KR20200080804A - Composition comprising grapefruit fermented extract and squalene oil fermented product and preparation method thereof - Google Patents

Abstract

The present invention relates to a composition comprising a fermented grapefruit and a preparing method thereof and, more specifically, to a composition for treating companion animal skin diseases, comprising a fermented product of lactic acid bacteria of an ultra-high pressure liquefied extract of a grapefruit and a microbial fermented product of squalene oil, and a preparing method thereof. The composition for treating companion animal skin diseases, comprising a fermented grapefruit and a microbial fermented product of squalene oil has high skin affinity, does not cause side effects such as allergies, is suitable for long-term use, and can help treat skin diseases caused by underlying inflammation by suppressing inflammatory factors. Also, the composition does not have cytotoxicity, so that degeneration and damage of tissue are small, and it does not cause problems even in ingestion of companion animals, so it can be used in the form of a cosmetic composition or pharmaceutical composition for skin treatment for companion animals.

Description

Composition comprising grapefruit fermented and squalene oil fermented product and its manufacturing method{Composition comprising grapefruit fermented extract and squalene oil fermented product and preparation method thereof}

The present invention relates to a composition comprising a grapefruit fermentation product and a squalene oil fermentation product and a method for manufacturing the same, and more particularly, a companion animal skin disease comprising a lactic acid bacteria fermentation product of grapefruit ultra high pressure liquefied extract and a microorganism fermentation product of squalene oil. It relates to a composition for improvement and a method for manufacturing the same.

As the population structure is aging and the number of single households increases, the number of people who recognize pets as family members or companions is steadily increasing, and the pet market is also steadily growing in various fields such as toys, food, medicines, etc. 2020 It is expected to rapidly expand to 6 trillion won in growth.

The increase in the aging population is contributing to the improvement of the market size of pets, and is putting a lot of time and effort into improving the health of pets, especially in the trend of rapidly spreading pet culture using social network services (SNS) among young people. Demand for medical examination and treatment of pets is increasing.In the past, if drugs for humans were predominant, the interest in diseases of pets is now increasing, so the development and production of pharmaceuticals for pets Increasingly, the interest in skin disease care of companion animals, which are particularly vulnerable to skin diseases, has increased significantly.

The most common cause of visits to domestic animal hospitals is due to skin diseases, which account for about 24%, and about 25% of all companion animals are affected by fungi or bacteria such as eczema, swelling, keratin, folliculitis, ear infections, ringworm, etc. It was investigated that they suffer from skin diseases caused by (PNV provided by Electronic Chart of Animal Hospital, 2015).

Domestic veterinary medicines are rapidly increasing from 3984 billion won in 2006 to 583.7 billion won in 2012, but a large number of medicines are causing side effects such as allergies, skin damage, and weakening of immunity due to excessive use of antibiotics. Disinfectants, Steroids Because of the incense, most of the things give consumers a sense of rejection, so the solution is urgent.

Republic of Korea Registered Patent Publication No. 1593624 (2016.02.15.) Republic of Korea Patent Publication No. 2018-0121654 (2018.11.07.)

An object of the present invention is to provide a composition for improving a skin disease that is safe for a companion animal, has high skin affinity, and does not cause side effects such as allergies, and does not show rejection even in long-term use.

Another object of the present invention is to improve the skin disease, which can show a strong antibacterial and anti-inflammatory action against the fungal or bacterial skin disease of companion animals using natural extracts while preventing the expansion of secondary infections and show a high rate of skin disease improvement. It is to provide a composition.

The method for preparing a composition for improving a skin disease of a companion animal according to the present invention comprises the steps of: a) preparing a grapefruit extract from grapefruit by an ultra-high pressure liquefaction extraction method; b) preparing a grapefruit fermentation product by fermenting the grapefruit extract; And c) fermenting squalene oil to produce a squalene oil fermentation product.

In the method for preparing a composition for improving a skin disease of a companion animal according to an aspect of the present invention, the step a) comprises: a1) preparing grapefruit extract by extracting grapefruit in an ultra-high pressure liquefaction with an extraction solvent; a2) centrifuging the grapefruit extract, followed by filtering to obtain a filtrate; And a3) concentrating and filtrating the filtrate.

In the method for preparing a composition for improving skin diseases of companion animals according to an aspect of the present invention, the ultra-high pressure liquefaction extraction method may be performed under 10 to 1000 MPa.

In the method of preparing a composition for improving skin diseases of companion animals according to an aspect of the present invention, the fermentation of step b) may be performed after adding Lactobacillus pentosus .

In the method for preparing a composition for improving a skin disease in a companion animal according to an aspect of the present invention, the fermentation in step c) is Pseudozyma tsukubaensis ) It can be carried out after adding KCTC7770.

In the method of preparing a composition for improving skin diseases of companion animals according to an aspect of the present invention, the grapefruit fermentation product and the squalene oil fermentation product may be mixed in a weight ratio of 10:1 to 50:1.

The cosmetic composition for improving a companion animal's skin disease according to the present invention includes a composition for improving a companion animal's skin disease prepared by the above manufacturing method.

The pharmaceutical composition for improving a companion animal's skin disease according to the present invention includes a composition for improving a companion animal's skin disease prepared by the above manufacturing method.

In the pharmaceutical composition for improving companion animal skin disease according to an aspect of the present invention, the skin disease may be dermatitis, eczema, folliculitis, pyoderma, ear disease or ringworm.

The composition for improving a companion animal's skin disease comprising a grapefruit fermentation product and a microbial fermentation product of squalene oil according to the present invention has high skin affinity, does not cause side effects such as allergies, is suitable for long-term use, and suppresses inflammation-inducing factors It can help to improve skin diseases caused by underlying inflammation. Also, it has no cytotoxicity, so there is little tissue degeneration and damage, and it does not cause problems with the consumption of pets. It can be widely used in the form of.

1 is a result of confirming the effect of improving the disease after a certain period of time after applying the composition for improving skin diseases of the present invention to a diseased area of a companion animal.

Hereinafter, a composition including a grapefruit fermentation product of the present invention and a microbial fermentation product of squalene oil and a manufacturing method thereof will be described in detail with reference to the accompanying tables or drawings.

When the drawings are described, they are provided as examples in order to sufficiently convey the spirit of the present invention to those skilled in the art. Therefore, the present invention is not limited to the presented drawings and may be embodied in other forms, and the drawings may be exaggerated to clarify the spirit of the present invention.

At this time, unless there are other definitions in the technical terms and scientific terms used, it has the meanings commonly understood by those of ordinary skill in the art to which this invention belongs, and the subject matter of the present invention in the following description and accompanying drawings Descriptions of well-known functions and configurations that may be obscured are omitted.

Also, the singular form used in the specification of the present invention may be intended to include the plural form unless otherwise specified in the context.

In addition, in the specification of the present invention, the units used without specific reference are based on weight, and for example, units of% or ratio mean weight percent or weight ratio.

Also, in the specification of the present invention, the expression “comprises” is an open description having the meaning equivalent to expressions such as “have”, “contains”, “haves” or “makes a feature”, and is further enumerated. It does not exclude elements, materials or processes that are not present. Also, “actually… The phrase “consisting of” means that a specified element, material, or process, along with other elements, materials, or processes not listed, does not have an unacceptably significant effect on at least one basic and novel technical idea of the invention. It means that it can exist in quantity. Also, the expression “consisting of” means that only the element, material, or process described is present.

These researchers have developed ingredients for improving the skin diseases of companion animals that are safe for the companion animal's skin and are safe in the body even when ingested and do not cause side effects such as allergies due to their high skin affinity. While researching to find out, the composition for improving skin diseases using grapefruit fermentation and microbial fermentation of squalene oil shows strong antibacterial and anti-inflammatory action against fungal or bacterial skin diseases of companion animals and prevents secondary infection expansion. The present invention was completed by confirming a high improvement rate for skin diseases.

Hereinafter, a method of manufacturing a composition for improving a companion animal skin disease according to the present invention will be described in detail.

The method for preparing a composition for improving a skin disease of a companion animal according to the present invention comprises the steps of: a) preparing a grapefruit extract from grapefruit by an ultra-high pressure liquefaction extraction method; b) preparing a grapefruit fermentation product by fermenting the grapefruit extract; And c) fermenting squalene oil to produce a squalene oil fermentation product.

In the step a) of producing a grapefruit extract from grapefruit using ultra-high pressure liquefaction extraction, ultra-high pressure liquefaction extraction (HPP) refers to a technique of physically extracting at a very high pressure, and an ultra-high-pressure liquefaction extraction device known in the art Can be used without limitation.

The pressure at the time of extraction is not particularly limited as long as the active ingredient is extractable from grapefruit Robo, for example, 10 to 1000 MPa, preferably 20 to 500 MPa, more preferably 50 to 100 MPa, and In the range, the active ingredient of grapefruit can be extracted at a high concentration, and it is preferable because it does not need to be separated and used.

The extraction temperature in the step a) is not particularly limited as long as the active ingredient can be extracted without deterioration or denaturation, and for example, 70° C. or less, preferably 50° C. or less, more preferably 30° C. or less, best May be 15° C. or less, and the lower limit of the extraction temperature is not limited as long as extraction using an extraction solvent is possible, for example, a temperature higher than the freezing point of the extraction solvent, etc., and a non-limiting example is 5° C. The above can be exemplified. This is performed when the extraction is performed at a high temperature during ultra-high pressure liquefaction extraction, because problems such as destruction of an active ingredient by heat, protein denaturation, and destruction of plant components may occur. It is preferable, and the effect of the active ingredient of the extract containing grapefruit can be remarkably improved.

The extraction time in step a) is not particularly limited as long as the active ingredient can be sufficiently extracted, for example, 0.1 to 48 hours, preferably 1 to 24 hours, more preferably 1.5 to 12 hours, most preferably It may be 2 to 6 hours, it is preferable because there is no problem such as deterioration of the active ingredient extracted in the above range.

The extraction solvent in the step a) is not particularly limited as long as it is a component capable of extracting an active ingredient capable of maintaining a skin disease-absorbing and preserving property from the raw material. It is possible to use an organic solvent such as ethanol, and by applying the extraction method according to the present invention, it is possible to extract sufficient active ingredients even when water is used without using an organic solvent that may exhibit toxicity depending on the ingredients. desirable. However, this is only an example, and the scope of the present invention is not limited thereto.

The mixing ratio of the extraction raw material and the extraction solvent in the step a) is not particularly limited as long as the active ingredient can be sufficiently extracted, for example, 50 to 2000 parts by weight of the extraction solvent per 100 parts by weight of grapefruit, preferably 100 to 1500 It may be parts by weight, more preferably 300 to 1000 parts by weight, and is preferable because extraction is possible without loss of an active ingredient in the above range.

The extract containing the active ingredient obtained through the extraction method as described above may be used as it is for fermentation in step b), but additionally undergoes steps such as filtration or drying as described below for ease of removal of impurities or storage. It is possible to obtain a composition having an improved content of the active ingredient through this, thereby improving skin disease or improving the treatment effect.

The step a) comprises: a1) preparing grapefruit extract by extracting grapefruit with ultra-high pressure liquefaction with an extraction solvent; a2) centrifuging the grapefruit extract, followed by filtering to obtain a filtrate; And a3) concentrating and pulverizing the filtrate.

In step a2), centrifugation may be performed by appropriately introducing a known method, and matters not mentioned in the following descriptions are not limited to those performed by those skilled in the art from the prior art documents.

The centrifugation speed and the centrifugation time are, for example, 100 to 30,000 rpm, preferably 500 to 20,000 rpm, more preferably 1,500 to 15,000 rpm at 10 seconds to 30 minutes, preferably 30 seconds to 20 minutes, more preferably 1 minute to 10 minutes can be performed. However, this is only an example, and the scope of the present invention is not limited thereto.

The filtration is not limited as long as it is a method capable of separating or removing the residual solids after extraction of the active ingredient, for example, dialysis, reverse osmosis (RO), microfiltration , Ultrafiltration, and the like, preferably, ultrafiltration. The molecular weight cut (molecular cutoff) of the filtration membrane during ultrafiltration may be 10 kDa to 1000 kDa, preferably 30 kDa to 500 kDa, more preferably 50 kDa to 200 kDa, and contains less impurities other than active ingredients in the above range, extract It is preferable because it can improve the purity of.

In the step a3), the concentration can be performed by appropriately referring to a conventionally well-known document by a person skilled in the art so that the extract can be prepared in powder form. At this time, concentration and drying are performed for smooth performance of the powdering process. It may include. The drying method may be selected from various methods such as vacuum drying, hot air drying, natural drying, and freeze drying, and vacuum drying may be used as an example, but this is only an example, and the scope of the present invention is It is not limited to this.

In the step b) of fermenting the grapefruit extract to prepare a grapefruit fermentation product, the fermentation may be fermented according to fermentation conditions known in the art for the strain through inoculation of the strain. That is, conditions such as composition, temperature, pH, and fermentation time of the fermentation medium according to the strain may be selected by those skilled in the art according to the type of strain used for fermentation.

The fermentation medium may include a substrate such as glucose, fructose, galactose, rhamnose or mannose, disaccharide, and polypeptone, yeast extract, ammonium sulfate, potassium phosphate, magnesium sulfate, calcium chloride, and the like, but this is one example. It is only, and can be used by appropriately selecting a medium that can be used depending on the object of fermentation, the type of strain or fermentation conditions, and the scope of the present invention is not limited thereto.

The fermentation temperature is not limited as long as the viable temperature of the strain to be added and the temperature at which the active ingredient is not denatured, for example, 20 to 40°C, preferably 22 to 38°C, and more preferably 25 to 35°C. It is possible to improve the skin and improve the effectiveness of the active ingredient through this, which is preferable.

The range of pH in the fermentation tank at the time of fermentation is not limited so long as the viable temperature of the strain to be added and the range in which denaturation of the active ingredient does not occur, for example, pH 5 to pH 10, preferably pH 6 to pH 9, more preferably It may be a pH of 6.5 to 8.5, it is preferable because the effect of improving the skin of the active ingredient, such as improving inflammation can be improved.

The fermentation time during the fermentation is not limited if fermentation is sufficiently progressed to obtain an active ingredient, and may be, for example, 1 to 72 hours, preferably 4 to 48 hours, more preferably 8 to 24 hours, The fermentation time may be adjusted by a person skilled in the art according to the growth curve of the strain.

The type of strain that can be added at the time of fermentation of the grapefruit extract is preferably a strain that can significantly improve the skin disease improvement effect and inflammation improvement effect of the active ingredient through fermentation, and as an example, lactic acid bacteria can be used. have. The lactic acid bacteria include Lactobacillus pentosus , Lactobacillus plantarum strain, Lactobacillus paracasei strain, Pediococcus pentosaceus strain, Lactobacillus strain Lactobacillus casei strain, Lactobacillus sakei strain, Lactobacillus brevis strain, Streptococcus thermophillus strain, Leuconostoc mesenteroid ( Leuconostoc mesenteroides strain, Leuconostoc citreum ) strain and Wissella cibaria strain are preferably performed using at least one selected from the group consisting of strains, more preferably, it can be performed using Lactobacillus pentosus , but are not limited to this. Does not work.

The “lactic acid bacteria” fermentation is preferably performed for 1 to 3 days at a temperature of 30°C to 50°C, and more preferably for 1.5 to 2.5 days at a temperature of 30°C to 40°C, but is not limited thereto. The “lactic acid bacteria” fermentation has an advantage of optimizing pH and total acidity by maintaining the temperature range and the time range.

In the step c) of fermenting squalene oil to prepare a squalene oil fermentation product, the fermentation may be fermented according to fermentation conditions known in the art for the strain through inoculation of the strain. That is, conditions such as composition, temperature, pH, and fermentation time of the fermentation medium according to the strain may be selected by those skilled in the art according to the type of strain used for fermentation.

The fermentation medium may include a substrate such as glucose, fructose, galactose, rhamnose or mannose, disaccharide, and polypeptone, yeast extract, ammonium sulfate, potassium phosphate, magnesium sulfate, calcium chloride, and the like, but this is one example. It is only, and can be used by appropriately selecting a medium that can be used depending on the object of fermentation, the type of strain or fermentation conditions, and the scope of the present invention is not limited thereto.

The fermentation temperature is not limited as long as the viable temperature of the strain to be added and the temperature at which the active ingredient is not denatured, for example, 20 to 40°C, preferably 22 to 38°C, and more preferably 25 to 35°C. It is possible to improve the skin and improve the effectiveness of the active ingredient through this, which is preferable.

The range of the pH in the fermentation tank during the fermentation is not limited as long as the viable temperature of the strain to be added and the range in which denaturation of the active ingredient does not occur, for example, pH 5 to pH 10, preferably pH 6 to pH 9, more preferably It may be a pH of 6.5 to 8.5, it is preferable because the effect of improving the skin of the active ingredient, such as improving inflammation can be improved.

The fermentation time during the fermentation is not limited if fermentation is sufficiently progressed to obtain an active ingredient, and may be, for example, 1 to 72 hours, preferably 4 to 48 hours, more preferably 8 to 24 hours, The fermentation time may be adjusted by a person skilled in the art according to the growth curve of the strain.

The type of strain that can be added at the time of fermentation of squalene oil is preferably a strain that can significantly improve the skin disease improvement effect and inflammation improvement effect of the active ingredient through fermentation, for example, Pseudozyma ) sp. One or more strains belonging to the genus can be selected and used. As a strain of the genus Pseudojima , for example, Pseudozyma tsukubaensis ) KCTC7770 may be used, but is not limited thereto. When using the strain, the improvement effect of skin disease and inflammation can be significantly improved, which is preferable.

Squalene oil is known to play an important role in metabolism in vivo, and when 10% or more squalene component is present in the body or skin, the skin disease improvement effect, inflammation improvement effect, skin gloss and elasticity maintenance, anti-wrinkle, etc. It is desirable because it can help.

Grapefruit fermentation and squalene oil fermentation products obtained through steps b) and c) are in a weight ratio of 10:1 to 50:1, preferably 15:1 to 40:1, more preferably 20:1 to 30:1. It can be mixed, in the above range, the skin disease improving effect of the active ingredient, the effect of improving inflammation is significantly increased, and the protective effect on the skin can also be simultaneously obtained synergistic effect is preferable.

The pharmaceutical composition for improving a companion animal's skin disease according to the present invention includes a composition for improving a companion animal's skin disease prepared by the above manufacturing method.

The cosmetic composition is not limited from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays It can be any one of the selected formulations.

The cosmetic composition according to the present invention may include, as an active ingredient, ingredients commonly used in cosmetics in addition to the above composition, for example, conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, and carriers. It can contain.

On the other hand, the cosmetic composition may be prepared in any formulation conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleaning , Oil, powder foundation, emulsion foundation, wax foundation and spray. However, it is not necessarily limited thereto. More specifically, it can be prepared in the form of flexible lotion, converging lotion, nutrition lotion, nutrition cream, massage cream, essence, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the cosmetic composition is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components. Can be.

Further, when the formulation of the cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additionally chlorofluorohydro Propellants such as carbon, propane/butane or dimethyl ether.

In addition, when the formulation of the cosmetic composition is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, and propylene Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

In addition, when the formulation of the cosmetic composition is a suspension, a liquid diluent such as water, ethanol or propylene glycol as a carrier component, a suspension agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester , Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or trakant, and the like.

In addition, when the formulation of the cosmetic composition is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate as a carrier component , Fatty acid amide ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like.

The pharmaceutical composition for improving a companion animal's skin disease according to the present invention includes a composition for improving a companion animal's skin disease prepared by the above manufacturing method.

The pharmaceutical composition for companion animals may be formulated in the form of a conventional transdermal absorption preparation in the pharmaceutical field by adding a conventional non-toxic and pharmaceutically acceptable carrier and excipient.

The pharmaceutically acceptable carrier that can be used in the pharmaceutical composition for a companion animal is commonly used in formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin , Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and/or mineral oil, etc. , But is not limited thereto.

Excipients that can be used in the pharmaceutical composition for companion animals include sweeteners, binders, solubilizers, solubilizers, wetting agents, emulsifiers, isotonic agents, adsorbents, disintegrants, antioxidants, preservatives, lubricants, fillers, fragrances, etc. The proportions and properties of excipients can be determined by the solubility and chemical properties of the selected tablet, the route of administration chosen, and standard drug practice. Examples of excipients are lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, talc, stearic acid, sterin, magnesium stearate, magnesium aluminum silicate, starch, gelatin, tragacanth rubber, arginic acid, Sodium alginate, methyl cellulose, sodium carboxymethyl cellulose, agar, water, ethanol, polyethylene glycol, polyvinylpyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, and vanilla flavor.

In addition, the pharmaceutical composition for the companion animal may be sterilized, or may further include an auxiliary agent such as a preservative, stabilizer, hydration agent, or emulsifying accelerator, salt and/or buffer for osmotic pressure control, and other therapeutically useful substances. It can also be formulated according to conventional methods of mixing, granulating or coating.

In the pharmaceutical composition for improving a companion animal's skin disease according to an aspect of the present invention, the companion animal's skin disease may be caused by expression of an inflammatory trigger, and may include a bacterial disease or an inflammatory disease. Specifically, examples of the bacterial disease or inflammatory disease include dermatitis, eczema, folliculitis, pyoderma, ear disease, or ringworm, but this is only an example, and the scope of the present invention is not limited thereto.

Examples of the inflammatory inducer may include leukotriene, etc. in companion animals, and can reduce tissue degeneration, tissue degeneration and loss due to cytotoxic components through inhibition of the inflammatory factor, and secondary It is preferable because it can prevent the spread of infection due to contact.

Hereinafter, the contents of the present invention will be described in more detail through examples. The examples are only for explaining the present invention in more detail, and the scope of the present invention is not limited by them.

[Example 1]

Ultra high pressure liquefaction extraction process

A mixture of 1 kg of grapefruit and 1 kg of purified water was added to an ultra high pressure device (TFS-2L, TOYO KOATSU, Japan), and extracted at 75 MPa and 15° C. for 1 hour to obtain an ultra high pressure liquefied extract.

The ultra-high-pressure liquefied extract was centrifuged at 9,000 rpm for 10 minutes at 15°C using a centrifuge (Supra 22K, Hanil, Korea), and the filtrate obtained by ultrafiltration with a molecular weight cut of 10 kDa was concentrated under vacuum at room temperature. Then, it was dried in vacuo to obtain a grapefruit extract powder.

Fermentation process 1

Glucose 20 g/ℓ, Polypeptone (BD Difco) 5 g/ℓ, Yeast Extract (BDDifco) 3 g/ℓ, Ammonium Sulfate 1.5 g/ℓ, Potassium Phosphate 1 g/ℓ, Magnesium Sulfate 0.5 g /L, 10 g/L of grapefruit extract powder obtained from the above was added to a fermentation medium containing 0.15 g/L of calcium chloride to prepare a medium composition containing grapefruit extract.

Thereafter, 30 L of the medium composition containing grapefruit extract was inoculated with Lactobacillus pentosus , and cultured for 72 hours to prepare a grapefruit fermented product.

Fermentation process 2

Glucose 20 g/ℓ, Polypeptone (BD Difco) 5 g/ℓ, Yeast Extract (BDDifco) 3 g/ℓ, Ammonium Sulfate 1.5 g/ℓ, Potassium Phosphate 1 g/ℓ, Magnesium Sulfate 0.5 g /L, 250 g/L of olive-derived squalene oil (Squalene Oil, Natural Cosmetics Laboratory) was added to a fermentation medium containing 0.15 g/L of calcium chloride to prepare a squalene oil-containing medium composition.

Subsequently, Pseudozyma tsukubaensis was added to 30 L of the squalene oil-containing medium composition. After inoculating KCTC7770, cultured for 72 hours to prepare squalene oil fermentation products.

Fermentation mix

The fermentation mixture was prepared by mixing the grapefruit fermentation product and the squalene oil fermentation product prepared in the fermentation process 1 and fermentation process 2, respectively, and dried to prepare a powder.

[Comparative Example 1]

In the fermentation of grapefruit extract in Example 1, except for inoculating E. coli BL21 at the same concentration instead of Lactobacillus pentosus , the rest of the process was the same to prepare a grapefruit fermentation product.

[Comparative Example 2]

When fermenting squalene oil in Example 1, except for inoculating E. coli BL21 at the same concentration instead of Psedozyma tsukubaensis , the rest of the procedure was the same to prepare squalene oil fermented products.

[Test Example 1] Evaluation of inflammation treatment effect

In order to improve the inflammation of the fermentation mixture powder according to the present invention and to confirm the therapeutic effect, skin patch is applied to a cat, a dog with a similar size of inflammation on the skin, for 12 dogs, each of 6 dogs, using a haye's test chamber The test was conducted.

First, the experimental sites of each experimental group were washed clean with ethanol for disinfection, dried, and then the powders of Examples 1, 1, and 2 were added dropwise to the chamber by 20 ug, and then fixed on the test site.

The patch was applied for 24 hours, and after removing the patch, the test sites were marked with a marking pen, and the test sites were observed after 48 hours respectively.

The improvement of inflammation is judged by the extent to which the area of the wound is reduced, so it is very good if more than 50% of the inflammatory area is treated, good when 25% to 50% of the inflammatory area is treated, and 10-25% of the inflammatory area In the case of treatment, it was evaluated as usually, ˜10% of the inflammatory site was bad when treated, and very bad when there was no treatment effect.

Each result is shown in Table 1.

Experimental group Inflammation treatment effect Example 1 Very good (88% reduction) Comparative Example 1 Poor (5% reduction) Comparative Example 2 Very bad (no treatment effect)

From the results of Table 1, it was confirmed that the fermentation mixture powder according to the present invention has a very excellent treatment effect on inflammation of a companion animal, compared to the comparative experimental group, and the comparative experimental group has little or no inflammation treatment effect, or treatment compared to the embodiment It was confirmed that the effect was notably good.

More specifically, in the case of Example 1, the inflammation site was reduced by 88%, indicating that the treatment effect was very good, and thus it was confirmed that it was particularly excellent in treating the inflammatory skin diseases of companion animals.

[Formulation Example 1] Preparation of nutrition cream

3% by weight of propylene glycol, 0.5% by weight of dextrin behenate, and 0.2% by weight of benzoin were mixed and stirred at 80°C based on the total weight of the nutrient cream in purified water.

Then, using an emulsifier, 1% by weight of beeswax, 1.5% by weight of polysorbate 60, 0.5% by weight of sorbitan sesquioleate, 10% by weight of liquid paraffin, 1% by weight of sorbitan stearate, 0.5% by weight of glycerin of lipophilic monostearic acid, 1.5% by weight of stearic acid and 1% by weight of glyceryl stearate were emulsified at 80°C. After the emulsification was completed, the mixture was cooled to 50°C while stirring using a stirrer.

Then, 8% by weight of the fermentation mixture powder of Example 1 was added to cool again to 25°C, and aged at 25°C for 2 days to prepare a nutrient cream. The content of the purified water corresponds to the remaining amount of the nutrition cream.

[Formulation Example 2] Preparation of lotion

3% by weight of glycerin, 2% by weight of butylene glycol, 2% by weight of propylene glycol, and 0.2% by weight of benzoin were added to the reaction vessel in order, dissolved in purified water, and dissolved in polyoxyethylene cured castor oil. 1% by weight was dissolved by heating to 60 ℃.

Then, 5% by weight of the fermentation mixture powder of Example 1, 10% by weight of ethanol, and the remaining amount of purified water were sufficiently stirred, and then aged at 25°C for 2 days to prepare cosmetic lotion.

[Formulation Example 3] Preparation of ointment for treatment of inflammation

Ointments were prepared according to the conventional method with the composition shown in Table 2 below.

ingredient Content (% by weight) Fermentation mixture powder of Example 1 5.0 glycerin 8.0 Butylene glycol 4.0 Floating paraffin 15.0 Beta glucan 7.0 Carbomer 0.1 Caprylac/Capric Triglyceride 3.0 Squalane 1.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Cetearyl alcohol 1.2 Wax 4.0 Purified water Balance

[Formulation Example 4] Preparation of a gel for the treatment of inflammation

Gels were prepared according to a conventional method with the composition shown in Table 3 below.

ingredient Content (% by weight) Fermentation mixture powder of Example 1 1.0 Sodium ethylenediamine acetate 0.05 glycerin 4.0 Carboxyvinyl polymer 0.3 ethanol 5.0 PEG 60 hardened castor oil 0.5 Triethanolamine 0.3 Purified water Balance

The specific parts of the present invention have been described in detail above, and it is clear that for those skilled in the art, these specific techniques are merely implementations, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (9)

a) preparing a grapefruit extract from grapefruit by an ultra-high pressure liquefaction extraction method;
b) preparing a grapefruit fermentation product by fermenting the grapefruit extract; And
c) fermenting squalene oil to prepare a fermented product of squalene oil;
Method for preparing a composition for improving skin diseases of companion animals comprising a. According to claim 1,
Step a) is
a1) preparing grapefruit extract by extracting grapefruit into an ultra-high pressure liquefaction with an extraction solvent;
a2) centrifuging the grapefruit extract, followed by filtering to obtain a filtrate; And
a3) concentrating and pulverizing the filtrate;
Method for preparing a composition for improving skin diseases of companion animals comprising a. According to claim 1,
The ultra-high pressure liquefaction extraction method is to perform under 10 to 1000 MPa, a method for preparing a composition for improving skin diseases of companion animals. According to claim 1,
The fermentation of step b) is performed after adding Lactobacillus pentosus , a method for preparing a composition for improving skin diseases of companion animals. According to claim 1,
The fermentation in step c) is Pseudozyma tsukubaensis ) It is performed after adding KCTC7770, a method for preparing a composition for improving skin diseases of companion animals. According to claim 1,
The grapefruit fermentation product and squalene oil fermentation product are mixed in a weight ratio of 10:1 to 50:1, a method for preparing a composition for improving skin diseases of companion animals. A cosmetic composition for improving a skin disease of a companion animal, comprising a composition for improving a skin disease of a companion animal prepared by the method of any one of claims 1 to 6. A pharmaceutical composition for improving a skin disease of a companion animal, comprising a composition for improving a skin disease of a companion animal prepared by the method of any one of claims 1 to 6. The method of claim 8,
The skin diseases are dermatitis, eczema, folliculitis, pyoderma, ear disease or ringworm pharmaceutical composition for improving companion animal skin diseases.

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