Composition for preventing or treating inflammatory diseases comprising hydrogel

Abstract

The present invention relates to a composition or hydrogel for preventing or treating inflammatory diseases. The composition or hydrogel according to one aspect exhibits a clear improvement effect when applied topically to various inflammatory lesions such as inflammation of the intestinal mucosa, chronic diarrhea, redness, swelling, and bleeding of periodontal tissue, and alveolar bone damage, thus providing a useful means for preventing or treating inflammatory diseases.

Description

Composition for the prevention or treatment of inflammatory diseases comprising a hydrogel

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

[0002] Inflammatory diseases are a group of conditions characterized by persistent abnormal or excessive inflammatory responses in various bodily structures, such as the mucosa, soft tissues, and supporting tissues, which cause pain, swelling, tissue damage, and functional impairment. Among these, inflammatory bowel disease (IBD) and periodontal disease are representative inflammatory diseases occurring in the intestinal tract and oral and periodontal tissues; they are recognized as significant medical and social issues due to their recurring nature and the need for long-term management.

[0003] Currently, 5-aminosalicylic acid (5-ASA) preparations, steroids, immunomodulators, and biological agents (anti-TNF, anti-IL-12 / 23, etc.) are used to treat inflammatory bowel disease (IBD), but various systemic side effects, such as increased risk of infection, hepatotoxicity, bone marrow suppression, and metabolic abnormalities, have been reported with long-term use. In addition, there is a limitation in that it is difficult for the drugs to directly reach and maintain high concentrations in intestinal lesions.

[0004] Meanwhile, the treatment of periodontal disease utilizes mechanical procedures such as scaling and root planning, antibiotic administration, anti-inflammatory agents, or topical oral agents (gels, mouthwashes, etc.); however, topical agents often have poor retention due to mucosal and salivary secretion and frequently fail to provide sufficient therapeutic effects for deep lesions such as alveolar bone defects or abscesses. Furthermore, conventional antibiotic-based treatments are accompanied by issues of resistance, tissue toxicity, and difficulties with long-term use.

[0005] Accordingly, the inventors devised a hydrogel composition comprising fibrinogen, laminin, and hyaluronic acid to solve these problems, and completed the present invention by confirming that the composition exhibits excellent anti-inflammatory effects and tissue recovery-promoting effects in various inflammatory lesions such as intestinal mucosal inflammation, chronic diarrhea, alveolar bone inflammation, and periodontal tissue damage.

[0006] One aspect provides a pharmaceutical composition for the prevention or treatment of inflammatory diseases comprising, as active ingredients, fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof.

[0007] Another aspect provides a pharmaceutical composition for the prevention or treatment of inflammatory 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.

[0008] Another aspect provides a topical skin composition for the prevention or treatment of inflammatory 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.

[0009] Another aspect provides a topical skin composition for the prevention or treatment of inflammatory 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.

[0010] Another aspect provides a method for preventing or treating an inflammatory 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.

[0011] 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 inflammatory diseases.

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

[0013] One aspect provides a pharmaceutical composition for the prevention or treatment of inflammatory 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.

[0014] Another aspect provides a composition for the prevention or treatment of inflammatory 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.

[0015] Another aspect provides a topical skin composition for the prevention or treatment of inflammatory 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.

[0016] Another aspect provides a topical skin composition for the prevention or treatment of inflammatory 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.

[0017] Another aspect provides a quasi-drug composition for the prevention or treatment of inflammatory 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.

[0018] Another aspect provides a quasi-drug composition for the prevention or treatment of inflammatory 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.

[0019] Another aspect provides a method for preventing or treating an inflammatory disease, comprising the step of administering the above-mentioned hydrogel, hydrogel patch, or composition to an individual.

[0020] 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 inflammatory diseases.

[0021] Another aspect provides the use of the hydrogel, hydrogel patch, or composition for preventing or treating inflammatory diseases.

[0022] 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.

[0023] The term “inflammatory disease” in this specification refers to a group of diseases characterized by acute or chronic inflammatory responses in tissues as the primary lesion, and includes, in particular, inflammatory pathological conditions occurring in the gastrointestinal tract or periodontal tissue.

[0024] The above inflammatory disease may be inflammatory bowel disease (IBD) or periodontal disease.

[0025] The term “inflammatory bowel disease (IBD)” in this specification refers to a group of diseases in which an acute or chronic inflammatory response occurs throughout the mucosa, submucosa, or entire intestinal wall of the intestinal tract, and may also include other intestinal diseases characterized by mucosal inflammation, persistent diarrhea, and inflammatory damage to the intestines. The inflammatory bowel disease may be accompanied by various gastrointestinal symptoms and histological changes, such as abdominal pain, diarrhea, bloody stools, mucus discharge, weight loss, damage to the intestinal wall, ulcer formation, and malabsorption.

[0026] In one embodiment, the inflammatory bowel may be one or more diseases selected from the group consisting of nonspecific chronic enteritis, indeterminate colitis, Crohn's disease, ulcerative colitis, microscopic colitis or subtypes thereof such as lymphocytic colitis, collagenous colitis, autoimmune enterocolitis, granulomatous enteritis, irritable bowel syndrome (IBS), or Behcet's colitis, but is not limited thereto.

[0027] The term “Periodontal disease” as used in this specification comprehensively refers to acute or chronic inflammatory lesions occurring in the periodontal tissues (gingiva, periodontal ligament, cementum, alveolar bone) and surrounding soft tissues, which are structures supporting the teeth. Such periodontal disease may exhibit clinical characteristics such as pain, redness, swelling, bleeding, exudate formation, tissue damage, alveolar bone resorption, mucosal rupture, or delayed epithelialization, and may also include infectious complications such as impaired healing after tooth extraction or abscess formation.

[0028] In one embodiment, the periodontal disease may be one or more diseases selected from the group including gingivitis, periodontitis, alveolar bone inflammation or alveolar bone defect, peri-implantitis, pulpitis, periapical abscess, stomatitis, mouth ulcers, and oral ulcers, but is not limited thereto.

[0029] 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.

[0030] 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.

[0031] 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.

[0032] 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.

[0033] 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.

[0034] 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.

[0035] 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.

[0036] 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.

[0037] 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.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 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-βIGF-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.

[0043] 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.

[0044] 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.

[0045] 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.

[0046] 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.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] 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.

[0051] The term “topical preparation for skin” in this specification refers to a formulation for local application by applying directly to or attaching to the skin. The topical preparation for 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 skin may be appropriately formulated as needed with ingredients commonly used in topical preparations for 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.

[0052] 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 topical 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 inflammatory 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).

[0053] The above-mentioned quasi-drugs may include products used for the purpose of alleviating local inflammatory reactions in the intestines or oral and periodontal tissues, protecting mucous membranes, promoting wound healing, or managing hygiene. Specifically, the above-mentioned quasi-drugs may include, but are not limited to, a perianal soothing patch for relieving perianal irritation caused by diarrhea or intestinal inflammation, a rectal protective or soothing gel for protecting the rectal or anal mucosa or relieving inflammation, a rectal suppository applicable to lesions of proctitis or local intestinal inflammation, a rectal enema formulation for protecting the mucosa or relieving inflammation, a mouth rinse for anti-inflammatory purposes, a gingival protective or soothing gel, a buccal protective film, an oral ulcer relief topical agent, a toothpaste or periodontal care toothpaste with anti-inflammatory and antibacterial functions, and an oral / gingival patch that helps restore periodontal tissue.

[0054] 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.

[0055] The above method may also further include the step of incorporating a cell growth factor into the composition in the sol state.

[0056] 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.

[0057] 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.

[0058] The above hydrogel patch, fibrin and / or fibrinogen, laminin, hyaluronic acid, or cell growth factor are as described above.

[0059] 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.

[0060] The terms, methods, effects, etc. described for the above inventions apply equally between each invention.

[0061] A composition or hydrogel according to one aspect not only effectively suppresses acute and chronic inflammatory responses, but also shows a distinct improvement effect when applied topically to various inflammatory lesions such as inflammation of the intestinal mucosa, chronic diarrhea, redness, swelling, bleeding of periodontal tissues, and damage to the alveolar bone, thereby providing a useful means for the prevention or treatment of inflammatory diseases.

[0062] Figure 1 is a photograph showing the condition of the tissue around the anus of a sugar glider before and one week after administration of hydrogel to clinical animals with chronic inflammatory bowel disease and chronic diarrhea caused by it.

[0063] Figure 2 is a photograph showing the condition of the gums and surrounding alveolar bone tissues before and after the application of hydrogel.

[0064] Figure 3 is a photograph showing the hydrogel administration process and the radiographic images and changes in the alveolar bone condition before and after administration in sequence.

[0065] 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.

[0066]

[0067] Example 1. Preparation of a hydrogel patch

[0068] 1.1. Hydrogel Formulation

[0069] 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.

[0070] 1.2. Manufacture of Hydrogel Patch

[0071] 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.

[0072] Experimental Example 1. Confirmation of the effect of a hydrogel patch in improving colitis and chronic diarrhea

[0073] The effects of the hydrogel patch of the present invention on alleviating intestinal inflammation and improving clinical symptoms were evaluated on sugar gliders exhibiting chronic inflammatory bowel condition and chronic diarrhea that had persisted for more than one year.

[0074] Specifically, the subject sugar glider did not show improvement in diarrhea symptoms despite continuing general veterinary treatment and standard care for more than one year, and severe contamination, hair loss, inflammation, and tissue damage around the anus occurred repeatedly. Abdominal radiography of the subject sugar glider prior to hydrogel administration also revealed findings suggestive of intestinal inflammation, and persistent lesions attributable to chronic diarrhea were identified, including inflammatory exudate, fecal residue, skin irritation, and hyperkeratosis in the anal area.

[0075] For the above-mentioned sugar gliders, 200 µl to 5 ml of hydrogel (410 µg / kg) prepared by the method of Example 1 was administered into the bodily cavity or rectum, and clinical symptoms including improvement in diarrhea, degree of recovery of perianal tissue, change in body weight, and degree of reduction in inflammation were monitored for one week after administration, and the results are presented in Table 1 and Figure 1.

[0076] Table 1 compares the changes in clinical symptoms by category before and one week after administration of hydrogel in clinical animals with chronic inflammatory bowel disease and chronic diarrhea caused by it, and Figure 1 is a photograph showing the condition of the tissue around the anus of sugar gliders before and one week after administration of hydrogel in clinical animals with chronic inflammatory bowel disease and chronic diarrhea caused by it.

[0077] Evaluation Criteria Before Hydrogel Administration 1 Week After Hydrogel Administration Diarrhea Chronic diarrhea lasting over 1 year, no improvement Complete resolution of diarrhea, normal stool formation Perianal Inflammation Severe redness, exudation, contamination, and foul odor Cleansing of the perianal area and reduction of inflammation Skin Damage Hair loss and sores due to repetitive irritation Resolution of sores and irritation, skin recovery Body Weight Weight loss due to chronic diarrhea Confirmation of weight gain

[0078] As shown in Table 1 and Figure 1, after the application of the hydrogel according to one embodiment of the present invention, severe skin inflammation and contamination caused by chronic diarrhea around the anus completely disappeared, and the skin surface was restored to a normal state. In addition, the feces normalized and persistent diarrhea symptoms were resolved, confirming the therapeutic effect of the hydrogel. These results suggest that the hydrogel according to one embodiment of the present invention is effective in treating inflammatory bowel diseases, including intestinal mucosal damage and chronic diarrhea.

[0079]

[0080] Experimental Example 2. Confirmation of the effect of the hydrogel patch in improving alveolar bone inflammation

[0081] 2.1 Confirmation of Effect on Improving Alveolar Bone Inflammation in Small Dogs

[0082] The effect of the hydrogel patch of the present invention on improving periodontitis and alveolar bone inflammation was evaluated in small dogs exhibiting moderate to severe periodontitis and alveolar bone inflammation.

[0083] Specifically, severe tartar buildup and gingivitis were observed in multiple teeth of the subject small dogs, and some teeth showed advanced alveolar bone resorption and inflammation to the extent that extraction was necessary. After extraction, abscesses, redness, necrotic tissue, and alveolar bone exposure were confirmed at the lesion sites.

[0084] For the subject small dogs mentioned above, after extracting the affected tooth, 100 µl to 5 ml of hydrogel (410 µg / kg) prepared by the method of Example 1 was directly filled and applied to the extraction site and the exposed alveolar bone immediately after extraction. The opposite extraction site was set as the comparison site for hydrogel therapeutic efficacy by performing only extraction and rinsing without applying hydrogel.

[0085] Clinical aspects, including the degree of inflammation reduction, alveolar bone recovery, mucosal tissue condition, and the presence of abscesses and exudates, were evaluated while observing the healing progress for 18 days after administration of the hydrogel, and the results are presented in Table 2 and Figure 2.

[0086] Table 2 compares the changes in clinical symptoms by category before and 18 days after application of hydrogel in clinical animals with induced alveolar bone inflammation and alveolar bone defects, and Figure 2 is a photograph showing the condition of the gums and surrounding tissues of the alveolar bone before and after application of hydrogel.

[0087] Evaluation Criteria Before Hydrogel Application 18 Days After Hydrogel Application Alveolar Bone Integrity Alveolar bone defect and resorption, presence of abscess and necrotic tissue Radiographic findings: Disappearance of defect space and alveolar bone regeneration Gingival Inflammation Severe redness, swelling, and bleeding Reduced redness and edema, mucosa-like Exudate and Abscess Large amount of abscess and exudate observed Disappearance of abscess and exudate Mucosa and Tissue Damage Severe wound, mucosal rupture, and irritation Progression of epithelialization, good mucosal recovery Overall Healing Persistent inflammation and malodor Disappearance of inflammation, tissue recovery and stabilization

[0088] As shown in Table 2 and Figure 2, the oral condition of the subject small dogs was evaluated before and 18 days after the application of the hydrogel. As a result, redness and swelling were significantly reduced in the experimental area where the hydrogel was applied, and the gingival margin was relatively smooth and showed a color tone similar to normal gingiva. In addition, it was observed that the mucosa and alveolar bone exposed areas of the extraction site were covered with uniform epithelialization and good granulation tissue formation. On the other hand, in the control area where the hydrogel was not applied, redness and swelling remained, and gingival bleeding and gingivitis were still present. These results indicate that the hydrogel composition of the present invention has the effect of alleviating inflammatory responses, promoting healing of the extraction site, and improving the regenerative environment of alveolar bone and gingival tissues when applied locally to periodontal and extraction lesions accompanied by alveolar bone inflammation.

[0089]

[0090] 2.2 Effect of Improving Alveolar Bone Inflammation in Guinea Pigs

[0091] The effect of the hydrogel composition of the present invention in alleviating inflammation around the alveolar bone and inducing tissue recovery was evaluated in guinea pigs with moderate to severe alveolar bone inflammation and abscess.

[0092] Specifically, in the subject guinea pig, abscess formation, mucosal redness, edema, and tissue damage were observed around the alveolar bone, and a void space with an empty interior due to the abscess was identified in the mandibular alveolar bone area. In addition, in the radiographic (X-ray) images taken before the application of hydrogel, irregular and increased penetrative osteolysis was observed in certain parts of the alveolar bone, along with a void formed by the abscess.

[0093] Regarding the alveolar bone inflammation site of the subject guinea pig, the area where the abscess had formed was first incised to drain the pus (drainage), and then the necrotic residual tissue was removed. Subsequently, 100 µl to 5 ml of hydrogel (410 µg / kg) prepared by the method of Example 1 was directly injected into the space formed after abscess removal and the alveolar bone area damaged by inflammation. The healing process, including the disappearance of inflammation, bone regeneration, abscess recurrence, and mucosal tissue condition, was observed for 13 days after hydrogel administration, and the results are presented in Table 3 and Figure 3.

[0094] Table 3 compares the clinical changes and bone regeneration status by category before and 13 days after administration of hydrogel in guinea pigs with induced alveolar bone inflammation and abscess, and Figure 3 is a photograph showing the hydrogel administration process and radiographic images and changes in the alveolar bone condition before and after administration in sequence.

[0095] Evaluation Criteria Before Hydrogel Administration 13 Days After Hydrogel Administration Abscess Large amount of abscess and pus present in the alveolar bone area Abscess disappeared, no pus or exudate observed Inflammation Severe redness and swelling of the gingiva and mucosa Reduction in redness and swelling, disappearance of inflammation Mucosa and Soft Tissue Condition Necrotic tissue and irregular mucosa observed Mucosa surface stabilization, epithelialization progressing Oral Hygiene Status Foreign matter and severe odor around the lesion Less area tidied up, odor disappearance Overall Healing Persistent inflammation and tissue damage progressing Distinct tissue recovery, gingival condition near normal

[0096] As shown in Table 3 and Figure 3, prior to hydrogel administration, a hollow space was observed in the alveolar bone area due to abscess formation and tissue damage, and radiographic images also revealed an irregular alveolar bone contour and hypodense areas suspected of bone resorption. Gross examination also clearly showed mucosal redness, edema, abscesses, and necrotic tissue. On the other hand, 13 days after hydrogel administration, radiographic images showed changes suggesting alveolar bone recovery, such as a reduction in the existing hollow space and a denser contour of the surrounding bone tissue. Gross examination also confirmed that the initial abscess and redness had mostly disappeared, the soft tissues around the tooth root had stabilized, and the mucosal surface had recovered uniformly. These results indicate that when the hydrogel composition of the present invention is applied locally to alveolar bone inflammation lesions, it reduces inflammatory exudate and abscesses and promotes alveolar bone regeneration in the defect area, and is effective in treating alveolar bone diseases accompanied by alveolar bone inflammation and alveolar bone defects.

Claims

1. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and A pharmaceutical composition for the prevention or treatment of inflammatory diseases, 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-βIGF-1, osteopontin, pleiotrophin, activin, endothelin-1, BDNF, GDNF, CNTF, cAMP, NT, A pharmaceutical composition that is 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. A pharmaceutical composition wherein the above-mentioned inflammatory disease is inflammatory bowel disease or periodontal disease.

11. In Claim 10, A pharmaceutical composition wherein the above-mentioned inflammatory bowel disease is one or more diseases selected from the group consisting of nonspecific chronic enteritis, indeterminate colitis, Crohn's disease, ulcerative colitis, microscopic colitis or subtypes thereof such as lymphocytic colitis, collagenous colitis, autoimmune enterocolitis, granulomatous enteritis, irritable bowel syndrome (IBS), and Behcet's colitis.

12. A pharmaceutical composition according to claim 10, wherein the periodontal disease is one or more diseases selected from the group comprising gingivitis, periodontitis, alveolar bone inflammation or alveolar bone defect, peri-implantitis, pulpitis, periapical abscess, stomatitis, mouth ulcers, and oral ulcers.

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

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

15. For use in the manufacture of medicines for the prevention or treatment of inflammatory 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.

16. For the treatment or prevention of inflammatory 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.

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