Skin topical composition

Abstract

The present invention relates to a skin topical composition which is harmless to the human body and can fundamentally restore a skin barrier function. The composition comprises, as an active ingredient, a composite compound including: an Al-BA compound where lginate is conjugated with phenylboronic acid; and bacterial cellulose nanofibers ((+)BCNF) with positively charged surfaces. The skin topical composition of the present invention is harmless to the human body, can fundamentally restore the skin barrier function, and can be formulated into cosmetics, quasi-drugs, or pharmaceutical preparations.

Description

Composition for external skin application

[0001] The present invention is a composition for external application to the skin that is harmless to the human body and can fundamentally restore the skin barrier function.

[0002] The skin protects the body from physical and chemical stimuli of the external environment, while also playing a vital role in maintaining moisture and body temperature. From the outside, the skin is divided into three layers: the epidermis, the dermis, and the subcutaneous tissue. The epidermis is further divided into the stratum corneum, the granular layer, the spinous layer, and the basal layer, in that order.

[0003] Among these, the stratum corneum is located at the outermost part of the skin and is composed of about 58% keratin protein, about 38% natural moisturizing factor (NMF) that plays a role in retaining moisture, and about 11% lipids, and functions as a skin barrier. In other words, the stratum corneum is composed of protein-rich keratinocytes and intercellular lipids made up of various lipids such as ceramide, free fatty acids, and cholesterol. By forming a multilayer structure with keratinocytes and the lipids between them, it forms a complete skin barrier. The main function of this skin barrier is to prevent loss of body fluids and block attacks from toxins or invasion from pathogens.

[0004] However, the skin barrier is frequently damaged due to genetic or environmental factors, and this damaged skin barrier not only causes simple mechanical damage but also serious immune response problems through the penetration of various external toxins and pathogens. In particular, the increase in the number of people who enjoy sunbathing for cosmetic purposes has increased the number of cases of exposure to strong UV rays. In addition, the incidence of diseases related to skin damage is on the rise due to the inflammatory response caused by the penetration of skin toxins such as fine dust and the increase in moisture loss from the skin due to the weakened barrier function. Furthermore, when the skin barrier function is damaged, the moisture content of keratinocytes decreases, making the skin rough. In more severe cases, keratosis, the sloughing of keratinocytes, occurs. Furthermore, external toxins or microorganisms that penetrate the weakened skin barrier can trigger an excessive immune response within the skin, leading to hyperkeratosis and related diseases (e.g., atopy, psoriasis, etc.).

[0005] Conventionally, to address this skin barrier damage problem, methods have been used to supplement the skin with moisturizing factors (e.g., hyaluronic acid, ceramides, etc.). A representative example of this related prior art is Korean Patent No. 2366360 (Patent Document 1; KR10-2366360 B1). The entire contents of Patent Document 1 are hereby incorporated herein by reference as prior art.

[0006] [Prior Art Literature]

[0007] [Patent Document]

[0008] (Patent Document 1) KR10-2366360B1 (February 18, 2022)

[0009] As mentioned above, conventional techniques to solve the problem of skin barrier damage have mainly taken the approach of supplementing moisturizing factors, but the moisturizing factor supplementation method has the problem that the moisturizing power decreases over time and the skin condition returns to its original state, and it is not a method that fundamentally restores the skin barrier, so it is only a temporary expedient.

[0010] Accordingly, the present invention aims to provide a cosmetic, over-the-counter drug, or pharmaceutical composition that is harmless to the human body and can fundamentally restore the skin barrier function.

[0011] The present invention has been devised to solve the problems of the prior art,

[0012] A composition for external application to the skin is provided, which comprises as an active ingredient a complex compound of an Al-BA compound in which phenylboronic acid is combined with alginate and a bacterial cellulose nanofiber ((+)BCNF) having a positively charged surface.

[0013] In addition, in the present invention, a skin external application composition is provided, characterized in that the complex compound of the Al-BA compound and (+)BCNF is included in an amount of 0.1 to 10 wt% based on the total weight of the composition.

[0014] In addition, in the present invention, a skin external application composition is provided, characterized in that the weight ratio of Al-BA complex compound: bacterial cellulose nanofiber (BCNF) = 1: 0.01 to 0.5.

[0015] In addition, the present invention provides a skin external application composition further comprising one or more moisturizing or skin barrier components selected from glycerin, hyaluronic acid, and cholesterol.

[0016] In addition, in the present invention, a skin external application composition is provided, characterized in that the composition is a cosmetic composition for restoring skin barrier function and promoting skin moisturizing.

[0017] In addition, in the present invention, a skin external application composition is provided, characterized in that the composition is a pharmaceutical composition for protecting the skin in an area where the skin barrier is damaged.

[0018] In addition, in the present invention, a skin external application composition is provided, characterized in that the above-mentioned drug is an adhesive wound dressing medical device.

[0019] In addition, in the present invention, a composition for external application to the skin is provided, characterized in that the composition is a pharmaceutical composition for treating, preventing, or alleviating one or more diseases or symptoms selected from the group consisting of dry skin, pruritus, dry eczema, hyperkeratosis of the skin, keratosis of the heel and ankle area, xeroderma in the elderly, rough skin in adults, actinic dermatitis, atopic dermatitis, and psoriasis.

[0020] The present invention is a composition for external application to the skin, which is harmless to the human body and can fundamentally restore the skin barrier function, and can be manufactured into a cosmetic, an over-the-counter drug, or a pharmaceutical composition.

[0021] Figure 1 shows (a) a schematic diagram of Alginate-Boronic acid (Al-BA) conjugation, (b) FT-IR analysis of Al-BA synthesis, and (c) quantification of conjugated boronic acid.

[0022] Figure 2 shows (a) a schematic diagram of the synthesis of positively charged natural cellulose ((+)BCNF), (b) surface potential values ​​of natural cellulose according to the amount of glycidyltrimethylamine chloride (GTAC) introduced, and (c) a TEM image of (+)BCNF.

[0023] Figure 3 is a graph showing the results of measuring the recovery of skin barrier damage caused by chemical stimulation.

[0024] Figure 4 is a graph showing the results of measuring skin barrier damage recovery due to optical stimulation.

[0025] Figures 5 and 6 are graphs showing the results of measuring the recovery of skin damage caused by physical stimulation.

[0026] Figure 7 is a graph of the results of each mass measurement.

[0027] Figure 8 is a graph of moisture content measurement results.

[0028] Figure 9 is a graph showing the results of measuring transepidermal water loss.

[0029] Figure 10 is a graph showing the results of skin roughness (skin texture) measurement.

[0030] Figure 11 is a graph showing the results of visual evaluation of cracking.

[0031] Figure 12 is a graph showing the results of measuring the moisture content of the forearm skin by concentration and time.

[0032] Figure 13 is a schematic diagram of the overall schedule and scheme for the atopic dermatitis efficacy evaluation test.

[0033] Figures 14 to 17 are graphs showing the results of tests on the skin of atopic dermatitis-induced areas for symptoms of erythema / hemorrhage, scaling / dryness, edema, and skin peeling / erosion.

[0034] Figure 18 is a graph showing the results of measuring the thickness of the outer skin of atopic dermatitis-induced skin.

[0035] Figures 19 to 24 are photographs of the results of visual inspection and histopathological examination.

[0036] First, the terms used in the present invention are defined.

[0037] The term "skin" in the present invention refers to the tissue covering the body surface of an animal, and is a broad concept that includes not only tissue covering the body surface such as the face or body, but also the scalp and hair. Meanwhile, in the present specification, "skin" may include not only biological skin, but also artificial skin or a skin mimic that embodies the state of biological skin.

[0038] In the present invention, the term "skin barrier function" refers to all functions of the skin, particularly the stratum corneum of the epidermis, which serve as a barrier layer against the outside, such as preventing moisture from leaking out and controlling the entry and exit of substances from the outside, but is not limited thereto.

[0039] The term "skin barrier dysfunction" as used herein refers to not only a state in which the skin barrier function is reduced or damaged, but also any state in which the skin barrier function is likely to be reduced or damaged, and any condition requiring prevention thereof, and is not limited by the type or degree of specific symptoms. More specifically, the skin barrier dysfunction may include, but is not limited to, inflammatory reactions caused by external stimuli such as ultraviolet rays and fine dust, and weakening of the epidermis due to aging.

[0040] The term "improvement" in the present invention refers to, but is not limited to, any action that removes the skin barrier function from a state of deterioration or damage, a state at risk of damage, or a state requiring prevention. More specifically, the improvement may include, but is not limited to, an action that induces epidermal differentiation and promotes epidermal turnover by activating the autophagy of epidermal keratinocytes, which are reduced by external stimuli or aging, thereby ultimately restoring the epidermal barrier function and preventing aging.

[0041] The term "treatment" as used herein means any act of improving or completely curing the symptoms of a disease by administering a composition according to the present invention.

[0042] The term “prevention” as used herein means any act of suppressing or delaying the symptoms of a disease by administering a composition according to the present invention.

[0043] As used herein, the term "alleviation" means any action by which the symptoms of a disease are at least reduced or benefited by administration of the pharmaceutical composition according to the present invention.

[0044]

[0045] Hereinafter, the present invention will be described in detail.

[0046] The present invention is a composition for external application to the skin comprising, as an active ingredient, a complex compound of an Al-BA compound in which phenylboronic acid is bonded to alginate and a bacterial cellulose nanofiber ((+)BCNF) having a positively charged surface.

[0047] The above Al-BA compound is a complex compound in the form of conjugated phenylboronic acid to alginate, and is capable of inducing adhesion to the skin through rapid sol-gel transition in a biological pH environment.

[0048] The complex compound of the above Al-BA compound and (+)BCNF may be included in an amount of 0.1 to 10 wt%, and preferably 1 to 5 wt%, based on the total weight of the composition. Below the above range, the intended effect on the skin barrier may be insufficient, and above the above range, there may be concerns about problems such as decreased formulation stability due to increased viscosity, decreased ease of handling, and decreased uniformity of dispersibility of the formulation.

[0049] The above bacterial cellulose nanofibers not only increase skin adhesion, but also contribute to the formation of a skin protective layer and the recovery of damaged skin tissue. That is, due to their structure similar to the extracellular matrix, they can accelerate the proliferation rate of skin cells in damaged or wounded areas. In particular, the bacterial cellulose nanofibers preferably have a positively charged surface to enhance their affinity for the Al-BA compound and skin tissue, and to improve their physical properties.

[0050] The content of the above Al-BA complex compound is 1 to 10 wt% relative to the total weight of the composition,

[0051] The content of the above bacterial cellulose nanofibers (BCNF) may be included in an amount of 1 wt% or less, preferably in a range of 0.01 to 0.5 wt%, based on the total weight of the composition.

[0052] In the present invention, it is preferable to consider the ratio of the Al-BA compound and bacterial cellulose nanofibers (BCNF). That is, the weight ratio of Al-BA complex compound: bacterial cellulose nanofibers (BCNF) may be within the range of 1:0.01 to 0.5, preferably 1:0.02 to 0.2, and more preferably 1:0.03 to 0.16. This is because a weight ratio within this range exhibits more desirable properties for forming a skin protective layer, and if this range is exceeded, there may be a concern that the polymers may be unevenly dispersed due to the viscosity difference between the Al-BA compound and BCNF, thereby inhibiting the performance of forming a protective layer by adhering to the skin. A composite compound of Al-BA / (+)BCNF, in which 0.25 wt% (+)BCNF is introduced into approximately 3 wt% Al-BA complex compound, can be preferably used as the optimal concentration ratio.

[0053] The composition of the present invention may further include active ingredients, such as conventionally well-known moisturizing ingredients such as glycerin and hyaluronic acid, and conventionally well-known skin barrier components such as cholesterol, which acts as mortar for the skin barrier in the so-called "Brick & Mortal Model," within a range that does not impair the effects of the present invention, and is not particularly limited thereto. In addition, it goes without saying that the composition may further include conventional auxiliary agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances, and carriers.

[0054]

[0055] The skin external application composition of the present invention can be provided in any form of a cosmetic, over-the-counter drug, or pharmaceutical composition that can restore skin barrier function.

[0056] According to one embodiment, the skin external application composition may be manufactured as a cosmetic composition. Even when used routinely, it can provide skin protection and damaged skin recovery effects, and can continuously improve the skin barrier function without causing skin trouble, thereby providing an effect of improving the overall skin condition. The cosmetic composition according to one embodiment of the present invention is not limited to being manufactured in any formulation commonly manufactured in the art, and may be formulated in any form, for example, a solution, a gel, a solid, a paste, an anhydrous product, an emulsion, a suspension, an emulsion, a microemulsion, a microcapsule, a microgranule, a liposome, a non-ionic vesicle dispersion, a serum, a cream, a skin, a lotion, a powder, a spray, a paste, a pack, a cleanser, a soap, an oil, a wax, a concealer stick, or an aerosol, but is not limited thereto. More specifically, it can be applied to manufacturing functional cosmetics, which are basic cosmetics such as toner, nourishing cream, moisturizing cream, essence, eye cream, lotion, pack, and essence.

[0057] The content of the extract included as an active ingredient in the cosmetic composition according to the present invention may be appropriately determined by considering various factors such as the purpose of use, period of use, type of formulation, route of administration, and skin condition of the subject. For example, the content of the extract may be 0.0001 to 10 wt%, for example, 0.001 to 5 wt%, based on the total weight of the cosmetic composition.

[0058] According to another embodiment, the present invention can be used as a quasi-drug composition for improving skin barrier function. "Quasi-drug" refers to products that are used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases in humans or animals, and have a milder effect than a pharmaceutical product. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are products other than those used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases in humans or animals, products that have a mild effect or do not directly affect the human body, etc.

[0059] Like the cosmetic composition described above, the above-mentioned non-medical product composition can be used for the purpose of enhancing skin moisturization and restoring damaged skin barrier, but it can also have a purpose of protecting the skin in the damaged skin barrier area, and can also be used as a medical device such as an adhesive (transparent) wound dressing for covering and protecting skin wounds caused by various causes.

[0060] The above-described quasi-drug composition of the present invention may be exemplified as a disinfectant, a shower foam, an ointment, a wet tissue, a coating agent, etc., and may preferably be manufactured as a semi-solid preparation such as an external ointment, lotion, etc., but is not limited thereto. The formulation method, dosage, method of use, components, etc. of the above-described quasi-drug may be appropriately selected from conventional techniques known in the art.

[0061] The content of the extract included as an active ingredient in the quasi-drug composition according to the present invention may be appropriately determined by considering various factors such as the purpose of use, period of use, type of formulation, application route, and skin condition of the subject. For example, the content of the extract may be 00001 to 10 wt%, for example 0001 to 5 wt%, based on the total weight of the quasi-drug composition.

[0062] According to another embodiment, it can be used as a pharmaceutical composition for treating, preventing, or alleviating diseases related to skin barrier damage. Diseases related to skin barrier damage include various infectious and inflammatory diseases that damage the skin barrier or are caused by skin barrier damage, and specific examples thereof include dry skin, pruritus, dry eczema, hyperkeratosis of the skin, keratosis of the heel and ankle area, xeroderma in the elderly, rough skin in adults, actinic dermatitis, atopic dermatitis, and psoriasis.

[0063] The pharmaceutical composition of the present invention, the skin external application composition of the present invention, can be manufactured in any formulation commonly manufactured in the art, for example, can be manufactured in the form of a solution, gel, solid, paste, anhydrous product, emulsion, suspension, emulsion, microemulsion, microcapsule, microgranule, liposome, nonionic vesicle dispersion, serum, cream, skin, lotion, powder, spray, paste, pack, cleanser, soap, oil, wax, concealer stick or aerosol.

[0064]

[0065]

[0066] Hereinafter, the present invention will be described in more detail using examples and test examples. However, it should be made clear that the following examples are intended only to provide a detailed explanation of the invention and are not intended to limit the scope of the rights herein.

[0067]

[0068] Example

[0069] Preparation of a composite compound of Al-BA / (+)BCNF

[0070] Synthesis of Al-BA Compound: First, alginate polymer was dispersed in MES buffer (0.1 M, pH 5.0) at room temperature. A mixed solution of ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) / N-hydroxysuccinimide (NHS) was slowly added to the alginate dispersion while stirring. Then, a solution of 3-aminophenylboronic acid was also slowly added and stirred for 24 hours. After the reaction was completed, the residue was removed through dialysis to produce alginate-boronic acid (Al-BA). The prepared Al-BA was freeze-dried and stored.

[0071] Identification of Al-BA compound: When FT-IR analysis was performed before and after introduction of 3-aminophenylboronic acid into alginate polymer, 3-aminophenylboronic acid and 3000-2800 cm -1 and 1650-1580 cm -1 Synthesis was confirmed by observing similar peaks in both regions. Furthermore, the introduced 3-aminophenylboronic acid (BA) was quantified through UV-Vis analysis. The results are shown in Figure 1.

[0072] Synthesis of BCNF: Meanwhile, natural cellulose raw material was dispersed in a 0.1 M NaOH solution, and a quantitative amount of glycidyltrimethylamine chloride was added. The mixture was stirred at 60°C for 2 hours. The residue was then removed by centrifugation, and the reaction product was redispersed in a 0.01 M NaOH solution. NH4OH was added to the cellulose redispersion, stirred at 60°C for 2 hours, and centrifuged to obtain positively charged natural cellulose nanofibers ((+)BCNF).

[0073] Confirmation of BCNF: Analysis of the surface potential according to the amount of glycidyltrimethylamine chloride (GTAC) functional groups introduced onto the surface of natural cellulose nanofibers revealed that the surface potential increased in a positive range as the amount of GTAC introduced increased. Furthermore, the nanofiber structure of the manufactured (+) BCNF was visualized using TEM imaging. The results are shown in Fig. 2.

[0074] Preparation of a complex compound of Al-BA / (+)BCNF: The Al-BA prepared above and the (+)BCNF obtained above were added to water at a concentration ratio of 3 wt% and 0.25 wt%, respectively, and mixed to finally prepare a complex compound of Al-BA(+)BCNF in the form of a hydrogel.

[0075]

[0076] Using the complex compound of Al-BA / (+)BCNF manufactured as described above as an effective ingredient, compositions of Examples 1 and 2 and Comparative Examples were manufactured as prescribed in Table 1 below (unit: weight %).

[0077] Ingredient name Comparative example Example 1 Example 2 Purified water TO.100 TO.100 TO.100 Glycerin 5.0000 5.0000 5.0000 Sodium polyacrylate 0.30000 30000 3000 Xanthan gum 0.05000 05000 0500 Cetyl ethyl hexanoate 8.0000 8.0000 8.0000 Hydrogenated polydecene 4.0000 4.00004.0000 Glyceryl Stearate 3.04003.04003.0400 Cetearyl Alcohol 3.00003.00003.0000 Cholesterol 2.86802.86802.8680 Dimethicone 2.00002.00002.0000 Hydroxypropyl Bispalmitamide MEA 2.00002.0 0002.0000 PEG Stearate 0.96000.96000.9600 Glyceryl Caprylate 0.50000.50000.5000 Candelilla Wax 0.50000.50000.5000 Palmitic Acid 0.05460.05460.0546 Stearic Acid 0.04420.04420.0442 Lauric Acid 0.0006 0.0006 0.0006 Myristic Acid 0.0006 0.0006 0.0006 Purified Water 2.0000 2.0000 2.0000 Arginine 0.20000 20000 2000 Al-BA / (+)BCNF-0.5000 1.0000 1,2-Hexanediol 1.0000 1.0000 1.0000

[0078]

[0079] Safety and efficacy verification tests

[0080] To confirm the efficacy and safety of the formulation using Al-BA / (+)BCNF and to confirm various effects such as skin barrier recovery and skin moisturizing enhancement, clinical trials were conducted on various items as follows.

[0081]

[0082] 1. Subject information and test product

[0083] There were a total of 11 clinical trial subjects, and the general information was as shown in Table 2 below.

[0084]

[0085] Meanwhile, the test product used the above-mentioned manufactured Example 2, and the control product used the above-mentioned manufactured Comparative Example.

[0086]

[0087] 2. Measurement of skin barrier damage recovery caused by chemical stimulation

[0088] [Test method]

[0089] To induce skin barrier damage by chemical stimulation, the test area was marked on the left forearm of the test subject without pigmentation or skin damage, and skin damage was induced by applying a 2% SLS (Sodium Lauryl Sulfate) solution as an occlusive patch for 24 hours using an IQ Ultimate chamber (Chemotechnique Diagnostics, Sweden).

[0090] Skin barrier recovery from chemical irritation was measured using a Vapometer (Delfin Technologies Ltd, Finland) on the same left forearm before SLS stimulation, 24 hours after SLS stimulation, and 2 weeks after use of the test product. The unit of measurement was g / ㎡h, and a decrease in the measured value indicates a greater effect on skin barrier recovery from chemical irritation.

[0091] [Test Results]

[0092] The results of the skin barrier damage recovery measurement due to chemical stimulation (unit: g / ㎡h) were as shown in Table 3 and Figure 3 below.

[0093]

[0094] - Area of ​​use of test product: Statistically significant increase was observed 24 hours after SLS stimulation compared to before SLS stimulation, and statistically significant improvement was observed 2 weeks after use compared to 24 hours after SLS stimulation.

[0095] - Areas where control product was used: Statistically significant increase was observed 24 hours after SLS stimulation compared to before SLS stimulation, and statistically significant improvement was observed 2 weeks after use compared to 24 hours after SLS stimulation.

[0096] - Comparison between groups: There was no statistically significant difference 24 hours after SLS stimulation compared to before SLS stimulation, but a statistically significant difference was shown 2 weeks after use compared to 24 hours after SLS stimulation.

[0097]

[0098] 3. Measurement of skin barrier damage recovery due to optical stimulation

[0099] [Test method]

[0100] To induce skin barrier damage by optical stimulation, UV light was irradiated using a Multiport Solar Simulator, 601-300W (Solar Light, USA). The test area was marked on the right forearm of the test subject without pigmentation or skin damage, and UV light of the same intensity (650 ㎼ / ㎠) was irradiated for 1 minute 30 seconds (±10 seconds).

[0101] Measurements of skin barrier damage recovery induced by optical stimulation were performed using a Vapometer (Delfin Technologies Ltd, Finland). Measurements were taken on the same right forearm before UV exposure, 24 hours after UV exposure, and 2 weeks after use of the test product. The unit of measurement was g / ㎡h, and a decrease in the measured value indicates a greater effect on skin barrier damage recovery induced by optical stimulation.

[0102] [Test Results]

[0103] The results of the measurement of skin barrier damage recovery due to optical stimulation (unit: g / ㎡h) were as shown in Table 4 and Figure 4 below.

[0104]

[0105] - Area of ​​use of the test product: Statistically significantly increased 24 hours after UV exposure compared to before UV exposure, and statistically significantly improved 2 weeks after use compared to 24 hours after UV exposure.

[0106] - Area of ​​use of control product: Statistically significant increase 24 hours after UV irradiation compared to before UV irradiation.

[0107] - Comparison between groups: There was no statistically significant difference 24 hours after UV exposure compared to before UV exposure, and a statistically significant difference was observed 2 weeks after use compared to 24 hours after UV exposure.

[0108]

[0109] 4. Measurement of skin barrier damage recovery due to physical stimulation

[0110] [Test method]

[0111] To induce skin barrier damage by physical stimulation, the test area was marked on the left forearm of the test subject without pigmentation or skin damage, and skin damage was induced (TS: Tape Stripping) by repeatedly attaching and removing a film (D-Squame; Cuderm, USA).

[0112] Measurements of skin barrier damage recovery by physical stimulation were performed using a Vapometer (Delfin Technologies Ltd, Finland) on the same left forearm before, after, and immediately after TS treatment. The measurement unit is g / ㎡h, and a decrease in the measured value indicates a greater effect on skin barrier damage recovery by optical stimulation.

[0113] Additionally, the same left forearm was measured before, after, and immediately after TS using Antera 3D CS (Miravex Ltd., Ireland). Analysis was performed by designating specific areas of each segmented area from the captured images (color mode). The parameter is the a* value, which indicates skin redness. A decrease in the a* value immediately after TS compared to after TS indicates an effect in recovering skin barrier damage caused by physical stimulation.

[0114] [Test Results]

[0115] The results of the skin barrier damage recovery measurement due to physical stimulation were as shown in Table 5 (unit: g / ㎡h), 6 (unit: a*) and Figures 5 and 6.

[0116]

[0117]

[0118] - Areas where the test product was used: Statistically significantly increased after TS compared to before TS, and statistically significantly improved immediately after use compared to after TS.

[0119] - Areas where control product was used: Statistically significantly increased after TS compared to before TS, and statistically significantly improved immediately after use compared to after TS.

[0120] - Comparison between groups: There was no statistically significant difference after TS compared to before TS, and a statistically significant difference was observed immediately after TS compared to after TS.

[0121]

[0122] 5. Measurement of each mass

[0123] [Test method]

[0124] For the measurement of keratin mass, a special film (D-Squame; Cuderm, USA) was used to collect keratin samples from the same heel area of ​​the test subjects on both sides before, immediately after, and two weeks after using the test product, and the samples were photographed with Visioscan VC20 (Courage+Khazaka eletronic GmbH, Germany). The analysis parameter was DI (Desquamation Index, %), and a decrease in the analysis value indicates a greater effect on improving keratin mass.

[0125] [Test Results]

[0126] The results of each mass measurement were as shown in Table 7 (unit: %) and Figure 7.

[0127]

[0128] - Areas where the test product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0129] - Areas where control product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0130] - Inter-group comparison: Statistically significant differences were observed immediately after use and 2 weeks after use compared to before use.

[0131]

[0132] 6. Moisture measurement

[0133] [Test method]

[0134] Moisture content was measured using a Corneometer CM825 (Courage+Khazaka eletronic GmbH, Germany) on the same heel area of ​​each subject before, immediately after, and two weeks after using the test product. Parameters are expressed in arbitrary units (AU), and higher values ​​indicate greater improvement in moisture content.

[0135] [Test Results]

[0136] The moisture content measurement results were as shown in Table 8 (unit: %) and Figure 8.

[0137]

[0138] - Areas where the test product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0139] - Areas where control product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0140] - Inter-group comparison: A statistically significant difference was observed 2 weeks after use compared to before use.

[0141]

[0142] 7. Measurement of transepidermal water loss

[0143] [Test method]

[0144] Transepidermal water loss (TEWL) was measured using a Vapometer (Delfin, Finland) on the same heel area of ​​each subject before, immediately after, and two weeks after use of the test product. The parameter was water loss (g / ㎡h), and a decrease in the analyzed value indicates a greater improvement in TWL.

[0145] [Test Results]

[0146] The moisture content measurement results were as shown in Table 9 below (unit: g / ㎡h) and Figure 9.

[0147]

[0148] - Areas where the test product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0149] - Areas where control product was used: No statistically significant improvement was observed compared to before use.

[0150] - Inter-group comparison: Statistically significant differences were observed immediately after use and 2 weeks after use compared to before use.

[0151]

[0152] 8. Skin roughness measurement

[0153] [Test method]

[0154] Skin roughness (texture) was measured using a Visioscan VC98 (Courage+Khazaka eletronic GmbH, Germany) on the same heel area of ​​each subject before, immediately after, and two weeks after using the test product. The parameter was R3 (Roughness), and a lower value indicates a greater improvement in skin roughness (texture).

[0155] [Test Results]

[0156] The results of skin roughness (skin texture) measurement were as shown in Table 10 (unit: R3) and Figure 10.

[0157]

[0158] - Areas where the test product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0159] - Areas where control product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0160] - Inter-group comparison: A statistically significant difference was observed 2 weeks after use compared to before use.

[0161]

[0162] 9. Visual measurement of cracks

[0163] [Test method]

[0164] Visual evaluation of cracking was conducted by two researchers before, immediately after, and two weeks after using the test product, referring to the paper "A new technique for evaluating heel xerosis grade and the effects of moisturizer on heel skin dryness."

[0165] A lower grade indicates a higher effectiveness in improving cracked heels, and the criteria in Table 11 below described in the paper were referenced.

[0166]

[0167] [Test Results]

[0168] The results of the splitting measurement were as shown in Table 12 (unit: grade) and Figure 11.

[0169]

[0170] - Areas where the test product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0171] - Areas where control product was used: Statistically significant improvement was observed immediately after use and 2 weeks after use compared to before use.

[0172] - Inter-group comparison: A statistically significant difference was observed 2 weeks after use compared to before use.

[0173]

[0174] 10. Validity Evaluation Survey Results

[0175]

[0176]

[0177]

[0178] 11. Product preference survey results

[0179]

[0180]

[0181] The skin external application composition of the present invention as described above was evaluated for recovery of skin barrier damage caused by chemical stimulation, recovery of skin barrier damage caused by optical stimulation, recovery of skin barrier damage caused by physical stimulation, keratin content, moisture content, transepidermal water loss, skin roughness (skin texture), and cracking, and it was confirmed to be significant, and no special skin adverse reactions, special toxicity, or side effects were observed during the period in which the test subject used the test product.

[0182]

[0183] Concentration dependence confirmation test

[0184] To confirm the concentration dependence, samples containing Al-BA / (+)BCNF were applied to the forearm area of ​​eight researchers, and the products of Comparative Example (0%), Example 1 (0.5%), and Example 2 (1.0%) were then applied. The skin moisture content was measured over time using a Corneometer (Cutometer® dual MPA 580). No particular abnormal symptoms were observed.

[0185] The results were as shown in Table 16 and Figure 12 below.

[0186] Before application 30 minutes after application 1 hour after application 2 hours after application Comparative Example 34.949.5340.9636.91 Example 134.954.3843.4738.7 Example 234.955.1944.5839.83

[0187]

[0188]

[0189] Atopic dermatitis efficacy evaluation

[0190]

[0191] 1. Test animals and breeding conditions

[0192] 1.1. Test animals

[0193] Species and strain: Female hairless mice (CriOri:SKH1-Hrhr) Reason for selecting the test system: The mice used in this test were selected because abundant basic test data has been accumulated, and the interpretation and evaluation of test results are easy using such data. Supplier and producer: Saeron Bio Co., Ltd. (251 Anyangpangyo-ro, Uiwang-si, Gyeonggi-do) Date of acquisition: April 26, 2023 Age at acquisition: 20 g ± 10% Weight range: 22 g ± 10% Quarantine and acclimatization: 7 days after acquisition (during the acclimatization period, general symptoms were observed to check the health status, and healthy animals were used in the test). Individual identification: Individuals are identified through tail tattoos, and individual identification cards recording the management number, population group, animal number, and sex are attached to each breeding room. Number of animals ordered: 36 Number of animals used: 36

[0194]

[0195] 1.2. Breeding conditions

[0196] Breeding Room: Inha University Hospital Digestive Disease Effectiveness Evaluation Support Center, Rodent Breeding Area, Room 1, Number of ventilations: 14-18 times per hour, Temperature and humidity range: 22±2℃, 50±10%, Lighting time and intensity: 12 hours (lights on 8:00 AM - lights off 8:00 PM), 150-300 Lux, Breeding box: Polysulfonate cage (Tree Shine, Korea) [200 W x 320 D x 145 H (mm)], Number of animals per breeding box: 6, Breeding management: Bedding was placed on the floor of the cage to absorb feces and urine, and the cage and bedding were replaced twice a week. Type and name of feed: Safe diet [Woojung Bio Co., Ltd.], Feed was supplied in a feeder and allowed to consume freely. Type of drinking water: Tap water sterilized with an ultraviolet sterilizer and a microfiltration device, Drinking water was supplied in a waterer and allowed to consume freely.

[0197]

[0198] 2. Test materials and methods

[0199] 2.1. Test substance

[0200] AL-BA / BCNF 0.3% Cream: A white cream-type adhesive transparent wound dressing containing 0.3 wt% of Al-BA / (+)BCNF as an active ingredient (solvent is purified water), and manufactured by adding thickeners, etc. to have a pH of 6.5 to 9 and a viscosity of approximately 30,000 to 60,000 cP (Brookfield Viscometer, RV type, Spindle No. 6, (10, 15, 20) r / min).

[0201]

[0202] 2.2. Control material

[0203] Estura Ato Barrier Itching Cream MD (manufactured by Amore Pacific) on sale

[0204] Appearance: White cream

[0205]

[0206] 2.3. Test method

[0207] The overall test group composition and types of test substances were as shown in Table 19 below.

[0208]

[0209] Induction of atopic dermatitis: Female mice weighing 20-24 g, acclimatized for more than 7 days, were subjected to general anesthesia using an inhalant anesthetic (0.5-2% isoflurane: Ifran® Hana Pharm Co Ltd) with oxygen, 1:1 (5-10 ml / kg / min)). 1% Oxazolone was administered (applied) to the skin around the scapula of the anesthetized mice and allowed to absorb. One week later, 0.3% Oxazolone was re-administered to induce atopy in the absorbed skin. Thereafter, 0.3% Oxazolone was administered once every two days to continuously induce atopy.

[0210] Test substance application: One minute after the third administration of 0.3% oxazolone, the test substance and control cream were applied to the atopic dermatitis-induced area. Administration was performed twice daily to the same area until the end of the study.

[0211] Autopsy: Autopsy was performed 14 days after administration of test and control substances.

[0212] The overall schedule and scheme for the test progress are shown in Figure 13.

[0213]

[0214] 3. Observation and inspection items

[0215] 3.1. Observation of general symptoms

[0216] During the test period, clinical symptoms and death / moribundity were observed for each individual animal. Records were maintained only for animals exhibiting specific clinical symptoms (normal animals were not recorded).

[0217]

[0218] 3.2. Atopy severity assessment

[0219] As shown in Table 20 below, the four items of erythema / hemorrhage, scaling / dryness, edema, and skin peeling / erosion were scored from 0 to 3 (normal: 0, mild: 1, moderate: 2, and severe: 3). The degree of atopy induction was evaluated by the sum of the scores.

[0220]

[0221] ※ The evaluation criteria were established by selecting the individual with the most severe symptoms among the individuals and setting the criteria as Severe: 3.

[0222]

[0223] 3.3. Autopsy

[0224] Ten days after administration of the test and control substances, the experimental animals were euthanized using carbon dioxide to terminate the experiment, and autopsies were performed. For histopathology, atopic skin tissue was excised and stored in a 10% formalin solution. Only organs with unusual findings were recorded and stored (normal organs were not recorded or stored).

[0225]

[0226] 3.4. Histopathological examination

[0227] The fixed tissues underwent general tissue processing such as trimming, dehydration, paraffin embedding, and sectioning to prepare specimens for histopathological examination, and then hematoxylin & eosin (H&E) staining was performed. The presence of administered substances and histopathological changes were observed using an optical microscope (Olympus BX53, Japan). Afterwards, the sections were scanned using a slide scanner (Carl Zeiss Axio Scan Z1, Germany), and the scanned images were analyzed using an image analysis program (Carl Zeiss ZEN, Germany).

[0228]

[0229] 3.5. Measurement of outer skin thickness

[0230] The histopathological images scanned using a slide scanner (Carl Zeiss Axio Scan Z1, Germany) were analyzed by measuring the 3-point epithelial thickness using Image J (Ver. 1.53f51).

[0231]

[0232] 4. Test results

[0233] 4.1. Death and general symptoms

[0234] During the study period, no deaths were observed due to atopic dermatitis or administration of the test or control substances. All subjects with atopic dermatitis experienced skin erythema / hemorrhage, scaling / dryness, edema, and skin peeling / erosion. However, no abnormal symptoms, such as fever or inflammation, were observed following administration of the test or control substances.

[0235]

[0236] 4.2. Degree of atopy

[0237] After the test period, the skin of all subjects in the atopic dermatitis-induced area was visually inspected and scored for symptoms such as erythema / bleeding, scaling / dryness, edema, and skin peeling / erosion.

[0238] The results were as shown in Tables 21 to 24 and Figures 14 to 17.

[0239]

[0240]

[0241]

[0242]

[0243] The mean scores for erythema / hemorrhage in each group were 2.25 ± 0.62 in the non-treated group (G1), 1.75 ± 1.06 in the test group (G2), and 2.00 ± 1.21 in the control group (G3), with no significant differences observed among the groups. The mean scores for scaling / dryness in each group were 2.58 ± 0.51 in the non-treated group (G1), 1.75 ± 0.62 in the test group (G2), and 2.00 ± 0.85 in the control group (G3), with a significant difference observed between the non-treated and test groups (G1 vs G2 p = 0.0134). No significant differences were observed between the non-treated and control groups. The mean score of edema by group was 2.50 ± 0.52 in the non-treated group (G1), 1.67 ± 0.78 in the test group (G2), and 1.92 ± 0.79 in the control group (G3). A significant difference was observed between the non-treated group and the test group (G1 vs G2 p = 0.0185). No significant difference was observed between the non-treated group and the control group. The mean score of skin lifting / erosion by group was 2.58 ± 0.51 in the non-treated group (G1), 1.83 ± 0.72 in the test group (G2), and 1.83 ± 0.72 in the control group (G3). A significant difference was observed between the non-treated group and the test group and between the non-treated group and the control group (G1 vs G2 p = 0.0227, G1 vs G3 p = 0.227). No significant difference was observed between the test group and the control group.

[0244]

[0245] 4.3. Thickness of the outer layer of atopic skin

[0246] After the test period, histopathological examinations were performed on the skin of all subjects at the atopic dermatitis-inducing sites. Tissue slides were stained with H&E, and epidermal thickness was measured and compared across groups.

[0247] The results were as shown in Table 25 and Figure 18 below.

[0248]

[0249] The thickness of the outer skin by group was 129.42 ± 36.27 ㎛ in the non-treated group (G1), 80.48 ± 20.91 ㎛ in the test group (G2), and 79.55 ± 26.20 ㎛ in the control group (G3). It was confirmed that there was a significant difference between the test group and the control group and the non-treated group (G1 vs G2 p = 0.0005, G1 vs G3 p = 0.0004). No significant difference was observed between the test group and the control group.

[0250]

[0251] 7. Discussion and Conclusion

[0252] Atopic dermatitis model induction and test substance administration (application) were performed on female hairless mice (CriOri:SKH1-Hrhr) acclimatized for more than 5 days. 1% Oxazolone was administered to the skin around the scapula and absorbed. Atopic dermatitis was induced one week later by re-administering 0.3% Oxazolone. Thereafter, 0.3% Oxazolone was administered once every two days to continuously induce atopic dermatitis. One minute after the third administration of 0.3% Oxazolone, the test substance and control substance cream were applied to the atopic dermatitis-induced area. Administration was performed twice daily to the same area until the end of the study. An autopsy was performed 10 days after administration of the test and control substances.

[0253] The group composition was G1: Non-treated group (Negative control: Non-treated), G2: Test group (Test: AL-BA / BCNF 0.3% cream applied), G3: Control group (Positive control: Estura Atobarrier Itching Cream applied), and the number of animals in each group was 12, for a total of 36 animals.

[0254] During the test period, no deaths due to atopic induction or test / control substance administration or specific clinical symptoms due to test / control substances were observed.

[0255] After the test period, all subjects were visually examined and scored for erythema / hemorrhage, scaling / dryness, edema, and skin peeling / erosion at the atopic dermatitis-induced and test substance-administered sites. No significant differences were observed in erythema / hemorrhage between the groups. Significant differences were observed in scaling / dryness and edema between the untreated and test groups, but no significant differences were observed between the untreated and control groups. Significant differences were observed in skin peeling / erosion between the untreated and treated groups (test and control groups), but no significant differences were observed between the test and control groups.

[0256] Significant differences in skin thickness were observed between the non-treated and test groups and between the non-treated control group, but no significant differences were observed between the test and control groups.

[0257] The test results confirmed that the test and control substances significantly alleviated the symptoms of atopic dermatitis and significantly restored the thickness of the epidermis damaged by atopic dermatitis. However, no significant differences were observed between the test and control substances. No clinical indicators indicating toxicity due to administration of the test or control substances were observed.

[0258] Meanwhile, the results of visual examination and histopathological examination were as shown in Figs. 19 to 24.

[0259] Through this study, it was confirmed that the test substance AL-BA / BCNF 0.3% cream adhesive transparent wound dressing was non-inferior to the reference substance Estura Atobarrier Itching Cream in terms of safety and efficacy in atopic dermatitis.

[0260] The present invention can be widely used in the fields of health products, pharmaceuticals, medical devices, cosmetics, etc.

Claims

1. A composition for external application to the skin comprising, as an active ingredient, a complex compound of an Al-BA compound in which phenylboronic acid is combined with alginate and a bacterial cellulose nanofiber ((+)BCNF) having a positively charged surface.

2. A skin external application composition according to claim 1, characterized in that the complex compound of the Al-BA compound and (+)BCNF is contained in an amount of 0.1 to 10 wt% based on the total weight of the composition.

3. A skin external application composition according to claim 1, characterized in that the weight ratio of Al-BA complex compound: bacterial cellulose nanofiber (BCNF) = 1:0.01 to 0.

5.

4. A composition for external application to the skin, further comprising one or more moisturizing or skin barrier components selected from glycerin, hyaluronic acid, and cholesterol, according to claim 1.

5. A composition for external application to the skin according to any one of claims 1 to 4, characterized in that the composition is a cosmetic composition for restoring skin barrier function and promoting skin moisturizing.

6. A composition for external application to the skin, characterized in that the composition is a pharmaceutical composition for protecting the skin of a damaged skin barrier area according to any one of claims 1 to 4.

7. A skin external application composition according to claim 6, characterized in that the above-mentioned drug is an adhesive wound dressing medical device.

8. A composition for external application to the skin according to any one of claims 1 to 4, characterized in that the composition is a pharmaceutical composition for treating, preventing, or alleviating one or more diseases or symptoms selected from the group consisting of dry skin, pruritus, dry eczema, hyperkeratosis of the skin, keratosis of the heel and ankle area, xeroderma in the elderly, rough skin in adults, actinic dermatitis, atopic dermatitis, and psoriasis.

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