KR102125031B1 - Composition for Improving or Enhancing Memory Comprising Calystegia soldanella Extracts - Google Patents

Abstract

The present invention is Calystegia soldanella ) A composition for improving and enhancing memory, including extract, Calystegia soldanella ) It relates to a composition for improving memory and learning ability comprising an extract. According to the present invention, learning ability and taste reminiscent memory can be increased through ingestion of Calystegia soldanella extract, which is one of the salt plants, and can be usefully used as a candidate for new drugs and health functional materials.

Description

Composition for Improving and Enhancing Memory Containing Seaweed Extract Extract {Composition for Improving or Enhancing Memory Comprising Calystegia soldanella Extracts}

The present invention is Calystegia soldanella ) A composition for improving and enhancing memory, including extract, Calystegia soldanella ) It relates to a composition for improving memory and learning ability comprising an extract.

Chlorine vegetation is a plant that grows on salty soils, and because it grows on salty soils, the reservoir structure that stores a lot of water in cells is greatly developed. Salicornia , a kind of salt plant herbacea L.) has been shown to be effective against adult diseases such as hypertension, diabetes and hyperlipidemia (Bang MA, et. al . 2002. Hypoglycemic and anti-oxidant effect of dietary hamcho powder in streptozotocin-induced diabetic rats. J Korean Soc Food Sci Nutr 31:840-846.), commercialized as a functional food material. Suaeda asparagoides contains essential nutrients such as proteins, fats, and trace elements such as minerals, phosphorus, calcium, iron, sodium, and mitamines A, B ₁ , B ₂ , C, etc. Has also been used as (Lee BH, et al . (2002). Growth characteristics and it's potentiality of use of halophyte, Suaeda asparagoides Miq. Korean J Int Agri 14(2):87-93.). Especially calystegia soldanella (Calystegia soldanella) is to medicinal roots and young shoots, roots, and as hyoseon chogeun (孝扇草根), known to be effective in pain relief, diuretic, oxygen species (Huang, Z., & Feng, C. (2010) .Experimental study on anti-inflammatory and analgesic effects of water extracts of Calystegia soldanella . Chinese Archives of Traditional Chinese Medicine , 28(6), 1142-1143.).

Parkinson's disease (Parkinson's disease) is one of the neurodegenerative diseases, leading to the destruction of dopaminergic neurons, causing dementia, and disrupting mitochondrial function. Also, there are cases in which peripheral sensations such as taste, smell, pain, and vision are lower than those of the general public. Drosophila , an animal model melanogaster ) exists in Parkinson's disease model Drosophila (DJ-1β ^ex54 ). The physiological functions of DJ-1 are presumed to be antioxidant proteins, stimulators of oxidative stimuli, or proteolytic enzymes (Zang, LM, & Shimoji. (2005).Mitochondrial location of the Parkinson's disease related protein DJ-1 implications for pathogenesis. Hum Mol Genet , 14 , 2063-2073.). DJ-1β ^ex54 (hereinafter referred to as DJ-1), which lacks the DJ-1 gene, significantly reduces the mitochondrial function of synthesizing ATP when subjected to oxidative stress such as aging.

Taste associative memory assay has been established as an experimental method to confirm learning ability and taste-associated memory (Poudel, S., & Lee, Y. (2018).Improved Taste Associative Memory and Memory Enhancement by Feeding Omija in Parkinson's Disease Fly Model. Molecules and cells .). This test method, like the Pavlovian theory, discriminates learning ability and taste association memory by whether or not it can act conditionally. Caffeine, a chemical avoided by fruit flies, and sucrose, a chemical preferred by fruit flies, are used. In Drosophila, neutrophil sucrose and unconditional stimulus caffeine were linked to generate a conditional response that did not extend to sucrose, a conditional stimulus that was irrelevant to not taking action to reach for food. Using these experiments, it has been found that intake of Omija improves the memory associated with the taste of Drosophila.

Under these technical backgrounds, the present inventors searched for a salty plant extract that improves memory when ingested in Drosophila, and as a result of diligent efforts to develop new drugs and health functional materials, it was confirmed that intake of snail flower extract improves memory associated with taste in yellow fruit flies. And completed the present invention.

The above information described in the background section is only for improving the understanding of the background of the present invention, and therefore does not include information forming prior art already known to those of ordinary skill in the art. It may not.

An object of the present invention is to provide a composition for improving and improving memory, including a salt plant extract that improves memory when ingested.

Another object of the present invention is to provide a food composition for improving memory or learning ability, including a salt plant extract that improves memory when ingested.

In order to achieve the above object, the present invention is Calystegia soldanella ) provides a composition for improving and improving memory.

The present invention also, Calystegia soldanella ) It provides a food composition for improving memory or learning ability containing the extract.

According to the present invention, one of the salt plants, Calystegia soldanella ) It is possible to increase learning ability and memory associated with taste through intake of extract, and it can be usefully used as a new drug, a candidate for health functional material, etc.

1 is a result of comparing and comparing the learning ability and taste association memory of wild type Drosophila and DJ-1β ^ex54 Drosophila (repeat 7 times).
Figure 2 is a result of measuring the learning ability and taste reminiscent memory of Drosophila that ingested the oleander extract (repeat 6-7 times), Figure 2A is a wild type fruit fly intake of the oleander extract, Figure 2B intake It is a result of measuring the learning ability and memory of taste in a DJ-1β ^ex54 Drosophila. The asterisk means that a significant change occurred when compared to the control feeding group, and the statistical significance is a single factor ANOVA with Scheffe's analysis as a post hoc test to compare two data sets (*P<0.05, **P<0.01). It was derived by analysis, and the error bar means SEMs.
Figure 3 is a result of measuring the learning ability and taste association memory of fruit flies ingested Namunjae extract (repeat 6-7 times).
4 is a result of measuring the learning ability and taste reminiscent memory of Drosophila ingested Cheonil Sedge extract (repeat 6-7 times).
Figure 5 is a result of measuring the learning ability and taste association memory of Drosophila ingested Nammija extract (repeat 6-7 times).
6 is a result of measuring the learning ability and taste reminiscent memory of Drosophila ingesting the extract of Cheonmun-dong (repeat 6-7 times)
7 is a result of measuring the learning ability and memory associated with taste of fruit flies ingested seaweed extract (6-7 repetitions).
Figure 8 is a result of measuring the learning ability and taste association memory of fruit flies ingested the extract of the marsupial (6-7 repetitions).
Figure 9 is a result of measuring the learning ability and taste association memory of Drosophila ingested Gaetganghwa extract (6-7 repetitions).
10 is a result of measuring the learning ability and taste reminiscent memory of Drosophila that ingested the extract of the ants (6-7 repetitions).
11 is a result of measuring the learning ability and memory associated with taste of Drosophila ingesting the extract of oleander (6-7 repetitions).
12 is a result of measuring the learning ability and memory associated with taste of Drosophila ingesting the magpie swimming extract (repeat 6-7 times).
13 is a result of measuring the learning ability and taste reminiscent memory of Drosophila that ingested the extract of Termites (6-7 repetitions).

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by a person skilled in the art to which the present invention pertains. In general, the nomenclature used herein is well known in the art and is commonly used.

In an embodiment of the present invention, when fruit flies ingested food containing a salt plant extract, it was confirmed whether it affects the learning ability and taste association memory associated with taste memory, and in particular, in fruit flies that consumed the extract of oleander It was confirmed that the learning ability and memory were improved.

Therefore, the present invention is, in one aspect, Calystegia soldanella ) relates to a composition for improving and improving memory.

In the present invention, the sea buckthorn extract is water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane and 1,3-butylene It may be characterized by being extracted with one or more solvents selected from the group consisting of glycols, but is not limited thereto. Preferably, 99.9% methyl alcohol may be characterized as being extracted with a solvent.

In one embodiment of the present invention, in order to identify the salt plant extracts affecting learning ability and memory, Namunjae ( Suaeda asparagoides , lot number: PB2619.2), hairy pea ( Lathyrus japonica var . aleuticus , presale number: PB3492.2), Cheonil sedge ( Carex scabrifolia , presale number: PB1838.1), astronomical dong ( Asparagus cochinchinensis , Pre-sale number: PB2089.2), Haemul namul ( Suaeda maritima , Sale number: PB2620.3), Calystegia soldanella , Sale number: PB4285.4), Plantago major var . japonica for . yezomaritima , Sales number: PB4564.2), Gaetganghaeng ( Angelica japonica, pre-sale code: PB4043.2), Conger Michaelmas daisy (Aster tripolium , Presale No.: PB4826.1), Fortune Oil Herb ( Peucedanum japonicum , Presale No.: PB4051.2), Great Magpie Swimming ( Lysimachia mauritiana , Presale No.: PB4143.1), Forsythia ( Spergularia marina , Pre-sale number: PB2646.1), Salicornia herbacea, pre-sale code: PB2617.1) and namohmija (Kadsura japonica, pre-sale code: accept pre-sale at PB2896.4) to extract plant extracts Korea Bank (Korea Plant Extract Bank) was found in Drosophila learning ability and taste reminiscent of memory.

In the present invention, the composition may be characterized by enhancing taste-associated memory.

In the present invention, it was confirmed that the Gaetme flower extract has an effect of improving learning ability and an excellent effect of improving memory, and may be included in a pharmaceutical composition for improving and enhancing memory.

The pharmaceutical composition comprising the extract of saffron according to the present invention may further include suitable carriers, excipients and diluents commonly used in the manufacture of ills.

The pharmaceutical dosage form of the extracts according to the present invention can also be used in the form of their pharmaceutically acceptable salts, and can also be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable set.

The pharmaceutical composition comprising the extract according to the present invention is in the form of an oral dosage form such as powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, external preparation, suppository and sterile injectable solution, respectively, according to a conventional method. It can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the extract 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. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ) Or by mixing lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate and talc are used in addition to simple excipients. Liquid preparations for oral use include suspensions, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be included. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

In another aspect, the present invention, Calystegia soldanella ) It relates to a food composition for improving memory or learning ability comprising an extract.

The food composition according to the present invention may be added to the health functional foods such as food and beverages for the purpose of improving memory or improving learning ability.

In the present invention, there is no particular limitation on the type of food. Examples of foods to which the above substances can be added are drinks, meat, sausage, bread, biscuits, rice cake, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, Beverages, alcoholic beverages and vitamin complexes, dairy products and dairy products, etc., and include all functional foods in the normal sense.

Gaetme flower extract of the present invention may be added as it is to foods or used with other foods or food ingredients, and may be suitably used according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to its purpose of use (for prevention or improvement). In general, the amount of the Gaetme flower extract in the dietary supplement can be added at 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for health and hygiene purposes or for health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

In the present invention, the health functional beverage composition is an essential component in the indicated proportions, and there is no particular limitation on other components except for containing the Gaetme flower extract, and various flavors or natural carbohydrates, etc., as additional components, such as ordinary beverages It can contain. Examples of the natural carbohydrates described above include monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, etc.; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatine, stevia extract (for example, rebaudioside A, glycyrrhizine, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 1 to 20 g per 100 of the composition of the present invention, preferably about 5 to 12 g.

In addition to the above, the extract of the seaweed flowers of the present invention is a variety of nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid And salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid used in carbonated beverages, and the like.

In addition, the extract of Gaetme flower of the present invention may contain natural fruit juice and fruit juice for the production of fruit juice beverages and vegetable beverages.

These ingredients can be used independently or in combination. The proportion of these additives is not so critical, but it is generally selected from the range of 0.1 to about 20 parts by weight per 100 parts by weight of the seaweed extract of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

<Experiment Method>

The method used in this experiment is as follows.

Fruit fly Stark ( stock )

DJ-1β ^ex54 was used as a Parkinson's disease model fruit fly in Bloomington Stock Centor, and w ¹¹¹⁸ was used as a wild type control.

compound

Sucrose (Cucine, CAS. No.: 57-50-1) and caffeine (Caffeine, CAS. No.: 58-08-2) are from Sigma-Aldrich Company (St. Louis, MO, USA). Purchased.

Chlorine plant extract: Suaeda asparagoides , lot number: PB2619.2), hairy pea ( Lathyrus japonica var . aleuticus , presale number: PB3492.2), Cheonil sedge ( Carex scabrifolia , presale number: PB1838.1), astronomical dong ( Asparagus cochinchinensis , Pre-sale number: PB2089.2), Haemul namul ( Suaeda maritima , Sale number: PB2620.3), Calystegia soldanella , Sale number: PB4285.4), Plantago major var . japonica for . yezomaritima , Sales number: PB4564.2), Gaetganghaeng ( Angelica japonica, pre-sale code: PB4043.2), Conger Michaelmas daisy (Aster tripolium , Presale No.: PB4826.1), Fortune Oil Herb ( Peucedanum japonicum , Presale No.: PB4051.2), Great Magpie Swimming ( Lysimachia mauritiana , Presale No.: PB4143.1), Forsythia ( Spergularia marina , Pre-sale number: PB2646.1), Salicornia herbacea, pre-sale code: PB2617.1), and namohmija (Kadsura japonica, pre-sale code: PB2896.4) extract has received pre-sale at the Bank of Korea plant extracts (Korea Plant Extract Bank). The salt plant extract was diluted to 0.1% and dissolved in Drosophila medium for use.

Learning & Memory Test ( Learning & Memory test )

The learning & memory test was conducted in a similar way by modifying what is known in the existing literature. More specifically, Drosophila, which is 2-4 days old, were first cultured for 7-8 days in a control medium or a medicinal plant extract medium. Fifteen to 18 hours before the test, Drosophila folds the Kimtech wiper to a size that can be placed in a vial, puts it in an empty vial, and soaks the wiper with drinking water rather than distilled water. Store each fruit fly in a vial with a wiper. On the day of the test, the vial with fruit flies is stunned by putting it on ice, and then the fruit flies are placed on a slide glass and the wings are fixed with nail polish. Store it in a dry place for about an hour to recover from fainting. Moisturize the fruit flies by soaking them in the wiper until they are no longer drinking. Thereafter, a wiper moistened with a 500 mM sucrose solution is placed on the leg of the fruit fly, and only the fruit fly that extends the mouth (proboscis) is selected and tested. Conditional reflex training is conducted with 500 mM sucrose and 50 mM caffeine, and is repeated 15 times. During the test, Drosophila's thirst can take action to drink the sucrose solution and the caffeine solution. To prevent this behavior, after the 5th, 10th and 15th training, use a wiper moistened with water to replenish the fruit flies. When the wiper with 500mM sucrose is supplied to the Drosophila leg, the mouth is wiped with a 50mM caffeine-soaked wiper. However, it is necessary to prevent fruit flies from ingesting wipers soaked in sucrose aqueous solution and caffeine aqueous solution. After the conditional reflex training is completed, check whether or not the mouth is stretched by exposing sucrose aqueous solution to the leg every designated time (0, 5, 15, 30, 45, 60 minutes). We do not recommend running this experiment in a dry environment.

Example 1: Drosophila Mouth end extension reflex( Proboscis Extension Response ) analysis

When a normal Drosophila is starved for 15-18 hours and then exposed to 500mM sucrose aqueous solution on the legs, one poses to ingest the aqueous solution, one of which is the action of extruding the mouth (proboscis reflex, Proboscis Extension Response) ; PER). Conversely, when exposed to a compound that Drosophila dislikes, such as caffeine, it stops reaching out. Based on this behavior, a pretest related to taste memory was conducted. Wild-type fruit flies that have undergone controlled feeding have a PER of 100% in preliminary experiments and respond 100% to 500 mM sucrose (FIG. 1).

Example 2: Learning ability and taste association memory of wild type Drosophila and Parkinson's disease model Drosophila

Sucrose aqueous solution causes PER, but 50mM caffeine aqueous solution can suppress PER. In the early stage of training, Drosophila takes PER when exposed to 500 mM sucrose, but if PER is taken after sucrose exposure, caffeine aqueous solution is suppressed to suppress PER, and if sucrose exposure is PER, caffeine aqueous solution is exposed. This training is repeated 15 times, and as the training is repeated, the percentage of PER decreases. As the training continues, the PER ratio gradually decreases to about 12%, and it can be seen that the training is remembered (FIG. 1 ). There was no significant difference in the PER ratio between wild-type Drosophila and Parkinson's disease model Drosophila DJ-1β ^ex54 (Table 1).

PER(%) training Wild type DJ-1β ^ex54 Episode 5 45.5 38.5 Episode 10 25.0 23.6 Episode 15 12.3 12.7

After training, in this experiment, only sucrose is exposed, so the ratio of PER increases. The PER ratio gradually increases each time the experiment is continued, and it can be confirmed by digitizing the appearance of forgetting the memory of the training. The DJ-1 mutant showed a similar pattern to the wild type (Table 2).

PER(%) Experiment Wild type DJ-1β ^ex54 0 minutes 11.1 15.6 5 minutes 23.2 39.1 15 minutes 43.4 63.1 30 minutes 58.5 79.2 45 minutes 74.1 87.9 60 minutes 90.0 91.3

Compared to the wild type, 5 minutes (wild type: 23%, DJ-1β ^ex54 : 39%), 30 minutes (wild type: 58%, DJ-1β ^{ex54 :} 79%) and 45 minutes (wild type: 74%) of this ^experiment . DJ-1β ^ex54 : 87%) was found to have significantly increased the PER ratio of DJ-1β ^ex54 (Fig. 1). This means that the DJ-1 mutant, a Parkinson's disease model, does not differ significantly from the wild type in the ability to progress training, but has a significantly lower taste association memory.

Example 3: Confirmation of learning ability and taste association memory by intake of various types of salt plant extracts

In order to investigate the efficacy when ingesting the salt plant extract, the difference between the group cultured in the medium containing each salt plant and the culture medium in the control medium was examined for 7-8 days. The extracts of salty plants were extracted from Gaetme flower, Namunjae, Cheonil Sado, Namomija, Cheonmundong, Haehongnamul, Gaetul wonder, Gaetgang bow, Gaetgae taste, Gaetum oil herb, Gaetchi swimming, and Gaetamjari respectively.

The taste association memory test was conducted with Drosophila cultured in the control medium and the salt plant extract medium, and the taste association memory was improved in wild type Drosophila which ingested the buckthorn extract compared to other salt plant extracts (FIGS. 2 to 13). ).

There was no significant difference in the training ability of Drosophila cultured in the control medium and the medium containing the starfish extract. On the other hand, in this experiment, the taste association memory was significantly enhanced at 5 minutes, 30 minutes, 45 minutes and 60 minutes (Fig. 2A). It can be seen that, compared to Drosophila cultured in the control medium, the taste association memory of the Drosophila cultured in the medium containing the starfish extract is enhanced (Table 3).

PER(%) Experiment Control feeding Feeding 0 minutes 11.1 0.0 5 minutes 22.4 13.3 15 minutes 42.4 29.7 30 minutes 59.2 40.1 45 minutes 73.5 60.0 60 minutes 86.8 72.1

Similarly, Parkinson's disease model Drosophila conducted an experiment. When training, controlling eating a DJ-1β ^ex54 showed a PER ratio is not much different from eating calystegia soldanella DJ-1β ^ex54. However, in this experiment, the DJ-1β ^ex54 variant showed a significant difference from that ingesting the control medium. Taste association memory was significantly enhanced at 5, 15 and 30 minutes (Figure 2B). It can be seen that, compared to Drosophila cultured in the control medium, the taste reminiscent memory of Drosophila cultured in the medium containing the starfish extract was enhanced (Table 4).

PER(%) Experiment Control feeding Feeding 0 minutes 15.6 4.2 5 minutes 40.8 23.1 15 minutes 61.8 44.9 30 minutes 75.9 53.9 45 minutes 83.5 66.8 60 minutes 86.5 76.0

As a result, it was confirmed that when the wild type Drosophila and Parkinson's disease model Drosophila were fed with saffron extract, learning ability increased and memory was significantly improved.

Since the specific parts of the present invention have been described in detail above, it will be apparent to those of ordinary skill in the art that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby. will be. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (4)

A composition for improving memory or improving learning ability, which includes Calystegia soldanella extract.
The method of claim 1, wherein the extract is water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane and 1,3-butylene glycol Composition for improving memory or learning ability, characterized in that extracted with one or more solvents selected from the group consisting of.
The composition for improving memory or learning ability according to claim 1, wherein the composition enhances taste-associated memory.
Calystegia soldanella (Calystegia soldanella ) Food composition for improving memory or learning ability, including extracts.

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