JP2001131061A - Bactericide for vancomycin resistant enterococcus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a bactericide for a vancomycin resistant enterococcus. SOLUTION: This bactericide contains hinokithiol, its metal complex or their salts as an active ingredient. Further, it is preferable to contain at least 1 kind of extract of a plant selected from Aloe, a green tea, low striped bamboo and Houttuynia cordata. An aqueous solution type containing water as a carrier is preferably used.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to the disinfection of vancomycin-resistant enterococci (VRE) that are resistant to vancomycin hydrochloride, the disinfection of medical sites to which vancomycin-resistant enterococci are adhered, and the vancomycin-resistant enterococci inhabit the intestine. The present invention relates to a fungicide for vancomycin-resistant enterococcus, which is used for intestinal sterilization of animals (humans, cattle, chickens, etc.) which are considered to be affected.

[0002]

BACKGROUND OF THE INVENTION Vancomycin hydrochloride acts bactericidal on most bacteria and bacteriostatic on enterococci. In particular, it has been approved as an antibiotic against MRSA in Japan because it is effective against mesicillin-resistant staphylococci (MRSA), which has recently become a problem as a hospital-acquired bacterium.

[0003] Since vancomycin hydrochloride has been known and used as a therapeutic agent for MRSA, bacteria resistant to vancomycin hydrochloride have appeared. For this reason,
At present, a substance having a bactericidal and antibacterial activity against vancomycin-resistant enterococci has been desired.

[0004] The present invention has been made in view of such circumstances, and an object of the present invention is to provide a fungicide for vancomycin-resistant enterococci.

[0005]

DISCLOSURE OF THE INVENTION The present invention has completed the present invention by finding that hinokitiol, which is known as a bactericidal component having a relatively wide antibacterial spectrum, is also effective against vancomycin-resistant bacteria.

That is, the fungicide for vancomycin-resistant enterococci of the present invention contains hinokitiol, a metal complex thereof, or a salt thereof as an active ingredient.

Further, it preferably contains at least one extract selected from the group consisting of aloe, green tea, kuma bamboo grass, and dokudami, and further contains persimmon leaves, pine, cedar, sweet tea, perilla, wasabi and madder , Ume, garlic, peppermint, mugwort, salmon, rhubarb, thistle, mint, loquat, purple, lavender, lemongrass, and mint.

The vancomycin-resistant enterococci bactericide of the present invention is preferably an aqueous solution containing water as a carrier.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The fungicide for vancomycin-resistant enterococci of the present invention contains hinokitiol, a metal complex thereof, or a salt thereof as an active ingredient.

[0010] Hinokitiol is also referred to as β-tsuyaprisin, and is contained in essential oils such as Taiwan Hinoki, one of Aomori-grown Hiba (Asnaro), and one of the smells. For the composition according to the present invention, hinokitiol derived from a natural product may be used, or a chemically synthesized product may be used. Specifically, a commercially available product sold by Takasago International Corporation or Osaka Organic Chemical Industry can be used.

Examples of the metal complex of hinokitiol include metal complexes of hinokitiol with zinc, copper, iron, calcium, magnesium, barium, tin, cobalt, titanium, vanadium and the like. The ratio of hinokitiol to metal is not particularly limited, but usually a hinokitiol: metal molar ratio of 2: 1 or 3: 1 is preferably used.

The salts of hinokitiol or hinokitiol metal complexes include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; transition metal salts such as copper salt and zinc salt; diethanolamine Salt, 2-amino-2-ethyl-1,3-
Alkanolamine salts such as propanediol salt and triethanolamine salt; organic salts such as morpholine salt, piperazine salt and heterocyclic amine salt such as piperidine salt, ammonium salt, basic amine salt such as arginine salt, lysine salt and histidine salt. And the like.

These hinokitiols or metal complexes thereof, or salts thereof, may be used alone or in combination of two or more. These may be appropriately selected and used in accordance with the form of the composition, but when used as a form that can be directly taken into the human body, it is preferably used as hinokitiol. On the other hand, a metal complex of hinokitiol or a salt of a metal complex is superior in light resistance to hinokitiol, and therefore, when the composition of the present invention is used for an application requiring weather resistance, a metal complex or metal complex of hinokitiol is used. It is preferable to use a salt of

[0014] These hinokitiols or metal complexes thereof, or salts thereof (hereinafter referred to as "hinokitiol or the like" when they are not distinguished from each other) exhibit excellent antibacterial activity against vancomycin-resistant enterococci. Hinokitiol is known to have a wide range of antibacterial effects, including aerobic bacteria such as tuberculosis and Salmonella typhoid, anaerobic bacteria such as clostridia, salmonella bacteria causing food poisoning, and Escherichia coli. It has been found by the present inventors to be bactericidal against enterococci.

The bactericide for vancomycin-resistant enterococci according to the present invention contains hinokitiol, which exhibits bactericidal activity effective against vancomycin-resistant enterococci, as an essential active ingredient, and 50 to 100 μg of hinokitiol alone is used.
Effective bactericidal properties can be exhibited at a certain level.

The fungicide for vancomycin-resistant enterococci according to the present invention may further comprise at least one extract selected from the group consisting of aloe, green tea, Kuma-basa and dokudami (hereinafter referred to as "aloe, etc." Extract "). This is because these extracted components have the function of increasing the antibacterial activity of hinokitiol against vancomycin-resistant enterococci, and can reduce the amount of hinokitiol required to inhibit the growth of vancomycin resistance. Hinokitiol generally has a solubility in water of:
Although it is said that the limit is 0.2% by mass, an aqueous solution of about 1% by mass can be easily obtained by using together with an extract such as aloe. Therefore, when a bactericide is used as an aqueous solution, it is particularly preferable to coexist an extract such as aloe.

The aloe extract is an extract obtained by extracting a jelly-like body (leaf) of aloe leaves mainly by a squeeze extraction method and applying heat to concentrate and stabilize the extract. Instead of such an aloe extract, an anthraquinone derivative, which is a main component, such as aloin or barberloin, may be used. The aloe extract includes aloe-emodin, aloecin, aloeenin and the like in addition to aloin and barberloin.

As the green tea extract, a liquid obtained by extracting ground green tea with hot water, purifying and concentrating the green tea is used. The main component of green tea extract is tea polyphenol. Tea polyphenol is a general term for compounds having a plurality of phenolic hydroxyl groups in the molecule, and has catechin, epicatechin, gallocatechin, epigallocatechin, epigatechin gallate, epigallocatechin gallate, and the like as main components.

The extract of Kumasa is extracted by a low temperature and high pressure extraction method.
Obtained by extracting Kumasa. The low-temperature and high-pressure squeezing extraction method is a method of extracting Kumasa bamboo without increasing the temperature using a mechanical device set at a high pressure, and a liquid obtained by concentrating the liquid squeezed out at that time becomes Kumasakasa extract. Kumasasa is a kind of Sasa of the Poaceae widely distributed in Japan and China. The Kumasasa extract contains lignin residues, reducing sugars, and sugars such as glucose, in addition to triterpenol (β-amyrin / Frieden), which is the main component. Instead of Kumasasa extract,
Mixtures of these synthetic products can also be used.

Dokudami is a perennial herb that is distributed in Japan, Thailand, China, Himalayas and Java, and is found in mountains and gardens. The extract of Dokudami is extracted by a method called low-temperature and high-pressure squeezing extraction method, similar to Kumasasa. Dokudami extract includes quercitrin, afzenin (afze
nin), hyperin, rutin, chlorogenic acid, β-sitosterol, cis and trans-N- (4-hydroxystyryl). A mixture of these synthetic products can be used in place of the extract of Kumasasa.

The extract of aloe, green tea, Kumasa and Dokudami may be used alone, but preferably used in combination of two or more, and more preferably contains all of these extracts.

The content ratio of the extract such as hinokitiol and aloe is 40 μg to 300 μg for aloe and 40 μg to 300 μg for green tea per 100 μg of hinokitiol.
g, Kumasa, 20μg ~ 50μg, Dokudami 20μg ~
A range of 50 μg is preferred. More preferably, aloe,
This is a case in which all of the extracts of green tea, Kumasa and Dokudami are included, and the total content of extracts of aloe, green tea, Kumasa and Dokudami is 3 to 3.5 (mass ratio) with respect to Hinokitiol 1.

The fungicides for vancomycin-resistant enterococci according to the present invention include persimmon leaves, pine, cedar, amacha zuru, perilla,
The extract may contain wasabi, madder, prunus, garlic, peppermint, mugwort, salmon, rhubarb, thistle, peppermint, loquat, purple lavender, lavender, lemongrass, and forsythia, and propolis extracted from honey. These can alleviate the unique smell and bitterness of hinokitiol and increase the solubility of hinokitiol in water without impairing the antibacterial activity of hinokitiol against vancomycin-resistant enterococci.

The fungicide of the present invention contains an extract such as hinokitiol, preferably aloe as an active ingredient, and further contains a plant extract as required, and a carrier suitable for the form of the fungicide.

As the carrier, any carrier can be used as long as it does not cause a chemical change of the active ingredient, and it may be a liquid or a solid. In addition, the carrier is not limited to one type, and two or more types can be used as a mixture as needed.

The liquid carrier may be any solvent that can serve as a solvent and dispersion medium for hinokitiol, such as water; alcohols such as methanol and ethanol; ketones such as acetone and methyl ethyl ketone; fatty acid esters; benzene, toluene and xylene. Aromatic amides; acid amides such as dimethylformamide; and higher fatty acids.

Of these, water is preferably used for applications in which a large amount of a disinfectant is used, such as spraying or coating, or for drinking. Hinokitiol is generally known as poorly soluble in water, and hinokitiol alone can only dissolve about 0.2% by mass. However, by using an extract such as aloe in combination, an aqueous bactericide of about 1% by mass can be obtained. Can be. When water is used as the carrier, a surfactant such as glycerin fatty acid ester and a plant-based emulsifier such as Kirayasaponin may be used in combination, if necessary. By containing these, an aqueous solution of a bactericide having a hinokitiol concentration increased to 10% by mass can be obtained.

When water is used as a carrier, the preferred composition of the aqueous bactericide is 50 to 100 g of hinokitiol and 20 to 100 g of aloe per 1000 g of water.
g, green tea 20 μg to 100 g and Kumasa 10 μg to 50 g are appropriately selected depending on the content of hinokitiol in water, the presence or absence of a surfactant, and the like.

For disinfection, alcohols can be preferably used as the liquid carrier. For alcohol,
Even with hinokitiol alone, a bactericide having a hinokitiol content of up to about 5% by mass can be obtained. Increasing the concentration of hinokitiol in the liquid disinfectant means that the required dosage and amount used can be reduced,
The use of a large amount of disinfectant, such as for application and disinfection, is also preferred because it can be diluted when used.

Examples of the solid carrier include powder carriers, tablet carriers, and substances conventionally known as ointment bases, such as oils and fats, saccharides, proteins, esters, inorganic salts, and minerals.

The bactericide of the present invention can be used as a medicine for patients infected with vancomycin-resistant enterococci, can be used for sterilization and disinfection at medical sites, daily necessities food manufacturing sites, etc. And used as an intestinal fungicide for livestock and humans.

When the bactericide of the present invention is used as a drug (anti-vancomycin agent), a carrier is appropriately selected, and a tablet,
It can take various forms such as pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, injections (solutions, suspensions, etc.), eye drops, ointments and the like.

For example, in the case of tablets, excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, and silicic acid;
Binders such as water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinyl virolidone; dry starch,
Disintegrants such as sodium alginate, agar powder, laminaran powder, sodium hydrogen carbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose; sucrose, stearin, cocoa butter, hydrogenated Oils and other disintegration inhibitors; quaternary ammonium bases, absorption promoters such as sodium lauryl sulfate; humectants such as glycerin and starch; starch, lactose, kaolin, bentonite,
Tablets containing adsorbents such as colloidal silicic acid; lubricants such as purified talc, stearates, powdered boric acid, and polyethylene glycol can be prepared. Tablets are tablets coated with ordinary skin if necessary, such as sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets or double tablets,
Multiple tablets can be made.

In the case of pills, glucose, lactose, starch,
Excipients such as cocoa butter, hardened vegetable oil, kaolin, talc;
Binders such as gum arabic powder, tragacanth powder, gelatin, and ethanol; and disintegrants such as laminaran and agar can be contained.

In the case of suppositories, polyethylene glycol, cocoa butter, higher alcohol, esters of higher alcohol, gelatin, semi-synthetic glyceride and the like are used.

In the case of capsules, the active ingredient may be mixed with the various carriers exemplified above and then filled in a hard gelatin capsule or a soft gelatin capsule according to a conventional method.

When used as an injection, the carrier can be any of those used in the art as a diluent for the active ingredient. For example, water, ethyl alcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol,
Polyoxyethylene sorbitan fatty acid esters and the like can be used. Further, a sufficient amount of salt, glucose or glycerin for preparing an isotonic solution may be contained, and ordinary solubilizers, buffers, soothing agents and the like may be added.

When used as an ointment, oily bases known in the art can be widely used. Specifically, peanut oil, sesame oil, soybean oil, safflower oil,
Avocado oil, sunflower oil, corn oil, rapeseed oil,
Menzitsu oil, castor oil, camellia oil, coconut oil, olive oil, poppy oil, cocoa oil, beef tallow, lard, wool oil, etc., mineral oil such as petrolatum, paraffin, silicone oil, squalane, isopropyl myristate, n -Butyl myristate, isopropyl linoleate, acetyl ricinolate, stearyl ricinolate, diethyl sebacate,
Higher fatty acid esters such as diisopropyl adipate, cetyl alcohol, stearyl alcohol, salami beeswax, spermaceti, spermaceti, higher fatty alcohols and waxes, higher fatty acids such as stearic acid, oleic acid, and palmitic acid; Examples thereof include mono-, di-, and triglyceride mixtures of saturated or unsaturated fatty acids.

The fungicide of the present invention includes an active ingredient such as hinokitiol and aloe extract, an extract such as persimmon leaf added as needed, a carrier selected depending on the form,
Furthermore, if necessary, conventionally used additives such as metal soaps, animal extracts, vitamins, hormonal agents, medicinal agents such as amino acids, pigments, dyes, pigments, fragrances, ultraviolet absorbers, humectants, humectants, A tackifier, an antioxidant, a sequestering agent, a pH adjuster and the like can be appropriately compounded.

When used for sterilization or disinfection at medical sites or food manufacturing sites, etc., spray types for directly spraying on medical instruments, floors, walls, ceilings, etc .; application types for application; cloths, paper towels, wet tissues, It is preferable to use an impregnating-type disinfectant that impregnates a cloth for wiping into a towel or the like. When the object of disinfection and sterilization is clothing, food, tableware, gauze, sheets, diapers, curtains, and the like, it is preferable to use it by adding it to a cleaning agent, a finishing agent, and the like.

When used as a spray type, a coating type, or an impregnating type, the carrier may be a liquid carrier (water, alcohols,
Esters and higher fatty acids). When used by adding to a cleaning agent, a finishing agent, or the like, a liquid carrier that is compatible with the cleaning solution may be used, or a solid carrier that can be dissolved in the cleaning solution may be used.

When the fungicide of the present invention is used for spraying, coating, impregnating, or the like, the fungicide may be further diluted with a solvent or water as appropriate.

The fungicide for vancomycin-resistant enterococci of the present invention can be used not only for killing vancomycin-resistant enterococci present in the human intestine but also for animals other than humans, such as cattle,
It is also effective for disinfecting vancomycin-resistant enterococci present in the intestines of domestic animals such as pigs, birds, sheep and the like, chickens, turkeys, quails and other pets, dogs, cats and other pets. In this case, a carrier that does not cause harm when eaten, specifically, a carrier used in pharmaceuticals and the like is used, and water is preferably used.

When used as a bactericide in the intestine of animals, it is easy to add it to animal feed and feed it.

[0045]

EXAMPLES [Preparation of compositions for anti-vancomycin-resistant cocci] Two types of compositions having the compositions shown in Table 1 were prepared.

[0046]

[Table 1]

[Enterococci used] Six types of bacteria shown below were used. Enterococcus Faecalis KI
H-C233 (from patient's vaginal mucosa) MIC (μg / ml): VCM2, ABPC1, PCG
4, CEZ> 32, CMZ> 32, EM> 32, GM>
32, IPM1 Enterococcus faecium KIH
-234 (from patient's vaginal mucosa) MIC (μg / ml): VCM0.5, ABPC16,
PCG16, CEZ> 32, CMZ> 32, EM> 3
2, GM> 32, IPM1 Enterococcus faecium KIH
MIC (μg / ml): VCM500, ABCC4, P
CG16, CEZ> 32, CMZ> 32, EM> 32,
GM> 32, IPM8 Enterococcus gallinarum
KIH-241 (derived from domestic chickens) MIC (μg / ml): VCM8, ABCP4, PCG
1, CEZ> 32, CMZ> 32, EM> 1, GM> 3
2, IPM1 Enterococcus faecalis standard bacterium IFO-12965 Enterococcus faecium standard bacterium IFO-3535

VCM is vancomycin, A
BPC is aminobenzylpenicillin, PCG is benzylpenicillin, CEZ is cefazolin, C
MZ is Cefmetazole, EM is Erythromycin, GM is Zentamicin (Gen
tamycin), IPM stands for Imipenem. Vancomycin has a maximum blood concentration of 6 μm
/ Ml, the minimum inhibitory concentration is 5μ
If m / ml or more, vancomycin resistance can be said. Therefore, among the above-mentioned bacteria, are vancomycin-sensitive bacteria, and-are vancomycin-resistant bacteria.

[Minimum inhibitory concentration of hinokitiol against vancomycin-resistant enterococci] The minimum inhibitory concentration (MIC) was determined by an agar plate dilution method.

In the liquid for Mueller Hinton agar medium,
A diluent of the composition prepared above was added and solidified at room temperature to prepare an agar medium. Hinokitiol concentration in agar medium is 150 ppm, 100 ppm, 75 pp
m, 50 ppm and 25 ppm were prepared. One platinum loop was inoculated with each of the above-mentioned bacteria on each of the prepared agar media, and cultured at 35 ° C. for 24 hours. Thereafter, the presence or absence of the occurrence of colonies was examined, and the case where no colonies occurred was defined as the minimum growth inhibitory concentration. Table 2 shows the results.

Culturing at 35 ° C. for 24 hours is a condition under which the bacteria can grow sufficiently in the case of a medium containing no bactericide.

[0052]

[Table 2]

From Table 2, it can be seen that the composition of the present invention exhibits a bactericidal effect on both vancomycin-resistant enterococci and vancomycin-sensitive enterococci when the composition of the present invention is 100 μg / ml or less. In each case, the composition containing a mixture of aloe, green tea, Kumasa and Dokudami required a lower hinokitiol concentration for sterilization than hinokitiol alone. This is probably because the extract such as aloe enhanced the bactericidal effect of hinokitiol.

[0054]

The fungicide for vancomycin-resistant enterococci of the present invention uses hinokitiol, which has low toxicity, as an active ingredient, and is widely used as a fungicide for drinking, spraying, and application in pharmaceuticals, daily necessities, food production sites, and the like. , Available.

By containing an extract such as aloe, the amount of hinokitiol required for sterilization can be reduced.
In addition, in the case of an aqueous bactericide, the solubility and solubility of hinokitiol can be increased to reduce the dosage and usage of the aqueous bactericide.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 35/78 A61K 35/78 BSQHTKE 47/04 47/04 A61L 2/16 A61L 2 / 16 Z 2/18 2/18 A61P 31/04 A61P 31/04

Claims (4)

[Claims] 1. Hinokitiol or a metal complex thereof,
Or a fungicide for vancomycin-resistant enterococci, which comprises a salt thereof as an active ingredient. 2. The fungicide for vancomycin-resistant enterococci according to claim 1, further comprising at least one extract selected from the group consisting of aloe, green tea, Kumasa bamboo and dokudami. 3. Persimmon leaves, pine, cedar, sweet tea, perilla, wasabi, madder, plum, garlic, peppermint,
The sterilizing agent for vancomycin-resistant enterococci according to claim 1 or 2, comprising at least one extract selected from the group consisting of mugwort, salmon, rhubarb, thistle, peppermint, loquat, purple, lavender, lemongrass, and peppermint. Agent. 4. The fungicide for vancomycin-resistant enterococci according to claim 1, which is an aqueous solution containing water as a carrier.