Composition for preventing, ameliorating or treating neurodegenerative disease comprising fexofenadine

Abstract

The present invention relates to a composition for preventing, ameliorating or treating neurodegenerative diseases comprising fexofenadine. Fexofenadine is a compound which has been popularized for pharmaceutical use, the safety of fexofenadine has been proved, and when fexofenadine is administered to a neurodegenerative disease model, it is confirmed that the behavioral ability is significantly improved, and the fexofenadine has an effect of protecting nerve cells. The present invention has the effect of inhibiting, for example, the death of dopaminergic neurons, the inflammation-mediated damage of nerve cells or the degradation of nerve cells, the formation of alpha-synuclein aggregates, the abnormal increase in the number of astrocytes or the activation of astrocytes, and the like, and also has the effect of prolonging the life of a neurodegenerative disease model. Therefore, the composition according to the present invention can be effectively used for preventing, ameliorating or treating neurodegenerative diseases.

Description

Technical Field

[0001] The present invention relates to a composition comprising fexofenadine for preventing, improving or treating neurodegenerative diseases. Background Art

[0002] As people age, the incidence of various nervous system and brain-related diseases or neurodegenerative diseases such as dementia, cerebrovascular disease, Parkinson's disease, depression, and Alzheimer's disease is increasing, and as we enter an aging society, their prevalence is expected to increase further. Neurodegenerative diseases are diseases associated with the progressive loss of structure or function of nerve cells (including the death of nerve cells). In particular, it is well known that the onset of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease is closely related to the death of nerve cells.

[0003] In particular, Parkinson's disease is a degenerative disease of the central nervous system caused by hepatic cerebral degeneration or arteriosclerotic changes, and its main symptom is movement disorder, which is caused by the degeneration of nerve cells in a part of the brain called the substantia nigra and the lack of dopamine, a neurotransmitter produced from this area. Parkinson's disease is a chronic, progressive motor nervous system disease. When nerve cells that produce dopamine are damaged, resulting in dopamine deficiency, and acetylcholine that maintains balance becomes dominant, the main symptoms of these diseases are tremors such as shaking of the hands, arms, legs, face, etc., stiffness of the limbs or body, bradykinesia where movements become slow, and unstable postures with lack of balance.

[0004] In addition, Alzheimer's disease is the most common form of dementia in the elderly, showing histopathological characteristics such as overall brain atrophy, ventricular dilatation, neurofibrillary tangles and neuritic plaques, and leading to the gradual decline of intellectual functions such as memory, judgment, language ability, and disorders of daily living ability, personality, and behavior patterns.

[0005] Currently, various treatment methods such as cell transplantation and surgery have been proposed to treat neurodegenerative diseases, but most of them have risk factors and side effects. In addition, due to the complexity of the cell damage mechanism, therapeutic drugs that can substantially inhibit the damage or death of nerve cells have not yet been developed.

[0006] On the other hand, fexofenadine is an antihistamine used to treat allergic symptoms such as seasonal allergies and chronic urticaria (Compalati, E et al. (2011). International Archives of Allergy and Immunology. 156 (1): 1-15.), currently sold under the brand name of Allegra. Therapeutically, fexofenadine acts as a selective peripheral H1 blocker and is classified as a second-generation antihistamine because it does not cross the blood-brain barrier and has less sedative effects than first-generation antihistamines.

[0007] In the present invention, it was confirmed through histological analysis as an animal model of Parkinson's disease (animal model of neurodegenerative disease) that fexofenadine, which has been approved by the FDA as an antihistamine and has been proven to be safe, can inhibit the damage or death of nerve cells, and it was confirmed in an animal model of neurodegenerative disease to which fexofenadine was administered that behavioral ability can actually be improved, thereby demonstrating for the first time the therapeutic effect of fexofenadine on neurodegenerative diseases. Summary of the invention

[0008] Technical issues

[0009] The object of the present invention is to provide a new use of fexofenadine.

[0010] Solutions to the problem

[0011] In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating neurodegenerative diseases, comprising fexofenadine or a pharmaceutically acceptable salt thereof as an effective ingredient.

[0012] In the pharmaceutical composition, it is characterized in that the neurodegenerative disease can be selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, Creutzfeldt-Jakob disease, Lou Gehrig's disease, spinal muscular atrophy, Niemann-Pick disease, synucleinopathy and dementia.

[0013] The present invention is characterized in that the pharmaceutical composition may further comprise a pharmaceutically acceptable excipient.

[0014] The present invention is characterized in that the pharmaceutical composition can protect nerve cells from neurotoxicity.

[0015] The present invention is characterized in that the pharmaceutical composition can protect nerve cells in the following ways:

[0016] (i) inhibiting the death of dopaminergic neurons;

[0017] (ii) inhibiting inflammation-mediated neuronal damage or neuronal degeneration;

[0018] (iii) inhibiting the formation of α-synuclein Ser-129 phosphorylated aggregates; and / or

[0019] (iv) inhibiting the abnormal increase in the number and / or activation of astrocytes.

[0020] The present invention is characterized in that the pharmaceutical composition (i) can improve the exercise capacity of the individual to which the pharmaceutical composition is administered; and / or (ii) can prolong the life span of the individual to which the pharmaceutical composition is administered.

[0021] The present invention also provides a food composition comprising fexofenadine.

[0022] The food composition is characterized in that the food can be a health functional food for preventing or improving neurodegenerative diseases.

[0023] The food composition is characterized in that the neurodegenerative disease can be selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, Creutzfeldt-Jakob disease, Lou Gehrig's disease, spinal muscular atrophy, Niemann-Pick disease, synucleinopathy and dementia.

[0024] The present invention also provides a composition for protecting nerve cells comprising fexofenadine as an effective ingredient.

[0025] Effects of the Invention

[0026] According to the present invention, fexofenadine has been popularized as a compound for pharmaceutical use, and its safety has been proven. When fexofenadine is administered to a neurodegenerative disease model, a significant improvement in behavioral ability is confirmed. It has the effect of protecting nerve cells, such as inhibiting the death of dopaminergic neurons, inhibiting inflammation-mediated nerve cell damage or nerve cell degeneration, inhibiting the formation of α-synuclein aggregates, and / or inhibiting the abnormal increase in the number and / or activation of astrocytes, and has the effect of prolonging the lifespan of a neurodegenerative disease model. Therefore, the composition according to the present invention can be effectively used to prevent, improve or treat neurodegenerative diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The diagram shows the 6-OHDA injection site used to create Parkinson's disease model (6-OHDA model) mice and the fexofenadine administration and analysis schedule.

[0028] Figure 2a It is the result of a bridging test to confirm the therapeutic effect of fexofenadine administration in Parkinson's disease model (6-OHDA model) mice (*p<0.05, **p<0.01).

[0029] Figure 2b The results of the pole test were used to confirm the therapeutic effect of fexofenadine administration in Parkinson's disease model (6-OHDA model) mice (*p<0.05, **p<0.01, ***p<0.001).

[0030] Figure 2c The results of the cylinder test for confirming the therapeutic effect of fexofenadine administration in Parkinson's disease model (6-OHDA model) mice (*p<0.05, **p<0.01, ***p<0.001).

[0031] Figure 3 These are the results of analyzing the ratio of TH-positive neurons and the number of TH-positive neurons in the substantia nigra to confirm the therapeutic effect according to fexofenadine administration in Parkinson's disease model (6-OHDA model) mice (*p<0.05, **p<0.01, ***p<0.001; and Scale bar: 50 μm).

[0032] Figure 4 The results of analyzing the density of TH-positive neurons in the striatum to confirm the therapeutic effect according to fexofenadine administration in Parkinson's disease model (6-OHDA model) mice (*p<0.05, **p<0.01, ***p<0.001; and Scale bar: 50 μm).

[0033] Figure 5 The results of analyzing the number of activated microglia by Iba-1 staining to confirm the therapeutic effect according to fexofenadine administration in Parkinson's disease model (6-OHDA model) mice (*p<0.05, **p<0.01, ***p<0.001; and Scale bar: 50 μm).

[0034] Figure 6 The diagram shows the α-synuclein injection site and the fexofenadine administration and analysis schedule for preparing Parkinson's disease model (α-synuclein injection model) mice.

[0035] Figure 7aThe results of staining of α-synuclein Ser-129 phosphorylated aggregates in the cingulate cortex (Cg), primary motor cortex (M1), secondary motor cortex (M2), caudate putamen (CPu), perirhinal cortex (PRh), midbrain reticular nucleus (MRN), and substantia nigra pars compacta (SNc) on the ipsilateral and contralateral sides of α-synuclein injection were used to confirm the therapeutic effect of fexofenadine administration in Parkinson's disease model (α-synuclein injection model) mice (Scale bar: 50 μm).

[0036] Figure 7b The results are the results of quantitative analysis of the amount of pSer129 a-syn pathology in brain tissues of various sites on the ipsilateral and contralateral sides of the α-synuclein injection to confirm the therapeutic effect of fexofenadine administration in Parkinson's disease model (α-synuclein injection model) mice (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0037] Figure 8 The results of GFAP staining in the cortex, striatum, and substantia nigra on the ipsilateral and contralateral sides of α-synuclein injection were used to confirm the therapeutic effect of fexofenadine administration in Parkinson's disease model (α-synuclein injection model) mice (*p<0.05, **p<0.01, ***p<0.001; and scale bar: 50 μm).

[0038] Fig. 9 The results of analysis of the number of activated microglia by Iba-1 staining in the cortex, striatum, and substantia nigra on the ipsilateral and contralateral sides of α-synuclein injection were used to confirm the therapeutic effect of fexofenadine administration in Parkinson's disease model (α-synuclein injection model) mice (*p<0.05, **p<0.01, ***p<0.001; and scale bar: 50 μm).

[0039] Fig.10a The results are as follows, confirming whether fexofenadine inhibits the aggregate formation of α-synuclein in the BIFC-a-syn C. elegans Parkinson's disease model by Venus fluorescence per unit area (*p<0.05).

[0040] Fig.10b The Venus fluorescence intensity was used to confirm whether fexofenadine inhibited the aggregate formation of α-synuclein in the BIFC-a-syn C.elegans Parkinson's disease model (*p<0.05).

[0041] Fig.10c Pharyngeal pumping analysis was performed to confirm whether fexofenadine restored motor function in the BIFC-a-syn C.elegans Parkinson's disease model (*p<0.05).

[0042] Fig.10d This is the result of life-span analysis to confirm whether fexofenadine treatment has an effect on the life span of the C.elegans Parkinson's disease model in the BIFC-a-syn C.elegans Parkinson's disease model (*p<0.05). DETAILED DESCRIPTION

[0043] Unless otherwise defined, all technical terms and scientific terms used in this specification have the same meanings as those commonly understood by those skilled in the art to which the invention belongs. Generally speaking, the nomenclature used in this specification is well known and commonly used in the art.

[0044] In the present invention, the term "about" or "approximately" refers to within an acceptable error range of a specific value (Value) determined by a person of ordinary skill in the art to which the present invention belongs, and depends in part on the limitations of the measurement system, that is, how to measure or determine the value. For example, according to the implementation of the technical field to which the present invention belongs, "about" may refer to within a standard deviation of 1 or more than 1. Exemplarily, "about" may refer to a range of less than 20%, less than 10%, less than 5%, or less than 1% of a given value or range. Exemplarily, particularly with respect to biological systems or processes, the term may refer to a size within 5 times or within 2 times of a specific value. When a specific value is recorded in the scope of protection of the present application and the invention, unless otherwise stated, it should be assumed that the term "about" means within an acceptable error range of the specific value.

[0045] In the present invention, "administration" of a compound refers to providing the compound or a prodrug of the compound to a subject in need of treatment.

[0046] In the present invention, when fexofenadine is administered to an animal model of neurodegenerative disease, the behavioral ability of the animal model of neurodegenerative disease is restored to a normal level, and it can be confirmed in histological analysis that it has the effect of protecting dopaminergic neurons and inhibiting inflammation-mediated damage to nerve cells and degeneration of nerve cells.

[0047] Therefore, one aspect of the present invention relates to a pharmaceutical composition for preventing or treating a neurodegenerative disease, comprising fexofenadine or a pharmaceutically acceptable salt thereof as an effective ingredient.

[0048] In the present invention, “fexofenadine” may be a compound represented by Formula 1 below.

[0049] Formula 1

[0050]

[0051] In the present invention, "pharmaceutically acceptable salt" refers to a salt that retains the biological effects and properties of a free base or free acid, rather than being biologically unpreferable or otherwise unpreferable. The salt includes: an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, and in particular, hydrochloric acid; and an organic acid, such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and N-acetylcysteine, and the like. In addition, the salt can be prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, and magnesium salts, and the like. Salts derived from organic bases include, but are not limited to, salts of naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, and polyimine resins, and primary, secondary, and tertiary amines, substituted amines.

[0052] The compounds of the present invention may contain multiple asymmetric centers and may exist in the form of optically pure enantiomers, mixtures of enantiomers, such as racemates, optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates or mixtures of diastereomeric racemates. According to the Cahn-Ingold-Prelog rule, an asymmetric carbon atom may be in the "R" or "S" configuration.

[0053] In the present invention, "neurodegenerative disease" refers to a disease associated with symptoms caused by damage to nerve cells, degeneration of nerve cells, loss of function of nerve cells, and / or death of nerve cells, and is characterized in that it can be selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, Creutzfeldt-Jakob disease, Lou Gehrig's disease, spinal muscular atrophy, Niemann-Pick disease, synucleinopathy and dementia, but is not limited to this.

[0054] In the present invention, the dementia may be senile dementia, diabetic dementia, alcoholic dementia or vascular dementia, but is not limited thereto.

[0055] The pharmaceutical composition of the present invention may also include various excipients, including pharmaceutically acceptable diluents or carriers. In some embodiments, the pharmaceutical composition of the present invention may be provided by a method for applying to a subject as needed. In some embodiments, the pharmaceutical composition of the present invention may be applied to humans.

[0056] The pharmaceutical composition of the present invention can be administered to a patient alone or in combination. Coadministration refers to the simultaneous or sequential administration of a compound alone or in combination (of more than one compound or agonist). Therefore, the preparation can also be combined with other active substances when necessary.

[0057] The pharmaceutical composition of the present invention can be prepared according to conventional methods into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories or sterile injections for use.

[0058] The pharmaceutical composition of the present invention can be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally or topically) according to the desired method.

[0059] Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations can be prepared by mixing at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition, in addition to simple excipients, lubricants such as magnesium styrene, talc, etc. can also be used.

[0060] Liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc., which may contain various excipients such as wetting agents, sweeteners, flavors, preservatives, etc. in addition to commonly used simple diluents such as water and liquid paraffin.

[0061] Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As non-aqueous solutions and suspensions, vegetable oils (e.g., propylene glycol, polyethylene glycol, olive oil, etc.), injectable esters (e.g., ethyl oleate, etc.), etc. can be used.

[0062] The pharmaceutical composition can be a sterile injectable preparation, such as a sterile injectable aqueous or oily suspension. The suspension can be prepared using a suitable dispersant or wetting agent (e.g., Tween 80) and a suspending agent according to techniques known in the art. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent (e.g., a solution in 1,3-butanediol). Acceptable carriers and solvents include mannitol, water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils can generally be used as solvents or suspension media. For this purpose, any fixed oil with little irritation can also be used, including synthetic monoglycerides or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives can be used in injectable preparations, as can pharmaceutically acceptable natural oils (e.g., olive oil or castor oil), especially their polyoxyethylated forms.

[0063] The pharmaceutical compositions of the present invention can also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients that are solid at room temperature but liquid at rectal temperature. These substances include, but are not limited to, cocoa butter, beeswax, and polyethylene glycol.

[0064] When the desired treatment is related to a part or organ that is easily accessible by topical application, the parenteral administration of the pharmaceutical composition according to the present invention is particularly useful. When applied topically to the skin, the pharmaceutical composition needs to be formulated into a suitable ointment containing an active ingredient suspended or dissolved in a carrier. The carrier for topical administration of the compound of the present invention includes but is not limited to mineral oil, liquid paraffin, white vaseline, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax and water. In another embodiment, the pharmaceutical composition of the present invention can be formulated into a suitable lotion or cream containing an active compound suspended or dissolved in a carrier. Suitable carriers include but are not limited to mineral oil, sorbitan monostearate, polysorbate 60, myristyl alcohol fatty acid ester (Cetyl esters wax), cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical composition of the present invention can also be topically applied to the lower intestinal tract by rectal suppositories or by suitable enema. Topically applied transdermal patches are also included in the present invention.

[0065] The pharmaceutical compositions of the present invention can be administered by intranasal aerosol or inhalation. These compositions can be prepared according to techniques well known in the art and prepared as saline solutions using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons and / or other solubilizing agents or dispersants known in the art.

[0066] The relative amounts of the active ingredient, pharmaceutically acceptable excipient, and / or any additional ingredients in the pharmaceutical composition of the present invention vary depending on the identity, size, and / or disorder of the subject being treated and on the route of administration of the composition. For example, the content of the active ingredient contained in the pharmaceutical composition of the present invention is not particularly limited thereto, but based on the total weight of the final composition, its content may be 0.0001% to 100% by weight, for example, 0.001% to 50% by weight, and more preferably 0.01% to 10% by weight.

[0067] In the present invention, pharmaceutically acceptable excipients include any and all solvents, dispersion media, diluents, or other liquid carriers, dispersion or suspension aids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc. suitable for the purpose of a specific administration form. Remington's literature [The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro, (Lippincott, Williams & Wilkins, Baltimore, MD, 2006] discloses various excipients for preparing pharmaceutical compositions and known techniques for preparing them. It is considered that any conventional carrier medium is within the scope of the present invention unless it provides any undesirable biological effect or interacts with any other component of the pharmaceutical composition in a harmful manner and cannot coexist with the substance or its derivatives. The purity of the pharmaceutically acceptable excipient is at least 95%, 96%, 97%, 98%, 99%, or 100%.

[0068] The excipient is approved for human and veterinary use. In certain embodiments, the excipient is approved by the U.S. Food and Drug Administration. In certain embodiments, the excipient is pharmaceutical grade. In certain embodiments, the excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and / or the International Pharmacopoeia (EP).

[0069] Pharmaceutically acceptable excipients used to prepare pharmaceutical compositions include, but are not limited to, inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants, and / or oils.

[0070] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium lactose phosphate, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and combinations thereof.

[0071] Exemplary granulating and / or dispersing agents include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (propolis), sodium lauryl sulfate, quaternary ammonium compounds, and combinations thereof.

[0072] Exemplary surfactants and / or emulsifiers include, but are not limited to, natural emulsifiers (e.g., gum arabic, agar, alginic acid, sodium alginate, tragacanth, Chondrux, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, lanolin, cholesterol, waxes, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and propolis [magnesium aluminum silicate]), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), glue, cellulose derivatives (e.g. sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan fatty acid esters (e.g. polyoxyethylene sorbitan monolaurate [Tween 20], polyoxyethylene sorbitan [Tween 60], polyoxyethylene sorbitan monooleate [Tween 80], sorbitan monopalmitate [Span 40], sorbitan monostearate [Span 60], sorbitan tristearate [Span 65], glycerol monooleate, sorbitan monooleate [Span 80]), polyoxyethylene esters (e.g. polyoxyethylene monostearate [Myr 45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate and polyethylene glycol stearate (Solutol)), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., polyoxyethylene hydrogenated castor oil (Cremophor)), polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether [brij 30]), poly(vinyl pyrrolidone), diethylene glycol laurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, lauryl ethyl ester, sodium lauryl sulfate, Pluronic F68, poloxamer 188, cetrimide bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and / or combinations thereof.

[0073] Exemplary binders include: starches (e.g., corn starch and starch paste); gelatin; sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol); natural and synthetic gums (e.g., acacia, sodium alginate, Irish moss extract, panwar gum, ghatti gum, mucilage of isapol husk, carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl pyrrolidone), magnesium aluminum silicate (propolis), and Larch Arabinogalactan); alginates; polyethylene oxide; polyethylene glycol; inorganic calcium salts; silicic acid; polymethacrylates; waxes; water; alcohols; and combinations thereof, but are not limited thereto.

[0074] Exemplary preservatives may also include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives and other preservatives. Exemplary antioxidants include, but are not limited to, alpha-tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium pyrosulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium pyrosulfite and sodium sulfite. Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid and trisodium edetate. Exemplary antimicrobial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethanol, glycerol, hexetidine, imidurea, phenol, phenoxyethanol, phenylethanol, phenylmercuric nitrate, propylene glycol, and thimerosal. Exemplary antifungal preservatives include, but are not limited to, butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid. Exemplary alcohol preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoates, and phenylethanol. Exemplary acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid. Other preservatives include, but are not limited to, tocopherol, tocopherol acetate, deteroxime mesylate, cetrimonium bromide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and butylated hydroxytoluene. (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus, Phenonip, methyl paraben, imidazolidinyl urea (GERMALL 115), GERMABEN II, NEOLONE, Kathon and Euxyl. In a specific embodiment, the preservative is an antioxidant. In another embodiment, the preservative is a chelating agent.

[0075] Exemplary buffers include, but are not limited to, citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glucuronate, calcium glucoheptonate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propionic acid, calcium levulinate, valeric acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, dibasic calcium phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol, and combinations thereof.

[0076] Exemplary lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silicon dioxide, talc, malt, glyceryl behenate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and combinations thereof.

[0077] Exemplary oils include, but are not limited to, almond oil, apricot kernel oil, avocado oil, babassu oil, bergamot oil, black currant seed oil, borage oil, juniper berry oil, chamomile oil, canola oil, coriander oil, Brazilian palm oil, castor oil, cinnamon oil, cocoa butter, coconut oil, cod liver oil, coffee oil, corn oil, cottonseed oil, emu oil, eucalyptus oil, evening primrose oil, fish oil, linseed oil, geraniol, gourd oil, grape seed oil, hazelnut oil, hyssop oil, isopropyl myristate, jojoba oil, macadamia oil, lavender oil (l

[0013] The oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and combinations thereof.

[0078] These excipients may also be optionally included in the formulations of the present invention. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may be present in the composition according to the judgment of the formulator.

[0079] The term "preventing" of the present invention includes preventing or delaying the onset of future clinical or subclinical symptoms of a disease, disorder or condition in a mammal, particularly, in an individual who may be at risk of or susceptible to the disease, disorder or condition but who has not yet experienced or displayed clinical or subclinical symptoms of the disease, disorder or condition.

[0080] The term "treatment" of the present invention includes:

[0081] (1) inhibiting a condition, disorder or condition (e.g., inhibiting, reducing or delaying the recurrence of at least one clinical or subclinical symptom during onset or maintenance treatment of a disease); and / or

[0082] (2) Alleviating a disease, disorder or condition (i.e., causing regression of the disease, disorder or condition, or at least one of its clinical or subclinical symptoms).

[0083] The benefit conferred to the treated patient may be statistically significant or at least perceptible to the patient or physician. However, those skilled in the art will appreciate that when a drug is administered to a patient to treat a disease, the result may not always be an effective treatment.

[0084] Although the pharmaceutical compositions provided in the present invention are in principle directed to pharmaceutical compositions for administration to humans, it will be appreciated by those skilled in the art that these compositions are generally suitable for administration to animals of all types. That is, the pharmaceutical compositions according to the present invention may also be administered to animals in need of veterinary treatment, such as livestock (e.g., dogs, cats, etc.), farm animals (e.g., cattle, sheep, pigs, horses, etc.), and other mammals such as laboratory animals (e.g., rats, mice, guinea pigs, etc.). The modifications of the pharmaceutical compositions administered to various animals are well understood and skilled veterinary pharmacologists can design and / or these modifications by simple experiments when necessary.

[0085] The pharmaceutical compositions described in the present invention can be prepared by methods known in the art of pharmacology or discussed in any subsequent content. Generally, these tableting methods include the steps of associating the active ingredient with excipients and / or one or more other auxiliary ingredients, followed by the steps of forming and / or packaging the product into the desired single or multiple dose units when necessary or desired.

[0086] The pharmaceutical composition of the present invention can also be prepared, packaged and / or sold in an unpackaged state as a single unit dose and / or multiple single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition containing a preset amount of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient administered to the subject and / or a convenient division of the dose, such as 1 / 2 or 1 / 3 of the dose.

[0087] In the present invention, "administration" means introducing a predetermined substance into a patient's body by any suitable method, and the administration route of the pharmaceutical composition can be administered by any general route as long as the drug can reach the target tissue. The administration route may include, but is not limited to, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, rectal administration, etc.

[0088] The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

[0089] The term "pharmaceutically effective amount" of the present invention may refer to a "therapeutically-effective amount", which refers to an amount of a compound or composition (e.g., a compound or composition of the present invention) sufficient to achieve a beneficial or desired result. A pharmaceutically effective amount may be more than one administration or application or administration in a dosage form and is not intended to be limited to a specific formulation or route of administration.

[0090] The effective dosage level can be determined based on the severity of the disease, the activity of the drug, the patient's age, weight, health condition, sex, the patient's sensitivity to the drug, the administration time, administration route and excretion rate of the composition of the present invention used during treatment, factors including drugs used in combination or concomitantly with the composition of the present invention used, and other factors well known in the medical field.

[0091] The pharmaceutical composition of the present invention can be administered as a single therapeutic drug or in combination with other therapeutic drugs, and can be administered sequentially or simultaneously with conventional therapeutic drugs. And it can be administered once or multiple times. It is important to consider all the above factors when administering, and its dosage can achieve the maximum effect with the minimum amount without side effects.

[0092] A person skilled in the art can determine the dosage of the pharmaceutical composition of the present invention by considering the purpose of use, the degree of addiction of the disease, the age, weight, sex, past medical history of the patient, or the type of substance used as the active ingredient, etc. For reference, Allegra is orally administered in tablet form, 120 mg each time, once a day, with water.

[0093] The present invention is characterized in that the pharmaceutical composition of the present invention containing fexofenadine as an active ingredient can protect nerve cells from neurotoxicity.

[0094] For example, it is characterized in that the pharmaceutical composition can protect nerve cells by: (i) inhibiting the death of dopaminergic neurons; (ii) inhibiting inflammation-mediated nerve cell damage or nerve cell degeneration; (iii) inhibiting the formation of α-synuclein Ser-129 phosphorylated aggregates; and / or (iv) inhibiting the abnormal increase in the number and / or activation of astrocytes.

[0095] For example, the present invention is characterized in that fexofenadine can inhibit the activation of microglia to have the effect of inhibiting inflammation-mediated nerve cell damage and nerve cell degeneration and protecting nerve cells.

[0096] Furthermore, the present invention is characterized in that the pharmaceutical composition comprising fexofenadine as an active ingredient (i) improves the motor ability of an individual to whom the pharmaceutical composition is administered; and / or (ii) prolongs the life span of an individual to whom the pharmaceutical composition is administered, but is not limited thereto.

[0097] Another aspect of the present invention relates to a method for preventing or treating a neurodegenerative disease, the method comprising the step of administering fexofenadine to an individual in need thereof.

[0098] Another aspect of the present invention relates to the use of fexofenadine for preventing or treating neurodegenerative diseases.

[0099] Another aspect of the present invention relates to the use of fexofenadine in preparing a medicament for preventing or treating a neurodegenerative disease.

[0100] The term "subject" of the present invention refers to all animals, including humans, that have developed or are likely to develop a neurodegenerative disease, and by administering the composition of the present invention to the subject, the disease can be effectively prevented and / or treated.

[0101] Still another aspect of the present invention relates to a food composition comprising fexofenadine.

[0102] In a preferred embodiment, the food may be a health functional food for preventing or improving neurodegenerative diseases.

[0103] The term "improvement" according to the present invention refers to all actions which at least reduce a parameter associated with the state being treated, for example all actions which at least reduce the extent of a symptom.

[0104] In the food composition of the present invention, "neurodegenerative disease" refers to a disease associated with the symptoms manifested by damage to neurons, degeneration of neurons, loss of function of neurons, and / or death of neurons, characterized in that it is selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, Creutzfeldt-Jakob disease, Lou Gehrig's disease, spinal muscular atrophy, Niemann-Pick disease, synucleinopathy and dementia, but is not limited thereto.

[0105] In the food composition of the present invention, the dementia may be senile dementia, diabetic dementia, alcoholic dementia or vascular dementia, but is not limited thereto.

[0106] When using the food composition of the present invention as a food additive, the composition can be directly added, or used together with other foods or food ingredients, and can be appropriately used according to a conventional method. Usually, when preparing food or beverage, relative to raw materials, the composition of the present invention can be added in an amount of less than 15 % by weight, preferably, it can be added in an amount of less than 10 % by weight. However, when long-term intake for the purpose of health or for the purpose of health control, it can be added in an amount of less than 5 % by weight, because there is no problem in terms of safety, therefore it is also possible to add active ingredients in an amount above the above range.

[0107] The food composition of the present invention can be prepared in all forms such as food additives, nutritional supplements, and health functional foods.

[0108] For example, as health functional food, the composition of the present invention can be prepared in the form of tea, fruit juice and beverage to drink, or can be taken in by granulation, encapsulation and powderization. In addition, the composition of the present invention can be added to health beverages (including alcoholic beverages), fruits and processed foods thereof (such as canned fruit, bottled canned food, jam (Jam), marmalade (Marmalade) etc.), fish, meat and processed foods thereof (such as ham, sausage, corned beef etc.), breads and noodles (such as udon noodles, buckwheat noodles, ramen noodles, pasta, macaroni etc.), fruit juice, various beverages, biscuits, toffee, dairy products (such as butter, cheese etc.), edible vegetable oils, margarine, vegetable protein, steamed food, frozen food and various seasonings (such as soybean paste, soy sauce, sauce etc.) etc. to prepare.

[0109] When fexofenadine is added to a health drink for preparation, it may contain various flavoring agents or natural carbohydrates as additional ingredients, just like ordinary drinks. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, or synthetic sweeteners such as saccharin and aspartame. The ratio of the natural carbohydrates may be appropriately determined by the choice of a person skilled in the art to which the present invention belongs.

[0110] In addition to the above, food composition of the present invention can contain various nutrients, vitamins, electrolytes, spices, coloring agents, pectin acid and its salt, alginic acid and its salt, organic acid, protective colloid thickener, pH adjusting agent, stabilizer, preservative, glycerine, alcohol, carbonating agent for carbonated beverage etc. In addition, food composition of the present invention can contain the pulp for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients can be used alone or in combination. The ratio of these additives can also be suitably selected by those of ordinary skill in the technical field to which the present invention belongs.

[0111] Another aspect of the present invention relates to a composition for protecting nerve cells comprising fexofenadine as an effective ingredient.

[0112] In the compositions according to various embodiments of the present invention, the preventive, therapeutic and improving effects of fexofenadine can also be applied to neurodegenerative diseases caused by various reasons.

[0113] In the present invention, "individual", "patient" and "subject" are used interchangeably and include any animal, including mammals, such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates including humans.

[0114] Example

[0115] The present invention is further described in detail below by way of examples. These examples are only used to exemplify the present invention, and it is obvious to those skilled in the art that the scope of the present invention should not be construed as being limited by these examples.

[0116] Example 1. Preparation of Parkinson's disease animal model (6-OHDA model) and drug administration

[0117] Hydroxydopamine, also known as 6-hydroxydopamine (6-OHDA) or 2,4,5-trihydroxyphenylethylamine, is a neurotoxic synthetic organic compound that researchers use to selectively destroy dopaminergic and noradrenergic neurons in the brain and to develop and test new drugs and treatments for Parkinson's disease. It is known that Parkinson's disease is induced in experimental animals by destroying dopaminergic neurons in the substantia nigra (see Simola, Nicola et al. (2007). "The 6-Hydroxydopamine model of parkinson's disease". Neurotoxicity Research. 11(3): 151-167.). Accordingly, an animal model of Parkinson's disease was created by injecting 6-OHDA.

[0118] Specifically, 8-week-old C57BL / 6 mice (DaeHan Biolink, Chungcheongbuk-do, Eumseong, Korea) were anesthetized by intraperitoneal (ip) injection of 2,2,2-tribromoethanol (Sigma-Aldrich) 200 mg / kg and placed in a stereotaxic instrument (David Kopf Instruments, Tujunga, CA, USA) equipped with a mouse anesthesia adapter. A 6-OHDA-HCl solution in 0.02% ascorbic acid was added using a Hamilton syringe (Hamilton, Bonaduz, Switzerland). (Sigma Aldrich) Injections were made into the right striatum at the following stereotaxic coordinates (mm from the anterior vertex): AP +1.0; ML -1.8; DV -3.2. The skin was sutured and the animals were removed from the stereotaxic apparatus and placed on a heating pad for 30 minutes. Normal control mice of the disease mouse model were injected with vehicle (0.02% ascorbic acid) instead of 6-OHDA-HCl solution.

[0119] Next, after a 3-day recovery period after surgery, the disease model or normal control mice were orally administered phosphate buffered saline (PBS) (drug control group) or 20 mg / kg of fexofenadine (Sigma Aldrich) dissolved in PBS once a day and the experiment was performed. Behavioral experiments were performed 18 days after the injection of 6-OHDA, and histological analysis was performed after the mice were killed 21 days after the injection of 6-OHDA.

[0120] The experiment was divided into the following 4 groups:

[0121] Normal mouse control group (PBS-Control): after injection of vehicle (Vehicle), PBS was administered (n=13);

[0122] Normal mouse drug administration group (PBS-fexofenadine): Fexofenadine was administered after injection of vehicle (Vehicle) (n=5);

[0123] Disease mouse control group (6OHDA): PBS administration after injection of 6-OHDA (n=12); and

[0124] Disease mouse drug administration group (6OHDA+fexofenadine): after the injection of 6-OHDA, fexofenadine was administered (n=13).

[0125] Example 2. Behavior test in the Parkinson's disease animal model (6-OHDA model)

[0126] 2-1. Bridging test

[0127] In order to test the balance sense of the animal model, a bridging test was performed to measure the time required to cross from one side to the other side of a 1 m long pole (crossing time).

[0128] The results, such as Figure 2a As shown, the crossing time of the disease mouse control group (6OHDA) increased by about 2 times compared with the normal mouse control group (PBS-Control), but the crossing time in the disease mouse drug administration group (6OHDA+fexofenadine) in which fexofenadine was administered to the Parkinson's disease animal model was restored to a level similar to that of the normal mouse control group (PBS-Control).

[0129] On the other hand, it was confirmed that the crossing time of the normal mouse drug administration group (PBS-fexofenadine) administered with fexofenadine to normal mice was improved compared with the normal mouse control group (PBS-Control), indicating that fexofenadine also has the effect of improving the sense of balance in normal individuals.

[0130] 2-2. Pole climbing test

[0131] The mice were placed on a pole with a diameter of 8 mm and a length of 55 cm, and a pole climbing test was performed to measure the time required for the mice to land on all fours (landing time).

[0132] The results, such as Figure 2bAs shown, the landing time of mice in the disease mouse control group (6OHDA) was about 8 to 9 seconds, which was significantly reduced compared with the landing time of about 6 seconds in the normal mouse control group (PBS-Control). The landing time of the disease mouse drug administration group (6OHDA+fexofenadine) was about 6 seconds. It can be confirmed that the administration of fexofenadine can restore the landing ability of Parkinson's disease model mice to normal levels.

[0133] On the other hand, the landing time of the normal mouse drug administration group (PBS-fexofenadine) was about 4.5 seconds, confirming that the landing ability was improved compared with the normal mouse control group (PBS-Control), thus reconfirming that fexofenadine also has the effect of improving behavioral ability in normal individuals.

[0134] 2-3. Cylinder experiment

[0135] The cylinder test was performed by placing the mouse in a beaker with a diameter of 20 cm and a height of 40 cm, and the number of times the mouse stood up and stepped on the beaker wall with its front paws was recorded. The results were expressed as a percentage (%) of the number of steps made by the front paw on the opposite side of the damaged striatum compared to the front paw on the opposite side of the undamaged striatum.

[0136] The results, such as Figure 2c As shown, compared with the normal mouse control group (PBS-Control), the number of steppings by the contralateral forefoot of the damaged striatum in the disease mouse control group (6OHDA) was reduced by nearly 10% compared with the contralateral forefoot of the undamaged striatum, but it was confirmed that in the disease mouse drug administration group (6OHDA+fexofenadine), the motor ability recovered to the level of the normal mouse control group (PBS-Control).

[0137] Example 3. Histological analysis in the animal model of Parkinson's disease (6-OHDA model)

[0138] 3-1. Tyrosine Hydroxylase in the Substantia Nigra and Striatum (tyrosinehydroxylase, TH) staining results

[0139] Tyrosine hydroxylase (or tyrosine 3-monooxygenase) is an enzyme that catalyzes the conversion of the amino acid L-tyrosine into L-3,4-dihydroxyphenylalanine (L-DOPA) and is known as a marker for dopamine, norepinephrine, and adrenaline (catecholamine)-containing neural cells and endocrine cells. In order to confirm whether the administration of fexofenadine can protect dopaminergic neurons in an animal model of Parkinson's disease, an experiment to stain TH-positive cells was conducted.

[0140] Specifically, the mouse of Example 1 was anesthetized with urethane, and then the chest was cut open, and the perfusion solution was passed through the left ventricle of the heart for 2 minutes to perform perfusion, thereby isolating the mouse brain. The mouse brain was fixed by immersion in 4% paraformaldehyde, transferred to 30% sucrose, and after dehydration, it was cut into coronal sections with a thickness of 35 μm on a freezing microtome, and was immersed in an antifreeze (25% ethylene glycol, 25% glycerol, and 0.05M phosphate buffer) for storage. Six slices were used for each mouse, including the part of the striatum (from 0.98 mm to -0.10 mm from the apex) and the part of the substantia nigra (from -2.80 mm to -3.88 mm from the apex) for tissue staining.

[0141] Before immunostaining, brain tissue sections were washed three times with PBS in 24 wells and then treated with 3% hydrogen peroxide for 5 minutes to remove endogenous peroxidase activity. Then, for dopaminergic neuron staining, rabbit anti-TH antibody (Abcam, ab137721, 1:1000 dilution) was added to PBS containing 0.2% Triton X-100 and 1% BSA, and then incubated at 4°C overnight. Next, after incubation with secondary antibody (biotinylated anti-rabbit IgG, 1:1000 dilution) at 4°C for 1 hour, ABC solution was added at room temperature and further incubated for 1 hour. Tissue sections were developed with DAB for 2 minutes and then attached to gelatin-coated slides, dehydrated by increasing the ethanol concentration from 70% ethanol to 80%, 90%, and 100%, reacted with xylene for one day, and then mounted.

[0142] Images were taken using an optical microscope (Olympus Microscope System BX51; Tokyo, Japan) and quantified using Image J software (Bethesda, MD, USA). Optical density measurements or cell number measurements were used as the average of the values ​​of 6 sections of tissue for each mouse, expressed as the ratio of the density or number of TH-positive neurons on the right ipsilateral side of the 6-OHDA injury to the TH-positive neurons on the contralateral side of the 6-OHDA injury.

[0143] The results, such as Figure 3As shown, in the disease mouse control group (6OHDA), the ratio of TH-positive neurons in the ipsilateral and contralateral sides of the substantia nigra decreased by about 50%, but in the disease mouse drug administration group (6OHDA + fexofenadine), it was confirmed that the ratio of TH-positive neurons in the ipsilateral and contralateral sides of the substantia nigra recovered to nearly 80%.

[0144] In addition, compared with the normal mouse control group (PBS-Control), the number of TH-positive neurons in the ipsilateral side of the substantia nigra in the disease mouse control group (6OHDA) was significantly reduced, whereas in the disease mouse drug administration group (6OHDA+fexofenadine), it was confirmed that the number of TH-positive neurons was restored to a level similar to that of the normal mouse control group (PBS-Control).

[0145] On the other hand, Figure 4 As shown, a similar pattern was confirmed in the striatum. In the disease mouse control group (6OHDA), the density of TH-positive neurons on the ipsilateral side compared to the contralateral side was reduced by about 50% compared to the normal mouse control group (PBS-Control). Conversely, in the disease mouse drug administration group (6OHDA+fexofenadine), the density of TH-positive neurons on the ipsilateral side compared to the contralateral side was restored to about 80% compared to the normal mouse control group (PBS-Control).

[0146] The above results show that fexofenadine has the effect of inhibiting the death of dopaminergic neurons or protecting dopaminergic neurons by being toxic to nerve cells.

[0147] 3-2. Microglia staining results in the substantia nigra and striatum

[0148] It is known that microglia exhibit a surveillance mode under normal conditions, but as they become activated, they undergo an inflammatory phase and an anti-inflammatory phase in sequence, and induce inflammation-mediated neuronal damage and neuronal degeneration. Therefore, the effect of fexofenadine on microglia activation was analyzed.

[0149] Activated microglia were analyzed by staining with anti-Iba-1 antibody (Wako, 019-19741, 1:1000 dilution). The brain tissue preparation and tissue staining method for this purpose were performed in the same steps as the TH staining method of Example 3-1, and the images were analyzed using an Axio scan.Z1 slide scanner (Carl Zeiss, Germany). The number of cells was measured with the average of 3 images per tissue for analysis.

[0150] The results, such as Figure 5As shown, it was confirmed that the number of Iba-1 positive microglia in the ipsilateral side of the striatum and substantia nigra in the disease mouse control group (6OHDA) increased compared with the normal mouse control group (PBS-Control), whereas the number of Iba-1 positive microglia in the disease mouse drug administration group (6OHDA+fexofenadine) decreased to a level similar to that of the normal mouse control group (PBS-Control).

[0151] The above results confirmed that fexofenadine has the effect of inhibiting the activation of microglia and suppressing the damage and degeneration of neurons mediated by inflammation to protect neurons by inhibiting the activation of microglia.

[0152] Example 4. Preparation of Parkinson's disease animal model (α-synuclein injection model) and drug administration

[0153] It is known that when α-synuclein prepared from recombinant protein is injected into the striatum of mice, endogenous α-synuclein of mice is collected to induce Parkinson's disease (Luk et al. 2012, Science. 2012 Nov 16; 338(6109): 949-53). Therefore, referring to the method described in the literature, α-synuclein was injected to prepare an animal model of Parkinson's disease.

[0154] Specifically, 8-week-old C57BL / 6 mice (DaeHan Biolink, Eumseong, Chungcheongbuk-do, Korea) were anesthetized by intraperitoneal (ip) injection of 2,2,2-tribromoethanol (Sigma-Aldrich) 200 mg / kg and placed in a stereotaxic instrument (David Kopf Instruments, Tujunga, CA, USA) equipped with a mouse anesthesia adapter. α-Synuclein prepared from recombinant protein purified from BL21 Escherichia coli strain transformed with 5 mg / ml of pRK172 α-synuclein plasmid was injected into the mouse anesthesia adapter using a Hamilton syringe (Hamilton, Bonaduz, Switzerland). Injections were made into the right striatum at the following stereotaxic coordinates (mm from the anterior vertex): AP +1.0; ML -1.8; DV -3.2. The skin was sutured and the animals were removed from the stereotaxic apparatus and placed on a heating pad for 30 minutes.

[0155] Then, after a 3-day recovery period after surgery, the disease model mice were orally administered PBS (drug control group) or 20 mg / kg of fexofenadine (Sigma Aldrich) dissolved in PBS once a day and the experiment was carried out. The mice were killed on the 30th day and histological analysis was performed. The control group mice were injected with vehicle (PBS) instead of α-synuclein.

[0156] The experiment was divided into the following three groups:

[0157] Normal mouse control group (PBS): PBS administration after injection of vehicle (Vehicle) (n=3);

[0158] Disease mouse control group (PFF-PBS): PBS was administered after injection of α-synuclein (n=6); and

[0159] Disease mouse drug administration group (PFF-fexofenadine): after injection of α-synuclein, fexofenadine was administered (n=7).

[0160] Example 5. Histological analysis in the Parkinson's disease animal model (α-synuclein injection model)

[0161] 5-1. Staining results of α-synuclein Ser-129 phosphorylated aggregates

[0162] It is known that when α-synuclein is injected into the striatum of mice, the endogenous α-synuclein of the mice is collected and α-synuclein Ser-129 phosphorylated aggregates are formed inside the neurons of the cortex, amygdala and substantia nigra, thereby leading to the death of neurons and damage to nerve function.

[0163] Therefore, an attempt was made to use the Parkinson's disease animal model prepared in Example 4 to confirm whether fexofenadine can inhibit the formation of α-synuclein Ser-129 phosphorylated aggregates.

[0164] To this end, tissue staining was performed using pSer129 a-syn antibody (Wako, 1:5000 dilution) in the cingulate cortex (Cg), primary motor cortex (M1), secondary motor cortex (M2), caudate putamen (CPu), perirhinal cortex (PRh), midbrain reticular nucleus (MRN), and substantia nigra pars compacta (SNc) on the ipsilateral and contralateral sides of the α-synuclein injection using the same method as described in Example 3-2.

[0165] In addition, brain tissues at various locations were isolated on the ipsilateral and contralateral sides of the α-synuclein injection to quantify the amount of pSer129a-syn pathology using Zenblue 3.1 software.

[0166] The results, such as Figure 7a and Figure 7b As shown, in the disease mouse control group (PFF-PBS), it was confirmed that the levels of α-synuclein Ser-129 phosphorylated aggregates were very high in various brain tissues on the ipsilateral and contralateral sides of the α-synuclein injection. However, in the disease mouse drug administration group (PFF-fexofenadine), it was confirmed that the formation of α-synuclein Ser-129 phosphorylated aggregates was inhibited, and this pattern was observed more clearly on the ipsilateral side of the α-synuclein injection.

[0167] From the above results, it can be seen that fexofenadine inhibits the formation of α-synuclein Ser-129 phosphorylated aggregates, thereby inhibiting the death of nerve cells and the damage of nerve function.

[0168] 5-2. Glial fibers in the cortex, striatum, and substantia nigra GFAP staining results

[0169] Glial fibrillary acidic protein (GFAP) is an intermediate microfilament protein of astrocytes. The expression of GFAP is regulated by post-traumatic signals and neural activities at various stages. In particular, GFAP is used as a marker to show the abnormal increase or activation of astrocytes (Astrogliosis) caused by the destruction of nerve cells due to damage or stress of the central nervous system (CNS).

[0170] Therefore, an attempt was made to use the Parkinson's disease animal model prepared in Example 4 to confirm whether fexofenadine can inhibit the abnormal increase in the number or activation of astrocytes.

[0171] For this purpose, each mouse was stained with GFAP antibody (Neuromics, Edina, Minnesota, USA) in the same manner as in Example 3-2, using 6 images each in the area containing the cortex (from 0.98 mm to -0.10 mm from the vertex), the area containing the striatum (from 0.98 mm to -0.10 mm from the vertex), and the area containing the substantia nigra (from -2.8 mm to -3.88 mm from the vertex). Intensity was measured using ImageJ software to measure the difference between the disease mouse control group and the normal mouse control group or between the disease mouse drug administration group and the disease mouse control group.

[0172] The results, such as Figure 8 As shown, it was confirmed that in the disease mouse control group (PFF-PBS), the expression of GFAP in the cortex, striatum, and substantia nigra on the ipsilateral and contralateral sides of the α-synuclein injection was significantly increased compared with the normal mouse control group (PBS), but in the disease mouse drug administration group (PFF-fexofenadine), the expression of GFAP in the cortex, striatum, and substantia nigra on the ipsilateral and contralateral sides of the α-synuclein injection was significantly decreased compared with the disease mouse control group (PFF-PBS).

[0173] From the above results, it was confirmed that fexofenadine has the effect of suppressing the abnormal increase in the number and activation of astrocytes caused by the destruction of nerve cells and protecting nerve cells.

[0174] 5-3. Microglia in the cortex, striatum, and substantia nigra Cell staining results

[0175] An attempt was made to use the Parkinson's disease animal model prepared in Example 4 to analyze the effect of fexofenadine on microglial activation.

[0176] To this end, 6 images were used for each mouse, including the part containing the cortex (from 0.98 mm to -0.10 mm from the apex), the part containing the striatum (from 0.98 mm to -0.10 mm from the apex), and the part containing the substantia nigra (from -2.8 mm to -3.88 mm from the apex), and IBA-1 staining was performed in the same way as in Example 3-2.

[0177] The results, such as Fig. 9 As shown, it was confirmed that in the disease mouse control group (PFF-PBS), the density of IBA-1 positive cells in the cortex, striatum, and substantia nigra on the ipsilateral side to which α-synuclein was injected was significantly increased compared with the normal mouse control group (PBS), but in the disease mouse drug administration group (PFF-fexofenadine), the density of IBA-1 positive cells in the cortex, striatum, and substantia nigra on the ipsilateral side to which α-synuclein was injected was significantly decreased compared with the disease mouse control group (PFF-PBS).

[0178] On the other hand, it was confirmed that in the disease mouse control group (PFF-PBS), the density of IBA-1 positive cells in the cortex and striatum on the contralateral side to which α-synuclein was injected was also significantly increased compared with the normal mouse control group (PBS), but in the disease mouse drug administration group (PFF-fexofenadine), the density of IBA-1 positive cells was reduced compared with the disease mouse control group (PFF-PBS).

[0179] The above results also demonstrate that fexofenadine has the effect of inhibiting the activation of microglia and suppressing the damage and degeneration of neural cells mediated by inflammation, thereby protecting neural cells.

[0180] Example 6. Validation of the Parkinson's disease model in C. elegans using BIFC-a-syn

[0181] It is known that protein aggregates called Lewy bodies are formed in nerve cells of the substantia nigra in the midbrain of Parkinson's disease patients, and α-synuclein aggregates in the Lewy bodies to induce neurotoxicity.

[0182] On the other hand, bimolecular fluorescence complementation (BiFC) analysis is a method based on the reconstruction of two non-fluorescent fragments derived from fluorescent proteins, which are divided into N-terminal Venus (VN) fragment and C-terminal Venus (VC) fragment. When A and B connected to VN and VC are combined, they are reconstructed into a single fluorescent protein Venus, thereby expressing fluorescence. In this case, when α-synuclein is expressed in a manner that is connected to VN and VC, respectively, fluorescence is expressed as α-synuclein forms aggregates.

[0183] Therefore, in the present invention, an attempt was made to use the BiFC-a-syn C.elegans Parkinson's disease model to verify whether fexofenadine inhibits the formation of α-synuclein aggregates.

[0184] To this end, plasmids for overexpression of α-synuclein connected to VN in muscle cells of the pharynx and α-synuclein connected to VC in nerve cells that form synapses with muscle cells were introduced into wild-type BristolN2 C. elegans purchased from the Caenorhabditis Genetics Center (CGC, University of Minnesota, St. Paul, MN) to create a C. elegans Parkinson's disease model (Venus-αS) expressing BiFC-α-syn (Kim D-K et al., (2016). Autophagy. 12(10): 1849-1863).

[0185] The L4 stage C.elegans (Venus-αS) model expressing BiFC-α-syn was transferred to a plate containing DMSO (dimethyl sulfoxide) or 50 μM fexofenadine and cultured in a 20°C incubator for 2 days. The control group and the fexofenadine-treated C.elegans group were added to M9 buffer (22 mM KH2PO4, 22 mM Na2HPO4, 85 mM NaCl, 1 mM MgSO4) containing 10 mM sodium azide for fixation, and after being dispensed into 96-well plates, the intensity of VENUS fluorescence was measured using an intracellular fluorescence scanning device (Cytiva).

[0186] The results, such as Fig.10a and Fig.10bAs shown, the Venus fluorescence intensity in the C. elegans group treated with fexofenadine was significantly reduced compared with the Venus fluorescence intensity in the control group not treated with fexofenadine, and it was confirmed that fexofenadine inhibited the formation of transitional aggregates of α-synuclein.

[0187] On the other hand, pharyngeal pumping analysis was performed to confirm whether treatment with fexofenadine can restore impaired motor ability in the C.elegans Parkinson's disease model. After 8 days of treatment with fexofenadine, pharyngeal pumping was counted using an optical microscope at room temperature for 1 minute, and the data were expressed in PPM (pumps per minute).

[0188] The results, such as Fig.10c As shown, it was confirmed that in the C. elegans Parkinson's disease model, the pharyngeal pumping ability of C. elegans was drastically reduced compared to the wild type N2, but when fexofenadine was treated in the C. elegans Parkinson's disease model, the pharyngeal pumping ability of C. elegans was restored to a level similar to that of the wild type N2. Interestingly, when fexofenadine was treated in the C. elegans wild type N2, it was also confirmed that the pharyngeal pumping ability of C. elegans was improved.

[0189] Finally, in order to confirm the effect of fexofenadine treatment on the life span of the C.elegans Parkinson's disease model, a life span analysis was performed. To this end, eggs laid by adults were grown to the L4 larval stage at 20°C on NGM plates inoculated with E. coli OP50 (CGC). When the L4 stage was reached, it was transferred to an NGM plate containing 100mM 5-fluoro-2'-deoxyuridine (Sigma-Aldrich, F0503) to prevent the production of offspring. The number of C.elegans that survived or died was recorded every 1 or 2 days. Worms that broke or buried or crawled out of the plate were also counted. Survival data were analyzed using OASIS2 (online application for survival analysis for life span analysis http: / / sbi.postech.ac.kr / oasis / surv / ).

[0190] The results, such as Fig.10dAs shown, the life span of C.elegans wild type N2 is about 15 days, the life span of C.elegans Parkinson's disease model is shortened to about 12 days, but the life span of C.elegans Parkinson's disease model treated with fexofenadine is about 14 days, confirming that fexofenadine prolongs the life span of C.elegans Parkinson's disease model. Interestingly, when fexofenadine is treated in C.elegans wild type N2, it is also confirmed that the life span of C.elegans is further extended.

[0191] The specific parts of the content of the present invention are described in detail above. It is clear to those skilled in the art that these specific technologies are only preferred embodiments and do not limit the scope of the present invention. Therefore, the actual scope of the present invention will be defined by the attached invention claims and their equivalents.

[0192] [National R&D Program to Support the Invention]

[0193] [Project unique number] 1711161142

[0194] [Project number] 2019R1A5A2026045

[0195] [Department Name] Ministry of Science and ICT

[0196] [Project Management (Professional) Organization Name] Korea Research Foundation

[0197] [Research Project Name] Leading Research Center Support Project Basic Medical Field (MRC)

[0198] [Research Topic Name] Brain Disease Convergence Research Center

[0199] [Contribution rate] 1 / 1

[0200] [Project implementing organization name] Ajou University, South Korea

[0201] [Study period] 2019.06.01~2026.02.28

Claims

1. A pharmaceutical composition for preventing or treating a neurodegenerative disease, characterized in that: The pharmaceutical composition comprises fexofenadine or a pharmaceutically acceptable salt thereof as an effective ingredient.

2. The pharmaceutical composition according to claim 1, characterized in that The neurodegenerative disease is selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, Creutzfeldt-Jakob disease, Lou Gehrig's disease, spinal muscular atrophy, Niemann-Pick disease, synucleinopathies, and dementia.

3. The pharmaceutical composition according to claim 1, characterized in that A pharmaceutically acceptable excipient is also included.

4. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition protects nerve cells from neurotoxicity.

5. The pharmaceutical composition according to claim 4, characterized in that The pharmaceutical composition protects nerve cells in the following ways: (i) inhibiting the death of dopaminergic neurons; (ii) inhibiting inflammation-mediated neuronal damage or neuronal degeneration; (iii) inhibiting the formation of α-synuclein Ser-129 phosphorylated aggregates; and / or (iv) inhibiting the abnormal increase in the number and / or activation of astrocytes.

6. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition (i) improves the exercise capacity of a subject to which the pharmaceutical composition is administered; and / or (ii) prolongs the life span of a subject to which the pharmaceutical composition is administered.

7. A food composition, characterized in that Contains fexofenadine.

8. The food composition according to claim 7, characterized in that The food is a health functional food for preventing or improving neurodegenerative diseases.

9. The food composition according to claim 8, characterized in that The neurodegenerative disease is selected from the group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, Creutzfeldt-Jakob disease, Lou Gehrig's disease, spinal muscular atrophy, Niemann-Pick disease, synucleinopathies, and dementia.

10. A composition for protecting nerve cells, characterized in that: Contains fexofenadine as the active ingredient.

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