Composition for preventing or treating arthritis comprising hydrogel

Abstract

The present invention relates to a composition or hydrogel for preventing or treating arthritis. The composition or hydrogel according to one aspect effectively inhibits inflammatory responses occurring in cartilage, synovium, and articular cavities within joints, and can be applied to improve joint functions. In addition, the present invention can also be applied to inflammatory lesions in peripheral soft tissues supporting joints, such as tendons, ligaments, and bursae. Therefore, the composition or hydrogel of the present invention is effective for inflammatory diseases of the joint itself and various inflammatory lesions occurring in structures surrounding the joint.

Description

Composition for the prevention or treatment of arthritis comprising a hydrogel

[0001] This invention relates to a composition for the prevention or treatment of arthritis comprising a hydrogel.

[0002] Arthritis is a group of chronic diseases characterized by inflammatory reactions in the cartilage, synovium, joint capsule, and surrounding soft tissues (tendons, ligaments, bursae, etc.) that constitute the joint, causing pain, swelling, stiffness, limited range of motion, and tissue damage. As damage to intra-articular structures, inflammatory responses, and tissue degeneration progress in a complex manner, it leads to a decline in joint function and disability in daily life, and it occurs extensively in various parts of the body, including the knees, hips, ankles, hands, and spine.

[0003] The pathology of arthritis includes the loss of proteoglycans, a component of articular cartilage; degeneration and death of chondrocytes; synovitis; an increase in inflammatory mediators within the joint cavity; and inflammation and edema of surrounding soft tissues, which lead to the gradual deterioration of joint function. Meanwhile, inflammatory lesions frequently occur in surrounding tissues such as tendons, ligaments, and bursae, which are known to exacerbate joint pain and movement disorders.

[0004] Currently, non-steroidal anti-inflammatory drugs (NSAIDs), steroids, disease-modifying anti-inflammatory drugs (DMARDs), hyaluronic acid injections, and biological agents are widely used to treat inflammatory diseases of the joints and surrounding areas. However, systemically administered drugs can cause various systemic side effects with long-term use, such as gastrointestinal disorders, cardiovascular abnormalities, nephrotoxicity, and an increased risk of infection; furthermore, they have limitations in that it is difficult to maintain high concentrations of therapeutic agents within the joint cavity for extended periods. Intra-articular injections also often have a limited duration of effect or fail to sufficiently induce fundamental structural recovery of cartilage and joint tissues. Additionally, there are a significant number of cases where inflammatory lesions in the soft tissues surrounding the joints, such as tendons, ligaments, and bursae, recur or experience repeated symptoms despite conventional treatment.

[0005] As such, existing treatments have common limitations such as a lack of sustained efficacy, systemic side effects, poor local maintenance, and insufficient tissue regenerative capacity, and remain inadequate for fundamentally improving inflammatory lesions occurring in joints and surrounding tissues.

[0006] Based on this technical necessity, the inventors have developed a composition based on a composition comprising fibrinogen, laminin, and hyaluronic acid that can effectively suppress inflammation in joints and surrounding soft tissues and simultaneously induce tissue recovery and functional improvement. The composition of the present invention can complement the structural limitations of existing treatments by stably remaining within the joint cavity or around the joint, exerting a local anti-inflammatory effect, and contributing to the recovery of joint function.

[0007] One aspect provides a pharmaceutical composition for the prevention or treatment of arthritis 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 arthritis 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 arthritis 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 arthritis 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 arthritis 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 arthritis.

[0013] Another aspect is to provide a hydrogel or hydrogel patch for the treatment or prevention of arthritis 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 arthritis comprising fibrin and / or fibrinogen; laminin or a laminin-derived peptide or protein; and / or hyaluronic acid or a salt thereof as an active ingredient.

[0015] Another aspect provides a composition for the prevention or treatment of arthritis 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 arthritis 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 arthritis 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 arthritis 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 arthritis 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 arthritis 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 for the manufacture of pharmaceutical preparations for the prevention or treatment of arthritis.

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

[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 “arthritis” in this specification refers to a group of diseases characterized by acute or chronic inflammatory reactions occurring in joints and surrounding tissues as the primary lesion, and is characterized by structural changes in cartilage, synovium, ligaments, tendons, bursae, and surrounding soft tissues, as well as pain, edema, heat, range of motion reduction, and functional impairment. The above arthritis includes various inflammatory joint diseases caused by inflammation of the joint structure or immunological and metabolic abnormalities, such as osteoarthritis, rheumatoid arthritis (RA), osteoarthritis, psoriatic arthritis, gouty arthritis, ankylosing spondylitis, spondyloarthritis, reactive arthritis, enteropathic arthritis, lupus arthritis, and septic arthritis.

[0025] In one embodiment, the arthritis may include one or more diseases selected from the group consisting of osteoarthritis, rheumatoid arthritis (RA), osteoarthritis, psoriatic arthritis, gouty arthritis, ankylosing spondylitis, spondylitis, reactive arthritis, enteropathic arthritis, juvenile idiopathic arthritis (JIA), lupus arthritis, septic arthritis, bursitis, periarthritis of shoulder, tendinitis, tenosynovitis, paratendinitis, tenosynovial inflammation, myositis, or synovitis, but is not limited thereto. no.

[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.

[0033] 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.

[0034] 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.

[0035] 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.

[0036] 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.

[0037] 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.

[0038] 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. 6 Da, 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.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 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.

[0043] 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.

[0044] 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.

[0045] 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.

[0046] 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.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] 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 arthritis, the above hydrogel, hydrogel, hydrogel patch, or composition may be added as is or used together with other quasi-drug ingredients, or 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).

[0051] The above-mentioned quasi-drugs may include, but are not limited to, topical analgesic gels or lotions used for the purpose of temporarily relieving pain in joint areas, recovering from fatigue, providing hot or cold sensations, or alleviating local inflammatory reactions, such as linthia, medicated plaster, cooling / heating patch, topical analgesic gel or lotion, hydrocolloid patch for protecting or assisting the function of joint areas, external analgesic cream containing analgesic or anti-inflammatory ingredients, or topical spray for relieving muscle and joint fatigue.

[0052] 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.

[0053] The above method may also further include the step of incorporating a cell growth factor into the composition in the sol state.

[0054] 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.

[0055] 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.

[0056] The above hydrogel patch, fibrin and / or fibrinogen, laminin, hyaluronic acid, or cell growth factor are as described above.

[0057] 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.

[0058] The terms, methods, effects, etc. described for the above inventions apply equally between each invention.

[0059] According to one aspect, the above composition or hydrogel is effective in effectively suppressing inflammatory reactions occurring in the cartilage, synovium, and joint cavity within the joint and improving joint function. In addition, it is effective in treating inflammatory lesions occurring in surrounding soft tissues that support the joint, such as tendons, ligaments, and bursae. Therefore, the composition or hydrogel of the present invention is effective against inflammatory diseases of the joint itself and various inflammatory lesions occurring in structures surrounding the joint.

[0060] Figure 1 shows the results of evaluating the effect of a hydrogel according to one embodiment on improving arthritis symptoms in an animal model of rheumatoid arthritis.

[0061] Figure 2 is a graph showing the results of evaluating the cartilage degradation inhibitory effect of a hydrogel according to one embodiment in an animal model of degenerative arthritis.

[0062] Figure 3 shows the results of confirming the lesion site by radiographic imaging before and 5 days after administering a hydrogel according to one embodiment to clinical animals with induced hip arthritis, and confirming the severity through behavioral analysis (Lameness grade) of clinical animals by photographing their actual walking.

[0063] Figure 4 is a photograph comparing changes in lesions around the ankle before and 14 days after administration of a hydrogel according to one embodiment in clinical animals with induced ankle arthritis.

[0064] 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.

[0065]

[0066] Example 1. Preparation of a hydrogel patch

[0067] 1.1. Hydrogel Formulation

[0068] 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.

[0069] 1.2. Manufacture of Hydrogel Patch

[0070] 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.

[0071] Experimental Example 1. Confirmation of the arthritis treatment effect of a hydrogel patch

[0072] 1.1 Confirmation of Treatment Efficacy for Rheumatoid Arthritis

[0073] 1.1.1. Preparation of an Animal Model for Rheumatoid Arthritis

[0074] To confirm the effect of the hydrogel of the present invention on improving rheumatoid arthritis (RA), an animal model of rheumatoid arthritis was prepared and disease activity was evaluated.

[0075] Specifically, male DBA / 1J mice aged 7 weeks were used to create a rheumatoid arthritis model, and experiments were conducted under isoprene anesthesia. A solution of 100-200 µl containing 2 mg / ml of collagen type II mixed with an equal amount of complete Freund's adjuvant was injected once subcutaneously into the skin of the mouse's tail, and collagen was injected in the same manner 2 weeks later to create a collagen-induced arthritis (CIA) animal model.

[0076]

[0077] 1.1.2. Verification of Arthritis Recovery Effect through Confirmation of Arthritis Clinical Indices

[0078] For mice with rheumatoid arthritis induced according to the method of Experimental Example 1.1.1 above, the degree of swelling in the ankle and knee joints of both hind feet after collagen induction was evaluated at one-week intervals. Specifically, the hydrogel of the present invention or physiological saline (control group) was injected into the joint cavity or peri-joint area of ​​the mouse in the rheumatoid arthritis animal model constructed in Experimental Example 1.1.1, and the clinical index of the mouse was analyzed up to 7 weeks after injection.

[0079] The clinical severity of rheumatoid arthritis was evaluated based on the hind feet of mice, and a Clinical Arthritis Score (CIA Score) was calculated by synthesizing clinical changes observed in each individual, such as the extent of edema, redness, and joint stiffness. The evaluation criteria for the above clinical score are as follows.

[0080] - 0 points: Normal, no redness or swelling

[0081] - 1 point: Mild swelling / redness, swelling of 1-2 toes or localized edema on the top of the foot

[0082] - 2 points: Moderate swelling, swollen 3 or more toes or slight swelling of the entire foot

[0083] - 3 points: Severe edema, joint stiffness, generalized swelling of the foot / ankle, reduced movement

[0084] - 4 points: Severe swelling and inability to grip / hold the toes, joint deformity

[0085] Figure 1 shows the results of evaluating the degree of progression of rheumatoid arthritis and the effect of the hydrogel on improving arthritis symptoms based on the above clinical index.

[0086] Figure 1 shows the results of evaluating the effect of a hydrogel according to one embodiment on improving arthritis symptoms in an animal model of rheumatoid arthritis.

[0087] The left panel of FIG. 1 illustrates an experimental schedule in which rheumatoid arthritis (CIA) is induced through primary and secondary immunity of collagen (type II collagen), and the hydrogel of the present invention is injected into the joint cavity or peri-joint area at the time when CIA is established (week 3), and clinical indices are evaluated for a certain period.

[0088] The graph on the right in Figure 1 compares the changes in the clinical arthritis index of the Sham group, CIA group, and hydrogel administration group (CIA-Hydrogel) over time (Weeks 0-10).

[0089] As shown in Figure 1, in the CIA group, joint swelling and redness gradually worsened after immune induction, with the clinical index reaching its peak at weeks 5–6, whereas in the hydrogel administration group, a trend of a significant decrease in the arthritis clinical index was observed after the same time point.

[0090] These results indicate that the hydrogel of the present invention effectively alleviates inflammatory lesions such as joint swelling, redness, and joint stiffness in animal models of rheumatoid arthritis, and exhibits therapeutic efficacy in improving arthritis-related symptoms.

[0091]

[0092] 1.2. Confirmation of therapeutic effect for degenerative arthritis

[0093] 1.2.1 Preparation of an Animal Model for Degenerative Arthritis

[0094] Forty 7-week-old white rats [Sprague Dawley (SD) rats] were divided into 5 groups of 8 rats each, placed in cages, and stabilized for one week.

[0095] After thoroughly shaving the area around the right knee joint of white rats, osteoarthritis was induced by injecting 50 uL (40 mg / mL) of MIA (Sig-ma-Aldrich Co., St. Louis, MO, USA), dissolved in 0.85% physiological saline, into the joint cavity, while the same amount of physiological saline was injected into the normal (Sham) group instead of MIA. After 24 hours, hydrogel, ibuprofen, or physiological saline (control group) was administered.

[0096]

[0097] 1.2.2 Inhibition of Cartilage CAG Degradation: Confirmation of Increase in Collagen Concentration

[0098] Proteopolysaccharides, which are a major component of the tissues that make up the joint, are a representative extracellular matrix material in which one to several GAGs are covalently bonded to a core protein. As osteoarthritis progresses, GAGs are degraded and their concentration decreases. Through experiments, we confirmed whether a hydrogel inhibits the degradation of GAGs in the cartilage of an animal model of osteoarthritis induced by MIA.

[0099] Specifically, a hydrogel, ibuprofen, or physiological saline (control group) was administered to the animal model of degenerative arthritis prepared in Experimental Example 1.2.1. After 8 weeks, white rats were anesthetized with ether, sacrificed, and the right knee cartilage was excised. PBS was added to the excised cartilage and homogenized using a homogenizer, followed by centrifugation at 10,000 rpm for 10 minutes to obtain the supernatant. The concentration of glycosaminoglycan (GAG) in this supernatant was determined by measuring at 450 nm using a microplate spectrophotometer with an ELISA kit. The concentration of collagen was obtained by adding 100 µL of the sample obtained above and 100 µL of 6 N HCl to each well, hydrolyzing at 121 ℃ for 3 hours, centrifuging at 10,000 rpm for 10 minutes, and measuring at 560 nm using BioVision's hydroxyproline colorimetric assay kit, and the results are shown in Figure 2.

[0100] Figure 2 is a graph showing the results of evaluating the cartilage degradation inhibitory effect of a hydrogel according to one embodiment in an animal model of degenerative arthritis. All data are expressed as mean ± standard deviation (mean ± SD), Sham refers to the normal group, Control refers to the group administered physiological saline to the group of degenerative arthritis induced by MIA, Ibuprofen refers to the group administered ibuprofen to the group of degenerative arthritis induced by MIA, and Hydrogel refers to the group of degenerative arthritis induced by MIA administered hydrogel.

[0101] As shown in Figure 2, the total protein was measured at 27.32±3.36 ng / mg in the normal group (Sham) and 20.43±2.28 ng / mg in the control group, indicating that the GAG ​​concentration in the cartilage was reduced in the group that induced osteoarthritis with MIA.

[0102] In addition, the ibuprofen and hydrogel administration groups showed significantly increased GAG concentrations compared to the control group, with 28.74 ±3.23 ng / mg total protein, 29.73 ±2.23 ng / mg total protein, 31.39 ±2.42 ng / mg total protein, 30.83 ±1.99 ng / mg total protein, and 32.48 ±2.28 ng / mg total protein, respectively.

[0103] These results indicate that the hydrogel inhibits GAG degradation in an animal model of osteoarthritis induced by MIA, and that it has an effect of alleviating the progression of osteoarthritis.

[0104] Meanwhile, changes in collagen concentration in cartilage tissue after MIA induction were measured, and the results are listed in Table 1.

[0105] Collagen (μg / mg) Sham (Normal Group) DW (MIA Control Group) Ibuprofen Hydrogel Mean ± SD 5.97 ± 2.0 14.17 ± 0.9 5 5.57 ± 2.0 15.35 ± 3.04

[0106] As shown in Table 1, collagen concentrations were 5.97±2.01 μg / mg total protein in the normal group (Sham) and 4.17±0.95 μg / mg total protein in the control group; it was confirmed that arthritis was induced because the collagen concentration in the control group was lower than that in the normal group (Sham). Meanwhile, in the groups administered ibuprofen and hydrogel, the values ​​were measured as 5.57±2.01 μg / mg total protein, 5.35±3.04 μg / mg total protein, 5.57±2.70 μg / mg total protein, 5.47±2.06 μg / mg total protein, and 5.97±1.70 μg / mg total protein, respectively.

[0107] These results mean that the hydrogel patch according to one embodiment of the present invention is effective in treating arthritis.

[0108]

[0109] 1.3. Confirmation of Arthritis Improvement Effect in Animal Clinical Cases

[0110] 1.3.1 Confirmation of Pain Relief and Gait Restoration Effects in Hip Arthritis

[0111] The effects of a hydrogel according to one embodiment of the present invention on joint pain relief and walking function improvement were evaluated on canine clinical animals suffering from hip osteoarthritis.

[0112] Specifically, the subject medium-sized dog had significantly narrowed joint spaces in both hips, making normal walking difficult due to severe pain, and exhibited persistent limping even when moving. Radiographic examination of the subject medium-sized dog prior to hydrogel administration revealed that the joint space between both femoral heads and acetabulums was almost completely lost, which indicates the progression of severe arthritis.

[0113] 200 µl to 5 ml of hydrogel (410 µg / kg) prepared by the method of Example 1 was administered to the hip joints of the subject medium-sized dog, and walking function, signs of pain, range of motion, weight-bearing capacity, and radiological changes were evaluated while observing for 5 days after administration, and the results are shown in Table 2 and Figure 2.

[0114] Table 2 summarizes the degree of improvement in hip arthritis by evaluation item after administering hydrogel to clinical animals with induced hip arthritis, and Figure 3 shows the results of confirming the lesion site by radiography before and 5 days after administering the hydrogel according to one embodiment to clinical animals with induced hip arthritis, as well as confirming the severity through behavioral analysis (Lameness grade) of the clinical animals by filming their actual gait. As a result of the behavioral analysis before and after hydrogel treatment, it took 5 days to measure the time for the Lameness grade to change from 5 to 1, and the result is as shown in Figure 2. Through these results, it was confirmed that the hydrogel has a therapeutic effect on arthritis.

[0115] Evaluation Criteria Observation Results 5 Days After Hydrogel Administration Pain Reduction: Significant reduction in pain response 5 days after administration Recovery of Walking Function: Disappearance of limping, ability to walk normally Increase in Joint Range of Motion: Confirmed increase in hip flexion and extension range Improvement in Weight-Bearing Capacity: Recovery of hind limb weight-bearing, disappearance of abnormal posture Imaging Improvement: Joint space partially recovered compared to pre-administration levels

[0116] As shown in Table 2 and Figure 2, after administration of the hydrogel, small dogs suffering from hip osteoarthritis showed a significant reduction in limping during walking. Although they had difficulty moving due to hind leg pain before administration, they were able to walk normally during movement and daily activities 5 days after administration. In addition, radiographic images confirmed that the hip joint space, which had been almost completely lost before administration, improved compared to before administration. This is attributed to the reduction of inflammation and tissue relaxation effects in the joint area. These results indicate that the hydrogel of the present invention demonstrates therapeutic effects of pain relief, recovery of joint function, and improvement of walking in animals with hip osteoarthritis.

[0117]

[0118] 1.3.2 Confirmation of Effects on Reducing Local Edema and Pain in Ankle Arthritis

[0119] The effect of a hydrogel according to one embodiment of the present invention on reducing local swelling and pain was evaluated in clinical animals suffering from ankle joint arthritis.

[0120] Specifically, the subject small dogs exhibited clinical symptoms such as local swelling, heat, pain on palpation, and lameness due to ankle joint arthritis, and no significant improvement was observed despite long-term physical therapy and general treatment. Prior to the administration of hydrogel, significant soft tissue swelling around the ankle, pain response to pressure, and local discoloration were observed.

[0121] 200 µl to 5 ml of hydrogel (410 µg / kg) prepared by the method of Example 1 was injected into the intra-articular of the subject small dog, and changes in clinical symptoms were observed for 14 days after administration of the hydrogel according to the evaluation items of Table 3, and the results are shown in Table 3 and Figure 3.

[0122] Table 3 summarizes the degree of improvement in ankle arthritis by evaluation item after administration of hydrogel to clinical animals with induced ankle arthritis, and Figure 4 is a photograph comparing changes in lesions around the ankle before and 14 days after administration of hydrogel to clinical animals with induced ankle arthritis.

[0123] Evaluation Criteria Observation Results After Hydrogel Administration Reduction in Edema: Significant edema around the ankle joint largely disappeared. Alleviation of Inflammatory Response: Fever and inflammatory erythema returned to normal levels. Reduction in Pain: Pain response upon palpation was significantly reduced. Normalization of Skin Tone: Skin tone in the affected area recovered to a level similar to normal skin. Recovery of Joint Function: Reduced limping during walking and improved range of motion. Improvement of Daily Activities: Signs of pain disappeared during activity, enabling normal walking.

[0124] As shown in Table 3 and Figure 4, about 14 days after administration of the hydrogel, ankle joint swelling was significantly reduced, and skin tone and local temperature returned to the normal range. Pain responses during walking were also significantly alleviated, and it was confirmed that weight-bearing and walking functions were restored to near-normal levels. With these results, it was confirmed that the hydrogel of the present invention is effective in suppressing local inflammation, alleviating swelling, reducing pain, and restoring joint function in clinical animals with ankle arthritis.

Claims

1. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and A pharmaceutical composition for the prevention or treatment of arthritis, 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. In Claim 1, A pharmaceutical composition wherein the above arthritis is one or more diseases selected from the group consisting of osteoarthritis, rheumatoid arthritis (RA), osteoarthritis, psoriatic arthritis, gouty arthritis, ankylosing spondylitis, spondylitis, reactive arthritis, enteropathic arthritis, juvenile idiopathic arthritis (JIA), lupus arthritis, septic arthritis, bursitis, periarthritis of shoulder, tendinitis, tenosynovitis, paratendinitis, tenosynovial inflammation, myositis, and synovitis.

11. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and A quasi-drug composition for the prevention or treatment of arthritis, 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 arthritis 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 arthritis, 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.

14. For the treatment or prevention of arthritis, 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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