JP2020180100A - Control agent and soil treatment methods - Google Patents

Abstract

To provide more effective control means for Cryptosporidium.SOLUTION: Provided is a control agent for Cryptosporidium, which contains a compound represented by the general formula (I) as an active ingredient. The compound inhibits Cryptosporidium's ability to invade host epithelial cells and has the effect of suppressing the infection. Therefore, it may be possible to effectively control the growth and spread of Cryptosporidium protozoa by spraying it on livestock farms as a control agent for Cryptosporidium. It may also be effective as a prophylactic or therapeutic agent for cryptosporidiosis.SELECTED DRAWING: Figure 1

Description

The present invention relates to a preparation for controlling Cryptosporidium containing an isothiocyanic acid compound or the like as an active ingredient, a soil treatment method for controlling Cryptosporidium using these preparations, a therapeutic agent for Cryptosporidium disease, and the like.

Cryptosporidium is a protozoan belonging to the parasitic protozoa and is widely distributed all over the world. For example, Cryptosporidium parvum (scientific name "Cryptosporidium parvum") has been shown to have a high infection rate as an intestinal parasite in cattle and pigs, and has been reported to be detected in dogs and cats.

Cryptosporidium is an obligate cell-parasitic protozoan and is said to develop in the following life cycle. When Cryptosporidium oocysts are excreted with feces and taken orally into the host, the oocysts decapsulate in the intestine and release four sporozoites in the oocysts. Sporozoites invade epithelial cells in the intestine to form a sac, which reproduces asexually and proliferates. The proliferated sporozoite repeatedly releases extracellularly and invades into the cell, and further proliferates. After that, when it shifts to the sexual reproduction stage, it differentiates into either a male reproductive mother or a female reproductive mother, and their zygotes develop to become oocysts, which are released and excreted from the host body together with feces.

Cryptosporidium oocysts are known to be extremely resistant to chlorine. Therefore, if the water source is contaminated with oocysts released together with livestock feces, etc., the oocysts of cryptosporidium will be mixed in tap water, etc., resulting in cryptosporidium disease with diarrhea as the main symptom in humans. May occur collectively. Cryptosporidium disease in humans usually heals spontaneously in about 1 to 2 weeks, but in the case of patients with immunodeficiency, it may lead to death. In humans, cryptosporidium disease is treated with drugs such as paromomycin and nitazoxanide in severe cases.

Cryptosporidium is widespread in livestock such as cattle and pigs. In particular, calves aged 1 month or younger are easily infected by contamination by oocysts in the breeding environment, show symptoms of watery diarrhea, and may die due to secondary infection, resulting in great economic loss. On the other hand, few effective preventive and therapeutic measures for cryptosporidium disease in calves have been established.

As countermeasures against cryptosporidium, for example, Patent Document 1 describes a cryptosporidiosis control agent containing cashew nut shell oil and the like, and Patent Document 2 describes a treatment for cryptosporidium disease containing an iron compound as an active ingredient. -Preventive agents are listed respectively.

In addition, as a matter related to the present invention, Patent Document 3 discloses a method for preventing / treating hairy wart disease by administering allyl isothiocyanate to the limbs of an ungulate.
JP-A-2015-30717 Japanese Unexamined Patent Publication No. 2010-13408 Japanese Patent No. 5740062

As mentioned above, while Cryptosporidium is widespread, effective control measures are required from the viewpoints of public health and livestock economy. Therefore, an object of the present invention is to provide a highly effective control means for Cryptosporidium.

The present inventors have newly found that an isothiocyanic acid compound such as allyl isothiocyanate significantly suppresses the infectivity of Cryptosporidium.

［式I中、Ｒ_１は炭素原子数2〜8のアルケニル基、炭素原子数1〜6の低級アルキル基、炭素原子数6〜12のアリール基、炭素原子数7〜20のアラルキル基、又は炭素原子数3〜6のシクロアルキル基を表す。］ Therefore, the present invention provides a Cryptosporidium control preparation containing a compound represented by the following general formula (I) as an active ingredient.

[In Formula I, R ₁ is an alkenyl group with 2 to 8 carbon atoms, a lower alkyl group with 1 to 6 carbon atoms, an aryl group with 6 to 12 carbon atoms, an aralkyl group with 7 to 20 carbon atoms, or Represents a cycloalkyl group having 3 to 6 carbon atoms. ]

This compound has the effect of inhibiting the ability of Cryptosporidium to invade host epithelial cells and suppressing its infection. Therefore, for example, as a preparation for controlling Cryptosporidium, by spraying it on livestock breeding farms, for example, in barns such as cattle and pigs, or in the soil in ranches, the growth and spread of Cryptosporidium protozoa and the occurrence of Cryptosporidium disease occur.・ There is a possibility that exacerbation, transmission, and spread can be effectively controlled. It may also be effective as a prophylactic or therapeutic agent for cryptosporidium disease.

INDUSTRIAL APPLICABILITY According to the present invention, the occurrence / exacerbation of cryptosporidium disease and the transmission / spread of the pathogenicity can be effectively suppressed.

<About the Cryptosporidium control preparation according to the present invention>
The present invention includes all Cryptosporidium control preparations containing a compound represented by the following general formula (I) as an active ingredient. Here, "Cryptosporidium control" means control against Cryptosporidium, that is, suppression of the infectivity and proliferation power of Cryptosporidium, and "Cryptosporidium control preparation" is used for control of Cryptosporidium. That is, a preparation for suppressing the infectivity, proliferation power, etc. of Cryptosporidium (the same shall apply hereinafter).

In the above general formula I, R ₁ is an alkenyl group having 2 to 8 carbon atoms, a lower alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 12 carbon atoms, and an aralkyl group having 7 to 20 carbon atoms. Alternatively, it represents a cycloalkyl group having 3 to 6 carbon atoms.

The alkenyl group having 2 to 8 carbon atoms in R ₁ may be a linear or branched chain unsaturated hydrocarbon group having 2 to 8 carbon atoms having at least one double bond. Examples of such a group include a vinyl group, a 1-propenyl group, an allyl group (2-propenyl group), a butenyl group, a pentenyl group, a hexenyl group, a heptenyl group, an octenyl group and the like.

The lower alkyl group having 1 to 6 carbon atoms in R ₁ may be a linear or branched lower alkyl group having 1 to 6 carbon atoms. Such groups include, for example, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group, n-hexyl group and the like. Can be mentioned.

Examples of the aryl group having 6 to 12 carbon atoms in R ₁ include a phenyl group, an o-tolyl group, an m-tolyl group, a p-tolyl group, a xylyl group (including all its positional isomers), and a trimethylphenyl group. Examples thereof include a tetramethylphenyl group (including all the positional isomers), a 1-naphthyl group, a 2-naphthyl group, and a biphenyl group.

The aralkyl group having 7 to 18 carbon atoms in R ₁ may be an aryl C1-C6 alkyl group in which one hydrogen atom of the lower alkyl group is substituted with the aryl group. Such groups include, for example, a benzyl group, a phenethyl group, a phenylpropyl group, a 1-naphthylmethyl group, a 2-naphthylmethyl group, a 1-naphthylethyl group, a 2-naphthylethyl group, a methylnaphthyl group (its positional isomers). Includes all), dimethylnaphthyl group (including all its positional isomers), biphenylmethyl group, biphenylethyl group and the like.

Examples of the cycloalkyl group having 3 to 6 carbon atoms in R ₁ include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group and the like.

Among them, in the formula I, it is preferable that R ₁ is an alkenyl group having 2 to 6 carbon atoms, R ₁ is an allyl group, i.e., the compound is allyl isothiocyanate (Ariruiso Thiocyanate) is most preferred.

As the Cryptosporidium control preparation according to the present invention, known dosage forms such as solid (for example, powder, granule, tablet, etc.) and liquid (for example, liquid, suspending agent, etc.) can be widely adopted. For example, a solid preparation may be dissolved in a solvent at the time of use to prepare a control solution and used, or a liquid preparation may be prepared in advance and, in some cases, diluted appropriately at the time of use for control. It may be used as a solution.

As the solvent for dissolving the compound according to the present invention, a known solvent can be appropriately adopted. For example, an appropriate one may be selected and used from water, vegetable oil, liquid animal oil, phosphate buffer, carbonic acid buffer, physiological saline, alcohol, propylene glycol, polyethylene glycol, methyl cellulose, acetone, DMSO and the like.

In addition, excipients, disintegrants, binders, lubricants, coating agents, stabilizers, solid carriers, solubilizers, buffers, isotonic agents, surfactants, etc., depending on the purpose, application, dosage form, etc. Agents, preservatives, antibacterial agents, antioxidants, pH regulators, dispersants, fragrances, colorants, thickeners and the like may be added as appropriate.

Examples of excipients include sugar alcohols (eg, erythritol, mannitol, xylitol, sorbitol, etc.), sugars (eg, sucrose, powdered sugar, lactose, glucose, etc.), cyclodextrins (eg, α-cyclodextrin, β). -Cyclodextrin, γ-cyclodextrin, hydroxypropyl β-cyclodextrin and sulfobutyl ether β-cyclodextrin sodium, etc., celluloses (eg, crystalline cellulose, microcrystalline cellulose, powdered cellulose, hydroxypropyl cellulose, low degree of substitution hydroxypropyl) Cellulose, ethyl cellulose, methyl cellulose, etc.), starches (eg, corn starch, potato starch, wheat starch, rice starch, partially pregelatinized starch, pregelatinized starch, etc.), dextrin, maltodextrin, sucrose ester, povidone, hypromellose Phtalate, hydroxypropylmethylcellulose acetate succinate, magnesium aluminometasilicate, synthetic aluminum silicate, light anhydrous silicic acid, calcium silicate, sodium carmellose, hypromellose, gum arabic, sodium alginate, gelatin, purulan, polyvinyl alcohol, Examples include carboxyvinyl polymer.

Examples of disintegrants are crospovidone, carmellose, carmellose calcium, croscarmellose sodium, low substitution hydroxypropyl cellulose, sodium starch glycolate, corn starch, potato starch, wheat starch, rice starch, partially pregelatinized starch, α. Examples include modified starch, sodium carboxymethyl starch, agar, and gelatin powder.

Examples of the binder include hydroxypropyl cellulose, carboxymethyl cellulose, ethyl cellulose, methyl cellulose, carmellose sodium, polyvinylpyrrolidone and the like.

Examples of lubricants include stearic acid, magnesium stearate, calcium stearate, talc, hydrous silicon dioxide, light anhydrous silicic acid, sucrose fatty acid esters, hydrogenated vegetable oils, macrogol and the like.

Examples of coating agents include sucrose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, gelatin, glycerin, sorbitol, ethyl cellulose, polyvinylpyrrolidone, cellulose acetate phthalate, hydroxypropyl methyl cellulose phthalate, methyl tetaacrylate / methacrylic acid copolymer, aminoalkyl methacrylate. Examples thereof include copolymer E, aminoalkyl methacrylate copolymer RS, cellulose acetate phthalate, methacrylic acid copolymer L, methacrylic acid copolymer LD, and methacrylic acid copolymer S.

Examples of stabilizers include fatty acids and salts thereof, methylparaben, parahydroxybenzoic acid esters (eg, propylparapen, etc.), alcohols such as chlorobutanol, penzyl alcohol, phenylethyl alcohol, thimerosal, acetic anhydride, sorbic acid, Sodium hydrogen sulfite, L-ascorbic acid, sodium ascorbate, butyl hydroxyanisole, petit hydroxytoluene, propyl gallate, tocopherol acetate, gentidic acid, ascorbic acid, benzyl alcohol, tocopherol, gallic acid, gallic acid ester, α-thioglycerol , Α-tocopherols, proteins and polysaccharides.

Examples of solid carriers include natural mineral powders (eg, cationic clay, bilophilite clay, bentonite, montmorillonite, kaolin, clay, talc, chalk, quartz, attapulsite, diatomaceous earth, etc.), synthetic mineral powders (eg, silicic acid, etc.) Alumina, silicate, etc.), high molecular weight natural products (for example, crystalline cellulose, corn starch, gelatin, alginic acid, etc.) and the like.

Examples of solubilizers include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like.

As an example of the buffer, a buffer solution such as phosphate, acetate, carbonate, citrate tartrate, trishydroxymethylaminomethane, HEPES and the like can be used.

As an example of the tonicity agent, sodium chloride, glucose, D-sorbitol, glycerin, D-mannitol and the like can be used.

Examples of surfactants include nonionic surfactants (eg, aliphatic alcohol ethoxylates, polyoxyethylene alkylphenyl ethers, alkyl glycosides, fatty acid alkanolamides, polyoxyethylene castor oil, hardened castor oil, octyl glucoside, peptylthio. Glucoside, decanoyle-N-methylglucamide, polyoxyethylene dodecyl ether, polyoxyethylene heptamethylhexyl ether, polyoxyethylene isooctylphenylate, polyoxyethylene nonylphenyl ether, polyoxyethylene fatty acid ester, sucrose fatty acid ester, Polyoxyethylene sorbitol ester, etc.), anionic surfactants (eg, sodium dodecyl sulfate, sodium dodecyl sulfonate, sodium dodecylbenzene sulfonate, sodium dodecyl-N-sarcosinate, sodium colate, sodium deoxycholate, tauro Sodium deoxycholate, hydrogenated coconut fatty acid monoglyceride, sodium monosulfate, sodium α-olefin sulfonate, sodium N-palmitoyl glutarmate, sodium N-methyl-N-acyltaurine, etc.), cationic surfactants (eg, sodium acyltaurine) Lauryl trimethylammonium chloride, stearyltrimethylammonium chloride, benzethonium chloride, cetylpyridinium chloride, benzalconium chloride, stearyldimethylbenzylammonium chloride, cetyltrimethylammonium bromide, tetradecylammonium bromide, dodecylpyridinium chloride, etc.), amphoteric surfactant (palm) Oil fatty acids Amidopropyl betaine, lauryl dimethylaminoacetic acid betaine, lauryl dimethyl amine oxide, 2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazolium betaine, N-lauryl diaminoethyl glycine, N-myristyl diaminoethyl glycine, N- Alkyl-1-hydroxyethyl imidazoline betaine sodium, 3-[(3-coramideppil) dimethylammonio] -1-propanesulfonic acid, 3-[(3-colamidpropyl) dimethylammonio] -2-hydroxy-1- Propane sulfonic acid, palmitoyl lysolecithin, dodecyl-N-betaine, dodecyl-β-alanine, etc.) and the like.

Examples of preservatives include parabens (eg, sodium benzoate, methylparaben, ethylparaben, propylparaben, butylparaben, etc.), cetylpyridinium chloride, alkyldiaminoethylglycine hydrochloride, potassium sorbate, thimerosal, paraoxybenzoic acid esters, etc. Methyl paraoxybenzoate, phenoxyethanol, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, and the like can be used.

Examples of antibacterial agents are quaternary ammonium salt-based bactericidal control agents (eg, benzalkonium chloride, benzethonium chloride, etc.), phenol-based bactericidal control agents (eg, 3-methyl-4-isopropylphenol, thymol), chlorohexidine. , Triclosan, cetylpyridinium chloride, zinc gluconate, zinc citrate, antibacterial plant extracts and essential oils (eg, thyme, lemongrass, citrus, lemon, orange, anis, cloves, aniseed, pine, cinnamon, geranium , Rose, mint, lavender, citronella, eucalyptus, peppermint, thymolium, ajowan, byakudan, rosemary, bear vine, freegrass, lemongrass, ratania, Himalayan cedar, etc.).

As an example of the antioxidant, sulfites, ascorbic acid and the like can be used.

Examples of pH adjusters include acids such as hydrochloric acid, carbonate, acetic acid, citric acid, phosphoric acid, boric acid and sulfuric acid, alkali metal hydroxides such as sodium hydroxide, potassium hydroxide, calcium hydroxide and magnesium hydroxide. Alkali metal carbonates or hydrogen carbonates such as sodium carbonate, alkali metal acetates such as sodium acetate, alkali metal citrates such as sodium citrate, bases such as tromethamole, monoethanolamine, diisopropanolamine, etc. may be used. it can.

As an example of the dispersant, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, polyvinylpyrrolidone, polysorbate 80 (TWEEN80) and the like can be used.

Examples of fragrances are fragrance ingredients (eg 1-carboxylic, synamic aldehyde, orange oil, methyl salicylate, eugenol, menthyl acetate, spirantol, ethyl acetate, ethyl butyrate, isoamyl acetate, hexanal, hexenal, methyl anthranilate. , Ethylmethylphenylglycidate, benzaldehyde, vanillin, ethyl vanillin, flaneol, maltol, ethyl maltol, gamma / delta decalactone, gamma / deltown decalactone, N-ethyl-p-menthan-3-carboxamide, menthyl lactate, ethylene Glycol-1-mentyl carbonate, etc.), natural essential oils (eg, winter green oil, clove oil, thyme oil, sage oil, cardamon oil, rosemary oil, majorum oil, nutmeg oil, lavender oil, paracress oil, etc.), fruit-based Flavors (eg lemon, orange, grapefruit, apple, banana, strawberry, blueberry, melon, peach, pineapple, grape, muscat, cherry, squash, etc.), peppermint, sparemint, menthol, yogurt, coffee, brandy, etc. can be used. it can.

Examples of colorants are caramel pigment, cutinashi pigment, anthocyanin pigment, anato pigment, paprika pigment, red flower pigment, red koji pigment, carotene pigment, carotinoid pigment, flavonoid pigment, cochineal pigment, amaranth (red No. 2), erythrosin (red). 3), Allura Red AC (Red No. 40), Ponceau (Red No. 102), Floxin (Red No. 104), Rose Bengal (Red No. 105), Acid Red (Red No. 106), Tartrazine (Yellow No. 4) , Sunset Yellow FCF (Yellow No. 5), Fast Green FCF (Green No. 3), Brilliant Blue FCF (Blue No. 1), Indigo Carmine (Blue No. 2), Blue No. 201, Blue No. 204, Copper Chlorofil, Copper Chlorophyllin Sodium, mica titanium, titanium oxide and the like can be used.

Examples of thickeners include sodium alginate, propylene glycol alginate, locust bean gum, guar gum, sodium caseinate, egg albumin, gelatin agar, starch, chemical starch, carboxymethylcellulose calcium, carboxymethylcellulose sodium, carboxyvinyl polymer, methylcellulose, ethylcellulose. , Hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, carmellose sodium, sodium polyacrylate, polyvinylpyrrolidone, hypromerose, sodium hyaluronate, sodium chondroitin sulfate, sugar alcohol (sorbitol, xylitol, martitol, lactitol, etc.), polyvalent Alcohol (glycerin, propylene glycol, polyethylene glycol, polyvinyl alcohol, etc.), polyvinylpyrrolidone, sodium starch glycolate, sodium starch phosphate, xanthan gum, caragenin gum, tragacant gum, quince seed extract, sodium chondroitin sulfate and the like can be used.

The above-mentioned compound according to the present invention has an infection-suppressing effect on, for example, Cryptosporidium in general, for example, Cryptosporidium parvum.

By using the Cryptosporidium control preparation according to the present invention in a control target object or a control target area, the infectivity of Cryptosporidium present therein can be suppressed and Cryptosporidium can be effectively controlled.

In principle, all objects, places, and areas contaminated with or may be contaminated with Cryptosporidium can be controlled, and are not particularly limited. For example, livestock breeding facilities (livestock facilities such as livestock barns and livestock fertilization facilities, grazing farms, etc.), animal breeding facilities / areas including laboratory animals / domestic animals, water and sewage treatment facilities, reservoirs, medical / nursing facilities / areas, etc. For the control of cryptospolidium such as floors, walls, entrances and exits of meat production / processing facilities, food production / processing facilities, items installed / placed in those facilities, and equipment / equipment used in the work performed in those facilities. The present invention can be applied. It is also effective in controlling Cryptosporidium contamination in liquids such as water and sewage, polluted water, and stored water.

For example, a liquid preparation according to the present invention is prepared in advance or at the time of use, the object to be controlled is immersed in the control solution, or the object to be controlled or the area to be controlled is wiped with a cloth material containing the control solution. Cryptospolidium may be controlled by pest control, the control solution may be sprayed on the object to be controlled or the area to be controlled, sprayed in a mist form with a sprayer, or sprayed in a foam form with a foam nozzle or the like. Cryptospolidium may be controlled by such means. Further, when the control target is a liquid, for example, Cryptosporidium may be controlled by adding the above compound to the liquid and containing the above compound in the liquid. In addition, for example, the powdered preparation may be sprayed / sprayed as it is on the control target object or the control target area.

For example, when controlling Cryptosporidium in a wide area such as a livestock farm, the control solution or powder formulation according to the present invention may be sprayed on the control target area. When controlling cryptosporidium on the floor / wall surface of each facility or objects installed / placed in those facilities, wipe the object / area to be controlled with a cloth material containing the control solution according to the present invention. Alternatively, the control solution or powder formulation according to the present invention may be sprayed substantially uniformly in the form of a mist on the object / area to be controlled with a sprayer or the like, or may be sprayed in the form of bubbles with a foam nozzle or the like. In addition, Cryptosporidium may be controlled at the feet of people entering and exiting the facility by preparing a stepping tank at the entrance and exit of each facility and putting the control solution according to the present invention in the tank. In addition, when controlling cryptosporidium such as instruments and equipment used in the work performed in the facility, those instruments and equipment may be immersed in the control solution according to the present invention, and those instruments and equipment may be immersed. The control solution or powder formulation according to the present invention may be sprayed substantially uniformly in the form of a mist, or may be sprayed in the form of foam using a foam nozzle or the like.

When the cryptospolidium control preparation according to the present invention is used as a liquid preparation or prepared in a liquid state, the concentration of the compound according to the present invention in the control solution is, for example, the final concentration (concentration at the time of use) of 1. ~ 50,000 mg / L is preferred, 10-20,000 mg / L is more preferred, 100-10,000 mg / L is even more preferred, at 150-5,000 mg / L. It is more preferable to have it, and it is most preferable to have 200 to 1,000 mg / L. When the Cryptosporidium control preparation according to the present invention is used as a solid preparation, it is preferable that the preparation contains 1 to 100% by weight of the compound according to the present invention, and 3 to 50% by weight is contained. It is more preferable that the content is 5 to 30% by weight, and it is most preferable that the content is 5 to 30% by weight.

The preparation for controlling Cryptosporidium according to the present invention may contain at least the compound according to the present invention in a predetermined concentration range, and components other than the compound effective for controlling Cryptosporidium and / or other than Cryptosporidium. It may further contain an ingredient effective in controlling the pathogen.

<About the soil treatment method according to the present invention>
The present invention includes all soil treatment methods for controlling Cryptosporidium using the above-mentioned control preparations.

For example, by spraying a control preparation containing a compound represented by the general formula (I) on the entire soil of a livestock farm, Cryptosporidium contamination in the livestock farm can be effectively suppressed.

As the spraying means, known means can be widely adopted and is not particularly limited. For example, Cryptosporidium may be controlled by preparing a liquid preparation according to the present invention in advance or at the time of use and spraying the control solution on the control target area. Further, for example, a powdery preparation may be prepared and sprayed as it is on the control target area.

<Preventive or therapeutic agent for cryptosporidium disease according to the present invention>
The present invention includes all prophylactic or therapeutic agents for cryptosporidium disease containing the above-mentioned compounds as active ingredients.

As described above, this compound has the effect of inhibiting the ability of Cryptosporidium to invade host epithelial cells and suppressing its infection, and as a preventive or therapeutic agent for Cryptosporidium disease in humans, cattle, pigs and the like. May also be effective.

As the prophylactic or therapeutic agent for cryptosporidiosis according to the present invention, known dosage forms such as solid (for example, powder, granule, tablet, etc.) and liquid (for example, liquid, suspending agent, etc.) can be widely adopted. ..

For example, in the case of solid preparations such as powders, granules and tablets, depending on the purpose, application, dosage form, etc., this agent may be added to excipients, disintegrants, binders, lubricants, coating agents, stabilizers, etc. Solid carriers, buffers, tonicity agents, surfactants, preservatives, antibacterial agents, antioxidants, pH regulators, dispersants, fragrances, colorants, thickeners, soothing agents, etc. are appropriately added. May be. As an example of the soothing agent, for example, benzyl alcohol can be used. For each of the other components, for example, the above-mentioned ones can be widely adopted.

For example, in the case of liquid preparations such as liquids and suspending agents, in addition to solvents, dissolution aids, buffers, isotonic agents, surfactants, preservatives, etc., depending on the purpose, use, dosage form, etc. , Antibacterial agent, antioxidant, pH adjuster, colorant, thickener, soothing agent and the like may be appropriately blended. For each component, for example, the above-mentioned ones can be widely adopted.

The agent according to the present invention may be applied to mammals such as humans, livestock (for example, cows and pigs), dogs and cats.

The administration route is not particularly limited as long as the drug reaches the intestine of the target (patient / patient). For example, a solid preparation or a liquid preparation may be orally administered, or may be parenterally administered as an injection, a suppository, a pellet or the like. Further, when applied to cattle, pigs, dogs, cats, etc., it may be contained in food or the like and administered by ingesting it.

The dose may be appropriately determined according to the application target, symptoms, administration method, etc., and is not particularly limited. The dose is, for example, preferably 0.01 to 200 mg / kg (body weight) / day, more preferably 0.05 to 100 mg / kg (body weight) / day, and most preferably 0.1 to 30 mg / kg. (Weight) / day.

In Example 1, it was examined whether the invasion of Cryptosporidium into host intestinal cells was suppressed by allyl isothiocyanate.

HCT-8 cells (human colon cancer-derived cells) were prepared as host cells. HCT-8 cells were seeded on a 96-well plate and cultured to reach a confluent state.

Cryptosporidium was prepared as follows. Cryptosporidium oocysts were accumulated by centrifugation, taurocholic acid and trypsin were added, and the mixture was allowed to stand for 60 minutes at 37 ° C. to decapsulate, and the obtained sporozoites were collected and suspended to use as test materials.

As a test substance, Wasaolo (registered trademark, manufactured by Mitsubishi Chemical Foods Co., Ltd., powder preparation containing 10% by weight of allyl isothiocyanate, the same applies hereinafter) was dissolved in purified water and used.

To each well in which HCT-8 cells were seeded, sporozoite (2 × 10 ⁴ cells / well) and 100 μL of a medium containing allyl isothiocyanate were added, and the mixture was allowed to stand for 48 hours. The medium is a normal medium in which 10% FCS or the like is added to RPMI-1640, and the final concentrations of allyl isothiocyanate are 0.001 μg / mL, 0.01 μg / mL, 0.1 μg / mL, and 1.0 μg / mL, respectively. Wasaolo solution was added so as to be. In addition, in the negative control, the wasaolo solution was not added to the medium, and in the positive control, paromomycin was added to the medium instead of the wasaolo solution so that the final concentration was 1.0 μg / mL.

After allowing to stand for 48 hours in the presence of allyl isothiocyanate, the sporozoites (sporozoites that did not invade HCT-8 cells) remaining in the medium were removed together with the medium, washed with PBS, and then the HCT-8 cells on the bottom surface were removed. Methanol was fixed. Then, immunostaining was performed using the anti-Cryptosporidium antibody as the primary antibody, and the number of Cryptosporidium invading into HCT-8 cells was measured under a microscope.

The results are shown in Figure 1. FIG. 1 is a graph showing the invasion rate of Cryptosporidium into HCT-8 cells in the presence of allyl isothiocyanate. The vertical axis in the figure, "Invasion rate into host cells," is a bar graph of the invasion rate of Cryptosporidium when the number of Cryptosporidium invasion into HCT-8 cells in the absence of allyl isothiocyanate is 100%. Indicates that there is. In the figure, the bar graph of "N" is the negative control (invasion rate of Cryptosporidium into HCT-8 cells in the absence of allyl isothiocyanate, that is, 100%), and the bar graph of "P" is the positive control (paromomycin 1.0 μg /). The invasion rate of Cryptosporidium into HCT-8 cells in the presence of mL) is shown. The bar graphs of "1 to 4" in the figure show HCT-8 cells when allyl isothiocyanate was added at final concentrations of 0.001 μg / mL, 0.01 μg / mL, 0.1 μg / mL, or 1.0 μg / mL, respectively. Represents the invasion rate of Cryptosporidium into.

As shown in FIG. 1, the invasion rate of the protozoan into the host cell was remarkable and concentrated in the presence of allyl isothiocyanate as compared with the invasion rate of Cryptosporidium into the host cell in the absence of allyl isothiocyanate (100%). Dependently decreased. In addition, the invasion of protozoans into host cells was significantly suppressed in the presence of allyl isothiocyanate as compared with the positive control (in the presence of paromomycin). From this result, it was shown that allyl isothiocyanate has an effect of inhibiting the ability of Cryptosporidium to invade host epithelial cells and suppressing the infection.

In Example 1, the graph which shows the invasion rate of Cryptosporidium into HCT-8 cells in the presence of allyl isothiocyanate.

Claims (6)

A Cryptosporidium control preparation containing a compound represented by the following general formula (I) as an active ingredient.

[In Formula I, R ₁ is an alkenyl group with 2 to 8 carbon atoms, a lower alkyl group with 1 to 6 carbon atoms, an aryl group with 6 to 12 carbon atoms, an aralkyl group with 7 to 20 carbon atoms, or Represents a cycloalkyl group having 3 to 6 carbon atoms. ] The preparation for controlling Cryptosporidium according to claim ₁ , wherein R ₁ is an alkenyl group having 2 to 6 carbon atoms in the formula I. The preparation for controlling Cryptosporidium according to claim 1 or 2, wherein the compound is allyl isothiocyanate. The Cryptosporidium control preparation according to any one of claims 1 to 3, which is used for control in livestock farms. A prophylactic or therapeutic agent for cryptosporidium disease containing a compound represented by the following general formula (I) as an active ingredient.

[In Formula I, R ₁ is an alkenyl group with 2 to 8 carbon atoms, a lower alkyl group with 1 to 6 carbon atoms, an aryl group with 6 to 12 carbon atoms, an aralkyl group with 7 to 20 carbon atoms, or Represents a cycloalkyl group having 3 to 6 carbon atoms. ] A soil treatment method for controlling Cryptosporidium using the control preparation according to any one of claims 1 to 4.

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