Pharmaceutical composition containing pegylated bilirubin for preventing or treating inflammatory diseases

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating inflammatory diseases, which comprises a compound of Chemical Formula 1, and a cosmetic composition containing the compound. The compound of Chemical Formula 1 can be used to remove reactive oxygen species without toxicity, suppress inflammatory cytokines, protect cells, and reduce inflammation.

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition containing pegylated bilirubin for preventing or treating inflammatory diseases. [Background technology]

[0002] Reactive oxygen species (ROS) or reactive oxygen species (ROS) refer to chemically reactive molecules containing oxygen atoms. ROS are oxygen compounds produced in living organisms, including peroxygen ions and hydrogen peroxide, which contain unpaired electrons. As a result, they are highly reactive and equivalent to oxygen with extremely strong oxidizing power that attacks living tissues and damages cells. Such ROS can also be produced during normal metabolic processes and have been reported to play a role in regulating cellular signaling and homeostasis.

[0003] Oxidative stress refers to damage to normal cells caused by increased concentrations of reactive oxygen species. Oxidative stress reflects an imbalance between the production of reactive oxygen species and the biological ability to detoxify reactive intermediates and repair damage caused by reactive oxygen species. If the imbalance caused by oxidative stress is severe, cell death may occur, and increased reactive oxygen species may directly damage the DNA present in an individual. This can result in various inflammatory responses, aging, hyperlipidemia, chronic fatigue, vascular disease, and other conditions, and may worsen existing diseases. Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to provide a pharmaceutical composition containing pegylated bilirubin for preventing or treating inflammatory diseases.

[0005] An object of the present invention is to provide a cosmetic composition containing PEGylated bilirubin. [Means for solving the problem]

[0006] 1. A pharmaceutical composition for preventing or treating an inflammatory disease, comprising a compound represented by Chemical Formula 1, a solvate thereof, or a pharmaceutically acceptable salt thereof. [ka] (In the above formula 1, R1 and R4 are vinyl groups or methyl groups, and R2 and R3 are methyl groups or vinyl groups, but not all of them need to be vinyl groups or methyl groups.) R5 is polyethylene glycol (PEG) or a derivative thereof.

[0007] 2. A pharmaceutical composition for preventing or treating an inflammatory disease according to item 1 above, wherein the compound is a compound represented by any one of chemical formulas 2 to 7. [ka] [ka] [ka] [ka] [ka] [ka]

[0008] 3. A pharmaceutical composition for preventing or treating an inflammatory disease according to item 1, comprising nanoparticles formed by self-assembly of the compound represented by Chemical Formula 1.

[0009] 4. The pharmaceutical composition for preventing or treating an inflammatory disease according to item 3, wherein the nanoparticles have a size of 1 nm to 5,000 nm.

[0010] 5. In the above item 1, the polyethylene glycol derivative may be methoxy PEG, succinimide of PEG propionic acid, succinimide of PEG butanoic acid, branched PEG-NHS, PEG succinimidyl succinate, succinimide of carboxymethylated PEG, benzotriazole carbonate of PEG, PEG-glycidyl ether, PEG-oxycarbonylimidazole, PEG nitrophenyl carbonates, PEG aldehyde, PEG succinimidyl carboxymethyl ester, and PEG succinimidyl ester. 1. A pharmaceutical composition for preventing or treating an inflammatory disease, comprising a compound selected from the group consisting of:

[0011] 6. A pharmaceutical composition for the prevention or treatment of an inflammatory disease according to item 1, wherein the inflammatory disease is at least one selected from the group consisting of inflammatory skin diseases, osteoarthritis, hepatitis, pneumonia, keratitis, gastritis, nephritis, tuberculosis, bronchitis, pleuritis, peritonitis, spondylitis, pancreatitis, inflammatory bowel disease, urethritis, cystitis, inflammatory arteriosclerosis, sepsis, periodontitis, gingivitis, and autoinflammatory diseases.

[0012] 7. In the above item 6, the inflammatory skin disease is atopic dermatitis, contact dermatitis, psoriasis, seborrheic dermatitis, pruritus, parapsoriasis, urticaria, lichen planus, sunburn, radiodermatitis, erythema multiforme, erythema nodosum, granuloma annulare, keratosis pilaris, xeroderma, panniculitis, pyoderma gangrenosum, acne, A pharmaceutical composition for the prevention or treatment of at least one inflammatory disease selected from the group consisting of rosacea, lupus erythematosus, pemphigus, diaper dermatitis, pityriasis rosea, alopecia areata, androgenic alopecia, vitiligo, and decubitus ulcer.

[0013] 8. A pharmaceutical composition for preventing or treating an inflammatory disease according to item 1, wherein the inflammatory disease is caused by increased oxidative stress.

[0014] 9. A cosmetic composition comprising a compound represented by Chemical Formula 1, a solvate thereof, or a cosmetically acceptable salt thereof. [ka] (In the above formula 1, R1 and R4 are vinyl groups or methyl groups, and R2 and R3 are methyl groups or vinyl groups, but not all of them need to be vinyl groups or methyl groups.) R5 is polyethylene glycol (PEG) or a derivative thereof.

[0015] 10. A cosmetic composition according to item 9, wherein the compound is a compound represented by any one of chemical formulas 2 to 7.

[0016] 11. A cosmetic composition according to item 9, comprising nanoparticles formed by self-assembly of the compound represented by chemical formula 1.

[0017] 12. The cosmetic composition according to item 11, wherein the nanoparticles have a size of 1 nm to 5,000 nm.

[0018] 13. In the above-mentioned item 9, the polyethylene glycol derivative is selected from the group consisting of methoxy PEG, succinimide of PEG propionic acid, succinimide of PEG butanoic acid, branched PEG-NHS, PEG succinimidyl succinate, succinimide of carboxymethylated PEG, benzotriazole carbonate of PEG, PEG-glycidyl ether, PEG-oxycarbonylimidazole, PEG nitrophenyl carbonates, PEG-aldehyde, and PEG succinimidyl carboxymethyl ester. PEG succinimidyl ester, and PEG succinimidyl ester.

[0019] 14. A cosmetic composition according to item 9, wherein the cosmetic composition is for anti-inflammatory or antioxidant purposes.

[0020] 15. A cosmetic composition according to item 9, wherein the cosmetic composition is for preventing or improving skin inflammation caused by increased oxidative stress.

[0021] 16. In the above item 15, the skin inflammation caused by increased oxidative stress is atopic dermatitis, contact dermatitis, psoriasis, seborrheic dermatitis, pruritus, parapsoriasis, urticaria, lichen planus, sunburn, radiation dermatitis, erythema multiforme, erythema nodosum, granuloma annulare, keratosis pilaris, xeroderma, panniculitis, and pyoderma gangrenosum. The cosmetic composition is for treating at least one skin condition selected from the group consisting of acne, rosacea, lupus erythematosus, pemphigus, diaper dermatitis, pityriasis rosea, alopecia areata, androgenic alopecia, vitiligo, and decubitus ulcer.

[0022] 17. A cosmetic composition according to item 9, wherein the cosmetic composition is used for moisturizing the skin, restoring the skin barrier function, or preventing skin aging. [Effects of the Invention]

[0023] By containing PEGylated bilirubin, the composition of the present invention can selectively target inflamed tissues and cells and effectively remove reactive oxygen species that cause inflammation without causing cytotoxicity. [Brief explanation of the drawings]

[0024] [Figure 1] FIG. 1 shows the results of molecular weight analysis (QTOF mass spectrometry) data for the synthesized compounds of chemical formulas 2 to 7. [Figure 2a] FIG. 2a shows the results of dynamic light scattering (DLS) to confirm whether PEGylated bilirubin nanoparticles were formed. [Figure 2b] FIG. 2b shows the results of dynamic light scattering (DLS) to confirm whether PEGylated bilirubin nanoparticles were formed. [Figure 3a] FIG. 3a shows the results of confirming whether or not Dil dye was loaded and whether or not PEGylated bilirubin nanoparticles were formed, using dynamic light scattering (DLS) and changes in absorbance. [Figure 3b] FIG. 3b shows the results of confirming whether or not Dil dye was loaded and whether or not PEGylated bilirubin nanoparticles were formed by dynamic light scattering (DLS) and changes in absorbance. [Figure 3c] FIG. 3c shows the results of confirming whether or not Dil dye was loaded and whether or not PEGylated bilirubin nanoparticles were formed by dynamic light scattering (DLS) and changes in absorbance. [Figure 4a] FIG. 4a shows the results of confirming the increase in intracellular influx of Brixelle in a human epidermal keratinocyte cell line (HaCaT) upon inflammatory conditions (stimulation with house dust mite (HDM) antigen). [Figure 4b] 4b shows the results of confirming the increase in intracellular influx of Brixelle in a human epidermal keratinocyte cell line (HaCaT) under inflammatory conditions (stimulation with house dust mite (HDM) antigen). [Figure 5a]Figure 5a shows the results of treating a human epidermal keratinocyte cell line (HaCaT) with Brixelle under pro-inflammatory conditions (HDM or C48 / 80 stimulation), confirming that Brixelle can remove reactive oxygen species within skin cells and protect them. [Figure 5b] Figure 5b shows the results of Brixelle treatment of a human epidermal keratinocyte cell line (HaCaT) under proinflammatory conditions (HDM or C48 / 80 stimulation), confirming that Brixelle can remove reactive oxygen species within skin cells and protect them. [Figure 5c] Figure 5c shows the results of treating a human epidermal keratinocyte cell line (HaCaT) with Brixelle under pro-inflammatory conditions (HDM or C48 / 80 stimulation), confirming that Brixelle can remove reactive oxygen species within skin cells and protect them. [Figure 5d] Figure 5d shows the results of treating a human epidermal keratinocyte cell line (HaCaT) with Brixelle under pro-inflammatory conditions (HDM or C48 / 80 stimulation), confirming that Brixelle can remove reactive oxygen species within skin cells and protect them. [Figure 6a] Figure 6a shows the results of Brixelle dose-dependently suppressing cytokines (TSLP, IL-25, IL-33) produced in human epidermal keratinocyte cell line (HaCaT) under proinflammatory conditions (HDM or C48 / 80 stimulation). [Figure 6b] Figure 6b shows the results confirming that Brixelle dose-dependently suppresses cytokines (TSLP, IL-25, IL-33) produced in human epidermal keratinocyte cell line (HaCaT) under proinflammatory conditions (HDM or C48 / 80 stimulation). [Figure 6c] Figure 6c shows the results confirming that Brixelle dose-dependently suppresses cytokines (TSLP, IL-25, IL-33) produced in human epidermal keratinocyte cell line (HaCaT) under proinflammatory conditions (HDM or C48 / 80 stimulation). [Figure 6d]Figure 6d shows the results confirming that Brixelle dose-dependently suppresses cytokines (TSLP, IL-25, IL-33) produced in human epidermal keratinocyte cell line (HaCaT) under proinflammatory conditions (HDM or C48 / 80 stimulation). [Figure 6e] Figure 6e shows the results confirming that Brixelle dose-dependently suppresses cytokines (TSLP, IL-25, IL-33) produced in human epidermal keratinocyte cell line (HaCaT) under proinflammatory conditions (HDM or C48 / 80 stimulation). [Figure 6f] Figure 6f shows the results confirming that Brixelle dose-dependently suppresses cytokines (TSLP, IL-25, IL-33) produced in human epidermal keratinocyte cell line (HaCaT) under proinflammatory conditions (HDM or C48 / 80 stimulation). [Figure 7a] Figure 7a shows the results of a study confirming that Brixelle treatment of a reconstructed human epidermis model (KeraSkin™) under inflammatory conditions (UVB irradiation) restored skin barrier function and suppressed inflammatory responses in a dose-dependent manner. [Figure 7ba] Figure 7ba shows the results of confirming that Brixelle treatment of a reconstructed human epidermis model (KeraSkin™) under inflammatory conditions (UVB irradiation) dose-dependently restored skin barrier function and suppressed inflammatory responses. [Figure 7bb] Figure 7bb shows the results of confirming that Brixelle treatment of a reconstructed human epidermis model (KeraSkin™) under inflammatory conditions (UVB irradiation) dose-dependently restored skin barrier function and suppressed inflammatory responses. [Figure 7bc] Figure 7bc shows the results of confirming that Brixelle treatment of a reconstructed human epidermis model (KeraSkin™) under inflammatory conditions (UVB irradiation) dose-dependently restored skin barrier function and suppressed inflammatory responses. [Figure 7bd]Figure 7b and 7d show the results of confirming that Brixelle treatment of a reconstructed human epidermis model (KeraSkin™) under inflammatory conditions (UVB irradiation) dose-dependently restored skin barrier function and suppressed inflammatory responses. [Figure 7c] Figure 7c shows the results of confirming that Brixelle treatment of a reconstructed human epidermis model (KeraSkin™) under inflammatory conditions (UVB irradiation) dose-dependently restored skin barrier function and suppressed inflammatory responses. [Figure 8a] Figure 8a shows the results of daily topical treatment with Brixelle for three weeks in a mouse model in which atopic dermatitis was induced by house dust mite antigen, confirming the dose-dependent therapeutic effect of Brixelle on atopic dermatitis. [Figure 8b] Figure 8b shows the results of daily topical treatment with Brixelle for three weeks in a mouse model in which atopic dermatitis was induced by house dust mite antigen, confirming the dose-dependent therapeutic effect of Brixelle on atopic dermatitis. [Figure 8c] Figure 8c shows the results of daily topical treatment with Brixelle for three weeks in a mouse model in which atopic dermatitis was induced by house dust mite antigen, confirming the dose-dependent therapeutic effect of Brixelle on atopic dermatitis. [Figure 8d] Figure 8d shows the results of daily topical treatment with Brixelle for three weeks in a mouse model in which atopic dermatitis was induced by house dust mite antigen, confirming the dose-dependent therapeutic effect of Brixelle on atopic dermatitis. [Figure 8e] Figure 8e shows the results of daily topical treatment with Brixelle for three weeks in a mouse model in which atopic dermatitis was induced with house dust mite antigen, confirming the dose-dependent therapeutic effect of Brixelle on atopic dermatitis. [Figure 9a]Figure 9a shows the results of daily topical treatment with Brixelle for three weeks in a mouse model of atopic dermatitis induced by house dust mite antigen, comparing its therapeutic effect with that of drugs used to treat atopic dermatitis (Elidel, Dexametasone, Eucrisa). [Figure 9b] Figure 9b shows the results of daily topical treatment with Brixelle for three weeks in a mouse model of atopic dermatitis induced by house dust mite antigen, comparing its therapeutic effect with that of drugs used to treat atopic dermatitis (Elidel, Dexametasone, Eucrisa). [Figure 9c] Figure 9c shows the results of daily topical treatment with Brixelle for three weeks in a mouse model of atopic dermatitis induced by house dust mite antigen, comparing its therapeutic effect with that of drugs used to treat atopic dermatitis (Elidel, Dexametasone, Eucrisa). [Figure 9d] Figure 9d shows the results of daily topical treatment with Brixelle for three weeks in a mouse model of atopic dermatitis induced by house dust mite antigen, comparing its therapeutic effect with that of drugs used to treat atopic dermatitis (Elidel, Dexametasone, Eucrisa). [Figure 9e] Figure 9e shows the results of daily topical treatment with Brixelle for three weeks in a mouse model of atopic dermatitis induced by house dust mite antigen, and comparing the therapeutic effect with that of drugs used to treat atopic dermatitis (Elidel, Dexametasone, Eucrisa). [Figure 10a] Figure 10a shows the results of daily topical treatment with Brixelle for 7 days in a mouse model of psoriasis induced by imiquimod (IMQ), confirming the therapeutic effect on psoriasis. [Figure 10b] Figure 10b shows the results of daily topical treatment with Brixelle for 7 days in a mouse model of psoriasis induced by imiquimod (IMQ), confirming the therapeutic effect on psoriasis. [Figure 10c]Figure 10c shows the results of daily topical treatment with Brixelle for 7 days in a mouse model of psoriasis induced by imiquimod (IMQ), confirming the therapeutic effect on psoriasis. [Figure 10d] Figure 10d shows the results of daily topical treatment with Brixelle for 7 days in a mouse model of psoriasis induced by imiquimod (IMQ), confirming the therapeutic effect on psoriasis. [Figure 11a] Figure 11a shows the results of a mouse model of contact dermatitis induced by DNFB (2,4-dinitrofluorobenzene) in which Brixelle was topically treated daily for five days, confirming its therapeutic effect on contact dermatitis. [Figure 11b] Figure 11b shows the results of a mouse model of contact dermatitis induced by DNFB (2,4-dinitrofluorobenzene) in which Brixelle was topically treated daily for five days, confirming its therapeutic effect on contact dermatitis. [Figure 11c] Figure 11c shows the results of a mouse model of contact dermatitis induced by DNFB (2,4-dinitrofluorobenzene) in which Brixelle was topically treated daily for five days, confirming its therapeutic effect on contact dermatitis. [Figure 11d] Figure 11d shows the results of a mouse model of contact dermatitis induced by DNFB (2,4-dinitrofluorobenzene) in which Brixelle was topically treated daily for five days, confirming its therapeutic effect on contact dermatitis. [Figure 12a] Figure 12a shows the results of alternative animal testing methods using a reconstructed human epidermis model (KeraSkin™) and a reconstructed human corneal-like epithelium model (MCTT HCE™), in which skin irritation tests, ocular mucosa irritation tests, skin micronucleus tests, skin sensitization tests, and skin phototoxicity tests were conducted, confirming that Brixelle is a substance with an extremely low risk of skin toxicity. [Figure 12b]Figure 12b shows the results of alternative animal testing methods using a reconstructed human epidermis model (KeraSkin™) and a reconstructed human corneal-like epithelium model (MCTT HCE™), in which skin irritation tests, ocular mucosa irritation tests, skin micronucleus tests, skin sensitization tests, and skin phototoxicity tests were conducted, confirming that Brixelle is a substance with an extremely low risk of skin toxicity. [Figure 12c] Figure 12c shows the results of alternative animal testing methods using a reconstructed human epidermis model (KeraSkin™) and a reconstructed human corneal-like epithelium model (MCTT HCE™), in which skin irritation tests, ocular mucosa irritation tests, skin micronucleus tests, skin sensitization tests, and skin phototoxicity tests were conducted, confirming that Brixelle is a substance with an extremely low risk of skin toxicity. [Figure 12da] Figure 12da shows the results of skin irritation tests, ocular mucosa irritation tests, skin micronucleus tests, skin sensitization tests, and skin phototoxicity tests using alternative animal testing methods using a reconstructed human epidermis model (KeraSkin™) and a reconstructed human corneal-like epithelium model (MCTT HCE™), confirming that Brixelle is a substance with an extremely low risk of skin toxicity. [Figure 12db] Figure 12db shows the results of skin irritation tests, ocular mucosa irritation tests, skin micronucleus tests, skin sensitization tests, and skin phototoxicity tests conducted using alternative animal testing methods using a reconstructed human epidermis model (KeraSkin™) and a reconstructed human corneal-like epithelium model (MCTT HCE™), confirming that Brixelle is a substance with an extremely low risk of skin toxicity. [Figure 12e] Figure 12e shows the results of alternative animal testing methods using a reconstructed human epidermis model (KeraSkin™) and a reconstructed human corneal-like epithelium model (MCTT HCE™), in which skin irritation tests, ocular mucosa irritation tests, skin micronucleus tests, skin sensitization tests, and skin phototoxicity tests were conducted, confirming that Brixelle is a substance with an extremely low risk of skin toxicity. DETAILED DESCRIPTION OF THE INVENTION

[0025] The present invention will be described in detail below.

[0026] The present invention relates to a pharmaceutical composition for preventing or treating an inflammatory disease, comprising a compound represented by Chemical Formula 1, a solvate thereof, or a pharmaceutically acceptable salt thereof.

[0027] [ka]

[0028] In the above formula 1, R1 and R4 are vinyl groups or methyl groups, R2 and R3 are methyl groups or vinyl groups, but not all of them need to be vinyl groups or methyl groups, and R5 is polyethylene glycol (PEG) or a derivative thereof.

[0029] The molecular weight of the polyethylene glycol or derivative thereof may be, but is not limited to, 500 to 10,000, 500 to 9,000, 500 to 8,000, 500 to 7,000, 500 to 6,000, or 500 to 5,000.

[0030] The polyethylene glycol or derivative thereof may be linear or branched.

[0031] For example, the polyethylene glycol derivatives include methoxy PEG, succinimide of PEG propionic acid, succinimide of PEG butanoic acid, branched PEG-NHS, PEG succinimidyl succinate, succinimide of carboxymethylated PEG, benzotriazole carbonate of PEG, PEG-glycidyl ether, PEG-oxycarbonylimidazole, PEG nitrophenyl carbonates, PEG-aldehyde, PEG succinimidyl carboxymethyl ester, and PEG succinimidyl ester. ester).

[0032] Specifically, the compound may be a compound represented by any one of the following chemical formulas 2 to 7.

[0033] [ka]

[0034] [ka]

[0035] [ka]

[0036] [ka]

[0037] [ka]

[0038] [ka]

[0039] The pharmaceutical composition of the present invention may include nanoparticles formed by self-assembly of the compound represented by Formula 1.

[0040] The size of the nanoparticles may be, but is not limited to, 1 nm to 5,000 nm, more specifically 50 nm to 1,000 nm, 50 nm to 500 nm, or 50 nm to 300 nm.

[0041] The inflammatory disease may be at least one selected from the group consisting of inflammatory skin diseases, osteoarthritis, hepatitis, pneumonia, keratitis, gastritis, nephritis, tuberculosis, bronchitis, pleuritis, peritonitis, spondylitis, pancreatitis, inflammatory bowel disease, urethritis, cystitis, inflammatory arteriosclerosis, sepsis, periodontitis, gingivitis, and autoinflammatory diseases, but is not limited thereto.

[0042] Examples of the inflammatory skin diseases include atopic dermatitis, contact dermatitis, psoriasis, seborrheic dermatitis, pruritus, parapsoriasis, urticaria, lichen planus, sunburn, radiodermatitis, erythema multiforme, erythema nodosum, granuloma annulare, keratosis pilaris, xeroderma, panniculitis, pyoderma gangrenosum, acne, rosacea, and lupus erythematosus. The condition may be at least one selected from the group consisting of erythematosus, pemphigus, diaper dermatitis, pityriasis rosea, alopecia areata, androgenic alopecia, vitiligo, and decubitus ulcer.

[0043] The causes of the inflammatory disease include, but are not limited to, immune system abnormalities, infections, external stimuli or injuries, environmental factors, genetic factors, etc. For example, the inflammatory disease may be caused by increased oxidative stress.

[0044] The term "oxidative stress" refers to the phenomenon in which an imbalance caused by excessive production of oxidizing substances such as reactive oxygen species (ROS) or a lack of antioxidants leads to damage or destruction of cells and tissues. Reactive oxygen species (ROS) are chemically reactive molecules containing oxygen atoms. They are oxygen compounds produced within living organisms and have strong oxidizing properties that attack biological tissues and damage cells. Reactive oxygen species are produced during the normal metabolic process of oxygen, but their concentration can rise rapidly due to environmental stress, causing cell damage.

[0045] Causes of excessive production of reactive oxygen species and oxidative stress include, but are not limited to, various internal and external causes such as improper diet, unbalanced nutrition, irregular lifestyle, excessive exercise, smoking, drinking, overwork, immune response, surgery, administration of anticancer drugs, exhaust fumes, heavy metals, radiation, ultraviolet rays, ultrasound, electromagnetic waves, and chemicals (detergents, pesticides, etc.).

[0046] In addition, the inflammatory skin diseases may be caused by dry skin, increased sensitivity, pigmentation, local inflammation of the skin, etc., which are caused by damage to the barrier function of the epidermis, stratum corneum, etc. due to oxidative stress.

[0047] The term "prevention" refers to any action of suppressing or delaying the onset of a related disease by administering the composition of the present invention. A person of ordinary skill in the art to which the present invention pertains would know that a related disease can be prevented by administering the composition of the present invention before the onset of symptoms or at the early stage of onset.

[0048] The term "treatment" refers to any action that improves or beneficially changes the symptoms of the relevant disease by administering the composition of the present invention, including alleviation or amelioration. A person of ordinary skill in the art to which the present invention pertains would be able to know the exact criteria for the disease and judge the degree of improvement, enhancement, and treatment by referring to materials presented by the Korean Medical Association, etc.

[0049] The term "pharmaceutically acceptable" refers to the property of not causing significant irritation to an individual, cell, tissue, etc. to which the compound or composition is administered, and not impairing the biological activity and physical properties of the compound.

[0050] The term "pharmaceutically acceptable salts" refers to salts prepared with certain compounds of the present invention and relatively non-toxic acids or bases. Pharmaceutically acceptable salts may be, for example, acid addition salts or metal salts.

[0051] Acid addition salts can be formed from inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, nitrous or phosphorous acids, and non-toxic organic acids such as aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates and alkanedioates, aromatic acids, aliphatic and aromatic sulfonic acids. The pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6- The alkyl esters may include dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, or mandalate.

[0052] The metal salt may be a sodium, potassium or calcium salt. Metal salts can be prepared using a base, for example, alkali metal or alkaline earth metal salts can be obtained by dissolving the compound in an excess amount of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salts, and evaporating and drying the filtrate.

[0053] The administration route of the pharmaceutical composition of the present invention may be oral administration or parenteral administration, including, but not limited to, intraneural, intravenous, intramuscular, intraperitoneal, subcutaneous, rectal, and topical administration. The administration route of the pharmaceutical composition of the present invention may be, for example, parenteral administration, specifically, topical administration such as skin, nasal, ophthalmic, ear drops, vaginal suppository, and rectal suppository, more specifically, administration by topical skin application.

[0054] The term "administration" refers to the introduction of a given substance into an individual by a suitable method. The term "individual" refers to animals, such as mice, livestock, including humans, that are suffering from or may develop the relevant disease.

[0055] The pharmaceutical composition of the present invention contains carriers, excipients and diluents, and can be formulated by a conventional method into oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions, but is not limited to these.

[0056] The carrier, excipient and diluent include, but are not limited to, lactose, dextrose, sucrose, dextrin, maltodextrin, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.When preparing the formulation, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc. that are commonly used in the art are used, but are not limited to these.

[0057] Solid preparations for oral administration include tablets, capsules, pills, granules, etc. The solid preparations can be prepared by mixing the composition with at least one or more excipients, such as sucrose, lactose, starch, gelatin, calcium carbonate, etc. In addition to the excipients, lubricants such as magnesium stearate and talc can also be used.

[0058] Liquid preparations for oral administration include aqueous solutions, suspensions, syrups, emulsions, etc. These liquid preparations may contain a variety of excipients, such as wetting agents, sweeteners, flavoring agents, preservatives, etc., in addition to simple diluents such as water and liquid paraffin.

[0059] Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Examples of suppository bases include witepsol, macrogol, tween 61, cocoa butter, laurin butter, and glycerogelatin.

[0060] Preparations for parenteral administration include, for example, sterile injectable preparations. Sterile injectable preparations may be aqueous or oily suspensions. Such suspensions can be formulated according to techniques well known in the art using appropriate dispersants, wetting agents (e.g., Tween 80), or suspending agents. Sterile injectable preparations may be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as mannitol, water, Ringer's solution, or isotonic sodium chloride solution. Generally, sterile, non-volatile oils can be used as solvents or suspending media, and any non-volatile oil with low irritation, such as synthetic mono- or diglycerides, can be used without limitation.

[0061] Preparations for topical administration during parenteral administration include topical preparations (liniments), eye drops, nasal drops, inhalants, vaginal preparations, suppositories, mouthwashes, etc. For example, when formulated as a topical preparation, it may contain adjuvants commonly used in topical skin preparations, such as fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic emulsifiers, nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, moisturizing agents, essential oils, dyes, pigments, hydrophilic active agents, lipophilic active agents, or lipid vesicles. The topical preparation may be in the form of, but is not limited to, an ointment, patch, gel, cream, or spray.

[0062] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to any medical treatment. The effective dose level can be determined based on factors including the patient's condition and weight, the type and severity of the disease, the activity of the drug, sensitivity to the drug, the administration time, administration route, and excretion rate, other factors well known in the medical field, and other factors. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with a conventional therapeutic agent for inflammatory diseases or oxidative stress-related skin diseases. The conventional therapeutic agent may be administered sequentially or simultaneously, and in single or multiple administrations. Taking all of the above factors into consideration, it is important to administer the minimum amount necessary to achieve maximum efficacy without side effects, and such an amount can be easily determined by one of ordinary skill in the art.

[0063] The effective dose of the pharmaceutical composition of the present invention varies depending on the age, sex, weight, etc. of the patient, and can be administered, for example, in an amount of 0.01 to 1,000 mg / kg / day, preferably 0.1 to 500 mg / kg / day, in one or several divided doses. However, the effective dose may increase or decrease depending on the route of administration, the number of doses, the severity of the disease, the age, the sensitivity to the drug, etc., and therefore the above dose does not in any way limit the scope of the present invention.

[0064] The present invention relates to a cosmetic composition containing a compound represented by Chemical Formula 1, a solvate thereof, or a cosmetically acceptable salt thereof.

[0065] [ka]

[0066] In the above formula 1, R1 and R4 are vinyl groups or methyl groups, R2 and R3 are methyl groups or vinyl groups, but not all of them need to be vinyl groups or methyl groups, and R5 is polyethylene glycol (PEG) or a derivative thereof.

[0067] The polyethylene glycol or derivative thereof may be within the ranges described above.

[0068] The compound may be a compound represented by any one of the above chemical formulas 2 to 7.

[0069] The cosmetic composition of the present invention may contain nanoparticles formed by self-assembly of the compound represented by Chemical Formula 1.

[0070] The nanoparticle content may be within the ranges mentioned above.

[0071] The cosmetic composition of the present invention may be for anti-inflammatory or anti-oxidative purposes.

[0072] The cosmetic composition of the present invention may also be used to prevent or improve skin inflammation caused by increased oxidative stress.

[0073] Skin inflammation caused by increased oxidative stress can occur due to the impairment of the barrier function of the epidermis, stratum corneum, etc., resulting in dry skin, increased sensitivity, pigmentation, localized inflammation of the skin, etc.

[0074] For example, skin inflammation caused by increased oxidative stress is a common skin condition that can be manifested as atopic dermatitis, contact dermatitis, psoriasis, seborrheic dermatitis, pruritus, parapsoriasis, urticaria, lichen planus, sunburn, radiodermatitis, erythema multiforme, erythema nodosum, granuloma annulare, keratosis pilaris, xeroderma, panniculitis, pyoderma gangrenosum, acne, rosacea, lupus erythematosus, and the like. The condition may be at least one selected from the group consisting of erythematosus, pemphigus, diaper dermatitis, pityriasis rosea, alopecia areata, androgenic alopecia, vitiligo, and decubitus ulcer.

[0075] The cosmetic composition of the present invention may also be used for moisturizing the skin, restoring the skin barrier function, or preventing skin aging.

[0076] Damage to the skin barrier can occur due to skin aging, ultraviolet rays, hormones, etc. Damage to the skin barrier can also be accompanied by skin inflammation caused by increased oxidative stress, such as the aforementioned atopic dermatitis and psoriasis.

[0077] Skin aging can be induced by ultraviolet rays, active oxygen, dryness, etc. Since the compound of Chemical Formula 1 contained in the cosmetic composition of the present invention can scavenge active oxygen, the cosmetic composition of the present invention can have the effect of preventing or repairing skin aging caused by active oxygen and damage to the skin barrier caused by skin aging.

[0078] The cosmetic composition of the present invention may be prepared in the form of a solution, topical ointment, cream, foam, nutritious lotion, softening lotion, pack, softening lotion, emulsion, makeup base, essence, soap, liquid cleanser, bath additive, sunscreen cream, sun oil, suspension, emulsion, paste, gel, lotion, powder, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch, or spray, but is not limited to these.

[0079] The cosmetic composition may contain one or more additives selected from aqueous additives such as purified water, stabilizers, emulsifiers, thickeners, moisturizers, liquid crystal film strengtheners, pH adjusters, antibacterial agents, water-soluble polymers, film-forming agents, sequestering agents, amino acids, organic amines, polymer emulsions, skin nutrients, antioxidants, antioxidant aids, preservatives, and fragrances; and oil-based additives such as fats and oils, waxes, hydrocarbon oils, higher fatty acid oils, higher alcohols, synthetic ester oils, and silicone oils.

[0080] The aqueous additive is not limited as long as it is a raw material commonly used in the field, and specific examples thereof include glycerin, dipropylene glycol, butylene glycol, pentylene glycol, methylpropanediol, sorbitol, diglycerin, erythritol, pentaerythritol, polybutylene glycol-10, polyglycerin-3, polyglycerin-4, polyglycerin-6, polyglycerin-10, polyglycerin-20, polyglycerin-40, sorbeth-5, and sorbeth-6. , Sorbeth-20, Sorbeth-30, Sorbeth-40, Inositol, Maltitol, Maltose, Mannan, Mannitol, Mannose, Lactitol, Lactose, Dihydroxypropyl PG-Glucoside, Dithiaoctanediol, Fructose, Glucamine, Methylglucamine, Glucose, 1,2,6-Hexanethiol, Methyl Gluceth-10, Methyl Gluceth-20, Ozonated Glycerin, Phytantriol, Thioglycerin, Threitol, Trimethylolpropane, Chlorpheniramine Examples of suitable surfactants include one or more selected from the group consisting of ciritol, EDTA, guar gum, quince seed, carrageenan, galactan, gum arabic, pectin, mannan, starch, xanthan gum, curdlan, methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, methylhydroxypropyl cellulose, chondroitin sulfate, dermatan sulfate, glycogen, heparan sulfate, hyaluronic acid, sodium hyaluronate, tragacanth gum, keratan sulfate, chondroitin, mucoitin sulfate, hydroxyethyl guar gum, carboxymethyl guar gum, dextran, keratosulfuric acid, locust bean gum, succinoglucan, caronic acid, chitin, chitosan, carboxymethyl chitin, agar, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium polyacrylate, polyethylene glycol, bentonite, methylparaben, propylparaben, phenoxyethanol, 1,2-hexanediol, and ethylhexylglycerin.

[0081] The oily additives are not limited as long as they are raw materials commonly used in the field, and examples thereof include liquid oils such as olive oil, camellia oil, jojoba oil, triglycerides, glycerin trioctanoate, and glycerin triisopalmitate, solid oils such as coconut oil, hydrogenated coconut oil, palm oil, hydrogenated oil, and hydrogenated castor oil, beeswax, candelilla wax, carnauba wax, lanolin, and jojoba wax. Examples of hydrocarbon oils include liquid paraffin, squalene, petrolatum, and microcrystalline wax. Examples of higher fatty acids include waxes such as lauric acid, myristic acid, palmitic acid, stearic acid, and behenic acid, cetyl alcohol, stearyl alcohol, behenyl alcohol, myristyl alcohol, and cetostearyl alcohol. Examples of synthetic ester oils include higher alcohols such as isopropyl myristate, cetyl octanoate, octyldodecyl myristate, isopropyl palmitate, hexyl laurate, myristyl myristate, cetyl lactate, isocetyl isostearate, neopentyl glycol dicaprate, ethylhexylglycerin, cetyl ethylhexanoate, ethylhexyl palmitate, and cetostearyl alcohol. Examples of synthetic ester oils include, but are not limited to, those selected from: linear silicone oils such as dimethylpolysiloxane, methylphenylpolysiloxane, and methylhydrogenpolysiloxane; and cyclic silicone oils such as dodecamethylcyclohexasiloxane, octamethylcyclotetrasiloxane, and decamethylcyclopentasiloxane.

[0082] The cosmetic composition of the present invention may further comprise a cosmetically acceptable carrier that is commonly blended in skin cosmetics commonly used in the art, such as, but not limited to, oil, water, surfactant, moisturizer, lower alcohol, thickener, chelating agent, colorant, preservative, fragrance, etc.

[0083] Depending on the formulation of the cosmetic composition of the present invention, various cosmetically acceptable carriers may be included.

[0084] When the cosmetic composition is formulated as an ointment, paste, cream, or gel, the carrier component may be an animal oil, a vegetable oil, a wax, a paraffin, a starch, tragacanth, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, silica, talc, zinc oxide, or a mixture thereof.

[0085] When the cosmetic composition is formulated as a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or a mixture thereof can be used as a carrier component. In particular, when the cosmetic composition is formulated as a spray, a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether can be further included.

[0086] When the cosmetic composition is formulated as a solution or emulsion, a solvent, solubilizer, or emulsifier can be used as the carrier component, and may be, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol fatty esters, polyethylene glycol, or fatty acid esters of sorbitan.

[0087] When the cosmetic composition is formulated as a suspension, the carrier component may be a liquid diluent such as water, ethanol, or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tragacanth.

[0088] When the formulation of the cosmetic composition is a surfactant-containing cleansing agent, the carrier component may be a fatty alcohol sulfate, a fatty alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyl taurate, a sarcosinate, a fatty acid amide ether sulfate, an alkylamidobetaine, a fatty alcohol, a fatty acid glyceride, a fatty acid diethanolamide, a vegetable oil, a lanolin derivative, or an ethoxylated glycerol fatty acid ester.

[0089] In addition to the above-mentioned compound and carrier component, the components contained in the cosmetic composition of the present invention may include components that are usually contained in cosmetic compositions, such as usual adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0090] The cosmetic composition may be used alone or in multiple applications, or may be used in multiple applications with other cosmetic compositions other than the composition of the present invention. The cosmetic composition of the present invention may be used according to a normal method of use, and the number of applications may be changed depending on the skin condition or preference of the user.

[0091] The present invention will be specifically described below with reference to examples.

[0092] Manufacturing example 1. Brixelle Manufacturing PEGylated bilirubin 3α of the following chemical formulas 2 to 7 (compounds 1 to 6, respectively) was synthesized by varying the molecular weight of PEG, and their molecular weights were analyzed (Figure 1). Using the synthesized PEGylated bilirubin 3α, its self-assembly, Brixelle, was produced using the following method.

[0093] [Table 1] JPEG2025525431000019.jpg197158

[0094] Brixelle was produced based on flow chemistry to produce uniformly sized nanoparticles.

[0095] Specifically, 10 L of citrate buffer solution was prepared by dissolving sodium citrate dihydrate and anhydrous citric acid in water, and the pH was confirmed to be 4.4. 9.9 L of the citrate buffer solution was then placed in a 30 L reactor. The solution was stirred at 100 to 1000 rpm while adjusting the temperature to maintain a temperature of 4 to 15°C.

[0096] Next, 1100 mL of acetonitrile was added dropwise to 504.4 g of PEGylated bilirubin 3α and stirred until completely dissolved. The solution was transferred to a 2 L graduated cylinder and acetonitrile was added to bring the total volume to 1650 mL. The solution was filtered through a 0.20 μm PVDF filter and transferred to a reactor. The mixture was stirred at 100-1000 rpm while adjusting the temperature to maintain a temperature of 4-15°C. The process was carried out under yellow light.

[0097] To formulate nanoparticles, 1650 mL of PEGylated bilirubin 3α solution was added to a citrate buffer solution. A 3.2 mm inner diameter tube was connected to a pump, and the PEGylated bilirubin 3α solution was pumped into the aqueous solution at a flow rate of 10–50 mL / min using the pump and tubing. After the dropwise addition of the PEGylated bilirubin 3α solution was completed, the mixture was stirred at 100–1000 rpm for 15 minutes at 4–15°C. This process was also carried out under yellow light. After stirring was complete, the reaction mixture was filtered through a 0.20 μm PVDF filter, transferred to a steel tray, and lyophilized to obtain Brixelle lyophilized powder. The completed Brixelle powder was redissolved in a portion of distilled water, and the formation of nanoparticles was monitored by dynamic light scattering (DLS) (Figures 2a and 2b).

[0098] 2. Preparation of Dil (Tetramethylindocarbocyanine Perchlorate)@Brixelle To load Brixelle Dil dye, the following was prepared:

[0099] 135 mg of PEGylated bilirubin 3α was dissolved in 6.16 mL of chloroform with 15 mg of Diluent. The solution was filtered through a 0.20 μm PTFE filter and then evaporated to a thin film, which was then dried under vacuum for 24 hours.

[0100] The dried reactant was dissolved in 14.6 mL of distilled water and then ultrasonicated in an ultrasonic cleaner at 20-30°C for 5-15 minutes to obtain a dispersion. The aqueous solution containing Dil@Brixelle was purified by filtering through a 0.45 μm PVDF filter and a 0.20 μm PVDF filter, and the filtrate was freeze-dried to obtain a freeze-dried powder of Dil@Brixelle.

[0101] The completed Dil@Brixelle powder was redissolved in a portion of distilled water, and the formation of Dil@Brixelle nanoparticles was confirmed by dynamic light scattering (DLS) (Figures 3a and 3b). The maximum absorbance of Brixelle was compared with that of Dil using a multi-mode plate reader to confirm the presence or absence of Dil dye loading (Figure 3c; the graph formed from wavelengths above 500 nm is the graph for Dil@Brixelle (High Concentration)).

[0102] Example The following experiments were carried out using Brixelle and Dil@Brixelle produced in Production Examples 1 and 2 above.

[0103] 1. Results of Brixelle uptake into skin cells under inflammatory conditions (1) Experimental method To confirm the uptake of Brixelle into skin cells under inflammatory conditions (stimulation with house dust mites (HDM) antigen), a human epidermal keratinocyte (HaCaT) cell line was used. HDM antigens are known to induce increased reactive oxygen species (ROS) production in immune and inflammatory cells associated with the Th2 immune response in atopic dermatitis and asthma, and are used as the primary antigenic stimulus in atopic dermatitis-induced cell models. To establish the inflammatory cell model, HaCaT cells were treated with 500 μg / mL HDM antigen for 24 hours. The prepared inflammatory cells were then dispensed into 24-well culture plates and treated with Brixelle (Dil@Brixelle) loaded with the fluorescent dye Dil (tetramethylindocarbocyanine perchlorate). The intracellular fluorescence expression was then monitored hourly for 24 hours using a MuviCyte real-time fluorescence microscope.

[0104] (2) Experimental results When Brixelle uptake into HaCaT cells under inflammatory conditions was compared, it was observed that Brixelle uptake was significantly increased in inflammatory cells stimulated with HDM antigen compared to normal cells not stimulated with HDM antigen. In particular, Brixelle uptake into inflammatory cells rapidly increased approximately 4 hours after Brixelle treatment, and after 24 hours it was confirmed to be approximately 6.2-fold higher than that of normal cells (Figures 4a and 4b).

[0105] 2. Results of Brixelle's ability to eliminate reactive oxygen species in skin cells and protect cells (1) Experimental method Compound 48 / 80 (C48 / 80) is known to mediate inflammatory responses and cell differentiation, as well as its role as a mast cell degranulator. In particular, C48 / 80 has been reported to be involved in skin inflammation through ROS generation in keratinocytes and mast cell degranulation. Therefore, in this study, HDM and C48 / 80 were used as antigenic and non-antigenic stimuli, respectively, in an inflammatory cell model. To evaluate Brixelle's ROS-scavenging ability in skin cells, HaCaT cells were first aliquoted into 96-well black plates. They were then treated with HDM and C48 / 80 at concentrations of 500 μg / mL and 20 μg / mL, respectively, to induce inflammation. They were also simultaneously treated with Brixelle (the test substance) and N-acetylcysteine (NAC) (the control substance) at various concentrations. After 24 hours, the culture medium was removed, and the cells were washed three times with sterile phosphate-buffered saline (PBS). Again, the cells were treated with 20 μM 2′,7′-dichlorofluorescein diacetate (DCF-DA) for 45 minutes to stain intracellular ROS, and the fluorescence intensity of the reaction solution was measured in the wavelength range λex=485 / λem=535 nm.

[0106] Next, to evaluate the ability of Brixelle to protect skin cells against pro-inflammatory stimuli, antigenic and non-antigenic stimuli were administered at a concentration that reduced HaCaT cell viability by 90% (90% cytotoxic concentration, CC). 90 ) (HDM = 2.5 mg / mL, C48 / 80 = 35 μg / mL). Simultaneously with the treatment of the pro-inflammatory stimuli, the test substance Brixelle and the control substance NAC were treated at different concentrations. After 24 hours, the culture medium was removed, washed three times with sterile PBS, and replaced with fresh culture medium. Then, 10 μL of Quanti-Max WST-8 solution, equivalent to 1 / 10 the volume of the culture medium, was dispensed into each well. After 4 hours of incubation, the absorbance of the reaction solution at 450 nm was measured to evaluate cell viability.

[0107] (2) Experimental results Brixelle's dose-dependent ROS scavenging effect was confirmed in inflammation-induced cell lines induced by antigenic or non-antigenic stimuli. In inflammation-induced cell lines induced by HDM, Brixelle's intracellular ROS scavenging ability (50% effective concentration, EC 50 ) was 78.8 μM, approximately 3.7 times higher than that of the comparative substance NAC (Fig. 5a). In the inflammation-induced cell line induced by C48 / 80, the EC 50 The value was 48.1 μM, which was approximately 6.2 times higher than that of NAC (FIG. 5b).

[0108] Brixelle demonstrated dose-dependent cytoprotective effects against both antigenic and non-antigenic stimuli in inflammatory cell lines. The EC values of Brixelle's cytoprotective effect in HDM-induced inflammatory cell lines were 50 The EC value of Brixelle was 93.9 μM, approximately 1.6-fold higher than that of NAC (Figure 5c). The EC value of Brixelle's cytoprotective activity was also significantly higher in the C48 / 80-induced inflammation-induced cell line. 50 The value was 67.0 μM, which was approximately 4.8 times higher than that of NAC (FIG. 5d).

[0109] These findings confirmed Brixelle's intracellular ROS scavenging effect in an inflammation-induced cell model, demonstrating its cytoprotective effect based on its strong antioxidant effect.

[0110] 3. Confirmation of Brixelle's ability to suppress inflammatory cytokines in skin cells (1) Experimental method Thymic stromal-derived lymphopoietin (TSLP), interleukin (IL)-25, and IL-33, produced by keratinocytes, are considered important mediators of Th2 immune responses in atopic dermatitis. We investigated the effect of Brixelle on the expression of representative inflammatory cytokines secreted from an inflammatory cell model induced by antigenic and non-antigenic stimuli. HaCaT cells were seeded into 96-well culture plates and treated with HDM and C48 / 80 at concentrations of 500 μg / mL and 20 μg / mL, respectively, as well as the test substance Brixelle and the control substance NAC at concentrations of 5, 50, and 500 μM. After 24 hours of incubation, cell supernatants were collected and subjected to enzyme-linked immunosorbent assay (ELISA) for each cytokine. The absorbance of the reaction solution was measured at 450 nm to quantify the amount of cytokine secreted by the inflammatory cell lines. Results are presented as mean ± standard error of the mean (SEM). Statistical analysis was performed using GraphPad. One-way ANOVA was used for statistical analysis, and statistical significance compared to the negative control group was confirmed using Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0111] (2) Experimental results The levels of three cytokines secreted from the inflammatory cell model induced by HDM and C48 / 80 stimuli showed a statistically significant increase compared to normal cells (p=0.0006, p<0.0001), demonstrating that the inflammatory cell model induced by these stimuli is suitable for confirming Brixelle's ability to regulate inflammatory cytokines.

[0112] In HDM-induced inflammatory cell lines, Brixelle treatment reduced the expression of all three cytokines in a concentration-dependent manner. Compared to the negative control, Brixelle treatment at 50 μM reduced TSLP by 26.4% and Brixelle treatment at 500 μM reduced TSLP by 50.1%, demonstrating a statistically significant difference from the negative control at 500 μM (Fig. 6a; p=0.0224). Compared to the negative control, Brixelle treatment at 50 μM reduced IL-25 by 47.1% and Brixelle treatment at 500 μM reduced IL-25 by 52.5%, demonstrating a statistically significant difference from the negative control at both concentrations (Fig. 6b; p=0.0054, p=0.0018). In the case of IL-33, a clear reduction of approximately 75.1% was observed in the Brixelle 500 μM treated group compared to the negative control group, which was confirmed to be a statistically significant difference (Figure 6c; p=0.001).

[0113] In the C48 / 80-induced inflammatory cell line, Brixelle treatment reduced the expression of all three cytokines in a concentration-dependent manner, demonstrating superior reduction compared to NAC at the same concentration. For TSLP, Brixelle treatment at 50 μM reduced TSLP by 31.3% and Brixelle treatment at 500 μM reduced TSLP by 38.3% compared to the negative control, with both treatment groups showing statistically significant differences from the negative control (Figure 6d; p=0.0143, p=0.0058). For IL-25, Brixelle treatment at 50 μM reduced IL-25 by 35.2% and 40% compared to the negative control, with the Brixelle treatment at 500 μM showing statistically significant differences from the negative control (Figure 6e; p=0.0231). In the case of IL-33, the Brixelle 50 μM treatment group showed a 21.1% reduction compared to the negative control group, and the Brixelle 500 μM treatment group showed a 55.9% reduction, showing a statistically significant difference from the negative control group in the Brixelle 500 μM treatment group (Figure 6f; p = 0.0120).

[0114] These results confirm that Brixelle, which has a strong antioxidant effect, has a superior mechanism of action in suppressing inflammatory responses in inflammatory cell lines compared to NAC, a conventionally known antioxidant.

[0115] 4. Brixelle's anti-inflammatory effect confirmed in a reconstructed human epidermis model (1) Experimental method A reconstructed human epidermis model (KeraSkin), a human skin tissue model TM The purpose of this study was to investigate the anti-inflammatory ability of Brixelle in reconstructed human epidermis models (0.6 cm). 2 The cells were then allowed to stabilize in a 37°C, 5% CO2 cell incubator for 22±2 hours. To induce an inflammatory response in the reconstructed human epidermis model, the culture plate was placed in an ultraviolet (UVB) irradiator and irradiated with 300 mJ / cm2. 2 The reconstructed human epidermis model was then exposed to UV light for 10 min. After inflammation induction, Brixelle 80 (400 μM) was slowly applied to the top center of the model using a pipette and spread evenly across the entire surface. After 8 hours of treatment with the test substance in the cell culture vessel, the test substance was removed using Dulbecco's PBS (DPBS). The cells were then cultured for 40 hours, and transepithelial electrical resistance (TEER) and the expression of inflammatory cytokines in the culture medium were measured. Histological staining was performed after the test to confirm Brixelle's ability to inhibit skin barrier damage.

[0116] To evaluate the function and strength of the skin barrier, TEER was measured using an electrical resistance system (ERS-2, Millipore). To confirm changes in the inflammatory response, tumor necrosis factor-α (TNF-α), IL-6, IL-1α, and TSLP were measured using ELISA. Furthermore, hematoxylin and eosin (H&E) staining was performed to confirm histological changes in the reconstructed human epidermis model, and filaggrin immunohistochemistry was used to evaluate skin barrier function. TEER and inflammatory cytokine results are expressed as mean ± standard deviation (SD). Statistical analysis was performed using the GraphPad program. One-way ANOVA was used for statistical analysis, and statistical significance between test groups was confirmed using Tukey's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0117] (2) Experimental results The epithelial cell layer has developed tight junctions, a type of junction between cells, forming a membranous physicochemical barrier. It is known that skin barrier function is impaired when an inflammatory response occurs in the skin. TEER, an index measuring the strength of tight junctions, decreases as barrier function deteriorates. In this experiment, an inflammatory response was induced in a reconstructed human epidermis model by UV irradiation, and the effect of Brixelle on protecting skin barrier function was examined by analyzing changes in TEER. After UV irradiation, the TEER of the reconstructed human epidermis model decreased compared to the negative control group. However, in the Brixelle-treated group, TEER significantly recovered in a concentration-dependent manner in measurements taken 24 and 48 hours later (Figure 7a).

[0118] It is known that pro-inflammatory cytokines are expressed at higher levels in various inflammatory diseases of the skin than in normal skin tissue. In this experiment, we analyzed TNF-α, IL-6, and IL-1α. We also measured TSLP, which has been shown to be involved in allergies and cutaneous immune-mediated diseases and plays an important role in regulating inflammatory responses (Figure 7b (7ba-7bd)).

[0119] The four cytokines were measured by ELISA. TNF-α expression was significantly increased in the induced-inflammatory group compared to the negative control group. However, Brixelle 80 and 400 μM treatments showed a significant concentration-dependent decrease in expression (negative control vs. UVB: 16.8 ± 2.6 pg / mL vs. 61.1 ± 1.2 pg / mL, p<0.0001; UVB vs. Brixelle 80 μM: 61.1 ± 1.2 pg / mL vs. 14.7 ± 0.3 pg / mL, p<0.0001; UVB vs. Brixelle 400 μM: 61.1 ± 1.2 vs. 15.6 ± 0.0 pg / mL, p<0.0001).

[0120] When the expression level of IL-6 was examined, the p values for the comparison between the negative control group and the inflammation-induced group, and between the inflammation-induced group and the Brixelle 400μM treatment group were 0.0004 and 0.0034, respectively, and a significant decrease in expression was confirmed when Brixelle was treated compared to the inflammation-induced group (negative control vs UVB: 3.2±0.0 vs 30.0±2.3, p=0.0004; UVB vs Brixelle 400μM: 30.0±2.3 vs 15.0±2.3, p=0.0034).

[0121] When the expression levels of IL-1α in the culture medium were compared between the IL-1α negative control group and the inflammation-induced group, and between the inflammation-induced group and the Brixelle 80μM and 400μM treatment groups, the p-values between the groups were all 0.0058 (p<0.01) (negative control vs UVB: 3.9±0.0 vs 14.1±3.0, p=0.0058; UVB vs Brixelle 80μM: 14.1±3.0 vs 3.9±0.0, p=0.0058; UVB vs Brixelle 400μM: 14.1±3.0 vs 3.9±0.0, p=0.0058).

[0122] Furthermore, the expression levels of TSLP, which regulates the inflammatory response, were 3.3 ± 0.0, 23.7 ± 1.9, 10.1 ± 1.6, and 9.1 ± 1.4 pg / mL in the negative control, induced inflammation, and Brixelle 80 μM and 400 μM treatment groups, respectively. Comparison of the expression levels in the induced inflammation group with each group revealed a statistically significant decrease in TSLP expression after Brixelle treatment after inflammation induction (p<0.001, p<0.01, and p<0.01, respectively) (negative control vs. UVB, p=0.0004; UVB vs. Brixelle 80 μM, p=0.0020; UVB vs. Brixelle 400 μM, p=0.0015).

[0123] We investigated the histopathology and expression of inflammation and skin barrier function markers in the reconstructed human epidermis model using H&E staining and filaggrin immunohistochemical staining. Among the skin barrier function markers, filaggrin is an essential factor for regulating epidermal homeostasis and, as a component of the lipid envelope within the stratum corneum, is responsible for skin moisturization and barrier function. H&E staining confirmed that the reconstructed human epidermis model is composed of four layers (stratum corneum, granular layer, stratum spinosum, and basal layer) similar to real skin. Inducing inflammation with UV irradiation reduced the number of stained cell nuclei compared to the negative control group, confirming the peeling of the normally layered stratum corneum. In contrast, the number of stained cell nuclei in the reconstructed human epidermis model treated with Brixelle remained unchanged compared to the inflammation-induced group, and the stratum corneum displayed a normal layered morphology. Immunohistochemical staining confirmed the expression of filaggrin in the granular and stratum corneum layers of the reconstructed human epidermis model. In the inflammation-induced group, UV irradiation reduced filaggrin expression, but Brixelle treatment inhibited this reduction in filaggrin expression (Figure 7c).

[0124] Summarizing the above results, Brixelle has been proven to effectively suppress inflammatory responses induced by UV irradiation in human skin tissue models, restore damaged skin barrier function, and contribute to histological improvement.

[0125] 5. Brixelle's efficacy in a mouse model of induced atopic dermatitis (1) (1) Experimental method A mouse model of HDM-induced atopic dermatitis was established by applying HDM antigen ointment to the dorsal and ear skin of 6-10-week-old NC / Nga female mice seven times on days 0, 7, 10, 14, 17, 21, and 24. A normal group (normal group) was used, and a negative control group (no treatment group) and a vehicle group (vehicle group) were used. A positive control group (0.1% dexamethasone, 62.5 mg / mouse) was used. The efficacy of the test substance, Brixelle (7.5, 15, and 30 mg / kg), was evaluated daily from day 7 onward. Clinical symptoms were assessed during the induction of atopic dermatitis, and skin tissue was collected for further analysis after the survival study.

[0126] To quantify the clinical symptoms of atopic dermatitis, clinical scores were measured on a 3-point Likert scale for four clinical indicators: 1) erythema / hemorrhage, 2) scarring / dryness, 3) edema, and 4) excoriation / erosion. H&E staining was performed to observe histological changes in the skin due to atopic dermatitis. Dihydroethidium (DHE) staining was performed to measure intradermal ROS. Toluidine blue and Congo red staining were performed to identify infiltrated mast cells and eosinophils, respectively. Graphs show mean ± SEM, and statistical analysis was performed using the GraphPad program. For statistical analysis, one-way analysis of variance (one-way ANOVA) was used, and statistical significance with the vehicle group was confirmed using Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0127] (2) Experimental results When the clinical symptoms were confirmed at the end of the survival test, it was found that the induction of atopic dermatitis resulted in flushing, edema, and keratin formation, and secondary skin damage due to itching was clearly observed. Clinical evaluation revealed that the groups treated with dexamethasone and Brixelle, both approved drugs, showed statistically significant improvements in clinical symptoms compared to the vehicle group, and Brixelle's effect was concentration-dependent (Figure 8a. Vehicle vs. Dexamethasone: 9.44±0.77 vs. 4.90±0.76, p<0.0001; Vehicle vs. Brixelle 7.5mg / kg: 9.44±0.77 vs. 5.90±0.47, p<0.0001; Vehicle vs. Brixelle 15mg / kg: 9.44±0.77 vs. 5.05±0.32, p<0.0001; Vehicle vs. Brixelle 30mg / kg: 9.44±0.77 vs. 4.40±0.48, p<0.0001).

[0128] Histological changes in the skin were confirmed by H&E staining, and an increase in epidermal thickness and hyperkeratosis were observed following treatment with HDM antigen ointment. Increased vascularization within the dermis, tissue edema, and inflammatory cell infiltration were also confirmed. The dexamethasone- and Brixelle-treated groups showed a decrease in epidermal thickness and hyperkeratosis compared to the vehicle group (Figure 8b).

[0129] To confirm whether Brixelle's antioxidant properties are effective in improving the symptoms of atopic dermatitis, we examined the expression of ROS in skin tissue. When ROS levels in skin tissue were measured by DHE staining, strong DHE staining was observed in the epidermis and dermis of the negative control group compared to the normal group, while reduced DHE staining intensity was observed in the dexamethasone-treated and Brixelle-treated groups (Figure 8c).

[0130] Additional staining was performed to confirm the infiltration of mast cells and eosinophils, inflammatory cells known to be associated with the symptoms of atopic dermatitis. Mast cells secrete histamine, and eosinophils secrete cytotoxic substances, and are therefore immune cells involved in allergic symptoms. Mast cell infiltration was quantified by toluidine blue staining, and statistically significant decreases in mast cell infiltration were observed in the dexamethasone- and Brixelle-treated groups compared with the vehicle group (Fig. 8d; Vehicle vs. Dexamethasone, 52.69±2.19 vs. 27.38±3.19, p<0.0001; Vehicle vs. Brixelle 7.5 mg / kg, 52.69±2.19 vs. 32.19±2.72, p<0.0001; Vehicle vs. Brixelle 15 mg / kg, 52.69±2.19 vs. 29.81±1.11, p<0.0001; Vehicle vs. Brixelle 30 mg / kg, 52.69±2.19 vs. 23.19±1.57, p<0.0001). Eosinophil infiltration was also quantified by Congo red staining, and statistically significant decreases in eosinophil infiltration were observed in the dexamethasone- and Brixelle-treated groups compared with the vehicle group (Figure 8e; Vehicle vs. Dexamethasone, 35.50±1.37 vs. 18.88±1.64, p<0.0001; Vehicle vs. Brixelle 7.5 mg / kg, 35.50±1.37 vs. 21.56±1.44, p<0.0001; Vehicle vs. Brixelle 15 mg / kg, 35.50±1.37 vs. 20.88±1.55, p<0.0001; Vehicle vs. Brixelle 30 mg / kg, 35.50±1.37 vs. 17.56±0.92, p<0.0001).

[0131] These results confirm that Brixelle, a powerful antioxidant, dose-dependently prevents the clinical symptoms, histological changes in the skin, and inflammatory responses associated with atopic dermatitis.

[0132] 6. Brixelle's efficacy in a mouse model of induced atopic dermatitis (2) (1) Experimental method A mouse model of HDM-induced atopic dermatitis was established by applying HDM antigen ointment to the back and ear skin of 6- to 10-week-old NC / Nga female mice seven times on days 0, 7, 10, 14, 17, 21, and 24. A normal group (no HDM antigen application), a negative control group (no treatment group) in which no additional substances were applied during the induction of atopic dermatitis, and a vehicle group (vehicle group) were also included. Starting on day 7, Brixelle (30 mg / kg) and approved drugs Elidel (1% pimecrolimus, 62.5 mg / mouse), Maxidex (0.1% dexamethasone, 62.5 mg / mouse), and Eucrisa (2% crisaborole, 62.5 mg / mouse) were applied daily to evaluate their efficacy. Clinical symptoms were assessed during the induction of atopic dermatitis, and skin tissues were secured for further analysis after the survival test was completed.

[0133] To quantify the clinical symptoms of atopic dermatitis, clinical scores were measured on a 3-point Likert scale for four clinical indicators: 1) erythema / hemorrhage, 2) scarring / dryness, 3) edema, and 4) excoriation / erosion. H&E staining was performed to observe histological changes in the skin due to atopic dermatitis. Dihydroethidium (DHE) staining was performed to measure intracutaneous ROS. Toluidine blue and Congo red staining were performed to identify infiltrated mast cells and eosinophils, respectively. ELISA was performed on skin tissue to measure IL-4 and IL-13, inflammatory mediators known to be highly associated with atopic dermatitis. The graphed results are expressed as mean ± SEM, and statistical analysis was performed using the GraphPad program. One-way ANOVA was used for statistical analysis, and statistical significance compared to the vehicle group was confirmed using Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0134] (2) Experimental results Clinical symptoms at the end of the survival study showed that induction of atopic dermatitis resulted in flushing, edema, and keratin formation, as well as clear secondary skin damage due to itching. Clinical evaluation revealed that Brixelle and two approved drugs (Dexamethasone and Eucrisa) showed statistically significant improvements in clinical symptoms compared to the vehicle group (Figure 9a. Vehicle vs. Dexamethasone: 9.58±0.64 vs. 3.75±0.99, p<0.0001; Vehicle vs. Eucrisa: 9.58±0.64 vs. 3.25±0.67, p<0.0001; Vehicle vs. Brixelle: 9.58±0.64 vs. 6.00±0.52, p=0.0034).

[0135] Histological changes in the skin were confirmed by H&E staining, and an increase in epidermal thickness and hyperkeratosis were observed following treatment with HDM antigen ointment. Furthermore, increased vascularization within the dermis, tissue edema, and inflammatory cell infiltration were confirmed. Compared to the vehicle group, Brixelle and two approved drugs (Dexamethasone and Eucrisa) treatment groups showed a decrease in epidermal thickness and hyperkeratosis (Figure 9b).

[0136] To confirm whether Brixelle's antioxidant properties are effective in improving the symptoms of atopic dermatitis, we examined the expression of ROS in skin tissue. When ROS levels in skin tissue were measured by DHE staining, strong DHE staining was observed in the epidermis and dermis of the negative control group compared to the normal group, while decreased DHE staining intensity was observed in the Brixelle and two approved drug (Dexamethasone and Eucrisa) treatment groups (Figure 9c).

[0137] Additional staining was performed to confirm the infiltration of mast cells and eosinophils, inflammatory cells known to be associated with the symptoms of atopic dermatitis. Mast cells secrete histamine, and eosinophils secrete cytotoxic substances, and are therefore immune cells involved in allergic symptoms. Mast cell infiltration was quantified using toluidine blue staining. A statistically significant decrease in mast cell infiltration was confirmed in the Brixelle and two approved drug (Dexamethasone and Eucrisa) treatment groups compared to the vehicle group (Figure 9d; Vehicle vs. Dexamethasone, 56.81±1.99 vs. 20.69±5.25, p<0.0001; Vehicle vs. Eucrisa, 56.81±1.99 vs. 20.31±2.55, p<0.0001; Vehicle vs. Brixelle, 56.81±1.99 vs. 27.69±1.94, p<0.0001). Furthermore, eosinophils were quantified using Congo red staining, and a statistically significant decrease in eosinophil infiltration was confirmed in the Brixelle and two approved drug (Dexamethasone and Eucrisa) treatment groups compared to the vehicle group (Figure 9e, Vehicle vs. Dexamethasone, 40.06±5.95 vs. 10.25±2.56, p<0.0001; Vehicle vs. Eucrisa, 40.06±5.95 vs. 10.31±2.49, p<0.0001; Vehicle vs. Brixelle, 40.06±5.95 vs. 21.69±2.77, p=0.0027).

[0138] These results indicate that Brixelle, as a potent antioxidant, has a mechanism of action to prevent the clinical symptoms, histological changes in the skin, and inflammatory responses of atopic dermatitis, similar to the approved drugs Elidel, Dexamethasone, and Eucrisa.

[0139] 7. Brixelle's efficacy confirmed in a mouse model of psoriasis (1) Experimental method Aldara was applied to the dorsal skin of 8-week-old C57BL / 6 male mice. TMA mouse model of IMQ-induced psoriasis was created by applying imiquimod (IMQ) cream to the skin seven times from day 0 to day 6. A normal group (normal group) received no IMQ cream; a negative control group (no treatment group) and a vehicle group (vehicle group) received no additional treatment during the psoriasis induction process; and a positive control group (clobetasol propionate group, 0.05% clobetasol propionate, 62.5 mg / mouse) received the approved drug Betabate. The test substance, Brixelle (30 mg / kg), was applied to the skin daily from day 0 to evaluate its efficacy. Clinical symptoms were evaluated during the psoriasis induction process, and skin tissue was obtained after the survival study for further analysis.

[0140] To quantify the clinical symptoms of psoriasis, three clinical indicators were measured: 1) erythema (redness), 2) scaling, and 3) skin thickness. Clinical scores were measured on a 4-point scale (modified psoriasis area and severity index, PASI). H&E staining was performed to observe histological changes in psoriasis-related skin, and DHE staining was performed to measure intradermal reactive oxygen species (ROS) in skin tissue. RT-PCR and ELISA were performed on skin tissue to measure IL-17 and IL-23, inflammatory mediators known to be highly associated with psoriasis. Graph results are presented as mean ± SEM. Statistical analysis was performed using the GraphPad program. One-way ANOVA was used for statistical analysis, and statistical significance compared with the vehicle group was confirmed using Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0141] (2) Experimental results Clinical symptoms at the end of the survival study were clearly observed, including flushing, edema, and keratinization due to the induction of psoriasis. Clinical evaluation revealed that the Clobetasol propionate and Brixelle treatment groups showed statistically significant improvements in clinical symptoms compared to the vehicle group (Figure 10a. Vehicle vs. Clobetasol propionate: 10.50±0.47 vs. 7.25±0.21, p<0.0001; Vehicle vs. Brixelle 30mg / kg: 10.50±0.47 vs. 6.00±0.22, p<0.0001).

[0142] Histological changes in the skin were confirmed by H&E staining, and it was observed that IMQ treatment increased epidermal thickness and hyperkeratosis. In the Brixelle-treated group, a decrease in epidermal thickness and hyperkeratosis was observed compared to the vehicle group (Figure 10b).

[0143] To confirm whether Brixelle's antioxidant properties are effective in improving psoriasis symptoms, we examined the expression of ROS in skin tissue. When ROS levels in skin tissue were measured by DHE staining, strong DHE staining was observed in the epidermis and dermis of the negative control group compared to the normal group, while decreased DHE staining intensity was observed in the Brixelle and positive treatment groups (Figure 10c).

[0144] IL-23 and IL-17, immune mediators associated with the Th1 and Th17 immune responses, are key mechanisms in the development of psoriasis symptoms. Their involvement in the initiation, maintenance, and amplification of Th1 and Th17 immune responses makes them targets for psoriasis therapeutics. Macrophages infiltrating the skin tissue of psoriasis patients express IL-23, which induces γδ T cells to express IL-17. Therefore, we analyzed the expression of IL-23 and IL-17 in skin tissue. Although the Brixelle treatment group did not show statistically significant reductions in IL-23 mRNA expression compared with the vehicle group, it demonstrated a similar reduction effect to the approved drug clobetasol propionate (Figure 10d). Brixelle demonstrated a statistically significant reduction in IL-17 protein expression compared to the vehicle group (Figure 10d. Vehicle vs Brixelle 30 mg / kg, 354 ± 36.5 vs 203 ± 15.2, P = 0.0378).

[0145] These results indicate that Brixelle, as a potent antioxidant, has a mechanism of action similar to that of the approved drug clobetasol propionate in preventing the clinical symptoms, histological changes in the skin, and inflammatory responses of psoriasis.

[0146] 8. Brixelle's efficacy in a mouse model of contact dermatitis (1) Experimental method A mouse model of DNFB-induced contact dermatitis was established using 6- to 10-week-old C57BL / 6 NC / Nga male mice and mouse 2,4-dinitrofluorobenzene (DNFB). Sensitization was achieved by applying a 0.5% DNFB solution (acetone:olive oil = 4:1) to the dorsal skin on days 0 and 1, followed by application of a 0.3% DNFB solution to the ear skin on day 7. A normal group (normal group) was not treated with DNFB solution. Negative control groups (no treatment group, no additional substance was applied during the contact dermatitis induction process) and a vehicle-treated group (vehicle group) were used. A positive control group (0.1% dexamethasone, 62.5 mg / mouse) was also used. The efficacy of the test substance, Brixelle (30 mg / kg), was evaluated daily from day 0 onward. Clinical symptoms were assessed during the induction of contact dermatitis, and skin tissues were secured for further analysis after the survival test was completed.

[0147] Ear thickness was measured to assess edema, a clinical symptom of contact dermatitis. H&E staining was performed to observe histological changes in the skin due to contact dermatitis, and DHE staining was performed to measure ROS in the skin tissue. RT-PCR was performed on skin tissue to measure TNF-α and IFN-γ, inflammatory mediators known to be highly associated with contact dermatitis. Results are presented as mean ± SEM, and statistical analysis was performed using the GraphPad program. One-way ANOVA was used for statistical analysis, and statistical significance compared to the vehicle group was confirmed using Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0148] (2) Experimental results Clinical signs at the end of the survival study clearly indicated edema due to induced contact dermatitis. Measurement of ear skin thickness revealed a statistically significant effect in the Brixelle-treated group compared to the vehicle-treated group (Figure 11a. Vehicle vs. Dexamethasone, 0.94±0.01 vs. 0.67±0.02, p<0.0001; Vehicle vs. Brixelle 30 mg / kg, 0.94±0.01 vs. 0.71±0.01, p<0.0001).

[0149] Histological changes in the ear skin were confirmed by H&E staining, which revealed an increase in skin thickness and tissue edema. The Brixelle-treated group showed a decrease in skin thickness and tissue edema compared to the vehicle group (Figure 11b).

[0150] To confirm whether Brixelle's antioxidant properties are effective in improving the symptoms of contact dermatitis, we examined the expression of ROS in ear skin tissue. When ROS levels in skin tissue were measured using DHE staining, strong DHE staining was observed in the negative control group compared to the normal group, while decreased DHE staining intensity was observed in the Brixelle-treated and positive-treated groups (Figure 11c).

[0151] TNF-α and IFN-γ, immune mediators associated with the Th1 immune response, an important mechanism in the manifestation of contact dermatitis symptoms, are involved in the induction of Th1 immune responses and the amplification of inflammation. RT-PCR was performed to compare the mRNA expression of TNF-α and IFN-γ in skin tissue. Although no statistically significant differences were observed in the Brixelle-treated group compared to the vehicle group, the Brixelle-treated group demonstrated a similar reduction in mRNA expression to that of the approved drug dexamethasone (Figure 11d).

[0152] These results indicate that Brixelle, as a potent antioxidant, has a mechanism of action similar to that of the approved drug dexamethasone in preventing the clinical symptoms, histological changes in the skin, and inflammatory responses associated with contact dermatitis.

[0153] 9. Brixelle skin toxicity confirmation results (1) Experimental method Reconstructed human epidermis model (KeraSkin TM ) and reconstructed human corneal epithelium model (MCTT HCE TM The purpose of this study was to investigate the effects of Brixelle on skin tissue using a reconstructed human epidermis model and a reconstructed human cornea-like epithelium model, which are artificial tissues that mimic human skin and cornea, respectively. Brixelle was then treated with three concentrations (10, 25, and 50 mM). A skin irritation test was then performed to confirm Brixelle's potential for skin irritation, an ocular mucosa irritation test to confirm its potential for ocular irritation, a skin sensitization test to confirm its potential for activating the skin's immune system, a skin phototoxicity test to confirm its potential for UVA-induced transformation into a skin irritant, and a skin micronucleus test to confirm its potential for genotoxicity in skin tissue.

[0154] (2) Experimental results In a skin irritation test to determine whether a test substance has the potential to irritate the skin, Brixelle demonstrated a cell viability of 50% or more up to a concentration of 50 mM, confirming its non-irritating nature (Figure 12a).

[0155] In an ocular mucosa irritation test to determine whether a test substance has the potential to irritate the ocular mucosa, Brixelle demonstrated a cell viability of 35% or more up to a concentration of 50 mM, confirming its non-irritating properties (Figure 12b).

[0156] In a skin micronucleus test to determine whether a test substance has the potential to cause genotoxicity in skin tissue, Brixelle did not induce significant micronuclei at concentrations up to 50 mM compared to the negative control group, and the lack of a concentration-dependent increase in micronuclei induction frequency confirmed its negative genotoxicity (Figure 12c).

[0157] In a skin sensitization test to determine whether the test substance has the potential to activate the skin's immune system, we evaluated whether the expression of CD54 and CD86 on the cell surface increased during the differentiation process of THP-1 cells, an immune cell-derived cell line, cultured with a reconstructed human epidermis model. The results showed that Brixelle reduced the expression of CD54 and CD86 on THP-1 cells cultured with Brixelle up to a concentration of 50 mM by 200 RFI and less than 120 RFI, respectively, compared to the negative control, confirming negative skin sensitization potential (Figures 12d (12da and 12db)).

[0158] In a skin phototoxicity test to determine whether the test substance could be transformed into a skin irritant by UVA, Brixelle showed a difference in cell viability of less than 30% before and after UVA irradiation up to a concentration of 50 mM, confirming negative skin phototoxicity (Figure 12e).

[0159] These results confirm that Brixelle, a powerful antioxidant, does not irritate the skin or ocular mucosa, does not activate the skin's immune system, does not react with UV rays to become an irritant, and does not cause genotoxicity.

Claims

1. A pharmaceutical composition for preventing or treating an inflammatory disease, comprising a compound represented by Chemical Formula 1, a solvate thereof, or a pharmaceutically acceptable salt thereof. 【Chemical 1】 (In the above chemical formula 1, R １ and R ４ is a vinyl group or a methyl group, and R ２ and R ３ are methyl groups or vinyl groups, but not all of them need to be vinyl groups or methyl groups; R ５ is polyethylene glycol (PEG) or a derivative thereof.

2. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the compound is a compound represented by any one of chemical formulas 2 to 7. 【Chemistry 2】 【Chemistry 3】 【Chemistry 4】 【Chemistry 5】 【Chemistry 6】 【Chemistry 7】

3. 10. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, comprising nanoparticles formed by self-assembly of the compound represented by Chemical Formula 1.

4. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 3, wherein the nanoparticles have a size of 1 nm to 5,000 nm.

5. The polyethylene glycol derivatives include methoxy PEG, succinimide of PEG propionic acid, succinimide of PEG butanoic acid, branched PEG-NHS, PEG succinimidyl succinate, succinimide of carboxymethylated PEG, and benzotriazole carbonate of PEG.

2. The pharmaceutical composition for the prevention or treatment of an inflammatory disease according to claim 1, wherein the PEG-100 hydroxybenzoate is selected from the group consisting of PEG, PEG-glycidyl ether, PEG-oxycarbonylimidazole, PEG nitrophenyl carbonates, PEG aldehyde, PEG succinimidyl carboxymethyl ester, and PEG succinimidyl ester.

6. 2. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the inflammatory disease is at least one selected from the group consisting of inflammatory skin diseases, osteoarthritis, hepatitis, pneumonia, keratitis, gastritis, nephritis, tuberculosis, bronchitis, pleuritis, peritonitis, spondylitis, pancreatitis, inflammatory bowel disease, urethritis, cystitis, inflammatory arteriosclerosis, sepsis, periodontitis, gingivitis, and autoinflammatory diseases.

7. The inflammatory skin diseases include atopic dermatitis, contact dermatitis, psoriasis, seborrheic dermatitis, pruritus, parapsoriasis, urticaria, lichen planus, sunburn, radiation dermatitis, erythema multiforme, erythema nodosum, granuloma annulare, and keratosis pilaris. Pilaris, Xeroderma, Panniculitis, Pyoderma gangrenosum, Acne, Rosacea, Lupus erythematosus, Pemphigus, Diaper dermatitis, Pityriasis rosea, Alopecia areata, Androgenic alopecia, Vitiligo, and Decubitus 7. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 6, wherein the compound is at least one selected from the group consisting of:

8. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the inflammatory disease is caused by increased oxidative stress.

9. A cosmetic composition comprising a compound represented by Chemical Formula 1, a solvate thereof, or a cosmetically acceptable salt thereof. 【Chemistry 8】 (In the above chemical formula 1, R １ and R ４ is a vinyl group or a methyl group, and R ２ and R ３ are methyl groups or vinyl groups, but not all of them need to be vinyl groups or methyl groups; R ５ is polyethylene glycol (PEG) or a derivative thereof.

10. The cosmetic composition according to claim 9, wherein the compound is a compound represented by any one of chemical formulas 2 to 7. 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】

11. The cosmetic composition according to claim 9, comprising nanoparticles formed by self-assembly of the compound represented by Chemical Formula 1.

12. The cosmetic composition according to claim 11, wherein the nanoparticles have a size of 1 nm to 5,000 nm.

13. The polyethylene glycol derivatives include methoxy PEG, succinimide of PEG propionic acid, succinimide of PEG butanoic acid, branched PEG-NHS, PEG succinimidyl succinate, succinimide of carboxymethylated PEG, and benzotriazole carbonate of PEG.

10. The cosmetic composition according to claim 9, wherein the PEG-10 alkyl acrylate or PEG-11 alkyl acrylate is selected from the group consisting of PEG, PEG-glycidyl ether, PEG-oxycarbonylimidazole, PEG nitrophenyl carbonates, PEG aldehyde, PEG succinimidyl carboxymethyl ester, and PEG succinimidyl ester.

14. The cosmetic composition according to claim 9, wherein the cosmetic composition is for anti-inflammatory or anti-oxidative use.

15. The cosmetic composition according to claim 9, which is for preventing or improving skin inflammation caused by increased oxidative stress.

16. Skin inflammation caused by increased oxidative stress is known to cause a variety of skin conditions, including atopic dermatitis, contact dermatitis, psoriasis, seborrheic dermatitis, pruritus, parapsoriasis, urticaria, lichen planus, sunburn, radiation dermatitis, erythema multiforme, erythema nodosum, and granuloma annulare. annular, Keratosis pilaris, Xeroderma, Panniculitis, Pyoderma gangrenosum, Acne, Rosacea, Lupus erythematosus, Pemphigus, Diaper dermatitis, Pityriasis rosea, Alopecia areata, Androgenic alopecia The cosmetic composition according to claim 15, wherein the skin ulcer is at least one selected from the group consisting of eczema, vitiligo, and decubitus ulcer.

17. The cosmetic composition according to claim 9, which is used for moisturizing the skin, restoring the skin barrier function, or preventing skin aging.

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