Composition for external application to skin and pharmaceutical composition, which comprise brussels sprout-derived polydeoxyribonucleotide as active ingredient

Abstract

The present invention relates to a composition for external application to the skin and a pharmaceutical composition, which comprise brussels sprout-derived polydeoxyribonucleotide as an active ingredient.

Description

Topical skin composition and pharmaceutical composition containing polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient

[0001] Cross-citation with related applications

[0002] This application claims the benefit of priority based on Korean Patent Application No. 10-2024-0145762 filed on October 23, 2024, and all contents disclosed in the document of said Korean Patent Application are incorporated herein as part of this specification.

[0003] Technology field

[0004] The present invention relates to a topical skin composition and a pharmaceutical composition containing polydeoxyribonucleotide (PDRN) derived from Brussels sprouts as an active ingredient.

[0005] Human skin undergoes continuous changes as we age. Skin aging is broadly classified into natural aging and extrinsic aging; while natural aging is difficult to control artificially because it is influenced by genetic factors, extrinsic aging is relatively easier to control artificially because it is influenced by environmental factors. Ultraviolet radiation is a representative extrinsic aging factor, and the most prominent phenomenon of extrinsic aging is the formation of wrinkles.

[0006] In vivo, the synthesis and degradation of extracellular matrix materials such as collagen are properly regulated; however, as aging progresses, the synthesis of collagen and elastin decreases. Furthermore, the expression of matrix metalloproteinases (MMPs), which degrade collagen, and elastases, which degrade elastin, is promoted, leading to a decline in skin elasticity and the formation of wrinkles.

[0007] As skin aging progresses, not only do wrinkles increase and elasticity decrease, but the skin's regenerative capacity also declines, pores enlarge or stretch, the skin barrier weakens, skin troubles occur frequently, and melasma and age spots also increase, causing various changes.

[0008] Various studies on anti-aging and slow aging are being conducted to improve and slow down these various changes associated with skin aging.

[0009] Recently, PDRN injections, known as "salmon injections," have been gaining popularity as a type of dermatological procedure. Polydeoxyribonucleotide (PDRN) is typically manufactured using DNA extracted from salmon testes and semen. PDRN induces DNA synthesis and cell regeneration by activating DNA synthesis bypass pathways, and acts as a skin regeneration signaling molecule, adenosine A 2A Receptor (Adenosine A 2A It is known as a component that activates the receptor (ADORA2A) to promote the secretion of various growth factors such as EGF, FGF, and IGF, vasodilation and production of VEGF (vascular endothelial growth factor), and anti-inflammatory effects.

[0010] PDRN is an animal-derived ingredient primarily extracted from salmon and other sources, and therefore has aspects that make it unsuitable as a cosmetic ingredient, which is preferred over plant-based ingredients.

[0011] <Prior Art Literature>

[0012] Republic of Korea Published Patent Application No. 10-2024-0103455

[0013] The present invention aims to provide a topical skin composition and a pharmaceutical composition that have few side effects and are safe for the human body, while having excellent effects in preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, reducing skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation.

[0014] The present invention, for solving the problems described above, provides a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient.

[0015] The above-mentioned polydeoxyribonucleotide derived from Brussels sprouts may be obtained by extracting a polydeoxyribonucleotide component from Brussels sprouts.

[0016] The content of the above-mentioned Brussels sprout-derived polydeoxyribonucleotide may be 0.01 ppm to 100,000 ppm based on the total amount of the above-mentioned topical skin composition.

[0017] The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is adenosine A 2A It may be activating receptors.

[0018] The above-mentioned polydeoxyribonucleotide derived from Brussels sprouts may perform one or more actions selected from inhibiting the secretion of inflammatory cytokines, increasing anti-inflammatory factors, promoting the secretion of growth factors, and promoting collagen production.

[0019] The above topical skin composition may be for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation.

[0020] The above external skin composition may be a cosmetic composition, a hair composition for scalp improvement, or a combination thereof.

[0021] In addition, the present invention, for solving the problems described above, provides a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation.

[0022] The above pharmaceutical composition may be obtained by extracting a polydeoxyribonucleotide component from a Brussels sprout-derived polydeoxyribonucleotide.

[0023] The content of the Brussels sprout-derived polydeoxyribonucleotide in the above pharmaceutical composition may be 0.01 ppm to 100,000 ppm based on the total amount of the above pharmaceutical composition.

[0024] The Brussels sprout-derived polydeoxyribonucleotide of the above pharmaceutical composition is adenosine A 2A It may be activating receptors.

[0025] The Brussels sprout-derived polydeoxyribonucleotide of the above pharmaceutical composition may have one or more actions selected from inhibiting the secretion of inflammatory cytokines, increasing anti-inflammatory factors, promoting the secretion of growth factors, and promoting collagen production.

[0026] The topical skin composition and pharmaceutical composition according to the present invention contain PDRN (Polydeoxyribonucleotide) derived from Brussels sprouts as an active ingredient, thereby containing adenosine A 2AIt has a mechanism of action that selectively acts on receptors and activates DNA synthesis pathways, and is excellent for inhibiting the secretion of inflammatory cytokines, increasing anti-inflammatory factors, promoting the secretion of growth factors, enhancing wound healing ability through increased collagen production, improving skin regeneration, preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, and improving skin elasticity or wrinkles; furthermore, by utilizing plant-derived ingredients, it may be free from side effects.

[0027] The effects of the present invention are not limited to those mentioned above, and other unmentioned effects will be clearly understood by a person skilled in the art from the description below.

[0028] Figure 1 is a microscopic image showing the degree of recovery of cell scratch wounds when HaCaT cells with induced cell scratch wounds were treated with Brussels sprout-derived PDRN.

[0029] FIGS. 2a to 2c are graphs showing the degree of inhibition of the expression of inflammatory cytokines IL-1α, IL-6, and TNF-α when Brussels sprout-derived PDRN according to the present invention is treated to Raw 264.7 cells.

[0030] Figure 3 is a graph showing the degree of increase in the expression of the anti-inflammatory factor IL-10 when Brussels sprout-derived PDRN according to the present invention is treated to Raw 264.7 cells.

[0031] Figure 4 is a graph showing the expression level of the growth factor FGF when treated in fibroblasts.

[0032] Figure 5 is a graph showing the degree of change in transepidermal water loss (TEWL).

[0033] The advantages and features of the present invention and the methods for achieving them will become clear by referring to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below but can be implemented in various different forms. These embodiments are provided merely to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the present invention, and the present invention is defined only by the scope of the claims.

[0034] The terms used in this specification are for describing the embodiments and are not intended to limit the invention. In this specification, the singular form includes the plural form unless specifically stated otherwise in the text. The terms "comprises" and / or "comprising" as used in this specification do not exclude the presence or addition of one or more other components in addition to the components mentioned. Throughout the specification, "and / or" includes each of the mentioned components and all combinations of one or more. Although terms such as "first," "second," etc., are used to describe various components, these components are not limited by these terms. These terms are used merely to distinguish one component from another. Therefore, the first component mentioned below may be the second component within the technical scope of the invention.

[0035] Unless otherwise defined, all terms used herein (including technical and scientific terms) may be used in a meaning commonly understood by those skilled in the art to which the present invention pertains. Additionally, terms defined in commonly used dictionaries are not to be interpreted ideally or excessively unless explicitly and specifically defined otherwise.

[0036]

[0037] The present invention will be described in detail below.

[0038] The present invention provides a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient.

[0039] In the case of a topical skin composition composed of the above ingredients, adenosine A 2A It has a mechanism of action that selectively acts on receptors and activates DNA synthesis pathways, and exhibits excellent effects such as inhibiting the secretion of inflammatory cytokines, increasing anti-inflammatory factors, promoting the secretion of growth factors, increasing collagen production, preventing skin aging through wound healing, soothing the skin, strengthening the skin barrier, improving and shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, skin regeneration, wound healing, and reducing or treating inflammation; furthermore, by utilizing plant-derived ingredients, it may be free from side effects.

[0040] In one embodiment, the Brussels sprout-derived polydeoxyribonucleotide can be obtained by extracting Brussels sprouts.

[0041] The above-mentioned Brussels sprouts are also known as mini cabbages or mini cabbages; their scientific name is Brassica oleracea var. gemmifera, and they belong to the genus Brassica in the family Brassicaceae. The stems grow to a height of 60 to 90 cm, and the cabbage heads form in a bell-like shape from the axillary buds, with a diameter of 3 to 5 cm. Each Brussels sprout weighs 10 to 15 g. The main domestic production areas for Brussels sprouts are southern regions, such as Jeju Island.

[0042] The above-mentioned Brussels sprouts are rich in vitamins A, C, B1, B3, B9, and K, and it is known that glucosinolate components interact with enzymes to produce isothiocyanate, which has anti-inflammatory effects and helps with vascular health. In addition, the above-mentioned Brussels sprouts are also known to have the effect of helping with blood sugar control and digestive function.

[0043] The term "polydeoxyribonucleotide" in the present invention may also be described as PDRN and is a mixture of short deoxyribonucleotides. That is, it is a low molecular weight DNA complex produced by fractionating DNA chains into specific sizes.

[0044] The method for extracting PDRN components from the above-mentioned Brussels sprouts is not particularly limited and can be extracted according to methods commonly used in the relevant technical field.

[0045] The method for extracting PDRN components from the above-described Brussels sprouts may include, for example, the step of washing and grinding the Brussels sprout raw material; the step of dissolving the washed and ground Brussels sprout raw material in a buffer solution; and the step of obtaining PDRN from the buffer. Here, the buffer may include a cetrimonium bromide (CTAB) buffer containing polyvinylpyrrolidone (PVP) and 2-mercaptoethanol. Additionally, in the step of obtaining PDRN from the buffer, high-purity Mugwort-derived PDRN may be obtained by using a spin column or a centrifugation method. Here, using a spin column may be preferable, while a centrifugation method may be used for cost-effectiveness or to obtain a large volume.

[0046] The content of the above-mentioned Brussels sprout-derived polydeoxyribonucleotide may be 0.01 ppm to 100,000 ppm, 0.01 ppm to 10,000 ppm, 0.01 ppm to 1,000 ppm, 0.01 ppm to 100 ppm, or 0.01 ppm to 50 ppm based on the total amount of the above-mentioned topical skin composition. When the Brussels sprout-derived polydeoxyribonucleotide is included within the aforementioned range, adenosine A 2A It may be excellent for receptor activation, inhibition of inflammatory cytokine secretion, increase of anti-inflammatory factors, promotion of growth factor secretion, improvement of wound healing ability due to increased collagen production, enhancement of skin regeneration, anti-aging, skin soothing, strengthening of the skin barrier, improvement of skin pores, and improvement of skin elasticity or wrinkles.

[0047] In one embodiment, the Brussels sprout-derived polydeoxyribonucleotide is adenosine A 2AIt can activate receptors. Known subtypes of the adenosine receptor mentioned above include A1 and A 2A , A 2B , and A3 receptors exist. Among these, adenosine A 2A The receptor is a skin regeneration signaling molecule and is generally known to inhibit inflammatory responses. The above adenosine A 2A It can exhibit various physiological effects, such as promoting the secretion of various growth factors including EGF, FGF, and IGF, which increase gene expression of receptors, vasodilation and production by VEGF (vascular endothelial growth factor), and anti-inflammatory action.

[0048] The above-mentioned polydeoxyribonucleotide derived from Brussels sprouts can inhibit the secretion of inflammatory cytokines or increase anti-inflammatory factors. Examples of such inflammatory cytokines include Interleukin-1α (IL-1α), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-12 (IL-12), Tumor Necrosis Factor-α (TNF-α), and Transforming Growth Factor-β (TGF-β). A topical skin composition containing the Brussels sprouts-derived polydeoxyribonucleotide according to the present invention as an active ingredient can inhibit the expression of inflammatory cytokines IL-1α, IL-6, and TNF-α. Furthermore, the above-mentioned anti-inflammatory factor may be Interleukin-10 (IL-10), which is a cytokine capable of mediating many actions or effects. The above-mentioned anti-inflammatory agent IL-10 can inhibit the secretion of the above-mentioned inflammatory cytokines. Through this, anti-inflammatory and skin soothing effects can be obtained.

[0049] The aforementioned polydeoxyribonucleotide derived from Brussels sprouts can promote the secretion of growth factors. Specifically, the growth factor may be Fibroblast Growth Factor (FGF). Fibroblast Growth Factor (FGF) is an important family of proteins that regulates cell growth, differentiation, survival, migration, tissue development, and angiogenesis. FGF is expressed in various cell types and has a significant effect, particularly on fibroblasts, vascular endothelial cells, neurons, and bone cells.

[0050] The above-mentioned Brussels sprout-derived polydeoxyribonucleotide may promote collagen production. By containing the above-mentioned Brussels sprout-derived polydeoxyribonucleotide that promotes collagen production as an active ingredient, the skin can be made more elastic by suppressing or inhibiting skin sagging caused by internal or external factors such as aging and stress, and, for example, may have effects such as wrinkle improvement and skin elasticity improvement.

[0051] In one embodiment, the topical skin composition may contain a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient and may be for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation.

[0052] For the above-mentioned anti-aging, skin soothing, skin barrier strengthening, skin pore improvement, skin pore reduction, skin elasticity enhancement, skin wrinkle improvement, skin regeneration, wound healing, inflammation improvement, or inflammation treatment, in addition to Brussels sprout-derived PDRN, moisturizers, thickeners, surfactants, oil phase bases, preservatives, antioxidants, alcohols, fragrances, pH adjusters, natural extracts, etc., may be selected and combined without difficulty by a person skilled in the art within a range that does not impair the effects of the present invention, and in this case, a synergistic effect may occur when applied simultaneously with other raw materials.

[0053] In one embodiment, the topical skin composition may be a cosmetic composition, a hair composition for improving the scalp, or a combination thereof.

[0054] The above cosmetic composition may be for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation. The formulation of the above cosmetic composition is not particularly limited and can be appropriately selected according to the purpose. For example, it may be manufactured in the form of a softening lotion, an astringent lotion, a nourishing lotion, a massage cream, an essence, a nourishing cream, a pack, a powder, a wax cream, etc., but is not limited thereto.

[0055] The above hair composition for scalp improvement may be for soothing the scalp and improving the scalp barrier, and may be in the form of a shampoo, rinse, treatment, hair pack, etc., but is not limited thereto.

[0056]

[0057] In addition, the present invention provides a pharmaceutical composition for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation, containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient.

[0058] In one embodiment, the Brussels sprout-derived polydeoxyribonucleotide can be obtained by extracting Brussels sprouts.

[0059] The above-mentioned Brussels sprouts are also known as mini cabbages or mini cabbages; their scientific name is Brassica oleracea var. gemmifera, and they belong to the genus Brassica in the family Brassicaceae. The stems grow to a height of 60 to 90 cm, and the cabbage heads form in a bell-like shape from the axillary buds, with a diameter of 3 to 5 cm. Each Brussels sprout weighs 10 to 15 g. The main domestic production areas for Brussels sprouts are southern regions, such as Jeju Island.

[0060] The above-mentioned Brussels sprouts are rich in vitamins A, C, B1, B3, B9, and K, and it is known that glucosinolate components interact with enzymes to produce isothiocyanate, which has anti-inflammatory effects and helps with vascular health. In addition, the above-mentioned Brussels sprouts are also known to have the effect of helping with blood sugar control and digestive function.

[0061] The method for extracting PDRN components from the above-mentioned Brussels sprouts is not particularly limited and can be extracted according to methods commonly used in the relevant technical field.

[0062] The method for extracting PDRN components from the above-mentioned Brussels sprouts may include, for example, the step of washing and grinding the Brussels sprout raw material; the step of dissolving the washed and ground Brussels sprout raw material in a buffer solution; and the step of obtaining PDRN from the buffer. Here, the buffer may be a cetrimonium bromide (CTAB) buffer containing polyvinylpyrrolidone (PVP) and 2-mercaptoethanol. Additionally, in the step of obtaining PDRN from the buffer, a spin column or a centrifugation method may be used.

[0063] The content of the above-mentioned Brussels sprout-derived polydeoxyribonucleotide may be 0.01 ppm to 100,000 ppm, 0.01 ppm to 10,000 ppm, 0.01 ppm to 1,000 ppm, 0.01 ppm to 100 ppm, or 0.01 ppm to 50 ppm based on the total amount of the above-mentioned pharmaceutical composition. When the Brussels sprout-derived polydeoxyribonucleotide is included within the aforementioned range, adenosine A 2A It may be excellent for receptor activation, inhibition of inflammatory cytokine secretion, increase of anti-inflammatory factors, promotion of growth factor secretion, improvement of wound healing ability due to increased collagen production, enhancement of skin regeneration, anti-aging, skin soothing, strengthening of the skin barrier, improvement of skin pores, and improvement of skin elasticity or wrinkles.

[0064] In one embodiment, the Brussels sprout-derived polydeoxyribonucleotide is adenosine A 2A It can activate receptors. Known subtypes of the adenosine receptor mentioned above include A1 and A 2A , A 2B , and A3 receptors exist. Among these, adenosine A 2AThe receptor is a skin regeneration signaling molecule and is generally known to inhibit inflammatory responses. The above adenosine A 2A It can exhibit various physiological effects, such as promoting the secretion of various growth factors including EGF, FGF, and IGF, which increase gene expression of receptors, vasodilation and production by VEGF (vascular endothelial growth factor), and anti-inflammatory action.

[0065] The above-mentioned polydeoxyribonucleotide derived from Brussels sprouts can inhibit the secretion of inflammatory cytokines or increase anti-inflammatory factors. Examples of such inflammatory cytokines include Interleukin-1α (IL-1α), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-12 (IL-12), Tumor Necrosis Factor-α (TNF-α), and Transforming Growth Factor-β (TGF-β). A pharmaceutical composition containing the Brussels sprouts-derived polydeoxyribonucleotide according to the present invention as an active ingredient can inhibit the expression of inflammatory cytokines IL-1α, IL-6, and TNF-α. Furthermore, the above-mentioned anti-inflammatory factor may be Interleukin-10 (IL-10), which is a cytokine capable of mediating many actions or effects. The above-mentioned anti-inflammatory agent IL-10 can inhibit the secretion of the above-mentioned inflammatory cytokines. Through this, anti-inflammatory and skin soothing effects can be obtained.

[0066] The aforementioned polydeoxyribonucleotide derived from Brussels sprouts can promote the secretion of growth factors. Specifically, the growth factor may be Fibroblast Growth Factor (FGF). Fibroblast Growth Factor (FGF) is an important family of proteins that regulates cell growth, differentiation, survival, migration, tissue development, and angiogenesis. FGF is expressed in various cell types and has a significant effect, particularly on fibroblasts, vascular endothelial cells, neurons, and bone cells.

[0067] The above-mentioned Brussels sprout-derived polydeoxyribonucleotide may promote collagen production. By containing the above-mentioned Brussels sprout-derived polydeoxyribonucleotide that promotes collagen production as an active ingredient, the skin can be made more elastic by suppressing or inhibiting skin sagging caused by internal or external factors such as aging and stress, and, for example, may have effects such as wrinkle improvement and skin elasticity improvement.

[0068] The above pharmaceutical composition may be a transdermal formulation such as a lotion, ointment, gel, cream, patch, etc., but is not limited thereto.

[0069] The above pharmaceutical composition may be for improving the skin barrier and for skin regeneration. The above pharmaceutical composition may be used for skin barrier and skin regeneration-related diseases, for example, atopic dermatitis, dermatitis caused by rough skin, heat rash, blistering, frostbite, diaper-induced skin disease, contact dermatitis, seborrheic dermatitis, Vidal lichen, nummular eczema, housewife's eczema, photodermatitis, cystic dermatitis, skin atrophy, prurigo, drug rash, toxic rash, psoriasis, psoriasis psoriasis, palmar pustulosis, lichen planus, glossy lichen, follicular erythematous

[0070] The topical skin composition and pharmaceutical composition according to one embodiment may further include, in addition to the above components, conventional additives used in the art within a range that does not impede the purpose of the present invention, and the types and amounts thereof may be appropriately selected and adjusted by a person skilled in the art.

[0071]

[0072] The present invention provides a use for a polydeoxyribonucleotide derived from Brussels sprouts as a topical skin preparation composition.

[0073] The present invention relates to an adenosine A of a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient. 2A It provides a use for activating receptors.

[0074] The present invention provides a use for a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient for inhibiting the secretion of inflammatory cytokines.

[0075] The present invention provides a use for increasing anti-inflammatory factors in a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient.

[0076] The present invention provides a use for a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient for promoting the secretion of growth factors.

[0077] The present invention provides a use for a topical skin composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient for promoting collagen production.

[0078] The present invention provides a use for a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation.

[0079]

[0080] The present invention relates to a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient, applied to the skin of an animal to produce adenosine A 2A It provides a method to activate receptors.

[0081] The present invention provides a method for inhibiting the secretion of inflammatory cytokines by applying a topical skin composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0082] The present invention provides a method for increasing anti-inflammatory factors by applying a topical skin composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0083] The present invention provides a method for promoting the secretion of growth factors by applying a topical skin composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0084] The present invention provides a method for promoting collagen production by applying a topical skin composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0085] The present invention provides a method for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation by applying a topical skin composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0086] The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is as described above.

[0087] The above animal could be a human.

[0088]

[0089] The present invention provides a method for preparing a topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient.

[0090] The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is as described above.

[0091]

[0092] The present invention provides a use for a polydeoxyribonucleotide derived from Brussels sprouts in a pharmaceutical composition.

[0093] The present invention relates to adenosine A of a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient. 2A It provides a use for activating receptors.

[0094] The present invention provides a use for a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient for inhibiting the secretion of inflammatory cytokines.

[0095] The present invention provides a use for increasing anti-inflammatory factors in a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient.

[0096] The present invention provides a use for a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient for promoting the secretion of growth factors.

[0097] The present invention provides a use for a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient for promoting collagen production.

[0098] The present invention provides a use for a pharmaceutical composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation.

[0099]

[0100] The present invention relates to a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient, applied to the skin of an animal to produce adenosine A 2A It provides a method to activate receptors.

[0101] The present invention provides a method for inhibiting the secretion of inflammatory cytokines by applying a pharmaceutical composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0102] The present invention provides a method for increasing anti-inflammatory factors by applying a pharmaceutical composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0103] The present invention provides a method for promoting the secretion of growth factors by applying a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient to the skin of an animal.

[0104] The present invention provides a method for promoting collagen production by applying a pharmaceutical composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0105] The present invention provides a method for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation by applying a pharmaceutical composition containing a Brussels sprout-derived polydeoxyribonucleotide as an active ingredient to the skin of an animal.

[0106] The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is as described above.

[0107] The above animal could be a human.

[0108]

[0109] The present invention provides a method for preparing a pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient.

[0110] The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is as described above.

[0111]

[0112] Specific embodiments of the present invention are presented below. However, the embodiments described below are merely for the purpose of specifically illustrating or explaining the present invention and should not be limited thereto. Furthermore, details not described herein can be sufficiently technically inferred by a person skilled in the art, so their description is omitted.

[0113] [Preparation Example]

[0114] Preparation Example 1: Preparation of PDRN derived from Brussels sprouts

[0115] After washing the Brussels sprouts, they were crushed using a mortar and pestle with liquid nitrogen. To 50 mg of finely crushed Brussels sprouts, 390 µl of Lysis buffer (EDTA, SDS, NaCl, Tris base, Tween 20), 20 µl of Proteinase K solution, and 5 µl of RNase A solution were added and mixed well, then reacted at 65°C for 30 minutes. An additional 100 µl of Precipitation Solution was added and mixed well, then reacted on ice for 5 minutes. After centrifugation at 13,000 rpm for 5 minutes at room temperature, 200 µl of the supernatant was taken and transferred to a new tube. An additional 650 µl of Binding Solution (Ethanol, EDTA, Tris Base) was added and mixed well, then 650 µl was taken and carefully transferred to the top of the spin column. The solution obtained after centrifugation at 13,000 rpm for 1 minute at room temperature was discarded, and 200 µl of the remaining solution was filtered in the same manner. 700 µl of primary washing buffer was added to the top of the spin column, and washing was performed by centrifuging at 13,000 rpm for 1 minute. After adding 700 µl of secondary washing buffer to the top of the spin column and performing a second wash by centrifuging at 13,000 rpm for 1 minute, the column was dried by centrifuging an additionally at 13,000 rpm for 1 minute. A new tube was placed beneath the spin column, and 100 µl of elution buffer (Tris Base) was carefully dispensed onto the column membrane, followed by an incubation at room temperature for 1 minute. Afterwards, the PDRN solution of Brussels sprouts that passed through the tube was obtained by centrifuging at 13,000 rpm for 1 minute, and the obtained PDRN solution was further treated with ultrasound to obtain PDRN of Brussels sprouts at a low molecular level.The concentration of PDRN was confirmed by quantifying it using Nanodrop, and PDRN was verified through agarose gel electrophoresis. The obtained Brussels sprout-derived PDRN has a size of 1500 bp or less and a molecular weight of 55 kDa or less.

[0116]

[0117] [Examples and Comparative Examples]

[0118] Examples 1 to 2 and Comparative Examples 1 to 2: Compositions comprising Brussels sprout-derived PDRN

[0119] A cream formulation composition was prepared by mixing each component according to the compositions in Tables 1 and 2 below, wherein the total amount of each composition is 100% by weight.

[0120] Ingredient Name Content (Weight%) Comparative Example 1 Example 1 PDRN derived from Brussels sprouts - 0.5 ppm Glycerin 20 20 Hyaluronic acid 55 Olive oil 55 Cetyl ethyl palmitate 55 PEG-100 stearate, Glyceryl stearate 22 Polysorbate 60 11 Sorbitan sesquioleate 0.5 0.5 Trehalose 22 1,2-Hexanediol 22 Dimethicone 11 Carbomer 0.15 0.15 Tromethamine 0.1 0.1 EDTA-2Na 0.0 2 0.0 2 Purified water Remaining amount Remaining amount

[0121] Ingredient Name Content (Weight%) Comparative Example 2 Example 2 Brussels Sprout-derived PDRN-1 ppm Cholesterol 0.5 0.5 Stearic Acid 1.5 1.5 Cetearyl Alcohol 2.0 2.0 Polyglyceryl-10 Distearate 2.0 2.0 Caprylyl / Capryl Triglyceride 7.0 7.0 Thickener 0.1 0.1 Purified Water Remaining Amount Remaining Amount

[0122]

[0123] [Experimental Example]

[0124] Experimental Example 1: Confirmation of wound healing and skin regeneration effects

[0125] Human HaCaT keratinocytes were inoculated into DMEM medium containing 10% (v / v) FBS, 100 U / mL penicillin, and 100 µg / mL streptomycin, and cultured at 37°C under 5% CO2. While the in vitro wound healing assay typically utilizes a pipette-tip-assisted cell scratching method, cell scratching was induced using an ibidi culture-insert 2-well to address the issue of reproducibility regarding cell scratch intervals. After detaching HaCaT cells by trypsin treatment, 3 to 7 x 10⁶ 5 70 μL was injected into each culture-insert well to a cell / mL concentration. After incubating for 24 hours at 37°C under 5% CO2, the culture-insert was removed and the cells were washed with PBS. To exclude the effect of cell growth, 5 μg / mL of mitomycin C was treated for 1 hour and then washed with PBS.

[0126] 0.01 ppm of Brussels sprout-derived PDRN prepared in Preparation Example 1 above was diluted in DMEM medium and treated with the above-mentioned HaCaT cells with induced cell scratch wounds. The negative control group used culture medium not treated with Brussels sprout-derived PDRN. The degree of recovery of the cell scratch wounds was observed over time using a 4x magnification microscope, and the results are shown in Figure 1 below. As can be seen from the results in Figure 1, it was confirmed that after 22 hours, the scratched area of ​​the cells treated with Brussels sprout-derived PDRN was recovered more significantly in the cells compared to the negative control group. Therefore, it can be seen that the Brussels sprout-derived PDRN of the present invention has excellent skin wound healing and skin regeneration effects. The wound area was quantified using the Image J program, and the wound area reduction rate was calculated using Equation 1, with the results shown in Table 3.

[0127] [Mathematical Formula 1]

[0128] Wound Area Reduction Rate (%) = [(A0-A T ) / A0] X 100

[0129] A0: Initial wound area

[0130] A T : Wound area after a certain period of time

[0131] Wound area reduction rate (%) Negative control 21 Brussels sprout-derived PDRN 0.01 ppm 41

[0132] Referring to Table 3 above, it can be seen that the Brussels sprout-derived PDRN according to the present invention has excellent skin wound healing and skin regeneration effects.

[0133]

[0134] Experimental Example 2: Confirmation of Increased Collagen Synthesis

[0135] The effect of increasing intracellular collagen production was measured using Human Dermal Fibroblasts (HDFs). HDFs cells were cultured in Medium 106 supplemented with 0.1% P / S and LGSG (Low Serum Growth Supplement) in a 37°C, 5% CO2 incubator. HDFs cells were placed in 2×10⁻¹⁰ 12-well plates. 4Cells were inoculated into wells and cultured for 24 hours. Subsequently, the medium was replaced with one containing Brussels sprout-derived PDRN prepared in Preparation Example 1 at concentrations of 0.01, 0.05, 0.1, and 5 ppm, and cultured for another 24 hours. TGF-β1 (0.0004 μM) was used as the positive control, and the reaction solution without added samples was used as the negative control. After 24 hours of culture, the supernatant was collected, and the amount of procollagen released in the medium was measured by absorbance at 450 nm using a Procollagen Type I C-Peptide (PIP) ELISA Kit. The degree of collagen production was evaluated by adjusting for total protein content. The experimental results regarding the effect of increasing intracellular collagen production in the negative control group are shown in Table 4.

[0136] Collagen production increase rate (%) Negative control 0 Brussels sprout-derived PDRN 0.01 ppm 30 Brussels sprout-derived PDRN 0.05 ppm 42 Brussels sprout-derived PDRN 0.1 ppm 49 Brussels sprout-derived PDRN 1 ppm 73 Brussels sprout-derived PDRN 5 ppm 82 TGF-β152

[0137] Referring to Table 4 above, the collagen-producing effect of the Brussels sprout-derived PDRN according to the present invention showed excellent results when compared to the positive control group. Based on this, it can be expected that the composition of the present invention is effective in improving skin wrinkles and skin elasticity by effectively promoting collagen.

[0138]

[0139] Experimental Example 3: Adenosine A 2A Confirmation of receptor activation

[0140] Adenosine A produced by Brussels sprout-derived PDRN prepared in Preparation Example 1 above 2A Receptor (Adenosine A 2ATo evaluate the activation of the receptor, human epidermal keratinocytes (HEK) were first cultured in EpiLife medium containing 0.1% P / S and HKGS (Human Keratinocyte Growth Supplement) in a 37°C, 5% CO2 incubator. HEK cells were cultured at a rate of 1 x 10⁻¹⁰ 5 After inoculating a 6-well plate at a cell / well concentration, 24 hours later, human keratinocytes were treated with Brussels sprout-derived PDRN at concentrations of 0.01, 0.1, 1, and 50 ppm.

[0141] To measure gene expression levels, total RNA was extracted from cells using the easy-spin™ Total RNA extraction kit (iNtRoN, Korea). cDNA was synthesized from the extracted RNA using Maxime RT PreMix (iNtRON, Korea), and Real-time PCR was performed using the QuantStudio 6 Flex Real-Time PCR system (Applied Biosystems, USA) to analyze adenosine A 2A The gene expression level of the receptor was checked. At this time, a group not treated with the Brussels sprout-derived PDRN prepared in Preparation Example 1 was used as the negative control, and mRNA of β-actin was used as the internal control.

[0142] Gene expression levels were expressed as a ratio compared to the control group value (control-treated = 1), and the results are shown in Table 5 below.

[0143] (control-treated = 1) Adenosine A 2A Receptor Brussels sprout-derived PDRN 0.01 ppm 2.1 Brussels sprout-derived PDRN 0.1 ppm 2.9 Brussels sprout-derived PDRN 1 ppm 3.6 Brussels sprout-derived PDRN 50 ppm 5.0

[0144] Referring to Table 5 above, adenosine A by treatment with Brussels sprout-derived PDRN according to the present invention 2A It was confirmed that the gene expression of the receptor increased. Therefore, Brussels sprout-derived PDRN adenosine A 2A It can be seen that various physiological effects are exhibited through receptors.

[0145]

[0146] Experimental Example 4: Confirmation of Anti-inflammatory Effect

[0147] To evaluate the inhibition of inflammatory cytokine expression by Brussels sprout-derived PDRN according to Preparation Example 1 above, murine macrophage cell line Raw 264.7 cells were first cultured in DMEM medium containing 1% P / S and 5% FBS at 37°C in a 5% CO2 incubator. 5X10 4 After inoculating 6-well plates at a cell / well concentration, 24 hours later, lipopolysaccharide (LPS) 1 μg / mL and Brussels sprout-derived PDRN prepared in Preparation Example 1 were applied to Raw 264.7 cells at concentrations of 0.01, 0.1, 1, and 10 ppm, respectively. After 48 hours of incubation, the supernatant was collected, and the expression levels of IL-1α, IL-6, IL-10, and TNF-α were measured using an Interleukin-1α ELISA kit (R&D systems, USA), an Interleukin-6 ELISA kit (R&D systems, USA), an Interleukin-10 ELISA kit (R&D systems, USA), and a TNF-α ELISA kit (Abcam, USA). At this time, a group not treated with either LPS or Brussels sprout-derived PDRN prepared in Preparation Example 1 was used as the negative control.

[0148] The results are shown in Figs. 2a to 2c and Fig. 3. Figs. 2a, 2b, and 2c are graphs showing the degree of inhibition of the expression of inflammatory cytokines IL-1α, IL-6, and TNF-α, respectively, when Brussels sprout-derived PDRN according to the present invention is treated to Raw 264.7 cells. Fig. 3 is a graph showing the degree of increase in the expression of the anti-inflammatory factor IL-10 when Brussels sprout-derived PDRN according to the present invention is treated to Raw 264.7 cells. Referring to Figs. 2a to 2c and Fig. 3, it was confirmed that the expression of inflammatory cytokines IL-1α, IL-6, and TNF-α is inhibited by Brussels sprout-derived PDRN according to the present invention, and the expression of the anti-inflammatory factor IL-10 is increased.

[0149] Through this, the composition of the present invention can be expected to have anti-inflammatory and skin-soothing effects.

[0150]

[0151] Experimental Example 5: Confirmation of Increased Growth Factor Expression

[0152] The effect of increasing the expression of the growth factor FGF (Fibroblast Growth Factor) was measured using Human Dermal Fibroblasts (HDFs). HDFs cells were cultured in Medium 106 supplemented with 0.1% P / S and LGSG (Low Serum Growth Supplement) in a 37°C, 5% CO2 incubator. HDFs cells were placed in a 6-well plate at 5×10 4 After inoculating cells / well and culturing for 24 hours, the medium was replaced with one containing Brussels sprout-derived PDRN prepared in Preparation Example 1 at concentrations of 0.01, 0.05, 0.1, and 5 ppm, and cultured for 48 hours. A reaction solution without added sample was used as the negative control.

[0153] After culturing for 48 hours, the supernatant was taken and the amount of free FGF in the medium was measured using an FGF ELISA Kit, and the experimental results of the effect of increasing intracellular FGF production compared to the negative control group are shown in Figure 4.

[0154] Referring to Fig. 4, it was confirmed that when fibroblasts were treated with PDRN derived from Brussels sprouts according to the present invention, the amount of growth factor FGF produced increased compared to the negative control group.

[0155]

[0156] Experimental Example 6: Safety test for skin

[0157] A human patch test was performed to evaluate the degree of skin irritation. The patch test was conducted on 30 healthy individuals aged 20 years or older. Samples were prepared as 10,000 and 100,000 ppm aqueous solutions and applied to the inner upper arm using a Finn chamber for 48 hours. After removal, the first reading was taken, and a second reading was performed after 72 hours to determine the skin reaction results. The evaluation criteria are as shown in Table 6 below, and the skin irritation index was calculated using the following mathematical formula 2. The results are shown in Table 7.

[0158] Reaction Weighting Criteria -0.0 No reaction ±0.5 Suspected positive reaction +1.0 Weak positive reaction ++2.0 Strong positive reaction +++3.0 Extremely positive reaction

[0159] [Mathematical Formula 2] Skin Irritation Index = Σ(Weight X Number of Response Factors) / (Maximum Weight X Total Number of Subjects) × 100

[0160] Skin Irritation Index (Irritation Level) Brussel Sprout-derived PDRN (10,000 ppm) 0.0 Non-irritating Brussel Sprout-derived PDRN (100,000 ppm) 0.0 Non-irritating

[0161] Referring to Table 7 above, the Brussels sprout-derived PDRN according to the present invention was found to be non-irritating to the skin at 100,000 ppm or less, and it was confirmed that there is no irritation to the skin when used as an external skin preparation.

[0162]

[0163] Experimental Example 7: Measurement of skin elasticity, wrinkle improvement, and pore reduction effects

[0164] Skin elasticity, wrinkle improvement, and pore reduction effects were measured for the compositions prepared in Example 1 and Comparative Example 1 above.

[0165] Specifically, to investigate the skin elasticity enhancement, skin wrinkle improvement, and pore reduction effects of a composition containing Brussels sprout-derived PDRN as an active ingredient according to the present invention, a human application test was conducted on 20 women. The compositions prepared in Comparative Example 1 and Example 1 were divided into two groups of 10 people each and used twice a day, once in the morning and once in the evening, for 4 weeks.

[0166] Then, to measure the skin elasticity enhancement effect after using the product, the elasticity of the facial area was measured using a Cutometer (Cutometer, Courage+Khazaka Electronic GmbH., Cologne, Germany), a device that measures skin elasticity by applying and releasing negative pressure to the skin, and compared with the result before application, and the results are shown in Table 8 below.

[0167] Skin elasticity improvement (%) Comparative Example 124.9 Example 143.8

[0168] The degree of improvement in skin wrinkles before and after use was measured using the Facial Stage DM-3 (Moritex, Japan) pre-face imaging system, and the effect was compared by calculating the change in wrinkle area after 4 weeks using the following Equation 3 with an analysis program. The results are shown in Table 9 below.

[0169] [Mathematical Formula 3]

[0170] Change in wrinkle area = Wrinkle area before use - Wrinkle area after 4 weeks of use

[0171] Change in wrinkle area (mm) 2 )Comparative Example 12.5 Example 120.6

[0172] The skin pore-shrinking effect before and after use was measured using the Facial Stage DM-3 (Moritex, Japan) pre-face imaging system, and the effect was compared by calculating the change in pore area after 4 weeks using the following Equation 4 with an analysis program. The results are shown in Table 10 below.

[0173] [Mathematical Formula 4]

[0174] Change in pore area = Pore area before use - Pore area after 4 weeks of use

[0175] Change in pore area (mm) 2 )Comparative Example 12.5 Example 125.2

[0176] Referring to Tables 8 to 10 above, it can be seen that when the composition according to Example 1 containing Brussels sprout-derived PDRN according to the present invention is used for 4 weeks, it has excellent effects in improving skin elasticity, wrinkle improvement, and skin pore improvement compared to Comparative Example 1.

[0177]

[0178] Experimental Example 8: Skin barrier improvement test through measurement of transepidermal water loss (TEWL)

[0179] To confirm the skin barrier improvement effect of the composition according to the present invention, the composition according to Comparative Example 2 and Example 2 was applied to the skin and evaluated by measuring transepidermal water loss (TEWL).

[0180] Specifically, 10 adults aged 30 to 43 years were tested on two areas of 1.5 cm x 1.5 cm on the inner side of the forearm, using the compositions of Example 2 and Comparative Example 2 as test sites.

[0181] First, the transepidermal water loss of the test site was measured using a transepidermal water loss measuring device (Vapometer, Delfin Technologies, Finland), and then the skin barrier was artificially damaged by removing the stratum corneum of the test site through tape stripping. After the skin barrier was damaged, the compositions of Example 2 and Comparative Example 2 were applied to the test site twice a day, and the transepidermal water loss was measured after 1, 3, and 5 days, and the results are shown in Fig. 5.

[0182] Figure 5 is a graph showing the degree of change in transepidermal water loss when the composition according to Example 2 and Comparative Example 2 is applied.

[0183] Referring to FIG. 5, when the composition of Example 2 containing Brussels sprout-derived PDRN according to one embodiment was applied, the transepidermal water loss value was reduced compared to Comparative Example 2, indicating excellent skin barrier improvement effect.

[0184]

[0185] Although embodiments of the present invention have been described above, those skilled in the art will understand that the present invention may be implemented in other specific forms without altering its technical concept or essential features. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. A topical skin composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient.

2. In Paragraph 1, The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is a topical skin composition obtained by extracting a polydeoxyribonucleotide component from Brussels sprouts.

3. In Paragraph 1, A topical skin composition in which the content of the above-mentioned Brussels sprout-derived polydeoxyribonucleotide is 0.01 ppm to 100,000 ppm based on the total amount of the above-mentioned topical skin composition.

4. In Paragraph 1, The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is adenosine A 2A A topical skin composition that activates receptors.

5. In Paragraph 1, The above-mentioned polydeoxyribonucleotide derived from Brussels sprouts is a topical skin composition having one or more actions selected from inhibiting the secretion of inflammatory cytokines, increasing anti-inflammatory factors, promoting the secretion of growth factors, and promoting collagen production.

6. In Paragraph 1, The above-mentioned topical skin composition is a topical skin composition for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation.

7. In Paragraph 1, The above-mentioned topical skin composition is a cosmetic composition, a hair composition for scalp improvement, or a combination thereof.

8. A pharmaceutical composition containing a polydeoxyribonucleotide derived from Brussels sprouts as an active ingredient for preventing skin aging, soothing the skin, strengthening the skin barrier, improving skin pores, shrinking skin pores, enhancing skin elasticity, improving skin wrinkles, regenerating the skin, healing wounds, improving inflammation, or treating inflammation.

9. In Paragraph 8, The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is a pharmaceutical composition obtained by extracting a polydeoxyribonucleotide component from Brussels sprouts.

10. In Paragraph 8, A pharmaceutical composition in which the content of the above-mentioned Brussels sprout-derived polydeoxyribonucleotide is 0.01 ppm to 100,000 ppm based on the total amount of the above-mentioned pharmaceutical composition.

11. In Paragraph 8, The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is adenosine A 2A A pharmaceutical composition that activates receptors.

12. In Paragraph 8, The above-mentioned Brussels sprout-derived polydeoxyribonucleotide is a pharmaceutical composition having one or more actions selected from inhibiting the secretion of inflammatory cytokines, increasing anti-inflammatory factors, promoting the secretion of growth factors, and promoting collagen production.

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