WO2023146206A1 - Capsule comprising extract of streptococcus infantis strain, and fiber, bedding, and clothing using same - Google Patents

Abstract

Provided are a capsule having an oil extract of a Streptococcus infantis strain impregnated therein, a fiber having the capsule attached thereto, bedding or clothing comprising the fiber, and respective manufacturing methods therefor. According to the method, skin functional material of the Streptococcus infantis strain may be impregnated in the capsule. Thus, the capsule may be attached to a fiber so as to enable manufacturing a skin functional fiber having anti-inflammatory, anti-atopic, itching inhibiting efficacy, and bedding or clothing using the fiber.

Description

Capsules containing extracts of Streptococcus infantis strains, and textiles, bedding and clothing using the same

It relates to a fiber attached with an organic capsule carrying a skin microbiome having beneficial effects on the skin, bedding and clothing using the same, and a manufacturing method thereof.

Microbiome is a general term for various microorganisms such as bacteria and viruses living in the human body, which change in various ways over time and affect human health. The human microbiome accounts for about 1-3% of the human body weight, and 95% of all microorganisms exist in the digestive tract including the large intestine, and are widely distributed in the respiratory tract, oral cavity, skin, and genitals.

Cosmetic fibers (cosmetic functional fibers) are officially defined by the European Committee for Standardization (CEN). Cosmetic fiber is a fiber product containing a substance capable of continuously releasing active ingredients of cosmetic function into the human body, especially the skin. After the active ingredient is supported in the form of a capsule, a binder is used as a post-processing technology to fix the capsule to the surface of the fiber, or by mixing the capsule when manufacturing yarn, the active ingredient is delivered to the skin while using a product using cosmetic fibers. and infiltrate.

On the other hand, in the case of skin microbiome, it is known that there are effects such as moisturizing, whitening, and anti-aging, and due to these effects, it is in the spotlight as an effective substance in cosmetics.

In order to show optimal efficacy using the skin microbiome having such excellent efficacy, it is good to maintain the balance of the microbiome residing in the skin by continuously exposing the skin microbiome effective material to the skin. However, in the case of cosmetics, there is no way to continuously supply effective substances other than continuously using cosmetics at regular intervals, so it is generally not effective to maintain maximum durability by using an excessive amount of ingredients more than necessary for a single use on the skin. I have a problem that I do not have.

Accordingly, there is a need to develop functional textile products capable of continuously releasing the active ingredients of the skin microbiome for a long period of time.

One aspect is to provide a method for preparing a capsule containing an oil extract of a strain of Streptococcus infantis .

Another aspect is to provide a capsule loaded with an oil extract of a Streptococcus infantis strain.

Another aspect is to provide a fiber to which the capsule according to the above aspect is attached.

Another aspect provides bedding or clothing comprising the fiber according to the one aspect.

One aspect provides a method for preparing a capsule containing an oil extract of a strain of Streptococcus infantis .

The method,

(1) using oil as an extraction solvent to prepare an oil extract of Streptococcus infantis strain; and

(2) preparing a capsule containing the oil extract.

The term "extract" includes all materials obtained by extracting components of a strain, regardless of extraction method, extraction solvent, extraction conditions, extracted components or extract form, and is processed or treated by other methods after extracting components of the strain. It also includes materials that can be obtained by For example, the processing or treatment may include dilution, concentration, drying, purification, fractionation, filtration, fermentation, enzymatic treatment, and the like. Therefore, the extract may include an extract, a dilution or concentrate of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof, or a fraction obtained by fractionating the same.

In the step (1), the extraction solvent may be a solvent capable of dissolving oil-soluble components, for example, oil. By using oil as the extraction solvent, oil-soluble components can be extracted from the strain in the form of oil. "Oil extract of a strain" is an extract obtained by extracting useful components of a strain using oil as an extraction solvent. While it is difficult to encapsulate a hydrophilic substance such as a culture medium of a strain inside a capsule having a hydrophobic outer wall, an oil extract of a strain may be supported inside the capsule. The oil extract of the strain supported in the capsule is slowly released to the outside of the capsule and can be continuously delivered and penetrated into the skin over a long period of time.

In the step (1), the oil is not limited to a specific type, but may be, for example, vegetable oil. The above oils are hazelnut seed oil, jadeite oil, red pepper oil, pine leaf oil, green bean coffee oil, marigold flower oil, green tea seed oil, damask rose flower oil, sweet scented oil, evening primrose oil, sugar pumpkin seed oil, stone oil, camellia oil, drumstick seed oil, lavender oil, lime oil, lemon peel oil, lemon oil, rosemary leaf oil, rosewood oil, rosehip oil, marula seed oil, marjoram leaf oil, macadamia nut oil, mandarin leaf oil, meadow foam Seed oil, cottonseed oil, babassu seed oil, basil oil, bergamot oil, borage oil, balsam flower oil, brazil nut oil, bija oil, apple oil, sea buckthorn oil, apricot seed oil, hempseed oil, ginger oil, Scented Geranium Flower Oil, Sweet Almond Oil, Rice Bran Oil, Annatto Seed Oil, Argan Kernel Oil, Avocado Oil, Apricot Kernel Oil, Andiroba Seed Oil, Angelica Root Oil, Angola Seed Oil, Orange Oil, Corn Oil, Olive Oil , wild geranium oil, tangerine peel oil, citron oil, rapeseed oil, ite palm oil, ylang ylang oil, safflower seed oil, inca peanut seed oil, carrot oil, pine nut oil, jasmine oil, carrot oil, azalea flower oil, Sesame Oil, Calendula Oil, Caviar Oil, Soybean Oil, Queensland Nut Oil, Clove Leaf Oil, Tea Tree Oil, Peppermint Oil, Cypress Oil, Grape Seed Oil, Castor Seed Oil, Pistachio Oil, Hazelnut Oil, Hodong Seed Oil, Pumpkin Seed Oil, It may be at least one selected from jojoba oil, red ginseng oil, and white patterned thistle seed oil. The oil may be at least one selected from jojoba oil, macadamia nut oil, and green tea seed oil. The oil may be jojoba oil.

Alternatively, in the step (1), the oil may be a conventional oil used as a cosmetic. The oil may be an oil that is mild to the skin.

The extraction conditions refer to conditions for extracting components of the strain, such as extraction time and extraction temperature. The extraction time is 30 minutes to 120 hours, 30 minutes to 100 hours, 30 minutes to 80 hours, 30 minutes to 60 hours, 2 hours to 120 hours, 2 hours to 100 hours, 2 hours to 80 hours, 2 hours to 60 hours time, 10 hours to 120 hours, 10 hours to 100 hours, 10 hours to 80 hours, or 10 hours to 60 hours, but may be appropriately selected according to other conditions such as extraction solvent and extraction temperature. The extraction temperature may be 10 to 100 ° C, 10 to 80 ° C, 20 to 100 ° C, 20 to 80 ° C, 20 to 60 ° C, 20 to 40 ° C, or room temperature, but depending on other conditions such as extraction solvent and extraction time can be selected appropriately.

In the step (1), the strain is not limited as long as it belongs to the Streptococcus infantis species. For example, the strain may be a Streptococcus infantis CX-4 strain deposited under accession number KCCM12642P.

The strain is about 95% or more, about 96% or more, about 97% or more, about 98% or more, about 99% or more of the 16S rRNA sequence of Streptococcus infantis CX-4 strain (Accession Number: KCCM12642P) , 16S rRNA having about 99.5% or more, or about 99.9% or more sequence identity. The strain may include a 16S rRNA sequence of Streptococcus infantis CX-4 strain (Accession Number: KCCM12642P). The strain comprises a 16S rRNA having at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% sequence identity with SEQ ID NO: 1 it may be The strain may contain the 16S rRNA of SEQ ID NO: 1.

The term "sequence identity" refers to the degree of identity of amino acid residues or bases between sequences after aligning both sequences to maximize matching in a specific comparison region. Sequence identity can be confirmed according to methods known in the art. The percent of sequence identity can be determined using a known sequence comparison program, examples of which include BLASTN (NCBI), CLC Main Workbench (CLC bio), MegAlign™ (DNASTAR Inc), and the like.

Streptococcus infantis strain oil extract has a skin improving effect. Specifically, the skin improvement may include skin condition improvement, skin beauty improvement, and prevention, improvement, or treatment of skin diseases. The skin improvement may be any one or more of anti-inflammatory, anti-atopic, and itching inhibition or alleviation.

The term "anti-inflammatory" may include suppression, amelioration, or alleviation of inflammation, and may refer to any action that eliminates inflammation or inhibits inflammation by participating in the process.

The term “anti-atopic dermatitis” may include suppression, amelioration, or alleviation of atopic dermatitis, and may refer to any action that eliminates atopic dermatitis, alleviates symptoms, or suppresses the onset of atopic dermatitis.

The term "inhibiting or alleviating itching (pruritus)" can refer to any action that inhibits or relieves an unpleasant sensation that causes the urge to scratch or rub the skin.

The term “skin disorder” may include, but is not limited to skin inflammation, atopic dermatitis, itching (pruritus), and the like. The skin disease may include a disease caused by increased expression of IL-1α or TSLP.

The term "prevention" includes inhibiting the development of a disease. The term "treatment" includes inhibition of the development of, alleviation of, or elimination of a disease. The term "improvement" can mean any action that at least reduces a parameter associated with alleviation or treatment of a condition, eg, the severity of a symptom.

The oil extract of the strain of Streptococcus infantis can suppress or reduce the expression of inflammatory cytokines (eg, IL-1α), or atopy and pruritus inhibitory factors (eg, TSLP).

Alternatively, other strains having a skin improving effect may be used instead of the Streptococcus infantis strain. Therefore, another aspect provides a method for preparing a capsule loaded with an oil extract of a strain having a skin improving effect.

In addition, other strains having a skin improving effect may be used together with the Streptococcus infantis strain. Therefore, another aspect provides a method for preparing a capsule loaded with an oil extract of a Streptococcus infantis strain and other strains having a skin improving effect.

The strain having the skin improvement effect may have anti-inflammatory, anti-atopic, and anti-itching or alleviating effects. In addition, the strain having the skin improvement effect may have other types of skin improvement effect in addition to anti-inflammatory, anti-atopic, and inhibition or relief of itching. For example, the skin improvement effect may include wrinkle improvement, moisturizing, skin barrier enhancement, whitening, and the like, but is not limited thereto.

The strain having the skin improvement effect is not limited in its kind as long as it is a strain known in the art. Examples of strains having the skin improvement effect include strains of the genus Streptococcus, strains of the genus Staphylococcus, strains of the genus Epidermidibacterium, genus Micrococcus strains, strains of the genus Bacillus, strains of the genus Lactobacillus, strains of the genus Bifidobacterium, Cutibacterium genus strains, Rhodobatter genus strains, Rotia genus strains, Cocuria genus strains, Corynebacterium genus strains, Pantoea genus strains, Deinococcus genus strains, Weissella genus strains, Goldonia genus strains, Dietgia genus strains, Brevibacillus genus strains, Novosphingobium genus strains, Imella genus strains, Burkholderia genus strains, Photobacteria genus strains, Microbacterium genus strains, etc., but are not limited thereto. Specific strains belonging to each genus may include all strains known in the art having skin improvement efficacy.

In the step (2),

(2-1) adding the oil extract to a solution containing a surfactant and homogenizing to prepare a first solution;

(2-2) preparing a second solution containing a hydrophobic outer wall precursor; and

(2-3) preparing a capsule by mixing and cross-linking the first solution and the second solution.

In the step (2-1), the first solution may be an emulsion.

In the step (2-1), the surfactant may be at least one selected from the group consisting of sodium dodecyl sulfate, sodium dodecylbenzenesulfonate, polyglycol ether, styrene-maleic anhydride copolymer and stearyl methacrylate there is. The surfactant may be one selected from the group consisting of sodium dodecyl sulfate, sodium dodecylbenzenesulfonate, polyglycol ether, styrene-maleic anhydride copolymer, and stearyl methacrylate. The surfactant may be a polyglycol ether.

In the above step (2-1), the particle diameter of the prepared capsule may vary depending on the type of surfactant. When preparing nanocapsules, it is preferable to use a surfactant such as sodium dodecyl sulfate as a surfactant. When preparing microcapsules, it is preferable to use a nonionic surfactant such as polyglycol ether as a surfactant.

In the step (2-1), the surfactant is 0.1 to 10% by weight, 0.1 to 8% by weight, 0.1 to 5% by weight, 0.5 to 10% by weight, 0.5 to 8% by weight based on the total weight of the first solution (emulsion). 0.5 to 5% by weight, 1 to 10% by weight, 1 to 8% by weight, 1 to 5% by weight, 2 to 10% by weight, 2 to 8% by weight, 2 to 6% by weight, 2 to 4% by weight , 5 to 10% by weight, 5 to 8% by weight, or 5 to 7% by weight. If the content of the surfactant is less than the above range, the emulsifying power may be weak and thus capsules may not be prepared. When the content of the surfactant exceeds the above range, it is difficult to proceed with emulsification due to an increase in viscosity due to an excessive amount of emulsifier, and a problem in that the average particle size of the capsule may rather increase due to a decrease in emulsifying power due to high viscosity may occur.

In the above step (2-1), the particle size of the prepared capsule may vary depending on the content of the surfactant. When preparing nanocapsules, the content of the surfactant may be selected within the range of 5 to 10% by weight. When preparing microcapsules, the content of the surfactant can be selected within the range of 0.1 to 5% by weight.

In the above step (2-1), any solution containing a surfactant can be used without limitation as long as it can dissolve the surfactant as a solvent. For example, the solution containing the surfactant may use a solvent capable of dissolving sodium dodecyl sulfate, sodium dodecylbenzenesulfonate, polyglycol ether, styrene-maleic anhydride copolymer or stearyl methacrylate as a solvent. can

In the step (2-1), the oil extract is an oil extract of the strain. Accordingly, details of the oil extract are as described above.

In the step (2-1), the first solution may further contain an additional active ingredient in addition to the oil extract of the strain. For example, the additional active ingredient may be a skin functional material, but is not limited thereto. As long as the skin functional material is known, it may be used without limitation.

In the step (2-1), homogenization may be performed at 500 to 5000 rpm, 500 to 4000 rpm, 500 to 3000 rpm, 1000 to 5000 rpm, 1000 to 4000 rpm, or 1000 to 3000 rpm. When the above rpm range is selected, microcapsules can be prepared.

Alternatively, in the above step (2-1), homogenization may be performed at 5000 to 10000 rpm, 5000 to 9000 rpm, 5000 to 8000 rpm, 5000 to 7000 rpm, 7000 to 10000 rpm, or 7000 to 9000 rpm . When the above rpm range is selected, nanocapsules can be prepared.

In the step (2-1), homogenization is performed for 30 seconds to 1 hour, 30 seconds to 30 minutes, 30 seconds to 20 minutes, 30 seconds to 10 minutes, 1 minute to 1 hour, 1 minute to 30 minutes, 1 minute to 20 minutes, 1 minute to 10 minutes, 5 minutes to 1 hour, 5 minutes to 30 minutes, 5 minutes to 20 minutes, or 5 minutes to 10 minutes.

In the above step (2-1), the particle diameter of the capsule may be changed by adjusting the homogenization rate. The higher the homogenization rate, the smaller the particle size of the capsules. For example, to prepare microcapsules, it may be desirable to have a homogenization speed of 500 to 3000 rpm and a homogenization time of 5 to 10 minutes.

In the step (2-2), the hydrophobic outer wall precursor may mean a hydrophobic material forming the outer wall of the capsule. The capsule includes an outer wall and an interior surrounded by the outer wall. Therefore, when the capsule is prepared by the above method, the hydrophobic outer wall precursor contained in the second solution forms the outer wall of the capsule, and the oil extract contained in the first solution may be supported inside the capsule. Since the outer wall of the capsule is formed of a hydrophobic material, physical properties required for manufacturing and using the capsule, such as durability and heat resistance, can be satisfied.

In the step (2-2), the hydrophobic outer wall precursor may be a hydrophobic polymer material. The hydrophobic outer wall precursor may be one selected from the group consisting of melamine-formaldehyde, polyurethane, silicone, polyurethane-silicone hybrid, polystyrene, and nylon, but is not limited thereto. The hydrophobic outer wall precursor may be melamine-formaldehyde. The melamine-formaldehyde may mean a mixture of melamine and formaldehyde. The polyurethane-silicone hybrid may mean a hybrid resin-based polymer material of polyurethane and silicone.

In the step (2-2), the second solution may be composed of a hydrophobic outer wall precursor. The second solution may be composed of melamine-formaldehyde. For example, the second solution may be prepared by stirring melamine and formaldehyde. The stirring may be performed at 80 to 90 ° C, but is not limited thereto.

In the step (2-3), a person skilled in the art can select an appropriate method for the crosslinking reaction in consideration of the type of hydrophobic outer wall precursor, the type of surfactant, and the like. For example, the crosslinking reaction may be a melamine-formaldehyde crosslinking reaction, but is not limited thereto.

Capsules prepared by the method may be microcapsules or nanocapsules. A microcapsule may refer to a capsule having a particle diameter in micro units. A nanocapsule may refer to a capsule having a particle size of nanometers. Accordingly, the capsule may have a size of 0.1 to 900 μm, 0.1 to 500 μm, 0.1 to 300 μm, 0.1 to 200 μm, 0.1 to 100 μm, 0.1 to 50 μm, 0.1 to 20 μm, 0.1 to 10 μm, 0.1 to 5 μm, 0.5 to 900 μm, 0.5 to 500 μm, 0.5 to 300 μm, 0.5 to 200 μm, 0.5 to 100 μm, 0.5 to 50 μm, 0.5 to 20 μm, 0.5 to 10 μm, 0.5 to 5 μm, 1 to 900 μm, 1 to 500 μm, 1 to 300 μm, 1 to 200 μm, 1 to 100 μm, 1 to 50 μm, 1 to 20 μm, 1 to 10 μm, or 1 to 5 μm.

The capsule prepared by the above method may have a hydrophobic outer wall. The capsule prepared by the above method can stably encapsulate the oil extract of the strain.

The capsules prepared by the above method may be sustained-release capsules. Thus, the capsule can slowly and continuously release the substance contained therein. The capsule can slowly and continuously release the oil extract of the strain contained therein.

Another aspect provides a capsule loaded with an oil extract of a strain of Streptococcus infantis .

Details of the capsule are as described above.

The capsule may be manufactured by the method according to the above aspect.

The capsules may be microcapsules or nanocapsules. Accordingly, the capsule may have a size of 0.1 to 900 μm, 0.1 to 500 μm, 0.1 to 300 μm, 0.1 to 200 μm, 0.1 to 100 μm, 0.1 to 50 μm, 0.1 to 20 μm, 0.1 to 10 μm, 0.1 to 5 μm, 0.5 to 900 μm, 0.5 to 500 μm, 0.5 to 300 μm, 0.5 to 200 μm, 0.5 to 100 μm, 0.5 to 50 μm, 0.5 to 20 μm, 0.5 to 10 μm, 0.5 to 5 μm, 1 to 900 μm, 1 to 500 μm, 1 to 300 μm, 1 to 200 μm, 1 to 100 μm, 1 to 50 μm, 1 to 20 μm, 1 to 10 μm, or 1 to 5 μm.

The content of the oil extract can be appropriately selected by those skilled in the art within a range capable of exhibiting skin functional efficacy without substantially affecting the formulation stability of the capsule. The oil extract is 0.001 to 90% by weight, 0.001 to 80% by weight, 0.001 to 70% by weight, 0.001 to 60% by weight, 0.001 to 50% by weight, 0.001 to 40% by weight, 0.001 to 30% by weight based on the total weight of the capsule. %, 0.001 to 20% by weight, 0.001 to 10% by weight, or 0.001 to 5% by weight.

The capsule can release the active ingredient contained therein over a long period of time and continuously. For example, the capsule may continuously release the active ingredient contained therein for 2 weeks or more, 4 weeks or more, 8 weeks or more, 12 weeks or more, 16 weeks or more, 20 weeks or more, or 24 weeks or more.

Another aspect provides the fiber to which the capsule according to the above aspect is attached.

The fiber is prepared by (a) preparing a processing solution containing the capsule and a binder resin;

(b) immersing and stirring the fibers in the processing solution; and

(c) curing and drying the resulting product of step (b) to produce fibers with attached capsules.

The total content of the capsule is 0.1 to 20% by weight, 0.1 to 15% by weight, 0.1 to 10% by weight, 0.2 to 20% by weight, 0.2 to 15% by weight, 0.2 to 10% by weight, 0.2 to 10% by weight based on the total weight of the fiber. to 5% by weight, 0.2 to 2% by weight, 0.5 to 20% by weight, 0.5 to 15% by weight, 0.5 to 10% by weight, 0.5 to 5% by weight, or 0.5 to 2% by weight, preferably 2.0 to 2% by weight. 5.0% by weight. When the content of the capsule is lower than the above range, the effect of the supported component may be insignificant, and when the content is higher than the above range, the touch feeling of the product is deteriorated by the attached capsule, which is not preferable.

The content of the binder resin is 1 to 10% by weight, 1 to 8% by weight, 1 to 6% by weight, 1 to 5% by weight, 1 to 4% by weight, 2 to 10% by weight, 2 to 10% by weight based on the total weight of the fiber. to 8% by weight, 2 to 6% by weight, 2 to 5% by weight, or 2 to 4% by weight, preferably 2 to 5% by weight. If the amount is lower than this range, the amount of capsule attachment may be small, and if it is higher than this range, the touch feeling becomes hard and yellowing occurs, which is not preferable.

In the step (a), examples of the binder resin include an acrylic resin, a urethane resin, a dimethylol dihydroxy ethylene urea resin, and a dimethyl urea glyoxal resin. The binder resin may be an acrylic resin.

In the step (a), the processing solution may mean a fiber processing solution. The fiber processing solution may refer to a solution capable of processing fibers.

In the step (b), the fiber is a natural or man-made linear object that is long and thin and softly bendable. The fibers may be natural fibers or man-made fibers. Types of the natural fibers or artificial fibers are known. For example, the fiber may be cotton fiber, but is not limited thereto.

In the step (b), the immersion and stirring may be performed using a conventional method.

In the step (c), the curing and drying may be performed using conventional methods. For example, the curing and drying may be performed at a temperature of 80 to 200 °C, 80 to 180 °C, 100 to 200 °C, 100 to 180 °C, 120 to 200 °C, or 120 to 180 °C. The curing and drying may be performed for 1 to 10 minutes or 1 to 5 minutes.

The fibers may exhibit functionality by attaching capsules. The functionalities may be skin functionalities. The skin functionality may be anti-inflammatory, anti-allergic or inhibition or relief of itching. Accordingly, the fiber to which the capsule is attached may be a functional fiber or a skin functional fiber. The fiber to which the capsule is attached may be a cosmetic fiber. When bedding or clothing is manufactured using the cosmetic fiber, the active ingredient released from the capsule can be delivered to the skin in contact with the bedding or clothing.

Another aspect provides bedding or clothing comprising the fiber according to the one aspect.

The bedding may include pillows, duvets, bed sheets, mattress covers, etc., but is not limited thereto.

The clothing may include underwear, outerwear, etc., but is not limited thereto.

Since the bedding or clothing is manufactured using the fiber to which the capsule is attached, the active ingredient released from the capsule can be delivered and penetrated into the skin in contact with the bedding or clothing. Accordingly, the bedding or clothing may be functional (eg, skin functional) bedding or functional (eg, skin functional) clothing.

Redundant content is omitted in consideration of the complexity of the present specification, and terms not defined otherwise in the present specification have meanings commonly used in the technical field to which the present invention belongs.

According to the method according to one aspect, the skin functional material of the strain of Streptococcus infantis can be supported in the capsule.

In addition, the capsule prepared by the above method can continuously release the oil extract component of the strain of Streptococcus infantis over a long period of time. Therefore, by attaching the capsule to the fiber, it is possible to prepare a skin functional fiber having effects such as anti-inflammatory, anti-atopic, and anti-itching. These skin functional fibers can be applied to bedding or clothing to deliver active ingredients to the skin.

1 is a scanning electron micrograph of the capsule prepared in Example 2.

2 is a scanning electron micrograph of the functional fiber to which the capsules are attached prepared in Example 3.

3 is a scanning electron micrograph of the functional fiber to which the capsules are attached prepared in Example 3.

Figure 4 is a graph showing the absorbance change of the active ingredient (jojoba oil) released from the capsule over time.

5 is a graph showing the change in absorbance of the active ingredient (jojoba oil) released from the capsule over time and a formula for calculating it.

6 is a result showing the relative expression level of IL-1α in keratinocytes treated with CX-4 strain culture medium.

7 is a result showing the relative expression level of TSLP in keratinocytes treated with CX-4 strain culture medium.

Figure 8 is a result of confirming whether or not cytotoxicity by treatment of the surface to which the strain oil extract capsule is attached in keratinocytes. (-): untreated control, Con: cotton-treated group without capsules, jojoba: cotton-treated group with CX-4 strain jojoba oil extract capsules attached, macadamia: CX-4 strain macadamia nut oil extract capsules attached cotton treatment group.

9 is a result showing the relative expression level of IL-1α in keratinocytes treated with cotton to which the CX-4 strain oil extract capsule was attached.

10 is a result showing the relative expression level of TSLP in keratinocytes treated with cotton to which CX-4 strain oil extract capsules are attached.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

Example 1. Preparation of Oil Extracts of Streptococcus infantis strains

An extract of strain Streptococcus infantis CX-4 was prepared using oil as an extraction solvent.

Specifically, Streptococcus infantis CX-4 strain (accession number: KCCM12642P) was inoculated to 1% so that the number of bacteria was 1.0Х10 ⁹ in the medium. After inoculation, it was incubated at 37°C for 120 hours. After completing the culture, it was confirmed that the pH was formed to 4.0 ± 0.2, and when it was confirmed that the color was dark brown with the naked eye, the culture was stopped. Cells were obtained by centrifugation. The obtained cells were added to oil at 1 to 5% (v/v), stirred at 45 to 55° C. for 12 hours, and then extracted at room temperature for 24 hours. Thereafter, bacteria were removed by centrifugation and filtration with a 0.2 μm filter to obtain an oil extract of Streptococcus infantis CX-4 strain.

Jojoba oil was used as the oil, but other types of oil such as green tea seed oil and macadamia nut oil may be used instead of jojoba oil.

Example 2. Preparation of capsules containing strain oil extract

A capsule containing the oil extract of the Streptococcus infantis CX-4 strain of Example 1 was prepared.

Specifically, polyglycol ether as a nonionic surfactant was added to the solvent in an amount of 3% by weight based on the total weight of the first solution (emulsion), followed by stirring. The oil extract of the CX-4 strain of Example 1 was added thereto and homogenized at 1000 to 3000 rpm for 5 to 10 minutes to prepare a first solution. In addition, a second solution was prepared by stirring melamine and formaldehyde at 80 to 90 ° C.

A 0.1M NaOH aqueous solution was added to the mixture of the first solution and the second solution, the pH was adjusted to 9-10, and the mixture was stirred at 50° C. for 1 hour. Thereafter, the temperature was raised to 70 ° C., and the pH was adjusted to 5-6 by adding 10M acetic acid and reacted for 5 hours to prepare microcapsules through a crosslink reaction of melamine-formaldehyde.

1 is a scanning electron micrograph of the capsule prepared in Example 2. As shown in FIG. 1, the particle size of the prepared capsules was confirmed to be about 1 to 5 μm. Thus, it was confirmed that the microcapsules were prepared.

On the other hand, when preparing the first solution, (i) use sodium dodecyl sulfate surfactant instead of polyglycol ether, (ii) increase the content of surfactant to 5 to 7% by weight, (iii) homogeneous RPM If it is increased to 5000-10000 rpm, it is possible to manufacture nanocapsules.

Example 3. Preparation of functional fibers with attached capsules

Functional fibers to which the capsules of Example 2 were attached were prepared.

Specifically, a processing solution was prepared by adding the capsule of Example 2 to an acrylic binder resin. Bedding fibers such as bedding cotton or covers were immersed and stirred in the processing solution. Cured and dried at 120 to 180 ° C. for 1 to 5 minutes to prepare functional fibers with attached capsules. In this embodiment, cotton fiber was used as an exemplary type of fiber. Bedding such as pillows and duvets can be manufactured using the manufactured fibers.

The total content of the capsule is 0.5 to 10% by weight based on the total weight of fibers. The content of the binder resin is 1 to 5% by weight based on the total weight of the fiber.

2 is a scanning electron micrograph of the functional fiber to which the capsules are attached prepared in Example 3.

3 is a scanning electron micrograph of the functional fiber to which the capsules are attached prepared in Example 3.

As shown in Figures 2 and 3, it was confirmed that the capsules were well attached to cotton fibers commonly used in bedding or clothing.

Experimental Example 1. Confirmation of sustained release performance of capsules

An experiment was conducted to confirm the sustained release performance of the capsule prepared in Example 2.

Specifically, 0.5 g of the capsule prepared in Example 2 was weighed and placed in 100 g of ethanol as a solvent. While stirring, the active substance eluted from the capsule was collected over time, filtered through a syringe filter, and absorbance was measured using a UV-Vis Spectrophotometer. The release behavior of oil, an active substance, from the capsule was confirmed through the change in absorbance.

Figure 4 is a graph showing the change in absorbance of the active ingredient (jojoba oil) released from the capsule over time.

5 is a graph showing the change in absorbance of the active ingredient (jojoba oil) released from the capsule over time and a formula for calculating it.

As shown in FIGS. 4 and 5, through the confirmation of absorbance change of the capsule prepared in Example 2, it was confirmed that the size of the peak increased over time in the same wavelength band as the oil contained in the capsule. In addition, as a result of showing this as a trend line, it was found that the active ingredient inside the capsule was continuously released over time.

Therefore, when bedding or clothing is manufactured using the functional fiber to which the capsule of Example 2 is attached, the strain oil extract contained in the capsule is released for a long period of time and continuously when sleeping or wearing clothing, thereby continuously providing efficacy to the skin. I knew that it could work.

Experimental Example 2. Analysis of anti-inflammatory, anti-atopic and itching inhibitory activity of the strain culture medium

Whether or not the CX-4 strain culture medium had anti-inflammatory, anti-atopic and anti-itching activities was analyzed.

Specifically, HaCaT cells, a human keratinocyte, were cultured in DMEM medium (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) containing 10% fetal bovin serum. % CO ₂ was performed in an incubator. The medium was exchanged every 3 to 4 days, and subculture was performed when the cells proliferated excessively. Thereafter, the cells were dispensed at 5×10 ⁵ /well and washed with phosphate buffered saline solution (PBS) after 24 hours of culture. Cells were seeded into each well containing DMEM medium without FBS, and inflammation was induced in the keratinocyte cell line by adding 10 μg/ml of Poly I:C and 10 ng/ml of IL-4. Next, the culture solution of the CX-4 strain was treated at a concentration of 0.1% (w/w) or 1% (w/w), and then further cultured for 4 hours. As a positive control, 1 μM of dexamethasone was used. As a negative control, a medium to which the CX-4 strain was not added was used. The expression levels of IL-1α and TSLP were analyzed by RT-PCR using primers for IL-1α and TSLP (thymic stromal pymphopoietin). The expression level of the gene was finally analyzed through correction for the β-actin gene, and the results are shown in FIGS. 6 and 7, respectively.

6 is a result showing the relative expression level of IL-1α in keratinocytes treated with CX-4 strain culture medium.

7 is a result showing the relative expression level of TSLP in keratinocytes treated with CX-4 strain culture medium.

As shown in Figure 6, it was confirmed that the CX-4 strain culture significantly reduced the expression of the inflammatory cytokine IL-1α compared to the untreated control group.

In addition, as shown in Figure 7, it was found that the CX-4 strain culture medium reduced the expression of the atopy and itch inhibitor TSLP compared to the untreated control group.

Experimental Example 3. Confirmation of Cytotoxicity of Strain Oil Extract

Whether or not the CX-4 strain oil extract was cytotoxic was analyzed.

Specifically, the surface to which the CX-4 strain oil extract capsules prepared in the same manner as in Example 3 was attached was prepared. At this time, the oil used in preparing the CX-4 strain oil extract was jojoba oil or macadamia oil.

Next, the surface to which the capsule was attached was treated with 5 sheets of human keratinocyte HaCaT cells in a size of 2 cm 2 . Apoptotic cells were confirmed after 24 hours of cotton treatment, and the results are shown in FIG. 8 .

Figure 8 is a result of confirming whether or not cytotoxicity by treatment of the surface to which the strain oil extract capsule is attached in keratinocytes.

As shown in FIG. 8, it was confirmed that the CX-4 strain oil extract, the capsule carrying the extract, and the side to which the capsule was attached did not have cytotoxicity.

Therefore, it was found that the functional fibers to which the CX-4 strain oil extract capsules are attached can be safely used.

Experimental Example 4. Anti-inflammatory, anti-atopic, anti-itch activity analysis of strain oil extract

It was analyzed whether the side to which the CX-4 strain oil extract capsules were attached had anti-inflammatory, anti-atopic and anti-itching activities. Types of oil were typically jojoba oil and macadamia nut oil.

Specifically, IL- The expression levels of 1α and TSLP were analyzed.

9 is a result showing the relative expression level of IL-1α in keratinocytes treated with cotton to which the CX-4 strain oil extract capsule was attached.

10 is a result showing the relative expression level of TSLP in keratinocytes treated with cotton to which CX-4 strain oil extract capsules are attached.

As shown in Figure 9, it was confirmed that the cotton-treated group to which the CX-4 strain oil extract capsules were attached significantly reduced the expression of the inflammatory cytokine IL-1α compared to the untreated control group.

In addition, as shown in FIG. 10, it was found that the cotton-treated group to which the CX-4 strain oil extract capsules were attached reduced the expression of the atopy and pruritus inhibitory factor TSLP compared to the untreated control group.

These results suggest that the CX-4 strain, the CX-4 strain oil extract, the capsule containing the CX-4 strain oil extract, or the functional fiber to which the capsule is attached can be usefully used to prevent, improve, or treat skin inflammatory diseases. In addition, it means that it can be usefully used for the prevention, improvement, or treatment of atopy, or the prevention, treatment, or improvement of itching (pruritus).

Taken together, the capsule loaded with the oil extract of strain Streptococcus infantis CX-4 can release the extract components loaded therein over a long period of time and continuously. In addition, the oil extract of strain CX-4 has anti-inflammatory, anti-atopic and anti-itching activities. Therefore, when bedding or clothing is manufactured using the functional fiber to which the capsule is attached, the strain oil extract contained in the capsule is released for a long time and continuously to continuously improve the skin such as anti-inflammatory, anti-atopic and itching inhibition. can be effective.

The above description is merely an example of the technical idea of the present invention, and various modifications and variations can be made to those skilled in the art without departing from the essential characteristics of the present invention.

<Access number>

Name of depository organization: Korea Center for Microbial Conservation (overseas)

Accession number: KCCM12642P

Entrusted date: 20191218

Claims (11)

1. (1) using oil as an extraction solvent to prepare an oil extract of Streptococcus infantis strain; and

   (2) comprising the step of preparing a capsule containing the oil extract,

   Streptococcus infantis ( Streptococcus infantis ) A method for producing a capsule containing an oil extract of the strain.
2. The method of claim 1, wherein the oil is a vegetable oil.
3. The method according to claim 1, wherein the strain is Streptococcus infantis deposited under accession number KCCM12642P ( Streptococcus infantis ) CX-4 strain method.
4. The method according to claim 1, wherein the strain comprises 16S rRNA having 98% or more sequence identity with SEQ ID NO: 1.
5. The method according to claim 1, wherein the step (2),

   (2-1) The oil extract is added to a solution containing one surfactant selected from the group consisting of sodium dodecyl sulfate, sodium dodecylbenzenesulfonate, polyglycol ether and stearyl methacrylate and homogenized to prepare 1 preparing a solution;

   (2-2) preparing a second solution containing one hydrophobic outer wall precursor selected from the group consisting of melamine-formaldehyde, polyurethane, silicone, polyurethane-silicone hybrid, polystyrene and nylon; and

   (2-3) preparing a capsule by mixing and cross-linking the first solution and the second solution.
6. Streptococcus infantis ( Streptococcus infantis ) Capsule containing oil extract of the strain.
7. The capsule according to claim 6, wherein the capsule is prepared by the method according to any one of claims 1 to 5.
8. The method according to claim 6, wherein the capsule has a particle diameter of 0.1 to 10 μm.
9. A fiber to which the capsule according to claim 6 is attached.
10. Bedding comprising the fiber according to claim 9.
11. A garment comprising the fiber according to claim 9 .

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