Compositions for treating neurodegenerative diseases comprising beta-blockers and cholinesterase inhibitors - Patent Application 20070233333

Abstract

The present invention relates to a composition for treating neurodegenerative diseases, comprising a β-blocker and a cholinesterase inhibitor, and more specifically, to a composition that exhibits neuroinflammation alleviation, neuronal differentiation promotion and antioxidant effects, and exhibits a synergistic effect on neurodegenerative diseases.

Description

[Technical field]

[0001] This application claims priority to Korean Patent Application No. 10-2022-0014784, filed on February 4, 2022, the entire specification of which is incorporated herein by reference.

[0002] The present invention relates to a composition for treating neurodegenerative diseases, comprising a β-blocker and a cholinesterase inhibitor, and more specifically, to a composition that alleviates neuroinflammation, promotes neuronal differentiation, and exhibits antioxidant effects, and exhibits a synergistic effect on neurodegenerative diseases. [Background technology]

[0003] Neurodegenerative diseases are degenerative diseases that occur in the brain or spinal cord due to aging, and are caused by the gradual loss of structure and function of specific brain cell groups in the brain and spinal cord due to unknown causes, genetic defects, or environmental factors. Such neurodegenerative diseases include Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), multiple sclerosis (MS), and amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig's disease.

[0004] The causes of Alzheimer's disease are known to be a decrease in neurotransmission due to a decrease in acetylcholine concentration, the occurrence of neuroinflammation due to cytokines, cytotoxicity due to aggregation of amyloid beta and hyperphosphorylated tau protein, and neuronal death due to active oxygen caused by oxidative stress. Such death of neural stem cells has been pointed out as a fundamental pathology common to several brain diseases, and in-depth research is being conducted into the factors that cause cell death and the molecular mechanisms. However, the reality is that previously developed degrading enzyme agents and broad-spectrum cell death inhibitors have side effects and reduced clinical efficacy.

[0005] ALS is a fatal neurodegenerative disease that results in the selective loss of motor neurons due to cell death in the spinal cord. Approximately 10-20% of patients show a genetic pattern (familial ALS; fALS), while the rest are classified as sporadic ALS (sALS). Several genes are known at the ALS-associated locus in familial ALS, among which SOD1 is the first gene identified in fALS. It is speculated that fALS-associated genes also act in the development of sALS, but to date the exact cause of sALS remains unclear. Furthermore, ALS is classified into typical ALS, ALS with dementia, and atypical ALS according to the clinical symptoms. Indeed, mutations in SOD1 cause typical ALS, and C9orf72 is associated with ALS with dementia.

[0006] One of the key features of ALS is that it is a progressive disease, and neuronal death propagates through neural connections. In fact, Alzheimer's disease and Parkinson's disease also show similar phenotypes. Regarding the above features, a prion-like propagation model has been proposed, which states that misfolded proteins transform normal proteins into abnormal proteins. In fact, mutated amyloid beta (Aβ) can change normal Aβ into abnormal Aβ. Recently, it has been reported that mutated or misfolded SOD1 can also be secreted and propagated during the progression of the disease. However, there has been little research on SOD1 aggregation and misfolding inhibitors as a target for ALS disease therapeutics. Summary of the Invention [Problem to be solved by the invention]

[0007] Therefore, the inventors conducted extensive research to develop a combination of therapeutic agents that would have a synergistic effect in the treatment of neurodegenerative diseases from among known substances whose safety has been confirmed in clinical trials. As a result, they discovered that a combination preparation containing a β-blocker and a cholinesterase inhibitor exhibits significantly improved effects in alleviating neuroinflammation and promoting neuronal differentiation, compared to each drug alone, and thus completed the present invention.

[0008] Therefore, an object of the present invention is to provide a pharmaceutical composition for preventing or treating a neurodegenerative disease, which comprises a β-blocker and a cholinesterase inhibitor as active ingredients.

[0009] An object of the present invention is to provide a pharmaceutical composition for preventing or treating a neurodegenerative disease, comprising a β-blocker and a cholinesterase inhibitor.

[0010] An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of a neurodegenerative disease consisting essentially of a beta blocker and a cholinesterase inhibitor.

[0011] Another object of the present invention is to provide a food composition for preventing or ameliorating a neurodegenerative disease, which contains a β-blocker and a cholinesterase inhibitor as active ingredients.

[0012] Another object of the present invention is to provide a food composition for preventing or ameliorating a neurodegenerative disease, comprising a β-blocker and a cholinesterase inhibitor.

[0013] Another object of the present invention is to provide a food composition for preventing or ameliorating a neurodegenerative disease, which consists essentially of a beta blocker and a cholinesterase inhibitor.

[0014] The object of the present invention is to provide the use of a beta blocker and a cholinesterase inhibitor for the preparation of a pharmaceutical composition for the treatment of a neurodegenerative disease.

[0015] An object of the present invention is to provide a method for treating a neurodegenerative disease, comprising administering to an individual in need thereof an effective amount of a composition comprising a beta blocker and a cholinesterase inhibitor as active ingredients. [Means for solving the problem]

[0016] In order to achieve the above-mentioned object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating a neurodegenerative disease, which comprises a β-blocker and a cholinesterase inhibitor as active ingredients.

[0017] Furthermore, the present invention provides a pharmaceutical composition for preventing or treating a neurodegenerative disease, comprising a β-blocker and a cholinesterase inhibitor.

[0018] Furthermore, the present invention provides a pharmaceutical composition for preventing or treating a neurodegenerative disease, consisting essentially of a beta blocker and a cholinesterase inhibitor.

[0019] In order to achieve another object of the present invention, the present invention provides a food composition for preventing or improving a neurodegenerative disease, which contains a β-blocker and a cholinesterase inhibitor as active ingredients.

[0020] Furthermore, the present invention provides a food composition for preventing or ameliorating a neurodegenerative disease, comprising a β-blocker and a cholinesterase inhibitor.

[0021] Furthermore, the present invention provides a food composition for preventing or ameliorating a neurodegenerative disease, which consists essentially of a β-blocker and a cholinesterase inhibitor.

[0022] In order to achieve another object of the present invention, the present invention provides the use of a beta blocker and a cholinesterase inhibitor for the preparation of a pharmaceutical composition for the treatment of a neurodegenerative disease.

[0023] In order to achieve another object of the present invention, the present invention provides a method for treating a neurodegenerative disease, comprising administering to an individual in need thereof an effective amount of a composition comprising a β-blocker and a cholinesterase inhibitor as active ingredients.

[0024] As used herein, the term "treatment" or "treating" is defined as the application or administration of a therapeutic agent, i.e., a compound of the present invention (alone or in combination with other pharmaceutical preparations), to a patient or to an isolated tissue or cell line for the purpose of treating, curing, mitigating, alleviating, altering, resolving, ameliorating, improving or affecting HBV infection, symptoms of HBV infection, or the likelihood of developing HBV infection in a patient (e.g., diagnostic or therapeutic applications). Such treatments may be tailored and modified based on knowledge gained, particularly from the field of genomic drug discovery.

[0025] The term "prevent" or "prevention" as used herein means that there is no progression of a disorder or disease if nothing has happened, or that there is no further progression of a disorder or disease if there is already progression of a disorder or disease. Additionally, what is considered is the ability of one to prevent some or all of the symptoms associated with a disorder or disease.

[0026] The term "patient", "individual" or "subject" as used herein refers to a human or non-human mammal. For example, non-human mammals include livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. Preferably, the patient, subject or individual is a human.

[0027] As used herein, the terms "effective amount", "pharmaceutical effective amount" and "therapeutically effective amount" refer to a non-toxic sufficient amount of a formulation to produce a desired biological result. Said result may be a reduction and / or attenuation of a signal, symptom, or cause of a disease, or any other desired change in a biological system. For any individual, the appropriate therapeutic amount can be determined by one of ordinary skill in the art using routine experimentation.

[0028] As used herein, the term "pharmaceutical acceptable" refers to a substance, such as a carrier or diluent, that does not interfere with the biological activity or properties of the compound and is relatively non-toxic, i.e., the substance may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0029] The term "pharmaceutical acceptable salts" as used herein refers to salts of compounds prepared and administered from pharmaceutically acceptable non-toxic acids, including inorganic acids, organic acids, solvates, hydrates or clathrates thereof. Examples of such inorganic acids include hydrochloric acid, bromic acid, iodic acid, nitric acid, sulfuric acid, phosphoric acid, acetic acid, hexafluorophosphoric acid, citric acid, gluconic acid, benzoic acid, propionic acid, butyric acid, sulfosalicylic acid, maleic acid, lauric acid, malic acid, fumaric acid, succinic acid, tartaric acid, amsonic acid, palmic acid, p-toluenesulfonic acid and mesylic acid. Suitable organic acids can be selected, for example, from aliphatic, aromatic, carboxylic and sulfonic organic acids, such as formic acid, acetic acid, propionic acid, succinic acid, camposulfonic acid, citric acid, fumaric acid, gluconic acid, isethionic acid, lactic acid, malic acid, mucic acid, tartaric acid, paratoluenesulfonic acid, glycolic acid, glucuronic acid, maleic acid, furoic acid, glutamic acid, benzoic acid, atranilic acid, salicylic acid, phenylacetic acid, mandelic acid, enfonic acid (palm acid), methanesulfonic acid, ethanesulfonic acid, pantothenic acid, benzenesulfonic acid (besylate), stearic acid, sulfanilic acid, alginic acid, galacturonic acid, etc. Additionally, pharma- ceutically acceptable salts are non-limiting examples and include alkaline earth metal salts (e.g., calcium or magnesium), alkali metal salts (e.g., sodium-dependent or potassium) and ammonium salts.

[0030] The term "pharmaceutical acceptable carrier" as used in the present invention means a pharmaceutically acceptable substance, composition or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, additive, thickener, solvent or encapsulating material, associated with the transport of the compound useful in the present invention so that it can perform its intended function in or on a patient. Typically, such compositions are transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense that it can be used with other components of the formula containing the compound useful in the present invention and is not damaging to the patient. Some examples of substances which can act as pharma- ceutically acceptable carriers include: sugars used in pharmaceutical preparations such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and derivatives such as sodium carboxymethylcellulose, ethylcellulose, cellulose acetate; powdered tragacanth; malt; gelatin; talc; additives such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, polyethylene glycol; esters such as ethyl oleate, ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic solutions; Ringer's solution; ethyl alcohol; phosphate buffers; and other non-toxic compatible substances. In addition, the "pharmaceutically acceptable carrier" used in the present invention includes any coating agent, antiviral and antibacterial agents, absorption delaying agents, and the like, that is compatible with the activity of the compound useful in the present invention and is physiologically acceptable to the patient. In addition, supplementary active compounds can be incorporated into the composition. In addition, the "pharmaceutically acceptable carrier" can further include pharmaceutically acceptable salts of the compound useful in the present invention. Other additional components that can be included in the pharmaceutical compositions used in the practice of the present invention are known in the art.

[0031] The term "composition" or "pharmaceutical composition" as used in the present invention refers to a mixture of at least one compound useful in the present invention and a pharma- ceutical acceptable carrier.The pharmaceutical composition facilitates the administration of the compound to a patient or subject.There are various techniques in the art for administering the compound, including, but not limited to, intravenous injection, oral, aerosol, parenteral, ocular, pulmonary, and topical administration.

[0032] The term "combination therapy" or "in combination" as used herein refers to the administration of two or more therapeutic agents to treat the conditions or disorders described herein (i.e., metabolic and fibrotic diseases). Such administration includes co-administration of these therapeutic agents in a substantially simultaneous manner, e.g., in a single capsule having a fixed ratio of active ingredients. Alternatively, such administration includes co-administration in multiple or separate containers (e.g., capsules, powders, and liquids) for each active ingredient. The powders and / or liquids can be reconstituted or diluted to the desired dose before administration. Additionally, such administration also includes the sequential use of each type of therapeutic agent at about the same time or at different times. In either case, the treatment regimen will provide the beneficial effects of the drug combination in treating the conditions or disorders described herein.

[0033] Combination therapy can provide a "synergistic effect" and demonstrate "synergistic". That is, the effect achieved when the active ingredients are used together is greater than the sum of the effects produced by using the compounds individually. Synergistic effects can be obtained when the active ingredients are (1) co-formulated and administered or delivered simultaneously as a combined unit dose formulation; (2) delivered alternately or in parallel as individual formulations; or (3) provided by some other scheme. When delivered in alternation therapy, synergistic effects can be obtained when the compounds are administered or delivered sequentially, e.g., by different injections from each other in separate syringes. Generally, during alternation therapy, an effective dose of each active ingredient is administered sequentially, i.e., consecutively, whereas in combination therapy, effective doses of two or more active ingredients are administered together. Synergistic effect, as used herein, refers to the action of two therapeutic agents that produces an effect greater than the simple sum of the effects of each drug administered alone. Synergistic effect can be calculated using any suitable method, such as, for example, the equation of the sigmoid Emax model, the Loewe summation equation, etc. Each of the above equations can be applied to experimental data to generate corresponding graphs useful for evaluating the effect of drug combinations. The corresponding graphs associated with the above equations are the concentration-effect curve, the isobologram curve, and the combination index curve, respectively.

[0034] The present invention will be described in detail below.

[0035] The present invention provides a pharmaceutical composition for preventing or treating a neurodegenerative disease, which comprises a β-blocker and a cholinesterase inhibitor as active ingredients.

[0036] In the present invention, "beta blocker" refers to a beta receptor blocker, a beta adrenergic receptor blocker, a beta blocker, a beta blocker or a beta adrenergic receptor blocker, or a natural or man-made compound that inhibits the binding of agonists that bind to all types of beta adrenergic receptors (beta-1, beta-2, beta-3 or other).

[0037] Beta-blockers are known to exert positive effects on the cardiovascular system, primarily by blocking cardioselective beta-1 receptors. Many different beta-blockers are approved for the treatment of cardiovascular disease in humans. Through their inotrope and chronotropic inhibitory effects, beta-blockers directly improve hemodynamic cardiac workload economy. Beta-blockers are used in humans for the treatment of hypertension, coronary artery disease (CAD) and prevention of heart attacks, as well as for the treatment of stable chronic heart failure with limited systolic function, arrhythmias, and hypermobility heart syndrome.

[0038] In the present invention, the β-blocker may be selected from the group consisting of nebivolol, acebutolol, alprenolol, atenolol, betaxolol, bisoprolol, bucindolol, carteolol, carvedilol, celiprolol, esmolol, labetalol, levobunolol, medroxalol, mepindolol, metipranolol, metoprolol, nadolol, oxyprenolol, penbutolol, pindolol, propranolol, sotalol and timolol, but is not limited thereto, and most preferably may be nebivolol.

[0039] Nebivolol is a compound having the structure of the following formula 1, and is beta-blocking, but is different from other classical beta-blockers in that it has high selectivity for beta1-adrenergic receptors and retains the vasodilatory effect associated with its effect on endothelial nitric oxide. Nebivolol is thought to increase the concentration of nitric oxide via the L-arginine-nitric oxide pathway in the vascular endothelium and has been found to improve endothelial dysfunction and vascular elasticity. Nebivolol has been found to be beneficial in the treatment of cardiovascular diseases such as hypertension, congestive heart failure, arteriosclerosis and endothelial dysfunction.

[0040] [ka]

[0041] The "cholinesterase inhibitor" in the present invention refers to a compound that inhibits the enzymatic degradation of the neurotransmitter acetylcholine, thus increasing the duration and level of acetylcholine in the synaptic cleft. Two enzymes are primarily responsible for the degradation of acetylcholine, acetylcholinesterase and butyrylcholinesterase. "Cholinesterase inhibitor" includes substances that inhibit or otherwise reduce the action of one or both of these enzymes.

[0042] In the compositions and methods of the present invention, the cholinesterase inhibitor is considered to be pharmacologic. As used herein, "pharmacologic" means that the cholinesterase inhibitor is therapeutically useful in humans. Thus, the term excludes cholinesterase inhibitors used as pesticides, such as aldicarb (2-methyl-2-methylthiopropionaldehyde-O-methylcarbamoyloxime), carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranylmethylcarbamate), carbaryl (1-naphthylmethylcarbamate), and cholinesterase inhibitors that are lethal to humans and are used as chemical weapons, such as sarin (2-(fluoro-methylphosphoryl)oxypropane), VX (S-[2-(diisopropylamino)ethyl]-O-ethylmethylphosphonothioate), and soman (3-(fluoro-methyl-phosphoryl)oxy-2,2-dimethyl-butane). Most cholinesterase inhibitors that are pesticides and chemical warfare agents are quasi-reversible or irreversible. Most pharmacologically effective cholinesterase inhibitors are reversible.

[0043] Pharmaceutically effective cholinesterase inhibitors are widely known in the art. Examples include, but are not limited to, 7-methoxytacrine, albamerin, ambenonium, ansecrine, arecoline, cevimeline, citicoline, demacarium, donepezil, edrophonium, eptastigmine, fasciculin, heptylphysostigmine, huperzine A and analogs thereof, icopezil, ipidacrine, linopyridine, metrifonate, miramelin, neostigmine, nomeostigmine, pyridostigmine, norpyridostigmine, tacrine, physostigmine, rivastigmine, sabcomeline, slonacrine, tacrine analogs, tacrine, talsaclidine, velnacrine, xanomeline, diflosillone, itopride, acotiamide, huperzine, galantamine, salts thereof, and the like, and most preferably, the cholinesterase inhibitor can be donepezil.

[0044] Donepezil is an acetylcholinesterase (AChE) inhibitor with the structure of the following formula 2, and is used to treat mild to severe dementia of Alzheimer's disease. In Alzheimer's disease, where cholinergic nervous system disorders in the brain have been reported, donepezil increases acetylcholine in the brain and activates cholinergic nerves in the brain. Donepezil preparations currently on the market are in tablet form, and are prescribed to Alzheimer's disease patients in oral form.

[0045] [ka]

[0046] According to one embodiment of the present invention, a combination composition containing nebivolol and donepezil has been confirmed to exhibit synergistic effects in alleviating neuroinflammation, neuronal differentiation, and antioxidant effects compared to each single formulation, and has been found to be effective in preventing or treating neurodegenerative diseases.

[0047] Therefore, the composition of the present invention containing a β-blocker and a cholinesterase inhibitor can show improved effects at lower doses compared to when each drug is used alone, thereby reducing side effects and improving the therapeutic effect on brain diseases, i.e., has the advantage of a synergic effect.

[0048] The Bliss independence combined response C for two single compounds with effects A and B is C = A + BA * B, where each effect is expressed as a fractional inhibition value between 0 and 1 (see Bliss (1939) Annals of Applied Biology). The Bliss value, defined as the difference between the experimental response and the calculated Bliss independence value, indicates whether the effects of the two components in combination are additive or synergistic.

[0049] A Bliss value of zero (0) is considered additive. The term "additive" means that the result of combining two targeted agents is the sum of each of the individual drugs.

[0050] The term "synergy" or "synergy" is used to mean that the response of the combination of two preparations is greater than the sum of the responses of each of the individual preparations. More specifically, in an in vitro context, one measure of synergy is known as "Bliss synergy". Bliss synergy means "exceeding the Bliss independent value" as determined by the Bliss value mentioned above. If the Bliss value is greater than zero (0), it is considered an indication of synergy. Of course, the concept of "synergy" used in the present invention also includes in vitro synergy measured by additional and / or alternative methods.

[0051] The in vitro biological effect of the combination of a beta blocker and a cholinesterase inhibitor of the present invention, including but not limited to an anti-inflammatory effect, that is greater than or equal to the sum of the individual components of the combination can be correlated with the Bliss value. Furthermore, "synergy," which includes when a combination of components used in the present invention exhibits activity equal to or greater than the sum of the individual components, can be measured by additional and / or alternative methods.

[0052] In one aspect of the invention, the composition of the invention is for treating a neurodegenerative disease, and can combine an effective amount of a beta-blocker, or a pharma- ceutical effective salt, derivative, or metabolite thereof, with an effective amount of a cholinesterase inhibitor, or a pharma- ceutical effective salt thereof, in an amount sufficient to achieve a synergistic effect.

[0053] In one embodiment of the present invention, the β-blocker and the cholinesterase inhibitor may be included in a molar ratio of 1:0.1-20, preferably in a molar ratio of 1:0.1-10, more preferably in a molar ratio of 1:0.1-5, more preferably in a molar ratio of 1:0.1-3, even more preferably in a molar ratio of 1:0.3-3, and most preferably in a molar ratio of 1:0.5-2.

[0054] In the present invention, the neurodegenerative disease may be selected from the group consisting of Alzheimer's disease, Parkinson's disease, dementia, cognitive impairment, progressive supranuclear palsy, multiple system atrophy, olivopontocerebellar atrophy (OPCA), Shy-Drager syndrome, striatonigral degeneration, Huntington's disease, amyotrophic lateral sclerosis (ALS), essential tremor, corticobasal degeneration, diffuse Lewy body disease, Perkins-ALS-dementia complex, Niemann-Pick disease, and Pick's disease, but is not limited thereto.

[0055] The cognitive dysfunction can include, without limitation, any disease that is closely related to aging, but is different from the normal aging process, and that rapidly causes abnormal neuronal death in a part of the nervous system or the whole brain, resulting in loss of brain or spinal cord function and reduced cognitive ability.Non-limiting examples of cognitive dysfunction can include mild cognitive impairment, Alzheimer's disease, frontotemporal dementia, Lewy body disease, corticobasal degeneration, learning disability, agnosia, forgetfulness, aphasia, apraxia, and delirium.

[0056] In the composition of the present invention, the β-blocker and the cholinesterase inhibitor can be administered simultaneously, separately or sequentially, and the β-blocker and the cholinesterase inhibitor can exert preventive or therapeutic activity against brain diseases by synergistic action in the body.

[0057] Generally, beta-blockers and cholinesterase inhibitors are administered simultaneously as a composition. Even if the active ingredients are administered to the human body at different times, the individually administered active ingredients act simultaneously in the body, thereby achieving the same level of therapeutic activity.

[0058] Specifically, "simultaneous administration" means that the above two active ingredients are administered together via the same administration route, or administered at substantially the same time (e.g., administration time interval of 15 minutes or less) via the same or different administration routes. Separate administration means that the two active ingredients are administered via the same or different administration routes with a certain time interval (e.g., 3 days apart). Sequential administration means that both active ingredients are administered via the same or different administration routes with a certain order rule according to the disease state of the patient.

[0059] The administration route can be oral or parenteral. Parenteral administration methods can be, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, percutaneous, subcutaneous, intraneuronal, intraventricular (subventricular region), intracerebrovascular, intraperitoneal, intranasal, intestinal, topical, sublingual, or rectal administration.

[0060] The pharmaceutical composition according to the present invention may contain only a pharma- ceutical effective amount of a β-blocker and a cholinesterase inhibitor, or may additionally contain a pharma- ceutical acceptable carrier. The term "pharma-ceutical effective amount" refers to an amount that shows a greater response than a negative control group, and preferably means an amount sufficient to show the effects of extending life span, improving mobility, suppressing neuroinflammation, suppressing neuronal death, and promoting neuronal differentiation by administering the two active ingredients in combination in the treatment or prevention of brain diseases.

[0061] The pharmaceutical composition of the present invention can be formulated in various ways according to the administration route with a pharma- ceutically acceptable carrier in a manner known in the art to show the synergistic effect of combining a cholinesterase inhibitor and an antioxidant. "Pharmaceutically acceptable" refers to a non-toxic composition that is physiologically acceptable, does not inhibit the action of the active ingredient when administered to humans, and does not usually cause allergic reactions or similar reactions such as gastrointestinal disorders or dizziness. The carrier includes any kind of solvent, dispersion medium, oil-in-water or water-in-oil emulsion, aqueous composition, liposome, microbeads and microsome. The pharma- ceutically acceptable carriers contained in the composition are those commonly used in formulations, and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, etc. Other pharma-ceutically acceptable carriers can be referred to those known in the art.

[0062] In addition to the ingredients, the pharmaceutical composition may further contain lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. Specifically, for oral administration, binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, dyes or flavors, etc. may be used, for injections, buffers, preservatives, soothing agents, solubilizers, isotonic agents, stabilizers, etc. may be mixed and used, and for topical administration, bases, excipients, lubricants, preservatives, etc. may be used.

[0063] Furthermore, the composition of the present invention can be used in the form of a general pharmaceutical preparation. Parenteral preparations can be prepared in the form of a sterile aqueous solution, non-aqueous solvent, suspension, emulsion or lyophilized preparation, injection, transdermal injection, nasal inhalation, etc., and oral administration can be prepared in the form of a tablet, troche, capsule, elixir, suspension, syrup or wafer. Injections can be prepared in the form of a unit dose ampoule or multiple doses. Injections must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. Examples of suitable carriers for injections include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), mixtures thereof, and / or vegetable oils. More preferably, suitable carriers include Hank's solution, Ringer's solution, PBS (phosphate buffered saline) containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. To protect the injectable preparation from microbial contamination, it may further contain various antibacterial and antifungal agents, such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In addition, in most cases, the injectable preparation may further contain an isotonic agent, such as sugar or sodium chloride.

[0064] Furthermore, the pharmaceutical composition of the present invention can be administered by any device that allows the active substance to be transferred to the target site. Preferred administration methods and formulations include intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, or drip injections. The injections can be prepared using aqueous solvents such as physiological saline or Ringer's solution, non-aqueous solvents such as vegetable oils, higher fatty acid esters (e.g., ethyl oleate, etc.), alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, or glycerin, etc.), and may contain pharmaceutical carriers such as stabilizers for preventing deterioration (e.g., ascorbic acid, sodium hydrogen sulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for adjusting pH, and preservatives for preventing microbial growth (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.). The method of treating or preventing neurodegenerative diseases using the composition of the present invention includes administering an effective amount (pharmaceutical effective amount) of the therapeutic composition of the present invention to an individual in need thereof. The above-mentioned pharmacologic effective amount can be easily determined by a person skilled in the art according to factors well known in the medical field, such as the type of disease, the patient's age, weight, health, sex, sensitivity of the patient to the drug, administration route, administration method, number of administrations, treatment period, and drugs used in combination or concomitantly.

[0065] Moreover, the pharmaceutical compositions of the present invention can be formulated using methods known in the art so as to provide quick, sustained or delayed release of the active ingredient after administration to a mammal.

[0066] The total effective amount of the composition of the present invention can be administered to a patient in a single dose or in a fractionated treatment protocol in which multiple doses are administered over a long period of time. The pharmaceutical composition of the present invention can vary the content of the active ingredient depending on the severity of the disease. Preferably, the total preferred dose of the pharmaceutical composition of the present invention can be about 0.01 μg to 10,000 mg per kg of patient body weight per day, most preferably 0.1 μg to 500 mg. However, the dosage of the pharmaceutical composition is determined by taking into consideration various factors such as the age, body weight, health condition, sex, severity of the disease, diet, and excretion rate of the patient, as well as the formulation method, administration route, and number of treatments. Therefore, taking these points into consideration, a person having conventional knowledge in the art would be able to determine the appropriate effective dose of the composition of the present invention. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route, and administration method, as long as it shows the effect of the present invention.

[0067] The composition of the present invention may be formulated so that the components of the β-blocker and the cholinesterase inhibitor are simultaneously contained in one formulation depending on the administration method and route, or each of the components may be formulated separately and contained in one package according to the daily or once-a-day dosage unit. The formulations of the β-blocker and the cholinesterase inhibitor separately formulated may or may not be the same. The specific formulation method of the pharmaceutical composition of the present invention and the pharmacologic acceptable carrier that can be contained in the formulation are as described above for the pharmaceutical composition and are known in the art.

[0068] The present invention also provides a food composition for preventing or ameliorating a neurodegenerative disease, which contains a β-blocker and a cholinesterase inhibitor as active ingredients.

[0069] The food composition of the present invention includes all forms of functional foods, nutritional supplements, health foods, food additives, etc., and is intended for consumption by humans or animals, including livestock. This type of food composition can be prepared in various forms according to conventional methods known in the art.

[0070] This type of food composition can be prepared in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meat and processed foods thereof (e.g., ham, sausage, corned beef, etc.), breads and noodles (e.g., udon, soba, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, candies, dairy products (e.g., butter, cheese, etc.), edible plant foods, margarine, vegetable proteins, retort foods, frozen foods, various seasonings (e.g., miso, soy sauce, sauces, etc.), etc., which can be prepared by adding the above-mentioned complex preparation. Nutritional supplements include, but are not limited to, capsules, tablets, pills, etc., which can be prepared by adding the above-mentioned complex preparation. In addition, the specific health food is not limited thereto, but for example, the complex preparation itself can be prepared in the form of tea, juice, or drink, and can be liquefied, granulated, encapsulated, or powdered for consumption (health drink). In addition, in order to use the complex preparation as a food additive, it can be prepared in the form of powder or concentrated liquid. In addition, the complex preparation can be mixed with a known active ingredient known to have an effect of improving neurodegenerative diseases to prepare a composition.

[0071] When the food composition of the present invention is used as a health drink composition, the health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients, as in ordinary drinks. The natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener may be a natural sweetener such as thaumatin or stevia extract; or an artificial sweetener such as saccharin or aspartame. The proportion of the natural carbohydrates is usually about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the composition of the present invention.

[0072] When the composite preparation according to the present invention is contained as an active ingredient in a food composition, the amount is not particularly limited to an amount effective for achieving symptom improvement, but is preferably 0.01 to 100% by weight based on the total weight of the entire composition. The food composition of the present invention can be produced by mixing the composite preparation with other active ingredients known to have an effect of improving neurodegenerative diseases in combination.

[0073] In addition to the above, when the food composition of the present invention is used as a health food, it may contain various nutrients, vitamins, electrolytes, flavor improvers, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol or carbonation agents. In addition, the health food of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice beverage or vegetable beverage. These ingredients may be used independently or in mixture. The proportion of such additives is not very important, but is generally selected from the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

[0074] The present invention provides the use of a beta blocker and a cholinesterase inhibitor for the manufacture of a pharmaceutical composition for treating a neurodegenerative disease.

[0075] The present invention provides a method for treating a neurodegenerative disease, comprising administering to an individual in need thereof an effective amount of a composition comprising a β-blocker and a cholinesterase inhibitor as active ingredients.

[0076] The above-mentioned "effective amount" of the present invention refers to an amount that, when administered to an individual, shows an effect of improving, treating, detecting, diagnosing a neurodegenerative disease or a neurodegenerative disease, or suppressing or reducing the disease, and the "individual" may be an animal, preferably a mammal, particularly an animal including a human, and may be a cell, tissue, organ, etc. derived from an animal. The individual may be a patient in need of the effect.

[0077] The "treatment" of the present invention refers generally to improving a neurodegenerative disease or symptoms caused by the disease, which may include curing, substantially preventing, or ameliorating the condition of the disease, including, but not limited to, reducing, curing, or preventing one or most symptoms resulting from the disease.

[0078] The term "comprising" in this specification is used interchangeably with "including" or "characterized by" and does not exclude additional components or method steps not specifically mentioned in the compositions or methods according to the present invention. The term "consisting of" means excluding additional elements, steps, or ingredients not otherwise described. The term "essentially consisting of" means that the scope of a composition or method may include, in addition to the materials or steps described, materials or steps that do not substantially affect its basic properties. Effect of the Invention

[0079] The composition according to the present invention shows enhanced preventive and therapeutic effects for neurodegenerative diseases by administering a β-blocker and a cholinesterase inhibitor in combination compared to administration of each drug alone, and has the effect of reducing side effects that may be caused by excessive administration or long-term administration of each drug. [Brief description of the drawings]

[0080] [Figure 1a] Figures 1a to 1e show the results of evaluating the anti-inflammatory effect by measuring the NO concentration generated after treating microglia (BV2 cells) with nebivolol alone, donepezil alone, or a combination of nebivolol and donepezil (NDC-011) together with LPS (lipopolysaccharide), an inflammatory response inducer. [Figure 1b] Figures 1a to 1e show the results of evaluating the anti-inflammatory effect by measuring the NO concentration generated after treating microglia (BV2 cells) with nebivolol alone, donepezil alone, or a combination of nebivolol and donepezil (NDC-011) together with LPS (lipopolysaccharide), an inflammatory response inducer. [Figure 1c] Figures 1a to 1e show the results of evaluating the anti-inflammatory effect by measuring the NO concentration generated after treating microglia (BV2 cells) with nebivolol alone, donepezil alone, or a combination of nebivolol and donepezil (NDC-011) together with LPS (lipopolysaccharide), an inflammatory response inducer. [Figure 1d] Figures 1a to 1e show the results of evaluating the anti-inflammatory effect by measuring the NO concentration generated after treating microglia (BV2 cells) with nebivolol alone, donepezil alone, or a combination of nebivolol and donepezil (NDC-011) together with LPS (lipopolysaccharide), an inflammatory response inducer. [Figure 1e]Figures 1a to 1e show the results of evaluating the anti-inflammatory effect by measuring the NO concentration generated after treating microglia (BV2 cells) with nebivolol alone, donepezil alone, or a combination of nebivolol and donepezil (NDC-011) together with LPS (lipopolysaccharide), an inflammatory response inducer. [Figure 2a] Figures 2a to 2f show the results of evaluating the differentiation-promoting effect of drugs by assessing the expression level of HB9, a motor neuron marker, after culturing neural progenitor cells (ReNcell VM) under differentiation conditions in which they were treated with nebivolol alone, donepezil alone, or in combination (NDC-011). [Figure 2b] Figures 2a to 2f show the results of evaluating the differentiation-promoting effect of drugs by assessing the expression level of HB9, a motor neuron marker, after culturing neural progenitor cells (ReNcell VM) under differentiation conditions in which they were treated with nebivolol alone, donepezil alone, or in combination (NDC-011). [Figure 2c] Figures 2a to 2f show the results of evaluating the differentiation-promoting effect of drugs by assessing the expression level of HB9, a motor neuron marker, after culturing neural progenitor cells (ReNcell VM) under differentiation conditions in which they were treated with nebivolol alone, donepezil alone, or in combination (NDC-011). [Figure 2d] Figures 2a to 2f show the results of evaluating the differentiation-promoting effect of drugs by assessing the expression level of HB9, a motor neuron marker, after culturing neural progenitor cells (ReNcell VM) under differentiation conditions in which they were treated with nebivolol alone, donepezil alone, or in combination (NDC-011). [Figure 2e] Figures 2a to 2f show the results of evaluating the differentiation-promoting effect of drugs by assessing the expression level of HB9, a motor neuron marker, after culturing neural progenitor cells (ReNcell VM) under differentiation conditions in which they were treated with nebivolol alone, donepezil alone, or in combination (NDC-011). [Figure 2f] Figures 2a to 2f show the results of evaluating the differentiation-promoting effect of drugs by assessing the expression level of HB9, a motor neuron marker, after culturing neural progenitor cells (ReNcell VM) under differentiation conditions in which they were treated with nebivolol alone, donepezil alone, or in combination (NDC-011). [Diagram 3] Figure 3 shows the results of treating neuroblastoma cells (SH-SY5Y) with nebivolol alone, donepezil alone, or the combination (NDC-011), inducing oxidative stress through treatment with hydrogen peroxide, and then measuring the amount of reactive oxygen species (ROS) generated to evaluate the antioxidant effect of the drugs. [Figure 4] Figure 4 shows the results of the evaluation of the animals' motor abilities by a Rotarod latency test at weekly intervals after oral administration of edaravone or a combination of nebivolol and donepezil (NDC-011) to a 70-day-old ALS animal model. [Diagram 5] Figure 5 shows the results of administering edaravone or a combination of nebivolol and donepezil (NDC-011) to a 70-day-old ALS animal model for 28 days, sampling spinal cord tissue, and comparing the number of motor neurons in the spinal cord tissue via Nissl staining. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0081] The present invention will now be described in more detail with reference to the following examples, although it should be noted that the following examples are intended to illustrate the present invention and are not intended to limit the present invention.

[0082] Example 1: Anti-inflammatory effect of nebivolol + donepezil complex (in vitro) In the present invention, microglia (BV2 cells) were treated with LPS to induce an inflammatory response, and the concentration of nitric oxide (NO) generated after treatment with nebivolol alone, donepezil alone, or a combination of nebivolol and donepezil was measured to confirm the neuroinflammation-reducing effects of the drugs.

[0083] Microglia were treated with LPS to induce an inflammatory response, and NO generation was measured after treatment with nebivolol alone, donepezil alone, or the combination of nebivolol and donepezil. As a result, as shown in Figures 1a-1e, a significant decrease in NO was observed in the drug-treated groups compared to the LPS control group, and the reduction effect was greatest in the nebivolol + donepezil combination treatment group. In addition, the Bliss independence model was used to calculate the predicted combination treatment effect from the single treatment effect of both drugs, and the actual combination treatment effect of the two drugs was higher than that of the single treatment effect of both drugs, confirming the synergistic effect of the two drugs. (*P <0.05, **P <0.01, ***P <0.001, ****P <0.0001, one-way ANOVA)

[0084] Example 2: Effect of nebivolol + donepezil complex on promoting differentiation of neural progenitor cells (in vitro) Immunostaining was performed on ReNcell VM (immortalized human neural progenitor cell line), a neural progenitor cell that can differentiate into neurons and glial cells, using an antibody (anti-HB9 antibody) that can selectively bind to motor neurons after treatment with nebivolol alone, donepezil alone, or a combination of nebivolol and donepezil, and a CellTag probe that stains whole cells.The degree to which neural progenitor cells differentiate into motor neurons was measured by analyzing the HB9 positive signal and CellTag values ​​to confirm the number of motor neurons among all cells.

[0085] As a result, as shown in Figures 2a-2f, a significant increase in HB-9 expression was confirmed in the nebivolol alone and nebivolol + donepezil combined treatment groups compared to the control group. This increase was not observed in the low concentration (0.01 μM) nebivolol or donepezil alone treatment groups. The prediction results from the Bliss Independence Model also confirmed that the actual combined treatment effect was greater than the expected effect, and that nebivolol and donepezil exhibited a synergistic effect in promoting differentiation of neural progenitor cells. (* P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001, one-way ANOVA)

[0086] Example 3: Antioxidant effect of nebivolol + donepezil complex (in vitro) In the present invention, neuroblastoma cells (SH-SY5Y) were treated with nebivolol alone, donepezil alone, or a combination of nebivolol and donepezil, and then dichlorofluorescein (DCF), which emits a fluorescent signal proportional to the amount of reactive oxygen, and H2O2, an oxidative stress inducer, were treated to measure the amount of reactive oxygen species (ROS) generated, and the effect of the drugs in reducing reactive oxygen generation was confirmed.

[0087] As a result, as shown in Figure 3, ROS was significantly reduced only in the group treated with the combination of the two drugs compared to the control group, and no significant reduction was observed in the groups treated with either nebivolol or donepezil alone. In addition, the prediction results using the Bliss Independence Model confirmed that the actual combined treatment effect was greater than the expected effect, and a synergistic effect was observed in the antioxidant effect of the combination of nebivolol and donepezil (**P<0.01, * ***P<0.0001, one-way ANOVA).

[0088] Example 4: In vivo ALS treatment effect To verify the in vivo effect of the nebivolol and donepezil complex, SOD1G93A genetically modified (TG) mice, an ALS animal model, were used, and from the age of 70 days, edaravone (15 mg / kg), a commercially available ALS treatment drug, or the nebivolol and donepezil complex according to the present invention (NDC-011, nebivolol 5 mg / kg + donepezil 3 mg / kg) was orally administered once a day for 70 days. A rotarod latency test was performed immediately before the start of drug administration to evaluate the motor ability of each animal, and then rotarod latency tests were performed at 7-day intervals throughout the drug administration period to evaluate the motor ability of the ALS animals. As a result, the placebo group or the commercially available treatment edaravone group showed a significant decrease in motor ability compared to the normal control group from the age of 84 days, while the group administered with the complex according to the present invention showed a decrease in motor ability from the age of 123 days, confirming that the NDC-011 complex delays the time when the motor ability of ALS animals decreases by about 40 days (Figure 4).

[0089] To confirm the inhibitory effect of the NDC-011 complex on motor neuron cell death, edaravone or NDC-011 complex (nebivolol 5 mg / kg + donepezil 3 mg / kg) was administered to 70-day-old ALS animal models once daily for 28 days, after which the spinal cord tissue of TG mice was sampled and the motor neuron cells were stained with Nissl staining to confirm their number. As a result, the number of motor neurons in the placebo-administered group was significantly decreased compared to the normal control group, confirming that motor neuron cell death occurred in the ALS animal model. In the nebivolol and donepezil complex (NDC-011)-administered group, the number of motor neurons was significantly increased compared to the placebo-administered group or the edaravone-administered group, and the number of motor neurons was increased compared to the normal control group, confirming the inhibitory effect of the NDC-011 complex on motor neuron cell death and the increase in the number of motor neurons in the ALS animal model (Figure 5). [Industrial Applicability]

[0090] The composition according to the present invention shows enhanced preventive and therapeutic effects for neurodegenerative diseases by administering a β-blocker and a cholinesterase inhibitor in combination compared to administration of each drug alone, and has the effect of reducing side effects that may be caused by excessive administration or long-term administration of each drug, and can be very useful in the development of therapeutic agents for neurodegenerative diseases.

Claims

1. A pharmaceutical composition for preventing or treating a neurodegenerative disease, comprising a beta blocker and a cholinesterase inhibitor as active ingredients.

2. The pharmaceutical composition according to claim 1, characterized in that the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, dementia, cognitive impairment, progressive supranuclear palsy, multiple system atrophy, olivopontocerebellar atrophy (OPCA), Shy-Drager syndrome, striatonigral degeneration, Huntington's disease, amyotrophic lateral sclerosis (ALS), essential tremor, corticobasal degeneration, diffuse Lewy body disease, Parkins-ALS-dementia complex, Niemann-Pick disease, and Pick's disease.

3. 2. The pharmaceutical composition of claim 1, wherein the beta-blocker is selected from the group consisting of nebivolol, acebutolol, alprenolol, atenolol, betaxolol, bisoprolol, bucindolol, carteolol, carvedilol, celiprolol, esmolol, labetalol, levobunolol, medroxalol, mepindolol, metipranolol, metoprolol, nadolol, oxyprenolol, penbutolol, pindolol, propranolol, sotalol and timolol.

4. 2. The pharmaceutical composition according to claim 1, wherein the cholinesterase inhibitor is at least one selected from the group consisting of donepezil, 7-methoxytacrine, albamerin, ambenonium, ansecrine, arecoline, cevimeline, citicoline, demacarium, edrophonium, eptastigmine, fasciculin, heptylphysostigmine, huperzine A and its analogs, icopezil, ipidacrine, linopyridine, metrifonate, miramelin, neostigmine, nomeostigmine, pyridostigmine, norpyridostigmine, tacrine, physostigmine, rivastigmine, sabcomeline, slonacrine, tacrine analogs, tacrine, talsaclidine, velnacrine, xanomeline, itopride, acotiamide, huperzine, galantamine, and diflosilone, or a salt thereof.

5. The pharmaceutical composition according to claim 1, characterized in that the composition exhibits neuroinflammation alleviation, neuronal differentiation promotion and antioxidant effects.

6. The pharmaceutical composition according to claim 1, wherein the β-blocker and the cholinesterase inhibitor are co-administered in a molar ratio of 1:0.1-20.

7. 2. The pharmaceutical composition according to claim 1, wherein the beta-blocker and the cholinesterase inhibitor are co-administered simultaneously, separately or sequentially.

8. 2. The pharmaceutical composition according to claim 1, wherein the beta-blocker is nebivolol represented by the following formula (1) or a pharma- ceutically acceptable salt thereof: 【Chemistry 1】

9. 2. The pharmaceutical composition according to claim 1, wherein the cholinesterase inhibitor is donepezil represented by the following formula (2) or a pharma- ceutically acceptable salt thereof: 【Chemistry 2】

10. A food composition for preventing or improving neurodegenerative diseases, comprising a beta blocker and a cholinesterase inhibitor as active ingredients.

11. The food composition according to claim 10, characterized in that the food composition is a food for specified health uses.

12. Use of a beta blocker and a cholinesterase inhibitor for the manufacture of a pharmaceutical composition for the treatment of a neurodegenerative disease.

13. A method for treating a neurodegenerative disease, comprising administering to an individual in need thereof an effective amount of a composition comprising as active ingredients a beta blocker and a cholinesterase inhibitor.

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