Use of kestose for improving skin lipids or skin microflora

Abstract

The present invention relates to a composition comprising kestose as an active ingredient, and a use of the composition for improving skin barrier lipids or skin microflora or for preventing, treating, or alleviating skin lipid- or skin microflora-related diseases.

Description

Use of Kestos to improve skin lipids or skin microbiota

[0001] The present invention relates to a composition comprising kestos as an active ingredient, and to the use of the composition for improving skin barrier lipids or improving the overall skin microbiota, or for preventing, treating, or improving diseases related to skin lipids or skin microbiota.

[0002] The skin is typically characterized by three distinct layers: the stratum corneum, the epidermis, and the dermis. The stratum corneum, the outermost layer, is composed of keratinized cells surrounded by lipid-filled intercellular spaces. The stratum corneum provides a substantial physical barrier to the penetration of most substances into the lower layers of the skin. However, in addition to preventing the transport of substances to other skin layers, this layer also helps prevent water loss from the skin. Both functions are primarily due to the presence of lipids within the stratum corneum. The composition and organization of lipids in the stratum corneum are essential for its barrier function.

[0003] The intercellular lipid domain of the stratum corneum is primarily composed of cholesterol (CHOL), free fatty acids (FFAs), and ceramides (CER) in equal proportions. CER is a structurally heterogeneous group of sphingolipids, consisting of a sphingoid base linked to a fatty acid. Ceramides exist in various subclasses depending on the structure of the fatty acid that binds to the precursor sphingoid. Currently, 15 types of ceramides have been discovered in the skin. EOS ceramide (CER[EOS]) refers to a ceramide in which an ester-linked omega-hydroxy group (EO) is attached to a sphingosine base as a fatty acid.

[0004] Accordingly, in order to improve skin lipid and skin microbiome, the components that make up skin lipid and their composition are important, and in particular, it is necessary to increase the proportion of EOS ceramides (CER[EOS]) that have relatively long-chain fatty acids and to develop materials that increase beneficial microorganisms in the skin microbiome.

[0005] One example of the present invention relates to a composition for controlling the content ratio of EOS ceramides in skin, for example, the stratum corneum of the skin, including kestos, or a method for controlling the content ratio of EOS ceramides.

[0006] One example of the present invention relates to a composition for increasing the content or ratio of EOS ceramides having long-chain fatty acids in the skin, comprising kestose, and a method for increasing the content ratio of EOS ceramides having long-chain fatty acids in the skin, comprising fatty acids having C31 to C36, using kestose.

[0007] One example of the present invention relates to a composition for reducing the content or ratio of EOS ceramides having short-chain fatty acids in the skin, including fatty acids having C28 to C30, comprising kestose, and a method for reducing the content ratio of EOS ceramides having short-chain fatty acids in the skin using kestose.

[0008] One example of the present invention relates to a composition for increasing beneficial microorganisms, for example, microorganisms of the genus Prevotella, on the skin, comprising kestos, and a method for increasing the content or ratio of beneficial microorganisms, for example, microorganisms of the genus Prevotella, on the skin using kestos.

[0009] Another embodiment of the present invention relates to a composition comprising kestos for reducing, improving, or preventing sleep disturbances caused by skin problems. Another embodiment of the present invention relates to a composition comprising kestos for improving skin conditions.

[0010] The present invention relates to a composition comprising kestos as an active ingredient, the use of the composition for improving skin barrier lipids or improving skin microbiota, or for preventing, treating or improving diseases related to skin lipids or skin microbiota, and for improving skin conditions.

[0011] Furthermore, the present invention relates to a method for preventing or treating diseases related to skin lipids or skin microbiota, comprising kestose as an active ingredient. Furthermore, the present invention relates to a method for improving skin condition, comprising kestose as an active ingredient.

[0012] One example of the present invention relates to a composition for controlling the content ratio of EOS ceramides in skin, for example, the stratum corneum of the skin, including kestos, or a method for controlling the content ratio of EOS ceramides.

[0013] Specifically, the present invention relates to a composition for increasing the content ratio of EOS ceramides having long-chain fatty acids in the skin, for example, in the stratum corneum, including C31 to C36 fatty acids, and a method for increasing the content ratio of EOS ceramides having long-chain fatty acids in the skin, for example, in the stratum corneum, among total EOS ceramides, using kestose, wherein, for example, the EOS ceramides having long-chain fatty acids in the skin, for example, in the stratum corneum, including C31 to C36 fatty acids, may be at least one selected from the group consisting of 31:0-EOS ceramide and 32:0-EOS ceramide.

[0014] One example of the present invention relates to a composition for reducing the content ratio of EOS ceramides having short-chain fatty acids in skin, for example, skin containing fatty acids having C28 to C30 in the stratum corneum, including kestose, and a method for reducing the content ratio of EOS ceramides having short-chain fatty acids in skin, for example, skin containing fatty acids having C28 to C30 in the stratum corneum, using kestose, wherein, for example, EOS ceramides having fatty acids in the stratum corneum may be at least one selected from the group consisting of 28:0-EOS ceramide and 30:0-EOS ceramide.

[0015] The above total EOS ceramide may include, for example, total EOS ceramides contained in skin lipids, for example, at least one EOS ceramide selected from the group consisting of d18:1-c28:0-18:2, d18:1-c29:0-18:2, d18:1-c30:0-18:2, d18:1-c31:0-18:2, d18:1-c32:0-18:2, d18:1-c33:0-18:2, d18:1-c34:0-18:2, d18:1-c35:0-18:2, and d18:1-c36:0-18:2. Specifically, the entire EOS ceramide may be an EOS ceramide including the d18:1-c28:0-18:2, d18:1-c29:0-18:2, d18:1-c30:0-18:2, d18:1-c31:0-18:2, d18:1-c32:0-18:2, d18:1-c33:0-18:2, d18:1-c34:0-18:2, d18:1-c35:0-18:2, and d18:1-c36:0-18:2.

[0016] One example of the present invention relates to a composition for increasing beneficial microorganisms, for example, microorganisms of the genus Prevotella, on the skin, comprising kestos, and a method for increasing the content or ratio of beneficial microorganisms, for example, microorganisms of the genus Prevotella, on the skin using kestos.

[0017] A further example of the present invention relates to a composition for reducing, improving or preventing sleep disturbance due to skin trouble, comprising kestos.

[0018] A further example of the present invention relates to a method for improving sleep disturbance due to skin trouble, comprising ketose.

[0019]

[0020] Hereinafter, the present invention will be described in more detail.

[0021] The present invention relates to a composition comprising kestose for controlling the content ratio of EOS ceramides in the skin, for example, in the stratum corneum of the skin, or to a method for controlling the content ratio of EOS ceramides, and more particularly to the use of kestose for increasing the content or ratio of EOS ceramides having long-chain fatty acids in the skin, comprising fatty acids having C31 to C36, and / or for decreasing the content or ratio of EOS ceramides having short-chain fatty acids in the skin, comprising fatty acids having C28 to C30.

[0022] The above-mentioned skin may be at least one selected from the group consisting of EOS ceramides having long-chain fatty acids, specifically 31:0-EOS ceramide and 32:0-EOS ceramide. In addition, the above-mentioned skin may be at least one selected from the group consisting of EOS ceramides having short-chain fatty acids, specifically 28:0-EOS ceramide and 30:0-EOS ceramide.

[0023] Specifically, as a result of skin lipid analysis of skin samples collected before and after consuming Kestos according to the present invention, a decrease in 28:0-EOS ceramide (EOS d18:1 / 28:0 / 18:2) and 30:0-EOS ceramide (EOS d18:1 / 30:0 / 18:2) and an increase in 32:0-EOS ceramide (EOS d18:1 / 32:0 / 18:2) were confirmed in the test group.

[0024] In addition, in the present invention, the content of relatively long-chain 31:0 ceramide (EOS d18:1 / 31:0 / 18:2) and 32:0 ceramide (EOS d18:1 / 32:0 / 18:2) increased. In addition, in the present invention, the content of relatively short-chain 28:0 or 30:0 ceramide decreased, and it was confirmed that ceramide has an effect on skin diseases or skin troubles because skin itching and sleep disturbance were reduced at that time. Therefore, in the present invention, after ingestion of kestose, 32:0-EOS ceramide (EOS d18:1 / 32:0 / 18:2), which is a relatively long-chain fatty acid, increases, and 28:0-EOS ceramide (EOS d18:1 / 28:0 / 18:2) and 30:0-EOS ceramide, which are relatively short-chain fatty acids, decrease, thereby reducing moisture loss from the skin, thereby exhibiting beneficial effects.

[0025] Ceramides are representative moisture-retaining substances that surround the outer surface of keratinocytes. In atopic dermatitis, ceramides in the stratum corneum are reduced. Reduced synthesis of ceramides, a key component of skin barrier lipids, causes disruption of lamellae formation and lipid lamellar structure formation, as well as a decrease in overall epidermal lipids, leading to abnormalities in skin barrier function. EOS ceramides play a crucial role in the pathophysiology of atopic dermatitis due to their high hydrophobicity, and the length of the fatty acid chain is important for maintaining skin barrier function. Short fatty acid chains in atopic dermatitis skin increase skin barrier permeability, disrupting skin barrier function and worsening skin symptoms.

[0026] In particular, compounds in which ester-linked omega-hydroxyacyl chains are attached to sphingosine bases, called ceramide EOS (CER EOS) or EOS ceramides, are essential for the lipid structure of the skin barrier. In particular, omega-O-acyl ceramides, such as 32-linoleoyloxydotriacontanoyl sphingosine (CER[EOS]), are essential components of the lipid skin barrier, which protects our bodies from excessive water loss and the penetration of unwanted substances. These ceramides induce lipid assembly into the epidermal specific long-period phase (LPP).

[0027] Specifically, EOS ceramide has a structure (-NH-CO-) in which a carboxyl group (-COOH) of a fatty acid is bonded to an amino group (-NH2) of a sphingosine base. Specifically, when the fatty acid portion of the ceramide is an ω-hydroxy fatty acid and an additional fatty acid is ester-bonded to the hydroxyl terminal, it is particularly called an acylceramide (ω-O-acylceramide), i.e., EOS ceramide. The ω-hydroxy fatty acid may have 28 to 36 carbon atoms and may be a fatty acid having 0 or 1 double bond between carbons, with a fatty acid having 0 (i.e., a saturated fatty acid) being more preferable. Examples of the fatty acid bonded to the ω-hydroxy terminal include stearic acid, oleic acid, linoleic acid, and linolenic acid. The ceramide of the present invention preferably contains an acylceramide in which the fatty acid bonded to the ω-hydroxy terminal is linoleic acid. The sphingosine is a type of sphingoid base, has two hydroxyl groups, 18 carbon atoms, and 1 double bond between carbon atoms.

[0028] An example of the EOS ceramide according to the present invention may be at least one selected from the group consisting of d18:1-c28:0-18:2, d18:1-c29:0-18:2, d18:1-c30:0-18:2, d18:1-C31:0-18:2, d18:1-c32:0-18:2, d18:1-c33:0-18:2, d18:1-c34:0-18:2, d18:1-c35:0-18:2, and d18:1-c36:0-18:2.

[0029] Another example of the present invention relates to a composition for increasing beneficial microorganisms, for example, microorganisms of the genus Prevotella, on the skin, comprising kestos, and a method for increasing the content or ratio of beneficial microorganisms, for example, microorganisms of the genus Prevotella, on the skin using kestos.

[0030] Specifically, the present invention performed a skin microbiome profile analysis and species diversity analysis using skin samples obtained before and after ingestion of Kestos to confirm the species diversity and profiling of the skin microbiome, and as a result, it was confirmed that the alpha diversity Chao1 diversity index significantly increased after ingestion of Kestos, and skin microbiome profiling was confirmed through taxonomic composition at the genus level. At the genus level, it was confirmed that microorganisms in the genus Prevotella significantly increased after ingestion of Kestos.

[0031] The composition according to the present invention can improve skin condition and sleep disorders caused by skin diseases by controlling the skin EOS ceramide ratio / content, increasing the species diversity of the skin microbiome, and increasing the number of microorganisms of the genus Prevotella. The improvement in skin condition can be, for example, improvement in skin itching, improvement in atopy, enhancement of skin immune function, reduction in inflammatory cytokines, improvement in immune imbalances such as Th1, Th2, and Th17, reduction in skin moisture loss, reduction in skin permeability, enhancement of the skin barrier, improvement in skin dryness, and dermatitis.

[0032] The composition according to the present invention comprises kestose or 1-kestose as an active ingredient, and kestose can be used alone or in a sugar composition containing it. Kestose can be used in liquid or powder form, and the powder can be amorphous or crystalline. The kestose can be purchased and used as a commercial product, or can be manufactured using a predetermined raw material.

[0033] An example of the above sugar composition may be a kestose-containing fructooligosaccharide (FOS). FOS is composed of a linear chain having 1 to 9 fructose residues linked to a sucrose molecule by a β2→1 bond. The kestose-containing fructooligosaccharide preferably has 1-kestose as the main component, and may, for example, contain a high content of 1-kesotse (GF2), and may contain at least one selected from the group consisting of nystose (GF3) and 1-F-fructosyl nystose (GF4).

[0034] There is no particular limitation on the kestose or sugar composition containing kestose that can be used in the present invention, and it can be produced using sugar as a substrate, usually by using an enzyme having kestose conversion activity or a microorganism that produces the enzyme.

[0035] The enzyme having the above kestose conversion activity is an enzyme having an activity of converting a fructooligosaccharide containing kestose from a sugar-containing substrate, and may be an enzyme derived from at least one selected from the group consisting of, for example, an Aspergillus niger strain, a Pichia farinose strain, a Yarrowia lipolytica strain, a Millerozyma farinose strain, and an Aspergillus oryzae strain.

[0036] In the present invention, the intake (administration) of kestose is, for example, 0.01 to 0.34 g / kg body weight, preferably 0.01 to 0.30 g / kg body weight, more preferably 0.01 to 0.24 g / kg body weight, 0.03 to 0.34 g / kg body weight, 0.05 to 0.34 g / kg body weight, 0.06 to 0.34 g / kg body weight, 0.03 to 0.24 g / kg body weight, 0.05 to 0.24 g / kg body weight, 0.06 to 0.24 g / kg body weight, 0.03 to 0.30 g / kg body weight, 0.05 to 0.30 g / kg body weight, 0.06 to Examples include 0.30 g / kg body weight, 0.03 to 0.26 g / kg body weight, 0.05 to 0.26 g / kg body weight, or 0.06 to 0.26 g / kg body weight. This intake amount is not limited to once a day, but can be divided into multiple doses and taken.

[0037] One method is to administer ketose orally to humans or animals, either as is or in the form of food or medicine.

[0038] The above Kestos composition may be a composition containing a total of 1 g to 30 g, 1 g to 20 g, 1 g to 15 g, 2 g to 15 g, 1 g to 10 g, 2 g to 10 g, 1 g to 7 g, or 2 g to 7 g of Kestos as an active ingredient per day for a 60 kg adult.

[0039] In addition, the above-mentioned kestose composition may be a composition containing a combined content of kestose, nystose, and fructofuranosylnystose of 400 mg / g or more, 500 mg / g or more, 800 mg / g or more, or 900 mg / g or more.

[0040] The kestose usable in the present invention may be in either liquid or powder form, and may be included in the composition according to the present invention in various amounts. The kestose may be used as a single component or in a mixed composition containing other sugars, such as fructooligosaccharide (FOS).

[0041] The above-mentioned liquid or powdered kestos may be a composition having a content of 50(w / w) % or more, 55(w / w) % or more, 60(w / w) % or more, 65(w / w) % or more, 70(w / w) % or more, 75(w / w) % or more, preferably 80(w / w) % or more, 85(w / w) % or more, or 87(w / w) % or more, based on the total solid content. In addition, the crystalline kestos may be a composition having a content of 98% or more based on the total solid content.

[0042] The dosage forms of pharmaceuticals, quasi-drugs, and supplements containing Kestos are not particularly limited, and a dosage form appropriate for the administration method can be appropriately selected. For example, in the case of oral administration, solid or liquid dosage forms such as powders, tablets, sugar syrups, capsules, granules, dry syrups, liquids, syrups, drops, and drinks can be used.

[0043] Kestose can be added to and used in the normal manufacturing processes of various foods, food additives, and animal feed. 1-Kestose has a sweetness index of 30, and its taste, physical properties, and processability are close to sucrose. Therefore, it can be used in various foods, beverages, food additives, medicines, or feeds by treating it in the same way as sugar, such as by replacing some or all of the sugar with 1-Kestose in the manufacturing process of various foods.

[0044] Specific embodiments of the composition according to the present invention include, for example, beverages, dairy products, granules for food use, pastes, seasonings, retort foods, baby foods, fermented foods, preserved foods, processed foods such as processed fishery products, processed meat products, and processed grain products, food additives, health foods, animal feed, etc.

[0045] The present invention relates to a composition comprising kestos as an active ingredient, and to the use of the composition for improving skin barrier lipids or improving the overall skin microbiome, or for preventing, treating, or improving diseases related to skin lipids or skin microbiome, wherein the composition can improve skin conditions or improve sleep disorders caused by skin diseases by controlling the ratio / content of EOS ceramides and increasing the species diversity of skin microbiome and increasing the number of microorganisms of the genus Prevotella.

[0046] Figure 1 is a graph analyzing the fatty acid ratio of skin lipid EOS ceramide according to Kestos intake.

[0047] Figure 2 is a graph analyzing the skin microbiota alpha diversity Chao1 index according to Kestos intake.

[0048] Figure 3 shows the results of analysis of Prevotella genus microorganisms among skin microorganisms according to Kestos intake.

[0049] Figure 4 shows the results of evaluating sleep disturbance scores according to Kestos intake.

[0050] Figure 5 shows the results of analysis of peripheral blood eosinophil counts according to kestos intake.

[0051] Figures 6 to 8 show the results of evaluating the relationship between 28:0-EOS ceramide ((EOS d18:1 / 28:0 / 18:2)) and IGA (Investigator's Global Assessment), the relationship between 30:0-EOS ceramide (EOS d18:1 / 30:0 / 18:2) and skin itching score, and the relationship between the ratio of 30:0-EOS ceramide ((EOS d18:1 / 30:0 / 18:2)) and 32:0-EOS ceramide (EOS d18:1 / 32:0 / 18:2) and skin itching score.

[0052] The present invention will be described in more detail with reference to the following examples, but the scope of the present invention is not intended to be limited to the following examples.

[0053]

[0054] Example 1: Experiment using the Kestos diet

[0055] (1) Test design and sample preparation

[0056] The trial was designed as a 12-week, single-center, double-blind, randomized, controlled food study.

[0057] The test food, packaged in a 3.6g stick pouch, was consumed twice a day, one pouch per time, for 12 weeks. The test group consumed 5g / day (equivalent to 4.25g / day as kestose) of Advanced FOS (Samyang Corporation, fructooligosaccharide-K), and the control group consumed 5g / day of maltose for 12 weeks. All test foods were provided by Samyang Corporation, and each test food was assigned a unique random number, so that the information about the test food was not disclosed to the test participants and researchers. The test substance composition information of the test group and the control group is as follows.

[0058] Both the test and control samples were prepared as white powders, weighing 3.6 g / package, and stored at room temperature (around 25°C). One pack was taken twice a day, 5 g / day as Advanced FOS, and 4.25 g / day as Kestose. The Advanced FOS had a Kestose purity of 87.9 wt%. The maltose powder used as the control contained maltose with a purity of 87 wt%.

[0059] Table 1 below shows the weight percent of each ingredient contained in 3.6 g / package. In the ingredients listed in Table 1 below, anhydrous crystalline glucose was used as an excipient, magnesium stearate was used as a lubricant, and silicon dioxide was used as a flowability improver.

[0060] Ingredient Test Sample (Wt%) Control Sample (Wt%) Main Ingredient Kestose Powder (Kestose Purity 87.9 wt%) 70.0 Maltose Powder (Maltose Purity 87 wt%) 70.0 Anhydrous Crystalline Glucose (Excipient) 26.5 26.5 Magnesium Stearate 2.0 2.0 Silicon Dioxide 1.5 1.5

[0061] (2) Human application test method

[0062] According to the diagnostic criteria of Hanifin and Rajka, the SCORAD score was 15 to 50 (moderate-severe) and the children and adolescents were aged 24 months to 18 years, and voluntarily signed the consent form for the human application study. The subjects were selected through a demographic survey, medical history / medication history survey, physical examination, body measurements (height, weight), vital signs (pulse, body temperature), blood and urine tests, and atopic dermatitis severity assessment. If they met the exclusion criteria, they were enrolled as subjects for the human application study through random assignment. The random assignment, double-blind, control food-controlled human application study, and the allocation ratio of each group was 1:1 for the test group:control group.

[0063] People who met one or more of the following conditions were excluded from the study.

[0064] 1) Those with severe skin disease or systemic disease other than atopic dermatitis

[0065] 2) Those who received systemic antibiotics or systemic corticosteroids within 4 weeks of the start of the human application test

[0066] 3) Those who have been administered systemic immunosuppressants or biological agents within 4 weeks of the start of human application testing

[0067] 4) Those who took oral medications, herbal medicines, health functional foods, probiotics, or prebiotics to improve atopic dermatitis within 1 week of the start of the human application test.

[0068] 5) Those who were administered systemic antihistamines within 2 weeks of the start of the human application test.

[0069] 6) Those who consumed fermented milk or intestinal preparations more than 4 times a week within 1 week of starting the human application test

[0070] 7) Those who received phototherapy within 4 weeks of the start of the human application test

[0071] 8) Those who plan to participate in other human application trials during the human application trial (excluding observational studies)

[0072] 9) Those who participated in another human application test within 3 months of the start of the human application test (excluding observational studies)

[0073] 10) Those with a history of allergy to prebiotics

[0074] 11) Patients deemed unsuitable for human application testing by the investigator

[0075]

[0076] Subjects of the human application study consumed either the test food or the control food for a total of 12 weeks. Screening was conducted at the first visit (Visit 1) to confirm the inclusion / exclusion criteria. At the second visit (Visit 2), subjects were randomly assigned to each group using a block randomization method based on their assigned codes and then distributed the test food. Six weeks (Visit 3) and 12 weeks (Visit 4) after initiating the test food intake, subjects visited the hospital to assess functionality and safety. The trial was reviewed and approved by the Institutional Review Board.

[0077] Forty-eight subjects who met the selection / exclusion criteria for human application trials were randomly assigned to a test group:control group = 1:1 (test group = 24 subjects, control group = 24 subjects) and completed the trial. The human application trial period was planned to be 24 months from the date of approval for human application trials, and the first human application trial subject was screened on May 18, 2021, and the first human application trial subject was enrolled on May 18, 2021. The trial for the last human application trial subject ended on November 22, 2022, and the trial was conducted for approximately 19 months.

[0078]

[0079] (3) Evaluation of human application tests

[0080] Adverse reactions, vital signs (pulse, body temperature), body weight, and clinical pathology tests (hematological / blood chemistry tests, urinalysis) were performed as safety evaluation variables.

[0081] Functional evaluation variables included skin microbiota, skin barrier lipid test, sleep disturbance score, and peripheral blood eosinophil count, and detailed experimental methods are described in Examples 2 to 4.

[0082]

[0083] Example 2: Skin Barrier Lipid Analysis

[0084] (1) Skin barrier lipid analysis method

[0085] Skin lipid samples were obtained using the skin tape stripping method. Specifically, D-squame Skin Sampling Discs (CuDerm, Dallas, TX, USA) with a diameter of 22 mm were used to collect samples at Visits 2 and 4 in Example 1. After repeating the tape disc attachment and removal 20 times each from the lesioned and non-lesioned areas (right anterior cingulate), each disc was attached to D-Squame Standard Storage Cards (CuDerm, Dallas, TX, USA) and stored at -80°C.

[0086] Skin barrier lipids were analyzed from the collected tapes. Specifically, lipids were extracted from the tape discs using 1 mL of MeOH, and analyzed using a Shimadzu LC MS-8060 (Triple Quadrupole Mass Spectrometer). Lipids were separated using a Kinetex C18 column (100 × 2.1 mm, 2.6 μm; Phenomenex, Torrance, CA, USA). Mobile phase A contained 10 mM ammonium acetate in water and methanol (1:9, v / v), and mobile phase B contained 10 mM ammonium acetate in methanol and isopropanol (1:1, v / v). The flow rate was maintained at 0.2 mL / min.

[0087]

[0088] (2) Skin barrier lipid analysis results

[0089] The results of skin lipid analysis of skin samples collected before and after ingestion of the above Kestos are shown in Fig. 1 and the table below, and Fig. 1 is a graph analyzing the ratio of EOS ceramide having a specific fatty acid chain to EOS ceramide having various fatty acid chains contained in skin lipid according to ingestion of Kestos. In the graph of Fig. 1, the horizontal axis represents the type of ceramide to be analyzed as a component of skin lipid, and the horizontal axis represents the content ratio of a specific EOS ceramide based on 100 wt% of the total content of EOS ceramides described on the horizontal axis, expressed as weight percent (wt / wt%).

[0090] EOS Ceramide IngredientsControl Group (Before Intake)Control Group (After Intake)Control Group (After Intake-Before Intake)DifferenceExperimental Group (Before Intake)Experimental Group (After Intake)Experimental Group (After Intake-Before Intake)Difference28:0 EOS-CER(EOS d18:1 / 28:0 / 18:2)4.414.34-0.0714.814.47-0.33629:0 EOS-CER (EOS d18:1 / 29:0 / 18:2)6.236.10-0.1296.566.19-0.37730:0 EOS-CER (EOS d18:1 / 30:0 / 18:2)57.7356.97-0.75858.4756.55-1.91631:0 EOS-CER (EOS d18:1 / 31:0 / 18:2)11.0611.460.40010.5011.040.54032:0 EOS-CER (EOS d18:1 / 32:0 / 18:2)18.6719.320.65117.8919.861.96733:0 EOS-CER (EOS d18:1 / 33:0 / 18:2)0.980.91-0.0710.870.960.08834:0 EOS-CER EOS d18:1 / 34:0 / 18:2)0.900.86-0.0420.880.900.01835:0 E EOS-CER (EOS d18:1 / 35:0 / 18:2)0.010.020.0100.010.020.00436:0 EOS-CER (EOS d18:1 / 36:0 / 18:2)0.000.020.0110.010.020.012Total (weight %)100100-100.00100.00-

[0091] Analysis of skin lipids from skin samples collected before and after taking Kestos showed that the test group showed a significant decrease in the composition ratio of (EOS d18:1 / 28:0 / 18:2) (p=0.029) and (EOS d18:1 / 30:0 / 18:2) (p=0.013), and a significant increase in the composition ratio of (EOS d18:1 / 32:0 / 18:2) (p=0.010). In the control group, the composition ratios of (EOS d18:1 / 28:0 / 18:2) and (EOS d18:1 / 30:0 / 18:2) decreased, and the composition ratio of 32:0-EOS ceramide increased, but the change was not significant. Additionally, the relatively long-chain (EOS d18:1 / 33:0 / 18:2) and (EOS d18:1 / 34:0 / 18:2) contents decreased in the control group and increased in the test group.

[0092] Ceramide is a representative moisture-retaining substance that surrounds the outside of skin keratinocytes, and in atopic dermatitis, ceramide in the stratum corneum is reduced. In particular, the lipid components that account for more than 3% of the total lipid content are five types: (EOS d18:1 / 28:0 / 18:2), (EOS d18:1 / 29:0 / 18:2), (EOS d18:1 / 30:0 / 18:2), (EOS d18:1 / 31:0 / 18:2), and (EOS d18:1 / 32:0 / 18:2). When analyzing the increase and decrease in the content of the five types of EOS ceramides in the experimental and control groups, the relatively short-chain E(18:2)O(28)S(18), E(18:2)O(29)S(18), and E(18:2)O(30)S(18) decreased by about 3% or more compared to before consuming the sample in the experimental group, and the relatively long-chain E(18:2)O(31)S(18), E(18:2)O(32)S(18) increased by about 3% or more, preferably 5% or more. Considering the statistical significance level, the proportion of relatively long-chain 32:0 ceramide increased, and relatively short-chain 28:0 ceramide and 30:0 ceramide decreased. Since skin itching, sleep disturbance, and atopic dermatitis increased at that time, it is interpreted that the ceramide skin lipid result affected atopic dermatitis (skin improvement).

[0093] Decreased synthesis of ceramide, a key component of skin barrier lipids, leads to impaired lamellae formation, lipid lamellae structure formation, and overall decreased epidermal lipids, resulting in impaired skin barrier function. EOS ceramides play a crucial role in the pathophysiology of atopic dermatitis due to their high hydrophobicity, and the length of their fatty acid chains is crucial for maintaining skin barrier function. Shorter fatty acid chains in atopic skin increase skin barrier permeability, disrupting skin barrier function and exacerbating skin symptoms.

[0094] Therefore, in this study, it can be seen that the beneficial effect was shown by reducing moisture loss in the skin through an increase in 32:0-EOS ceramide, a relatively long-chain fatty acid, and a decrease in 28:0-EOS ceramide and 30:0-EOS ceramide, which are relatively short-chain fatty acids, after administering kestos.

[0095]

[0096] Example 3: Skin microbiota analysis (skin smear test)

[0097] (1) Microbial flora analysis

[0098] On visits 2 and 4, skin smear tests were performed on the lesioned and non-lesioned areas (right anterior orbital fossa). First, a 200 μL sterile saline solution was placed in a container, and a cotton swab was dipped and removed. Microorganisms were collected by rubbing the lesioned and non-lesioned areas (right anterior orbital fossa) 1 cm away from the site where skin barrier function was measured 20–30 times with the soaked cotton swab. The sampled cotton swabs were stored frozen at -80°C in 0.9% saline solution until analysis.

[0099] For the obtained skin specimens, DNA was extracted using the Maxwell RSC PureFood GMA and Authentication Kit (Promega), amplified by PCR with fusion primers (341F and 805R) targeting the V3-V4 region of the bacterial 16S rRNA gene, and sequenced using the Illumina MiSeq Sequencing system (Illumina, USA). 16S rRNA gene sequence data were analyzed using QIIME2 software.

[0100] Skin microbiome analysis and protein analysis were performed on 48 subjects with compliance rates of 80%-120% in the order of randomization number.

[0101]

[0102] (2) Results of skin microbiome diversity analysis

[0103] To confirm the species diversity and profiling of the skin microbiome, skin samples obtained before and after 12 weeks of Kestos intake were analyzed for species diversity and profiling of the skin microbiome.

[0104] The species diversity and profiling analysis results of the skin microbiome are shown in Fig. 2, and Fig. 2 is a graph analyzing the skin microbial alpha diversity Chao1 index according to the intake of kestos.

[0105] In the skin microbiota alpha diversity Chao1 index analysis results of the above Fig. 2, Group A is the test result using a test sample containing Kestose powder as the main ingredient, Pre_A is the result before consuming the test sample, and Post_A is the result after consuming the test sample. Group B is the test result using a control sample containing Maltose powder as the main ingredient, Pre_B is the result before consuming the control sample, and Post_B is the result after consuming the control sample.

[0106] The results of the comparative analysis between each test group and the control group according to the results of the above Figure 2 are shown in Table 3 below, and it was confirmed that the alpha diversity Chao1 diversity index significantly increased after consuming Kestose.

[0107] Analysis details Statistical analysis results 1) Between-group analysis before administration between Group A and Group B (Pre_A vs Pre_B) No significant difference 2) Between-group analysis after administration between Group A and Group B (Post_A vs Post_B) 3) Within-group analysis before and after administration in Group A (Pre_A vs Post A) Chao1 (p=0.02) 4) Within-group analysis before and after administration in Group B (Pre_B vs Post_B) No significant difference

[0108] (3) Results of skin microbiome analysis

[0109] The microbiome can be designated by taxonomy, which organizes the lineage and affiliation of organisms based on specific criteria. The genus level was confirmed for the samples before and after ingestion of Kestos by group. The results of the skin microbiome flora analysis are shown in Fig. 3. Fig. 3 shows the results of analyzing the changes in the skin microbiome community according to Kestos ingestion at the genus level, and comparing the relative abundance ratio of the entire microorganisms as 100%, and shows the results of analyzing the Prevotella genus microorganisms among the skin microorganisms for the groups that ingested the control sample and the test sample, respectively, and shows the relative abundance ratio (%) of the Prevotella genus microorganisms.

[0110] In the results of the analysis of Prevotella genus microorganisms among skin microorganisms according to the intake of Kestose in the above Fig. 3, Group A is the test result using a test sample with Kestose powder as the main ingredient, Pre_A is the result before consuming the test sample, and Post_A is the result after consuming the test sample. Group B is the test result using a control sample with Maltose powder as the main ingredient, Pre_B is the result before consuming the control sample, and Post_B is the result after consuming the control sample. Specifically, in the results of the within-group analysis before and after administration in Group A, (Pre_A vs Post_A) g__Prevotella (p=0.007) was obtained, confirming that the Prevotella genus microorganisms significantly increased after consuming Kestose.

[0111] According to the comparative analysis results between each test group and the control group according to the results of Figure 3 above, skin microbiota profiling was confirmed through taxonomic composition at the genus level. At the Prevotella genus level, a significant increase was confirmed in the ratio from before (3.00%±0.02) to after (4.76%±0.03) intake of Kestos.

[0112]

[0113] Example 4: Analysis of the ratio of long-chain / short-chain fatty acids in skin lipids

[0114] (1) Sleep disturbance analysis

[0115] The following experiment was conducted to confirm the correlation between the improvement of skin disease symptoms due to Kestos and the ratio of EOS long-chain / short-chain fatty acids, using substantially the same method as in Example 1, including sample preparation, subject selection, and sample intake. Specifically, sleep disturbance scores were evaluated before and after 12 weeks of intake of the main ingredient Kestos as the test sample.

[0116] Specifically, the sleep disturbance score measurement method involves having the subject rate the severity of their sleep disturbance symptoms on a scale of 0 to 10. However, if the subject had difficulty assessing the severity of their symptoms, the subject's parent (or legal representative) performed the assessment on their behalf. The results of the sleep disturbance score measurement are shown in Figure 4.

[0117] As a result, after consuming the Kestos sample for 12 weeks, the average sleep disturbance score decreased from 4.08±2.41 to 2.17±1.58 after consuming Kestos, which was a decrease of -1.92±2.39 (P=0.0007).

[0118]

[0119] (2) Analysis of peripheral blood eosinophil count

[0120] In addition, the proportion of eosinophils among peripheral blood cells, which increase in cases of atopic disease because they play a role in protecting the body from external pathogens or infections, significantly decreased after taking kestose.

[0121] It was confirmed that the number of eosinophils in the peripheral blood increases when suffering from allergies, atopy, or other diseases, as they play a role in protecting the body from external pathogens or infections.

[0122] For analysis of peripheral blood eosinophil counts, blood was collected and analyzed from the subjects at Visits 1 and 4. Specifically, 1 cc of blood (EDTA tube) was collected for the immune-related blood eosinophil count, and 5 cc of blood (SST) was collected for the ECP and Total IgE tests. These anonymized samples were analyzed at another institution (Ewon Medical Foundation). The results of the analysis of peripheral blood eosinophil counts according to the above Kestos intake are shown in Figure 5.

[0123] As shown in Figure 5, the proportion of eosinophils among peripheral blood cells was significantly reduced after ingestion of kestose compared to before ingestion of kestose.

[0124]

[0125] (3) EOS ceramide ratio analysis

[0126] The skin barrier lipids were analyzed in the same manner as in Example 2 above, and the ratio analysis method corresponds to the fraction calculated as each EOS ceramide concentration * 100 / total ceramide concentration as described in [Table 2] of Example 2 above.

[0127] Figures 6 to 8 show the results of evaluating the relationship between 28:0-EOS ceramide and IGA (Investigator's Global Assessment), the relationship between 30:0-EOS ceramide and skin itching score, and the relationship between the ratio of 30:0-EOS ceramide and 32:0-EOS ceramide and skin itching score.

[0128] We confirmed that 30:0-EOS ceramide, a relatively short-chain fatty acid, had a positive correlation with the Investigator's Global Assessment (IGA), a global assessment scale for atopic dermatitis, and pruritus symptoms. Furthermore, we confirmed that pruritus symptoms increased as the proportion of the short-chain fatty acid 30:0-EOS ceramide increased and the proportion of the long-chain fatty acid 32:0-EOS ceramide decreased.

[0129] Through this, it was confirmed that the result of relatively increasing the skin long-chain fatty acid 32:0-EOS ceramide and relatively decreasing the skin short-chain fatty acids 28:0-EOS ceramide and 30:0-EOS ceramide after 12 weeks of Kestose intake also affected the relief of skin itching and skin disease symptoms. Figure 8 is a graph confirming the ratio of E(18:2)O(30)S(18) / E(18:2)O(32)S(18) and Itching. Since the correlation coefficient (r) has a positive value and the p-value is less than 0.05, it is statistically significant. Specifically, as 30:0-EOS ceramide increases and 32:0-EOS ceramide decreases, the itching value increases, so it was confirmed that the skin lipid improved according to the EOS short-chain / long-chain fatty acid ratio in the Kestose intake group.

Claims

1. A composition for controlling the content ratio of EOS ceramide in the stratum corneum of the skin, containing kestos as an active ingredient.

2. In the first paragraph, the composition increases the content ratio of at least one EOS ceramide selected from the group consisting of EOS ceramide (d18:1 / 31:0 / 18:2) and EOS ceramide (d18:1 / 32:0 / 18:2) of the stratum corneum of the skin in the total EOS ceramide.

3. In the second paragraph, the composition reduces the content ratio of at least one EOS ceramide selected from the group consisting of EOS ceramide (d18:1 / 28:0 / 18:2) and EOS ceramide (d18:1 / 30:0 / 18:2) in the total EOS ceramide of the stratum corneum.

4. In the first paragraph, the composition is a composition that reduces the content ratio of at least one EOS ceramide selected from the group consisting of EOS ceramide (d18:1 / 28:0 / 18:2) and EOS ceramide (d18:1 / 30:0 / 18:2) in the total EOS ceramide of the stratum corneum.

5. A composition according to any one of claims 1 to 4, wherein the composition is a pharmaceutical composition, a food composition, or a cosmetic composition.

6. A composition according to any one of claims 1 to 4, wherein the kestose is provided as a mixture with at least one sugar selected from the group consisting of nystose (GF3) and 1-F-fructosyl nystose (GF4).

7. In the 6th paragraph, when the kestose is provided as a mixture with one or more sugars selected from the group consisting of nystose (GF3) and 1-F-fructosyl nystose (GF4), the composition has a kestose content of 50 wt% or more based on 100 wt% of the total mixture.

8. A composition according to any one of claims 1 to 4, wherein the kestos is included in the composition in a total daily dosage of 1 g to 30 g based on a 60 kg adult.

9. A composition according to claim 1, wherein the kestose is provided in the form of a syrup or powder containing kestose.

10. A composition according to any one of claims 1 to 4, wherein the composition is for improving skin barrier lipids or improving skin microbiota, or preventing, treating, or improving diseases related to skin lipids or skin microbiota, or improving skin conditions.

11. A composition for increasing the relative proportion of Prevotella microorganisms in the skin microbiota, comprising kestos as an active ingredient.

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