Composition for preventing or treating eye diseases comprising hydrogel

Abstract

The present invention relates to a composition or hydrogel for treating eye diseases. A composition or hydrogel according to one aspect forms a wet and viscoelastic protective film and secures retention when applied to a damaged area of an ocular surface (cornea, conjunctiva, etc.), thereby exhibiting the effects of inhibiting initial leakage and rapidly inducing re-epithelialization and tissue restoration. Accordingly, the present invention can be effectively used for preventing or treating eye conditions such as corneal epithelial defects, keratitis, ocular surface inflammation and trauma, wound healing after surgery, etc., and can be provided in ophthalmic formulations such as eye drop gels, eye ointments, patch-type hydrogels, etc.

Description

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

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

[0002] The eye is composed of intricate tissues such as the cornea, conjunctiva, iris, lens, and retina; in particular, the ocular surface is lubricated and protected by the tear film and extracellular matrix (ECM), which maintain epithelial homeostasis. Re-epithelialization of the corneal and conjunctival epithelium, structural stability of the ECM, and proper regulation of inflammatory responses are essential for maintaining vision and ensuring tissue transparency. However, if this balance is disrupted by trauma, surgery, dry eye, infection (keratitis / conjunctivitis), autoimmune or inflammatory diseases, or environmental stress, it can lead to epithelial defects, leakage, pain and hyperemia, chronic inflammation, and scar formation, resulting in a decline in visual function.

[0003] In such a state where the homeostasis of the ocular surface is disrupted, it is desirable to take an approach that rapidly seals the damaged area to prevent leakage, provides a moist and viscoelastic environment to support cell adhesion, migration, and differentiation to naturally accelerate re-epithelialization, and simultaneously induces the reconstruction of the ECM and the alleviation of inflammation.

[0004] However, existing eye drops and ointments tend to be limited to temporary lubrication and soothing effects; they also have limitations due to low retention on the ocular surface, a lack of growth factors and cell adhesion signals, and insufficient sustained tissue scaffolding, which delays re-epithelialization and structural restoration or leads to repeated re-injury. In particular, formulations that simultaneously satisfy the requirements for local scaffold-based microenvironment reconstruction, such as in traumatic corneal injury or post-surgical wounds, have not yet been sufficiently established.

[0005] Accordingly, the inventors have developed an ophthalmic hydrogel / composition by combining components having bio-support, cell adhesion, moisturizing, and viscoelastic properties. This composition is effective for the recovery of corneal and conjunctival damage and for the prevention and treatment of eye diseases by forming a moist and viscoelastic protective film at damaged areas of the ocular surface and ensuring retention to promote cell adhesion, migration, and survival.

[0006] One aspect provides a pharmaceutical composition for the prevention or treatment of eye disease 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.

[0007] Another aspect is to provide a pharmaceutical composition for the prevention or treatment of eye disease 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 an ophthalmic composition for the prevention or treatment of eye 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 an ophthalmic composition for the prevention or treatment of eye 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 eye 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 eye diseases.

[0012] Another aspect is to provide a hydrogel or hydrogel patch for the prevention or treatment of eye disease 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 eye disease 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.

[0014] Another aspect provides a composition for the prevention or treatment of eye disease 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 an ophthalmic composition for the prevention or treatment of eye 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 an ophthalmic composition for the prevention or treatment of eye 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 eye 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 eye 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 eye disease comprising the step of administering the above-mentioned 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 a formulation for preventing or treating eye diseases.

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

[0022]

[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 “ophthalmic diseases” in this specification is a term encompassing various pathological conditions resulting from structural or functional abnormalities of the eye, including cellular damage, inflammatory response, degeneration, or structural deformation in external and internal tissues constituting the eye, such as the ocular surface, cornea, conjunctiva, iris, lens, vitreous body, retina, optic nerve, orbit, and eyelid.

[0025] In one embodiment, the eye disease may be one or more selected from the group consisting of dry eye syndrome, conjunctivitis, keratitis, corneal ulcer, uveitis, glaucoma, cataract, retinal detachment, retinopathy, macular degeneration, optic neuritis, blepharitis, chalazion, hordeolum or stye, keratoconjunctivitis sicca, orbital cellulitis, optic neuropathy, exotropia, esotropia, ptosis, and ocular trauma, but is not limited thereto. no.

[0026] In one embodiment, the ocular trauma includes corneal abrasion / epithelial defect, corneal laceration (lamellar / full-thickness), corneal perforation, corneoscleral laceration, penetrating / perforating globe injury, open-globe rupture, iris prolapse, iris sphincter tear / iridodialysis, hyphema, angle recession, lens subluxation / dislocation, traumatic cataract, vitreous hemorrhage, traumatic retinal tear / detachment, It may be one or more selected from the group consisting of commotio retinae, choroidal rupture, eyelid laceration, canalicular laceration, conjunctival / corneal foreign body, chemical burns-alkali / acid, thermal / UV burns (including photokeratitis), orbital wall fracture (blow-out), retrobulbar hemorrhage, and traumatic optic neuropathy, but is not limited thereto.

[0027] In this specification, the term "pharmaceutical effective dose" may mean 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 pathological 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.

[0028] The terms “administering,” “applying,” “introducing,” 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 resulting in at least partial localization of the patch or composition according to one embodiment to a desired site.

[0029] 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.

[0030] 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.

[0031] In another embodiment, the hydrogel, hydrogel patch, or composition may be solid (including powder), semi-solid, or liquid. Additionally, for example, the 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.

[0032] 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.

[0033] In one embodiment, the hydrogel, hydrogel patch, or composition may include fibrin and / or fibrinogen.

[0034] The above fibrin acts as a major proteinaceous scaffold for wound healing and tissue regeneration processes in vivo, and can induce regeneration of the ocular surface and alleviate inflammation by promoting cell adhesion, cell migration, and growth factor delivery at damaged sites of the cornea, conjunctiva, or ocular surface.

[0035] In particular, the fibrin can contribute to the restoration of corneal transparency, the healing of epithelial defects, and the maintenance of visual function by promoting the adhesion and survival of corneal epithelial cells, keratocytes, and vascular endothelial cells, and by inducing angiogenesis and re-epithelialization of epithelial cells. Furthermore, the fibrin plays an important role in inflammatory modulation and tissue repair enhancement by adsorbing and releasing various growth factors (VEGF, PDGF, FGF, etc.) and cytokines.

[0036] The above fibrinogen is a precursor of fibrin and is converted into fibrin in vivo by the action of thrombin. Therefore, by including fibrinogen as a precursor drug and allowing fibrin to be gradually formed during the in vivo conversion process, wound healing and ocular surface regeneration can be induced to occur continuously and in a balanced manner. In other words, while the final pharmacological substance acting in vivo contains fibrin, fibrinogen can be used instead of fibrin as a precursor drug.

[0037] In addition, depending on the amount of substance that converts fibrin into fibrinogen or the degree of the fibrin formation reaction, the hydrogel, hydrogel patch composition, or ophthalmic hydrogel composition according to one embodiment may additionally include fibrinogen and / or thrombin.

[0038] By controlling the content of the fibrinogen and thrombin mentioned above, the density, mechanical strength, viscoelasticity, and biodegradation rate of the fibrin network can be controlled. Through this control of physical properties, intraocular retention and sustained release of the active ingredient can be improved, thereby enabling more efficient induction of tissue regeneration, such as in damaged cornea or conjunctiva.

[0039] Accordingly, the present specification may additionally provide a prodrug comprising fibrinogen; laminin; and / or hyaluronic acid. The 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.

[0040] 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.

[0041] 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.

[0042] In this specification, the term "laminin" refers to an extracellular matrix protein constituting the basal lamina, which may mean 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 above laminin is a major component of the basement membrane and is known to regulate cell adhesion, cell migration, differentiation, and cell survival by interacting with integrin receptors and heparan sulfate proteoglycans of the cell membrane.

[0043] The aforementioned laminin acts as a key protein constituting the basement membranes of ocular tissues, particularly the cornea, conjunctiva, and retina, and promotes the attachment and proliferation of epithelial cells, keratocytes, and retinal pigment epithelial cells. Through these actions, it induces re-epithelialization of the corneal epithelium and basement membrane reconstruction, and can promote the recovery from ocular surface damage and inflammatory eye diseases. In addition, the laminin can enhance the resistance to injury and regenerative capacity of corneal epithelial cells by activating the β1 integrin-ERK pathway and the PI3K / Akt signaling pathway to strengthen cell proliferation and survival signals, and can contribute to the restoration of damaged tissue and the maintenance of visual function in eye diseases such as corneal epithelial defect, keratitis, and dry eye syndrome.

[0044] 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.

[0045] In this specification, the term “hyaluronic acid” possesses excellent hygroscopicity and viscoelasticity and plays an important role in cell adhesion, migration, differentiation, and tissue remodeling as a major component of the extracellular matrix (ECM). In particular, within ocular tissues, said hyaluronic acid constitutes the ECM of the cornea, conjunctiva, and ocular surface and plays a key role in maintaining a stable intercellular microenvironment. The hyaluronic acid mentioned above improves the stability of the tear film through excellent moisturizing power and viscoelasticity, and improves the survival environment of corneal epithelial cells and conjunctival epithelial cells by alleviating tissue damage caused by dryness or mechanical friction. In addition, the hyaluronic acid induces extracellular matrix remodeling and re-epithelialization of epithelial cells, thereby promoting tissue regeneration and epithelial restoration at damaged sites of the cornea and conjunctiva. Through this, it maintains the lubrication and protective function of the ocular surface, inhibits the progression of ophthalmic diseases, and contributes to the maintenance of visual function.

[0046] The above hyaluronic acid is a type of glycosaminoglycan called hyaluronan, a polysaccharide with disaccharide bonds 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. 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.

[0047] In another embodiment, the 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, an epidermal growth factor, a hepatocyte growth factor, a transforming growth factor, or a combination thereof. More specifically, the growth factors may include placental growth factor, macrophage colony-stimulating factor, granulocyte macrophage colony-stimulating factor, neuropilin, fibroblast growth factor (FGF)-1, FGF-2 (bFGF), FGF-3, FGF-4, FGF-5, FGF-6, erythropoietin, BMP-2, BMP-4, BMP-7, BMP-9, TGF-beta, IGF-1, osteopontin, pleotropin, activin, endothelin-1, and combinations thereof. The above neuronal growth factor may include one or more selected from the group consisting of BDNF (Brain-derived neurotropic factor), GDNF (Glial cell derived neurotropic factor), CNTF (Ciliary neurotropic factor), bFGF (basic fibroblast growth factor), cAMP (cyclic adenocyne monophosphate), NT (Neurotropin), NT3 (Neurotropin-3), NT4 (Neurotropin-4), T3 (Triiodo-L-Thyronine), SHH (Sonic hedgehog), and PDGF (Platelet-derived growth factor). The above vascular endothelial growth factor may include vascular endothelial growth factor (VEGF)-A, VEGF-A, VEGF-B, VEGF-C, VEGF-D, or VEGF-E.

[0048] For example, the above cell growth factors include 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, It may be cAMP, NT, NT-3, NT-4, T3, SHH, PDGF, VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E or a combination thereof, but is not limited thereto.

[0049] The concentration of the cell growth factor included in the 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 the hydrogel patch or composition containing fibrin, laminin, and hyaluronic acid.

[0050] In other embodiments, the 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.

[0051] Additionally, the 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 patch or composition, or are not included at all. The 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.

[0052] In one embodiment, the 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 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.

[0053] 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.

[0054] 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.

[0055] 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.

[0056] 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.

[0057] The term “ophthalmic preparation” in this specification refers to a sterile pharmaceutical composition prepared for application to the eye or eyeball, and includes formulations intended for ophthalmic administration (topical administration to the surface of the eye), application into the conjunctival sac, and, if necessary, drug delivery to the anterior / posterior segment of the eye. The formulations may consist of solutions, suspensions, emulsions, gels (including hydrogels), ointments (ophthalmic ointments), inserts, etc. For example, the above ophthalmic preparation may include eye drops / solutions, suspensions, emulsions / microemulsions, ophthalmic gels / hydrogels (including temperature / pH / ion-induced in-situ gels), ophthalmic ointments, sprays / mists, foam formulations, hydrogel patches / ocular films, ocular inserts, punctal plugs (drug-eluting capable), contact lens-based delivery, and sustained-release depots / implants (subconjunctival, intravitreal, intracameral), and, if necessary, lipid bodies, niosomes, micelles, and nanoparticles. Nanocarriers such as micelles, nanoparticles, and nanogels can be applied to the above formulations to improve solubility, permeability, and sustained release, but are not limited thereto.

[0058] In addition, the ophthalmic formulation may include one or more pharmaceutically acceptable excipients (e.g., tonicity agents, buffers, viscosity modifiers / polymers, demulcants, surfactants, preservatives or chelating agents, stabilizers / solubilizers, lubricants, etc.) and may be combined in appropriate types and amounts according to the formulation purpose (protection / lubrication of the ocular surface, dissolution / dispersion / sustained release of the drug). In the case of an ophthalmic solution formulation, it is preferable that all components consist of water-soluble components that do not form precipitates or gel particles at the usual temperature and pH ranges and set concentrations during the preparation, sterilization, and storage of the formulation. Depending on the characteristics of the formulation, the ophthalmic formulation is sterilized by known methods such as filtration or heat sterilization, and may be provided as a single-dose formulation containing or without preservatives if necessary. If the above preservative is included, it may be formulated in an appropriate amount considering the safety and microbial control of ophthalmic devices (e.g., lenses) that come into contact with the formulation.

[0059] 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 ophthalmic preparations and personal hygiene products. When the above hydrogel, hydrogel patch, or composition is added to a quasi-drug composition for the purpose of preventing or improving eye diseases, the above 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).

[0060] 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.

[0061] The above method may also further include the step of incorporating a cell growth factor into the composition in the sol state.

[0062] 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.

[0063] 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.

[0064] The above hydrogel patch, fibrin and / or fibrinogen, laminin, hyaluronic acid, or cell growth factor are as described above.

[0065] 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.

[0066] When a composition or hydrogel according to one aspect is applied to a damaged area of ​​the ocular surface, such as the cornea or conjunctiva, it forms a moist and viscoelastic protective film and secures retention, thereby rapidly inducing re-epithelialization and tissue restoration, which is effective in treating eye diseases.

[0067] Figure 1 is a photograph showing the lesion condition before the application of hydrogel to clinical animals with traumatic corneal perforation and iris prolapse, and the structural recovery of the cornea and iris and the recovery of visual response 2 weeks after the application of hydrogel.

[0068] Figure 2 is a photograph and video capture screen showing the structural recovery of the cornea and iris and the recovery of visual response after application, comparing the changes before and after the application of the hydrogel of the present invention to a cat that had corneal perforation and iris prolapse and lost visual response due to traumatic corneal injury.

[0069] Figure 3 is a diagram showing the alleviation of lesions before and after the application of a hydrogel patch to clinical animals exhibiting chronic conjunctivitis and conjunctival prolapse.

[0070] Figure 4 is a photograph showing the structural recovery of the cornea and iris after application, comparing the changes before and two weeks after application of the hydrogel patch of the present invention in clinical animals that developed corneal perforation and iris prolapse due to ocular trauma.

[0071] The present invention will be explained in detail below through reference examples and embodiments. However, the following embodiments are merely illustrative of the present invention and should not be interpreted as limiting the present invention.

[0072] Example 1. Preparation of a hydrogel patch

[0073] 1.1. Hydrogel Formulation

[0074] 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.

[0075]

[0076] 1.2. Manufacture of Hydrogel Patch

[0077] 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 petridish. Subsequently, a hydrogel patch was prepared by curing at room temperature or 37°C through a sol-gel phase transition.

[0078]

[0079] Experimental Example 1. Confirmation of re-epithelialization and tissue recovery effects in traumatic corneal injury

[0080] The therapeutic effect of a hydrogel patch according to one embodiment of the present invention on eye diseases was evaluated in cats with confirmed traumatic corneal perforation and iris prolapse.

[0081] Specifically, perforation and iris prolapse were observed in the cornea of ​​the right eye of the subject cat, accompanied by inflammatory discharge and a pain response. 100 µl to 5 ml of hydrogel (410 µg / kg) prepared by the method of Example 1 was topically applied to the lesion site of the subject cat, and after application, a conjunctival flap was formed to cover and fix the defect site. No eye drops were administered during the period the flap was fixed. The progress was observed for 2 weeks after hydrogel application, and the results are shown in Table 1 and Figure 1.

[0082] Table 1 summarizes the progression of re-epithelialization, loss of leakage, and changes in inflammation and pain after applying hydrogel to the lesion, and Figure 1 is a photograph showing the condition of the lesion before applying hydrogel and the structural recovery of the cornea and iris and the recovery of visual response 2 weeks after applying hydrogel to clinical animals with traumatic corneal perforation and iris prolapse.

[0083] Evaluation Time | Regeneration / Tissue Status | Iris Location | Corneal / Ocular Inflammation | Visual Response Hydrogel Application | Anterior corneal perforation, iris prolapse, severe edema | External iris exposure | Hyperemia, edema, pain response | No visual response 2 weeks after hydrogel application | Regeneration and stabilization of corneal surface | Return to normal position | Disappearance of edema and hyperemia | Significant recovery of visual function

[0084] As shown in Table 1 and Figure 1, after the application of the hydrogel, the corneal perforation and iris prolapse lesions in cats showed structural stabilization within 2 weeks, corneal edema disappeared, and visual function responses reappeared. In addition, when the hydrogel patch of the present invention was used in combination with a conjunctival flap, it formed a moist and viscoelastic protective film, rapidly inducing re-epithelialization and tissue restoration. The fact that tissue healing proceeded smoothly despite the absence of eye drops during the treatment period suggests that the hydrogel acted as a protective barrier on the damaged ocular surface and promoted re-epithelialization. No significant side effects, such as irritation, excessive hyperemia, or abnormal pain, were observed throughout the entire course. These results indicate that the hydrogel patch prepared in Example 1 is effective for promoting re-epithelialization and tissue stabilization in ocular diseases such as traumatic corneal injury, and suggest that treatment continuity can be secured through retention and tissue protection functions even under conditions without the administration of eye drops.

[0085]

[0086] Experimental Example 2. Confirmation of therapeutic effect for corneal damage caused by glaucoma and bites

[0087] The therapeutic effect of the hydrogel according to one embodiment of the invention on eye diseases was evaluated in cats suffering from bilateral eye injuries caused by glaucoma and bite injury.

[0088] Specifically, the subject cat had underlying glaucoma in its right eye and an ocular perforation in its left eye caused by a bite injury during a fight with another cat. Before the application of the hydrogel patch, severe ocular damage accompanied by prolapse of the corneal and iris tissues was observed in the left eye, and there was almost no vision response to visual stimuli.

[0089] 100 µl to 5 ml of hydrogel (410 µg / kg) prepared by the method of Example 1 was applied topically to the lesioned eye of the above-mentioned cat, and subsequent observation of the visual response, stabilization of the eye structure, changes in activity, and changes in body weight were evaluated.

[0090] Table 2 summarizes the changes in visual response, ocular condition, and activity before and after the application of hydrogel to cats with traumatic corneal injury and loss of visual response.

[0091] Figure 2 is a photograph and video capture screen showing the structural recovery of the cornea and iris and the recovery of visual response after application, comparing the changes before and after the application of the hydrogel of the present invention to a cat that had corneal perforation and iris prolapse and lost visual response due to traumatic corneal injury.

[0092] The upper part of Fig. 2 illustrates the condition of the lesion prior to the application of the hydrogel, clearly showing (i) a corneal perforation, (ii) iris protrusion, and (iii) no vision response. Additionally, the upper part includes a photograph showing the procedure of applying the hydrogel patch to the damaged corneal area. The lower part of Fig. 2 illustrates the recovery state two months after the application of the hydrogel, confirming (i) structural stabilization of the corneal and conjunctival tissues, (ii) a clear recovery of the vision response, and (iii) the recovery of normal movement and behavior. The video capture screen includes footage of the cat's gaze tracking normally in response to visual stimuli (finger movements). The numbers displayed on the video capture screen represent sequential scene numbers according to the passage of time, and the arrows indicated within the images represent the finger movements (visual stimuli) presented to elicit a response from the cat.

[0093] At the time of evaluation Vision response Ocular / corneal condition Activity Before hydrogel patch application Almost no response to visual stimuli (no vision response) Left eye corneal perforation and iris prolapse, severe ocular injury Reduced activity due to glaucoma and pain, almost unable to move 2 months after hydrogel patch application Distinct response to visual stimuli observed (has vision response) Corneal and iris structures stabilized, perforation site healed Findings: Significant increase in activity, such as freely moving around the surrounding environment

[0094] As shown in Table 2 and Figure 2, as a result of applying the hydrogel patch of the present invention, visual function was restored and the ocular structure was stabilized in cats exhibiting severe traumatic ocular injury. These results suggest that the hydrogel composition of the present invention can be usefully employed to promote vision preservation and tissue recovery in ocular diseases accompanied by traumatic corneal and ocular injury.

[0095] Experimental Example 3. Confirmation of effect in improving conjunctivitis and conjunctival prolapse

[0096] The effect of the hydrogel prepared in Example 1 on improving eye diseases was evaluated in small dogs in which chronic conjunctivitis caused by persistent inflammation and prolapsed conjunctiva was observed because the conjunctival tissue failed to retract for several years after surgery.

[0097] Specifically, the subject small dog had a history of undergoing ophthalmic surgery related to the conjunctiva about 3 years ago, and after the surgery, the prolapsed conjunctiva failed to return to its normal position, resulting in a persistently protruding conjunctiva. Consequently, chronic ocular inflammation symptoms such as swollen conjunctiva, chronic hyperemia, increased mucous and serous discharge (ocular discharge), and epiphora were continuously observed.

[0098] 100 μl to 5 ml of hydrogel (410 μg / kg) prepared by the method of Example 1 was topically applied to the protruding conjunctival lesion surface of the subject small dog, and no eye drops or additional treatment were administered concurrently. The course was observed for 7 days after the application of the hydrogel, and the results are shown in Table 3 and Figure 3.

[0099] Table 3 summarizes the degree of lesion improvement by evaluation item after one week of application of a hydrogel patch to clinical animals with chronic conjunctivitis and conjunctival prolapse, and Figure 3 is a diagram showing the alleviation of lesions before and after application of a hydrogel patch to clinical animals with chronic conjunctivitis and conjunctival prolapse.

[0100] Evaluation Criteria Before Hydrogel Application After 1 Week of Hydrogel Application Reduction in Conjunctival Edema The conjunctiva was significantly protruding, and severe swelling was observed around the lesion. Swelling significantly decreased 1 week after application. Conjunctival Tissue Retraction The protruding (herniated) conjunctival tissue failed to return to its original position, and chronic hypertrophy was observed. The protruding conjunctiva mostly returned to its normal position. Relief from Inflammation Accompanied by hyperemia, inflammatory redness, and exudate. Hyperemia and inflammatory changes significantly decreased. Reduction in Discharge Mucous / purulent ocular discharge continuously observed. Mucous discharge significantly decreased compared to before. Pain Response Definite pain response upon palpation, avoidance of contact around the eyes observed. Pain response upon palpation disappeared. Functional Improvement Complaints of discomfort when opening and closing the eyes, persistent irritation / foreign body sensation, and difficulty opening the eyes. No foreign body sensation upon opening and closing the eyes, recovery of normal blinking function.

[0101] As shown in Table 3 and Figure 3, the hydrogel of the present invention showed effects of reducing conjunctival edema, alleviating inflammation, stabilizing conjunctival tissue, and reducing secretions within a short period (1 week) in small dogs with chronic conjunctivitis and conjunctival prolapse.

[0102] These results suggest that the hydrogel according to one embodiment of the present invention can be effectively used in the treatment of chronic conjunctival diseases by protecting conjunctival tissue, inhibiting inflammatory exudation, and creating an environment for tissue regeneration.

[0103]

[0104] Experimental Example 4. Confirmation of improvement effect on traumatic corneal perforation and iris tissue damage

[0105] The ocular tissue regeneration effect of the hydrogel prepared in Example 1 was evaluated in clinical animals, geckos, that had corneal perforation and iris tissue destruction due to traumatic injury.

[0106] Specifically, the subject lizard was observed to have severe corneal perforation, partial external exposure and necrosis of the iris tissue, severe inflammatory discharge, ocular depression, and pain response, and was in a severe condition clinically suspected of loss of visual function in the right eye.

[0107] 50 μl to 2 ml of a hydrogel patch (410 μg / kg) prepared by the method of Example 1 was directly applied to the corneal defect and exposed iris tissue area of ​​the above-mentioned lizard, and the degree of improvement in damage was evaluated at 2 weeks after the application of the hydrogel patch, and the results are shown in Table 4 and Figure 4.

[0108] Table 4 summarizes the degree of improvement in damage by category after applying a hydrogel patch to clinical animals that developed corneal perforation and iris prolapse due to ocular trauma, and Figure 4 is a photograph showing the structural recovery of the cornea and iris after application by comparing the changes before and 2 weeks after application of the hydrogel patch of the present invention to clinical animals that developed corneal perforation and iris prolapse due to ocular trauma.

[0109] Evaluation Criteria Before Hydrogel Patch Application After 2 Weeks of Hydrogel Patch Application Corneal Regeneration Presence of full-thickness corneal (perforation) damage, exposure of internal tissues (iris, etc.) at the defect site, loss of corneal surface morphology Closure of perforation site, restoration of corneal surface continuity Iris Regeneration Severe damage confirmed by external exposure and deformation of iris tissue, and redness of the tissue Disappearance of necrotic tissue and restoration of iris contour Reduction of Inflammation Presence of severe redness, swelling, and engorgement, distinct edema and inflammatory changes in surrounding soft tissues Significant reduction in redness and discharge Preservation of Visual Function Lack of visual response to visual stimuli, unobservable pupillary response Normal pupillary response (dazzle reflex) confirmed Tissue Stabilization Disintegration of ocular surface structure, ongoing additional tissue damage, high risk of infection Ocular surface structure stabilized, no additional damage

[0110] As shown in Table 4 and Figure 4, after the application of the hydrogel patch, the corneal perforation site was stabilized, regeneration of the corneal and iris structures was confirmed, and a state was observed in which the cornea recovered to a clear state and the axis of vision was maintained. These results indicate that the hydrogel patch of the present invention is effective for tissue regeneration and inflammation relief even in severe corneal damage in reptiles.

Claims

1. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and A pharmaceutical composition for the prevention or treatment of eye 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-β, IGF-1, osteopontin, pleiotrophin, activin, endothelin-1, BDNF, GDNF, CNTF, cAMP, A pharmaceutical composition that is NT, NT-3, NT-4, T3, SHH, PDGF, VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, or a combination thereof.

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

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

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

10. In Claim 1, A pharmaceutical composition wherein the above-mentioned eye disease is one or more selected from the group consisting of dry eye syndrome, conjunctivitis, keratitis, corneal ulcer, uveitis, glaucoma, cataract, retinal detachment, retinopathy, macular degeneration, optic neuritis, blepharitis, chalazion, hordeolum or stye, keratoconjunctivitis sicca, orbital cellulitis, optic neuropathy, exotropia, esotropia, ptosis, and ocular trauma.

11. In Claim 10, The above ocular trauma includes corneal perforation, iris prolapse, corneal abrasion / epithelial defect, corneal laceration (lamellar / full-thickness), corneoscleral laceration, penetrating / perforating globe injury, open-globe rupture, iris sphincter tear / iridodialysis, hyphema, angle recession, lens subluxation / dislocation, traumatic cataract, vitreous hemorrhage, and traumatic retinal tear / detachment. A pharmaceutical composition comprising one or more selected from the group consisting of commotio retinae, choroidal rupture, eyelid laceration, canalicular laceration, conjunctival / corneal foreign body, chemical burns-alkali / acid, thermal / UV burns (including photokeratitis), orbital wall fracture (blow-out), retrobulbar hemorrhage, and traumatic optic neuropathy.

12. Fibrin or, fibrin and fibrinogen; Laminin or, laminin-derived peptide or protein; and An ophthalmic composition for the prevention or treatment of eye diseases, comprising hyaluronic acid or a salt thereof as an active ingredient.

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

14. For use in the manufacture of medicines for the prevention or treatment of eye 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.

15. For the prevention or treatment of eye 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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