WO2016167385A1

WO2016167385A1 - Composition-ii for preventing or treating alzheimer's disease, comprising lycopodiella cernua extract or compound isolated therefrom

Info

Publication number: WO2016167385A1
Application number: PCT/KR2015/003790
Authority: WO
Inventors: 민병선; 최재수
Original assignee: 대구가톨릭대학교산학협력단
Priority date: 2015-04-14
Filing date: 2015-04-15
Publication date: 2016-10-20
Also published as: KR101818084B1; KR20160122928A

Abstract

The present invention relates to a composition for preventing or treating Alzheimer's disease comprising lycopodiella cernua extract or a compound isolated therefrom. The lycopodiella cernua extract or the compound isolated therefrom has an excellent effect of inhibiting acetylcholinesterase, butyrylcholinesterase, and β-secretase 1, and thus can be effectively used as a composition for preventing or treating Alzheimer's disease.

Description

Composition for the prophylaxis or treatment of Alzheimer's disease, comprising the extract of the Limestone River or a compound isolated therefrom

The present invention relates to a composition for the prophylaxis or treatment of Alzheimer's disease comprising an extract of Lycopodiella cernua or a compound isolated therefrom.

The rapid development of life sciences and medicine has led to an increase in the average life span of humans and the growing proportion of elderly people. In particular, senile neurological diseases such as stroke, Alzheimer's disease (AD), and Parkinson's disease (PD) appear as lethal dysfunction of the nervous system, and until now there is no effective way to prevent them. Degradation and huge medical expenses are putting a strain on the families around them.

In particular, Alzheimer's disease is the most common type of senile neurological disease, which is the most common degenerative brain disease causing dementia, a brain disease causing memory, thinking and behavioral problems. Dementia is a general term that means severe loss of memory and other intellectual skills that interfere with daily life. Alzheimer's disease is estimated to account for 60-80% of cases of dementia. In the United States, more than five million people have Alzheimer's disease, and the number of Americans with Alzheimer's disease and other dementia is expected to grow each year as the proportion of people age 65 and older in the United States continues to grow.

The pathogenesis and cause of Alzheimer's disease are not yet known. To date, the synthesis of the neurotransmitter acetylcholine, the reduction of β-amyloid, the tau protein superphosphate Nerve cell damage due to anger is known to be a major cause (Lee Eun-hee et. Al., 2008).

Acetylcholine is one of the neurotransmitters and is found mostly in the area where neurons and skeletal muscle meet. In addition, acetylcholine secreted from the nerve terminal is inactivated by decomposing into choline and acetic acid by acetylcholinesterase (AChE) after the transmission of the stimulus, the choline is an enzyme according to the action of cholineacetylase (cholinacetylase) Synthetically synthesized to form acetylcholine. Acetylcholinesterase inhibitors support signaling between neurons by preventing and maintaining high levels of acetylcholine, an important chemical messenger for memory and learning. Recent studies have also shown that butyrylcholinesterase (BChE) breaks down acetylcholine in the brain and plays a role in choline delivery. Therefore, the inhibition of the acetylcholinesterase and butyrylcholinesterase increases the concentration of acetylcholine in the synaptic gap, enhances neurotransmission of acetylcholine, and improves defects.

To date, the basic treatment for Alzheimer's disease has not been developed, but most of the therapeutic agents used in countries are acetylcholinesterase inhibitors, which cannot completely prevent the progression of the disease, and alleviate or slow down some pathological symptoms. Only effect. Drugs of this class include donepezil, rivastingmine, galantamine, and tacrine. In the development of therapeutics, according to the research trend that acetylcholinesterase is the most pharmacological action point in the world, it is possible to develop drugs and functional materials of acetylcholinesterase inhibitor with few side effects and excellent effects using various natural products. There is a need.

In addition, β-secretase 1 acts on the amyloid-β-precursor protein (APP) to generate β-amyloid fragments. In addition, since the accumulation of β-amyloid is known as a major cause of Alzheimer's disease, the development of a substance for the inhibitory activity of β-secretase may be helpful in the treatment of Alzheimer's disease by preventing the cleavage of the β-amyloid precursor protein. have. In addition, in the case of β-secretase inhibitor x-ray crystallography (x-ray crystallography) revealed a clear structure, a variety of inhibitors can be easily developed, but there is no efficacy and still available as a therapeutic agent to be. Therefore, the development of new β-secretase inhibitors could be of great help in the treatment of Alzheimer's disease.

The Lycopodiella cernua is a river of vascular plants that make spores, belonging to the family of cypresses and similar plants. It is widely spread around the world, and consists of four genera and about 1,000 species. Plants in this river are small in size and can grow upright or even grow on other plants. The stem is divided into two branches and pruned. The differentiation of roots, stems, and leaves is distinct, and photosynthesis is possible. The spore sac runs on a special leaf called the spores, which gather together to form a cone-shaped spores. It is an evergreen perennial plant, which inhabits Jeju, Jeonnam, Hambuk, Hamnam, and Gangwon.

In China, travertine has traditionally been used to treat rheumatism, whooping cough, hepatitis, kidney stones and bruises (Zhang, XC et. Al., 2004; Xiao, PG, 2002). It was also used. In addition, Korean Laid-Open Patent Publication No. 2011-0118145 suggested that lycopoduim clavatum extract or a compound isolated therefrom is effective in treating cancer, and the anti-memory loss effect of vegetable base isolated from lychee (Chuong, NN et al., 2014), anticholinesterase activity (Konrath, EL et al., 2013) of vegetable bases isolated from South American pine family has been disclosed. However, it is not yet known that the compounds isolated from the pine tree of the present invention have an inhibitory effect of acetylcholinesterase or butyrylcholinesterase or β-secretase 1.

It is an object of the present invention to provide a composition for the prevention or treatment of Alzheimer's disease, including the extract of the pine needles or a compound isolated therefrom.

The present invention relates to a composition for the prophylaxis or treatment of Alzheimer's disease comprising an extract of Lycopodiella cernua or a compound isolated therefrom. More specifically, lambnagin (Compound 1), quercetin 3,3'- dimethyl ether (Compound 2), guaiacylglycerol (Compound 3), apigenin (Compound 4), isomorphoside (Compound 1) 5), Bahia genin -4'- O - (2 '', 6 '' - di-p -O- - Kumar in one) -β-D- gluconic nose Llano side (compound 6), spring Basin -4- O -β- D - gluconic nose Llano side (compound 7), (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β- D- gluconic nose Llano side) 1'-benzofuran propionic acid (compound 8), Bahia genin -4'- O - (6 '' - O - p - in one) -β-D- gluconic nose Llano side Kumar ( Compound 9), a stone comprising at least one compound selected from the group consisting of dihydrodihydroconiferyl alcohol-4- O- β-D-glucopyranoside (compound 10) and sharposide (compound 11) Songjiang (Lycopodiella cernua) composition for preventing or treating Alzheimer's disease comprising the extract Relate to.

[Formula 1]

The travertine extract is characterized in that the extract is extracted with at least one solvent selected from the group consisting of water, C1 to C4 lower alcohol, acetone, n-hexane, dichloromethane and ethyl acetate.

The present invention is isolated from the Lycopodiella cernua ( Lycopodiella cernua ) of Rhamnajin (Compound 1), Quercetin 3,3'- dimethyl ether (Compound 2), Guaiacylglycerol (Compound 3), Apigenin (Compound 4) ), Isoshaphtoside (compound 5), apigenin-4'- O- (2 '', 6 ''-di-O- p -coumaroyl) -β-D-glucopyranoside (compound 6) ), Bombacin-4- O- β- D -glucopyranoside (Compound 7), (7 R , 8 S ) -7,8-dihydro-8-hydroxymethyl-7- (3-methoxy Phenyl-4- O- β-D-glucopyranoside) -1'-benzofuran propionic acid (Compound 8), apigenin-4'- O- (6 '' -O - p -coumaroyl) -β Selected from the group consisting of -D-glucopyranoside (Compound 9), dihydrodihydroconiferyl alcohol-4- O- β-D-glucopyranoside (Compound 10) and shaftoside (Compound 11) A composition for the prevention or treatment of Alzheimer's disease comprising at least one compound A.

The compound is characterized by inhibiting at least one enzyme activity selected from the group consisting of acetylcholinesterase, butyrylcholinesterase and β-secretase 1.

In addition, the present invention is the lambnagin (Compound 1), quercetin 3,3'- dimethyl ether (Compound 2), guaiacylglycerol (Compound 3), apigenin (Compound 4) of the formula 1 isolated from the trachea Isomorphoside (compound 5), apigenin-4'- O- (2 '', 6 ''-di-O- p -coumaroyl) -β-D-glucopyranoside (compound 6), Basin spring -4- O -β- D - gluconic nose Llano side (compound 7), (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl- 4- O- β-D-glucopyranoside) -1'-benzofuranpropionic acid (Compound 8), apigenin-4'- O- (6 '' -O - p -coumaroyl) -β-D 1 type selected from the group consisting of -glucopyranoside (Compound 9), dihydrodihydroconiferyl alcohol-4- O- β-D-glucopyranoside (Compound 10) and sharposide (Compound 11) of lycopodiopsida (Lycopodiella cernua) Alzheimer's disease comprising the extract containing the above compound It relates to a dietary supplement for a room or improvement.

In yet another aspect, the present invention comprises the steps of preparing the extract of the Songseok River with Lycopodiella cernua in water, C1 to C4 lower alcohol or a mixed solvent thereof; Fractionating the travertine extract using n-hexane and ethyl acetate in sequence and re-fractionating the water layer residue with dichloromethane; And chromatographing the water layer except for the re-distilled dichloromethane to lambnazin of the formula (compound 1), quercetin 3,3'-dimethyl ether (compound 2), guaiacylglycerol (compound 3), apigenin ( Compound 4), isomorphoside (compound 5), apigenin-4'- O- (2 '', 6 ''-di-O- p -coumaroyl) -β-D-glucopyranoside ( compound 6), spring Basin -4- O -β- D - gluconic nose Llano side (compound 7), (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3 Methoxyphenyl-4- O- β-D-glucopyranoside) -1'-benzofuranpropionic acid (compound 8), apigenin-4'- O- (6 '' -O - p -coumaroyl) In the group consisting of -β-D-glucopyranoside (Compound 9), dihydrodihydroconiferyl alcohol-4- O -β-D-glucopyranoside (Compound 10) and shaftoside (Compound 11) A method for separating one or more compounds selected.

The present invention also provides (7 R, 8 S) is a novel compound, having the following chemical structure 7,8-dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D -Glucopyranoside) -1'-benzofuran propionic acid.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for the prevention or treatment of Alzheimer's disease comprising the compound of formula (I) isolated from the Lycopodiella cernua .

The travertine extract may be obtained by extracting the travertine with water, a lower alcohol of C1 to C4 or a mixed solvent thereof, and the lower alcohol of C1 to C4 may use methanol, ethanol, propanol, isopropanol, butanol, and the like. have. In addition, the extract may be fractions re-fractionated with an organic solvent, n-hexane, ethyl acetate and dichloromethane may be used as the organic solvent.

The compound separated from the travertine can be obtained by fractionating the water layer extract of the travertine, and the chromatography is silica gel column chromatography, HP-20 column chromatography. , RP-18 column chromatography, LH-20 column chromatography, high-performance liquid chromatography, and the like.

Meanwhile, the compound of the present invention may be synthesized according to a conventional method in the art, and may be prepared as a pharmaceutically acceptable salt.

The present invention also provides a pharmaceutical composition for the prevention or treatment of Alzheimer's disease comprising at least one compound selected from the group of compounds of the formula (1) isolated from the extract of the trachea. Pharmaceutical compositions comprising the extracts or compounds are formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be used. Carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, or the like in the extract or compound of the present invention. Or it is prepared by mixing lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The dosage of the pharmaceutical composition comprising the extract of the pine needles of the present invention or a compound isolated therefrom is determined by the age, sex, weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the It will depend on your judgment. Dosage determination based on these factors is within the level of skill in the art and generally dosages range from 0.01 mg / kg / day to approximately 2000 mg / kg / day. More preferred dosage is 1 mg / kg / day to 500 mg / kg / day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

Pharmaceutical compositions comprising the extract of the pine needles of the present invention or a compound isolated therefrom can be administered to mammals such as rats, livestock, humans and the like by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections. The extract of the present invention or a compound isolated therefrom has little toxicity and side effects, and thus is a drug that can be used safely even for prolonged administration for the purpose of prevention.

In another aspect, the present invention provides a health functional food for preventing or improving Alzheimer's disease comprising at least one compound selected from the group of the compound of formula 1 isolated from the pineapple extract or the above-mentioned do. The extract or compound may be added as 0.001 to 100% by weight to the health functional food of the present invention. The health functional food of the present invention includes the form of tablets, capsules, pills, or liquids, and the food to which the extract or compound of the present invention may be added, for example, various foods, beverages, gums, teas, Vitamin complexes.

The present invention relates to a composition for the prevention or treatment of Alzheimer's disease, including the extract of the Songseok River or a compound isolated therefrom, wherein the extract of the Songseok River or the compound isolated therefrom is an acetylcholinesterase, butyrylcholinesterase and β-ce Excellent inhibitory effect of Cretease 1 can be usefully used as a composition for the prevention or treatment of Alzheimer's disease.

Figure 1 is a novel compound of (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose pyrano of the invention side) is an illustration showing a 1'-benzofuran propionic acid ^{^{1 H- 1 H COSY (bold lines}} ) and the ¹ H- ¹³ C HMBC correlation (arrow) on.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the information provided herein is to be thorough and complete, and to fully convey the spirit of the present invention to those skilled in the art.

Lycopodium cernua used in the present invention was obtained in Lumdong Province, Vietnam in April 2012, extracting and filtering the reflux of the Songsong River (3 kg) with methanol (2 L × 3 times), and filtering the filtrate. Distillation under reduced pressure gave 280 g of methanol extract. Then, the methanol extract was suspended in water (2 L) and sequentially partitioned into n-hexane (2 L × 3 times) and ethyl acetate (2 L × 3 times), and the n-hexane fraction (48 g) and ethyl acetate fraction (175.5). g) and finally a fraction (55 g) containing the water layer residue.

Thereafter, the fraction containing the water layer residue was suspended with 5% [v / v] hydrochloric acid, and then fractionated with dichloromethane to obtain a dichloromethane fraction (1.9 g). In addition, the aqueous layers of the fractions with dichloromethane to pH11-12 with 0.1N sodium hydroxide, and then fractionated with dichloromethane. The organic layer was collected, dried over sodium sulfate, and evaporated to obtain a crude alkaloid fraction. The residue containing the water layer was anionic HP- according to the concentration gradient elution condition of aqueous methanol solution (0 → 100% [v / v]). Fractionation by 20 column chromatography gave 12 fractions (W1-W12). W3 (1.24 g) in the fractions was purified by silica gel column chromatography (column size: 3.5 × 50 cm) according to isocratic elution conditions of dichloromethane: methanol: water (4: 1: 0.2 [v: v: v]). Fractionation gave 13 small fractions (W3.1-W3.13).

W3.4 (115 mg) in the small fractions was subjected to silica gel column chromatography (column size: 1.5 × 25) according to isocratic elution conditions of dichloromethane: methanol: water (5: 1: 0.1 [v: v: v]). Fractions) to give compound 3 (8.8 mg). The small fraction W3.6 (65 mg) was purified by silica gel column chromatography (column size: 1.5 × 25) according to isocratic elution conditions of dichloromethane: methanol: water (4: 1: 0.2 [v: v: v]). Fractions) to give compound 1 (28.7 mg). Small fraction W3.8 (36 mg) was prepared by RP-C18 according to the isocratic elution conditions of 20% [v / v] acetonitrile aqueous solution. Compound 2 (7.7 mg) was obtained by silica gel column chromatography.

Fraction W6 (1.05 g) was fractionated by silica gel column chromatography (column size: 3.0 × 50 cm) according to isocratic elution conditions of dichloromethane: methanol: water (3: 1: 0.2 [v: v: v]). Nine small fractions (W6.1-W6.9) were obtained. W6.2 (25 mg) in the small fractions afforded Compound 4 (6.2 mg) through filtration. The small fraction W6.9 (167 mg) was fractionated by Sephadex LH-20 column chromatography (column size: 2.0 × 120 cm) according to isocratic elution conditions of methanol: water (1: 1 [v: v]). Compound 5 (7 mg) and compound 6 (10.3 mg) were obtained.

W8 (1.52 g) in the fractions was purified by silica gel column chromatography (column size: 3.0 × 60 cm) according to isocratic elution conditions of dichloromethane: methanol: water (3: 1: 0.1 [v: v: v]). To obtain eight small fractions (W8.1-W8.8). W8.6 (125 mg) in the small fractions was separated by Sephadex LH-20 column chromatography (column size: 2.0 × 60 cm) according to isocratic elution conditions of methanol: water (1: 1 [v: v]). Fractionation gave compound 7 (3 mg). The small fraction W8.8 (150 mg) was subjected to Sephadex LH-20 column chromatography (column size: 1.5 × 120 cm) according to isocratic elution conditions of methanol: water (1: 1 [v: v]). Compound 8 (12.3 mg) was isolated.

Fraction W9 (1.36 g) was purified by silica gel column chromatography (column size: 3.0 × 60 cm) according to isocratic elution conditions of dichloromethane: methanol: water (3: 1: 0.1 [v: v: v]). Small fractions (W9.1-W9.7) were isolated. W9.4 (45 mg) in the small fractions was separated by Sephadex LH-20 column chromatography (column size: 1.5 × 120 cm) according to isocratic elution conditions of methanol: water (2: 1 [v: v]). Fractionation gave compound 9 (6.7 mg). In addition, small fraction W9.7 (65.7 mg) was purified by Sephadex LH-20 column chromatography (column size: 1.5 × 120 cm) according to isocratic elution conditions of methanol: water (2: 1 [v: v]). Fractionation gave compound 10 (14.1 mg) and compound 11 (10.5 mg).

<실시예 2. 석송강으로부터 분리된 화합물의 물리화학적 구조 확인><Example 2. Identification of Physicochemical Structure of Compound Isolated from Limestone Steel>

실시예 2-1. 람나진(화합물 1)Example 2-1. Ramnazin (Compound 1)

rhamnazin;

Yellowish powder;

IR (KBr) v _max 3545, 3400, 1658, 1130, 1117 cm ^-1 ;

¹ H NMR and ¹³ C NMR data see Table 1 and Table 2 below;

EI-MS [M] ⁺ 330.2 m / z, C ₁₇ H ₁₄ O ₇ .

실시예 2-2. 케르세틴 3,3'-디메틸 에테르(화합물 2)Example 2-2. Quercetin 3,3'-dimethyl ether (Compound 2)

quercetin 3,3'-dimethyl ether;

Yellowish powder;

¹ H NMR and ¹³ C NMR data see Table 1 and Table 2 below;

EI-MS [M] ⁺ 330.2 m / z, C ₁₇ H ₁₄ O ₇ .

실시예 2-3. 구아이아실글리세롤(화합물 3)Example 2-3. Guaiacylglycerol (Compound 3)

guaiacylglycerol;

White powder;

¹ H NMR and ¹³ C NMR data see Table 1 and Table 2 below;

EI-MS [M] ⁺ 214.2 m / z, C ₁₀ H ₁₄ O ₅ .

실시예 2-4. 아피게닌(화합물 4)Example 2-4. Apigenin (Compound 4)

apigenin;

Yellowish powder;

¹ H NMR and ¹³ C NMR data see Table 1 and Table 2 below;

EI-MS [M] ⁺ 270.0 m / z , C ₁₅ H ₁₀ O ₅ .

실시예 2-5. 이소샤프토사이드(화합물 5)Example 2-5. Isomorphoside (Compound 5)

isoschaftoside;

Yellowish powder;

¹ H NMR and ¹³ C NMR data see Table 3 below;

EI-MS [M] ⁺ 564.4 m / z, C ₂₆ H ₂₈ O ₁₄ .

실시예 2-6. 아피게닌-4'-Example 2-6. Apigenin-4'- OO -(2'',6''-디-O--(2 '', 6 ''-D-O- pp -쿠마로일)-β-D-글루코피라노사이드(화합물 6)-Coumaryl) -β-D-glucopyranoside (Compound 6)

apigenin-4'- O- (2 '', 6 ''-di- O - p- coumaroyl) -β-D-glucopyranoside;

White amorphous powder;

¹ H NMR and ¹³ C NMR data see Table 4 below;

ESI-MS [M + H] 579.1 m / z, C ₃₀ H ₂₇ O ₁₂ .

실시예 2-7. 봄바신-4-Example 2-7. Bombasin-4- OO -β--β- DD -글루코피라노사이드(화합물 7)Glucopyranoside (Compound 7)

bombasin-4- O- β- D- glucopyranoside;

White amorphous powder;

UV (MeOH): 231 (4.23), 283 (4.17) nm;

IR (KBr): 3367, 2920, 1651, 1601, 1514, 1464, 1427, 1265, 1223, 1159, 1126 cm ^-1 ;

¹ H NMR and ¹³ C NMR data see Table 4 below;

EI-MS [M] ⁺ 506.18 m / z, C ₂₅ H ₃₀ O ₁₁ .

실시예 2-8. (7Example 2-8. (7 RR ,8,8 SS )-7,8-디히드로-8-히드록시메틸-7-(3-메톡시페닐-4-) -7,8-dihydro-8-hydroxymethyl-7- (3-methoxyphenyl-4- OO -β-D-글루코피라노사이드)-1'-벤조푸란프로피온산(화합물 8)-β-D-glucopyranoside) -1'-benzofuran propionic acid (compound 8)

(7R, 8S) -7,8-dihydro-8-hydroxymethyl-7- (3-methoxyphenyl-4-O-β-D-glucopyranoside) -1'-benzofuranpropanoic acid (also called "lycocernuaside A")

White amorphous powder;

-36.2 ° ( c 0.53, MeOH);

IR (KBr) ν _max 3379, 2941, 2834, 1638, 1600, 1515, 1453, 1274, 1032 cm ^-1 ;

UV (MeOH) λ _max (log ε ) 281 (2.50), 230 (3.12) nm;

ECD ( c 0.030 mM, MeOH): Δ ε ₂₇₈ -2.38, Δ ε ₂₃₂ -1.42, Δ ε ₂₁₆ +1.30;

¹ H NMR and ¹³ C NMR data see Table 5 below;

HR-FAB-MS m / z 559.1794 [M + Na] ⁺ (calcd for C ₂₆ H ₃₂ O ₁₂ Na, 559.1791).

실시예 2-9. 아피게닌-4'-Example 2-9. Apigenin-4'- OO -(6''--(6 ''- OO -- pp -쿠마로일)-β-D-글루코피라노사이드 (화합물 9)-Coumaryl) -β-D-glucopyranoside (Compound 9)

apigenin-4'- O- (6 '' -O - p- coumaroyl) -β-D-glucopyranoside;

White amorphous powder;

¹ H NMR and ¹³ C NMR data see Table 6 below;

ESI-MS [M + H] 579.1 m / z, C ₃₀ H ₂₇ O ₁₂ .

실시예 2-10. 디히드로디히드로코니페릴알코올-4-Example 2-10. Dihydrodihydroconiferyl Alcohol-4- OO -β-D-글루코피라노사이드(화합물 10)-β-D-glucopyranoside (Compound 10)

dihydrodehydroconiferyl alcohol-4- O- β-D-glucopyranoside;

Colorless rubber;

IR (KBr) v _max 3380, 2945, 2834, 1600, 1517, 1462, 1270, 1035 cm ^-1 ;

UV (MeOH) λ _max (logε) 280 (2.35), 232 (3.0) nm;

¹ H NMR and ¹³ C NMR data see Table 6 below;

EI-MS m / z 552 [M] ⁺ (calcd for C ₂₆ H ₃₄ O _11, 552).

실시예 2-11. 샤프토사이드(화합물 11)Example 2-11. Sharptoside (Compound 11)

schaftoside;

Yellowish powder;

¹ H NMR and ¹³ C NMR data see Table 3 below;

EI-MS [M] ⁺ 564.4 m / z, C ₂₆ H ₂₈ O ₁₄ .

Table 1

	Compound 1	Compound 2	Compound 3	Compound 4
Position	δ _H (ppm) ^a	δ _H (ppm) ^a	δ _H (ppm) ^b	δ _H (ppm) ^a
One			4.43 (1H, doublet, J = 6.4 μs)
2			3.59 (1H, m, H-2)
3			3.41 (1H, doublet of doublets, J = 11.2, 4.0 Hz, H-3a); 3.27 (1H, m, H-3b)	6.68 (1 H, s)
4
5
6	6.77 (1H, s, J = 2.0 μs)	6.75 (1H, s, J = 2.0 μs)		6.54 (1H, s, J = 2.0 μs)
7
8	6.38 (1H, s, J = 2.0 μs)	6.40 (1H, s, J = 2.0 μs)		6.25 (1H, s, J = 2.0 μs)
9
10
One'
2'	7.68 (1H, doublet, J = 2.0 μs)	7.70 (1H, doublet, J = 2.0 μs)	6.91 (1H, doublet, J = 1.2 H)	7.98 (1H, doublet, J = 8.7 μs)
3 '				7.20 (1H, doublet, J = 8.7 μs)
4'
5 '	6.92 (1H, s, J = 8.0 μs)	6.92 (1H, s, J = 8.0 μs)	6.69 (1H, doublet, J = 8.0 μs)	7.20 (1H, doublet, J = 8.7 μs)
6 '	7.62 (1H, doublet of doublets, J = 8.0, 2.0 μs)	7.62 (1H, doublet of doublets, J = 8.0, 2.0 μs)	6.73 (1H, doublet of doublets, J = 8.0, 1.2 μs)	7.98 (1H, doublet, J = 8.7 μs)
7-MeO	3.86 (3H, s)
3-MeO		3.80 (3H, s)
3'-MeO	3.87 (3H, s)	3.85 (3H, s)	3.78 (3H, s)
5-OH	12.05 (1H, s)	12.14 (1 H, s)
a: ¹ H NMR (400 MHz in DMSO-d ₆ ) spectroscopy data; b: ¹ H NMR (400 MHz in MeOD-d ₄ ) spectroscopy data.

TABLE 2

	Compound 1	Compound 2	Compound 3	Compound 4
Position	δ _C (ppm) ^a	δ _C (ppm) ^a	δ _C (ppm) ^b	δ _C (ppm) ^a
One			75.6
2	156.3	156.5	77.7	164.2
3	133.6	136.5	64.3	105
4	177.7	178.2		182.5
5	160.8	161.2		158.7
6	97.9	97.3		94.7
7	165.2	164.4		164.8
8	92.4	92.6		99.6
9	157	157.7		163.2
10	105	106.2		105.2
One'	120.7	122.5	134.9	126
2'	112.6	112.4	111.6	128.9
3 '	149.6	150.5	147.2	117.5
4'	147.2	147.7	149	161
5 '	115.4	115.6	116	117.5
6 '	122.9	121.4	120.8	128.9
7-MeO	55.4
3-MeO		55.2
3'-MeO	55.7	56.1	56.4
5-OH
a: ¹³ C-NMR (100 MHz in DMSO-d ₆ ) spectroscopy data; b: ¹³ C-NMR (100 MHz in MeOD-d ₄ ) spectroscopy data.

TABLE 3

	Compound 5		Compound 11
Position	δ _H (ppm) ^a	δ _C (ppm) ^b	Position	δ _H (ppm) ^c	δ _C (ppm) ^d
2		165.2	2		166.6
3	6.63 (1 H, s)	105.5	3	6.41 (1 H, s)	105.5
4		183.5	4		184.1
5		156.8	5		157.4
6		106.8	6		108.3
7		163.2	7		163
8		103.8	8		103.7
9		163.2	9		162.7
10		109.6	10		105.7
One'		123.5	One'		123.3
2'	8.32 (1H, doublet, J = 8.8 μs)	129.7	2'	7.80 (1H, doublet, J = 8.0 μs)	129.6
3 '	7.27 (1H, doublet, J = 8.8 μs)	117.5	3 '	7.74 (1H, doublet, J = 8.0 μs)	117.1
4'		160.5	4'		160.1
5 '	7.27 (1H, doublet, J = 8.8 μs)	117.5	5 '	7.74 (1H, doublet, J = 8.0 μs)	117.1
6 '	8.32 (1H, doublet, J = 8.8 μs)	129.7	6 '	7.80 (1H, doublet, J = 8.0 μs)	129.6
8-Glc			6-Glc
One''	5.86 (1H, doublet, J = 9.6 kPa)	75.7	One''	4.93 (1H, doublet, J = 9.6 kPa)	75.5
2''	4.68 (1H, m)	81.4	2''	3.75 (1 H, m)	82.6
3 ''	4.39 (1 H, m)	76.7	3 ''	3.72 (1H, m)	80.3
4''	4.28 (1 H, m)	71.9	4''	3.49 (1H, m)	71.5
5 ''	4.50 (1 H, m)	72.8	5 ''	3.60 (1H, m)	72.4
6 ''	4.57-4.28 (1H, m)	63.4	6 ''	3.32 (1H, m), 3.84 (1H, m)	63.2
6-Rha			8-Rha
One'''	5.66 (1H, doublet, J = 9.6 kPa)	84	One'''	4.63 (1H, doublet, J = 9.6 kPa)	82.9
2'''	4.67 (1 H, m)	76.2	2'''	3.90 (1H, m)	76.6
3 '''	4.48 (1 H, m)	73.4	3 '''	3.75 (1 H, m)	73.2
4'''	4.37 (1 H, m)	71.4	4'''	3.42 (1 H, m)	71.2
5 '''	4.33 (1 H, m)	72.2	5 '''	3.80 (1 H, m)	71.7
a: ¹ H NMR (400 MHz in Pyridine- d ₅ ) spectroscopy data; b: ¹³ C-NMR (100 MHz in Pyridine- d ₅ ) spectroscopy data; c: ¹ H NMR (400 MHz in DMSO- d ₆ ) spectroscopy data; d: ¹³ C-NMR (100 MHz in DMSO- d ₆ ) spectroscopy data.

Table 4

	Compound 6		Compound 7
Position	δ _H (ppm) ^a	δ _C (ppm) ^b	δ _H (ppm) ^c	δ _C (ppm) ^d
One				130.8
2		163.8	7.33 (1H, doublet, J = 2.0 μs)	111.9
3	6.71 (1 H, s)	105.7		150.6
4		182.6		148.4
5		158.3	7.19 (1H, doublet of doublets, J = 2.0, 8.5 Hz)	116.6
6	6.53 (1H, s, J = 2.0 μs)	94.5	7.59 (1H, doublet, J = 8.5 μs)	119.5
7		165.1	6.16 (1H, doublet, J = 6.5 μs)	89.3
8	6.27 (1H, s, J = 2.0 μs)	100.2	3.96 (1H, m)	54.2
9		163.7	4.27 (2H, m)	64
10		105.1
One'		124.6	7.78 (1 H, brs)	120.1
2'	7.90 (1H, doublet, J = 8.7 μs)	129.1		132.6
3 '	7.18 (1H, doublet, J = 8.7 μs)	117.5	7.92 (1 H, brs)	113.6
4'		161.3		137.6
5 '	7.18 (1H, doublet, J = 8.7 μs)	117.4		147
6 '	7.90 (1H, doublet, J = 8.7 μs)	129.1		145.2
7 '				196.7
8'			2.58 (3H, s)	26.5
9 '
One''	5.42 (1H, doublet, J = 8.1 μs)	99.8	5.69 (1H, doublet, J = 7.0 μs)	102.7
2''	5.15 (1H, doublet of doublets, J = 9.4, 8.1 Hz)	74.2	4.36 (1H, m)	75
3 ''	3.80 (1H, t, J = 9.0 μs)	73.9	4.37 (1 H, m)	78.6
4''	3.70 (1H, t, J = 9.0 μs)	77.3	4.32 (1H, m)	71.4
5 ''	3.78 (1 H, m)	74.6	4.13 (1 H, m)	79
6 ''	3.83 (1H, doublet of doublets, J = 12.1, 2.0 Hz); 3.74 (1H, doublet of doublets, J = 12.1, 5.5 Hz)	66.4	4.55 (1H, doublet of doublets, J = 2.0, 12.0 μs); 4.37 (1 H, m)	62.5
One'''		126.7
2'''	7.54 (1H, d, J = 8.5 μs)	131.2
3 '''	6.68 (1H, doublet, J = 8.5 μs)	116.5
4'''		160.8
5 '''	6.68 (1H, doublet, J = 8.5 μs)	115.4
6 '''	7.54 (1H, d, J = 8.5 μs)	130.5
7 '''	7.66 (1H, doublet, J = 15.0 μs)	146
8'''	6.37 (1H, doublet, J = 15.0 μs)	114.8
9 '''		166.7
One''''		125.8
2''''	7.53 (1H, doublet, J = 8.5 μs)	131.1
3 ''	6.86 (1H, doublet, J = 8.5 μs)	116.6
4''''		160.5
5 ''	6.86 (1H, doublet, J = 8.5 μs)	116.6
6 ''''	7.53 (1H, doublet, J = 8.5 μs)	131.1
7 ''''	7.60 (1H, doublet, J = 15.0 μs)	145.6
8''''	6.39 (1H, doublet, J = 15.0 μs)	114.7
9 ''''		167.8
3-OMe			3.86 (3H, s)	56.2
6'-OMe			3.69 (3H, s)	56.2
a: ¹ H NMR (400 MHz in DMSO- d ₆ ) spectroscopy data; b: ¹³ C-NMR (100 MHz in DMSO- d ₆ ) spectroscopy data; c: ¹ H NMR (400 MHz in MeOD- d ₄ ) spectroscopy data; d: ¹³ C-NMR (100 MHz in MeOD- d ₄ ) spectroscopy data.

Table 5

	Compound 8
Position	δ _H (ppm, J in ㎐) ^a	δ _C (ppm) ^b
One		129.6
2	7.06, brs	111.2
3		150.9
4		147.5
5	7.17, d (8.0)	118.1
6	6.96, brd (8.0)	119.4
7	5.55, d (6.0)	88.4
8	3.51, m	55.5
9	3.88, m; 3.78, m	65.2
One'		136.1
2'	6.80, brs	114.1
3 '		138.5
4'		147.7
5 '		145.2
6 '	6.76, brs	117.8
7 '	2.84, t (7.6)	32.3
8'	2.53, t (7.6)	38.1
9 '		178.1
One''	4.89, d (7.6)	102.8
2''	3.53, m	74.8
3 ''	3.51, m	77.8
4''	3.43, m	71.3
5 ''	3.45, m	79.4
6 ''	3.72, dd (3.5, 12.0); 3.89, m	62.4
3-OMe	3.88, s	56.7
5'-OMe	3.85, s	56.8
a: ¹ H NMR (500 MHz in Methanol- d ₄ , d values) spectroscopic data; b: ¹³ C NMR (125 MHz in Methanol- d ₄ , d values) spectroscopic data.

Table 6

	Compound 9		Compound 10
Position	δ _H (ppm) ^a	δ _C (ppm) ^b	δ _H (ppm) ^c	δ _C (ppm) ^d
One				129.4
2		164.2	7.01 (1H, broad singlet)	111.3
3	6.68 (1 H, s)	105		150.7
4		182.5		147.4
5		158.7	6.87 (1H, doublet, J = 8.5 μs)	117.9
6	6.54 (1H, s, J = 2.0 μs)	94.7	6.99 (1H, doublet of doublets, J = 1.5, 8.5 Hz)	119.2
7		164.8	5.61 (1H, d, 6.0 Hz)	88.3
8	6.25 (1H, s, J = 2.0 μs)	99.6	3.55 (1H, m)	55.4
9		163.2	3.91 (1 H, m); 3.82 (1 H, m)	64.9
10		105.2
One'		126	6.79 (1H, brs)	120.3
2'	7.98 (1H, doublet, J = 8.7 μs)	128.9		132.7
3 '	7.20 (1H, doublet, J = 8.7 μs)	117.5	7.20 (1H, brs)	114.9
4'		161		138.2
5 '	7.20 (1H, doublet, J = 8.7 μs)	117.5		147.3
6 '	7.98 (1H, doublet, J = 8.7 μs)	128.9		145
7 '			2.69 (2H, t, J = 7.5 μs)	32.6
8'			1.88 (2H, m)	36
9 '			3.63 (2H, t, J = 6.5 μs)	62.3
One''	5.39 (1H, doublet, J = 8.1 μs)	99.4	4.95 (1H, doublet, J = 7.0 μs)	102.6
2''	5.17 (1H, doublet of doublets, J = 9.4, 8.1 Hz)	74.1	3.53 (1 H, m)	74.7
3 ''	3.82 (1H, t, J = 9.0 μs)	75.6	3.45 (1H, m)	77.6
4''	3.61 (1H, t, J = 9.0 μs)	71.2	3.46 (1H, m)	71.1
5 ''	3.71 (1 H, m)	78.1	3.52 (1 H, m)	77.9
6 ''	3.96 (1H, doublet of doublets, J = 12.1, 2.0 μs); 3.76 (1H, doublet of doublets, J = 12.1, 5.8 Hz)	62.2	3.74 (1H, doublet of doublets, J = 3.5, 12.0 μs); 3.94 (1 H, m)	62
One'''		126.8
2'''	7.54 (1H, d, J = 8.5 μs)	130.9
3 '''	6.68 (1H, doublet, J = 8.5 μs)	116.5
4'''		160.5
5 '''	6.68 (1H, doublet, J = 8.5 μs)	116.5
6 '''	7.54 (1H, d, J = 8.5 μs)	130.9
7 '''	7.66 (1H, doublet, J = 15.0 μs)	145.5
8'''	6.37 (1H, doublet, J = 15.0 μs)	115.2
9 '''		166.5
3-OMe			3.90 (3H, s)	56.5
6'-OMe			3.94 (3H, s)	56.6
a: ¹ H NMR (400 MHz in DMSO- d ₆ ) spectroscopy data; b: ¹³ C NMR (100 MHz in DMSO- d ₆ ) spectroscopy data; c: ¹ H NMR (400 MHz in MeOD- d ₄ ) spectroscopy data; d: ¹³ C NMR (100 MHz in MeOD- d ₄ ) spectroscopy data.

<실시예 3. 아세틸콜린에스테라아제 및 부티릴콜린에스테라아제의 억제활성 측정>Example 3 Determination of Inhibitory Activity of Acetylcholinesterase and Butyrylcholinesterase

Hydrolysis of acetylcholine or butyrylacetylcholine by cholinesterase results in the formation of a yellow 5'-thio-2-nitrobenzoate anion (5'-thio-2-nitrobenzoate anion). The enzyme activity was measured through.

140 µl of sodium phosphate buffer (pH 8.0) and 20 µl of each compound of the present invention with a final concentration of 100 µM, acetylcholinesterase or butyrylcholinesterase (5 units / ml) in 96 wells microplates 20 µl, 10 µl of dithiobisnitrobenzoate (DTNB), 10 µl of acetylcholine or butyrylcholine were added and reacted at room temperature for 15 minutes. As a positive control group, berberine was used, and the 5'-thio-2-nitrobenzoate anion formed after the reaction was confirmed at 412 nm with an absorbance meter (VERSA max, Molecular Devices, USA). ₅₀ (the half maximal inhibitory concentration).

TABLE 7

Condition	Acetylcholinesterase inhibitory activity (IC ₅₀ )	Butyrylcholinesterase Inhibitory Activity (IC ₅₀ )
Compound 1	8.69 ± 0.49μM	> 30 μM
Compound 2	8.78 ± 0.97μM	13.04 ± 0.99 μM
Compound 3	1.56 ± 0.22μM	> 30 μM
Compound 4	7.29 ± 0.23μM	15.78 ± 0.95 μM
Compound 5	0.23 ± 0.17μM	3.08 ± 0.26μM
Compound 6	1.24 ± 0.31μM	3.11 ± 0.77μM
Compound 7	11.25 ± 0.99μM	12.23 ± 0.89μM
Compound 8	15.22 ± 0.89μM	24.09 ± 0.33μM
Compound 9	2.78 ± 0.42μM	7.15 ± 0.19μM
Compound 10	11.93 ± 0.96μM	7.02 ± 0.29μM
Compound 11	1.70 ± 0.22μM	0.62 ± 0.47μM
Berberine	0.71 ± 0.11μM	7.08 ± 0.01μM

Referring to the results of Table 7, it was confirmed that the compound isolated from the extract of the pine tree of the present invention has an inhibitory activity of acetylcholinesterase or butyrylcholinesterase, which can be usefully used as a prophylactic and therapeutic agent for Alzheimer's disease Can be.

<실시예 4. β-세크레타제 1의 억제활성 측정>Example 4 Determination of Inhibitory Activity of β-Secretase 1

Fluorescence was generated when the substrate was cut by β-secretase 1, and the enzyme activity was confirmed by measuring it with a BACE1 FRET assay kit (β-Secretase, human recombinant, PanVera Co. USA).

To each 10 μl of 50 mM sodium acetate (pH4.5), β-secretase 1 and the substrate (750 nM Rh-EVNLDAEFK-Quencher), 10 μl of each compound was added and reacted in the dark at 25 ° C. for 60 minutes. . Quercetin was used as a positive control, here measuring fluorescence at the wavelength and emission wavelength 545㎚ 585㎚ to determine the reaction results, it is shown to the _{IC 50 (the half maximal inhibitory concentration} ) in Table VIII.

Table 8

Condition	β-secretase 1 inhibitory activity (IC ₅₀ )
Compound 1	5.59 ± 0.33μM
Compound 3	7.33 ± 1.77μM
Compound 10	26.86 ± 1.94μM
Compound 11	2.16 ± 0.37μM
Quercetin	6.76 ± 0.63μM

Looking at the results of Table 8, it was confirmed that the compound isolated from the extract of the pine tree of the present invention inhibits the activity of β-secretase 1, it can be confirmed that the therapeutic effect for Alzheimer's disease is excellent.

<실시예 5. 독성실험> Example 5. Toxicity Test

실시예 5-1. 급성독성 Example 5-1. Acute Toxicity

Of the present invention lycopodiopsida extract or (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) This experiment was carried out to investigate the toxicity of acutely (within 24 hours) to the body of an animal when a large amount of -1'-benzofuran propionic acid (Compound 1) was ingested and to determine the lethality. Twenty ICR mice, which are common mice, were prepared, and 10 were assigned to each group. Only 30% PEG-400 was administered to the control group, and the experimental group orally administered the extract or compound 1 of the present invention at a concentration of 1.0 g / kg, respectively. Each mortality rate was examined 24 hours after administration, and both the control group and the experimental group administered with the extract or Compound 1 survived.

실시예 5-2. 실험군 및 대조군의 장기 및 조직 독성 실험Example 5-2. Long-term and tissue toxicity experiments in experimental and control groups

Long-term toxicity experiments lycopodiopsida extract or (7 R, 8 S) of the present invention to investigate the effect of each organ (tissue) of the animals intended for C57BL / 6J mice, 7,8-dihydro-8-hydroxy Of the experimental group in which oxymethyl-7- (3-methoxyphenyl-4- O- β-D-glucopyranoside) -1'-benzofuran propionic acid was administered at a concentration of 1.0 g / kg and the control group in which only the solvent was administered Blood was collected after 8 weeks from the animals, and blood concentrations of glutamate-pyruvate transferase (GPT) and blood urea nitrogen (BUN) were measured using a Select E (vital scientific NV, Netherland) instrument. As a result, in the case of GPT, which is known to be related to hepatotoxicity, and BUN, which is known to be related to nephrotoxicity, there was no difference in the experimental group compared to the control group. In addition, liver and kidneys were excised from each animal, and histological observations were performed by optical microscopy through a conventional tissue fabrication process. No abnormalities were observed.

<제제예 1. 약학적 제제>Preparation Example 1. Pharmaceutical Formulations

제제예 1-1. 정제의 제조Formulation Example 1-1. Manufacture of tablets

Of the present invention (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) 1' 200 g of benzofuran propionic acid was mixed with 175.9 g lactose, 180 g potato starch and 32 g colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into a tablet.

제제예 1-2. 주사제의 제조Formulation Example 1-2. Preparation of Injectables

Of the present invention (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) 1' 1 g of benzofuran propionic acid, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.

<제제예 2. 식품 제조>Preparation Example 2 Food Preparation

제제예 2-1. 조리용 양념의 제조Formulation Example 2-1. Preparation of Cooking Seasonings

Of the present invention (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) 1' Benzofuran propionic acid was added to the cooking seasoning at 1% by weight to prepare a cooking seasoning for health promotion.

제제예 2-2. 밀가루 식품의 제조Formulation Example 2-2. Manufacture of Flour Food

Of the present invention (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) 1' Benzofuran propionic acid was added to the flour at 0.1% by weight, and the mixture was used to prepare bread, cake, cookies, crackers and noodles to prepare foods for health promotion.

제제예 2-3. 스프 및 육즙(gravies)의 제조Formulation Example 2-3. Preparation of soups and gravy

Of the present invention (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) 1' Benzofuran propionic acid was added to the soup and broth in 0.1% by weight to prepare soup and broth for health promotion.

제제예 2-4. 유제품(dairy products)의 제조Formulation Example 2-4. Manufacture of dairy products

Of the present invention (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) 1' Benzofuran propionic acid was added to the milk at 0.1% by weight, and the milk was used to prepare various dairy products such as butter and ice cream.

제제예 2-5. 야채주스 제조Formulation Example 2-5. Vegetable Juice Manufacturing

Of the present invention (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) 1' 0.5 g of benzofuran propionic acid was added to 1,000 ml of tomato juice or carrot juice to prepare vegetable juice for health promotion.

제제예 2-6. 과일주스 제조Formulation Example 2-6. Fruit juice manufacturing

Of the present invention (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) 1' 0.1 g of benzofuran propionic acid was added to 1,000 ml of apple juice or wine to prepare fruit juice for health promotion.

Claims

Ramnazin (Compound 1), Quercetin 3,3'-dimethyl ether (Compound 2), Guaiacylglycerol (Compound 3), Apigenin (Compound 4), Isomorphoside (Compound 5), Api Genenin-4'- O- (2 '', 6 ''-di-O- p -coumaroyl) -β-D-glucopyranoside (Compound 6), bombacin-4- O- β- D - glue nose Llano side (compound 7), (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Ranoside) -1′-benzofuranpropionic acid (compound 8), apigenin-4′- O- (6 '' -O - p -coumaroyl) -β-D-glucopyranoside (compound 9), Lycopodiella cernua comprising at least one compound selected from the group consisting of dihydrodihydroconiferyl alcohol-4- O- β-D-glucopyranoside (compound 10) and sharposide (compound 11) A composition for the prevention or treatment of Alzheimer's disease, characterized in that it contains an extract.

[Formula 1]
The method of claim 1,

Said Songseok River extract is an extract of Alkheimer's disease, characterized in that the extract of the Songseok River with water, C1 to C4 lower alcohol, acetone, n-hexane, dichloromethane and ethylacetate selected from the group consisting of one or more solvents Or a therapeutic composition.
Rhamnazin (Compound 1), Quercetin 3,3'-dimethyl ether (Compound 2), Guaiacylglycerol (Compound 3), Apigenin (Compound 4), Iso isolated from Lycopodiella cernua Shaptoside (Compound 5), Apigenin-4'- O- (2 '', 6 ''-di-O- p -coumaroyl) -β-D-glucopyranoside (Compound 6), spring Basin -4- O -β- D - gluconic nose Llano side (compound 7), (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4 O- β-D-glucopyranoside) -1'-benzofuranpropionic acid (compound 8), apigenin-4'- O- (6 '' -O - p -coumaroyl) -β-D- At least one member selected from the group consisting of glucopyranoside (compound 9), dihydrodihydroconiferyl alcohol-4- O- β-D-glucopyranoside (compound 10) and sharposide (compound 11) A composition for preventing or treating Alzheimer's disease, comprising a compound.

[Formula 1]
The method according to claim 1 or 3,

Alzheimer's disease, characterized in that the compound inhibits at least one enzyme activity selected from the group consisting of acetylcholinesterase, butyrylcholinesterase and β-secretase 1 (β-secretase 1) Composition for the prevention or treatment of
Ramnazin (Compound 1), Quercetin 3,3'-dimethyl ether (Compound 2), Guaiacylglycerol (Compound 3), Apigenin (Compound 4), Isomorphoside (Compound 5), Api Genenin-4'- O- (2 '', 6 ''-di-O- p -coumaroyl) -β-D-glucopyranoside (Compound 6), bombacin-4- O- β- D - glue nose Llano side (compound 7), (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Ranoside) -1′-benzofuranpropionic acid (compound 8), apigenin-4′- O- (6 '' -O - p -coumaroyl) -β-D-glucopyranoside (compound 9), Lycopodiella cernua comprising at least one compound selected from the group consisting of dihydrodihydroconiferyl alcohol-4- O- β-D-glucopyranoside (compound 10) and sharposide (compound 11) Health functional food for the prevention or improvement of Alzheimer's disease, characterized in that it contains an extract.

[Formula 1]
The method of claim 5,

Said Songseok River extract is an extract of Alkheimer's disease, characterized in that the extract of the Songseok River with water, C1 to C4 lower alcohol, acetone, n-hexane, dichloromethane and ethylacetate selected from the group consisting of one or more solvents Or dietary supplements for improvement.
Rhamnazin (Compound 1), Quercetin 3,3'-dimethyl ether (Compound 2), Guaiacylglycerol (Compound 3), Apigenin (Compound 4), Iso isolated from Lycopodiella cernua Shaptoside (Compound 5), Apigenin-4'- O- (2 '', 6 ''-di-O- p -coumaroyl) -β-D-glucopyranoside (Compound 6), spring Basin -4- O -β- D - gluconic nose Llano side (compound 7), (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4 O- β-D-glucopyranoside) -1'-benzofuranpropionic acid (compound 8), apigenin-4'- O- (6 '' -O - p -coumaroyl) -β-D- At least one member selected from the group consisting of glucopyranoside (compound 9), dihydrodihydroconiferyl alcohol-4- O- β-D-glucopyranoside (compound 10) and sharposide (compound 11) Health functional food for the prevention or improvement of Alzheimer's disease comprising a compound.

[Formula 1]
The method according to claim 5 or 7,

Alzheimer's disease, characterized in that the compound inhibits at least one enzyme activity selected from the group consisting of acetylcholinesterase, butyrylcholinesterase and β-secretase 1 (β-secretase 1) Functional foods for the prevention or improvement of
Preparing an extract of S. limpidus ( Lycopodiella cernua ) with water, C1 to C4 lower alcohol or a mixed solvent thereof; Fractionating the travertine extract using n-hexane and ethyl acetate in sequence and re-fractionating the water layer residue with dichloromethane; And chromatographing the water layer except for the re-distilled dichloromethane with lambnazin (Compound 1), quercetin 3,3'-dimethyl ether (Compound 2), guaiacylglycerol (Compound 3), and apigenin (Formula 1). Compound 4), isomorphoside (compound 5), apigenin-4'- O- (2 '', 6 ''-di-O- p -coumaroyl) -β-D-glucopyranoside ( compound 6), spring Basin -4- O -β- D - gluconic nose Llano side (compound 7), (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3 Methoxyphenyl-4- O- β-D-glucopyranoside) -1'-benzofuranpropionic acid (compound 8), apigenin-4'- O- (6 '' -O - p -coumaroyl) In the group consisting of -β-D-glucopyranoside (Compound 9), dihydrodihydroconiferyl alcohol-4- O -β-D-glucopyranoside (Compound 10) and shaftoside (Compound 11) A method of separating one or more compounds selected.

[Formula 1]
To a novel compound having a chemical structure (7 R, 8 S) -7,8- dihydro-8-hydroxy-7- (3-methoxyphenyl -4- O -β-D- gluconic nose Llano side) -1'-benzofuran propionic acid.