Composition for preventing or treating skin diseases comprising hydrogel

Abstract

The present invention relates to a composition or hydrogel for preventing or treating skin diseases. A composition or hydrogel according to one aspect exhibits an excellent therapeutic effect on various skin diseases characterized by skin barrier damage, excessive immune response, pruritus, and erythema. Accordingly, the present invention can be applied to not only systemic skin diseases but also localised skin diseases, and provides rapid recovery of skin lesions and alleviation of clinical symptoms by simultaneously inducing inflammation inhibition and tissue regeneration.

Description

A composition for the prevention or treatment of skin diseases comprising a hydrogel

[0001] The present invention relates to a composition for the prevention or treatment of skin diseases comprising a hydrogel.

[0002] Skin diseases are a group of conditions encompassing not only acute or chronic inflammatory responses occurring in the skin and skin appendages but also pathological conditions in which systemic immune and inflammatory abnormalities manifest on the skin, resulting in pruritus, erythema, edema, pain, a sensation of heat, skin barrier dysfunction, and various forms of tissue damage. These skin diseases include not only localized skin inflammation but also various conditions in which systemic inflammation or autoimmune reactions induce skin lesions.

[0003] Currently, various treatments are used for skin diseases, including topical corticosteroids, topical immunomodulators, antihistamines, and biologics.

[0004] However, topical formulations have structural limitations, such as insufficient adhesion and persistence, local side effects like skin atrophy and pigmentation changes, and issues with shedding in exudative or creatic areas. Furthermore, systemic administration-based drugs struggle to stably maintain high drug concentrations at the local lesion site, and safety concerns arise due to systemic side effects such as gastrointestinal disorders, cardiovascular abnormalities, nephrotoxicity, and an increased risk of infection with long-term use. In other words, existing treatments for skin diseases commonly share limitations in adequately addressing the pathophysiology of acute, chronic, and systemic skin diseases.

[0005] Therefore, there is a need to develop new treatment strategies for acute, chronic, and systemic skin diseases that can stably adhere to and remain at the lesion site, simultaneously induce inflammation suppression and tissue recovery, and minimize local and systemic side effects.

[0006] Accordingly, the inventors applied a hydrogel composition containing fibrinogen, laminin, and hyaluronic acid to various acute, chronic, and systemic skin inflammatory lesions and confirmed significant effects of alleviating skin inflammation, promoting tissue recovery, and improving lesions, and based on this, completed the present invention.

[0007] One aspect provides a pharmaceutical composition for the prevention or treatment of skin disease comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof as active ingredients.

[0008] Another aspect provides a pharmaceutical composition for the prevention or treatment of skin diseases comprising, as an active ingredient, a hydrogel or hydrogel patch comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof.

[0009] Another aspect provides a topical skin composition for the prevention or treatment of skin diseases comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof as active ingredients.

[0010] Another aspect provides a topical skin composition for the prevention or treatment of skin diseases comprising, as an active ingredient, a hydrogel or hydrogel patch comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof.

[0011] Another aspect provides a method for preventing or treating a skin disease comprising the step of administering a hydrogel or hydrogel patch containing fibrin or fibrin and fibrinogen; laminin or a laminin-derived peptide or protein; and hyaluronic acid or a salt thereof to an individual in need thereof.

[0012] Another aspect is to provide a hydrogel or hydrogel patch comprising fibrin or fibrin and fibrinogen; laminin or laminin-derived peptide or protein; and hyaluronic acid or its salt for use in the manufacture of medicines for the prevention or treatment of skin diseases.

[0013] Another aspect is to provide a hydrogel or hydrogel patch for the treatment or prevention of skin diseases comprising fibrin or fibrin and fibrinogen; laminin or laminin-derived peptide or protein; and hyaluronic acid or its salt.

[0014] One aspect provides a pharmaceutical composition for the prevention or treatment of skin diseases comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof as active ingredients.

[0015] Another aspect provides a composition for the prevention or treatment of skin diseases comprising, as an active ingredient, a hydrogel or hydrogel patch comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof.

[0016] Another aspect provides a topical skin composition for the prevention or treatment of skin diseases comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof as active ingredients.

[0017] Another aspect provides a topical skin composition for the prevention or treatment of skin diseases comprising, as an active ingredient, a hydrogel or hydrogel patch comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof.

[0018] Another aspect provides a quasi-drug composition for the prevention or treatment of skin diseases comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof as active ingredients.

[0019] Another aspect provides a quasi-drug composition for the prevention or treatment of skin diseases comprising, as an active ingredient, a hydrogel or hydrogel patch comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof.

[0020] Another aspect provides a method for preventing or treating a skin disease comprising the step of administering the above-mentioned hydrogel, hydrogel patch, or composition to an individual.

[0021] Another aspect provides the use of the above hydrogel, hydrogel patch, or composition in the manufacture of pharmaceutical preparations for the prevention or treatment of skin diseases.

[0022] Another aspect provides the use of the hydrogel, hydrogel patch, or composition for preventing or treating skin diseases.

[0023] The terms of the present specification, hydrogel, hydrogel patch, or composition may comprise fibrin and / or fibrinogen; laminin or laminin-derived peptide or protein; and / or hyaluronic acid or its salt. For example, the hydrogel, hydrogel patch, or composition may comprise fibrin or fibrinogen; laminin or laminin-derived peptide or protein; and hyaluronic acid or its salt.

[0024] The term “skin disease” in this specification refers to an acute, chronic, or systemic skin disease characterized by inflammation occurring in skin tissue as the main lesion, and specifically includes various inflammatory skin diseases such as atopic dermatitis, contact dermatitis, seborrheic dermatitis, psoriasis, eczema, urticaria, photodermatitis, irritant dermatitis, lichen simplex chronicus, dermatographism, impetigo, folliculitis, rosacea, and otitis externa.

[0025] In one embodiment, the skin disease is dermatitis, atopic dermatitis, contact dermatitis, seborrheic dermatitis, dermatitis herpetiformis, neurodermatitis, dermatomyositis, eczema, psoriasis-related dermatitis, psoriasis, urticaria, drug eruption, photodermatitis, irritant dermatitis, lichen simplex chronicus, dermatographism, folliculitis, impetigo, otitis externa, cutaneous vasculitis. It may be one or more diseases selected from the group consisting of vasculitis, cellulitis, erysipelas, dermatophytosis, acne vulgaris, and rosacea, but is not limited thereto.

[0026] In this specification, the term "pharmaceutical effective dose" means any amount of a composition used in the course of practicing the invention provided herein that is sufficient for the alleviation, inhibition of progression, prevention, or treatment of a disease, disorder, or condition, or one or more of its symptoms. The level of said effective dose may be determined by factors including the patient's health condition, the type and severity of the disease, sensitivity to the drug, the method of administration, the route of administration, and other factors well known in the medical field.

[0027] The terms “administering,” “applying,” “introducing,” “applying,” and “implanting” are used interchangeably and may mean the placement of a patch or composition according to one embodiment into an object by a method or route that results in at least partial localization of the patch or composition according to one embodiment to a desired site.

[0028] In this specification, the term "patch" may refer to a means having a certain shape that can be applied, attached, or contacted to a target area.

[0029] In one embodiment, the hydrogel, hydrogel patch, or composition may have properties intermediate between solid and liquid. The hydrogel, hydrogel patch, or composition may be amorphous, spherical, hemispherical, disc-shaped, or cylindrical. Additionally, for example, the diameter of the hydrogel patch may be 0.05 mm to 10 cm, 0.1 mm to 5 cm, 0.1 mm to 3 cm, or 0.2 mm to 1.5 cm, and may be provided in such a size or shape. Additionally, the hydrogel patch may be applied, implanted, attached, or in contact with a target site (e.g., a site of tissue damage) and deformed to conform to the shape of the damaged site.

[0030] In another embodiment, the hydrogel, hydrogel patch, or composition may be solid (including powder), semi-solid, or liquid. Additionally, for example, the hydrogel, hydrogel patch, or composition may undergo a reversible phase transition (e.g., depending on a change in temperature) to a solid (including powder), semi-solid, or liquid state. Since the hydrogel patch may undergo a reversible phase transition depending on ambient conditions such as temperature conditions, the hydrogel patch according to one embodiment may be produced and provided in a solid (including powder) or liquid state, and then converted into a hydrogel patch before, during, or after administration to a target site for use. For example, the hydrogel patch may be provided in a sol state containing fibrinogen, laminin, and hyaluronic acid, and the hydrogel patch according to one embodiment may be manufactured and used by using a substance (e.g., thrombin) that allows the user to convert fibrinogen into fibrin. Accordingly, a hydrogel patch according to one embodiment may be provided in the form of a prodrug, for example, a solid (powder), liquid (sol), or semi-solid composition comprising fibrin and / or fibrinogen; laminin; and / or hyaluronic acid. A composition provided in the form of a prodrug may be manufactured or modified into a hydrogel patch in vivo to function. Additionally, a hydrogel patch according to one embodiment may further comprise thrombin, or thrombin may be provided together as a kit.

[0031] In another embodiment, the hydrogel, hydrogel patch, or composition may be porous. Specifically, the surface of the hydrogel patch may have porosity (micropores). Without being limited to any specific theory, a hydrogel patch according to one embodiment may enhance the interaction between active substances by having porosity.

[0032] In one embodiment, the hydrogel, hydrogel patch, or composition may comprise fibrin and / or fibrinogen. The final pharmacological substance acting in vivo may comprise fibrin, but fibrinogen may be used instead of fibrin in the form of a prodrug. Additionally, depending on the amount of substance that converts fibrin to fibrinogen, the hydrogel, hydrogel patch, or composition according to one embodiment may partially comprise fibrinogen or thrombin. Accordingly, the present specification may additionally provide a prodrug comprising fibrinogen; laminin; and / or hyaluronic acid. The above fibrin or fibrinogen may be included at a concentration of 0.1 to 50 mg / ml, 0.1 to 25 mg / ml, 0.5 to 25 mg / ml, 1 to 25 mg / ml, 1 to 20 mg / ml, 5 to 25 mg / ml, 0.5 to 15 mg / ml, 1 to 15 mg / ml, 3 to 15 mg / ml, 5 to 15 mg / ml, 7 to 12 mg / ml, 8 to 25 mg / ml, 12 to 25 mg / ml, 15 to 24 mg / ml, 18 to 24 mg / ml, or 18 to 22 mg / ml.

[0033] The above fibrinogen glycoprotein is a hexamer composed of soluble α, β, and γ subunits produced in hepatocytes of the liver. Fibrinogen reacts with the enzyme thrombin to undergo a phase transition from soluble to insoluble fibrin polymer fibers.

[0034] The above-mentioned thrombin enzyme is a serine protease that transforms soluble fibrinogen into insoluble fibrin. The above-mentioned thrombin can play a role in gelling a hydrogel in a sol state by converting fibrinogen into fibrin.

[0035] In this specification, the term "laminin" may refer to an extracellular matrix protein constituting the basal lamina, specifically a heterotrimeric protein composed of α, β, and γ subunits. Accordingly, the laminin may include not only the full-length laminin protein but also laminin-derived peptides or proteins. For example, the laminin may be laminin-1, laminin-2, laminin-3, laminin-4, laminin-5A, laminin-5B, laminin-6, laminin-7, laminin-8, laminin-9, laminin-10, laminin-11, laminin-12, laminin-14, or laminin-15. Additionally, the laminin-derived peptide may be an α chain, a γ chain, or a β chain. The laminin may be included at a concentration of 1 to 100 μg / ml, 2 to 80 μg / ml, 5 to 50 μg / ml, 5 to 25 μg / ml, 8 to 20 μg / ml, 8 to 15 μg / ml, 10 to 70 μg / ml, 20 to 50 μg / ml, or 20 to 40 μg / ml.

[0036] In this specification, the term "hyaluronic acid" refers to a natural polymer found in various tissues, such as joint fluid, cartilage, connective tissue, and skin, as one of the representative glycosaminoglycans present in the body.

[0037] The above hyaluronic acid is a disaccharide polysaccharide composed of glycosidic bonds having β-(1→4) and β-(1→3) changes between D-glucuronic acid and N-acetyl-D-glucosamine, and has various molecular weights depending on the length of the disaccharide bonds. In one embodiment, the molecular weight of the hyaluronic acid may be 5,000 to 20,000,000 Da. More specifically, the molecular weight of the hyaluronic acid is 0.5 to 4.0 x 10⁻⁶. 6Da, 1.0 to 2.0 x 10 6 Da, or 1.5 to 1.8 x 10⁻⁶ 6 Da may be present. The hyaluronic acid may be included at a concentration of 10 μg / ml to 10 mg / ml, 10 μg / ml to 5 mg / ml, 50 μg / ml to 5 mg / ml, 100 μg / ml to 3 mg / ml, 200 μg / ml to 1 mg / ml, 500 μg / ml to 5 mg / ml, 500 μg / ml to 8 mg / ml, 500 μg / ml to 4 mg / ml, 500 μg / ml to 3 mg / ml, 300 μg / ml to 3 mg / ml, 1 mg / ml to 8 mg / ml, 1 mg / ml to 5 mg / ml, or 1 mg / ml to 3 mg / ml.

[0038] In other embodiments, the hydrogel, hydrogel patch, or composition may further include a cell growth factor. The cell growth factor may be a neuronal growth factor, a vascular endothelial growth factor, a fibroblast growth factor, a bone morphogenetic protein (BMP), an epidermal growth factor (EGF), a hepatocyte growth factor (HGF), a transforming growth factor (TGF), or a combination thereof, but is not limited thereto.

[0039] In one embodiment, the hydrogel, hydrogel patch, or composition comprises neuronal growth factor, vascular endothelial growth factor, fibroblast growth factor (FGF), bone morphogenetic protein (BMP), epidermal growth factor (EGF), hepatocyte growth factor (HGF), transforming growth factor (TGF), placental growth factor (PIGF), macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), neuropilin, FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, erythropoietin (EPO), BMP-2, BMP-4, BMP-7, BMP-9, TGF-β, IGF-1, osteopontin, pleiotrophin, activin, endothelin-1, It may include, but is not limited to, BDNF, GDNF, CNTF, cAMP, NT, NT-3, NT-4, T3, SHH, PDGF, VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, or combinations thereof.

[0040] The concentration of the cell growth factor included in the hydrogel, hydrogel patch, or composition may vary depending on the type of growth factor, but generally, it may be included at a concentration of 1 ng / ml to 1,000 ng / ml or 0.1 μM to 100 μM. The cell growth factor may play a role in enhancing the tissue damage recovery effect of a hydrogel patch or composition containing fibrin, laminin, and hyaluronic acid.

[0041] In other embodiments, the hydrogel, hydrogel patch, or composition may include or substantially not include collagen. Without being limited to any particular theory, the collagen may be included as a component of the composition or may not be substantially included, but in one aspect, not including it may be more advantageous than including it.

[0042] Additionally, the hydrogel, hydrogel patch, or composition may not substantially comprise cells. In this specification, "substantially does not comprise" means that collagen or cells are included to an extent that they do not affect the activity or pharmacological activity of the hydrogel, hydrogel patch, or composition, or are not included at all. The hydrogel, hydrogel patch, or composition according to one embodiment may be distinguished from cell therapies generally used for the regeneration of damaged tissue by not substantially comprising cells.

[0043] In one embodiment, the hydrogel, hydrogel patch, or composition may be composed of fibrin and / or fibrinogen; laminin or a laminin-derived peptide; and hyaluronic acid or a salt thereof. Additionally, one or more of the components mentioned herein may be additionally included. For example, the hydrogel, hydrogel patch, or composition may be composed of fibrin and / or fibrinogen; laminin or a laminin-derived peptide; hyaluronic acid or a salt thereof; optionally thrombin; optionally collagen; and optionally a cell growth factor.

[0044] The dosage of the above composition according to one embodiment may be 0.01 mg to 10,000 mg, 0.1 mg to 1,000 mg, 1 mg to 100 mg, 0.01 mg to 1,000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg. However, the dosage may be prescribed differently depending on factors such as the formulation method, administration method, patient's age, weight, gender, pathological condition, food, time of administration, route of administration, excretion rate, and response sensitivity, and a person skilled in the art may appropriately adjust the dosage by considering these factors. The number of administrations may be one or two or more within the range of clinically acceptable side effects, and regarding the administration site, it may be administered at one or two or more sites. For animals other than humans, the above dosage may be administered at the same dose per kg as for humans, or calculated based on the ratio of organ volumes (e.g., heart, etc.) between the target animal and humans (e.g., average value). Possible routes of administration may include oral, sublingual, parenteral (e.g., subcutaneous, intramuscular, intra-arterial, intraperitoneal, intradural, or intravenous), rectal, local (including transdermal), inhalation, and injection, or insertion of an implantable device or substance. Examples of animals targeted for treatment according to one embodiment include various animal species including humans, and specifically include mammals such as humans, monkeys, mice, rats, rabbits, sheep, cattle, dogs, horses, camels, pigs, sugar gliders, etc., as well as birds (avian species) such as parrots and reptilian species such as lizards.

[0045] A pharmaceutical composition according to one embodiment may include pharmaceutically acceptable carriers and / or additives. For example, it may include sterile water, physiological saline, a common buffer (phosphoric acid, citric acid, other organic acids, etc.), a stabilizer, a salt, an antioxidant (ascorbic acid, etc.), a surfactant, a suspending agent, an isotonic agent, or a preservative. For topical administration, it may also include combinations with organic materials such as biopolymers, inorganic materials such as hydroxyapatite, specifically collagen matrices, polylactic acid polymers or copolymers, polyethylene glycol polymers or copolymers, and chemical derivatives thereof.

[0046] A pharmaceutical composition according to one embodiment may appropriately include, if necessary depending on the method of administration or dosage form, a suspending agent, a solubilizing agent, a stabilizer, an isotonic agent, a preservative, an anti-adsorption agent, a surfactant, a diluent, an excipient, a pH adjuster, an analgesic agent, a buffer, a reducing agent, an antioxidant, etc. Pharmaceutically acceptable carriers and formulations suitable for the present invention, including those exemplified above, are described in detail in the literature [Remington's Pharmaceutical Sciences, 19th ed., 1995]. A pharmaceutical composition according to one embodiment may be prepared in a unit dose form or contained in a multi-dose container by formulating using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily carried out by a person skilled in the art to which the invention pertains. In this case, the dosage form may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or in the form of a powder, granule, tablet, or capsule.

[0047] The above composition may be formulated into an oral or parenteral administration formulation. The oral administration formulation may be a granule, powder, liquid, tablet, capsule, dry syrup, or a combination thereof. The parenteral administration formulation may be an injection or a topical application.

[0048] The term "topical preparation for the skin" in this specification refers to a formulation intended for local application by applying directly to or attaching to the skin. The topical preparation for the skin may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, drug-containing bandage, lotion, or a combination thereof. The topical preparation for the skin may be appropriately formulated as needed with ingredients commonly used in topical preparations for the skin, such as cosmetics, pharmaceuticals, or quasi-pharmaceuticals, for example, aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, UV absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or combinations thereof. The above topical preparation may also appropriately incorporate metal chelating agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid; caffeine, tannin, bellapamil, licorice extract, glablidin, hot water extract of the fruit of *calin*; various herbal medicines; preparations such as tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts; and sugars such as vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, kojic acid, glucose, fructose, and trehalose.

[0049] In addition, the above composition may be a quasi-drug composition. The term "quasi-drug" refers to an article that falls under one of the following categories: fibers, rubber products, or similar items used for the purpose of treating, alleviating, managing, or preventing diseases in humans or animals; items that have a weak effect on the human body or do not act directly on the human body and are not instruments or machines, or similar items; and preparations used for sterilization, insecticidal, and similar purposes for the prevention of infection. It excludes articles used for the purpose of diagnosing, treating, alleviating, managing, or preventing diseases in humans or animals that are not instruments, machines, or devices, and articles used for the purpose of exerting pharmacological effects on the structure and function of humans or animals that are not instruments, machines, or devices. It may also include external skin preparations and personal hygiene products. When the above hydrogel, hydrogel, hydrogel patch, or composition is added to a quasi-drug composition for the purpose of preventing or improving skin diseases, the above hydrogel, hydrogel, hydrogel patch, or composition may be added as is or used together with other quasi-drug ingredients, and may be used appropriately according to conventional methods. The mixture of active ingredients can be appropriately determined according to the purpose of use (prevention, health, or therapeutic treatment).

[0050] Another aspect provides a method for preparing a hydrogel patch by adding thrombin to a composition in a sol state comprising fibrinogen; laminin or a laminin-derived peptide; and hyaluronic acid or a pharmaceutically acceptable salt thereof.

[0051] The above method may also further include the step of incorporating a cell growth factor into the composition in the sol state.

[0052] The above method may also include a step of gelling by adding thrombin, and then gelling by adding thrombin again. The gelling may be performed at 10 to 40°C for 5 minutes to 3 hours. The hydrogel patch may be produced in various shapes or sizes depending on the shape of the mold.

[0053] The above method may also include the step of low-temperature preservation or cryopreservation of the hydrogel patch in a solution at 4 to -210°C. The solution may include DMSO (Dimethyl sulfoxide), but any solution that does not substantially alter the chemical or physical properties of the hydrogel patch may be used. The form or activity of the hydrogel patch does not substantially change even when low-temperature preservation or cryopreservation.

[0054] The above hydrogel patch, fibrin and / or fibrinogen, laminin, hyaluronic acid, or cell growth factor are as described above.

[0055] Another aspect provides a method for low-temperature preservation or cryopreservation of the above hydrogel patch in a solution (e.g., DMSO) at 4 to -210°C.

[0056] A composition or hydrogel according to one aspect exhibits excellent therapeutic effects on various skin diseases characterized by skin barrier damage, excessive immune response, itching, and erythema. Accordingly, it is applicable to systemic skin diseases as well as local skin diseases in general, and provides rapid recovery of skin lesions and alleviation of clinical symptoms by simultaneously inducing inflammation suppression and tissue regeneration.

[0057] Figure 1 is a photograph showing changes in lesions before and 2 days after application of a hydrogel patch according to one embodiment in a clinical animal with induced atopic dermatitis.

[0058] Figure 2 is a photograph showing changes in lesions before and 3 weeks after the application of a hydrogel patch according to one embodiment in a clinical animal with skin lesions accompanied by chronic inflammation and deep tissue necrosis.

[0059] Figure 3 shows photographs of the external auditory canal lesions of a clinical animal with induced chronic otitis externa before and one week after the application of a hydrogel patch.

[0060] The present invention will be explained in detail below through reference examples and embodiments. However, the following reference examples and embodiments are merely illustrative of the present invention and should not be construed as limiting the present invention.

[0061]

[0062] Example 1. Preparation of a hydrogel patch

[0063] 1.1. Hydrogel Formulation

[0064] Fibrinogen was dissolved in physiological saline or PBS at a sol state at 20 mg / ml, hyaluronic acid was dissolved in physiological saline or PBS at 5 mg / ml, and thrombin was dissolved in physiological saline or PBS at 200 U / ml. Subsequently, sol-state hydrogels were prepared by combining sol-state fibrinogen at 1 mg / ml, 5 mg / mL, or 10 mg / ml; laminin at 5 µg / ml, 10 µg / ml, 30 µg / ml, or 50 µg / ml; and hyaluronic acid at 0.1 mg / ml, 0.5 mg / ml, 1.0 mg / ml, or 1.5 mg / ml.

[0065] 1.2. Manufacture of Hydrogel Patch

[0066] Thrombin in a sol state was mixed with fibrinogen, laminin, and hyaluronic acid prepared in Example 1.1, and then dispensed into a sterile Parafilm or petri dish. Subsequently, a hydrogel patch was prepared by curing at room temperature or 37°C through a sol-gel phase transition.

[0067]

[0068] Experimental Example 1. Confirmation of the therapeutic effect of a hydrogel patch on atopic dermatitis

[0069] The therapeutic effect of a hydrogel patch on atopic dermatitis was confirmed in clinical animals suffering from atopic dermatitis.

[0070] Specifically, the subject small dogs exhibited typical atopic dermatitis lesions, including extensive erythema, erosion, exudation, crust, and hair loss, on the lateral flank and dorsal region.

[0071] A hydrogel patch (410 µg / kg) of 200 µl to 10 ml prepared by the method of Example 1 was cut to fit the size of the lesion and attached to the subject small dog in close contact to cover the entire surface of the wound. The improvement of the lesion was evaluated for 2 days after administration of the hydrogel patch, and the results are shown in Table 1 and Figure 1.

[0072] Table 1 summarizes the degree of lesion improvement by evaluation item 2 days after the application of a hydrogel patch to clinical animals with induced atopic dermatitis, and Figure 1 is a photograph showing the changes in lesions before and after the application of a hydrogel patch to clinical animals with induced atopic dermatitis.

[0073] Evaluation Criteria Results Observed 2 Days After Hydrogel Patch Application Reduction in Erythema Significant reduction in extensive redness Suppression of Inflammation Reduction in skin surface heat sensation and inflammatory edema Disappearance of Exudate and Crusts Most exudate and thick crusts removed Re-epithelialization Progression Clear recovery of damaged epidermal layer Reduction in Lesion Size The lesion area was visibly reduced Normalization of Epidermal Thickness Skin wrinkles and tone restored to near-normal state Reduction in Scratches Secondary excoriation almost completely disappeared

[0074] As shown in Table 1 and Figure 1, when the hydrogel patch of the present invention was applied, erythema in the lesion area was significantly reduced, and inflammatory redness and edema were also clearly alleviated. In addition, most of the previously formed exudate and thick crusts were removed, and the exposed eroded areas rapidly re-epithelialized to restore a smooth surface. The total area of ​​the lesion also decreased, blurring the boundary with the surrounding normal skin, and the thickness of the epidermis and skin tone were restored to a level similar to normal skin. Furthermore, the stabilization of the overall skin condition was confirmed, with the almost complete disappearance of excoriation caused by persistent itching due to atopy.

[0075] Through these results, it was confirmed that the hydrogel according to one embodiment of the present invention has therapeutic effects such as inflammation inhibition, wound protection, promotion of re-epithelialization, and reduction of lesions in skin damaged by atopic dermatitis.

[0076]

[0077] Experimental Example 2. Confirmation of the therapeutic effect of a hydrogel patch on chronic inflammation and tissue necrosis

[0078] The chronic inflammation improvement and tissue regeneration effects of the hydrogel according to one embodiment of the present invention were evaluated on clinical animals (rabbits) suffering from skin lesions accompanied by chronic inflammation and deep tissue necrosis.

[0079] Specifically, the study targeted lesions in small dogs in which chronic inflammation and tissue necrosis had not fully recovered despite repeated treatment. The subjects exhibited characteristics such as exposure of the subcutaneous tissue and part of the fascial layer due to severe skin defects, extensive presence of necrotic tissue mixed with red and grayish-white colors, bloody or exudate or pus observed throughout the wound surface, and accompanying skin thickening and edema caused by chronic inflammation.

[0080] A hydrogel patch of 200 µl to 5 ml prepared by the method of Example 1 was cut to fit the size of the lesion on the subject animal, adhered to cover the entire surface of the wound, and then the wound was sutured. The degree of tissue regeneration and improvement of chronic inflammation was evaluated by item for a total of 3 weeks after the application of the hydrogel patch, and the results are shown in Table 2 and Figure 2.

[0081] Table 2 summarizes the degree of wound healing and lesion improvement by evaluation item after applying a hydrogel patch to clinical animals with induced chronic inflammation and deep tissue necrosis, and Figure 2 is a photograph showing the changes in lesions before and after the application of a hydrogel patch to clinical animals with induced skin lesions accompanied by chronic inflammation and deep tissue necrosis.

[0082] Evaluation Criteria Before Hydrogel Patch Application 3 Weeks After Hydrogel Patch Application Skin Defect Exposure of subcutaneous tissue and fascia layer, large defect area Most defect area regenerated, skin surface restored Necrotic Tissue Presence of extensive necrotic tissue mixed with red and grayish-white Disappearance of necrotic tissue, healthy granulation tissue and epidermal remodeling Exudate Large amount of hemorrhagic or purulent exudate Most exudate disappeared, dry and stable wound Skin Thickening / Edema Distinct thickening and edema due to chronic inflammation Reduced edema, skin thickness returned to normal levels

[0083] As shown in Table 2 and Figure 2, extensive tissue necrosis in the lesion area was inhibited after the application of the hydrogel patch, and distinct tissue regeneration and epithelialization proceeded throughout the wound. After 3 weeks of treatment, the open wound was almost completely contracted, and the therapeutic effect of the hydrogel was confirmed by observing that healthy new tissue uniformly covered the wound area. These results indicate that the hydrogel patch according to one embodiment of the present invention can be usefully utilized as a composition for treating chronic wounds, tissue necrosis, and intractable inflammatory skin lesions.

[0084]

[0085] Experimental Example 3. Confirmation of the therapeutic effect of a hydrogel patch on otitis externa

[0086] The effect of a hydrogel patch according to one embodiment of the present invention on improving inflammation and clinical symptoms was evaluated in small dogs with chronic otitis externa.

[0087] Specifically, the study was conducted on small dogs diagnosed with chronic bilateral otitis externa, characterized by chronic redness and thickening of the external auditory canal, accompanied by yellow purulent discharge and a foul odor. Before the hydrogel patch treatment, the skin of the external auditory canal of the subject small dogs showed severe erythema and edema, a large amount of viscous purulent discharge had accumulated in the lumen of the external auditory canal, and a distinct foul odor was observed around the external auditory canal.

[0088] For the external ear lesions of the small dog subject to the above, 200 µl to 5 ml of the hydrogel patch of Example 1, cut to fit the shape of the entrance of the external auditory canal and the lesion, was applied and attached so as to adhere closely along the inner wall of the external auditory canal.

[0089] One week after the application of the hydrogel patch, the degree of improvement in otitis externa was evaluated by item, such as erythema, edema, discharge, odor, and the condition of the skin of the external auditory canal, and the results are shown in Table 3 and Figure 3.

[0090] Table 3 compares the otitis externa lesions in clinical animals with induced chronic otitis externa before and one week after the application of a hydrogel patch for each evaluation item, and Figure 3 shows photographs of the otitis externa lesions in clinical animals with induced chronic otitis externa before and one week after the application of a hydrogel patch.

[0091] Evaluation Criteria Before Hydrogel Patch Application 1 Week After Hydrogel Patch Application External Auditory Canal Erythema Severe redness and purple lesions throughout the external auditory canal Erythema significantly decreased and skin tone lightened External Auditory Canal Edema and Thickening Narrowing of the lumen due to thickening and edema of the external auditory canal walls Lumen opened as edema and thickening decreased Discharge (Exudate / Discharge) Large accumulation of yellow purulent discharge Discharge significantly decreased or barely observed Odor Strong odor emanating from the external ear area Bilateral posterior inflammation and odor distinctly disappeared or significantly decreased External Auditory Canal Skin Condition Uneven surface and prominent inflammatory changes Skin surface became smooth and signs of chronic inflammation alleviated

[0092] As shown in Table 3 and Figure 3, after the application of the hydrogel patch, purulent secretions in the external auditory canal were almost completely eliminated, severe erythema and edema were significantly reduced, and the thickening of the skin of the external auditory canal was alleviated, restoring the lumen to a more open form. These results indicate that the hydrogel patch of the present invention is effective in improving or treating otitis externa by effectively alleviating inflammatory lesions in an animal model of chronic otitis externa, reducing secretions and odors, and promoting the recovery of damaged skin of the external auditory canal.

Claims

1. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and A pharmaceutical composition for the prevention or treatment of skin disease, comprising hyaluronic acid or a salt thereof as an active ingredient.

2. In Claim 1, A pharmaceutical composition, wherein the above composition is a hydrogel or a hydrogel patch.

3. In Claim 1, A pharmaceutical composition comprising the above fibrin or fibrinogen at a concentration of 0.1 to 50 mg / ml, the above laminin at a concentration of 1 to 100 μg / ml, or the above hyaluronic acid at a concentration of 10 μg / ml to 10 mg / ml.

4. In Claim 1, A pharmaceutical composition that additionally comprises thrombin.

5. In Claim 1, A pharmaceutical composition further comprising a cell growth factor.

6. In Claim 5, The above cell growth factors are neuronal growth factor, vascular endothelial growth factor, fibroblast growth factor (FGF), bone morphogenetic protein (BMP), epidermal growth factor (EGF), hepatocyte growth factor (HGF), transforming growth factor (TGF), placental growth factor (PIGF), macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), neuropilin, FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, erythropoietin (EPO), BMP-2, BMP-4, BMP-7, BMP-9, TGF-β, IGF-1, osteopontin, pleiotrophin, activin, endothelin-1, BDNF, GDNF, CNTF, cAMP, A pharmaceutical composition that is NT, NT-3, NT-4, T3, SHH, PDGF, VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, or a combination thereof.

7. In Claim 2, A pharmaceutical composition in which the above hydrogel or hydrogel patch does not contain cells.

8. In Claim 2, A pharmaceutical composition in which the above hydrogel or hydrogel patch has porosity on its surface.

9. In Claim 2, A pharmaceutical composition in which the above hydrogel or hydrogel patch undergoes a reversible phase transition to a solid, semi-solid, or liquid state depending on temperature.

10. In Claim 1, The above skin diseases include dermatitis, atopic dermatitis, contact dermatitis, seborrheic dermatitis, dermatitis herpetiformis, neurodermatitis, dermatomyositis, eczema, psoriasis-related dermatitis, psoriasis, urticaria, drug eruption, photodermatitis, irritant dermatitis, lichen simplex chronicus, dermatographism, folliculitis, impetigo, otitis externa, and cutaneous vasculitis. A pharmaceutical composition comprising one or more diseases selected from the group consisting of cellulitis, erysipelas, dermatophytosis, acne vulgaris, and rosacea.

11. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and A topical skin composition for the prevention or treatment of skin diseases, comprising hyaluronic acid or a salt thereof as an active ingredient.

12. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and A method for preventing or treating a skin disease comprising the step of administering a hydrogel or hydrogel patch containing hyaluronic acid or a salt thereof as an active ingredient to an individual in need thereof.

13. For use in the manufacture of medicines for the prevention or treatment of skin diseases. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and Use of a hydrogel or hydrogel patch containing hyaluronic acid or its salt as an active ingredient.

14. For the treatment or prevention of skin diseases, Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and Use of a hydrogel or hydrogel patch containing hyaluronic acid or its salt as an active ingredient.

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