Composition for preventing or treating alzheimer's disease comprising novel compound

Abstract

The present invention relates to the use of a novel compound for preventing, alleviating or treating Alzheimer's disease, and the novel compound exhibits an inhibitory effect on tau protein aggregation. In addition, it has been identified that a reduction in presenilin 1 is made by using the novel compounds for treatment. Therefore, the novel compound can be effectively used in the development of a therapeutic agent for Alzheimer's disease.

Description

Technical Field

[0001] This disclosure relates to compositions for the prevention or treatment of Alzheimer's disease, said compositions comprising novel compounds. Background Technology

[0002] Alzheimer's disease (AD) is a condition characterized by the progressive deterioration of memory, behavior, language, and spatiotemporal abilities. In the brains of Alzheimer's patients, plaque formation caused by β-amyloid protein has been observed, along with the accumulation of intracellular neurofibrillary tangles composed of paired helical filaments (PHFs) containing hyperphosphorylated tau. As a result, the entire brain tissue undergoes degeneration and atrophy, thus inducing the aforementioned symptoms.

[0003] In Alzheimer's disease, a deficiency of neurotransmitters such as acetylcholine is a key indicator, and restoring this deficiency has become one of the main treatment goals.

[0004] Parasympathomimetic drugs used for Alzheimer's disease can be classified into muscarinic agonists, nicotinic agonists, cholinesterase inhibitors (ChEIs), and drugs that indirectly regulate acetylcholine release. Among these, many ChEIs have been developed and are in use, including tacrine, physostigmine, donepezil, rivastigmine, and memantine.

[0005] Compared to first-generation drugs such as tacrine, second-generation drugs such as donepezil and rivastigmine have a longer duration of action, better stability, and higher blood-brain barrier (BBB) ​​permeability, resulting in higher concentrations within the central nervous system. While first-generation drugs non-selectively inhibit AChE, butyrylcholinesterase, and other peripheral cholinesterases, several newer drugs are known to have higher selectivity for AChE, thereby reducing peripheral side effects. These agents are known to exert their pharmacological effects by inhibiting the breakdown of acetylcholine (ACh), thus increasing intrasynaptic ACh concentrations.

[0006] However, conventional treatments for Alzheimer's disease are associated with serious side effects with long-term use. Therefore, there is a need to develop new drugs that exhibit equal or better pharmacological efficacy while reducing these adverse reactions. Summary of the Invention

[0007] Technical goals

[0008] Therefore, the purpose of this disclosure is to provide uses for the prevention, improvement or treatment of Alzheimer's disease, including novel compounds.

[0009] Technical solution

[0010] To achieve the above objectives, this disclosure provides pharmaceutical compositions for the prevention or treatment of Alzheimer's disease, said pharmaceutical compositions comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

[0011] [Chemical Formula 1]

[0012]

[0013] Wherein, in the chemical formula 1,

[0014] Is it a single or double bond?

[0015] n is an integer between 0 and 1.

[0016] X is CH or N, and

[0017] R 1 and R 2 They can be the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

[0018] In addition, this disclosure provides a health functional food composition for the prevention or improvement of Alzheimer's disease, the food composition comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0019] Beneficial effects

[0020] This disclosure relates to the use of a novel compound for the prevention, improvement, or treatment of Alzheimer's disease, wherein the novel compound (PRG-A) exhibits an inhibitory effect on tau protein aggregation. Furthermore, treatment with the PRG-A compound has been shown to cause a reduction in prosensory 1. Therefore, the PRG-A compound can be effectively used in the development of a therapeutic agent for Alzheimer's disease. Attached Figure Description

[0021] Figure 1 The effects of PRG-A compounds in preventing tau tangles are shown. A and B. Compounds PRG-A-01 to PRG-A-04 eliminated tau aggregation induced by overexpression of tau-WT and tau-MT. C. PRG-A compounds alleviated aberrant tubulin formation caused by overexpression of tau-MT.

[0022] Figure 2The effects of PRG-A compounds on the elimination of presenilin-1 (PS1) aggregation are shown. A and B. Compounds PRG-A-01 and PRG-A-02 reduce PS1-1 (PS1) aggregation induced by PS1 overexpression. C. PRG-A compounds inhibit PS1 aggregation in a dose-dependent manner.

[0023] Figure 3 The cytotoxic effects of the PRG-A compound were demonstrated. Detailed Implementation

[0024] This disclosure will be described in detail below.

[0025] This disclosure provides pharmaceutical compositions for the prevention or treatment of Alzheimer's disease, the pharmaceutical compositions comprising a compound represented by the following chemical formula 1, its hydrate or salt thereof.

[0026] [Chemical Formula 1]

[0027]

[0028] In the chemical formula 1,

[0029] Is it a single or double bond?

[0030] n is an integer between 0 and 1.

[0031] X is CH or N, and

[0032] R 1 and R 2 They can be the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

[0033] Preferably, in the compound, when When it is a single bond, n is 1, X is CH, and R is... 1 and R 2 They can be the same or different and each independently selected from (C1~C4)alkoxy, hydroxy or (C1~C4)alkylcarbonyloxy, but are not limited thereto.

[0034] Preferably, in the compound, when When it is a double bond, n is an integer from 0 to 1, X is CH or N, and R 1 and R 2 They can be the same or different and each independently selected from hydrogen, (C1~C4)alkoxy, hydroxyl, halogen, nitro or (C1~C4)alkylcarbonyloxy, but are not limited thereto.

[0035] Preferably, the compound can be a compound represented by the following chemical formula 2.

[0036] [Chemical Formula 2]

[0037]

[0038] In the chemical formula 2,

[0039] n is an integer between 0 and 1, and

[0040] R 1 'and R 2 'They can be the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

[0041] Compounds represented by chemical formula 1 or chemical formula 2 can be named PRG-A compounds.

[0042] More preferably, the compound may be selected from, but is not limited to, the group consisting of: (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl(E)-3-(4-hydroxy-3-methoxyphenyl)acrylate (PRG-A-01), (S,E)-7-((3-(4-hydroxy-3-methoxyphenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-one (PRG-A-02), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl3-(4-hydroxy-3-methoxy) (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3,4-dimethoxyphenyl)acrylate (SNU-C5), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl3-(3,4-dimethoxyphenyl)propionate (SNU-C7), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(pyridin-4-yl)acrylate (SNU-C9), (S)-8,8-dimethyl- 2-Oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromeno-7-yl(E)-3-(3-hydroxyphenyl)acrylate (SNU-C10), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromeno-7-yl(E)-3-(4-fluorophenyl)acrylate (PRG-A-03), (S,E)-7-((3-(4-fluorophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromeno-2-one (SNU-C13), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromeno-2-one (SNU-C13), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g] Tinno-7-yl(E)-3-(3-acetoxyphenyl)acrylate (SNU-C14), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinno-7-yl 3-(4-acetoxy-3-methoxyphenyl)propionate (SNU-C15), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinno-7-yl(E)-3-(4-acetoxy-3-methoxyphenyl)acrylate (SNU-C17), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinno-7-yl(E)-3-(3,4-Difluorophenyl)acrylate (SNU-C18) and (S,E)-7-((3-(3-methoxy-4-nitrophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromone-2-one (PRG-A-04).

[0043] More preferably, the pharmaceutical composition can inhibit the aggregation of tau protein and presenilin 1.

[0044] In addition to the active ingredient, the pharmaceutical compositions of this disclosure can also be prepared using pharmaceutically suitable and physiologically acceptable adjuvants, which may include solubilizers such as excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, flow aids, or flavoring agents. For administration, in addition to the active ingredient, the pharmaceutical compositions of this disclosure may also contain one or more pharmaceutically acceptable carriers to preferably formulate the pharmaceutical composition. In compositions formulated as liquid solutions, acceptable pharmaceutical carriers may be those that are sterile and biocompatible and used by mixing saline solutions, sterile water, Ringer's solution, buffered saline, albumin injection solutions, dextran solutions, maltodextrin solutions, glycerol, ethanol, and one or more mixtures of these components, and other conventional additives, such as antioxidants, buffers, and antibacterial agents, may be added as needed. Furthermore, diluents, dispersants, surfactants, binders, and lubricants may be added additionally to formulate injectable formulations (e.g., aqueous solutions, suspensions, and emulsions), pills, capsules, granules, or tablets.

[0045] The pharmaceutical compositions of this disclosure can be formulated as sustained-release dosage forms of the active compound, as well as granules, powders, coated tablets, tablets, capsules, suppositories, syrups, elixirs, suspensions, emulsions, drops, or injections. The pharmaceutical compositions of this disclosure can be administered conventionally via intravenous, intra-arterial, intra-peritoneal, intramuscular, intra-arterial, intra-sternal, percutaneous, intranasal, inhalation, topical, rectal, oral, intraocular, or intradermal routes. The effective amount of the active ingredient in the pharmaceutical compositions of this disclosure refers to the amount required for the prevention or treatment of a disease. Therefore, it can be adjusted according to various factors, including the type and severity of the disease, the type and content of the active ingredient and other components contained in the composition, the type of formulation, and the patient's age, weight, general health condition, sex, and diet, as well as the secretion rate of the composition, the time of administration, the route of administration, the treatment duration, and concurrent medications. Although not limited thereto, in the case of adults, for example, when administered once or several times a day, the compositions of this disclosure can be administered at doses from 0.01 ng / kg to 10 g / kg.

[0046] In addition, this disclosure provides a health functional food composition for the prevention or improvement of Alzheimer's disease, the food composition comprising a compound represented by the following chemical formula 1, its hydrate or salt thereof.

[0047] [Chemical Formula 1]

[0048]

[0049] In the chemical formula 1,

[0050] Is it a single or double bond?

[0051] n is an integer between 0 and 1.

[0052] X is CH or N, and

[0053] R 1 and R 2 They can be the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

[0054] Preferably, in the compound, when When it is a single bond, n is 1, X is CH, and R is... 1 and R 2 They can be the same or different and each independently selected from (C1~C4)alkoxy, hydroxy or (C1~C4)alkylcarbonyloxy, but are not limited thereto.

[0055] Preferably, in the compound, when When it is a double bond, n is an integer from 0 to 1, X is CH or N, and R 1 and R 2 They can be the same or different and each independently selected from hydrogen, (C1~C4)alkoxy, hydroxyl, halogen, nitro or (C1~C4)alkylcarbonyloxy, but are not limited thereto.

[0056] Preferably, the compound can be a compound represented by the following chemical formula 2.

[0057] [Chemical Formula 2]

[0058]

[0059] In the chemical formula 2,

[0060] n is an integer between 0 and 1, and

[0061] R 1 'and R 2'They can be the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

[0062] Compounds represented by chemical formula 1 or chemical formula 2 can be named PRG-A compounds.

[0063] More preferably, the compound may be selected from, but is not limited to, the group consisting of: (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl(E)-3-(4-hydroxy-3-methoxyphenyl)acrylate (PRG-A-01), (S,E)-7-((3-(4-hydroxy-3-methoxyphenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-one (PRG-A-02), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl3-(4-hydroxy-3-methoxy) (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3,4-dimethoxyphenyl)acrylate (SNU-C5), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl3-(3,4-dimethoxyphenyl)propionate (SNU-C7), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(pyridin-4-yl)acrylate (SNU-C9), (S)-8,8-dimethyl- 2-Oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromeno-7-yl(E)-3-(3-hydroxyphenyl)acrylate (SNU-C10), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromeno-7-yl(E)-3-(4-fluorophenyl)acrylate (PRG-A-03), (S,E)-7-((3-(4-fluorophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromeno-2-one (SNU-C13), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromeno-2-one (SNU-C13), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g] Tinno-7-yl(E)-3-(3-acetoxyphenyl)acrylate (SNU-C14), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinno-7-yl 3-(4-acetoxy-3-methoxyphenyl)propionate (SNU-C15), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinno-7-yl(E)-3-(4-acetoxy-3-methoxyphenyl)acrylate (SNU-C17), (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinno-7-yl(E)-3-(3,4-Difluorophenyl)acrylate (SNU-C18) and (S,E)-7-((3-(3-methoxy-4-nitrophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromone-2-one (PRG-A-04).

[0064] The health functional food compositions disclosed herein may further comprise one or more additives selected from the group consisting of: organic acids, phosphates, antioxidants, lactose casein, dextrin, glucose, sugars, and sorbitol. Organic acids may be, but are not limited to, citric acid, fumaric acid, adipic acid, lactic acid, or malic acid; phosphates may be, but are not limited to, sodium phosphate, potassium phosphate, acid pyrophosphate, or polyphosphate; and antioxidants may be, but are not limited to, natural antioxidants such as polyphenols, catechins, α-tocopherol, rosemary extract, licorice extract, chitosan, tannic acid, or phytic acid.

[0065] In another specific example of this disclosure, in addition to the active ingredients, the health functional food may also contain various nutrients, vitamins, minerals (electrolytes), flavoring agents (e.g., synthetic and natural flavoring agents), coloring agents and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, and carbonating agents used in carbonated beverages. Furthermore, a food composition according to one embodiment of this disclosure may contain fruit pulp used in the production of natural fruit juices, fruit juice beverages, and vegetable beverages.

[0066] According to one embodiment of this disclosure, the formulation of a health functional food may be in the form of a solid, powder, granules, tablet, capsule, liquid, or beverage, but is not limited thereto.

[0067] In addition, health functional foods can be used in the manufacture of foods, such as, but not limited to, sweets, sugar, ice cream products, dairy products, meat products, fish products, tofu or jelly, edible oils, noodles, tea, beverages, special nutritional foods, health products, condiments, frozen foods, ginseng products, pickled foods, dried foods, fruits, vegetables, dried fruits or vegetables, cut products, fruit juices, vegetable juices, their mixtures, potato chips, noodles, processed livestock and poultry foods, processed seafood foods, processed dairy foods, fermented dairy foods, legume foods, cereal foods, microbially fermented foods, pastries and baked goods, seasonings, processed meat products, acidic beverages, licorice and herbs.

[0068] Example

[0069] In the following description, embodiments will be presented in detail to aid in understanding this disclosure. However, the following embodiments are intended to illustrate the content of this disclosure only, and the scope of this disclosure is not limited to the following embodiments. The embodiments of this disclosure are provided to describe this disclosure more completely to those skilled in the art.

[0070] <Experimental Example>

[0071] The following experimental examples are intended to provide experimental examples that are typically applied to each embodiment of this disclosure.

[0072] 1. Compound

[0073] The chemical structures and NMR data of the compounds used in this disclosure are as follows.

[0074] (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(4-hydroxy-3-methoxyphenyl)acrylate (PRG-A-01)

[0075]

[0076] 1 1H NMR (acetone-d6, 400MHz)

[0077] δ ppm 7.843(d, J=9.6Hz, 1H), 7.616(d, J=16.0Hz, 1H), 7.424(s, 1H), 7.357(s, 1H), 7.123(dd, J=2.0, 8.0Hz, 1H), 6.851(d, J=8.4Hz, 1H), 6.740(s,1H), 6.387(d, J=15.6Hz, 1H), 6.198(d, J=9.6Hz, 1H), 5.221(t, J=4.6Hz, 1H),3.895(s, 3H), 3.321(dd, J=4.6, 17.2Hz, 1H), 2.963(dd, J=4.4, 17.6Hz, 1H),1.421(s, 3H), 1.413(s, 3H); MS(m / z) 423 (M+H) + .

[0078] (S,E)-7-((3-(4-hydroxy-3-methoxyphenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-one (PRG-A-02)

[0079]

[0080] 1 H NMR: EW15731-164-P1D4 (400 MHz, CDCl3).

[0081] δ 7.58 (d, J = 12 Hz, 1H), 7.16 (s, 1H), 6.90 - 6.86 (m, 3H), 6.78(s, 1H), 6.51 (d, J =16 Hz, 1H), 6.22 (d, J = 8.0 Hz, 1H), 6.14 - 6.10 (m,1H), 5.65 (s, 1H), 4.32 - 4.30 (m, 1H), 4.22- 4.20 (m, 1H), 3.91 (s, 3H), 3.61 - 3.58 (m, 1H), 3.11 - 3.05 (m, 1H), 3.89 - 3.87 (m, 1H), 1.43 (s, 3H), 1.36 (s, 3H).

[0082] (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(4-fluorophenyl)acrylate (PRG-A-03)

[0083]

[0084] 1 H NMR (CD2Cl2, 400MHz): δ ppm 7.66 - 7.55 (m, 2H), 7.55 - 7.46 (m,2H), 7.18 (s, 1H), 7.11 - 7.01 (m, 2H), 6.76 (s, 1H), 6.35 (d, J = 16.0 Hz,1H), 6.16 (d, J = 9.5 Hz, 1H), 5.17 (app.t, J = 4.7 Hz, 1H), 3.23 (ddd, J =17.3, 4.8, 1.2 Hz, 1H), 2.92 (dd, J = 17.3, 4.7 Hz, 1H), 1.40 (s, 3H), 1.36 (s, 3H).

[0085] (S,E)-7-((3-(3-methoxy-4-nitrophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-2-one (PRG-A-04)

[0086]

[0087] 1 H NMR (CDCl3, 400MHz): δ ppm 7.85 (d, J = 8.4 Hz, 1H), 7.57 (d, J =9.5 Hz, 1H), 7.16 (s, 1H), 7.05 - 6.97 (m, 2H), 6.78 (s, 1H), 6.60 4.24 (ddd, J = 13.4, 5.5, 1.6Hz, 1H), 3.97 (s, 3H), 3.59 (dd, J = 7.1, 4.9 Hz, 1H), 3.11 (dd, J = 16.7,4.9 Hz, 1H), 2.87 (dd, J = 16.7, 7.2 Hz, 1H), 1.43 (s, 3H), 1.38 (s, 3H).

[0088] (S,E)-7-((3-(4-fluorophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-one (SNU-C13)

[0089]

[0090] 1H NMR (CDCl3, 400MHz): δ ppm 7.57 (d, J = 9.5 Hz, 1H), 7.38 - 7.29(m, 2H), 7.16 (s, 1H), 7.06 - 6.93 (m, 2H), 6.77 (s, 1H), 6.55 (d, J = 15.8Hz, 1H), 6.25 - 6.13 (m, 2H), 4.33 (ddd, J = 12.8, 5.8, 1.5 Hz, 1H), 4.19 (ddd, J = 12.8, 6.3, 1.4 Hz, 1H), 3.58 (dd, J = 7.4, 5.0 Hz, 1H), 3.08 (dd, J= 16.4, 5.0 Hz, 1H), 2.85 (ddd, J = 16.4, 7.4, 1.1 Hz, 1H), 1.42 (s, 3H), 1.35 (s, 3H).

[0091] (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(4-acetoxy-3-methoxyphenyl)acrylate (SNU-C17)

[0092]

[0093] 1 H NMR (CD2Cl2, 600MHz): δ ppm 7.67 - 7.56 (m, 2H), 7.21 (s, 1H), 7.14- 7.09 (m, 2H), 7.03 (d, J = 8.0 Hz, 1H), 6.78 (s, 1H), 6.41 (d, J = 16.0 Hz, 1H), 6.19 (d, J = 9.5 Hz, 1H), 5.20 (app.t, J = 4.7 Hz, 1H), 3.82 (s, 3H), 3.26 (ddd, J = 17.3, 4.8, 1.2 Hz, 1H), 2.95 (dd, J = 17.3, 4.5 Hz, 1H), 2.27(s, 3H), 1.43 (s, 3H), 1.38 (s, 3H).

[0094] (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3,4-difluorophenyl)acrylate (SNU-C18)

[0095]

[0096] 1 H NMR (CDCl3, 400MHz): δ ppm 7.62 - 7.53 (m, 2H), 7.32 (ddd, J =11.1, 7.6, 2.2 Hz, 1H), 7.24 - 7.11 (m, 3H), 6.83 (s, 1H), 6.33 (d, J = 16.0,1H), 6.24 (d, J = 9.4 Hz, 1H), 5.19 (app.t, J = 4.7 Hz, 1H), 3.25 (ddd, J =17.4, 4.8, 1.1 Hz, 1H), 2.93 (dd, J = 17.4, 4.5 Hz, 1H), 1.43 (s, 3H), 1.39 (s, 3H).

[0097] (7S)-(+)-3-(2-furanyl)-acrylic acid, 8,8-dimethyl-2-oxo-6,7-dihydro-2H,8H-pyrano[3,2-g]chromene-7-yl-ester (SLC-B050)

[0098]

[0099] 1 H NMR (400 MHz, CDCl3): δ H 7.64(1H, s, H-6'), 7.58(1H, d, J = 9.6Hz, H-4), 7.56(1H, d, J = 16.0Hz, H-3'), 7.41(1H, d, J = 1.6Hz, H-7'), 7.17(1H,s, H-5), 6.82(1H, s, H-10), 6.55(1H, d, J = 1.6Hz, H-8'), 6.23(1H, d, J =9.6Hz, H-3), 6.13(1H, d, J = 16.0Hz, H-2'), 5.17(1H, t, J = 4.4Hz, H-7), 3.23(1H, dd, J = 4.4, 17.6Hz, H-6a), 2.92(1H, dd, J = 4.4, 17.6Hz, H-6b), 1.42(3H, s, CH3-8), 1.38(3H, s, CH3-8).

[0100] 2. Cell Culture and Reagents

[0101] HEK293 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained in DMEM medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin under a humid atmosphere of 37°C and 5% CO2. SK-N-SH cells were purchased from the Korean Cell Bank (KCLB, Seoul, South Korea) and maintained in MEM medium supplemented with 10% fetal bovine serum, 1% antibiotics, 25 mM HEPES, and 300 mg / L L-glutamine. Human fibroblasts (9-year-old female) were obtained from the Coriell Cell Bank (New Jersey, USA) and maintained in EMEM medium supplemented with 15% FBS, 2 mM glutamine, and 26 mM HEPES, but without antibiotics.

[0102] 3. Western blot analysis

[0103] For SDS-PAGE, proteins were extracted from cells using RIPA buffer (50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.1% SDS, and 10% sodium deoxycholate). Samples were separated by SDS-PAGE and transferred to PVDF membranes. The blotted membranes were blocked with 3% skim milk containing TBST buffer for 1 h and incubated with specific antibodies. The reacted antibodies were detected by ECL and X-ray film exposure. HA (sc-7392) and GFP (green fluorescent protein; sc-8036) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Actin antibody (66009-1-lg) and α-tubulin antibody (66240-1-lg) were obtained from Proteintech (Rosemont, IL, USA). Anti-TH antibody (NB300-109) was purchased from Novus Biologicals (CO, USA). HRP-conjugated goat anti-mouse antibody, goat anti-rabbit antibody, and mouse anti-goat antibody (Pierce, Thermo Fisher Scientific, Inc., Rockford, IL, USA) were used as secondary antibodies. After incubation, cells were harvested using TNN buffer (50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.3% NP-40) and centrifuged at 14,000 rpm for 30 min to separate the precipitate (insoluble) and supernatant (soluble). “Input” represents whole-cell lysate (WCL) harvested using RIPA buffer. Actin was used as a loading control.

[0104] 4. Immunofluorescence staining

[0105] Cells on coverslips were washed with PBS, fixed with 4% paraformaldehyde (PFA) at room temperature for 30 min, and then permeabilized in 0.1% Triton X-100 / PBS for 10 min. After treating cells with blocking solution (3% anti-human antibody diluted 1:500 in PBS) for 1 hour, cells were incubated with HA (1:500 diluted) at 4°C overnight. Finally, cells were incubated with FITC- and rhodamine-conjugated secondary antibodies at 4°C for 6 hours. Cell nuclei were stained with 4,6-diamidinyl-2-phenylindole (DAPI), and endoplasmic reticulum (ER) was stained with ER-Tracker Red dye for 10 min. Cells were washed three times with PBS, and then mounted with mounting solution (H-5501; Vector Laboratories, Burlingame, CA, USA) for analysis using a fluorescence microscope (Zeiss).

[0106] 5. Transfection of the vector

[0107] GFP-Tau (WT; #46904, P301L; #46908) plasmids were purchased from Addgene (Cambridge, MA, USA). Transfection was performed using Jet-PEI reagent (JetPEI; Polyplus transfection, New York, NY, USA) according to the manufacturer's protocol. In brief, the vector was mixed with JetPEI reagent in 150 mM NaCl buffer and incubated for 15 minutes. The mixture was then added to cells in serum-free medium and incubated for 4 hours. After incubation, the cells were replaced with medium supplemented with 10% FBS.

[0108] 6. Measurement of cell viability

[0109] To study cell viability, cells were treated with a chemical (PRG-A compound) for 48 h. Then, the cells were incubated with 0.5 mg / mL 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution (475989; Merck, Darmstadt, Germany) at 37 °C for 4 h. After removing excess solution and washing with PBS, the precipitate was dissolved in 200 μL DMSO and quantified by measuring absorbance at 540 nm.

[0110] <Example 1>

[0111] To investigate the effect of PRG-A compounds on tau aggregation (tau tangles), the inventors transfected HEK293 cells with indicator vectors (Tau-WT, Tau-P301L) and treated the cells with the selected compound (5 μM) for 24 hours. Cells were harvested using TNN buffer and centrifuged at 14,000 rpm for 30 minutes to separate the precipitate (insoluble) and the supernatant (soluble). Figure 1 A and Figure 1 B). After transfection, the aggregated mutant tau (P301L) was recovered as an insoluble fraction. Figure 1 A). However, treatment with PRG-A-01 and PRG-A-02 significantly reduced the insoluble mutant tau fraction (A). Figure 1 A). Similar effects were observed when treated with PRG-A-04 ( Figure 1 B). In addition, cells were transfected with the Tau-P301L vector for 24 hours. After incubation, cells were treated again with PRG-A compound (5 μM) for 24 hours. Cells were fixed with 4% PFA and immunofluorescence was performed using α-tubulin antibody (white arrow, high tau intensity). Figure 1 C). As a result, abnormal tubulin arrangement (yellow arrow) and mutant tau aggregates (white arrow) were normalized or dissolved by the PRG-A compound ( Figure 1 C). The compound Chem-001 (imperatorin) showed only a weak effect in inhibiting tau aggregation.

[0112] <Example 2>

[0113] It is also known that mutations in prosensory 1 (PS1) are associated with Alzheimer's disease (AD). Therefore, to examine the effect of PRG-A compounds on PS1 expression and localization, cells were transfected with PS1-HA or PS1-GFP vectors for 24 hours. After incubation, cells were treated with PRG-A compound (5 μM) for 24 hours. Cells were fixed with 4% PFA and stained with HA antibody (green) and ER markers (red) to monitor PS1 inclusion bodies (white arrows, high PS1 intensity). Figure 2 A and Figure 2 B). Ectopic PS1 expression forms aggregates in the cytoplasm ( Figure 2 A and Figure 2 B). In contrast, the PRG-A compound was able to dissolve PS1 aggregates in IF staining ( Figure 2 A and Figure 2B). To confirm the PS1 aggregation inhibitory effect of the PRG-A compounds, SK-N-SH cells were transfected with an indicator vector (PS1-HA) and treated with selected compounds (PRG-A-01 and PRG-A-02) for 24 hours. SDS-PAGE was performed using HA antibody. Figure 2 C). In fact, oligomeric PS1 is reduced by treatment with PRG-A compounds ( Figure 2 C). In summary, these results indicate that PRG-A compounds may also be used for the prevention or treatment of Alzheimer's disease.

[0114] <Example 3>

[0115] Finally, to determine the toxicity of the PRG-A compounds, normal human fibroblasts and neuroblastoma cells (SK-N-SH) were cultured with each compound for 48 hours, and cell viability was measured by the MTT assay. The effects on the survival of neurons and normal fibroblasts were identified. Figure 3 Compounds PRG-A-01, PRG-A-02, and PRG-A-04 did not exhibit cytotoxic effects in normal human fibroblasts. Compound PRG-A-03 showed low effects on cell viability at high doses (50 μM and 100 μM). In summary, the novel compounds did not show toxicity in normal cells up to 50 μM, therefore, the side effects of these novel compounds are not expected to be severe.

[0116] In summary, this novel compound can be used to treat Alzheimer's disease (AD) without serious side effects.

[0117] Although specific portions of this disclosure have been described in detail above, it will be apparent to those skilled in the art that this detailed description is merely a preferred exemplary embodiment, and the scope of this disclosure is not limited thereto. In other words, the essential scope of this disclosure is defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition for the prevention or treatment of Alzheimer's disease, said pharmaceutical composition comprising a compound represented by the following chemical formula 1, its hydrate, or a salt thereof: [Chemical Formula 1] in, In the chemical formula 1, Is it a single or double bond? n is an integer between 0 and 1. X is CH or N, and R 1 and R 2 They may be the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

2. The pharmaceutical composition according to claim 1, wherein, In the compound, when When it is a single bond, n is 1, X is CH, and R is... 1 and R 2 They are the same or different and are each independently selected from (C1~C4)alkoxy, hydroxy or (C1~C4)alkylcarbonyloxy.

3. The pharmaceutical composition according to claim 1, wherein, In the compound, when When it is a double bond, n is an integer from 0 to 1, X is CH or N, and R 1 and R 2 They are the same or different and are each independently selected from hydrogen, (C1~C4)alkoxy, hydroxyl, halogen, nitro or (C1~C4)alkylcarbonyloxy.

4. The pharmaceutical composition according to claim 1, wherein, The compounds are selected from the group consisting of: (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl(E)-3-(4-hydroxy-3-methoxyphenyl)acrylate, (S,E)-7-((3-(4-hydroxy-3-methoxyphenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-one, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl 3-(4-hydroxy-3-methoxyphenyl)propionate, (S)-8,8-dimethyl-2-oxo 7,8-Dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3,4-dimethoxyphenyl)acrylate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl 3-(3,4-dimethoxyphenyl)propionate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(pyridin-4-yl)acrylate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3-hydroxyphenyl) Acrylates, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(4-fluorophenyl)acrylates, (S,E)-7-((3-(4-fluorophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-2-one, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3-acetoxyphenyl)acrylates, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3-acetoxyphenyl)acrylates, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g] Tinmen-7-yl 3-(4-acetoxy-3-methoxyphenyl)propionate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinmen-7-yl(E)-3-(4-acetoxy-3-methoxyphenyl)acrylate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinmen-7-yl(E)-3-(3,4-difluorophenyl)acrylate, and (S,E)-7-((3-(3-methoxy-4-nitrophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]tinmen-2-one.

5. The pharmaceutical composition according to claim 1, wherein, The compound is represented by the following chemical formula 2: [Chemical Formula 2] In the chemical formula 2, n is an integer between 0 and 1, and R 1 'and R 2 'They are the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

6. The pharmaceutical composition according to claim 1, wherein, The pharmaceutical composition inhibits the aggregation of tau protein and presenilin 1.

7. A health functional food composition for preventing or improving Alzheimer's disease, said health functional food composition comprising a compound represented by the following chemical formula 1, its hydrate, or a salt thereof: [Chemical Formula 1] in, In the chemical formula 1, Is it a single or double bond? n is an integer between 0 and 1. X is CH or N, and R 1 and R 2 They may be the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

8. The health functional food composition according to claim 7, wherein, In the compound, when When it is a single bond, n is 1, X is CH, and R is... 1 and R 2 They are the same or different and are each independently selected from (C1~C4)alkoxy, hydroxy or (C1~C4)alkylcarbonyloxy.

9. The health functional food composition according to claim 7, wherein, In the compound, when When it is a double bond, n is an integer from 0 to 1, X is CH or N, and R 1 and R 2 They are the same or different and are each independently selected from hydrogen, (C1~C4)alkoxy, hydroxyl, halogen, nitro or (C1~C4)alkylcarbonyloxy.

10. The health functional food composition according to claim 7, wherein, The compounds are selected from the group consisting of: (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl(E)-3-(4-hydroxy-3-methoxyphenyl)acrylate, (S,E)-7-((3-(4-hydroxy-3-methoxyphenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-one, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl 3-(4-hydroxy-3-methoxyphenyl)propionate, (S)-8,8-dimethyl-2-oxo 7,8-Dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3,4-dimethoxyphenyl)acrylate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl 3-(3,4-dimethoxyphenyl)propionate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(pyridin-4-yl)acrylate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3-hydroxyphenyl) Acrylates, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(4-fluorophenyl)acrylates, (S,E)-7-((3-(4-fluorophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-2-one, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3-acetoxyphenyl)acrylates, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromene-7-yl(E)-3-(3-acetoxyphenyl)acrylates, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g] Tinmen-7-yl 3-(4-acetoxy-3-methoxyphenyl)propionate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinmen-7-yl(E)-3-(4-acetoxy-3-methoxyphenyl)acrylate, (S)-8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]tinmen-7-yl(E)-3-(3,4-difluorophenyl)acrylate, and (S,E)-7-((3-(3-methoxy-4-nitrophenyl)allyl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]tinmen-2-one.

11. The health functional food composition according to claim 7, wherein, The compound is represented by the following chemical formula 2: [Chemical Formula 2] In the chemical formula 2, n is an integer between 0 and 1, and R 1 'and R 2 'They are the same or different, and each is independently selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, hydroxyl, halogen, nitro, cyano or (C1~C4)alkylcarbonyloxy.

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