Drug – bio-affecting and body treating compositions – Preparations characterized by special physical form – Biocides; animal or insect repellents or attractants
Reexamination Certificate
1998-08-28
2001-09-11
Page, Thurman K. (Department: 1615)
Drug, bio-affecting and body treating compositions
Preparations characterized by special physical form
Biocides; animal or insect repellents or attractants
C510S276000, C510S300000, C510S360000, C510S530000
Reexamination Certificate
active
06287585
ABSTRACT:
The present invention relates to a method of killing or inhibiting microbial cells or microorganisms, more specifically microbial cells or microorganisms present in laundry, on hard surface, on skin, teeth or mucous membranes; and for preserving cosmetics etc.
BACKGROUND OF THE INVENTION
At this time of increased public interest in reducing the use of chemical additives, it is relevant to consider natural alternatives for antimicrobial agents used e.g. for preserving foods and cosmetics, as disinfectants, and as an antimicrobial ingredient of detergent and cleaning compositions. This has increased interest in preservation using live bacterial cultures (Jeppesen & Huss 1993) and enzymes like lactoperoxidase (Farrag & Marth 1992), glucose oxidase (Jeong et al. 1992) and lysozyme (Johansen et al. 1994).
Gram-negative bacteria are often resistant to a large number of harmful agents due to the effective permeability barrier function of the outer membrane (Nakae 1985). However, certain cationic peptides or polymers are under certain conditions apparently able to traverse the outer membrane of Gram-negative bacteria (Vaara 1992, Vaara & Vaara 1983), probably as a result of their binding to the anionic lipopolysaccharide-covered surface of the Gram-negative cell. The mechanism of the antibacterial action of basic peptides is not known, but it has been suggested that small or short cationic polymers can form a channel in the cytoplasmic membrane, thus uncoupling electron transport and causing leakage (Christensen et al. 1988; Hugo 1978; Kagan et al. 1990). It has also been proposed that they induce autolysis due to activation of the autolytic enzymes (Bierbaum & Sahl 1991). The larger or longer cationic basic polymers agglutinate the cells and may thereby inhibit growth.
Thus, the object of invention is co provide a method of killing or inhibiting microbial cells which is easy to use, relatively inexpensive, and more effective than the known disinfecting preserving methods.
SUMMARY OF THE INVENTION
Surprisingly, it has been found that the combined action of an enzyme and a cationic polymer, when applied to e.g. a hard surface, skin, mucous membranes or laundry, results in a hitherto unknown synergistic antimicrobial effect.
Thus, based on these findings the present invention provides, in a first aspect, a method of inhibiting microorganisms present in laundry, wherein the laundry is treated with a soaking, washing or rinsing liquor comprising a poly-cationic compound and one or more enzymes, with the proviso that when the enzyme is an oxidoreductase then the poly-cationic compound is not a polylysine, polyarginine or a co-polymer thereof.
In a second aspect, the present invention provides a method of inhibiting microbial growth on a hard surface, wherein the surface is contacted with a composition comprising a poly-cationic compound and one or more enzymes, with the proviso that when the enzyme is an oxidoreductase then the poly-cationic compound is not a polylysine, polyarginine or a co-polymer thereof.
In a third aspect, the present invention provides a method of killing microbial cells present on human or animal skin, mucous membranes, teeth, wounds, bruises or in the eye or inhibiting the growth thereof, wherein the cells to be killed or inhibited or the skin, mucous membrane, teeth, wound or bruise are/is contacted with a composition comprising a poly-cationic compound and one or more enzymes, with the proviso that when the enzyme is an oxidoreductase then the poly-cationic compound is not a polylysine, polyarginine or a co-polymer thereof.
The composition used in the methods of the invention is useful as antimicrobial ingredient wherever such an ingredient is needed, for example for the preservation of cosmetics, contact lens products, or enzyme compositions; as a disinfectant for use e.g. on human or animal skin, mucous membranes, wounds, bruises or in the eye; for killing microbial cells in laundry; and for incorporation in cleaning compositions or disinfectants for hard surface cleaning or disinfection.
DETAILED DESCRIPTION OF THE INVENTION
In the present context, the term “bactericidal” is to be understood as capable of killing bacterial cells.
In the present context, the term “bacteriostatic” is to be understood as capable of inhibiting bacterial growth, i.e. inhibiting growing bacterial cells.
In the present context, the term “fungicidal” is to be understood as capable of killing fungal cells.
In the present context, the term “fungistatic” is to be understood as capable of inhibiting fungal growth, i.e. inhibiting growing fungal cells.
The term “growing cell” is to be understood as a cell having access to a suitable nutrient and thus being capable of reproduction/propagation. By the term “non-growing cell” is meant a living, but dormant, cell, i.e. a cell in the non-growing, non-dividing, non-multiplying and non-energized state with metabolic processes at a minimum.
The term “microbial cells” denotes bacterial or fungal cells, and the term “microorganism” denotes a fungus, a bacterium and a yeast.
The term “hard surface” as used herein relates to any surface which is essentially non-permeable for microorganisms. Examples of hard surfaces are surfaces made from metal, plastics, rubber, board, glass, wood, paper, textile, concrete, rock, marble, gypsum and ceramic materials which optionally may be coated, e.g. with paint, enamel and the like.
The Enzyme
The term “oxidoreductase” means an enzyme classified as EC 1. according to the Enzyme Nomenclature (1992), i.e. any enzyme classified as EC 1.1 (acting on the CH—OH group of donors), EC 1.2 (acting on the aldehyde or oxo group of donors), EC 1.3 (acting on the CH—CH group of donors), EC 1.4 (acting on the CH—NH
2
group of donors), EC 1.5 (acting on the CH—NH group of donors), EC 1.6 (acting on NADH or NADPH), EC 1.7 (acting on other nitrogenous compounds as donors), EC 1.8 (acting on a sulfur group of donors), EC 1.9 (acting on a heme group of donors), EC 1.10 (acting on diphenols and related substances as donors), EC 1.11 (acting on a peroxide as acceptor), EC 1.12 (acting on hydrogen as donor), EC 1.13 (acting on single donors with incorporation of molecular oxygen (oxygenases), EQ 1.14 (acting on paired donors with incorporation of molecular oxygen), EC 1.15 (acting on superoxide radicals as acceptor), EC 1.16 (oxidizing metal ions), EC 1.17 (acting on —CH
2
— groups), EC 1.18 (acting on reduced ferredoxin as donor), EC 1.19 (acting on reduced flavodoxin as donor), and EC 1.97 (other oxidoreductases).
The term “peroxidase enzyme system” is to be understood as a peroxidase (EC 1.11.1) in combination with a source of hydrogen peroxide which may be hydrogen peroxide or a hydrogen peroxide precursor for in situ production of hydrogen peroxide, e.g. percarbonate or perborate, or a hydrogen peroxide generating enzyme system, e.g. an oxidase and a substrate for the oxidase or an amino acid oxidase and a suitable amino acid, or a peroxycarboxylic acid or a salt thereof.
Examples of useful peroxidases are lactoperoxidase, horseradish peroxidase, peroxidases producible by cultivation of a peroxidase producing strain
Myxococcus virescens,
DSM 8593,
Myxococcus fulvus,
DSM 8969, or
Myxococcus xanthus,
DSM 8970, of a peroxidase producing strain of the genus Corallococcus, preferably belonging to
Corallococcus coralloides,
DSM 8967, or
Corallococcus exiguus,
DSM 8969.
Examples of useful substrates for peroxidase are thiocyanate, iodide, phenothiazins, syringates.
Laccases are enzymes that catalyze the oxidation of a substrate with oxygen, they are known from microbial, plant and animal origins. More specifically, laccases (EC 1.10.3.2) are oxidoreductases that function with molecular oxygen as electron acceptor. Molecular oxygen from the atmosphere will usually be present in sufficient quantity, so normally it is not necessary to add extra oxygen to the process medium. Examples of a laccase enzyme useful in the compositions of the present invention is laccase obtainable from the strain
Coprinus cinereus,
IFO 30116, or from a
Garbell John I.
Lambiris Elias J.
Novozymes A/S
Page Thurman K.
Seidleck Brian K.
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