statements 46-48, wherein the microorganism is selected from C. difficile 9689, P. acnes 6919, C. Prefringens 13124, L. casei, E. coli 8739, E. coli 43895, S. Aureus 6538, S. Aureus 33591 , S. mutans, S. pyogenes, P. aeruginosa 9027, K. pneumonia, MRSE 51625, C. albicans, and legionella 43662.
  • Statement 50 The use or method of statements 46-49, wherein the wound is a burn, skin break, bone break, muscle tear, puncture, surgical incision site, microdermabrasion site, skin graft site, a wound associated with diabetes, a bed sore, a pressure sore, skin defoliation, or a laceration, and wherein the protocatechuic acid or 2,4,6-trihydroxybenzaldehyde or a mixture or combination thereof becomes activated by contact with moisture from the wound.
  • Statement 51 A method of reducing growth of, development of, or destroying a biofilm on a surface, including: contacting the surface with a composition including an effective amount of an anthocyanin metabolite or an anthocyanidin metabolite selected from protocatechuic acid and 2,4,6-trihydroxybenzaldehyde and mixtures and combinations thereof, wherein the contacting step reduces an initial attachment of a biofilm or a biofilm forming microorganism to the surface, thereby reducing the growth or development of the biofilm on the surface.
  • Statement 52 The method of statement 51 , wherein the biofilm forming microorganism is selected from C. difficile 9689, P. acnes 6919, C. Prefringens 13124, L. casei, E. coli 8739, E. coli 43895, S. Aureus 6538, S. Aureus 33591 , S. mutans, S. pyogenes, P. aeruginosa 9027, K. pneumonia, MRSE 51625, C. albicans, and legionella 43662.
  • Statement 53 The method according to any one of statements 51 -52, wherein the biofilm forming microorganism is selected from a Pseudomonas aeruginosa and a Methicillin-resistant Staphylococcus aureus.
  • Statement 54 The method according to any one of statements 51 -53, wherein the composition is selected from a liquid, a sanitizer, a disinfectant, a swab, a salve, soap, foam, cream, solution, gel, spray, powder, crystals, antibacterial treatments, wipes, a lotion and combinations thereof.
  • Statement 55 The method according to any one of statements 51 -54, wherein the surface is selected from a solid surface, a smooth surface, a porous or semi-porous surface, and a cloth-like surface.
  • Statement 56 The method according to any one of statements 51 -55, wherein the surface is on an implant, a bandage, a catheter, a dressing, a heart or vascular grafts, or a wound.
  • Statement 57 The method according to any one of statements 51 -56, wherein the surface is a ceramic surface.
  • Statement 58 The method according to any one of statements 51 -57, wherein the contacting step is selected from wiping the surface, spraying the surface, soaking the surface, imbedding crystals of the anthocyanin metabolite or an
  • anthocyanidin metabolite on or in the surface or applying a bandage or wound dressing impregnated with the composition to the surface.
  • Statement 59 The method according to any one of statements 51 -58, wherein the anthocyanin metabolite or an anthocyanidin metabolite is present in an amount of from about 10% to about 50% by weight of the composition.
  • Statement 60 The method according to any one of statements 51 -59, wherein the anthocyanin metabolite or an anthocyanidin metabolite is present in an amount of from about 20% to about 30% by weight of the composition.
  • Statement 61 The method according to any one of statements 51 -60, wherein the anthocyanin metabolite or an anthocyanidin metabolite is protocatechuic acid.
  • Statement 62 The method according to any one of statements 51 -61 , wherein the surface is contacted with the composition for up to two years.
  • Statement 63 The method according to any one of statements 51 -62, wherein the composition comprises protocatechuic acid or 2,4,6- trihydroxybenzaldehyde or a mixture or combination thereof dissolved in a solution.
  • Statement 64 The method according to any one of statements 54-63, wherein the solution further comprises a carrier.
  • Statement 65 The method according to statement 64, wherein the carrier is selected from water, isopropyl alcohol, and ethyl alcohol.
  • Statement 66 The method according to statement 65, wherein the isopropyl alcohol is present at a concentration of about 70% to about 91 %.
  • Statement 67 The method according to any one of statements 54-66, wherein the solution comprises 10% to 30% protocatechuic acid dissolved in about 70% isopropyl alcohol.
  • Statement 68 The method according to any one of statements 51 -67, wherein the surface is contacted with the composition for about 1 minute to about 60 minutes.
  • Statement 69 The method according to any one of statements 51 -68, wherein greater than 99.9% of Pseudomonas aeruginosa biofilm colonies are killed after 30 minutes of being contacted with the composition.
  • Statement 70 The method according to any one of statements 51 -69, wherein about 90% of Methicillin-resistant Staphylococcus aureus biofilm colonies are killed after 30 minutes of being contacted with the composition.
  • Statement 71 The method according to any one of statements 51 -70, wherein the surface is a bandage or wound dressing and the contacting step further comprises impregnating the composition including the anthocyanin metabolite or an anthocyanidin metabolite on the bandage or wound dressing.
  • Statement 72 The method according to statement 71 , wherein the anthocyanin metabolite or an anthocyanidin metabolite is a crystal.
  • Statement 73 The method according to any one of statements 71 -72, wherein the anthocyanin metabolite or an anthocyanidin metabolite is part of a solution.
  • Statement 74 The method according to any one of statements 71 -73, wherein the contacting step comprises spraying or soaking the bandage or wound dressing with the anthocyanin metabolite or an anthocyanidin metabolite and drying the bandage or wound dressing, wherein the anthocyanin metabolite or an anthocyanidin metabolite becomes activated by contact with moisture from a wound.
  • Statement 75 The method according to any one of statements 71 -74, wherein the bandage or wound dressing is made of a material selected from the group consisting of a cloth material, linen, metal, and sintered metallic mesh.
  • Statement 76 The method according to any one of statements 71 -75, wherein the bandage or wound dressing in the form of a patch.
  • Statement 77 The method according to any one of statements 71 -76, wherein the anthocyanin metabolite or an anthocyanidin metabolite is protocatechuic acid and not 2,4,6- trihydroxybenzaldehyde.
  • Statement 78 A bandage or wound dressing prepared by the method according to any one of statements 71 -77.

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Abstract

La présente invention concerne d'une manière générale des compositions antimicrobiennes et des procédés d'utilisation comprenant une anthocyanine, une anthocyanidine ou leurs métabolites. La présente invention concerne également des procédés destinés à favoriser la cicatrisation d'une plaie en utilisant ces compositions. Ces compositions présentent une activité antimicrobienne à large spectre et sont sûres pour les utilisations humaines et animales. En outre, ces compositions sont sûres pour les utilisations médicales et les utilisations industrielles comme préparations antiseptiques afin de réduire ou d'empêcher la croissance microbienne, comprenant la destruction des biofilms bactériens.
PCT/US2018/027157 2017-04-11 2018-04-11 Agents antimicrobiens et leurs procédés d'utilisation Ceased WO2018191409A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020033677A1 (fr) * 2018-08-08 2020-02-13 Johnson Lanny Leo Méthodes de diagnostic et de traitement d'implants infectés
US10772860B2 (en) 2013-05-01 2020-09-15 Lanny Leo Johnson Antimicrobials and methods of use thereof
EP4052575A1 (fr) * 2021-03-03 2022-09-07 Berkem Developpement Utilisation d'un extrait de raisin en tant que virucide a l'encontre des virus de la famille des coronavirus
KR20240086745A (ko) * 2022-11-29 2024-06-19 충남대학교산학협력단 염화 시아니딘을 유효성분으로 포함하는 생물막 형성 억제용 조성물

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140328902A1 (en) * 2013-05-01 2014-11-06 Lanny Leo Johnson Antimicrobials and methods of use thereof for wound healing
CN105647674A (zh) * 2016-04-06 2016-06-08 苏州甫众塑胶有限公司 一种高效卫生管道清洗剂及其制备方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140328902A1 (en) * 2013-05-01 2014-11-06 Lanny Leo Johnson Antimicrobials and methods of use thereof for wound healing
CN105647674A (zh) * 2016-04-06 2016-06-08 苏州甫众塑胶有限公司 一种高效卫生管道清洗剂及其制备方法

Non-Patent Citations (47)

* Cited by examiner, † Cited by third party
Title
"CDC, Vital Signs; Preventing Clostridium difficile infections", MMWR, 2012, pages 61 - 157,62
A. MORÁN ET AL: "Non-toxic plant metabolites regulate Staphylococcus viability and biofilm formation: a natural therapeutic strategy useful in the treatment and prevention of skin infections", BIOFOULING: THE JOURNAL OF BIOADHESION AND BIOFILM RESEARCH, vol. 30, no. 10, 14 November 2014 (2014-11-14), GN, pages 1175 - 1182, XP055487430, ISSN: 0892-7014, DOI: 10.1080/08927014.2014.976207 *
ANDERSON, JB, NAT REV MICROBIOL, vol. 3, no. 7, 2005, pages 547
ANGLEN J: "Perspectives on Modern Orthopedics. Wound Irrigation in Musculoskeletal Injury", J AM ACAD ORTHO SURG., vol. 9, no. 4, July 2001 (2001-07-01), pages 219 - 226
BACHMANN, SP, ANTIMICROBIAL AGENTS CHEMOTHER, vol. 46, 2002, pages 3591
BAKHSHESHIAN J; DAHDALEH NS; LAM SK; SAVAGE JW; SMITH ZA: "The use of vancomycin powder in modern spine surgery: systematic review and meta-analysis of the clinical evidence", WORLD NEUROSURG, vol. 83, no. 5, May 2015 (2015-05-01), pages 816 - 23
BLASER, M.: "Antibiotic overuse: Stop the killing of beneficial bacteria", NATURE, vol. 476, 25 August 2011 (2011-08-25), pages 393 - 394
BOZIC KJ ET AL.: "The epidemiology of revision total knee arthroplasty in the United States", CLIN ORTHOP RELAT RES., vol. 468, no. 1, January 2010 (2010-01-01), pages 45 - 51
BUSUIOC CJ ET AL.: "Phases of cutaneous angiogenesis process in experimental third-degree skin burns: histological and immunohistochemical study", ROM J MORPHOL EMBRYOL, vol. 54, no. 1, 2013, pages 163 - 710
CHARACKLIS WG; MCFETERS GA; MARSHALL KC: "Physiological ecology in biofilm systems", 1990, JOHN WILEY & SONS, pages: 341 - 94
CHAWEEWANNAKOM U. ET AL.: "Cost analysis of peri-operative antibiotic administration in total knee arthroplasty", J MED ASSOC THAI., vol. 95, no. 10, October 2012 (2012-10-01), pages S42 - 7
CHEN J; JIANG Q-D; WU Y-M; LIU P; YAO J-H; LU Q; ZHANG H; DUAN J-A: "Potential of Essential Oils as Penetration Enhancers for Transdermal Administration of Ibuprofen to Treat Dysmenorrhoea", MOLECULES, vol. 20, 2015, pages 18219 - 18236
CRONQUIST AB; JOKOB K; LAI L; LATTA PD; LARSON EL: "Relationship between Skin Microbial Counts and Surgical Site Infection after Neurosurgery", CLIN INFECT DIS., vol. 33, no. 8, 2001, pages 1302 - 1308
D'AGOSTINO SR RB; MASSAROL JM; SULLIVAN LM: "Non-inferiority trials: design concepts and issues - the encounters of academic consultants in statistics", STATIST. MED., vol. 22, 2003, pages 169 - 186
DATABASE WPI Week 201654, 8 June 2016 Derwent World Patents Index; AN 2016-39115S, XP002782466 *
DE VITA DANIELA ET AL: "Activity of caffeic acid derivatives againstCandida albicansbiofilm", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, vol. 24, no. 6, 13 February 2014 (2014-02-13), pages 1502 - 1505, XP028625290, ISSN: 0960-894X, DOI: 10.1016/J.BMCL.2014.02.005 *
DONLAN R M: "Biofilms: Microbial life on surfaces", EMERGING INFECTIOUS DISEASES, EID, ATLANTA, GA, US, vol. 8, no. 9, 1 September 2002 (2002-09-01), pages 881 - 890, XP002384848, ISSN: 1080-6040 *
DONLAN RM: "Biofilms: Microbial Life on Surfaces", EMERG INFECT DISEASE, vol. 8, no. 9, 2002, pages 881 - 890, XP002384848
DONLAN RM: "Role of biofilms in antimicrobial resistance", ASAIO J., vol. 46, 2000, pages 47 - 52
DONLAN, RM: "Biofilms: Microbial Life on Surfaces", EMERGING INFECTIOUS DISEASES, vol. 8, no. 9, September 2002 (2002-09-01), XP002384848
E TAMPAKAKIS; A Y PELEG; E MYLONAKIS, EUKARYOT CELL, vol. 8, 2009, pages 732
FASANO WJ; TEN BERGE WF; BANTON MI; HENEWEER M; MOORE NP: "Dermal penetration of propylene glycols: measured absorption across human abdominal skin in vitro and comparison with a QSAR model", TOXICOL IN VITRO, vol. 25, no. 8, December 2011 (2011-12-01), pages 1664 - 70
FUX, CA, TRENDS MICROBIOL, vol. 13, no. 1, 2005, pages 34
GARG P ET AL.: "Antibiotic-impregnated articulating cement spacer for infected total knee arthroplasty", INDIAN J ORTHOP., vol. 45, no. 6, November 2011 (2011-11-01), pages 535 - 540
GHARZI A; REYNOLDS AJ; JAHODA CA: "Plasticity of hair follicle dermal cells in wound healing and induction", EXP DERMATOL., vol. 12, no. 2, April 2003 (2003-04-01), pages 126 - 36, XP002326459, DOI: doi:10.1034/j.1600-0625.2003.00106.x
GUTIÉRREZ SERGIO ET AL: "The Usefulness of Non-Toxic Plant Metabolites in the Control of Bacterial Proliferation", PROBIOTICS AND ANTIMICROBIAL PROTEINS, NEW YORK, NY ; HEIDELBERG : SPRINGER, NEW YORK, NY ; HEIDELBERG : SPRINGER, vol. 9, no. 3, 29 March 2017 (2017-03-29), pages 323 - 333, XP036297096, ISSN: 1867-1306, [retrieved on 20170329], DOI: 10.1007/S12602-017-9259-9 *
HAENLE M. ET AL.: "Economic impact of infected total knee arthroplasty", SCIENTIFIC WORLD JOURNAL, vol. 2012, 2012, pages 1 96515
HEBERT CK ET AL.: "Cost of treating an infected total knee replacement", CLIN ORTHOP RELAT RES., vol. 331, October 1996 (1996-10-01), pages 140 - 5
HELLER S; REZAPOOR M; PARVIZI J: "Minimising the risk of infection: a peri-operative checklist", BONE JOINT J., vol. 98-B, no. 1, January 2016 (2016-01-01), pages 18 - 22
J. UBEROS ET AL: "Phenolic acid content and antiadherence activity in the urine of patients treated with cranberry syrup (Vaccinium macrocarpon) vs. trimethoprim for recurrent urinary tract infection", JOURNAL OF FUNCTIONAL FOODS, vol. 18, 1 October 2015 (2015-10-01), NL, pages 608 - 616, XP055487411, ISSN: 1756-4646, DOI: 10.1016/j.jff.2015.08.009 *
JOHADA CA; REYNOLDS AJ: "Hair follicle dermal sheath cells: unsung participants in wound healing", LANCET, vol. 358, no. 9291, 27 October 2001 (2001-10-27), pages 1445 - 8
KAPADIA BH ET AL.: "The Economic Impact of Periprosthetic Infections Following Total Knee Arthroplasty at a Specialized Tertiary-Care Center", J ARTHROPLASTY, 17 October 2013 (2013-10-17)
KURTZ SM ET AL.: "Economic burden of periprosthetic joint infection in the United States", J ARTHROPLASTY, vol. 27, no. 8, September 2012 (2012-09-01)
LEE MJ; POTTINGER PS; BUTLER-WU S; BUMGARNE RE; RUSS SM; MATTSENLLL FA: "Propionibacterium Persists in the Skin Despite Standard Surgical Preparation", J BONE JOINT SURG AM, vol. 96, no. 17, 3 September 2014 (2014-09-03), pages 1447 - 1450
MCLELLAN E; ROWNSEND R; PARSONS HK: "Evaluation of ChloraPrep (2% chlorhexidine gluconate in 70% isopropyl alcohol) for skin antiseptic in preparation for blood culture collection", JOURNAL OF INFECTION, vol. 57, no. 6, pages 459 - 463
MELISSA JJ ET AL., ANTIMICROB AGENTS CHEMOTHER., vol. 53, no. 6, 2009, pages 2638
MOLINARI RW; KHERA OA; MOLINARI III WJ: "Prophylactic intraoperative powdered vancomycin and postoperative deep spinal wound infection: 1,512 consecutive surgical cases over a 6-year period", EUR SPINE J., vol. 21, no. 4, June 2012 (2012-06-01), pages 476 - 482, XP035067695, DOI: doi:10.1007/s00586-011-2104-z
MURGA R; MCALLISTER S; MILLER JM; TENOVER F; BELL M; DONLAN RM: "Effect of vancomycin treatment of methicillin-resistant S. aureus (MRSA) biofilms on central venous catheters in a model system", 2001 AMERICAN SOCIETY FOR MICROBIOLOGY ANNUAL MEETING, 23 May 2001 (2001-05-23)
NAKATSUJI T; CHIANG HI; JIANG SB; NAGARAJAN H; ZENGLER K; GALLO RL: "The microbiome extends to subepidermal compartments of normal skin", NAT COMMUN, vol. 4, 2013, pages 1431
ROMANO A; CAUBET JC: "Antibiotic allergies in children and adults: from clinical symptoms to skin testing diagnosis", J ALLERGY CLIN IMMUNOL PRACT., vol. 2, no. 1, January 2014 (2014-01-01), pages 3 - 12
SALTZMAN MD; NUBER GW; GRYZLO SM; MARECK GS; KOH JL: "Efficacy of Surgical Preparation Solutions in Shoulder Surgery", J BONE JOINT SURG AM, vol. 91, no. 8, 1 August 2009 (2009-08-01), pages 1949 - 1953
SEDOR J; MULHOLLAND SG: "Hospital acquired urinary tract infections associated with the indwelling catheter", UROL CLIN NORTH AM., vol. 26, 1999, pages 821 - 8
SEHULSTER L: "Centers for Disease Control and Prevention, Guidelines for environmental infection control in healthcare facilities", MMWR, vol. 52, no. RR10, 2003, pages 1 - 42
TAMPAKAKIS, E., EUKARYOT CELL, vol. 8, 2009, pages 732
WARREN, NG, AMERICAN SOCIETY FOR MICROBIOLOGY, 1995, pages 723
WHITESIDE LA: "Prophylactic peri-operative local antibiotic irrigation", BONE JOINT J., vol. 98-B, no. 1, January 2016 (2016-01-01), pages 23 - 6
ZEEUWEN PL; BOEKHORST J; VAN DEN BOGAARD EH; DE KONING HD; VAN DE KERKHOF PM; SAULNIER DM; VAN SWAM II; VAN HIJUM SA; KLEEREBEZEM: "Microbiome dynamics of human epidermis following skin barrier disruption", GENOME BIOL., vol. 13, no. 11, 15 November 2012 (2012-11-15), pages 101, XP021138391, DOI: doi:10.1186/gb-2012-13-11-r101

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WO2020033677A1 (fr) * 2018-08-08 2020-02-13 Johnson Lanny Leo Méthodes de diagnostic et de traitement d'implants infectés
EP4052575A1 (fr) * 2021-03-03 2022-09-07 Berkem Developpement Utilisation d'un extrait de raisin en tant que virucide a l'encontre des virus de la famille des coronavirus
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