KR20150090646A - Composition containing garlic sprouting extract for preventing or treating brain disease - Google Patents
Composition containing garlic sprouting extract for preventing or treating brain disease Download PDFInfo
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- KR20150090646A KR20150090646A KR1020140011542A KR20140011542A KR20150090646A KR 20150090646 A KR20150090646 A KR 20150090646A KR 1020140011542 A KR1020140011542 A KR 1020140011542A KR 20140011542 A KR20140011542 A KR 20140011542A KR 20150090646 A KR20150090646 A KR 20150090646A
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- garlic
- extract
- disease
- preventing
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- A—HUMAN NECESSITIES
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A61K36/8962—Allium, e.g. garden onion, leek, garlic or chives
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
Description
The present invention relates to a composition for preventing or treating brain diseases comprising garlic extract.
With the rapid increase in the elderly population, degenerative neurological diseases including brain, spinal, and peripheral nerve damage continue to increase. Degenerative nerve diseases include diseases related to nerve injury caused by ischemia and nerve injury caused by reactive oxygen species.
There are two mechanisms of neuronal death in cerebral ischemia, which are universally accepted. One is that when glutamate accumulates outside the cell by brain ischemia, the glutamate enters the cell and eventually causes excessive accumulation of intracellular calcium resulting in neuronal death (Kang TC, et al. In the ischemic-reperfusion period, a sudden supply of oxygen resulted in a decrease in the amount of N-methyl-D-aspartate receptors and excitatory amino acid carrier immunoreactivities in the CA1 area and subiculum after transient forebrain ischemia, J. Neurocytology, 30, pp945-955, In addition, there are two mechanisms by which oxidative neuronal cell death is caused by damage to DNA and cytoplasm due to the increase of in vivo radicals (Won MH, et al., Immunohistochemical detection of oxidative DNA damage induced by gerbil hippocampus in vivo , Brain Res., 836, pp 70-78, 1999; Sun AY, Chen YM, Oxidative stress and neurodegeneration disorders, J. Biomed. Sci., 5, pp 401-414, 1998).
In addition, damage to nerve cells is also caused by an increase in active oxygen in the blood, not by ischemia. The brain is shown to be more damaged by reactive oxygen species than any other organ in the body because 20% of the circulating O2 is concentrated in the brain, the cell membrane is rich in polyunsaturated fatty acid side chains, It has been reported in recent studies that it is associated with degenerative neurological diseases caused by induction of neuronal cell death because it is abundant in iron as a catalyst of radical reaction. For example, in the brain region of patients with Alzheimer's disease, there is an increase in the production of DNA damage, protein oxidation, lipid peroxidation, advanced glycosylation end products (AGE) ) Is increased to show the possibility of increased oxidative damage.
Currently, various treatments such as cell transplantation and surgical operation are proposed to treat these degenerative neurological diseases. However, most of them exhibit the risk factors and side effects, and the therapeutic agent which protects the damage of the nerve cell by virtue of the complexity of the damage mechanism is developed It is a fact that it does not become.
On the other hand, garlic (Allium savitum) is a plant distributed all over the world. It is also used for food, but it has been used for thousands of years by humans for various diseases such as hypertension, infection, wound, influenza and ulcer. In recent years, not only the anti-cancer effect and antimicrobial effect of garlic, but also the effect of garlic which lowers blood cholesterol and lowers the risk of cardiovascular disease, is attracting attention.
This garlic contains about 60% water, 28% carbohydrates (mainly fructan), 2.3% organic sulfur compounds, 2% protein (mainly allinase), 1.2% organic amino acids (mainly arginine), 1.5% It contains vitamins such as vitamin A, B1, B2, and C and calcium, potassium, copper, zinc, germanium, and other vitamins such as phytic acid (0.08%), saponin (0.07%), and sitosterol , Iron, magnesium, manganese, phosphorus, sulfur, selenium and the like.
The organosulfur compound is S-allyl-L-cystein sulfoxide, which contains alliin (1%), methine (0.12%), isoalignan (0.06% (0.6%) and glutamyl-S-allyl-L-cysteine in the L-glutamyl-S-allyl- And cysteine (0.4%).
Garlic contains alliin, which when garlic is scratched or crushed, changes to allicin by the action of an enzyme called alliinase, which is known as the main component of garlic It is very unstable at room temperature, so that the content of allysine is reduced to half in 2 to 16 hours and it is converted into another sulfur compound by the action of other enzymes, and it is known to cause a complex odor unique to garlic, It is characterized by very low solubility.
Thus, while extensive research into the various pharmacological effects of garlic and the composition of garlic has been carried out, the potential biological efficacy of garlic buds has hardly been studied compared to that of garlic. Germination of plant seeds usually promotes de novo synthesis of physiologically active compounds or phyroalexins. For this reason, the present inventors have completed the present invention by studying whether the germination of garlic increases the antioxidant activity as compared with that of raw garlic and produces a metabolite effective for physiological activity during germination.
It is an object of the present invention to provide a pharmaceutical composition for preventing or treating brain diseases containing an extract of garlic sprouts as an active ingredient.
It is still another object of the present invention to provide a food composition for preventing or ameliorating brain diseases containing garlic extract as an active ingredient.
In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating brain diseases containing garlic extract as an active ingredient.
The present invention also provides a food composition for preventing or ameliorating brain diseases, which contains garlic bud extract as an active ingredient.
The garlic extract of the present invention is superior to fresh germ which does not germinate the effect of protecting nerve cells, and thus can be useful for prevention or treatment of brain diseases.
FIG. 1 is a graph showing the results of FRAP assays of garlic sprout extract. FIG.
Fig. 2 shows DPPH radical scavenging activity of garlic extract.
FIG. 3 is a graph showing the results of evaluation of ABTS + radical scavenging activity of garlic extract.
4 is a graph showing glutamate-induced apoptosis inhibitory effect of garlic bud extract in HT22 cells.
FIG. 5 is a graph showing the effect of suppressing the production of glutamate-induced reactive oxygen species (ROS) by the extract of garlic sprouts in HT22 cells.
The present invention provides a composition for preventing or treating brain diseases containing garlic extract as an active ingredient.
The composition comprises a pharmaceutical composition and a food composition.
Hereinafter, the present invention will be described in detail.
The garlic extract of the present invention as an active ingredient of the present invention can be obtained by extracting by a conventional method. Typical extraction methods include, but are not limited to, ultrasonic extraction, filtration, and reflux extraction.
In the present invention, it was obtained by the following method.
First, the garlic sprouts are cleaned with water to remove foreign matter, and then dried and pulverized at room temperature. The ground garlic bud sample is extracted by immersion in a solvent. The extraction solvent is not limited thereto, but one or more solvents selected from water, alcohols having 1 to 4 carbon atoms, and mixed solvents thereof may be used, and ethanol is preferably used. After filtering the garlic bud extract, the solvent is evaporated and concentrated to give the final garlic bud extract.
The garlic bud extract of the present invention is excellent in nerve cell protection effect and can be useful for prevention or treatment of brain diseases.
Such brain diseases include degenerative brain diseases including but not limited to stroke, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Pick's disease, Creutzfeld-Jakob disease, and the like .
The composition of the present invention, together with the garlic extract, may further contain at least one known active ingredient having an effect of preventing or treating brain diseases.
The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral formulations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When the composition is formulated, it is prepared using a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant, which is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances and preservatives have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.
The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.
The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the individual, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. For the desired effect, the garlic extract of the present invention may be administered in an amount of 1 mg / kg to 1,000 mg / kg per day, and may be administered once or several times a day.
The pharmaceutical composition of the present invention may be administered in various ways in an individual. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.
The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention or treatment of brain diseases through nerve cell protection.
The composition of the present invention may be added to health functional foods for the purpose of prevention or treatment of brain diseases through nerve cell protection. When the garlic extract of the present invention is used as a food additive, the garlic extract may be used as it is or may be used together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, in the production of foods or beverages, Baechonggol extract of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.
There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, rice noodles, other noodles, gums, dairy products including ice cream, various soups, , An alcoholic beverage and a vitamin complex, and includes all the health foods in a conventional sense.
The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.
In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
Hereinafter, preferred embodiments and examples of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples, experimental examples and preparation examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples, experimental examples and preparation examples.
Example 1. Preparation of garlic extract and raw garlic extract
Garlic was a yellowish-green variety distributed from Namhae Garlic Research Institute (Namhae, Kyungnam, Korea). The garlic was peeled and germinated for 1 to 6 days in an incubator at 25 ° C, 100% humidity. Ungared fresh garlic and germinated garlic were minced and ground, and extracted with 10
Experimental Example 1. Confirmation of antioxidative effect of garlic extract
In order to confirm the antioxidant activity of the garlic extract, fresh germinated unsprozen and garlic shoots from 1 to 6 days of germination were extracted in the same manner as in Example 1, and the following experiment was conducted using the same as a sample.
1-1. FRAP Assay
In order to measure the antioxidant effect of the garlic extract of the present invention, when trivalent iron is lost to electrons and reduced to bivalent iron, TPRZ (ferric 2,4,6-tripyridyl-s-triazine) reagent reacts with bivalent iron The FRAP assay was performed using the principle of conversion to a blue material.
More specifically, 170 μL of distilled water, 7 μL of standard solution or sample, and 30 μL of FRAP solution were added to a 96-well plate and reacted at room temperature for 10 minutes. Absorbance was measured at 593 nm using an ELISA reader (Tecan Sunrise microplate reader, ReTiSoft Inc., Mississauga, Ontario, Canada) and trolox was used as a positive control. The results are shown in Fig.
As shown in FIG. 1, the garlic extract of the present invention exhibited a higher antioxidative activity in FRAP assay than the unextracted raw garlic extract. In particular, the garlic extract of the 5th day of germination had the best antioxidative activity Respectively.
1-2. Evaluation of DPPH radical scavenging ability
In order to confirm the antioxidative effect of the garlic extract of the present invention, the DPPH radical scavenging activity of the garlic extract was evaluated.
Specifically, 200 μl of a DPPH methanol solution was added to each well of a 96-well microplate, and 50 μl of each sample was added to each well. The plate was shaken briefly and the plate was incubated at 37 ° C. for 30 minutes Min. Absorbance was determined at 515 nm. DMSO and methanol were used as negative controls, and different amounts of alpha -tocopherol and trolox, respectively, were used as positive control. The results are shown in Fig.
As shown in Fig. 2, the garlic extract showed a DPPH radical scavenging ability that was much higher than that of fresh germinated garlic.
1-3. ABTS + Evaluation of radical scavenging ability
In order to confirm the antioxidative effect of the garlic extract of the present invention, the ABTS + method was performed.
Specifically, 5 ml of 7 mM ABTS solution and 2.45 mM potassium persulfate (80 μl) were mixed and incubated for 16 hours in an incubator in which light was blocked at room temperature to generate radicals. After diluting the radicals with ethanol at 734 nm to an absorbance of 0.7 ± 0.02, 90 μL ABTS solution and 10 μL sample were mixed and reacted at room temperature for 5 minutes and absorbance was measured at 734 nm. The ABTS + radical scavenging activity was calculated using the following equation. The results are shown in Fig.
As shown in FIG. 3, the garlic extract had an ABTS + radical scavenging activity, and in particular, the garlic extract of the 5th day of germination had the best antioxidant activity.
Experimental Example 2. Confirmation of protective effect of garlic bud extract on nerve cell
In order to confirm the neuronal cell protection activity of the garlic extract, fresh germinated germinated seeds and garlic shoots from 1 to 6 days of germination were extracted in the same manner as in Example 1, and the following experiment was conducted Respectively.
2-1. Cell preparation
Mouse hippocampal neuron HT22 cells were obtained from Dr. Song Kyung-sik (College of Pharmacy, Kyungpook National University) and maintained in DMEM medium containing 10% FBS, 100 units / ml penicillin and streptomycin at 37 ° C and 5% CO 2 .
2-2. Measurement of cell viability by MTT assay
In order to confirm the neuronal cell protection activity of garlic extract, MTT assay was performed to determine glutamate - induced HT22 cell viability.
The mouse hippocampal neuron HT22 cells prepared in 2-1. Above were cultured in a DMEM medium containing 10% FBS at a rate of 2 × 10 3 cells per well of a 96-well plate. On the next day, various concentrations of garlic bud extract and 5 mM glutamate were added to the cells. After 24 hours, the survival rate of the cells was measured by MTT assay. The results are shown in Fig.
As shown in FIG. 4, the garlic extract extract significantly inhibited the glutamate-induced HT22 cell apoptosis as compared to the ungerminated fresh garnet extract.
2-3. Assessment of inhibition of intracellular reactive oxygen species (ROS) production
In order to confirm the neuronal cell protection activity of the garlic extract, the activity of inhibiting glutamate - induced production of reactive oxygen species (ROS) was measured.
Specifically, 2 × 10 3 HT22 cells were plated on 96-well assay-bottom plates and cultured for 24 hours. After the cells were attached, each sample and 5 mM of glutamate was added and incubated for 6 hours. The cells are washed with PBS and incubated with 2 ', 7'-dichlorofluorescein diacetate (DCFDA; 2', 7'-dichlorofluorescein diacetate) dissolved in DMEM for 30 minutes and washed again with PBS. Fluorescence produced by the reaction of reactive oxygen species with dichlorofluorescein (DCF) was confirmed by fluorescence microscopy (Eclipse 80i, Nikon, Tokyo, Japan). The results are shown in Fig.
As shown in FIG. 5, the garlic extract of the present invention effectively inhibited the production of glutamate-induced reactive oxygen species as compared with the non-germinated fresh garlic extract. In particular, the garlic shoot extract on the 4th to 5th day of germination produced the best active oxygen species Inhibitory activity.
Hereinafter, a pharmaceutical composition for preventing or treating brain diseases containing the extract of garlic sprouts of the present invention as an active ingredient and a preparation example of a food composition for preventing or ameliorating brain diseases will be described, but the present invention is not limited thereto, .
Formulation Example 1. Preparation of pharmaceutical preparations
1. Manufacturing of powder
Garlic sprout extract 20 mg
The above components are mixed and filled in airtight bags to prepare powders.
2. Preparation of tablets
Garlic bud extract 10 mg
After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.
3. Preparation of capsules
Garlic bud extract 10 mg
Lactose 14.8 mg
Magnesium stearate 0.2 mg
The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.
4. Preparation of injections
Garlic bud extract 10 mg
180 mg mannitol
Sterile sterilized water for injection 2974 mg
Na 2 HPO 4 .2H 2 O 26 mg
(2 ml) per 1 ampoule in accordance with the usual injection preparation method.
5. Manufacture of liquids
Garlic sprout extract 20 mg
10 g per isomer
5 g mannitol
Purified water quantity
Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.
Formulation Example 2. Preparation of food preparation
1. Manufacture of health food
Garlic sprout extract 100 mg
Vitamin mixture quantity
70 g of vitamin A acetate
Vitamin E 1.0 mg
Vitamin B1 0.13 mg
0.15 mg of vitamin B2
Vitamin B6 0.5 mg
0.2 g of vitamin B12
Biotin 10 g
Nicotinic acid amide 1.7 mg
Folate 50 g
Calcium pantothenate 0.5 mg
Mineral mixture quantity
1.75 mg of ferrous sulfate
0.82 mg of zinc oxide
Magnesium carbonate 25.3 mg
Potassium monophosphate 15 mg
Secondary calcium phosphate 55 mg
Potassium citrate 90 mg
Magnesium chloride 24.8 mg
Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.
2. Manufacture of health drinks
Garlic sprout extract 100 mg
Vitamin C 15 g
Vitamin E (powder) 100 g
19.75 g of ferrous lactate
3.5 g of zinc oxide
Nicotinic acid amide 3.5 g
Vitamin A 0.2 g
Vitamin B1 0.25 g
Vitamin B2 0.3g
Water quantification
The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The resulting solution was filtered and sterilized in a sterilized 2 L container, ≪ / RTI >
Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.
Claims (5)
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3804709A4 (en) * | 2018-06-07 | 2022-02-23 | Wakunaga Pharmaceutical Co., Ltd. | Autophagy activating agent |
-
2014
- 2014-01-29 KR KR1020140011542A patent/KR20150090646A/en not_active Application Discontinuation
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3804709A4 (en) * | 2018-06-07 | 2022-02-23 | Wakunaga Pharmaceutical Co., Ltd. | Autophagy activating agent |
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