JP2010202563A - Isochromene-based antimicrobial agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an antimicrobial agent safe to human body and high in antimicrobial activity on genus Legionella bacteria. <P>SOLUTION: The antimicrobial agent against genus Legionella bacteria is provided, including as an active ingredient, an isochromene derivative represented by formula (I) (wherein, R<SP>1</SP>is H, 2-pentyl, or 4,6-hydroxyphenyl); wherein the isochromene derivative may be extracted from Oakmoss or Evernia prunastri(L.) Arch.in Latin, which is a moss adhering on a skin of an oak tree etc. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an antibacterial agent against Legionella spp. Containing an isochromene derivative as an active ingredient.

  Legionella is a glucose non-fermented gram-negative bacillus widely distributed in nature such as water and soil, and causes bacterial infections with pneumonia as the main feature. This bacterium is an intracellular proliferating bacterium, and can grow in a living body without being sterilized in phagocytic cells such as macrophages. Moreover, since it is a β-lactamase-producing bacterium, it exhibits resistance to some β-lactam agents.

Many Legionella infections are caused by bacteria that have entered the foreign body. Examples of infection sources include cooling water for air conditioning, hot spring water, and a puddle of aging water supply facilities. In recent years, Legionnaires disease (Legionnaires disease) problems due to Legionella spp. That survive in open circulation system cooling water such as air conditioning equipment have frequently occurred. When these cooling towers are close to the window of the living room, the outside air intake of the air conditioner, etc., the aerosol of the cooling water enters the room and causes legionellosis.
Especially during the summer season when air conditioning is necessary, the water temperature of the cooling tower becomes high (about 15 ° C. to 35 ° C.), making it a suitable place for the growth of microorganisms such as Legionella and pathogens.

More recently, it is a "24-hour bath" (a device that has the function of purifying dirty hot water by keeping the bath water at an appropriate temperature 24 hours a day, eliminating the need for water replacement and daily cleaning). The growth of Legionella is regarded as a problem.
Historically, in 1976, when a US National Military Congress was held at a hotel in Philadelphia, USA, serious pneumonia of unknown cause occurred in a mass, and 29 of them died. The fungus (Legionella pneumophila) was found to be the causative bacterium.

  In Japan, since the first case of Legionella pneumonia was reported in 1981, many reports such as nosocomial infections in hospitals have been made. As a countermeasure against such problems caused by microorganisms, a method for suppressing the growth of microorganisms and bacteria by injecting an antibacterial agent into a circulating water system, etc., cleaning the inside of the apparatus physically, or chemically using a cleaning agent For example, a cleaning method has been used.

Antibiotics such as erythromycin and tafampicin are said to be effective against Legionella spp., But daily use of these antibiotics is not preferred in terms of appearance of resistant bacteria and side effects. In addition, Legionella pneumonia has a problem in that it progresses quickly and has a profound effect on the prognosis of life, so that prompt definitive diagnosis is not easy although it requires immediate treatment.
Therefore, it is said that prevention of contamination of the cooling water of the water-cooled air conditioner, which is the main infection source, is the most effective means as a countermeasure against legionellosis.

In order to sterilize infection sources such as cooling water for air conditioning, antibacterial agents can be injected to suppress the growth of bacteria. Halogen-based antibacterial agents such as chlorine, bromine and chlorine dioxide, and quaternary ammonium salts Organic compounds such as surfactants, hydrazines and maleimides, isothiazoline compounds, phosphonium compounds and nitroalcohol compounds are mainly used.
However, the halogen compound has a problem that it easily corrodes equipment and piping. On the other hand, the organic compound has a problem of toxicity such as skin irritation, sensitization, and carcinogenicity and insufficient efficacy. .

Under such circumstances, the idea that the prevention of Legionella infection is important is spreading, and there is a strong demand for providing an antibacterial active material that can be used easily by ordinary people and has no safety problems.
Under these circumstances, a method for controlling Legionellaceae gram-negative bacteria in an aqueous system by adding a seed extract of citrus fruits such as grapefruit has been proposed (Patent Document 1).

  In addition, Enmaiso, Oobanjutsu, Karakoboku, Wakouboku, Kankyo, Goldfish, Keigai, Genka (Sweet Flower), Suzuzoku (Grass Bean), Takran, Bangka (Sweet Fruit), Mitsumoka (Nectarflower), Yakuchi, Ryokyo, Guava leaf, Goryo leaf, Nanten leaf, Eucalyptus leaf, Artemisia, Guava fruit, Stevia, Mangosteen peel, Mulberry bark, Catnip, Cardamom, Sweet violet, Tarragon, Chives, Hyssop, Blackberry, Magwort, Monalda, Yarrow, Raspberry An anti-Legionella composition comprising one or more extracts selected from rosemary, wild strawberry or propolis extracts as an active ingredient has also been proposed (Patent Document 2). However, the effect was not high enough to completely suppress the growth of Legionella spp.

  Furthermore, the present invention shows that an extract of Oakmoss (Latin name Evernia prunastri (L.) Arch.), Which is a moss that grows on the epidermis of birch trees, exhibits antibacterial activity against Legionella. They have found (Non-Patent Document 1). However, the effect requires a concentration of 50 to 150 ppm to completely suppress the growth of Legionella spp., And the oak moss extract itself has a dark brown color and has a strong scent and is practically used. There was a problem with generalization.

JP-A-6-233986 Japanese Patent Laid-Open No. 11-43442

The 55th Annual Meeting of the Japanese Biopharmaceutical Society (2008), 268

  The problem to be solved by the present invention is to provide an anti-Legionella agent that is safe without adversely affecting the human body even if it is used on a daily basis, and that can potently inhibit the growth of Legionella genus bacteria. is there.

  Therefore, the present inventors have continued research on ingredients having antibacterial activity from natural products, and as part of that, attention has been focused on oak moss extract as a perfume raw material that has been used in high-end brand perfumes. As a result, they have intensively researched and found that only a specific component in the oak moss extract has a strong Legionella growth inhibitory action, thereby completing the present invention.

〔式中、Ｒ¹は水素原子または下記式（II）：

（式中Ｒ²は水素原子または炭素数１〜６の直鎖または分枝鎖状アルキル基である）の基を示す〕
で表されるイソクロメン誘導体を有効成分として含有することを特徴とするレジオネラ属菌に対する抗菌剤である。 That is, the present invention is as follows.
The following formula (I):

[Wherein R ¹ is a hydrogen atom or the following formula (II):

(Wherein R ² represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms)]
It is an antibacterial agent with respect to Legionella genus bacteria characterized by including the isochromene derivative represented by these as an active ingredient.

の化合物、または下記式（２）：

の化合物、または下記式（３）：

の化合物であることを特徴とする。
また、本発明は、上記の抗菌剤を含有することを特徴とする浴用剤、工業用殺菌剤、並びに浴槽洗浄剤である。 And the isochromene derivative has the following formula (1):

Or a compound of the following formula (2):

Or a compound of the following formula (3):

It is characterized by being said compound.
Moreover, this invention is a bath agent, an industrial disinfectant, and a bathtub cleaning agent characterized by containing said antibacterial agent.

The antibacterial agent of the present invention is derived from an oak moss extract that has been widely used as a cosmetic fragrance so far, and is safe with no side effects on the human body even when used on a daily basis. It is an anti-legionella spp.
In addition, because it does not have the unique smell of seaweed and moss like oak moss, and its color is almost colorless and transparent, it can be widely applied to daily necessities such as bathing agents and industrial disinfectants that are frequently used on a daily basis. is there.
Therefore, by using the antibacterial agent of the present invention, it is possible to control Legionella spp. Surviving in cooling / warming water systems such as circulating cooling water for cooling / refrigeration / refrigeration equipment and circulating hot water for 24-hour baths and heat storage water systems. , Can prevent its growth.

〔式中、Ｒ¹は水素原子または下記式（II）：

の基を示す〕
で表される化合物である。 [1] Isochromene derivative The isochromene derivative which is an active ingredient of the antibacterial agent of the present invention is
The following formula (I):

[Wherein R ¹ is a hydrogen atom or the following formula (II):

Shows the group of
It is a compound represented by these.

In the formula (II), R ² is a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms, preferably hydrogen or a methyl group.
The isochromene derivative is preferably a compound represented by the following formulas (1), (2) and (3), and the specific compound name is:
(1) 6-hydroxy-8- (2-hydroxy-4-methoxy-6-pentylphenoxy) -3-pentyl-1H-isochromen-1-one,
(2) 6,8-dihydroxy-3-pentyl-1H-isochromen-1-one, and (3) 8- (2,4-dihydroxy-6-pentylphenoxy) -6-hydroxy-3-pentyl-1H- Isochromen-1-one.

[2] Method for Producing Isochromene Derivatives The above phenol derivatives can be extracted and isolated mainly using oak moss, which is moss that grows on the epidermis of birch trees in Europe and America.
First, oak moss is extracted with a low polarity hydrocarbon solvent such as petroleum ether, benzene, hexane, etc., and the solvent is removed to obtain a solid extract (concrete) having a high wax content.
Next, in order to remove the wax and improve the solubility, the concrete is extracted again with an alcohol such as ethyl alcohol, and the alcohol is distilled off to obtain an extract (absolute).

Next, after a fraction having a high antibacterial activity is selected by a separation method based on the molecular size of the absolute, a target component that remarkably exhibits anti-legionella activity is isolated by a separation method based on the adsorptivity.
Examples of the separation method based on the molecular size include ultracentrifugation such as dialysis using a semipermeable membrane, ultrafiltration, gel filtration, and differential centrifugation. In the present invention, gel filtration chromatography using dextran gel obtained by reacting polysaccharide dextran with epichlorohydrin and cross-linking three-dimensionally to give a network structure, for example, “Sephadex Sephadex (trade name) manufactured by GE Healthcare Biosciences, Inc. ) "Is preferred.

  Adsorbents used for separation methods based on adsorptive properties such as activated carbon, silica gel, alumina, zeolite, synthetic adsorbents (styrene / divinylbenzene, (meth) acrylic, etc.), ion exchange resin, porous glass, cyclodextrin, etc. Can be mentioned. Among these, adsorption chromatography using a column packed with chemically bonded silica gel (octadecyl group bond or phenyl group bond) is preferable.

As a specific isolation method, first, a column packed with cross-linked polymer dextran gel using oak moss absolute using an organic solvent, preferably a mixed solvent of acetonitrile and methanol (acetonitrile: methanol is 10: 1 to 1:10). The first fraction is obtained by chromatography.
Next, as the second fraction, the first fraction was fractionated by column chromatography packed with chemically bonded silica gel using an acetonitrile aqueous solution (acetonitrile: water 10: 1 to 1:10). A method for obtaining the object can be exemplified. In addition, you may perform a 2nd fraction after going through the operation which fractionates a 1st fraction with the column chromatography filled with chemically bonded silica gel using methanol or methanol aqueous solution.

[3] Antibacterial agent The compound group used as the antibacterial agent in the present invention is an antibacterial agent against Legionella spp. Preferably composed of an isochromene derivative compound represented by the above formulas (1) to (3). Or it is used in combination of 2 or more types.

That is, the compound group used as an antibacterial agent in the present invention is preferably (1) 6-hydroxy-8- (2-hydroxy-4-methoxy-6-pentylphenoxy) -3-pentyl-1H-isochromene-1- ON, (2) 6,8-dihydroxy-3-pentyl-1H-isochromen-1-one, and (3) 8- (2,4-dihydroxy-6-pentylphenoxy) -6-hydroxy-3-pentyl- It is selected from 1H-isochromen-1-one, and these are used alone or in combination of two or more.
Some of these compounds are known to have antibacterial activity against various bacteria, but none of the antibacterial activity against Legionella is known.

Since the antibacterial agent of the present invention has strong antibacterial activity against Legionella, it can be used at a low concentration.
The use concentration of the antibacterial agent of the present invention can be selected according to the purpose of use, but is generally 0.1 to 100 ppm, preferably 0.5 to 50 ppm when used alone. Preferably it is used in the range of 1.0 to 10 ppm.
As a bactericide against Legionella spp., Chlorine is generally effective at about 1 ppm. When it is used repeatedly in a circulatory system facility, deactivation occurs due to the diffusion and decomposition of available chlorine. However, since the antibacterial agent of the present invention does not cause dispersal and decomposition, it can be said that the antibacterial agent is more effective than the same antibacterial activity as compared with chlorine.

  The isochromene derivative, which is an active ingredient of the antibacterial agent of the present invention, is effective when used as a composition in combination with other ingredients. As other components used in such a manner, various components are used depending on the purpose, but generally other fungicides, fragrances, dyes, surfactants and the like can be mentioned. Among these, a combination with a fragrance having antibacterial activity is particularly preferable.

  In the present invention, other antibacterial agents used in combination include surfactant fungicides such as quaternary ammonium salts, hydrazines and maleimides, isothiazoline compounds, phosphonium compounds, bromonitro compounds, pyrithione compounds and the like. Specifically, N, N-dimethyl-N, N-di-n-decylammonium chloride, N, N-dimethyl-N, N-di-n-octylammonium chloride, N, N-dimethyl-N-octyl -N-decylammonium chloride, N, N-dimethyl-N-isononyl-N-decylammonium chloride, N-hexadecyl-N, N, N-trimethylammonium chloride, N-hexadecyl-N, N, N-trimethylammonium bromide N, N-dimethyl-N-octadecyl-N- (3- (trimethoxytri ) Propyl) ammonium chloride,

Hydrazine, hydrated hydrazine, hydrazine hydrazine, maleimide, N-methylmaleimide, N-ethylmaleimide, citraconic acid imide, N-methylcitraconic acid imide, N-ethylcitraconic acid imide, 2,3-dimethylmaleic acid imide, N -Methyl-2,3-dimethylmaleimide, N-ethyl-2,3-dimethylmaleimide, N- (2-hydroxyethyl) maleimide, N-cyclohexylmaleimide, N-4-hydroxycyclohexylmaleimide, N- Phenylmaleimide, Np-hydroxyphenylmaleimide,

Np-hydroxybenzylmaleimide, 5-chloro-2-methyl-4-isothiazolin-3-one, 2-methyl-4-isothiazolin-3-one, 5-chloro-2-n-octylisothiazolin-3-one 1,2-benzisothiazolin-3-one, tetrakis (hydroxymethyl) phosphonium sulfate, tetrakis (hydroxymethyl) phosphonium phosphate, tetrakis (hydroxymethyl) phosphonium nitrate, tetrakis (hydroxymethyl) phosphonium hydrochloride, tetrakis ( Hydroxymethyl) phosphonium bromide, tetrakis (hydroxymethyl) phosphonium fluoride, 2-bromo-2-nitropropane-1,3-diol, 2,2-dibromo-2-nitropropane-1-ethanol, 2, 2-Dibromo-3-nitrop Pionamide, bis (2-pyridinethiol-1-oxide) copper salt, bis (2-pyridinethiol-1-oxide) zinc salt, bis (2-pyridinethiol-1-oxide) iron salt, 2-pyridylthio-1- Sodium oxide, pyrithione disulfide, glutaraldehyde, hydrogen peroxide, 1-bromo-3-chloro-5,5-dimethylhydantoin, methylenebisthiocyanate, sodium hypochlorite, 1,5-pentanedial,

3- (3,4-dichlorophenyl) -1,1-diphosphonic acid, 2,3,3-triiodoallyl alcohol, especially 2,2-dibromo-3-nitropropionamide, 2-bromo-2-nitropropane- 1,3-diol, 5-chloro-2-methyl-4-isothiazolin-3-one, and 2-methyl-4-isothiazolin-3-one are preferred. These bactericidal / antibacterial agents can be used alone or in combination with the isochromene derivative, and the blending amount thereof preferably does not exceed the blending amount of the isochromene derivative.

As another component used by this invention, it can manufacture, for example, using together the base, additive, etc. which are normally used by the bathing agent etc. which are shown below.
(1) Various fats and oils:
Avocado oil, almond oil, fennel oil, egoma oil, olive oil, orange oil, orange luffy oil, sesame oil, cacao butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acid, kukui nut oil, safflower oil, shea butter, liquid shea Fat, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, coconut oil, beef tallow, lard Examples include hydrogenated oils and fats (hardened oil and the like).

(2) Wax:
Examples include beeswax, mole, carnauba wax, whale wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, rice wax, squalene, squalane, pristane and the like.

(3) Mineral oil:
Examples include liquid paraffin, petrolatum, paraffin, ozokelide, ceresin, and microcrystalline wax.

(4) Fatty acids:
Natural fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, 12-hydroxystearic acid, undecylenic acid, tall oil fatty acid, lanolin fatty acid, isononanoic acid, caproic acid, 2-ethylbutanoic acid, isopentane Examples thereof include synthetic fatty acids such as acid, 2-methylpentanoic acid, and 2-ethylhexanoic acid.

(5) Alcohols:
Examples include ethanol, isopropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, natural alcohols such as cholesterol and phytosterol, and synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, and 2-octyldodecanol.

(6) Polyhydric alcohols:
Ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, 1,3-butylene glycol, 1 , 2-pentanediol, glycerin, diglycerin, pentaerythritol, sorbitol, mannitol and the like.

(7) Esters:
Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, diethyl phthalate, phthalate Examples include dibutyl acid, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate and the like.

(8) Metal soaps:
Examples include aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, and zinc undecylate.

(9) Gums, sugars or water-soluble polymer compounds:
Gum arabic, benzoin gum, danmar gum, guaiac gum, Irish moss, Karaya gum, tragacanth gum, carob gum, quinseed, agar, casein, lactose, fructose, sucrose or esters thereof, trehalose or derivatives thereof, dextrin, gelatin, pectin, starch, Carrageenan, carboxymethyl chitin or chitosan, hydroxyalkyl (C ₂ -C ₄ ) chitin or chitosan to which alkylene (C ₂ -C ₄ ) oxide such as ethylene oxide is added, low molecular chitin or chitosan, chitosan salt, sulfated chitin Or chitosan, phosphorylated chitin or chitosan, alginic acid or its salt, hyaluronic acid or its salt, chondroitin sulfate or its salt, heparin, ethylcellulose, methylcellulose, carboxymethylcellulose, carbo Polyalkylene oxide such as xyethyl cellulose, sodium carboxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, nitrocellulose, crystalline cellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, polyvinyl methacrylate, polyacrylate, polyethylene oxide and polypropylene oxide or Examples thereof include a crosslinked polymer, a carboxyvinyl polymer, and polyethyleneimine.

(10) Surfactant:
Anionic surfactant (alkyl carboxylate, alkyl sulfonate, alkyl sulfate ester salt, alkyl phosphate ester salt), cationic surfactant (alkylamine salt, alkyl quaternary ammonium salt), amphoteric surfactant [carboxylic acid Type amphoteric surfactant (amino type, betaine type), sulfate type amphoteric surfactant, sulfonic acid type amphoteric surfactant, phosphate ester type amphoteric surfactant], nonionic surfactant (ether type nonionic surfactant) Surfactant, ether ester type nonionic surfactant, ester type nonionic surfactant, block polymer type nonionic surfactant, nitrogen-containing type nonionic surfactant), other surfactants (natural surfactants, Derivatives of protein hydrolysates, polymer surfactants, surfactants containing titanium and silicon, fluorocarbon surfactants ), And the like.

(11) Various vitamins:
Vitamin A group: retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, lycopene (provitamin A),
Vitamin B group: thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acids, pantothenic acids, biotins, choline, inositols ,
Vitamin C group: Vitamin C acid or its derivative,
Vitamin D group: Ergocalciferol (vitamin D2), Cholecalciferol (vitamin D3), Dihydrotaxosterol,
Vitamin E group: Vitamin E or its derivatives, ubiquinones,
Vitamin K group: phytonadione (vitamin K1), menaquinone (vitamin K2), menadione (vitamin K3), menadiol (vitamin K4),
In addition, essential fatty acids (vitamin F), carnitine, ferulic acid, γ-oryzanol, orotic acid, vitamin Ps (rutin, eriocitrin, hesperidin), vitamin U and the like are exemplified.

(12) Various amino acids:
Valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, arginine, ornithine, histidine Examples thereof include amino acid derivatives such as sulfates, phosphates, nitrates, citrates, and pyrrolidone carboxylic acids.

(13) Various additives derived from plant or animal materials:
These are processed in a conventional manner according to the product type and form to be added (for example, pulverization, milling, washing, hydrolysis, fermentation, purification, pressing, extraction, fractionation, filtration, drying, pulverization) , Granulation, dissolution, sterilization, pH adjustment, deodorization, decolorization and the like are arbitrarily selected and combined), and may be arbitrarily selected from various materials.

  The solvent used for the extraction of the plant or animal material may be selected in consideration of the intended purpose and type of the product to be provided, or the processing to be performed later. It is desirable to use one or a mixture of two or more selected from organic organic solvents (for example, ethanol, propylene glycol, 1,3-butylene glycol, etc.). However, when it is not preferable to contain an organic solvent depending on the application, it is sufficient to use only water or ethanol that can be easily removed after extraction, and it may be used alone or in any mixture with water. The thing extracted and extracted may be sufficient.

  In addition, when the additive derived from the plant or animal material is used for systemic or topical external preparations, cosmetics, including the protection of skin and hair, moisturizing, improvement of touch / feel, imparting flexibility, Relaxation of stimulation, stress reduction by fragrance, cell activation (prevention of cell aging), suppression of inflammation, improvement of skin / hair quality, prevention of rough skin and its improvement, hair growth, hair growth, prevention of hair loss, imparting luster, cleaning effect In addition to cosmetic effects such as fatigue relief, blood flow promotion, and warm bath effects, effects such as flavoring, deodorizing, thickening, preserving, and buffering can also be expected.

(14) Fragrance:
Natural animal fragrances such as musk, civet, castorium, ambergris, anise essential oil, angelica essential oil, ylang ylang essential oil, iris essential oil, fennel essential oil, orange essential oil, cananga essential oil, caraway essential oil, cardamom essential oil, guayakwood essential oil, cumin essential oil, black letter Essential oil, cinnamon essential oil, cinnamon essential oil, geranium essential oil, copaiba balsam essential oil, coriander essential oil, perilla essential oil, cedarwood essential oil, citronella essential oil, jasmine essential oil, gingergrass essential oil, cedar essential oil, spearmint essential oil, western peppermint essential oil, dairy perfume essential oil, tuberose Essential oil, dichroic essential oil, orange flower essential oil, winter green essential oil, trubalsum essential oil, patchouli essential oil, rose essential oil, palmarosa essential oil, cocoon essential oil, hiba essential oil, sandalwood essential oil, petit gren essential oil, bay essential oil, vetiver essential oil, bergamot essential oil, Examples include plant flavors such as rubalsum essential oil, boad rose essential oil, melamine essential oil, mandarin essential oil, eucalyptus essential oil, lime essential oil, lavender essential oil, linaloe essential oil, lemongrass essential oil, lemon essential oil, rosemary essential oil, Japanese mint essential oil .

  Synthetic perfume ingredients include aliphatic hydrocarbons, terpene hydrocarbons, aromatic hydrocarbons and other hydrocarbons, aliphatic alcohols, terpene alcohols, aromatic alcohols and other alcohols, aliphatic ethers, aromatic ethers and other ethers Oxides, aliphatic oxides, terpene oxides, aliphatic aldehydes, terpene aldehydes, hydrogenated aromatic aldehydes, aldehydes such as thioaldehydes, aromatic aldehydes, aliphatic ketones, terpene ketones, hydrogenated aromatics Ketones such as aromatic ketones, aliphatic cyclic ketones, non-benzene aromatic ketones, aromatic ketones, acetals, ketals, phenols, phenol ethers, fatty acids, terpene carboxylic acids, hydrogenated aromatic carboxylic acids, Acids such as aromatic carboxylic acids, acid amides, aliphatic lactones Lactates such as macrocyclic lactones, terpene lactones, hydrogenated aromatic lactones, aromatic lactones, esters such as aliphatic esters, furan carboxylic acid esters, aliphatic cyclic carboxylic acid esters, aromatic carboxylic acid esters And nitrogen-containing compounds such as nitromusks, nitriles, amines, pyridines, quinolines, pyrrole and indole.

  More specific examples of synthetic fragrances include “synthetic fragrance chemistry and product knowledge” (1996, published by Kagaku Kogyo Nippo Inc., Motoichi Into) and “Perfume and Flavor Chemicals” (1969, The fragrance | flavor etc. which were described in the STEPHANARCTAMDER etc. can use it conveniently.

The main fragrances are shown below. Aldehydes C _{6 to} C ₁₂ , anisaldehyde, acetal R, acetophenone, acetyl cedrene, adxal, allyl amyl glycolate, allyl cyclohexane propionate, alpha damascon, beta damascon, delta damascon, ambret lid, ambroxan , Amylcinnamic aldehyde, amylcinnamic aldehyde dimethyl acetal, amyl valericate, amyl salicylate, isoamyl acetate, isoamyl salicylate, auranthiol, acetyl eugenol, bacdanol, benzyl acetate, benzyl alcohol, benzyl salicylate, bergamyl acetate, vol Nyl acetate, butyl butyrate, para tertiary butyl cyclohexanol, para tertiary butyl Rohexyl acetate, ortho-tertiary butyl cyclohexanol, benzaldehyde,

Benzyl formate, Kali olefin, Kashmelan, Carvone, Cedro amber, Cedryl acetate, Cedrol, Cestrid, Cinamic alcohol, Cinamic aldehyde, Cisjasmon, Citral, Citral dimethyl acetal, Citrusal, Citronellol, Citronellyl acetate, Citronellyl Formate, citronellyl nitrile, cyclaset, cyclamenaldehyde, cyclaprop, caron, coumarin, cinnamyl acetate, delta C ₆ -C ₁₃ lactone, dimethylbenzyl carbinol, dihydrojasmon, dihydrolinalol, dihydromyrsenol, dimethol,

Jimirusetoru, diphenyl oxide, Echiruwanirin, Yugenoru, Furuiteto, fenchyl alcohol, phenylethyl phenylacetate, Garakisoriddo, gamma C ₆ -C ₁₃ lactones, geraniol, geranyl acetate, geranyl formate, geranyl nitrile, Hedion, ocean propanal, heliotropine, cis -3-hexenol, cis-3-hexenyl acetate, cis-3-hexenyl rusalicylate, hexylcinnamic aldehyde, hexyl salicylate, hyacinth dimethyl acetal, hydrotropic alcohol, hydroxycitronellal, indole, isobornyl acetate, isocyclo Citral, iso-E super, isoyuugenol, isononyl acetate, isobutyl quinoline, jasma Le, jasmonate lactone, jazz Mofi orchid, Koabon, Rigusutoraru, lilial, lime oxide, limonene, linalool, linalool oxide, linalyl acetate, and the like.

The use of the antibacterial agent of the present invention is particularly effective as an industrial disinfectant such as a cold / hot water system such as a circulating hot water of a 24-hour bath, a heat storage water system, a cooling tower, a circulating cooling water of a refrigeration apparatus.
Since the antibacterial agent of the present invention is not inactivated or decomposed, it can control Legionella spp. Surviving in water circulated for a long time and prevent its growth.
Therefore, it can be widely applied to household or commercial bathing agents (bathing agents) and bathtub cleaning agents.

The present invention will be described in more detail with reference to the following examples. However, the following examples are used only for illustrative purposes, and the present invention is not limited thereto.
[Production Example 1] Preparation of sample (1) Raw material (oak moss extract):
Oak moss absolute manufactured by Charabot, France was used. The absolute is extracted by extracting oak moss (Evernia prunastri (L.) Arch.) (Tsunomatagoke) that grows on oak trees with low-polarity hydrocarbon solvents such as petroleum ether, benzene, hexane, etc. It is obtained by extracting with ethyl alcohol and removing the alcohol.

(2) First fraction “Sephadex LH-20 (trade name)” manufactured by GE Healthcare Biosciences [matrix: hydroxypropylated Sephadex G-25, particle size: 18 to 111 μm (when powdered), 27 to 163 μm (when methanol swells, fractionation range: 100 to 4000 Da) on an open column packed with 30 × 500 mm, 2.24 g of raw material Oak moss absolute is placed, and acetonitrile: methanol = 10: 1 solvent at a rate of 2 ml per minute And fractionated into fractions 1 to 9.
The amount of the extract of each fraction after solvent removal is as follows.

  Among these, anti-Legionella activity was found in fractions 2-5 and 8 and used for the next fractionation.

(3) Secondary fraction (a) Isolation of compound (P) About 12 mg of fraction 3 (total 315.6 mg) was subjected to liquid chromatography (silica gel chromatography with phenyl groups attached to VARIAN "Bond Elut PH (product Name))), and 10 ml each of 20% methanol, 40% methanol, 60% methanol, 80% methanol, and 100% methanol was allowed to flow.
This was repeated and fractionated. The amount of each spilled fraction is as follows.

Among these, strong anti-Legionella activity was observed in the 60% methanol fraction to 80% methanol fraction, and the peak of the compound (P) was observed with a UV 280 nm detector.
The liquid fraction (C ₁₈ (octadecyl) group was 142.4 mg, which was obtained by combining the 60% methanol fraction (62.6 mg) and the 80% methanol fraction (79.8 mg) of the above fraction 3 obtained in the second fraction. Introduced into Shiseido “CAPCELL PAK MG II (trade name)”, 10 × 250 mm column), which is a bound silica gel chromatograph, flowing 4 ml of solvent at a ratio of acetonitrile: water = 65: 35 per minute, and using a UV 280 nm detector While monitoring, a flow of 30 minutes after the start was collected to obtain Compound (P).

(B) Isolation of Compound (K2) Fraction 5 (total amount 259.8 mg) obtained in the first fractionation was subjected to liquid chromatography (silica gel chromatography combined with C ₁₈ (octadecyl) group “CAPCELL PAK MG manufactured by Shiseido” II (trade name) ”, 10 × 250 mm column) and acetonitrile / water = 38: 62 at a rate of 35 minutes and 65:35 at a rate of 15 minutes at a rate of 4 ml / min. While monitoring, a fraction of 46.5 minutes after the start was collected to obtain compound (K2).

(C) Isolation of Compound (L) “CAPCELL PAK” manufactured by Shiseido Co., Ltd., which is a silica gel chromatograph with fraction (86.5 mg in total) of fraction 8 obtained in the first fractionation combined with a C ₁₈ (octadecyl) group MG II (trade name), 4.6 × 250 mm column), flowing 1 ml per minute under the conditions of acetonitrile: water = 50: 50 for 15 minutes, 80:20 for 15 minutes, and using a UV 280 nm detector. While monitoring, a stream 24 minutes after the start was collected to obtain compound (L).

The total amount of each compound obtained and its properties are as follows.

The structure of each compound obtained was identified by instrumental analysis.
[1] Compound (P): 6-hydroxy-8- (2-hydroxy-4-methoxy-6-pentylphenoxy) -3-pentyl-1H-isochromen-1-one (1) ¹ H NMR and ¹³ C NMR Spectral analysis

(2) Mass spectrometry
MS m / z 440 (M ⁺ )

[2] Compound (K2): 6,8-dihydroxy-3-pentyl-1H-isochromen-1-one (1) ¹ H NMR and ¹³ C NMR spectrum analysis

(2) Mass spectrometry
MS m / z 248 (M ⁺ )

[3] Compound (L): 8- (2,4-dihydroxy-6-pentylphenoxy) -6-hydroxy-3-pentyl-1H-isochromen-1-one (1) According to ¹ H NMR and ¹³ C NMR spectra analysis

(2) Mass spectrometry
MS m / z 426 (M ⁺ )

[Test Example 1]
(1) Strains used and culture:
The strain was Legionella peneumophila JCM7571 (Philadelphia 1) strain, and cultured in a buffered charcoal yeast extract agar supplemented with 0.1% α-ketoglutarate (BCYE-α, Becton Dickinson, MD) plate agar medium at 37 ° C. for 72 hours.

(2) Antibacterial test medium:
A buffered yeast extract supplemented with 0.1% α-ketoglutarate (BCYE-α) liquid medium was used as an antibacterial test medium.

(3) Preparation of Test Bacterial Suspension For the measurement of minimum inhibitory concentration (MIC), a test bacterial suspension prepared as follows was used as a stock solution and appropriately diluted. L. peneumophia cells cultured on BCYE-α plate agar medium were added 4 mL of sterilized saline directly on the medium, made into a suspension while stirring with a cornagome pipette, and then transferred to a sterile test tube. The mixture was stirred with a vortex mixer to obtain a uniform suspension. Furthermore, it diluted with the sterilized physiological salt so that the light absorbency in 600 nm might be set to 0.1, and was set as the test microbe suspension stock solution.
This suspension showed 1.7 × 10 ⁸ CFU / mL as a result of colony forming unit (CFU) measurement.

(4) Antibacterial test: Measurement of MIC The MIC measurement was basically based on a modified method based on the micro liquid dilution method of the US Clinical Laboratory Standards Institute (CLSI) standard method, but L. pneumophila Was sensitive to poly (oxyethylene) sorbitan monooleate (trade name “Tween 80”) used as a dispersant for the sample, so that the sample was diluted without using a dispersant.
A sample diluted 4-fold with dimethyl sulfoxide (DMSO) was diluted and stirred with BCYE-α to form a solution or suspension to prepare BCYE-α containing 1.0 (%, V / V) sample. The obtained 1.0 (%, V / V) sample-containing BCYE-α is 1.0 to 0.0020 (%, V) using a deep well plate (“BM6022” manufactured by BM Instruments Co., Ltd.) and BCYE-α. / V). The test bacterial suspension was prepared by diluting the test bacterial suspension stock solution with BCYE-α so that the concentration became 6.0 × 10 ⁵ CFU / mL.

50 μL of diluted test solution of each sample and 50 μL of test bacteria suspension are dispensed into each well of a 96-well microplate (“BM6000” manufactured by BM Instruments Co., Ltd.), stirred and mixed, and then a plate seal (Sanko Junyaku Co., Ltd.) Each well was sealed using “SJ103-40” manufactured by the company and cultured at 37 ° C. for 48 hours. Based on the CLSI standard method, the MIC of each sample was determined by visually observing the presence or absence of cells that grew after culture and settled or floated. Moreover, the test repeated the same test twice.
For comparison, the same test was performed on oak moss absolute, cinnamic aldehyde and chlorhexidine gluconate, which are known to have anti-legionella activity.

  Table 1 shows the MIC measurement results for L. pneumophila of each compound and comparative substance.

  From the results in Table 1, the usefulness of the antibacterial agent of the present invention was shown.

  Next, the application range of the present invention will be described in more detail with reference to application examples, but the present invention is not limited to these application examples. The antimicrobial agent-containing product of the present invention was produced according to each formulation. All formulation examples are shown in parts by weight.

[Application Example 1] Antibacterial spray agent In accordance with Formulation Example 1, 0.2% of silicic acid anhydride is blended in a spray base prepared by mixing and dissolving a perfume and the compound P of the present invention at room temperature in ethyl alcohol, and in a spray container. The antibacterial spray agent of the present invention was prepared by encapsulating LPG and isopentane so that the pressure of the solution was 8 kg / cm ² or less at 35 ° C.

[Application 2] Bath agent a
In accordance with Formulation Example 2, a bath additive base prepared by adding perfume and the like to sodium sulfate and sodium hydrogen carbonate was prepared, and 0.2% of Compound K2 of the present invention was blended. Manufactured.

[Application Example 3] Bath agent b
In accordance with Formulation Example 3, perfume in ethyl alcohol, compounded into a bath base prepared by mixing and dissolving the compound L of the present invention at room temperature, 0.1% polyoxyethylene hydrogenated castor oil, 30.0% purified water, It was set as the bath agent b of this invention.

[Application Example 4] Disinfectant According to Formulation Example 4, a nonionic surfactant was added to ethyl alcohol and dissolved. To this, the compound L of the present invention dissolved in ethyl alcohol was added and mixed and dissolved. Further, purified water was added and dissolved to obtain the cooling tower disinfectant of the present invention.

[Application Example 5] Bath Cleaner According to Formulation Example 5, a cationic surfactant was added to fatty acid amidopropyl betaine and dissolved. To this, each equivalent mixture of chelating agent and compounds P and L of the present invention was added and mixed and dissolved. Furthermore, purified water was added and dissolved to obtain the bathtub cleaning agent of the present invention.

The antibacterial agent of the present invention can provide an anti-Legionella bacterium that has no side effects even when used on a daily basis, is safe, and can strongly inhibit the growth of Legionella bacterium.
In addition, by adding the Legionella antibacterial composition containing the antibacterial agent of the present invention, it survives in a cooling tower, refrigeration equipment circulating cooling water, chilled hot water systems such as 24-hour bath circulating hot water, and heat storage water systems. It is possible to control Legionella spp. And prevent its growth.

Claims (7)

The following formula (I):

[Wherein R ¹ is a hydrogen atom or the following formula (II):

(Wherein R ² represents a hydrogen atom or a linear or branched alkyl group having 1 to 6 carbon atoms)]
The antibacterial agent with respect to Legionella genus bacteria characterized by including the isochromene derivative represented by these as an active ingredient. The isochromene derivative is represented by the following formula (1):

The antibacterial agent according to claim 1, which is a compound of The isochromene derivative is represented by the following formula (2):

The antibacterial agent according to claim 1, which is a compound of The isochromene derivative is represented by the following formula (3):

The antibacterial agent according to claim 1, which is a compound of   A bath preparation comprising the antibacterial agent according to any one of claims 1 to 4.   An industrial disinfectant comprising the antibacterial agent according to any one of claims 1 to 4.   A bath cleaner comprising the antibacterial agent according to any one of claims 1 to 4.