KR20090048710A - Composition for improving impairment of cholinergic nervous system containing extract of anemarrhena asphodeloides or compound isolated from the same - Google Patents

Abstract

The present invention is a hair extract; One or more compounds selected from the group consisting of mangiferin, neomangiferin and timosaponin AIII; Or it relates to a choline nervous system damage improving agent comprising norathyriol (norathyriol) as an active ingredient. Hair extracts and compounds of the present invention can be usefully used for the treatment or prevention of memory loss and brain damage due to choline nervous system damage.

Hair loss, extract, choline nervous system damage improvement, memory loss treatment

Description

CHOLINERGIC NERVOUS SYSTEM CONTAINING EXTRACT OF ANEMARRHENA ASPHODELOIDES OR COMPOUND ISOLATED FROM THE SAME}

The present invention relates to choline nervous system damage improving agent comprising an extract or a compound isolated therefrom as an active ingredient. More specifically, the present invention is a hair extract; One or more compounds selected from the group consisting of mangiferin, neomangiferin and timosaponin AIII; Or it relates to a choline nervous system damage improving agent comprising norathyriol (norathyriol) as an active ingredient.

Jimo ( Anemarrhena Asphodeloides Bunge is used as a medicinal herb as dried herbs and peeled dried rhizome (Gwangjimo), and its main effects are hypoglycemic action, antipyretic action and platelet aggregation inhibition. , Diuretic, anticancer and the like are known (see references [2], [6] and [13]). About 6% of total saponin is contained in the rhizome. Among them, thymosaponin AI, thymosaponin AII, thymosaponin AIII, thymosaponin AIV, thymosaponin BI, and thymosaponin BII are known. Newly reported marsaponin F, G, and the like (see Ref. [8]). These sapogenins are mainly sarsarsapogenin, markogenin, neogitogenin, lilagenin and the like (see Ref. [8]). Xanthone-based compounds contain manziperin and neomanziferin and are known for their antidiabetic effects (see references [5], [9] and [10]). Steroid-based saponins are the main component of the hair, and sarsasapogenin, the parent of these saponins, has been demonstrated to have high antidepressant effects by Ren et al. (Ref. [12]).

To date, all of the functions related to the memory of the hair are related to the improvement effect of the abnormal symptoms caused by ischemia. Oh et al (Ref. [11]) indicate that the water extract of the hair is used to detect infarcts and edema caused by the ischemia. Li et al. (7) reported that inflammation induced by ischemia was alleviated by thymosaponin BII, improving memory and learning abnormalities. In addition, Gottlieb et al. (Ref. [4]) also demonstrated the effect of Manziferin to mitigate the effects of excitatory toxicity and free radical formation on the nervous system due to cerebral ischemia.

However, there has been no report on the improvement effect of Jimi on memory impairment due to choline nervous system abnormalities. The choline nervous system in the basal region of the forebrain is responsible for memory and cognitive function, and therefore, the decline in memory and cognitive ability is known to be due to damage of the choline nervous system in the brain due to aging (see Ref. [1]). In addition, scopolamine is an anticholinergic substance and is known to cause memory dysfunction by reducing the function of acetylcholine at synapses without affecting the concentration of acetylcholine (see Ref. [3]).

references:

[1] Beatty, W. W., Butters, N., Janowsky, D. S., 1986. Patterns of memory failure after scopolamine treatment: implications for cholinergic hypotheses of dementia. Behav Neural Biol 45, 196-211.

[2] Dong, J. X., Han, G. Y., 1991. A new active steroidal saponin from Anemarrhena asphodeloides. Planta Med 57, 460-462.

[3] Ebert, U., Kirch, W., 1998. Scopolamine model of dementia: electroencephalogram findings and cognitive performance. Eur J Clin Invest 28, 944-949.

[4] Gottlieb, M., Leal-Campanario, R., Campos-Esparza, MR, Sanchez-Gomez, MV, Alberdi, E., Arranz, A., Delgado-Garcia, JM, Gruart, A., Matute, C., 2006. Neuroprotection by two polyphenols following excitotoxicity and experimental ischemia. Neurobiology of disease 23, 374-386.

[5] Ichiki, H., Miura, T., Kubo, M., Ishihara, E., Komatsu, Y., Tanigawa, K., Okada, M., 1998. New antidiabetic compounds, mangiferin and its glucoside. Biol Pharm Bull FIELD Full Journal Title: Biological & pharmaceutical bulletin 21, 1389-1390.

[6] Iida, Y., Oh, KB, Saito, M., Matsuoka, H., Kurata, H., Natsume, M., Abe, H., 1999. Detection of antifungal activity in Anemarrhena asphodeloides by sensitive BCT method and isolation of its active compound. J Agric Food Chem 47, 584-587.

[7] Li, T. J., Qiu, Y., Yang, P. Y., Rui, Y. C., Chen, W. S., 2007. Timosaponin B-II improves memory and learning dysfunction induced by cerebral ischemia in rats. Neurosci Lett 421, 147-151.

[8] Ma, B. P., Wang, B. G., Dong, J. X., Yan, X. Z., Zhang, H. J., Tu, A. P., 1997. New spirostanol glycosides from Anemarrhena asphodeloides. Planta Medica 63, 376-379.

[9] Miura, T., Ichiki, H., Hashimoto, I., Iwamoto, N., Kato, M., Kubo, M., Ishihara, E., Komatsu, Y., Okada, M., Ishida, T., Tanigawa, K., 2001a. Antidiabetic activity of a xanthone compound, mangiferin. Phytomedicine 8, 85-87.

[10] Miura, T., Ichiki, H., Iwamoto, N., Kato, M., Kubo, M., Sasaki, H., Okada, M., Ishida, T., Seino, Y., Tanigawa, K., 2001b. Antidiabetic activity of the rhizoma of Anemarrhena asphodeloides and active components, mangiferin and its glucoside. Biol Pharm Bull FIELD Full Journal Title: Biological & pharmaceutical bulletin 24, 1009-1011.

[11] Oh, JK, Hyun, SY, Oh, HR, Jung, JW, Park, C., Lee, SY, Park, JH, Kim, SY, Kim, KH, Kim, YK, Ryu, JH, 2007. Effects of Anemarrhena asphodeloides on focal ischemic brain injury induced by middle cerebral artery occlusion in rats. Biological & pharmaceutical bulletin 30, 38-43.

[12] Ren, L. X., Luo, Y. F., Li, X., Wu, Y. L., 2007. Antidepressant activity of sarsasapogenin from Anemarrhena asphodeloides Bunge (Liliaceae). Pharmazie 62, 78-79.

[13] Takahashi, M., Konno, C., Hikino, H., 1985. Isolation and Hypoglycemie Activity of Anemarans A, B, C and D, Glycans of Anemarrhena asphodeloides Rhizomes1. Planta Med 51, 100-102.

The present inventors studied the pharmacological activity of the hair extract, and the effect of improving the memory disorder caused by cholinergic nervous system abnormalities caused by the hair extract and xanthone-based compounds manjiferin and neomanjiferrin and steroidal saponin thymosaponin AIII It was found that the present invention was completed.

Accordingly, it is an object of the present invention to provide a cholinergic nervous system damage improving agent comprising an extract of alcohol, water, or a mixed solvent of alcohol and water as an active ingredient.

Another object of the present invention is to provide a choline nervous system damage improving agent comprising fractions isolated from alcohol, water, or a mixed solvent extract of alcohol and water as an active ingredient.

Still another object of the present invention is a choline nervous system comprising at least one compound selected from the group consisting of mangiferin, neomangiferin, and timosaponin AIII, isolated and purified from hair extract. It is to provide a damage improver.

Another object of the present invention to provide a choline nervous system damage improving agent comprising norathyriol (norathyriol) derived from the hair extract as an active ingredient.

The present invention relates to choline nervous system damage improving agent containing alcohol, water, or a mixed solvent extract of alcohol and water as an active ingredient.

As used herein, alcohol refers to a straight or branched alcohol having 1 to 4 carbon atoms, and examples thereof include methanol, ethanol, butanol, and the like, but are not limited thereto.

The extract is prepared by drying it as it is or peeling it off, preferably using a mixed solvent of ethanol and water, most preferably 70% ethanol at room temperature or higher than 100 ° C. or lower.

On the other hand, the present invention relates to a cholinergic nervous system damage improving agent comprising as an active ingredient fractions isolated from alcohol, water, or a mixed solvent extract of alcohol and water. The manufacturing process of the fractions will be described in detail with reference to FIG. 1 as follows:

(i) extracting Jimo with alcohol, water, or a mixed solvent of alcohol and water, most preferably 70% ethanol and concentrating under reduced pressure;

(ii) resuspending the resulting extract in water and fractionating with methylene chloride to obtain a water fraction;

(iii) fractionating the obtained water fraction with butanol to obtain a butanol fraction;

(iv) concentrating the obtained butanol fraction under reduced pressure and inducing recrystallization in methanol, and washing the resulting precipitate with methanol to obtain a methanol insoluble fraction and a methanol soluble fraction; And

(v) column chromatography of the obtained methanol soluble fraction to obtain a fraction containing thymosaponin AIII as a main component, (iii) a butanol fraction, and (iv) a xanthone compound mannjiferin; A methanol insoluble fraction containing neomanziferin as a main component and a methanol soluble fraction containing a steroidal saponin as a main component can be prepared, respectively, in step (v), a thymosaponin AIII-containing fraction.

In step (v), column chromatography is preferably carried out on a silica gel column using a mixed solvent of methylene chloride, methanol and water. The mixed solvent is preferably methylene chloride: methanol: water 7: 1: 0.5-7: 3: 1.

On the other hand, the present invention relates to a cholinergic nervous system damage improving agent comprising at least one compound selected from the group consisting of manjiferin, neomanjiferrin and thymosaponin AIII separated and purified from the hair extract. The manjiperin and neomanjiferrin are methanol insoluble fractions obtained in step (iv), and thymosaponin AIII is conventional separation and purification methods, such as column chromatography, from the thymosaponin AIII-containing fractions obtained in step (v). It can manufacture by.

On the other hand, the present invention relates to a choline nervous system damage improving agent comprising a noratiriol as an active ingredient, the noratiriol is methanol insoluble containing the main component of mannferin and neomanjiferrin obtained in the above step (iv) Fractions can be prepared by treating human intestinal bacteria.

[Formula 1]

Choline nervous system damage improving agent of the present invention can be used as a pharmaceutical composition for the treatment or prevention of memory loss and brain damage caused by choline nervous system damage.

The pharmaceutical composition of the present invention may be administered orally (dose or inhalation) or parenterally (eg, intravenous, subcutaneous, transdermal, rectal), and may be used in tablets, capsules, depending on the purpose of use. , Granules, fine subtilae, powders, sublingual tablets, suppositories, ointments, injections, emulsions, suspensions, drug-treated syrups and the like. The various types of drugs include excipients, binders, disintegrators, lubricants, preservatives, antioxidants, isotonic agents, buffers, coatings, sweeteners, solubilizers, bases, dispersants, stabilizers, coloring agents, and the like. It is prepared by known techniques using pharmaceutically acceptable carriers conventionally used in the form of medicaments.

The content of the extract or compound of the present invention in the preparation of the medicament depends on the form of the medicament, but is at a concentration of about 0.01 to 100% by weight.

The dosage of the pharmaceutical composition of the present invention varies in a wide range depending on the type of mammal including the person to be treated, the extent of the disease and the judgment of the physician. In general, however, in the case of oral administration, 0.01 to 500 mg of the active ingredient may be administered per day per 1 kg of body weight, and in the case of parenteral administration, 0.01 to 100 mg of the active ingredient may be administered per day of 1 kg of body weight. The daily dosages described above can be used one at a time or dividedly, and can be arbitrarily changed depending on the extent of the disease and the judgment of the physician.

On the other hand, the cholinergic nervous system damage improving agent of the present invention can also be used as a dietary supplement for the treatment or prevention of memory loss. There are no particular restrictions on the types of dietary supplements, and they are in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, or general foods such as candy, sweets, gum, ice cream, noodles, bread, and beverages. Can be added to.

The health functional food of the present invention may be prepared by appropriately using fillers, extenders, binders, wetting agents, disintegrants, sweeteners, fragrances, preservatives, surfactants, lubricants, excipients and the like in a conventional manner.

 The content of the extract or compound of the present invention in the manufacture of the health functional food depends on the form of the health functional food, but the concentration is about 0.01 to 100% by weight. Since the health functional food of the present invention has little toxicity and side effects, it can be used with confidence even for long-term use for the purpose of prevention.

The extract of the present invention and the compounds isolated therefrom, manjiferin, neomanjiferrin, thymosaponin AIII, and norathiolol exhibit excellent acetylcholinesterase inhibitory activity and memory loss and brain damage relieving effect by cholinergic nervous system damage. Therefore, the hair extract of the present invention and the compound isolated therefrom can be used as a pharmaceutical composition or health functional food for the prevention or treatment of memory loss.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. These Examples and Experimental Examples are only for illustrating the present invention, it is apparent to those skilled in the art that the scope of the present invention is not limited to these Examples and Experimental Examples.

In the following Examples and Experimental Examples, NMR spectroscopy was performed using Bruker AM-500 (500 MHz), TLC was used with Merck silica gel 60F ₂₅₄ , and the reagent was Sigma, Tacrine (9-amino-1,2,3,4-tetrahydroacridine hydrochloride) and (-) scopolamine hydrobromide purchased from USA were used, and all other reagents used ultrapure premium reagents. It was.

Example  1: Preparation of 70% Ethanol Extract of Hair

The hair was dried as it was or peeled, and then extracted in a water bath using 70% ethanol, followed by complete drying to prepare 70% ethanol extract of hair.

Example  2: Isolation of Active Ingredients from 70% Ethanol Extracts of Gemini

The 70% ethanol extract of the hairs obtained in Example 1 was resuspended in water, fractionated with CH ₂ Cl ₂ , the water layer was extracted with BuOH, and concentrated under reduced pressure. The concentrated BuOH extract was then recrystallized with MeOH and the crystallized precipitated precipitate was washed with MeOH to give methanol insoluble fraction and methanol soluble fraction. The methanol insoluble fraction was then subjected to silica gel column chromatography (top layer of chloroform: methanol: water = 65: 35: 10) to separate manjiperin and neomanziferin. On the other hand, the methanol soluble fraction was prepared by silica gel column chromatography using a mixed solvent of methylene chloride, methanol and water (CH ₂ Cl ₂ : MeOH: H ₂ O = 7: 1: 0.5-> 7: 2: 0.5-> 7: 3: 1) to separate thymosaponin AIII, thymosaponin BII, thymosaponin BIII, and thymosaponin D.

Timothy saponin AIII: white amorphous powder. FAB-MS m / z : 763.5 [M + Na] ⁺

¹³ C NMR (125 MHz) 30.971 (C-1). 27.031 (C-2), 75.522 (C-3), 30.971 (C-4), 36.963 (C-5), 26.833 (C-6), 26.833 (C-7), 35.557 (C-8), 40.260 (C-9), 35.278 (C-10), 21.174 (C-11), 40.341 (C-12), 42.503 (C-13), 56.483 (C-14), 32.183 (C-15), 81.376 ( C-16), 62.216 (C-17), 16.622 (C-18), 24.040 (C-19), 40.907 (C-20), 14.941 (C-21), 109.7 (C-22), 26.434 (C -23), 26.229 (C-24), 27.573 (C-25), 69.882 (C-26), 16.308 (C-27), 102.588 (3-Gal C-1 '), 81.879 (C-2') , 75.263 (C-3 '), 71.753 (C-4'), 76.650 (C-5 '), 62.824 (C-6'), 106.167 (O-3 Glu C-1 ”), 76.987 (C-2 ”), 78.459 (C-3”), 72.987 (C-4 ”), 78.064 (C-5”), 65.129 (C-6 ”).

Manziferin: light green amorphous powder.

¹³ C NMR (100 MHz) 162.254 (C-1), 108.04 (C-2), 164.295 (C-3), 93.813 (C-4), 103.088 (C-5), 154.606 (C-6), 144.228 (C-7), 108.489 (C-8), 179.551 (C-9), 156.697 (C-4a), 151.286 (C-4b), 112.128 (C-8a), 101.772 (C-8b), 82.025 ( 2-glc C-1 '), 73.564 (C-2'), 71.103 (C-3 '), 70.724 (C-4'), 79.449 (C-5 '), 61.961 (C-6').

Neomanziferin

¹³ C NMR (100 MHz) 162.5 (C-1), 108.3 (C-2), 164. 5 (C-3), 94.0 (C-4), 103.3 (C-5), 156.9 (C-6) , 144.4 (C-7), 112.4 (C-8), 179.8 (C-9), 154.7 (C-4a), 151.5 (C-4b), 108.8 (C-8a), 102.0 (C-8b), 73.8 (2-glc C-1 '), 71.3 (C-2'), 79.7 (C-3 '), 71.0 (C-4'), 82.2 (C-5 '), 61.4 (C-6') , 103.4 (7-glc C-1``) 73.5 (C-2 ''), 76.1 (C-3``), 69.6 (C-4 ''), 77.3 (C-5 ''), 60.7 ( C-6 '').

Example  3: from methanol insoluble fraction Norathiolol  Produce

The methanol insoluble fraction (100 mg) obtained in Example 2 was sterilized in TS broth medium (100 ml) and suspended in a fecal suspension (1 gram of fecal matter in 20 ml of TS medium) and left for 1 hour in a healthy Korean. The supernatant was taken and centrifuged at 10,000 rpm for 10 minutes, and the precipitate was added to the same suspension in 10 ml of the same medium) and cultured for 24-72 hours. After extracting twice with 100 ml of ethyl acetate and concentrating, the concentrate (90 mg) was purified by silica gel column chromatography (3 × 20 cm; chloroform: MeOH = 10: 1) to separate 21 mg of noratiriol.

Norathithiol: yellow crystals. EIMS 260.049 [M] ⁺

UV λ max (EtOH) nm 238 (3.98), 256 (3.99), 311 (3.93), 362 (3.86).

¹³ C NMR (100 MHz): 162.7 (C-1), 97.7 (C-2), 164.7 (C-3), 93.6 (C-4), 102.7 (C-5), 154.1 (C-6), 143.8 (C-7), 108.1 (C-8), 178.9 (C-9), 157.3 (C-4a), 151.0 (C-4b), 111.8 (C-8a), 101.6 (C-8b).

Experimental Example  One: Acetylcholinesterase  Inhibition activity measurement

Acetylcholinesterase (AChE) inhibitory activity was measured using a microplate assay modified from the Elllman method. 125 μl of 3 mM DTNB (5,5'-dithiobis- [2-nitrobenzoic acid]), 25 μl of 15 mM acetylthiocholine, 25 μl of test substance dissolved in buffer, and 50 μl of buffer were prepared in 96 wells. Was added to each. Then, a microplate reader (uQuant MQX200, BioTek) was measured 10 times at 40 seconds intervals at 405 nm. Thereafter, enzyme (25 μl of AChE at 0.226 U / ml) was added and measured in the same manner. Then, the reaction rate before and after the addition of the enzyme was calculated, and showed the acetylcholinesterase inhibitory activity as a percentage compared to the group without addition of the test substance.

Table 1: Acetylcholinesterase Inhibitory Activity

group density Inhibition Rate (%) Vehicle alone - - Jimo's 70% Ethanol Extract 1 mg / ml 14.17 ± 3.11 10 mg / ml 57.11 ± 2.51 Methanol Insoluble Fractions (Manjiferin and Neomanziferin Containing Fractions) 0.1 mg / ml 23.22 ± 1.49 1 mg / ml 79.92 ± 2.99 Fraction containing thymosaponin AIII 0.1 mg / ml 18.12 ± 1.81 1 mg / ml 60.19 ± 3.18 Manziferin 0.01 mM 14.86 ± 3.18 0.1 mM 76.89 ± 1.40 Neomanziferin 0.01 mM 7.51 ± 1.19 0.1 mM 55.19 ± 3.21 Noratiriol 0.01 mM 25.21 ± 2.19 0.1 mM 80.11 ± 2.31 Timothy saponin AIII 0.01 mM 9.07 ± 4.19 0.1 mM 57.11 ± 2.44

Each value is mean ± S.E.M (n = 5)

Experimental Example  2: choline nervous system damage In mice  Measures to improve memory

All animals used in the experiment were performed with 28-30 g of male Institute of Cancer Research (ICR) -based mice according to the Guidelines for Use and Care of Laboratory Animals (NIH publication No. 85-23). Experimental animals were housed 5-6 per cage and left free to eat. The temperature in the kennel was 23 ± 1 ° C, the humidity was 60 ± 10%, and 12 hours (07:30 ~ 19:30) was maintained at all times.

Experimental Example  2-1: Activity Measurement Using Passive Avoidance Assay

The test group was orally administered after dissolving the test substance in 10% Tween 80 (5 ml / kg), the control group was orally administered 10% Tween 80 at the same dose. 30 minutes after oral administration, 1 mg / kg of scopolamine, a cholinergic nervous system drug dissolved in distilled water, was intraperitoneally administered, and 30 minutes after scopolamine administration, the mouse was placed in a brightly lit compartment (50 W bulb). After 10 seconds of placement, the guillotin door (5x5 cm) was opened to enter the dark compartment (Gemini Avoidance System; San Diego, USA). Each compartment was 20x20x20 cm of space. The time from the opening of the guillotine door to the dark side of the mouse was measured (acquisition trial). Once the mouse has gone four rounds into the dark, the guillotine door closes, and an electric shock of 0.5 mA flows through the bottom of the grid for three seconds, which the mouse remembers.

Passive avoidance experiments were conducted 24 hours after the acquisition test to determine the effect of the test substance. After 10 seconds of search time, the time required to open the guillotine door and enter the four feet of the mouse toward the dark (latency time) was measured up to 300 seconds (retention trial). The longer it takes to go to the dark, the better the memory.

Table 2: Activity measurement results by passive avoidance method

group Acquisition test score (mean ± SEM) Memory test score (mean ± SEM) Vehicle alone 23.6 ± 2.2 271.5 ± 13.6 Spopolamine Sole 31.8 ± 6.9 29.4 ± 7.0 70% Ethanol Extract (50mg / kg) and Scopolamine from Hemo 29.5 ± 5.9 65 ± 29.0 Methanol Insoluble Fraction (Manjiferin and Neomanziferin Containing Fraction) (50mg / kg) and Scopolamine 26.9 ± 5.0 125.4 ± 26.2 Fraction containing thymosaponin AIII (50 mg / kg) and scopolamine 26.2 ± 4.7 99.1 ± 29.0 Manziferin (20mg / kg) and Scopolamine 23.7 ± 5.0 160.4 ± 26.2 Neomanziferin (20mg / kg) and scopolamine 24.5 ± 3.9 71.1 ± 22.1 Timothy saponin AIII (20 mg / kg) and scopolamine 27.4 ± 5.4 73.2 ± 16.26 Norathiolol (20mg / kg) and Scopolamine 26.1 ± 4.1 170.8 ± 20.9

Experimental Example  2-2: Morris Water Maze  Activity Measurement Using Experimental Methods

The Morris water maze test was performed in a tank containing 500 ml of milk at 20 ± 1 ° C, 30 cm deep in a circular bath 90 cm in diameter and 45 cm high at low light. The tank was divided into four virtual zones, one of which was placed at a height of 29 cm so that a 6 cm diameter scaffold was submerged about 1 cm. On the first day of training, the swim training was performed without a scaffold for 60 seconds, and the rest of the training was performed four times daily to find the scaffold (training trial). He was allowed to stay for 10 seconds on the scaffold and rest on the scaffold for 10 seconds if no scaffold was found for 60 seconds. After the training, the experimental animals were dried with an ultraviolet lamp. The training was repeated every 30 seconds, and the time required to find the scaffold was measured with a video camera. In the final experiment, after removing the scaffold from the tank, the experimental animals were placed in the tank and the time spent on the scaffold was measured (probe trial). The experiment was performed 1 hour after oral administration of tacrine (10 mg / kg) as a positive control drug. Memory impairment was induced by intraperitoneal administration of scopolamine at the 0.8 mg / kg level, and the control group was given only 10% Tween 80.

Table 3: Activity measurement results by probe test

group Swimming time (average ± SEM) Vehicle 25.4 ± 1.4 Scopolamine 14.8 ± 0.7 Manziferin (20mg / kg) and Scopolamine 20.3 ± 1.4 Neomanziferin (20mg / kg) and scopolamine 17.3 ± 1.2 Timothy saponin AIII (20 mg / kg) and scopolamine 18.1 ± 1.0 Norathiolol (20mg / kg) and Scopolamine 22.3 ± 1.3

Table 4: Results of activity measurements by training test

group Swimming time (average ± SEM) 1 day 2 days 3 days 4 days Vehicle 44.3 ± 3.5 19.0 ± 3.1 12.2 ± 2.2 7.5 ± 1.7 Scopolamine 54.9 ± 2.4 44.9 ± 2.7 37.5 ± 3.0 33.2 ± 3.2 Manziferin (20mg / kg) and Scopolamine 40.1 ± 3.4 30.6 ± 3.7 26.9 ± 3.6 20.5 ± 3.0 Neomanziferin (20mg / kg) and scopolamine 43.9 ± 3.6 35.1 ± 3.6 31.1 ± 3.2 26.1 ± 3.3 Timothy saponin AIII (20 mg / kg) and scopolamine 41.9 ± 3.5 34.9 ± 3.4 26.5 ± 3.0 19.8 ± 2.9 Norathiolol (20mg / kg) and Scopolamine 37.3 ± 2.7 27.4 ± 2.5 22.8 ± 3.4 16.7 ± 3.1 Taclean 47.7 ± 3.3 34.0 ± 3.8 24.1 ± 3.4 16.2 ± 2.9

1 is a view showing a process for separating the active ingredient from the 70% ethanol extract of the hair.

Claims (10)

Choline nervous system damage improving agent comprising as an active ingredient at least one compound selected from the group consisting of mangiferin, neomangiferin (neomangiferin) and timosaponin AIII (timosaponin AIII). 2. The cholinergic nervous system damage improving agent according to claim 1, comprising manziferrin, neomanjiferrin, or a mixture of manjiferin and neomanjiferrin as an active ingredient. Choline nervous system damage improver comprising norathyriol as an active ingredient. Choline nervous system damage improving agent comprising alcohol, water, or a mixed solvent extract of alcohol and water as an active ingredient. 5. The choline nervous system damage improving agent according to claim 4, wherein the extract is 70% ethanol extract. (i) extracting gimo with alcohol, water, or a mixed solvent of alcohol and water and concentrating under reduced pressure; (ii) resuspending the resulting extract in water and fractionating with methylene chloride to obtain a water fraction; And (iii) a cholinergic nervous system damage improving agent comprising the butanol fraction of the hair prepared as an active ingredient, comprising the step of preparing the butanol fraction by fractionating the obtained water fraction with butanol. (i) extracting gimo with alcohol, water, or a mixed solvent of alcohol and water and concentrating under reduced pressure; (ii) resuspending the resulting extract in water and fractionating with methylene chloride to obtain a water fraction; (iii) fractionating the obtained water fraction with butanol to obtain a butanol fraction; And (iv) concentrating the obtained butanol fraction under reduced pressure and inducing recrystallization in methanol, and then washing the resulting precipitate with methanol to obtain a methanol insoluble fraction. Cholinergic damage modifier. 8. The cholinergic nervous system damage improving agent according to claim 7, wherein the methanol insoluble fraction of the hairs contains manjiperin and neomanjiperin as main components. (i) extracting gimo with alcohol, water, or a mixed solvent of alcohol and water and concentrating under reduced pressure; (ii) resuspending the resulting extract in water and fractionating with methylene chloride to obtain a water fraction; (iii) fractionating the obtained water fraction with butanol to obtain a butanol fraction; (iv) concentrating the obtained butanol fraction under reduced pressure and inducing recrystallization in methanol, then washing the resulting precipitate with methanol to obtain a methanol soluble fraction; And (v) Cholinergic nervous system damage improving agent comprising thymosaponin AIII-containing fractions prepared as an active ingredient, comprising the step of column chromatography of the obtained methanol soluble fractions to obtain a fraction containing thymosaponin AIII as a main component. 10. The choline nervous system damage improving agent according to any one of claims 1 to 9, wherein the choline nervous system damage improving agent is used for the treatment or prevention of memory loss and brain damage caused by cholinergic border damage.

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