Novel cerium oxide nanocomposite with enhanced biostability and use thereof

Abstract

The present invention relates to a cerium oxide nanocomposite, a method for preparing same, and a composition for the prevention or treatment of inflammatory or autoimmune diseases, comprising same as an active ingredient. The present invention can be used as an excellent therapeutic composition in which both biostability and in vivo efficiency of scavenging reactive oxygen species are maximized by modifying the surface of cerium oxide nanoparticles with an optimal amount of a pyrrolidone derivative polymer. Also, the method for preparing the nanocomposite, of the present invention, can form uniform particles having an optimal diameter while completely removing toxicity caused by reaction residues and nitrate by only a simple process of washing with a sodium chloride solution having a specific concentration. The present invention can be usefully applied to an excellent therapeutic composition which significantly inhibits inflammatory responses in various tissues, and particularly, minimizes damage to nerve cells, caused by excessive inflammatory responses in the periphery of hematoma due to intracerebral hemorrhage, thereby restoring neurological functions and greatly improving the survival rate of patients.

Description

Novel cerium oxide nanocomposite with enhanced biostability and its use

[0001] The present invention relates to a novel cerium oxide nanocomposite having significantly improved biostability while simplifying the production process, and a composition for preventing or treating various inflammatory diseases comprising the same as an active ingredient.

[0002]

[0003] Recently, active research is being conducted on nanoparticle-based therapeutics as pharmacological agents for various intractable diseases. Nanoparticles are being studied in various medical fields, including imaging diagnostics, drug delivery systems, and therapeutics, based on their unique optical, electromagnetic, and chemical properties stemming from their minute nanoscale diameters. In particular, nanoparticles can be utilized as pharmacological agents themselves, based on their magnetic properties, such as heat generation and antioxidant activity. Furthermore, active efforts are being made to develop nanoparticles that exhibit optimal properties for diverse medical applications by tailoring their composition, shape, size, and properties to suit specific purposes.

[0004] Reactive oxygen species (ROS), which are naturally produced by oxygen metabolism, are essential for life due to their functions such as cell signaling and immune response, but excessively produced ROS cause various oxidative stresses by inducing oxidative damage to biomolecules and cells. Cerium oxide nanoparticles, also called ceria nanoparticles, are thermally stable at high temperatures and can react with oxygen concentrations around them due to their lattice structure. 4+ / Ce 3+ It has various applications such as electrolytes for solid batteries, materials for UV filters, oxygen sensors, and optical devices due to its redox action.

[0005] In the medical field, cerium oxide nanoparticles are also attracting attention as therapeutic compositions for a wide range of inflammatory diseases. Cerium oxide nanoparticles reversibly bind to oxygen and form Ce on the particle surface. 3+ and Ce 4+ It can effectively remove free radicals by changing the oxidation state of the liver, so it can not only effectively control secondary brain damage caused by brain tissue inflammation due to various causes, but also function as a pharmacological ingredient for various diseases (e.g., dermatitis, inflammatory bowel disease, Crohn's disease, acute respiratory distress syndrome, inflammatory liver disease, etc.) caused by excessive inflammatory response in skin, intestine, lung, liver tissue, etc. However, due to the characteristics of nanoparticles with a fine diameter, it is very difficult to suppress aggregation during mass production and maintain uniform particle size, excellent dispersibility, and low surface charge (Zeta-potential). Therefore, there is a need for the development of novel cerium oxide nanoparticles with excellent antioxidant effect and optimized particle characteristics even in industrial-scale production to secure a therapeutically effective amount.

[0006]

[0007] Numerous papers and patents are referenced and cited throughout this specification. The disclosures of these cited papers and patents are incorporated herein by reference in their entirety to provide a clearer understanding of the state of the art and the scope of the present invention.

[0008]

[0009] The present inventors have diligently researched and strived to develop a nanoparticle-based therapeutic composition that can effectively treat various inflammatory diseases caused by tissue damage due to oxidative stress. As a result, the inventors discovered that when a pyrrolidone polymer of the following chemical formula 1 is bonded to the surface of cerium oxide nanoparticles at a certain content to form an outer layer, the dispersibility of the nanoparticles is significantly improved and the surface charge converges to 0 mV, thereby maximizing both biostability and the efficiency of removing reactive oxygen species in the body, and thus the present invention was completed.

[0010] Accordingly, the purpose of the present invention is to provide a novel cerium oxide nanocomposite and a method for producing the same.

[0011] Another object of the present invention is to provide a composition for preventing or treating inflammatory or autoimmune diseases, comprising the cerium oxide nanocomposite of the present invention as an active ingredient.

[0012]

[0013] Other objects and advantages of the present invention will become more apparent from the detailed description, claims and drawings below.

[0014]

[0015] According to one aspect of the present invention, the present invention provides a cerium oxide nanocomposite comprising:

[0016] (a) a core layer of cerium oxide nanoparticles; and

[0017] (b) an outer layer comprising a polymer represented by the following chemical formula 1:

[0018] Chemical Formula 1

[0019]

[0020] In the above chemical formula, R1 and R2 are each independently hydrogen or oxygen, represents a single bond or a double bond, l is 1 or 2, and m is an integer from 100 to 1000.

[0021] The present inventors have conducted extensive research efforts to develop a nanoparticle-based therapeutic composition that can effectively treat various inflammatory diseases caused by tissue damage due to oxidative stress. As a result, we discovered that when a pyrrolidone polymer of Chemical Formula 1 is bonded to the surface of cerium oxide nanoparticles at a certain content to form an outer layer, the dispersibility of the nanoparticles is significantly improved and the surface charge converges to 0 mV, thereby maximizing both biostability and the efficiency of removing reactive oxygen species in the body, and can be used as an excellent therapeutic composition.

[0022] In this specification, the term “core layer” means the innermost layer in a multilayer composite having only one surface in contact with another layer.

[0023] As used herein, the term “multilayer composite” refers to a composite composed of multiple layers composed of different components, and includes, without limitation, laminated multilayer structures, core-shell multilayer structures, and combinations thereof. Specifically, the multilayer composite of the present invention is a core-shell multilayer structure in which a nanoparticle is present at the center and a polymer of chemical formula 1 surrounds the nanoparticle at the outer surface.

[0024] The term “outer layer” as used herein refers to a layer surrounding the core layer of a core-shell structure and located further from the center than the core layer. The outer layer need not necessarily be a layer in direct contact with the core layer, and there may be an additional layer (e.g., an inner layer) that is closer to the core layer than the outer layer. The outer layer also need not necessarily be the outermost layer, and there may additionally be an outermost layer located further from the core than the outer layer.

[0025] According to the present invention, the boundary between the core layer and the outer layer may or may not be clearly distinct. If the boundary between the core layer and the outer layer is not clear, each component may be mixed at the boundary area or throughout the entire area between the two layers.

[0026] As used herein, the term “polymer” refers to a synthetic or natural polymer compound in which monomers of the same or different types are continuously bonded. Accordingly, the polymer includes a homopolymer (a polymer formed by polymerizing one type of monomer) and a copolymer prepared by polymerizing at least two different monomers, and the copolymer includes both a copolymer (a polymer prepared from two different monomers) and a polymer prepared from more than two different monomers. Specifically, the polymer of Chemical Formula 1 used in the present invention is a homopolymer.

[0027] As used herein, the term “alkyl” means a straight-chain or branched saturated hydrocarbon group, and includes, for example, methyl, ethyl, propyl, isopropyl, etc. C1-C3 alkyl means an alkyl group having an alkyl unit having 1 to 3 carbon atoms, and when C1-C3 alkyl is substituted, the carbon number of the substituent is not included.

[0028] In this specification, the term “biocompatibility” refers to a property of not causing short-term or long-term side effects when administered into a living body and comes into contact with cells, tissues or body fluids of an organ. Specifically, it includes tissue compatibility and blood compatibility, which do not cause necrosis of tissue or coagulation of blood when in contact with living tissue or blood, as well as biodegradability, which disappears after a certain period of time after administration into a living body, and “the property of being excreted from the body without accumulation after administration into a living body (excretability).” Therefore, the term “biocompatible dispersion stabilizer” refers to a component that has the above-described biocompatibility and improves the dispersibility of particles.

[0029] In this specification, the term “biodegradability” means a property of being naturally decomposed when exposed to a physiological solution of pH 6-8, and specifically means a property of being able to be decomposed over time by body fluids, decomposing enzymes, or microorganisms in a living body.

[0030] According to a specific embodiment of the present invention, the cerium oxide nanoparticles used in the present invention may be selected from the group consisting of cerium(III) oxide (Ce2O3) nanoparticles, cerium(IV) oxide (CeO2) nanoparticles, and mixtures thereof.

[0031]

[0032] According to a specific embodiment of the present invention, in the chemical formula 1, R1 is hydrogen, R2 is oxygen, and l is 1. According to the octet rule, when R1 is hydrogen, is a single bond and R2 is oxygen It is obvious that it is a double bond. The compound with chemical formula 1, where R1 is hydrogen, R2 is oxygen, and l is 1, is polyvinylpyrrolidone (PVP).

[0033]

[0034] According to a specific embodiment of the present invention, the polymer represented by the chemical formula 1 and the cerium oxide nanoparticles have a content ratio of 7:1 to 11:1.

[0035] According to the present invention, the inventors of the present invention have observed that when the polymer of chemical formula 1 and the cerium oxide nanoparticles have a content ratio of 7:1 to 11:1 (7 - 11 mg PVP / mg Ce) in the nanocomposite of the present invention, the nanocomposite exhibits the best particle characteristics by showing optimal biostability, dispersibility, and maximized inflammation control effect, thereby confirming that the biocompatibility and therapeutic effect are significantly improved compared to the conventional cerium oxide nanocomposite that requires additional modification of the outermost layer with a dispersion stabilizer such as PGA (polyglutamic acid), while the efficiency and economy of mass production through simplification of the structure can be drastically improved.

[0036] More specifically, the polymer represented by the above chemical formula 1 and the above cerium oxide nanoparticles have a content ratio of 7.5:1 to 10.9:1, even more specifically, a content ratio of 8:1 to 9.5:1, even more specifically, a content ratio of 8.5:1 to 9:1, and most specifically, a content ratio of about 8.7:1.

[0037] According to a specific embodiment of the present invention, the nanocomposite of the present invention additionally includes a multifunctional ligand represented by the following chemical formula 2:

[0038] Chemical Formula 2

[0039]

[0040] In the above chemical formula, n is an integer from 3 to 7.

[0041] The term “multi-functional ligand” as used herein refers to a molecule having two or more active functional groups and acting as a linker between two or more molecules by binding to them. The multi-functional ligand of Chemical Formula 2 used in the present invention has a carboxyl group capable of binding to cerium oxide nanoparticles and an amine group capable of binding to a compound of Chemical Formula 1 (e.g., PVP) of the outer layer, thereby enabling the nanocomposite of the present invention to be formed more efficiently and stably. Therefore, the multi-functional ligand of Chemical Formula 2 may be present within the outer layer or between the core layer and the outer layer, and in this case, the carboxyl group of the multi-functional ligand may be aligned in a direction in which it faces the core, and the amine group faces the compound of Chemical Formula 1 (Fig. 1a).

[0042] Specifically, in the above chemical formula 2, n is 5. The compound of chemical formula 2 where n is 5 is 6-aminohexanoic acid (6-AHA).

[0043]

[0044] According to a specific embodiment of the present invention, the nanocomposite of the present invention has an average particle diameter of 5 nm to 80 nm. More specifically, it has an average particle diameter of 10 nm to 50 nm, even more specifically, it has an average particle diameter of 15 nm to 30 nm, and most specifically, it has an average particle diameter of about 20 nm.

[0045] According to a specific embodiment of the present invention, the nanocomposite of the present invention has a surface charge (Zeta potential, mV) of -1.0 to 1.0 mV, and more specifically, a surface charge (Zeta potential, mV) of -0.5 to 0.5 mV. According to the present invention, the nanocomposite of the present invention exhibits a remarkably low surface charge that converges to 0 mV, thereby minimizing interaction with the aqueous biological environment when injected into the human body, thereby exhibiting excellent biostability.

[0046]

[0047] According to another aspect of the present invention, the present invention provides a composition for preventing or treating inflammatory or autoimmune diseases, comprising the nanocomposite of the present invention as an active ingredient.

[0048] According to another aspect of the present invention, the present invention provides a method for preventing or treating an inflammatory or autoimmune disease, comprising administering to a subject the nanocomposite of the present invention described above.

[0049] Since the cerium oxide nanocomposite used in the present invention has already been described above, its description is omitted to avoid excessive duplication.

[0050] The term “prevention” as used herein means inhibiting the occurrence of a disease or condition in a subject who has not been diagnosed as having the disease or condition but is susceptible to such disease or condition.

[0051] As used herein, the term “treatment” means (a) inhibiting the development of a disease, condition, or symptom; (b) alleviating the disease, condition, or symptom; or (c) eliminating the disease, condition, or symptom. The composition of the present invention acts to inhibit the development of symptoms caused by excessive or unwanted immune response or inflammation by reducing reactive oxygen species, eliminating them, or alleviating them. Specifically, as shown in the examples below, the composition of the present invention significantly improves the survival rate of individuals suffering from severe cerebral infarction and significantly reduces the volume of infarcted lesions. Therefore, the composition of the present invention may be a composition for treating inflammatory or autoimmune diseases, specifically cerebral infarction, on its own, or may be administered together with other pharmacological ingredients and applied as an adjuvant treatment for the above diseases. Accordingly, the terms “treatment” or “therapeutic agent” as used herein include the meaning of “adjuvant treatment” or “adjuvant treatment agent.”

[0052] The term “administration” as used herein means directly administering a therapeutically effective amount of the composition of the present invention to a subject so that the same amount is formed in the body of the subject, and has the same meaning as “transplantation” or “infusion.”

[0053] In the present invention, the term “therapeutically effective amount” means the content of the composition contained in an amount sufficient to provide a therapeutic or preventive effect to an individual to whom the composition of the present invention is to be administered, and includes “prophylactically effective amount”.

[0054] The term “subject” as used herein includes, without limitation, a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, monkey, chimpanzee, baboon, or rhesus macaque. Specifically, the subject of the present invention is a human.

[0055] According to a specific embodiment of the present invention, the inflammatory or autoimmune diseases prevented or treated by the composition of the present invention include intracerebral hemorrhage (ICH), cerebral infarction, dermatitis, traumatic brain injury, inflammatory bowel disease (IBD), and subarachnoid hemorrhage.SAH), cerebral edema, hypoxic ischemic encephalopathy, stroke, traumatic spinal cord injury, gunshot injury, acute respiratory distress syndrome, cytokine storm syndrome, sepsis, systemic inflammatory response syndrome, peritonitis, multiple sclerosis, rheumatoid arthritis, reactive arthritis, type 1 diabetes, systemic lupus erythematosus, idiopathic pulmonary fibrosis, polymyositis, dermatomyositis, localized scleroderma, systemic scleroderma, Sjogren's syndrome, Raynaud's phenomenon, Bechet's disease, Kawasaki's disease, primary biliary cirrhosis selected from the group consisting of biliary sclerosis, primary sclerosing cholangitis, psoriasis, myasthenia gravis, autoimmune vasculitis, primary angiitis of the central nervous system, and inflammatory liver disease;

[0056]

[0057] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is intracerebral hemorrhage (ICH).

[0058] In this specification, the term “intracranial hemorrhage (ICH)”, also called “intracranial hemorrhage,” refers to a disease in which brain tissue is contaminated and damaged by blood leaking due to damage to a cerebral blood vessel. Physical brain tissue damage occurs primarily due to the volume of the hematoma created by blood leakage from the cerebral blood vessel, and an inflammatory response is subsequently induced by the resulting cytotoxic substances such as reactive oxygen species, heme, and thrombin, which causes edema around the hematoma. Therefore, in order to suppress the symptoms of intracerebral hemorrhage, it is important to quickly suppress the edema and inflammatory response around the hematoma to prevent early neuronal damage caused by them.

[0059]

[0060] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is cerebral infarction.

[0061] As used herein, the term "cerebral infarction" refers to a disease in which ischemic damage occurs due to occlusion of a cerebral blood vessel, resulting in neurological dysfunction. Ischemic brain damage caused by cerebral infarction, like the hematoma formation caused by intracerebral hemorrhage described above, leads to an excessive inflammatory response, ultimately becoming a major cause of cerebral edema. Therefore, "cerebral edema" may also be included among the inflammatory or autoimmune diseases that can be prevented or treated with the composition of the present invention.

[0062] The term "cerebral edema" in this specification refers to a disease in which brain tissue enlarges due to the influx of fluid into cells or tissues caused by any brain tissue damage, such as trauma, tumor, or inflammation. Cerebral edema occurs as a result of an inflammatory response within the brain tissue, and when extensive cerebral edema occurs within a confined intracranial space, physical pressure is applied to the surrounding tissue. In severe cases, brain herniation, brainstem compression, and hydrocephalus may occur, which may not only cause additional neurological damage but also lead to death.

[0063] According to a specific embodiment of the present invention, the cerebral edema to be prevented or treated in the present invention is cerebral edema caused by stroke, cerebral infarction, intracranial hemorrhage, brain neoplasm, traumatic brain injury, encephalitis, inflammatory brain disease, demyelinating brain disease, or intracranial vasculitis. More specifically, the cerebral edema is cerebral edema caused by cerebral infarction.

[0064]

[0065] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is dermatitis.

[0066] As used herein, the term “dermatitis” refers to any disease in which skin tissues, including the epidermis, dermis, and subcutaneous tissues, are damaged and the inherent biological functions of skin tissues, including the barrier function, are impaired due to excessive or unwanted immune responses or inflammation caused thereby. The dermatitis that can be prevented or treated with the composition of the present invention may be selected from the group consisting of atopic dermatitis, eczema, erythema multiforme, erythema nodosum, and pyoderma gangrenosum, and most specifically, may be atopic dermatitis.

[0067] The term "atopic dermatitis" used herein refers to a chronic allergic inflammatory skin disease that occurs in skin tissue. It is common in children and can persist into adulthood. Atopic dermatitis is influenced by a family history of allergic diseases and is triggered by allergens such as food or dust mites. Most currently used medications, including steroids, are symptomatic and fail to address the underlying cause, making it a representative intractable skin disease.

[0068]

[0069] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is traumatic brain injury.

[0070] The term “traumatic brain injury” as used herein refers to a disease in which brain tissue is damaged by external physical force and brain function, including cognitive function, deteriorates. Oxidative stress caused by excessive generation of reactive oxygen species after trauma is a major secondary damage mechanism of traumatic brain injury, and reducing reactive oxygen species explosively generated immediately after trauma is very important in preventing additional tissue damage. In addition, just like hematoma formation due to intracerebral hemorrhage and ischemic brain damage due to cerebral infarction described above, traumatic brain injury is also one of the major causes of cerebral edema. Therefore, prevention or treatment of traumatic brain injury using the composition of the present invention can lead to prevention or treatment of “cerebral edema.”

[0071]

[0072] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is a gunshot wound.

[0073] The term “gunshot injury” as used herein refers to damage caused by the insertion or penetration of body tissue by a projectile such as a bullet fired from a firearm, and is used herein with the same meaning as “inflammation due to gunshot wound” or “sepsis due to gunshot wound.” Injuries due to gunshot wounds include bleeding, fractures, organ damage, wound infection, inflammation, and death. Long-term complications include intestinal obstruction, growth retardation, neurogenic bladder and paralysis, recurrent respiratory distress and pneumothorax, and early dementia due to hypoxic brain damage. In particular, a penetrating bullet forms a perforation in the tissue, causing bleeding and the influx of foreign substances, leading to inflammation and sepsis. Therefore, controlling the inflammatory response and reactive oxygen species that rapidly increase immediately after gunshot wound are important factors in determining the patient’s survival, along with surgical suturing.

[0074] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is acute respiratory distress syndrome.

[0075] In this specification, the term “acute respiratory distress syndrome (ARDS)” refers to a pathological condition in which respiratory failure is caused by a rapid decline in lung function, resulting in a decrease in the partial pressure of oxygen in the arterial blood and an accumulation of carbon dioxide. It is also called “acute lung injury (ALI).” Representative causes of rapid decline in lung function include sepsis caused by systemic infection, trauma, massive blood transfusion, and pancreatitis. In addition, the massive influx of foreign substances due to bacteria or trauma causes severe inflammation in the lungs, leading to a rapid loss of lung function. ARDS is classified into acute, proliferative, and fibrotic phases according to its course. In the acute phase, excessive inflammatory response is the main factor, so controlling inflammation is of utmost importance in blocking the progression of the disease.

[0076]

[0077] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is inflammatory bowel disease (IBD).

[0078] The term "inflammatory bowel disease (IBD)" as used herein encompasses inflammatory disorders occurring in the gastrointestinal tract, including, but not limited to, ulcerative colitis and Crohn's disease, the most common forms. IBD is a chronic and intractable disease characterized by symptoms such as abdominal pain, diarrhea, bloody stool, and weight loss that persist for several months or longer. Fundamental treatment is only possible by controlling the inflammation occurring throughout the digestive tract and the resulting multiple oxidative stresses.

[0079]

[0080] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is subarachnoid hemorrhage (SAH).

[0081] The term “subarachnoid hemorrhage (SAH)” in this specification refers to a disease in which blood leaks from the cerebral blood vessels into the subarachnoid space, causing various inflammatory responses and brain cell damage in the brain tissues located in the periphery of the brain parenchyma. Because SAH occurs in the subarachnoid space, which surrounds the entire brain, it causes extensive damage throughout the brain, which can lead to immediate death or serious permanent brain damage. Brain damage caused by SAH is caused by a chain reaction of blood components such as hemoglobin and thrombin that initially induce inflammation in the brain tissue and cause oxidative damage through reactive oxygen species. Therefore, it is important to minimize neurological damage by quickly controlling reactive oxygen species and blocking the inflammatory response.

[0082]

[0083] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is sepsis.

[0084] The term "sepsis" in this specification encompasses a harmful systemic inflammatory response to external infectious agents such as viruses and bacteria, and the resulting pathological condition, including complications such as "severe sepsis" and "septic shock." Sepsis is accompanied by fever, dyspnea, and rapid changes in white blood cell count, leading to multiple organ failure and ultimately death at a high rate. Simultaneous measures are taken to alleviate septic shock, including administering antibiotics and antifungal agents to eliminate the source of infection, as well as providing fluids, maintaining blood pressure, transfusions, and correcting acidosis. However, fundamental treatment is only possible by removing the excessive reactive oxygen species caused by the inflammatory response.

[0085]

[0086] According to a specific embodiment of the present invention, the inflammatory or autoimmune disease is inflammatory liver disease.

[0087] The term “inflammatory liver disease” in this specification encompasses a series of diseases involving damage to liver tissue caused directly or indirectly by excessive or unwanted immune responses or inflammatory reactions induced by various causes.

[0088] More specifically, the inflammatory liver disease is selected from the group consisting of viral hepatitis, toxoplasma hepatitis, alcoholic liver disease, toxic liver disease, acute and subacute hepatic failure, liver abscess, nonspecific reactive hepatitis, liver infarction, hepatic veno-occlusive disease, injury of the liver or gallbladder, and liver transplantation-related hepatitis.

[0089]

[0090] When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is one commonly used in formulation, and includes, but is not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0091] The pharmaceutical composition of the present invention can be administered through various routes of administration, specifically parenterally, more specifically parenterally, and even more specifically intravenously, intraarterially, subcutaneously, intraperitoneally, intradermally, intramuscularly, intracerebroventricularly, intrathecally, by inhalation, intranasally, intraarticularly, or topically.

[0092] The appropriate dosage of the pharmaceutical composition of the present invention may be prescribed in various ways depending on factors such as the formulation method, administration method, patient age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity. The preferred dosage of the pharmaceutical composition of the present invention is within the range of 0.0001-1000 mg / kg for adults.

[0093] The pharmaceutical composition of the present invention can be manufactured in a unit dose form or can be manufactured by inserting it into a multi-dose container by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily performed by a person having ordinary skill in the art to which the present invention pertains, and the like. In this case, the formulation may be in the form of a solution, suspension, syrup or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersing agent or stabilizer.

[0094] According to another aspect of the present invention, there is provided a method for preventing or treating an inflammatory or autoimmune disease, comprising administering to a subject the cerium oxide nanocomposite of the present invention or a pharmaceutical composition comprising the same as an active ingredient. The cerium oxide nanocomposite used in the present invention and the inflammatory or autoimmune disease that can be prevented or treated therewith have already been described above, and therefore, to avoid excessive repetition, a detailed description thereof will be omitted.

[0095] According to another aspect of the present invention, the present invention provides a method for producing a cerium oxide nanocomposite, comprising the following steps:

[0096] (a) A step of preparing a mixed solution by adding a cerium precursor, a polymer represented by the following chemical formula 1, and a crosslinking compound represented by the following chemical formula 2 to a C1 to C3 alcohol solvent;

[0097] Chemical Formula 1

[0098]

[0099] In the above chemical formula 1, R1 and R2 are each independently hydrogen or oxygen, represents a single bond or a double bond, l is 1 or 2, and m is an integer from 100 to 1000;

[0100] Chemical Formula 2

[0101]

[0102] In the above chemical formula 2, n is an integer from 3 to 7;

[0103] (b) a step of sequentially heating and cooling the mixed solution to obtain cerium oxide nanoparticles; and

[0104] (c) A step of washing the above cerium oxide nanoparticles with an inorganic salt solution.

[0105] According to the present invention, by performing the step (a), a core-shell structure can be formed in which a compound of formula 1 is bonded to a cerium oxide nanoparticle core by using a compound of formula 2 as a crosslinker, thereby forming a modified outer layer.

[0106] In this specification, the term “coating” means forming a new layer of a certain thickness by applying a specific material on the target surface, and the target surface and the modified layer material can be modified through ionic bonds or non-covalent bonds. The term “non-covalent bond” is a concept that includes not only physical bonds such as adsorption, cohesion, entanglement, and entrapment, but also bonds that occur when interactions such as hydrogen bonds and van der Waals bonds act alone or in combination with the above physical bonds. In the present invention, when the compound of chemical formula 1 is modified on the surface of the cerium oxide nanoparticles, it may form a closed layer that completely surrounds the target surface to be modified, or it may form a partially closed layer.

[0107] According to a specific embodiment of the present invention, the C1 to C3 alcohol solvent is ethanol.

[0108] According to a specific embodiment of the present invention, the cerium precursor is at least one precursor selected from the group consisting of cerium(III) acetate hydrate, cerium(III) acetylacetonate hydrate, cerium(III) carbonate hydrate, cerium(III) fluoride, cerium(III) chloride, cerium(III) chloride heptahydrate, cerium(III) bromide, cerium(III) iodide, cerium(III) nitrate hexahydrate, cerium(III) oxalate hydrate, cerium(III) sulfate, and cerium(III) sulfate hydrate. More specifically, it is cerium(III) nitrate hexahydrate.

[0109]

[0110] According to a specific embodiment of the present invention, the heating is performed at 60 to 80°C. More specifically, it is performed at 65 to 75°C, and most specifically, it is performed at about 70°C.

[0111] According to a specific embodiment of the present invention, the cooling is performed by reducing the temperature to 35 to 50°C. More specifically, it is performed by reducing the temperature to 40 to 50°C, and most specifically, it is performed by reducing the temperature to about 45°C.

[0112] In this specification, the term “washing” means a process of physically and chemically removing impurities bound to a reaction product using a washing solution or buffer solution in order to reduce the content of unwanted impurities in the process of separating and purifying a reaction product and increase the purity of the final product to be obtained.

[0113] As used herein, the term "inorganic salt" refers to a salt derived from an inorganic substance that is a combination of cations and anions through ionic bonds in an aqueous solution and does not contain a CH bond. According to a specific embodiment of the present invention, the inorganic salt is selected from the group consisting of NaCl, CaCl2, KCl, MgCl2, and combinations thereof.

[0114] More specifically, the inorganic salt solution used in the present invention is a NaCl solution.

[0115] According to a specific embodiment of the present invention, the NaCl solution has a concentration of 0.15 to 0.35 M.

[0116] The present inventors have discovered that, simply by adding a washing step with a sodium chloride solution having a concentration of 0.15 to 0.35 M after the completion of the reaction, particles having an optimal uniform diameter can be efficiently obtained while completely removing unreacted residues and toxicity due to nitrates. More specifically, the NaCl solution of the present invention has a concentration of 0.20 to 0.30 M, and most specifically, a concentration of about 0.25 M.

[0117]

[0118] The features and advantages of the present invention are summarized as follows:

[0119] (a) The present invention provides a cerium oxide nanocomposite, a method for producing the same, and a composition for preventing or treating inflammatory or autoimmune diseases comprising the same as an active ingredient.

[0120] (b) The present invention can be used as an excellent therapeutic composition in which both biostability and efficiency in removing reactive oxygen species in the body are maximized by modifying the surface of cerium oxide nanoparticles with a pyrrolidone derivative polymer in an optimal amount.

[0121] (c) The method for manufacturing a nanocomposite of the present invention can form uniform particles having an optimal diameter while completely removing reaction residues and toxicity due to nitrates with only a simple process of washing with a sodium chloride solution of a specific concentration.

[0122] (d) The present invention can be usefully applied as an excellent therapeutic composition that significantly suppresses inflammatory responses in various tissues and, in particular, minimizes damage to nerve cells caused by excessive inflammatory responses around a hematoma due to intracerebral hemorrhage, thereby restoring neurological function and significantly improving the survival rate of patients.

[0123]

[0124] Figure 1 is a diagram showing a schematic diagram (Figure 1a) and a transmission electron microscope observation result (Figure 1b) of the cerium oxide nanocomposite of the present invention, respectively.

[0125] FIG. 2 is a drawing showing the results of analyzing the changes in particle size after synthesis and dispersion stability and surface charge in a biomimetic environment according to the content of PVP (polyvinyl pyrrolidone), a polymer modified on the outer shell of the cerium oxide nanocomposite of the present invention (FIG. 2a) and the results of converting the content of PVP in the dried cerium oxide nanocomposite of the present invention into a unit of cerium (FIG. 2b).

[0126] Figure 3 is a drawing showing the results of measuring the particle size of the cerium oxide nanocomposite of the present invention over time by exposing it to a biomimetic environment using a dynamic light scattering device.

[0127] Figure 4 shows the results of comparing the surface charge of a conventionally developed cerium oxide nanocomposite (CX213) and the cerium oxide nanocomposite of the present invention.

[0128] Figure 5 shows the results of observing the reaction residue (Figure 5a) and particle size change (Figure 5b) according to the concentration of the sodium chloride aqueous solution used in the purification process of the cerium oxide nanocomposite of the present invention.

[0129] Figure 6 shows the results of a comparative analysis of the therapeutic effects of a conventionally developed cerium oxide nanocomposite (CX213) and the cerium oxide nanocomposite of the present invention (CX301) on severe cerebral infarction through observation of survival rates after intravenous injection into a severe cerebral infarction (MIS) rat model.

[0130] Figure 7 shows the results of a comparative analysis of the therapeutic effects of a conventionally developed cerium oxide nanocomposite (CX213) and the cerium oxide nanocomposite of the present invention (CX301) on severe cerebral infarction through infarct volume.

[0131] Figure 8 shows the results of comparative analysis of brain water content due to cerebral edema to evaluate the intracerebral hemorrhage treatment effect of the cerium oxide nanocomposite of the present invention.

[0132] FIG. 9 is a drawing showing the results of measuring the cylinder asymmetry score in an artificial traumatic brain injury model to evaluate the treatment effect of the cerium oxide nanocomposite of the present invention on traumatic brain injury.

[0133] Figure 10 is a drawing showing the results of measuring the degree of water loss (Figure 10a) and the degree of epidermal thickening and inflammatory mast cell infiltration (Figure 10b) in a DNCB (2,4-dinitro-chlorobenzene)-induced atopic dermatitis animal model to evaluate the dermatitis treatment effect of the cerium oxide nanocomposite of the present invention.

[0134] Figure 11 is a drawing showing the results of measuring body weight change and disease activity score (DAI) in a DSS (dextran sodium sulfate)-induced inflammatory bowel disease animal model to evaluate the inflammatory bowel disease treatment effect of the cerium oxide nanocomposite of the present invention.

[0135] Figure 12 is a drawing showing the results of measuring the change in survival rate according to treatment with the cerium oxide nanocomposite of the present invention in a CLP-induced sepsis animal model to evaluate the sepsis treatment effect.

[0136] Figure 13 is a drawing showing the results of evaluating the survival rate of hepatocytes (hepa-1c1c7) induced with toxic stimulation by pyrogallol by treating them with the cerium oxide nanocomposite of the present invention to confirm the treatment effect for inflammatory liver disease.

[0137] Figure 14 is a drawing showing the results of measuring the change in survival rate according to treatment with the cerium oxide nanocomposite of the present invention in an animal model in which subarachnoid hemorrhage was induced to evaluate the therapeutic effect of subarachnoid hemorrhage.

[0138]

[0139] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples, in accordance with the gist of the present invention.

[0140]

[0141] Example

[0142] Example 1: Synthesis of cerium oxide nanocomposites

[0143] A first solution was prepared by dissolving 6-aminohexanoic acid (37.75 g, Sigma-Aldrich, St. Louis, MO) in deionized water (1.7 L). A second solution was prepared by adding ethyl alcohol (1.4 L) to the first solution while stirring, and weighing and adding 132.83 g of polyvinylpyrrolidone (PVP, Ashland), followed by heating to 70°C in air. Meanwhile, a third solution was prepared by dissolving cerium(III) nitrate hexahydrate (Ce(NO3)3·6H2O, 31.05 g, Alfa Aeser, Ward Hill, MA) in ethyl alcohol (2.9 L) at room temperature (approximately 20°C). Thereafter, the third solution was added to the second solution to prepare a fourth solution. The temperature of the fourth solution was maintained at 70°C for 2 hours and then lowered to 45°C. Through this process, cerium oxide nanoparticles with 6-aminohexanoic acid and polyvinylpyrrolidone bound to their surfaces were obtained (Figures 1a and 1b). Subsequently, the nanoparticles were washed five times with a mixture of acetone and aqueous sodium chloride solution to remove unreacted substances.

[0144]

[0145] Example 2: Exploration of the optimal content of polyvinyl pyrrolidone used as an outer layer polymer

[0146] In order to explore the optimal content of the polyvinyl pyrrolidone dispersion stabilizer forming the outer layer of the cerium oxide nanoparticles of the present invention, the particle formation tendency according to the PVP content and the stability of dispersion in a biomimetic environment were analyzed. During the synthesis of the cerium oxide nanocomposite of Example 1, the content ratio of PVP was adjusted from 15 to 30 mg PVP / total batch mL, and the particle size of the synthesized product and the dispersion stability (Z-Average, nm) and surface charge (Zeta-potential, mV) in a biomimetic environment containing protein and sodium bicarbonate were measured by dynamic light scattering analysis. As a result, it was found that when PVP was applied at a content ratio of 20 to 25 mg PVP / total batch mL, the optimized particle characteristics were exhibited (Fig. 2a).

[0147] Additionally, in order to more clearly measure the optimal PVP content range derived above based on the final product, the inventors of the present invention lyophilized the entire synthesized nanocomposite and then converted the PVP content in the dried nanocomposite into units of cerium. As a result, it was confirmed that the PVP and cerium oxide nanoparticles in the final nanocomposite exhibited the best particle characteristics when the content ratio was 7.5 to 10.9 (7.5 - 10.9 mg PVP / mg Ce), and the optimal content ratio was 8.7 (Fig. 2b). This composition ratio explored in the present invention can be said to be the result of exploring the most suitable conditions when considering the effect of the size of the nanocomposite on dispersion, function, and biological stability.

[0148]

[0149] Experimental Example 1: Comparison of biostability and dispersibility between previously developed particles and the particles of the present invention in a biomimetic environment.

[0150] In order to compare the biostability (dispersibility) of the ceria nanocomposite (CX213) previously developed by the present inventors, in which PGA (polyglutamic acid) is modified in the outermost layer as an additional dispersion stabilizer, and the cerium oxide nanoparticles (CX301) of the present invention, in which the introduction of the outermost polymer layer is omitted, in a biomimetic environment, the change in particle size was analyzed over time in 0.05 M PBS (phosphate buffer saline). Samples were collected every 0, 20, 40, and 60 minutes, respectively, and the particle size was analyzed using a dynamic light scattering device. As a result, it was found that the CX213 nanocomposite showed particle aggregation reaching several hundred nm as the reaction time elapsed, whereas the CX301 of the present invention maintained a constant particle size even over time (Fig. 3).

[0151]

[0152] Experimental Example 2: Surface charge analysis between previously developed particles and particles of the present invention.

[0153] The inventors of the present invention measured the surface charge of CX213, which was developed previously, and CX301, to evaluate the difference in biostability and dispersibility depending on the presence or absence of PVP optimization. The synthesized cerium oxide nanocomposites were dispersed in deionized water, and the surface characteristics of the ceria nanocomposites were analyzed using a dynamic light scattering device. As a result, the surface charge was -17.5 mV for CX213 and 0 mV for CX301, which showed a clear difference between the two particles (Fig. 4). Through this, it was found that the present invention can minimize interactions in the biological environment compared to CX213, which has a strong negative charge due to the inclusion of the outermost PGA polymer, as the surface charge converges to 0 mV.

[0154]

[0155] Experimental Example 3: Evaluation of the Effect of Addition of Sodium Chloride Solution During Purification on Reaction Residue Concentration and Particle Size

[0156] In order to measure the particle size of the reaction residue and the final product of the cerium oxide nanocomposite obtained by adding sodium chloride solution of different concentrations during the purification process, the concentration of the sodium chloride aqueous solution was adjusted to 0, 0.25, 0.5, 1.0, 2.5, and 5 M during the manufacturing process of the cerium oxide nanocomposite of Example 1, and the residual content of the reaction residue, especially nitrate, was measured using a nitrate indicator pad PP strip, Johnson. In addition, the particle size was confirmed using a dynamic light scattering device. As a result, as shown in Fig. 5a, it was found that the use of 0.25 M sodium chloride aqueous solution during the purification process was the most effective in removing the reaction residue, especially nitrate. Through this, it was found that the manufacturing process of the present invention is efficient in both reducing residue in pharmaceuticals and removing toxicity caused by nitrate. In addition, the particle size was also most optimized (20 nm) when using a 0.25 M sodium chloride aqueous solution, confirming that the addition of sodium chloride had a significant effect on the formation stability of the particles (Fig. 5b).

[0157]

[0158] Experimental Example 4: Evaluation of the Treatment Efficacy of Non-Infectious Inflammatory Diseases through Survival Rates

[0159] In order to compare and evaluate the non-infectious inflammatory disease treatment effect of CX213, a conventionally developed cerium oxide nanocomposite, and CX301 of the present invention manufactured in Example 1, SD (Sparague-Dawley) rats (Coatech Co., Ltd.) were anesthetized with isoflurane, and then the left middle cerebral artery was ligated with a 4-0 proline suture to induce severe cerebral infarction (MIS), a representative non-infectious inflammatory disease. One hour after inducing severe cerebral infarction, the two types of cerium oxide nanocomposites were intravenously injected once over 5 minutes at 0.5 mg Ce / kg, and an equal volume of saline was injected as a control. The SD rats were periodically checked for death until 7 days after inducing MIS, and the survival rate was measured. The experiment was conducted on 18 and 15 SD rats in Example 1 and the control group, respectively, and the average value was expressed as the survival rate. As shown in Fig. 6, the CX213 nanocomposite showed a survival rate increase effect of 2.25 times compared to the control group (control group: 28.57%, CX213 administration group: 64.28%), whereas the group injected with CX301 of the present invention showed a survival rate increase effect of up to 6 times (control group: 13.33%, CX301 administration group: 80.00%).

[0160] Through this, it was observed that CX301, a novel cerium oxide nanocomposite of the present invention, has a therapeutic effect 2.6 times greater than CX213, which the inventors previously developed, thereby confirming that it can be usefully utilized as an effective therapeutic composition for non-infectious inflammatory diseases.

[0161]

[0162] Experimental Example 5: Evaluation of the therapeutic effect of non-infectious inflammatory diseases through infarct volume.

[0163] Brain tissues were collected from the SD rats of Experimental Example 5 above, coronal sections were obtained, and Nissl staining was performed to confirm the degree of infarction. After staining the tissues, the volume was calculated based on the area of ​​each section, and as a result, the infarction volume was confirmed to be reduced by 18.58% in the CX213 administration group and 51.22% in the CX301 administration group of the present invention compared to the control group. Accordingly, the cerium oxide nanocomposite of the present invention showed a therapeutic effect that was as much as 2.76 times higher in terms of infarction volume than the nanocomposite developed previously (Fig. 7). Through this, it was confirmed from various aspects that the CX301 nanocomposite of the present invention not only significantly increases the survival rate of the disease model but also greatly reduces the infarction volume of actual brain tissue, exhibiting a remarkably excellent therapeutic effect.

[0164]

[0165] Experimental Example 6: Evaluation of the therapeutic effect on additional non-infectious inflammatory diseases.

[0166] intracerebral hemorrhage

[0167] Intracerebral hemorrhage is a disease in which persistent neuronal damage occurs due to an excessive inflammatory response around the hematoma. This study aimed to determine whether the anti-inflammatory action of the cerium oxide nanocomposite of the present invention can alleviate the symptoms of intracerebral hemorrhage. First, randomly assigned SD rats were anesthetized with 2.5% isoflurane and the rats' heads were fixed in a stereotaxic frame. Then, a cranial perforation was performed at a location 0.02 mm posterior to the bregma and 0.30 mm lateral to the midline in the coronal plane. To induce intracerebral hemorrhage, 1 μl of 0.5 unit of collagenase VII was injected to a depth of 0.66 mm, and then 0.5 mg / kg of nanocerium complex or saline was intravenously injected twice, 1 hour and 24 hours after model induction. To assess the severity of the disease, the degree of cerebral edema was determined by measuring brain water content on the third day. The results of the brain water content measurement showed a 22.72% improvement in the cerium oxide nanocomposite treatment group compared to the saline-treated control group (Fig. 8). These results confirmed that the cerium oxide nanocomposite of the present invention exhibits excellent therapeutic effects against intracerebral hemorrhage.

[0168]

[0169] traumatic brain injury

[0170] Traumatic brain injury (TBI) is a disease in which secondary neuronal damage occurs due to an inflammatory response caused by excessive generation of reactive oxygen species. The present inventors sought to verify the therapeutic effect of the cerium oxide nanocomposite of the present invention on TBI. To induce traumatic brain injury, SD rats were anesthetized with 2.5% isoflurane and their heads were fixed in a stereotaxic frame. Then, a skull perforation was performed at a location 3.5 mm anterior to the bregma in the coronal plane and 3.5 mm lateral to the midline. To inflict brain damage at the designated site, a pinpoint precision impactor (PCI3000, Hatteras) was equipped with an impactor tip with a diameter of 5 mm, and an impact depth of 6 mm was set to deliver impact at a velocity of 4 m / s. One hour after inducing the disease, a single intravenous dose of 0.1 mg / kg of the cerium oxide nanocomposite of the present invention was administered, and the degree of motor dysfunction was determined through a cylinder test. For accurate measurement, a mirror was installed on the opposite side of the behavioral filming to avoid blind spots, and then the animals were placed in the cylinder and their movements were observed for 10 minutes. The measurements were recorded after confirming the feet used for touching the wall, finding balance, and landing, and the scores were evaluated using the formula [(I+1 / 2B) / (I+C+B)]x100 (brain damage area (ipsilateral, I), contralateral area (contralateral, C), and bilateral area (bilateral, B)). As a result of the test, a 47.02% improvement in motor ability was observed in the group administered the cerium oxide nanocomposite of the present invention compared to the group administered saline on the third day of measurement (Fig. 9). Through these results, it was confirmed that the cerium oxide nanocomposite of the present invention can be usefully applied as an effective therapeutic composition for traumatic brain injury, which is an intractable disease.

[0171]

[0172] dermatitis

[0173] In inflammatory skin diseases caused by various reasons, excessive inflammatory response due to reactive oxygen species is a cause of worsening the severity of dermatitis. In order to confirm whether the cerium oxide nanocomposite of the present invention can also alleviate the severity of inflammatory skin diseases, a dermatitis model was induced using hairless SKH-1 mice and the efficacy of the cerium oxide nanocomposite was verified. To induce atopic dermatitis, 0.2 ml of 1% 2,4-dinitro-chlorobenzene (DNCB) was applied to the backs of mice daily for 35 days, and from the 17th day of application, 0.5 mg / kg or 1 mg / kg of the nanocomposite of the present invention was applied to the same area at 0.2 ml each. In order to observe the dermatitis improvement effect, the degree of skin moisture loss was measured once every three days using a moisture measuring device (GP skin barrier, GP skin). On the final day, skin tissue was obtained and stained to confirm changes in epidermal thickness and the degree of inflammatory cell infiltration. As a result of the experiment, as of the final end date, a water loss improvement effect of 12.13% was observed in the 0.5 mg / kg cerium oxide nanocomposite treatment group and 19.19% in the 1 mg / kg treatment group (Fig. 10a). It was also confirmed that the degree of thickening was alleviated by 38.78% and 47.09% in the 0.5 mg / kg or 1 mg / kg cerium oxide nanocomposite treatment group, respectively (Fig. 10b, left). It was confirmed that the degree of inflammatory mast cell infiltration, which is characteristically observed in the dermatitis model, was reduced by 27.21% and 33.05%, respectively (Fig. 10b, right).

[0174]

[0175] colitis

[0176] Oxidative stress plays an essential role in the pathogenesis and progression of inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn's disease (CD). This study aimed to verify whether the antioxidant and anti-inflammatory activities of the cerium oxide nanocomposite of the present invention can also suppress the symptoms and progression of inflammatory bowel disease. To establish a disease model, C57BL / 6 mice were administered either regular drinking water or 2% dextran sodium sulfate (DSS), a substance that induces epithelial cell damage, daily for 10 days. The cerium oxide nanocomposite was administered 1 mg / kg 0.2 ml intrarectally daily from 7 days before the induction of inflammatory bowel disease with DSS until the end of the test period. In order to check the severity of the disease, daily body weight, stool condition, and rectal bleeding were recorded, and 0 points were given for normal, 1 point for weight loss of less than 10%, 2 points for weight loss of less than 15% and loose stool, 3 points for weight loss of less than 20% and 4 points for weight loss of more than 20% and diarrhea or bloody stool, and the severity was evaluated by calculating the DAI (disease activity score). As a result of the experiment, it was observed that in the group administered with cerium oxide nanocomposite, compared to the control group administered with physiological saline, body weight loss was suppressed by 8.05% (left side of Fig. 11), and disease severity was also alleviated by 24.21% (right side of Fig. 11). Through this, it was found that the cerium oxide nanocomposite of the present invention can be used as an effective therapeutic composition for inflammatory bowel disease while exhibiting significant antioxidant and anti-inflammatory effects even under conditions of direct administration to the intestines.

[0177]

[0178] blood poisoning

[0179] Sepsis is a disease that causes tissue damage and organ failure due to the body's excessive response to various infections. The inventors of the present invention attempted to verify the sepsis treatment effect of the cerium oxide nanocomposite of the present invention. Cecal Ligation and Puncture (CLP) was used to induce sepsis. Specifically, SD rats were anesthetized with 2.5% isoflurane, and the cecum of the rats was ligated and punctured to induce intraperitoneal infection. One hour after the disease induction, the cerium oxide nanocomposite of the present invention was administered intravenously as a single dose at a cerium concentration of 0.5 mg / kg. The survival rate for 6 days was measured, and the experiment was performed on 19 SD rats in each of the experimental and control groups, and the survival rate was expressed as the survival rate. As a result, a survival rate improvement effect approximately three times higher was observed in the cerium oxide nanocomposite treatment group of the present invention compared to the saline-treated control group on the 6th day (control group: 5.3%, experimental group: 15.8%) (Fig. 12). This confirmed that the cerium oxide nanocomposite of the present invention also has an excellent therapeutic effect on sepsis.

[0180] inflammatory liver disease

[0181] Inflammatory liver disease is a disease in which the liver cannot perform normal functions due to liver damage, metabolic disorders, and accumulation of toxic substances. The inventors of the present invention sought to verify the therapeutic effect of the cerium oxide nanocomposite of the present invention on inflammatory liver disease. To induce the disease, the Hepa-1c1c7 hepatocyte cell line was treated with 500 μM of pyrogallol, a cytotoxic substance, to induce acute inflammatory liver disease, and simultaneously treated with the cerium oxide nanocomposite of the present invention at a concentration of 0.01 to 5 μM. After 3 hours of treatment, the viability of Hepa-1c1c7 cells was measured through the NRU assay, a biochemical analysis method that measures the uptake of neutral red dye into living cells to evaluate cell viability. As a result, compared to the control group that was not treated with pyrogallol, the cells treated with pyrogallol showed a survival rate of about 50.43%, whereas when treated together with the cerium oxide nanocomposite of the present invention, the cell survival rate was 74.55% at a minimum of 0.01 μM and about 93.19% at a maximum of 5 μM, showing an increase in survival rate of at least 47.81% and at most 84.78% compared to the cells treated with pyrogallol alone (Fig. 13). These results show that the cerium oxide nanocomposite of the present invention can be used as an effective therapeutic composition for inflammatory liver disease.

[0182]

[0183] subarachnoid hemorrhage

[0184] Subarachnoid hemorrhage (SAH) is a disease in which blood leaks into the subarachnoid space of the brain, mainly caused by a ruptured cerebral aneurysm. The inventors of the present invention sought to verify the therapeutic effect of the cerium oxide nanocomposite of the present invention on subarachnoid hemorrhage. After anesthetizing SD rats with 2.5% isoflurane, the heads of the rats were fixed, an incision was made at the base of the neck to find the common carotid artery, internal carotid artery, and external carotid artery, and a filament was inserted into the external carotid artery, extending the filament to the middle cerebral artery. After that, the middle cerebral artery was perforated to induce bleeding, thereby inducing subarachnoid hemorrhage. One hour after the induction, the cerium oxide nanocomposite of the present invention was administered intravenously once to the rats at a concentration of 0.5 mg / kg based on the cerium concentration. The survival rate for 10 days was measured, and the survival rate was measured and presented for five rats each in the control group and the experimental group. As a result, a survival rate increase effect twice as high was observed in the cerium oxide nanocomposite treatment group of the present invention compared to the control group treated with saline solution on the 10th day (control group: 40%, experimental group: 80%) (Fig. 14a). In addition, as a result of measuring the water content in the brain of the control and experimental groups on the 3rd day, the water content on the contralesional side was 79.5% in the control group and 79.8% in the experimental group, showing a difference of less than 0.3%, whereas the water content on the ipsilesional side was 80.8% in the control group and 80.1% in the experimental group, showing a water content reduction effect of 0.7% (Fig. 14b). Through this, it was confirmed that the cerium oxide nanocomposite of the present invention also exhibits an excellent therapeutic effect on subarachnoid hemorrhage.

[0185]

[0186] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred embodiments and do not limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. Cerium oxide nanocomposite comprising: (a) a core layer of cerium oxide nanoparticles; and (b) an outer layer comprising a polymer represented by the following chemical formula 1: Chemical Formula 1 In the above chemical formula, R1 and R2 are each independently hydrogen or oxygen, represents a single bond or a double bond, l is 1 or 2, and m is an integer from 100 to 1000.

2. A nanocomposite according to claim 1, characterized in that the cerium oxide nanoparticles are selected from the group consisting of cerium(III) oxide (Ce2O3) nanoparticles, cerium(IV) oxide (CeO2) nanoparticles, and mixtures thereof.

3. A nanocomposite characterized in that in the first paragraph, R1 is hydrogen, R2 is oxygen, and l is 1 in the chemical formula 1.

4. A nanocomposite according to claim 3, characterized in that the polymer represented by the chemical formula 1 and the cerium oxide nanoparticles have a content ratio of 7:1 to 11:

1.

5. In the first paragraph, the nanocomposite is characterized in that it additionally includes a multifunctional ligand represented by the following chemical formula 2: Chemical Formula 2 In the above chemical formula, n is an integer from 3 to 7.

6. A nanocomposite according to claim 5, wherein n in the chemical formula 2 is 5.

7. A nanocomposite according to claim 1, characterized in that the nanocomposite has an average particle diameter of 5 nm to 80 nm.

8. A nanocomposite according to claim 1, characterized in that the nanocomposite has a surface charge (Zeta potential, mV) of -1.0 to 1.0 mV.

9. A composition for preventing or treating inflammatory or autoimmune diseases, comprising the nanocomposite of any one of claims 1 to 8 as an active ingredient. 10.In claim 9, the inflammatory or autoimmune disease is intracerebral hemorrhage (ICH), cerebral infarction, dermatitis, traumatic brain injury, inflammatory bowel disease (IBD), subarachnoid hemorrhage (SAH), cerebral edema, hypoxic ischemic encephalopathy, stroke, traumatic spinal cord injury, gunshot injury, acute respiratory distress syndrome, cytokine storm syndrome, sepsis, systemic inflammatory response syndrome, peritonitis, multiple sclerosis, rheumatoid arthritis, reactive arthritis, type 1 diabetes, systemic A composition characterized in that the disease is at least one disease selected from the group consisting of lupus erythematosus, idiopathic pulmonary fibrosis, polymyositis, dermatomyositis, localized scleroderma, systemic scleroderma, Sjogren's syndrome, Raynaud's phenomenon, Bechet's disease, Kawasaki's disease, primary biliary sclerosis, primary sclerosing cholangitis, psoriasis, myasthenia gravis, autoimmune vasculitis, primary angiitis of the central nervous system, and inflammatory liver disease.

11. A composition according to claim 10, wherein the inflammatory or autoimmune disease is intracerebral hemorrhage (ICH).

12. A composition according to claim 10, wherein the inflammatory or autoimmune disease is cerebral infarction.

13. A composition according to claim 10, wherein the inflammatory or autoimmune disease is dermatitis.

14. A composition according to claim 10, wherein the inflammatory or autoimmune disease is traumatic brain injury.

15. A composition according to claim 10, wherein the inflammatory or autoimmune disease is inflammatory bowel disease (IBD).

16. A composition according to claim 10, wherein the inflammatory or autoimmune disease is subarachnoid hemorrhage (SAH).

17. A composition according to claim 10, wherein the inflammatory or autoimmune disease is sepsis.

18. A composition according to claim 10, wherein the inflammatory or autoimmune disease is inflammatory liver disease.

19. A method for producing a cerium oxide nanocomposite comprising the following steps: (a) A step of preparing a mixed solution by adding a cerium precursor, a polymer represented by the following chemical formula 1, and a crosslinking compound represented by the following chemical formula 2 to a C1 to C3 alcohol solvent; Chemical Formula 1 In the above chemical formula 1, R1 and R2 are each independently hydrogen or oxygen, represents a single bond or a double bond, l is 1 or 2, and m is an integer from 100 to 1000; Chemical Formula 2 In the above chemical formula 2, n is an integer from 3 to 7; (b) a step of sequentially heating and cooling the mixed solution to obtain cerium oxide nanoparticles; and (c) A step of washing the above cerium oxide nanoparticles with an inorganic salt solution.

20. A method according to claim 19, characterized in that the C1 to C3 alcohol solvent is ethanol.

21. A method according to claim 19, wherein the cerium precursor is at least one precursor selected from the group consisting of cerium(III) acetate hydrate, cerium(III) acetylacetonate hydrate, cerium(III) carbonate hydrate, cerium(III) fluoride, cerium(III) chloride, cerium(III) chloride heptahydrate, cerium(III) bromide, cerium(III) iodide, cerium(III) nitrate hexahydrate, cerium(III) oxalate hydrate, cerium(III) sulfate, and cerium(III) sulfate hydrate.

22. A method according to claim 19, characterized in that the heating is performed at 60 to 80°C.

23. A method according to claim 19, characterized in that the cooling is performed by reducing the temperature to 35 to 50°C.

24. A method according to claim 19, characterized in that the inorganic salt is selected from the group consisting of NaCl, CaCl2, KCl, MgCl2, and combinations thereof.

25. A method according to claim 19, characterized in that the inorganic salt solution is a NaCl solution.

26. A method according to claim 25, characterized in that the NaCl solution has a concentration of 0.15 to 0.35 M.

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