KR20230157255A - Composition for preventing, ameliorating or treating Alzheimer's disease comprising TKIM as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating Alzheimer's disease containing TKIM as an active ingredient. In an animal model in which Alzheimer's disease is induced by administering amyloid beta, cognitive, memory and spatial learning abilities are improved upon administration of TKIM. Since the recovery effect is excellent, the composition containing TKIM of the present invention as an active ingredient can be usefully used as a treatment for Alzheimer's disease or as a health functional food for improving Alzheimer's disease.

Description

Composition for preventing, ameliorating or treating Alzheimer's disease comprising TKIM as effective component}

The present invention relates to TKIM [4-chloro-N'-(2-(quinolin-2-ylthio)acetyl)benzohydrazide; CAS 326921-25-9] as an active ingredient and relates to a composition for preventing, improving or treating degenerative brain diseases.

Alzheimer's disease is the most common degenerative brain disease that causes dementia, and was first reported in 1907 by German psychiatrist Dr. Alois Alzheimer. Alzheimer's disease is characterized by a very slow onset and gradual progression. In the beginning, problems mainly occur with memory for recent events, but as the disease progresses, abnormalities in other cognitive functions such as language function and judgment occur, and eventually, all daily life functions are lost.

As Alzheimer's disease progresses, it is often accompanied by not only cognitive decline but also mental and behavioral symptoms such as personality changes, agitated behavior, depression, delusions, hallucinations, increased aggression, and sleep disorders. In the later stages, neurological disorders such as stiffness and gait abnormalities occur. Or, physical complications such as incontinence of urine and feces, infection, and bedsores may occur.

When examining the brain tissue of a patient with Alzheimer's disease under a microscope, characteristic lesions such as neuritic plaques and neurofibrillary tangles are observed, and when observed with the naked eye, overall brain atrophy is seen due to neuronal loss. . In the early stages of the disease, these brain pathology findings are mainly limited to the hippocampus and entorhinal cortex, which are major brain regions responsible for memory, but gradually spread throughout the brain through the parietal lobe and frontal lobe.

As the brain pathology invades the area, memory decline mainly appears in the beginning, but as the disease progresses, clinical symptoms become more diverse and more severe as the disease progresses.

The exact pathogenesis and cause of Alzheimer's disease are not exactly known. Currently, it is known that the core mechanism of the disease is that a small protein called beta-amyloid is excessively produced and deposited in the brain, which has a harmful effect on brain cells. In addition, tau protein (Tau protein), which plays an important role in maintaining the skeleton of brain cells, Hyperphosphorylation of tau protein, inflammatory response, and oxidative damage also appear to contribute to brain cell damage and affect the development of the disease. Neurogenic plaques (or senile plaques), a representative brain pathology, are associated with deposition of beta-amyloid protein, and neurofibrillary tangles are associated with tau protein hyperphosphorylation.

Meanwhile, Korean Patent No. 1723514 discloses a pharmaceutical composition for preventing or treating Alzheimer's disease and degenerative brain diseases containing a SNO-OGT (S-nitrosylation of O-linked N -acetylglucosaminyltransferase) inhibitor, and Korean Patent No. 0890678 Although malonamide derivatives as gamma-secretase inhibitors for the treatment of Alzheimer's disease are disclosed, there is no disclosure of a composition for the prevention, improvement or treatment of Alzheimer's disease containing the TKIM of the present invention as an active ingredient.

The present invention was developed in response to the above-mentioned needs, and provides a composition for preventing, improving or treating Alzheimer's disease containing TKIM as an active ingredient, and applying the TKIM to an animal model in which Alzheimer's disease is induced by administering amyloid beta. The present invention was completed by confirming that cognition, memory, and spatial learning ability were restored upon administration.

In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention or treatment of Alzheimer's disease containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving Alzheimer's disease, containing the compound represented by Formula 1 or a foodologically acceptable salt thereof as an active ingredient.

Additionally, the present invention provides a veterinary composition for the prevention or treatment of Alzheimer's disease containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

Additionally, the present invention provides a feed additive for preventing or improving Alzheimer's disease containing the compound represented by Formula 1 or a foodologically acceptable salt thereof as an active ingredient.

The present invention relates to a composition for preventing, improving or treating Alzheimer's disease containing TKIM as an active ingredient, when TKIM is administered to an animal model in which Alzheimer's disease is induced by administering amyloid beta, the Y-maze test and the water maze test are performed. It has the effect of restoring the cognitive, memory, and spatial learning abilities that have been deteriorated due to Alzheimer's disease to normal levels by restoring the results to normal levels.

Figure 1 shows the results of a Y-maze experiment confirming the cognitive and memory improvement effect of TKIM administration in an animal model in which Alzheimer's disease was induced by amyloid beta administration. The number of crossings (alternation) was calculated as one crossing when the mouse sequentially passed through three passages in succession. * indicates a statistically significant decrease in the number of crossovers in the amyloid beta (Aβ _1-42 ) administration group compared to the normal control group (Saline), p<0.05. # means that the number of crossovers in the amyloid beta (Aβ _1-42 ) and TKIM administered group was statistically significantly increased compared to the amyloid beta (Aβ _1-42 ) alone administered group, p<0.05.
Figure 2 shows the results of a water maze test (Morris water maze) confirming long-term memory and spatial learning ability by TKIM administration in an animal model in which Alzheimer's disease was induced by administering amyloid beta. Training was completed, and the time to recognize and reach the escape zone (Latency to target) was measured on the 5th day. * means that the arrival time of the amyloid beta (Aβ _1-42 ) administered group was statistically significantly increased compared to the normal control group (Saline), p<0.05. # means that the arrival time of the amyloid beta (Aβ _1-42 ) and TKIM administered group was statistically significantly reduced compared to the amyloid beta (Aβ _1-42 ) alone administered group, p<0.05.

In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention or treatment of Alzheimer's disease containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

The Alzheimer's disease may be Alzheimer's mild cognitive impairment or Alzheimer's dementia, but is not limited thereto.

In the present invention, 'pharmaceutically acceptable salt' refers to any organic or inorganic addition salt that is relatively non-toxic to patients and does not cause side effects that reduce the beneficial efficacy of the compound, such as free acid, inorganic acid, Organic acids or non-toxic salts can be used. Preferred inorganic acids include hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, or tartaric acid. Preferred organic acids include methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, and maleic acid. (maleic acid), succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, glue Gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, or hydroiodic acid can be used. .

Acid addition salts such as the above inorganic acids or organic acids can be prepared by conventional methods, such as dissolving the compound in an excess of aqueous acid and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. there is. Equimolar amounts of the compound and acid or alcohol in water can be heated, and the mixture can then be evaporated to dryness, or the precipitated salt can be suction filtered.

The non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, and fluoride. , acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, Fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methyl benzoate, dinitro benzoate, hydroxybenzoate, methoxybenzoate phthalate , terephthalate, benzenesulfonate, toluene sulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, beta-hydroxybutyrate, glycolate, malate, Tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate or mandelate can be preferably used.

In the present invention, the term 'prevention' refers to any action that inhibits or delays the onset of Alzheimer's disease by administering the pharmaceutical composition according to the present invention, and 'treatment' refers to individuals suspected of or developing Alzheimer's disease by administering the pharmaceutical composition. It refers to any action that improves or changes the symptoms in a beneficial way.

In addition to the above active ingredients, it may further include a carrier, excipient, or diluent. The pharmaceutical composition of the present invention can be administered orally or parenterally, and when administered parenterally, it is preferable to select the method of external application to the skin or intraperitoneal, rectal, intravenous, intramuscular or subcutaneous injection, but is not limited thereto.

The pharmaceutical composition of the present invention may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain one or more compounds and at least one excipient, such as starch, calcium carbonate, sucrose, or lactose ( It is prepared by mixing lactose, gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspension solvents may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, Tween 61, cacao, laurel, glycerol, gelatin, etc. can be used.

The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, “pharmaceutically effective amount” means an amount sufficient to treat the disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, and activity of the patient's disease. , can be determined based on factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other factors well known in the field of medicine. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

The dosage of the composition of the present invention can vary depending on the patient's weight, age, gender, health condition, diet, administration time, administration method, excretion rate, and severity of the disease.

In addition, the present invention provides a health functional food composition for preventing or improving Alzheimer's disease, containing the compound represented by Formula 1 or a foodologically acceptable salt thereof as an active ingredient.

The health functional food composition can be manufactured as any one selected from pills, tablets, capsules, powders, powders, granules, candies, syrups and beverages, or by adding it as an ingredient in food, using a conventional method. It can be manufactured appropriately.

Examples of foods to which the active ingredient of the present invention can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, It may be in any form selected from tea, drink, alcoholic beverage, and vitamin complex, and includes all health functional foods in the conventional sense.

The health functional foods include various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, It may contain pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages. Additionally, it may contain pulp for the production of natural fruit juice and vegetable drinks. These ingredients can be used independently or in combination.

The health functional food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients. The natural carbohydrates include 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. As a sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used.

Additionally, the present invention provides a veterinary composition for the prevention or treatment of Alzheimer's disease containing the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

The veterinary composition of the present invention may further include appropriate excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostearate. , polysorbate 60, methylparaben, propylparaben, and mineral oil.

The veterinary composition according to the present invention may further include fillers, anti-aggregants, lubricants, wetting agents, spices, emulsifiers, preservatives, etc. The veterinary composition according to the present invention provides rapid and sustained release of the active ingredient after administration to an animal. or may be formulated using methods well known in the art to provide sustained release, the dosage form being powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, It may be in the form of a suppository, sterile injectable solution, or sterile topical medication.

The effective amount of the veterinary composition according to the present invention can be appropriately selected depending on the individual animal. Severity of the disease or condition, sensitivity to the active ingredient of the present invention depending on the individual's age, weight, health condition or gender, administration route, administration period, factors including other compositions mixed or used simultaneously with the composition, and other physiological or It can be determined based on factors well known in the veterinary field.

Additionally, the present invention provides a feed additive for preventing or improving Alzheimer's disease containing the compound represented by Formula 1 or a foodologically acceptable salt thereof as an active ingredient.

The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. In the present invention, the term 'feed' may mean any natural or artificial diet, meal, etc., or a component of the meal, for or suitable for eating, ingestion, and digestion by animals. The type of feed is not particularly limited, and feed commonly used in the art can be used. Non-limiting examples of the feed include plant feeds such as grains, roots and fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, cucurbits or grain by-products; Examples include animal feeds such as proteins, inorganic substances, fats and oils, minerals, oils and fats, single-cell proteins, zooplanktons or food. These may be used alone or in combination of two or more types.

Hereinafter, the present invention will be described in more detail using examples. These examples are only for illustrating the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited thereto.

Materials and Methods

1. Amyloid beta (Aβ) _1-42 ) Production of administration model

C57BL6 male mice aged 12 weeks were used as a wild type (WT) animal model.

The experimental animals had an adaptation period for one week before the experiment, and any abnormal behavior was observed. Animals with abnormal behavior were excluded from the experiment. During the experiment period, the experimental animals were allowed to drink water and feed freely under conditions of 12 hours of sunlight and 12 hours of shading in a temperature and humidity controlled breeding facility. Animal experiments were conducted in accordance with the guidelines of the Gyeongsang National University Animal Ethics Review Committee (GNU-200702-M0041, GNU-201021-M0079).

To create an amyloid beta administration model, the animal model was anesthetized by intramuscular injection of avertin and then placed in a stereotaxic brain surgery frame. For each animal model, 5 μl (100 μM) of amyloid beta (Aβ _1-42 ) oligomer was injected into the CA1 region of the hippocampus at a depth of 2.2 mm from the crown using a 10 μl Hamilton microsyringe equipped with a 26G needle to inject amyloid beta. (Aβ _1-42 ) administered Alzheimer's animal model was created.

2. Grouping and drug treatment of experimental mice

24 hours after amyloid beta was administered, the experimental mice were divided into three groups. They were classified into a normal control group administered phosphate buffer solution (PBS) instead of amyloid beta, an amyloid beta administered group, and an amyloid beta + 30μM TKIM administered group.

TKIM (CAS 326921-25-9) with a purity of over 98% was prepared by dissolving in 5% DMSO, and administered orally at a dosage of 11.2 mg/kg/day once a day for 1 week starting the day before amyloid beta administration. did.

The group that did not administer TKIM was orally administered the same amount of DMSO (0.05%). During the behavioral analysis experiment, the Y-maze test was measured 48 hours after amyloid beta administration. The water maze test was conducted from the third day of amyloid beta administration, and the final results were analyzed on the end of drug administration.

3. Behavior analysis

1) Measurement of spatial perception ability through Y-maze test

Spatial perception ability was measured by performing the Y-maze test. The movement of the animal model is expressed by the number of crossings (alternation), and the number of crossings refers to one crossing when the animal model sequentially passes through three passages in succession. Each animal model started by looking at the wall at the end of one maze and went to the other maze. If it successfully crossed, it was scored by giving 1 point. Scoring is done by dividing each score by the highest possible score (the number of passes through each arm minus 2) and multiplying it by 100. The criterion for passing the maze was when the animal model passed half of each passage.

2) Water maze test (Morris water maze)

The water maze test (Morris water maze) was performed to evaluate long-term memory and spatial learning ability. The temperature of the water used in the experiment was maintained at 22±2°C. The water maze circular pool (90 cm in diameter, 30 cm in height) was divided into four quadrants: N, S, E, and W. The water in the water maze was diluted with skim milk, and an escape zone 1 cm below the water surface was installed on one side of the quadrant. The experimental animals were made to swim and were trained to find the platform zone repeatedly in different positions once a day for 4 days. The movements of the experimental animals were recorded using a SMART video tracking system (Ethovision ^® XT software, Noldus Information Technology Inc. Leesburg, VA, USA). On the fifth day of the experiment, the time to recognize and reach the escape zone within 60 seconds was recorded.

4. Statistical analysis

Significance between groups depends on whether the normality assumption is satisfied, using non-parametric test methods such as Kruskal-Wallis analysis, Mann-Whitney analysis, and one-way analysis of variance (ANOVAOneWay)/Bonferroni. It was tested using (OriginPro2020, OriginLab Corp. Northampton, MA, USA). Data were expressed as mean ± standard deviation (mean ± SD), and the level of statistical significance was set at a p value of less than 0.05 (p < 0.05).

Example 1. Measurement of spatial perception ability through Y-maze test

After administering TKIM to an animal model in which Alzheimer's disease was induced by administering amyloid beta, a Y-maze experiment was performed. As a result, as shown in Figure 1, the number of crossings reduced by administration of amyloid beta was increased upon administration of TKIM, It was confirmed that cognition and memory were improved by TKIM administration.

Example 2. Water maze test (Morris water maze)

After administering TKIM to an animal model in which Alzheimer's disease was induced by administering amyloid beta, a water maze test was performed. As a result, as shown in Figure 2, the group in which Alzheimer's disease was induced by administering amyloid beta (Aβ _1-42 ), the time to recognize and reach the escape zone increases compared to the normal control group (Saline), but in the group administered amyloid beta and TKIM, the time to recognize and reach the escape zone recovered to the level of the normal control group, and TKIM It was confirmed that long-term memory and spatial learning ability were improved by administration.

Claims (8)

A pharmaceutical composition for the prevention or treatment of Alzheimer's disease containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]
The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, wherein the Alzheimer's disease is mild cognitive impairment of Alzheimer's disease or dementia of Alzheimer's disease. The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, further comprising a carrier, excipient, or diluent in addition to the active ingredient. The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, wherein the composition is prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols. A health functional food composition for preventing or improving Alzheimer's disease, containing a compound represented by the following formula (1) or a foodologically acceptable salt thereof as an active ingredient.
[Formula 1]
The method of claim 5, wherein the composition is prepared in any one formulation selected from pills, tablets, capsules, powders, powders, granules, candies, syrups and beverages. Health functional food composition for improvement. A veterinary composition for the prevention or treatment of Alzheimer's disease containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]
A feed additive for preventing or improving Alzheimer's disease containing a compound represented by the following formula (1) or a foodologically acceptable salt thereof as an active ingredient.
[Formula 1]