WO2020179982A1 - Composition containing broccoli sprouts as active ingredient for improving cognitive functions or memory - Google Patents

Composition containing broccoli sprouts as active ingredient for improving cognitive functions or memory Download PDF

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WO2020179982A1
WO2020179982A1 PCT/KR2019/013692 KR2019013692W WO2020179982A1 WO 2020179982 A1 WO2020179982 A1 WO 2020179982A1 KR 2019013692 W KR2019013692 W KR 2019013692W WO 2020179982 A1 WO2020179982 A1 WO 2020179982A1
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Prior art keywords
electric field
memory
composition
broccoli
broccoli sprouts
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PCT/KR2019/013692
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French (fr)
Korean (ko)
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최혁준
박용운
김선여
조교희
수베디라리타
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주식회사 비케이바이오
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • 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
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/322—Foods, ingredients or supplements having a functional effect on health having an effect on the health of the nervous system or on mental function
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2300/00—Processes
    • A23V2300/12—Electrical treatment, e.g. electrolysis, electrical field treatment, with or without heating effect
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material

Definitions

  • the present invention relates to a health functional food composition or a pharmaceutical composition comprising broccoli sprouts having improved cognitive function or memory improvement effect as an active ingredient.
  • the present invention relates to a method for producing broccoli sprouts with improved cognitive function or memory improvement effect.
  • AD Alzheimer's disease
  • a type of senile dementia is caused by damage to acetylcholine-related nerve cells at the base of the cerebrum, it is muscarinic.
  • Drugs that can enhance the function of acetylcholinergic neurons according to various mechanisms of action such as agonists against muscarine acetylcholine receptors, acetylcholine production promoters, and acetylcholinesterase (AChE) inhibitors.
  • AChE acetylcholinesterase
  • An object of the present invention is to provide a method for producing broccoli sprouts with improved cognitive function or memory improvement effect using high-voltage pulsed electric field treatment, and a health functional food composition or pharmaceutical composition comprising broccoli sprouts prepared therefrom.
  • an object of the present invention is to provide a use of a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) as an active ingredient for improving cognitive decline, or for manufacturing a drug for preventing or treating cognitive dysfunction.
  • PEF high voltage pulse electric field
  • an object of the present invention is to provide a method for preventing or treating cognitive dysfunction comprising administering a composition comprising broccoli sprouts treated with high voltage pulse electric field (PEF) to humans or non-human animals.
  • a composition comprising broccoli sprouts treated with high voltage pulse electric field (PEF) to humans or non-human animals.
  • the present invention is to provide a method for improving the cognitive function or memory of an individual, comprising administering a composition containing broccoli sprouts treated with a high voltage pulsed electric field (PEF) to a human or non-human animal To do.
  • a composition containing broccoli sprouts treated with a high voltage pulsed electric field (PEF) to a human or non-human animal To do.
  • the present invention provides a health functional food composition for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
  • PEF high voltage pulse electric field
  • the broccoli sprout may be at least one selected from the group consisting of raw materials, dried products, pulverized products, and extracts.
  • the high voltage pulsed electric field treatment is performed in a liquid medium containing broccoli sprouts with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ for 0.1 to 100 seconds. It can be done 50 times.
  • the high voltage pulse electric field treatment may be performed at 1.5 to 2.5 kV/cm and energy 3 to 10 kJ.
  • broccoli sprouts contained in the liquid medium may be raw materials or cuts thereof that have not been pulverized or crushed.
  • the liquid medium may be water, alcohol, or a mixture thereof.
  • the composition may inhibit the activity of acetylcholinesterase.
  • the composition may be prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, and beverage.
  • the present invention provides a pharmaceutical composition for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
  • PET high voltage pulse electric field
  • the composition may exhibit an acetylcholinesterase inhibitory effect and a long-term memory improvement effect.
  • the present invention also provides a use of a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF) as an active ingredient for improving cognitive decline or for manufacturing a drug for preventing or treating cognitive function disorders.
  • a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF) as an active ingredient for improving cognitive decline or for manufacturing a drug for preventing or treating cognitive function disorders.
  • PEF pulsed electric field
  • the cognitive decline is at least one selected from the group consisting of memory loss, cognitive decline, discrimination and forgetfulness, and the cognitive dysfunction is forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, It may be a degenerative brain disease selected from the group consisting of Peak's disease and Huntington's disease.
  • the present invention provides a method for preventing or treating cognitive dysfunction comprising administering a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) to humans or non-human animals.
  • a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF)
  • the present invention provides a method for improving cognitive function or memory of an individual, comprising administering a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF) to a human or non-human animal.
  • a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF)
  • the present invention comprises the steps of placing broccoli sprouts in a batch or continuous processing container containing a liquid medium; And performing 1 to 50 times for 0.1 to 100 seconds with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ in the liquid medium; containing, improving cognitive function or memory It provides a method for producing enhanced broccoli sprouts.
  • composition containing broccoli sprouts treated with high-voltage pulsed electric field of the present invention as an active ingredient has excellent acetylcholine-degrading enzyme inhibitory effect and long-term memory improvement effect, thereby improving cognitive function or memory, improving learning ability, and treating degenerative brain diseases including senile dementia. And is useful for prevention.
  • FIG. 2 is a graph showing the HPLC results of FIG. 1.
  • Figure 3 is a graph showing the evaluation of cognitive function or memory improvement efficacy through passive avoidance frequency according to oral administration of broccoli samples according to Examples and Comparative Examples in a scopolamine-induced mouse model.
  • the present invention provides a health functional food composition for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
  • PEF high voltage pulse electric field
  • the broccoli sprouts treated with the high-voltage pulsed electric field were sulforaphane itself, the same amount as the amount of sulforaphane contained in the sprout in a passive avoidance test using scopolamine-induced memory decay animals, and high-voltage pulses.
  • ESP inhibition and myrosinase enzyme activity treatment after electric field-treated broccoli adult or high-voltage pulse electric field treatment shows a synergistic effect than administration of broccoli sprouts with higher sulforaphane content, improving cognitive function, improving memory, improving learning ability, and degenerative It was confirmed that it is useful for the treatment and prevention of brain diseases.
  • ESP Epidermal protein catalyzes the formation of sulforaphane nitrile in broccoli, and this alternative reaction pathway competes with the reaction pathway that makes sulforaphane, so the rate of conversion from glucorapanine to sulforaphane is present in broccoli with myrosinase. Which is reduced by the ESP. Therefore, when the ESP is inactivated, it is possible to increase the content of sulforaphane in broccoli. Furthermore, if the ESP is inactivated and then the myrosinase enzyme is activated, the sulforaphane content can be further increased.
  • One of the methods of inactivating the ESP is heating to a temperature of 50°C or higher.
  • the present inventors found that broccoli sprouts treated with a high voltage pulsed electric field had a significantly higher cognitive function or memory improvement effect while having a lower sulforaphane content compared to broccoli sprouts treated with a high voltage pulsed electric field treatment, ESP inactivation and myrosinase enzyme activation treatment.
  • the present invention has been completed.
  • the broccoli sprouts treated with the high-voltage pulsed electric field are performed 1 to 50 times in a liquid medium containing broccoli sprouts with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ for 0.1 to 100 seconds. It may be manufactured by.
  • the high voltage pulse electric field treatment is preferably performed at 1.5 to 2.5 kV/cm and energy 3 to 10 kJ.
  • the broccoli sprouts contained in the liquid medium prior to the high voltage pulse electric field treatment may be raw materials or cuts thereof that have not been pulverized or ground.
  • the liquid medium is water, ethanol, or a mixture thereof.
  • Organic acids such as acetic acid, citric acid, and lactic acid, or inorganic salts such as salt may be further added to the liquid medium.
  • the broccoli sprouts after the high voltage pulse electric field treatment may be at least one selected from the group consisting of raw materials, dried products, pulverized products, and extracts. That is, the broccoli sprout itself subjected to high voltage pulse electric field treatment, or may be subjected to a high voltage pulse electric field treatment and then a drying process, a pulverization process, a crushing process after drying, a drying process after crushing, or an extraction process.
  • the extract may be extracted using a solvent selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof, preferably water, methanol, ethanol, or butanol, and more preferably Is extracted with reflux cooling using 60 to 80% (v/v) methanol as a solvent.
  • the extract is intended to contain any one of an extract obtained by an extraction treatment, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a crude product or a purified product.
  • the broccoli sprouts treated with the high voltage pulse electric field or a health functional food composition including the same as an active ingredient may inhibit the activity of acetylcholinesterase.
  • the health functional food composition may be prepared in various forms according to conventional methods known in the art, for example, prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrup, and beverages. Can be.
  • the content of broccoli sprouts treated with the high voltage pulse electric field in the health functional food composition may be contained in an amount of 0.1 to 100% by weight, preferably 0.2 to 80% by weight, more preferably, based on the total weight of the composition. May be contained in an amount of 0.3 to 60% by weight.
  • the content of the broccoli sprout is less than the above range, there is a problem that it is difficult to expect the effect of the broccoli sprout.
  • the composition of the present invention When used as a health drink, it may contain various flavoring agents or natural carbohydrates as an additional component, like a normal drink.
  • the natural carbohydrates described above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as texttrin and cyclotenstrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used.
  • the ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention.
  • the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal focus agents, pH adjusters, stabilizers, preservatives, glycerin, alcohols , Carbonated beverages used in carbonated beverages, and the like.
  • the composition of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients may be used independently or in combination. The proportion of these additives is not very important, but the composition of the present invention is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.
  • the present invention also provides a pharmaceutical composition for improving cognitive function or memory, using broccoli sprouts treated with the high voltage pulse electric field (PEF) as an active ingredient.
  • PEF high voltage pulse electric field
  • the degenerative brain diseases that cause cognitive function or memory degeneration include Alzheimer's disease, dementia, Parkinson's disease, progressive supranuclear palsy, multiple system strophy, and olive nucleus- Olivopontocerebellar atrophy (OPCA), Shy-Drager syndrome, striatonigral degeneration, Huntington's disease, amyotrophic lateral sclerosis (ALS).
  • ALS amyotrophic lateral sclerosis
  • the pharmaceutical composition of the present invention is more preferable for the prevention or treatment of forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, Peak's disease, and Huntington's disease among the degenerative brain diseases.
  • the pharmaceutical composition for improving cognitive function or memory of the present invention or the pharmaceutical composition for preventing or treating degenerative cranial nerve disorders contains 0.02 to 80% by weight of broccoli sprouts treated with the high voltage pulse electric field (PEF) based on the total weight of the pharmaceutical composition, preferably It may contain from 0.02 to 50% by weight.
  • PEF high voltage pulse electric field
  • the pharmaceutical composition of the present invention exhibits an acetylcholinesterase inhibitory effect and a long-term memory improvement effect.
  • the pharmaceutical composition of the present invention may further include suitable carriers, excipients, and diluents, which are commonly used in the preparation of pharmaceutical compositions.
  • the pharmaceutical composition according to the present invention is formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.
  • oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.
  • Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention include lactose, textrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, and methylhydroxybenzo mixtures.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient in the pharmaceutical composition, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. You can mix and prepare. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, suspensions, emulsions, freeze-dried preparations, preservatives Etc. may be included.
  • Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories.
  • Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used as the non-aqueous solvent and suspension.
  • Witapsol, Macrogol, Tween 61, cacao butter, laurazine, glycerogelatin, and the like may be used as a base for the suppository.
  • the preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the degree of disease, the form of the drug, the route and duration of administration, but may be appropriately selected by those skilled in the art.
  • broccoli sprouts treated with a high voltage pulsed electric field (PEF) of the present invention are preferably administered at 0.0001 to 100 mg/kg per day, preferably 0.001 to 100 mg/kg. Administration may be administered once a day, or may be divided several times. The above dosage does not in any way limit the scope of the present invention.
  • the pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, and humans by various routes. All modes of administration can be expected and may be administered by, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intracerebroventricular injection.
  • the present invention provides a pharmaceutical composition for animals other than humans for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
  • PET high voltage pulse electric field
  • the present invention provides a novel use of a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) as an active ingredient for improving cognitive decline or for manufacturing a drug for preventing or treating cognitive function disorders.
  • a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) as an active ingredient for improving cognitive decline or for manufacturing a drug for preventing or treating cognitive function disorders.
  • PEF high voltage pulse electric field
  • the cognitive decline may be one or more selected from the group consisting of memory loss, cognitive decline, discrimination and forgetfulness.
  • the cognitive dysfunction may be a degenerative brain disease selected from the group consisting of forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, Peak's disease, and Huntington's disease.
  • the present invention provides a method for preventing or treating cognitive dysfunction comprising administering a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) to humans or non-human animals.
  • a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF)
  • the present invention provides a method for improving cognitive function or memory of an individual, comprising administering a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF) to a human or non-human animal.
  • a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF)
  • a composition comprising broccoli sprouts treated with the high voltage pulsed electric field (PEF) may be administered by various routes.
  • the method of administration may be, for example, oral administration, or may be administered by rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
  • composition according to the present invention can be safely prescribed to exhibit an effective therapeutic effect against cognitive decline, cognitive impairment, and memory loss, and by using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient, acetylcholine decomposition It exhibits an enzyme inhibitory effect and/or an effect of improving long-term memory, thereby improving cognitive function or memory, or may be usefully used to treat or prevent cognitive dysfunction.
  • PEF high voltage pulse electric field
  • the cognitive dysfunction includes degenerative brain diseases selected from the group consisting of forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, Peak's disease, and Huntington's disease, and is not particularly limited thereto, and a more specific description of the pharmaceutical composition I will refer to.
  • the amount of use of the drug, veterinary pharmaceutical composition, or veterinary drug may vary depending on the age, sex, and weight of the patient or the animal to be treated. It will depend on the route and the nature of the therapeutic agent used.
  • the medicine, veterinary pharmaceutical composition, or veterinary medicine is appropriately selected according to the degree of absorption of the active ingredient in the body, excretion rate, age and weight, sex and condition of the patient or the animal to be treated, and the severity of the disease to be treated, but generally It is preferable to administer 0.001-100 mg/kg once.
  • the unit dosage form formulation thus formulated may be administered several times at regular time intervals as needed.
  • the medicine, veterinary pharmaceutical composition, or veterinary drug may be administered individually as a prophylactic or therapeutic agent, or may be administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent.
  • the present invention comprises the steps of placing broccoli sprouts in a batch or continuous processing container containing a liquid medium; And performing 1 to 50 times for 0.1 to 100 seconds with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ in the liquid medium; containing, improving cognitive function or memory It provides a method for producing enhanced broccoli sprouts.
  • Broccoli sprouts were purchased and used at Jeju Techno Park.
  • the high voltage pulse electric field treatment device was used as the model name ELCRACK HVP5 of German Institute of Food Technology (Germany).
  • the raw material of fresh broccoli sprouts itself was placed in a batch-type treatment container containing water of a high-voltage pulse electric field treatment device and treated with a pulse electric field once under the conditions of an electric field of 2.1 kV/cm, a frequency of 100 Hz, an energy of 7 kJ, and a treatment time of 3 seconds.
  • Example 2 It was carried out in the same manner as in Example 1, but was subjected to a single pulse electric field under the conditions of an electric field of 2.1 kV/cm, a frequency of 100 Hz, an energy of 4 kJ, and a treatment time of 3 seconds.
  • the broccoli adult body heated in the bath was cooled to 37° C. and allowed to stand for 2 hours to induce a myrosinase enzyme reaction so that the gluolapanin component was converted to sulforaphane.
  • the cut itself was put into a batch-type treatment container containing water of a high-voltage pulse electric field treatment device, and an electric field of 2.1 kV/cm, a frequency of 100 Hz, and an energy of 7 kJ and One pulse electric field treatment was performed under the treatment conditions of a treatment time of 3 seconds.
  • the broccoli adult body heated in the bath was cooled to 37° C. and allowed to stand for 2 hours to induce a myrosinase enzyme reaction so that the gluolapanin component was converted to sulforaphane.
  • Broccoli of Examples 1 and 2 and Comparative Examples 1 to 6 were prepared in a form suitable for the following experiment. First, the broccoli of Examples 1 and 2 and Comparative Examples 1 to 6 were lyophilized to powder, and then 400 ml of 70% methanol was added to 10 g of the dry powder of the samples, stirred for 3 hours, and the extracted extract was concentrated under reduced pressure. After lyophilization was performed to prepare a concentrated powder, and subsequent experiments were conducted.
  • UPLC-MS/MS manufactured by UPLC-MS/MS (manufacturer Waters) to prepare a standard quantification curve, and a sample obtained by dissolving 1 g of the concentrated powder of the samples in 1 ml of methanol was prepared by UPLC-MS.
  • the sulforaphane content was analyzed by /MS and is shown in Table 1 below.
  • UPLC system is a system consisting of Acquity UPLC system (Waters corporation, milford, MA), binary solvent manager, and sample manager, MS system is Waters Quattro Premier XE Tandem MS (Micromass UK limited), software is Masslynx V4.1. .
  • FIG. 1 is an HPLC result for better comparing the sulforaphane content of broccoli according to Examples and Comparative Examples
  • FIG. 2 is a graph showing the HPLC result.
  • the broccoli sprout has the highest content of sulforaphane when the broccoli sprouts are treated with a high-voltage pulsed electric field as well as ESP inhibition and myrosinase activation treatment.
  • the passive evasion experiment is one of the fear-motivated avoidance experiments in which the mouse learns not to enter the door of the dark room again by remembering the memory of the previous electrical stimulation in the dark room. This is an experiment that evaluates memory by measuring the time hesitated to enter the compartment previously subjected to electrical stimulation and boiling the measured time as an index of memory ability.
  • mice 6-week-old mice were supplied with ICR mice (26-28 g) from Orient Co., Ltd. (Seoul, Korea) for about 5 days to acclimate to an animal laboratory, and water and feed were freely ingested.
  • the temperature (23 ⁇ 2 °C), humidity (50 ⁇ 10%) and the contrast period (12 hours) were automatically adjusted.
  • the candidate sample was orally administered 1 hour before the start of the experiment, and 1.2 mg/kg of scopolamine was administered intraperitoneally 30 minutes before the start of the experiment.
  • the experiment was conducted at the point of time when the efficacy of the two drugs was polarized and overlapped, and statistical processing was performed by means of Mean ⁇ SEM, one way analysis of variance (ANOVA) and T-test analysis.
  • Figure 3 is a graph showing the evaluation of cognitive function or memory improvement efficacy through passive avoidance frequency according to oral administration of broccoli samples according to Examples and Comparative Examples in a scopolamine-induced mouse model.
  • mice 6-week-old mice were supplied with ICR mice (26-28 g) from Orient Co., Ltd. (Seoul, Korea) and adapted to the animal laboratory for about 5 days, and water and feed were freely ingested.
  • the temperature (23 ⁇ 2 °C), humidity (50 ⁇ 10%) and the contrast period (12 hours) were automatically adjusted.
  • donepezil As a positive control, 5 mg/kg of donepezil, which has already been used clinically as a treatment for improving memory, was administered. Scopolamine and donepezil were purchased and used from Sigma-Aldrich Chemistry Co., and other reagents were commercially available top grades.
  • the candidate sample was orally administered 1 hour before the start of the experiment, and 1.2 mg/kg of scopolamine was administered intraperitoneally 30 minutes before the start of the experiment.
  • the experiment was conducted at the point of time when the efficacy of the two drugs was polarized and overlapped, and statistical processing was performed by means of Mean ⁇ SEM, one way analysis of variance (ANOVA) and T-test analysis.
  • mice were prepared as 11 groups (4 mice per group) of the drug administration group and controls 1 and 2 to which the sulforaphane samples of Examples and Comparative Examples were administered.
  • the samples according to the sulforaphane, Examples and Comparative Examples were administered to the drug administration group at the dose determined in Table 2 (mg/Kg). Meanwhile, 0.15 mL of PBS was administered to the control group 1, and donepezil was administered at a dose of 5 mg/Kg to the control group 2.
  • the brains of the experimental animals were excised and homogenized, and the cell lysate was centrifuged at 1,000 rpm for 10 minutes, and then an enzyme activity evaluation test was performed on the supernatant.
  • a reaction solution 140 ⁇ l sodium phosphate buffer (pH 8.0), 40 ⁇ l brain test solution, 10 ⁇ l DTNB (dithiol nitrobenzoic acid)), 10 ⁇ l acetylcholineiodide at room temperature.
  • neostigmine a reaction stopping reagent
  • tablets are prepared by tableting according to a conventional tablet preparation method.
  • the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.
  • Vitamin A acetate 70 ⁇ g
  • Vitamin B12 0.2 ⁇ g
  • composition ratio of the vitamin and mineral mixture is relatively suitable for health functional foods, but it is also possible to arbitrarily modify the composition, and the above ingredients are mixed according to a general health functional food manufacturing method. Then, the granules can be prepared and used for preparing a health functional food composition according to a conventional method.
  • the resulting solution After mixing the above ingredients according to a normal health drink manufacturing method, stirring and heating the resulting solution at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and stored in a refrigerator It is used to prepare the functional beverage composition of the invention.

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Abstract

The present invention relates to a health functional food composition and a pharmaceutical composition each for improving cognitive functions or memory, the compositions each containing high-voltage pulsed electric field-treated broccoli sprouts as an active ingredient. The compositions of the present invention have excellent acetylcholine esterase inhibitory effect and long-term memory improving effect, and thus are effective in the improvement of cognitive functions or memory, the improvement of learning ability, and the treatment and prevention of degenerative brain diseases including senile dementia.

Description

브로콜리 새싹을 유효성분으로 하는 인지기능 또는 기억력 개선용 조성물Composition for improving cognitive function or memory using broccoli sprout as an active ingredient
본 발명은 인지기능 또는 기억력 개선 효과가 증진된 브로콜리 새싹을 유효성분으로 하는 건강기능식품 조성물 또는 약학 조성물에 관한 것이다. The present invention relates to a health functional food composition or a pharmaceutical composition comprising broccoli sprouts having improved cognitive function or memory improvement effect as an active ingredient.
또한 본 발명은 인지기능 또는 기억력 개선 효과가 증진된 브로콜리 새싹의 제조방법에 관한 것이다.In addition, the present invention relates to a method for producing broccoli sprouts with improved cognitive function or memory improvement effect.
현대인에게 기억력은 급변하는 생활에서 그 중요성이 증대되고 있으며, 사회적으로 학습량이 많은 청소년에서부터 노년에 이르기까지 주요 관심 대상이 되고 있다.For modern people, memory is increasing in importance in a rapidly changing life, and it has become a major object of interest from adolescents with a large amount of learning socially to old age.
또한, 최근 생활수준의 향상과 의학기술의 발달에 따라 노령 인구가 증가하여 인구의 고령화 현상이 두드러지고 있으며, 조직사회의 치열한 경쟁화에 따른 극심한 스트레스로 인한 성인병의 발병률 또한 급격한 증가 추세를 보이고 있다. 사망원인의 경우 과거 주원인이었던 감염형 질환에서 대표적인 성인병인 순환기계 질환, 암 등에 의한 사망이 전체의 절반을 넘기기에 이르렀다. 이에 건강한 삶을 누리고자 하는 '웰빙(wellbeing)' 이 하나의 큰 관심사로 대두하게 되고 장수에 대한 욕구 증가로 일상생활을 통한 성인병 및 노인성 질환에 대한 예방에 관심이 고조되고 있다. 최근 식품성분에 대해 새로운 관점에서의 체계적인 연구가 발전함에 따라 식품기원의 특정 성분들이 인체의 신경계, 생체 방어계, 세포분화(cell differentiation) 등 각종 생리기능 조절계 등에 작용하여 직·간접적으로 생체조절기능 효과를 나타낸다는 사실들이 밝혀지고 있다. 이들 특정성분들에 의한 생체 조절계 및 생체 방어계에 대한 적절한 조절이 가능하게 된다면 그것은 가장 자연스러운 건강유지를 위한 최적의 수단이 될 수 있음을 의미한다. 식품에 도입되고 있는 생체 조절성 3차 기능은 '질병 발병의 전 단계에서 식품을 통하여 예방한다'는 적극적인 식품의 개념으로, 향후 고령화 사회에서 건강기능식품 및 식품의약품의 수요가 합성의약품을 상회할 것으로 예측되고 있기도 하다. 또한 2011년도 교육과학기술부의 '기초연구 100대 미래유망 분야' 에 의하면 '뇌신경기능과 질환', '신경계 질환' 및 '의약품 소재 추출 및 합성' 분야 등이 선정되었다.In addition, the aging of the population is remarkable due to the increase in the aging population due to the recent improvement in living standards and the development of medical technology, and the incidence of adult diseases due to extreme stress due to fierce competition in the organizational society is also showing a sharp increase. . As for the cause of death, more than half of the deaths from infectious diseases, which were the main causes in the past, from circulatory system diseases and cancer, which are representative adult diseases, have reached. Accordingly,'wellbeing' to enjoy a healthy life has emerged as a major concern, and as the desire for longevity increases, interest in preventing adult diseases and senile diseases through daily life is increasing. With the recent development of systematic research on food ingredients from a new perspective, certain ingredients of food origin act on various physiological function control systems such as the nervous system, biological defense system, and cell differentiation of the human body, thereby controlling the body directly or indirectly. It has been found to be effective. If appropriate control of the biological control system and the biological defense system by these specific components is possible, it means that it can be an optimal means for maintaining the most natural health. The bioregulatory tertiary function introduced in food is the concept of active food that'prevents through food in the pre-disease stage', and the demand for health functional foods and food drugs in the aging society will exceed synthetic drugs. It is also expected to be. In addition, according to the 2011 Ministry of Education, Science and Technology's '100 basic research areas','cranial nerve function and disease','nervous system disease', and'extraction and synthesis of pharmaceutical materials' were selected.
노인성 치매의 일종인 알츠하이머병(Alzheimer's disease, AD)으로부터 유발되는 기억력 및 학습능력의 상실 등 인지 기능 장애는 대뇌 기저부의 아세틸콜린(Acethylcholine)성 신경세포의 손상에서 비롯된다는 연구결과에 의해 무스카린성 아세틸콜린 수용체(muscarine acetylcholine receptor)에 대한 효능제(agonist), 아세틸콜린 생성촉진제, 아세틸콜린 분해효소(Acetylcholinesterase, AChE) 저해제 등 여러 가지 작용기전에 따라 아세틸콜린성 신경세포의 기능을 강화시켜줄 수 있는 약물들이 개발되어 왔다. 실제로 현재 각국에서 사용되고 있는 알츠하이머병 치료제는 이러한 아세틸콜린 분해효소 억제제가 대부분이며, 타크린(tacrine)과 도네페질(donepezil) 등이 이에 속한다. 이 밖에도 리바스티그민(rivastigmin), 갈라타민(galatamine) 등이 사용되고 있다.According to a study that shows that cognitive dysfunction, such as loss of memory and learning ability, caused by Alzheimer's disease (AD), a type of senile dementia, is caused by damage to acetylcholine-related nerve cells at the base of the cerebrum, it is muscarinic. Drugs that can enhance the function of acetylcholinergic neurons according to various mechanisms of action, such as agonists against muscarine acetylcholine receptors, acetylcholine production promoters, and acetylcholinesterase (AChE) inhibitors. Has been developed. In fact, most of Alzheimer's disease treatments currently used in each country are acetylcholine-degrading enzyme inhibitors, and tacrine and donepezil belong to this. In addition, rivastigmin and galatamine are used.
오늘날은, 퇴행성 뇌신경 질환으로 인한 인지기능 또는 기억능력 저하 현상 개선을 위한 더욱 효과적인 제제(건강기능성 식품 소재 포함)의 개발 수요가 어느 때보다 높은 시기이다. 결국 퇴행성 뇌신경 질환, 특히 치매 치료에 탁월한 효과를 나타내고 부작용이 없는 치료제에 대한 연구개발이 절실한 실정이다.Today, the demand for the development of more effective agents (including health-functional food ingredients) for improving cognitive function or memory decline caused by degenerative neurological diseases is higher than ever. In the end, there is an urgent need for research and development on therapeutic agents that exhibit excellent effects in the treatment of degenerative neurological diseases, especially dementia, and have no side effects.
본 발명자들은 인지기능 또는 기억력 개선에 효과가 좋은 소재에 대하여 연구하던 중에 브로콜리 새싹에 고전압 펄스 전기장 처리를 하는 경우, 상기 새싹에 포함되어 있는 설포라판 함량과 동일량의 설포라판 자체, 고전압 펄스 전기장 처리된 브로콜리 성체 또는 고전압 펄스 전기장 처리 후 ESP 저해 및 미로시나제 효소 활성 처리하여 설포라판 함량이 더욱 높은 브로콜리 새싹보다 인지기능 개선 또는 기억력 개선 효과가 더욱 높게 나타나 인지기능 개선, 기억력 증진, 학습능력 향상, 퇴행성 뇌질환의 치료 및 예방에 유용함을 확인하고 본 발명을 완성하게 되었다.When the present inventors are researching a material that is effective in improving cognitive function or memory, when a high-voltage pulsed electric field is applied to a broccoli sprout, the sulforaphane itself equal to the amount of sulforaphane contained in the sprout, and broccoli treated with a high-voltage pulsed electric field After treatment with adult or high voltage pulse electric field, ESP inhibition and myrosinase enzyme activity treatment have higher cognitive function improvement or memory improvement effect than broccoli sprouts with higher sulforaphane content, improving cognitive function, enhancing memory, improving learning ability, degenerative brain disease It was confirmed that it is useful in the treatment and prevention of, and the present invention was completed.
본 발명은 고전압 펄스 전기장 처리를 이용하여 인지기능 또는 기억력 개선 효과가 증진된 브로콜리 새싹의 제조방법, 그로부터 제조된 브로콜리 새싹을 포함하는 건강기능식품 조성물 또는 약학 조성물을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a method for producing broccoli sprouts with improved cognitive function or memory improvement effect using high-voltage pulsed electric field treatment, and a health functional food composition or pharmaceutical composition comprising broccoli sprouts prepared therefrom.
또한, 본 발명은 인지기능 저하 개선용, 또는 인지기능 장애 예방 또는 치료용 의약 제조를 위한, 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 포함하는 조성물의 용도를 제공하는 것을 목적으로 한다.In addition, an object of the present invention is to provide a use of a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) as an active ingredient for improving cognitive decline, or for manufacturing a drug for preventing or treating cognitive dysfunction. .
또한, 본 발명은 인간, 또는 인간을 제외한 동물에게 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물을 투여하는 단계를 포함하는 인지기능 장애의 예방 또는 치료 방법을 제공하는 것을 목적으로 한다.In addition, an object of the present invention is to provide a method for preventing or treating cognitive dysfunction comprising administering a composition comprising broccoli sprouts treated with high voltage pulse electric field (PEF) to humans or non-human animals.
또한, 본 발명은 인간, 또는 인간을 제외한 동물에게 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물을 투여하는 단계를 포함하는, 개체의 인지기능 또는 기억력을 개선시키는 방법을 제공하는 것을 목적으로 한다.In addition, the present invention is to provide a method for improving the cognitive function or memory of an individual, comprising administering a composition containing broccoli sprouts treated with a high voltage pulsed electric field (PEF) to a human or non-human animal To do.
상기한 목적을 달성하기 위하여 본 발명은 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 하는 인지기능 또는 기억력 개선용 건강기능식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a health functional food composition for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
본 발명의 일 실시예에 의하면, 상기 브로콜리 새싹은 원물, 건조물, 분쇄물, 및 추출물로 이루어진 군에서 선택된 1종 이상일 수 있다.According to an embodiment of the present invention, the broccoli sprout may be at least one selected from the group consisting of raw materials, dried products, pulverized products, and extracts.
본 발명의 일 실시예에 의하면, 상기 고전압 펄스 전기장 처리는 브로콜리 새싹이 담긴 액상 매질에 전기장 0.1 내지 10 kV/cm, 주파수 1 내지 100 Hz 및 에너지 1 내지 20 kJ로 0.1초 내지 100 초 동안 1 내지 50회 수행하는 것일 수 있다.According to an embodiment of the present invention, the high voltage pulsed electric field treatment is performed in a liquid medium containing broccoli sprouts with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ for 0.1 to 100 seconds. It can be done 50 times.
본 발명의 일 실시예에 의하면, 상기 고전압 펄스 전기장 처리는 1.5 내지 2.5 kV/cm 및 에너지 3 내지 10 kJ로 수행될 수 있다.According to an embodiment of the present invention, the high voltage pulse electric field treatment may be performed at 1.5 to 2.5 kV/cm and energy 3 to 10 kJ.
본 발명의 일 실시예에 의하면, 상기 액상 매질에 담긴 브로콜리 새싹은 분쇄 또는 마쇄과정을 거치지 않은 원물 또는 그 절단물일 수 있다.According to an embodiment of the present invention, broccoli sprouts contained in the liquid medium may be raw materials or cuts thereof that have not been pulverized or crushed.
본 발명의 일 실시예에 의하면, 상기 액상 매질은 물, 주정 또는 이들의 혼합액일 수 있다.According to an embodiment of the present invention, the liquid medium may be water, alcohol, or a mixture thereof.
본 발명의 일 실시예에 의하면, 상기 조성물은 아세틸콜린 분해효소(acetylcholinesterase)의 활성을 저해할 수 있다.According to an embodiment of the present invention, the composition may inhibit the activity of acetylcholinesterase.
본 발명의 일 실시예에 의하면, 상기 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 어느 하나의 제형으로 제조될 수 있다.According to an embodiment of the present invention, the composition may be prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, and beverage.
또한, 본 발명은 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 하는 인지기능 또는 기억력 개선용 약학 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
본 발명의 일 실시예에 의하면, 상기 조성물은 아세틸콜린 분해효소(acetylcholinesterase) 저해 효능 및 장기적 기억력 개선 효능을 나타낼 수 있다.According to an embodiment of the present invention, the composition may exhibit an acetylcholinesterase inhibitory effect and a long-term memory improvement effect.
본 발명은 또한, 인지기능 저하 개선용, 또는 인지기능 장애 예방 또는 치료용 의약 제조를 위한, 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 포함하는 조성물의 용도를 제공한다.The present invention also provides a use of a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF) as an active ingredient for improving cognitive decline or for manufacturing a drug for preventing or treating cognitive function disorders.
본 발명의 일 실시예에 의하면, 상기 인지기능 저하는 기억력 감퇴, 인지 감퇴, 식별력 감퇴 및 건망증으로 구성되는 군으로부터 선택되는 하나 이상이고, 상기 인지기능 장애는 건망증, 알츠하이머병, 치매, 파킨슨 병, 피크병, 및 헌팅톤병으로 이루어지는 군으로부터 선택되는 퇴행성 뇌질환일 수 있다.According to an embodiment of the present invention, the cognitive decline is at least one selected from the group consisting of memory loss, cognitive decline, discrimination and forgetfulness, and the cognitive dysfunction is forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, It may be a degenerative brain disease selected from the group consisting of Peak's disease and Huntington's disease.
또한, 본 발명은 인간, 또는 인간을 제외한 동물에게 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물을 투여하는 단계를 포함하는 인지기능 장애의 예방 또는 치료 방법을 제공한다.In addition, the present invention provides a method for preventing or treating cognitive dysfunction comprising administering a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) to humans or non-human animals.
또한, 본 발명은 인간, 또는 인간을 제외한 동물에게 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물을 투여하는 단계를 포함하는, 개체의 인지기능 또는 기억력을 개선시키는 방법을 제공한다.In addition, the present invention provides a method for improving cognitive function or memory of an individual, comprising administering a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF) to a human or non-human animal.
또한, 본 발명은 브로콜리 새싹을 액상 매질이 담긴 회분식 또는 연속식 처리 용기에 넣는 단계; 및 상기 액상 매질에 전기장 0.1 내지 10 kV/cm, 주파수 1 내지 100 Hz 및 에너지 1 내지 20 kJ로 0.1초 내지 100 초 동안 1 내지 50회 수행하는 단계;를 포함하는, 인지기능 또는 기억력 개선 효과가 증진된 브로콜리 새싹의 제조방법을 제공한다.In addition, the present invention comprises the steps of placing broccoli sprouts in a batch or continuous processing container containing a liquid medium; And performing 1 to 50 times for 0.1 to 100 seconds with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ in the liquid medium; containing, improving cognitive function or memory It provides a method for producing enhanced broccoli sprouts.
본 발명의 고전압 펄스 전기장 처리된 브로콜리 새싹을 유효성분으로 하는 조성물은 아세틸콜린 분해효소 저해 효능 및 장기적 기억력 개선 효능 이 우수하여 인지기능 또는 기억력 개선, 학습능력 향상, 노인성 치매를 비롯한 퇴행성 뇌질환의 치료 및 예방에 유용하다.The composition containing broccoli sprouts treated with high-voltage pulsed electric field of the present invention as an active ingredient has excellent acetylcholine-degrading enzyme inhibitory effect and long-term memory improvement effect, thereby improving cognitive function or memory, improving learning ability, and treating degenerative brain diseases including senile dementia. And is useful for prevention.
도 1은 실시예 및 비교예에 따른 브로콜리의 설포라판 함량을 비교하여 나타내기 위한 HPLC 결과이다.1 is an HPLC result for comparing and showing the sulforaphane content of broccoli according to Examples and Comparative Examples.
도 2는 도 1의 HPLC 결과를 그래프로 나타낸 것이다.2 is a graph showing the HPLC results of FIG. 1.
도 3은 스코폴라민 유도 마우스 모델에서 실시예 및 비교예에 따른 브로콜리 시료의 경구 투여에 따른 수동회피 빈도를 통한 인지기능 또는 기억력 개선 효능을 평가하여 나타내는 그래프이다.Figure 3 is a graph showing the evaluation of cognitive function or memory improvement efficacy through passive avoidance frequency according to oral administration of broccoli samples according to Examples and Comparative Examples in a scopolamine-induced mouse model.
이하, 본 발명을 상세하게 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 일 구현예에 따르면, 본 발명은 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 하는 인지기능 또는 기억력 개선용 건강기능식품 조성물을 제공한다. According to one embodiment of the present invention, the present invention provides a health functional food composition for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
상기의 고전압 펄스 전기장 처리된 브로콜리 새싹은 스코폴라민(scopolamine)으로 유도된 기억력 감퇴동물을 이용한 수동회피시험(passive avoidance test)에서 상기 새싹에 포함되어 있는 설포라판 함량과 동일량의 설포라판 자체, 고전압 펄스 전기장 처리된 브로콜리 성체 또는 고전압 펄스 전기장 처리 후 ESP 저해 및 미로시나제 효소 활성 처리하여 설포라판 함량이 더욱 높은 브로콜리 새싹을 투여하는 것보다도 상승적인 효과가 나타나 인지기능 개선, 기억력 증진, 학습능력 향상, 퇴행성 뇌질환의 치료 및 예방에 유용함을 확인하였다.The broccoli sprouts treated with the high-voltage pulsed electric field were sulforaphane itself, the same amount as the amount of sulforaphane contained in the sprout in a passive avoidance test using scopolamine-induced memory decay animals, and high-voltage pulses. ESP inhibition and myrosinase enzyme activity treatment after electric field-treated broccoli adult or high-voltage pulse electric field treatment shows a synergistic effect than administration of broccoli sprouts with higher sulforaphane content, improving cognitive function, improving memory, improving learning ability, and degenerative It was confirmed that it is useful for the treatment and prevention of brain diseases.
한편, ESP(Epithiospecifer protein)는 브로콜리 내에서 설포라판 니트릴의 형성을 촉매하고 이 대체 반응 경로는 설포라판을 만드는 반응 경로와 경합하므로, 글루코라파닌으로부터 설포라판으로 전환되는 비율은 브로콜리 내에 미로시나제와 함께 존재하는 상기 ESP에 의해 감소된다. 따라서, 상기 ESP를 불활성화시키는 경우 브로콜리 내 설포라판 함량을 증가시킬 수 있게 된다. 더 나아가, 상기 ESP를 불활성화시킨 후 미로시나제 효소를 활성화시키면 설포라판 함량을 더욱 증가시킬 수 있게 된다. 상기 ESP를 불활성화시키는 방법 중 하나는 50 ℃ 이상의 온도로 가열하는 것이다.On the other hand, ESP (Epithiospecifer protein) catalyzes the formation of sulforaphane nitrile in broccoli, and this alternative reaction pathway competes with the reaction pathway that makes sulforaphane, so the rate of conversion from glucorapanine to sulforaphane is present in broccoli with myrosinase. Which is reduced by the ESP. Therefore, when the ESP is inactivated, it is possible to increase the content of sulforaphane in broccoli. Furthermore, if the ESP is inactivated and then the myrosinase enzyme is activated, the sulforaphane content can be further increased. One of the methods of inactivating the ESP is heating to a temperature of 50°C or higher.
본 발명자들은 고전압 펄스 전기장 처리된 브로콜리 새싹이 고전압 펄스 전기장 처리, ESP 불활성화 및 미로시나제 효소 활성화 처리를 한 브로콜리 새싹에 비해 설포라판 함량은 더욱 낮으면서도 인지기능 또는 기억력 개선 효과가 현저히 높은 것을 발견하고 본 발명을 완성하였다.The present inventors found that broccoli sprouts treated with a high voltage pulsed electric field had a significantly higher cognitive function or memory improvement effect while having a lower sulforaphane content compared to broccoli sprouts treated with a high voltage pulsed electric field treatment, ESP inactivation and myrosinase enzyme activation treatment. The present invention has been completed.
본 발명에서 상기 고전압 펄스 전기장 처리된 브로콜리 새싹은 브로콜리 새싹이 담긴 액상 매질에 전기장 0.1 내지 10 kV/cm, 주파수 1 내지 100 Hz 및 에너지 1 내지 20 kJ로 0.1초 내지 100 초 동안 1 내지 50회 수행하여 제조된 것일 수 있다.In the present invention, the broccoli sprouts treated with the high-voltage pulsed electric field are performed 1 to 50 times in a liquid medium containing broccoli sprouts with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ for 0.1 to 100 seconds. It may be manufactured by.
상기 고전압 펄스 전기장 처리는 1.5 내지 2.5 kV/cm 및 에너지 3 내지 10 kJ로 수행하는 것이 바람직하다.The high voltage pulse electric field treatment is preferably performed at 1.5 to 2.5 kV/cm and energy 3 to 10 kJ.
상기 고전압 펄스 전기장 처리 전의 상기 액상 매질에 담긴 브로콜리 새싹은 분쇄 또는 마쇄과정을 거치지 않은 원물이거나 그 절단물일 수 있다.The broccoli sprouts contained in the liquid medium prior to the high voltage pulse electric field treatment may be raw materials or cuts thereof that have not been pulverized or ground.
상기 액상 매질은 물, 에탄올 또는 이들의 혼합액인 것이 바람직하다.It is preferable that the liquid medium is water, ethanol, or a mixture thereof.
상기 액상 매질에는 아세트산, 구연산, 젖산 등의 유기산이나 소금 등의 무기염류가 더 첨가될 수 있다.Organic acids such as acetic acid, citric acid, and lactic acid, or inorganic salts such as salt may be further added to the liquid medium.
그리고, 상기 고전압 펄스 전기장 처리를 한 후의 브로콜리 새싹은 원물, 건조물, 분쇄물, 및 추출물로 이루어진 군에서 선택된 1종 이상일 수 있다. 즉, 고전압 펄스 전기장 처리를 한 브로콜리 새싹 원물 그 자체이거나, 고전압 펄스 전기장 처리를 한 후 건조 공정, 분쇄 공정, 건조 후 분쇄 공정, 분쇄 후 건조 공정 또는 추출 공정을 거친 것일 수 있다. 상기 추출물은 물, 탄소수 1 내지 4의 저급 알코올 및 이들의 혼합용매로 구성되는 군으로부터 선택되는 용매, 바람직하게는 물, 메탄올, 에탄올, 또는 부탄올의 용매를 이용하여 추출한 것일 수 있으며, 더 바람직하게는 60 내지 80 %(v/v) 메탄올을 용매로 이용하여 환류 냉각 추출한 것이다. 또한, 상기 추출물은 추출처리에 의해 얻어지는 추출액, 추출액의 희석액 또는 농축액, 추출액을 건조하여 얻어지는 건조물, 또는 조정제물 또는 정제물 중 어느 하나를 포함하는 것으로 한다.In addition, the broccoli sprouts after the high voltage pulse electric field treatment may be at least one selected from the group consisting of raw materials, dried products, pulverized products, and extracts. That is, the broccoli sprout itself subjected to high voltage pulse electric field treatment, or may be subjected to a high voltage pulse electric field treatment and then a drying process, a pulverization process, a crushing process after drying, a drying process after crushing, or an extraction process. The extract may be extracted using a solvent selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof, preferably water, methanol, ethanol, or butanol, and more preferably Is extracted with reflux cooling using 60 to 80% (v/v) methanol as a solvent. In addition, the extract is intended to contain any one of an extract obtained by an extraction treatment, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a crude product or a purified product.
상기 고전압 펄스 전기장 처리된 브로콜리 새싹 또는 이를 유효성분으로 포함하는 건강기능식품 조성물은 아세틸콜린 분해효소(acetylcholinesterase)의 활성을 저해할 수 있다.The broccoli sprouts treated with the high voltage pulse electric field or a health functional food composition including the same as an active ingredient may inhibit the activity of acetylcholinesterase.
상기 건강기능식품 조성물은 당 업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있고, 예를 들면 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 어느 하나의 제형으로 제조될 수 있다. 본 발명에 의하면 상기 건강기능식품 조성물에서 상기 고전압 펄스 전기장 처리된 브로콜리 새싹의 함량은 전체 조성물 총 중량에 대하여 0.1 내지 100 중량%로 함유될 수 있으며, 바람직하게는 0.2 내지 80 중량%, 더 바람직하게는 0.3 내지 60 중량%로 함유될 수 있다. 상기 브로콜리 새싹의 함량이 상기 범위 미만인 경우에는 상기 브로콜리 새싹에 의한 효과를 기대하기 어려운 문제가 있다. The health functional food composition may be prepared in various forms according to conventional methods known in the art, for example, prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrup, and beverages. Can be. According to the present invention, the content of broccoli sprouts treated with the high voltage pulse electric field in the health functional food composition may be contained in an amount of 0.1 to 100% by weight, preferably 0.2 to 80% by weight, more preferably, based on the total weight of the composition. May be contained in an amount of 0.3 to 60% by weight. When the content of the broccoli sprout is less than the above range, there is a problem that it is difficult to expect the effect of the broccoli sprout.
본 발명의 조성물을 건강음료로 사용할 경우, 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 텍스트린, 사이클로텐스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100g당 일반적으로 약 0.01~0.04g, 바람직하게는 약 0.02~0.03g이다.When the composition of the present invention is used as a health drink, it may contain various flavoring agents or natural carbohydrates as an additional component, like a normal drink. The natural carbohydrates described above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as texttrin and cyclotenstrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 중점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물은 100 중량부당 0.01~0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal focus agents, pH adjusters, stabilizers, preservatives, glycerin, alcohols , Carbonated beverages used in carbonated beverages, and the like. In addition, the composition of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients may be used independently or in combination. The proportion of these additives is not very important, but the composition of the present invention is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.
본 발명은 또한, 상기 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 하는 인지기능 또는 기억력 개선용 약학 조성물을 제공한다. The present invention also provides a pharmaceutical composition for improving cognitive function or memory, using broccoli sprouts treated with the high voltage pulse electric field (PEF) as an active ingredient.
또한, 상기 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 하는 퇴행성 뇌질환의 예방 또는 치료용 약학 조성물을 제공한다. 상기 인지기능 또는 기억력 퇴화를 유발하는 퇴행성 뇌질환에는 알츠하이머병(Alzheimer's disease), 치매, 파킨슨병(Parkinson's disease), 진행성 핵상마비(Progressive supranuclear palsy), 다계통 위축증(Multiple system strophy), 감람핵-뇌교-소뇌 위축증(Olivopontocerebellar atrophy: OPCA), 샤이-드래거 증후군(Shy-Drager syndrome), 선조체-흑질 퇴행증(Striatonigral degeneration), 헌팅톤병(Huntington's disease), 근위축성 측색 경화증(Amyotrophic lateral sclerosis: ALS), 본태성 진전증(Essential tremor), 피질-기저핵 퇴행증(Cortico-basal ganlionic degeneration), 미만성 루이 소체 질환(Diffuse Lewy body disease), 파킨슨-ALS-치매 복합증(Parkinson-ALSdementia complex of Guam) 또는 픽병(Pick's disease)이 있으나, 이에 제한되지 않는다. 본 발명의 약학 조성물은 상기 퇴행성 뇌질환 중에서 건망증, 알츠하이머병, 치매, 파킨슨 병, 피크병, 및 헌팅톤병의 예방 또는 치료에 더욱 바람직하다.In addition, it provides a pharmaceutical composition for the prevention or treatment of degenerative brain diseases using the broccoli sprouts treated with the high voltage pulse electric field (PEF) as an active ingredient. The degenerative brain diseases that cause cognitive function or memory degeneration include Alzheimer's disease, dementia, Parkinson's disease, progressive supranuclear palsy, multiple system strophy, and olive nucleus- Olivopontocerebellar atrophy (OPCA), Shy-Drager syndrome, striatonigral degeneration, Huntington's disease, amyotrophic lateral sclerosis (ALS). ), Essential tremor, Cortico-basal ganlionic degeneration, Diffuse Lewy body disease, Parkinson-ALS-dementia complex of Guam, or Pick's disease, but is not limited thereto. The pharmaceutical composition of the present invention is more preferable for the prevention or treatment of forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, Peak's disease, and Huntington's disease among the degenerative brain diseases.
본 발명의 인지기능 또는 기억력 개선용 약학 조성물 또는 퇴행성 뇌신경 질환의 예방 또는 치료용 약학 조성물은 약학조성물 총 중량에 대하여 상기 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 0.02 내지 80 중량%, 바람직하게는 0.02 내지 50 중량%로 포함할 수 있다.The pharmaceutical composition for improving cognitive function or memory of the present invention or the pharmaceutical composition for preventing or treating degenerative cranial nerve disorders contains 0.02 to 80% by weight of broccoli sprouts treated with the high voltage pulse electric field (PEF) based on the total weight of the pharmaceutical composition, preferably It may contain from 0.02 to 50% by weight.
본 발명의 약학 조성물은 아세틸콜린 분해효소(acetylcholinesterase) 저해 효능 및 장기적 기억력 개선 효능을 나타낸다.The pharmaceutical composition of the present invention exhibits an acetylcholinesterase inhibitory effect and a long-term memory improvement effect.
본 발명의 약학 조성물은 약학 조성물의 제조에 통상적으로 사용하여 적절한 담체, 부형제 및 희석제를 더 포함할 수도 있다.The pharmaceutical composition of the present invention may further include suitable carriers, excipients, and diluents, which are commonly used in the preparation of pharmaceutical compositions.
본 발명에 따른 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 본 발명의 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 텍스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조 혼합물을 들 수 있다.The pharmaceutical composition according to the present invention is formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. I can. Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention include lactose, textrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, and methylhydroxybenzo mixtures.
경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 약학 조성물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트, 수크로스 또는 락토오스, 젤라틴 등을 섞어 조제할 수 있다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 현탁제, 유제, 동결건조 제제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위탭솔, 마크로골, 트윈 61, 카카오지, 라우진지, 글리세로제라틴 등이 사용될 수 있다.Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient in the pharmaceutical composition, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. You can mix and prepare. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, suspensions, emulsions, freeze-dried preparations, preservatives Etc. may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used as the non-aqueous solvent and suspension. As a base for the suppository, Witapsol, Macrogol, Tween 61, cacao butter, laurazine, glycerogelatin, and the like may be used.
본 발명의 약학 조성물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수있다. 그러나, 바람직한 효과를 위해서, 본 발명의 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹은 1일 0.0001 내지 100㎎/kg으로, 바람직하게는 0.001 내지 100㎎/kg으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the degree of disease, the form of the drug, the route and duration of administration, but may be appropriately selected by those skilled in the art. However, for desirable effects, broccoli sprouts treated with a high voltage pulsed electric field (PEF) of the present invention are preferably administered at 0.0001 to 100 mg/kg per day, preferably 0.001 to 100 mg/kg. Administration may be administered once a day, or may be divided several times. The above dosage does not in any way limit the scope of the present invention.
본 발명의 약학 조성물은 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내(intracerebroventricular) 주사에 의해 투여될 수 있다.The pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, and humans by various routes. All modes of administration can be expected and may be administered by, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intracerebroventricular injection.
또한, 본 발명은 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 하는 인지기능 또는 기억력 개선을 위한 인간을 제외한 동물용 약학 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for animals other than humans for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
또한, 본 발명은 인지기능 저하 개선용, 또는 인지기능 장애 예방 또는 치료용 의약 제조를 위한, 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 포함하는 조성물의 신규 용도를 제공한다.In addition, the present invention provides a novel use of a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) as an active ingredient for improving cognitive decline or for manufacturing a drug for preventing or treating cognitive function disorders.
상기 인지기능 저하는 기억력 감퇴, 인지 감퇴, 식별력 감퇴 및 건망증으로 구성되는 군으로부터 선택되는 하나 이상일 수 있다. 또한, 상기 인지기능 장애는 건망증, 알츠하이머병, 치매, 파킨슨 병, 피크병, 및 헌팅톤병으로 이루어지는 군으로부터 선택되는 퇴행성 뇌질환일 수 있다.The cognitive decline may be one or more selected from the group consisting of memory loss, cognitive decline, discrimination and forgetfulness. In addition, the cognitive dysfunction may be a degenerative brain disease selected from the group consisting of forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, Peak's disease, and Huntington's disease.
또한, 본 발명은 인간, 또는 인간을 제외한 동물에게 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물을 투여하는 단계를 포함하는 인지기능 장애의 예방 또는 치료 방법을 제공한다.In addition, the present invention provides a method for preventing or treating cognitive dysfunction comprising administering a composition comprising broccoli sprouts treated with a high voltage pulse electric field (PEF) to humans or non-human animals.
또한, 본 발명은 인간, 또는 인간을 제외한 동물에게 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물을 투여하는 단계를 포함하는, 개체의 인지기능 또는 기억력을 개선시키는 방법을 제공한다.In addition, the present invention provides a method for improving cognitive function or memory of an individual, comprising administering a composition comprising broccoli sprouts treated with a high voltage pulsed electric field (PEF) to a human or non-human animal.
상기 인지기능 장애의 예방 또는 치료 방법, 또는 인지기능 또는 기억력을 개선시키는 방법에서, 상기 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물은 다양한 경로로 투여될 수 있다. 투여 방법은, 예를 들면 경구 투여이거나, 직장 또는 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내(intracerebroventricular) 주사에 의해 투여될 수 있다.In the method of preventing or treating cognitive disorders, or improving cognitive function or memory, a composition comprising broccoli sprouts treated with the high voltage pulsed electric field (PEF) may be administered by various routes. The method of administration may be, for example, oral administration, or may be administered by rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
본 발명에 따른 조성물은 인지기능 저하, 인지기능 장애 및 기억력 감퇴 등에 대해 효과적인 치료효과를 나타내도록 안전하게 처방할 수 있는 것으로, 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 함으로써, 아세틸콜린 분해효소 저해 효능 및/또는 장기적 기억력 개선 효능을 나타내어 인지기능 또는 기억력을 개선하거나, 인지기능 장애를 치료 또는 예방하는데 유용하게 사용될 수 있다.The composition according to the present invention can be safely prescribed to exhibit an effective therapeutic effect against cognitive decline, cognitive impairment, and memory loss, and by using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient, acetylcholine decomposition It exhibits an enzyme inhibitory effect and/or an effect of improving long-term memory, thereby improving cognitive function or memory, or may be usefully used to treat or prevent cognitive dysfunction.
상기 인지기능 장애는 건망증, 알츠하이머병, 치매, 파킨슨 병, 피크병, 및 헌팅톤병으로 이루어지는 군에서 선택되는 퇴행성 뇌질환을 포함하는 것으로, 특별히 이에 한정되지 않으며, 보다 구체적인 사항은 약학 조성물에 대한 설명을 참고하기로 한다.The cognitive dysfunction includes degenerative brain diseases selected from the group consisting of forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, Peak's disease, and Huntington's disease, and is not particularly limited thereto, and a more specific description of the pharmaceutical composition I will refer to.
상기 의약, 동물용 약학 조성물 또는 동물용 의약의 사용량은 환자 또는 치료대상 동물의 나이, 성별, 체중에 따라 달라질 수 있으며, 무엇보다도, 치료대상 개체의 상태, 치료 대상 질환의 특정한 카테고리 또는 종류, 투여 경로, 사용되는 치료제의 속성에 의존적일 것이다.The amount of use of the drug, veterinary pharmaceutical composition, or veterinary drug may vary depending on the age, sex, and weight of the patient or the animal to be treated. It will depend on the route and the nature of the therapeutic agent used.
상기 의약, 동물용 약학 조성물 또는 동물용 의약은 체내에서 활성성분의 흡수도, 배설속도, 환자 또는 치료대상 동물의 연령 및 체중, 성별 및 상태, 치료할 질병의 중증정도 등에 따라 적절히 선택되나, 일반적으로 1회 0.001-100 ㎎/㎏으로 투여하는 것이 바람직하다. 이렇게 제형화된 단위 투여형 제제는 필요에 따라 일정시간 간격으로 수회 투여할 수 있다.The medicine, veterinary pharmaceutical composition, or veterinary medicine is appropriately selected according to the degree of absorption of the active ingredient in the body, excretion rate, age and weight, sex and condition of the patient or the animal to be treated, and the severity of the disease to be treated, but generally It is preferable to administer 0.001-100 mg/kg once. The unit dosage form formulation thus formulated may be administered several times at regular time intervals as needed.
상기 의약, 동물용 약학 조성물 또는 동물용 의약은 개별적으로 예방제 또는 치료제로서 투여하거나 다른 치료제와 병용하여 투여될 수 있고, 종래의 치료제와는 순차적 또는 동시에 투여될 수 있다.The medicine, veterinary pharmaceutical composition, or veterinary drug may be administered individually as a prophylactic or therapeutic agent, or may be administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent.
또한, 본 발명은 브로콜리 새싹을 액상 매질이 담긴 회분식 또는 연속식 처리 용기에 넣는 단계; 및 상기 액상 매질에 전기장 0.1 내지 10 kV/cm, 주파수 1 내지 100 Hz 및 에너지 1 내지 20 kJ로 0.1초 내지 100 초 동안 1 내지 50회 수행하는 단계;를 포함하는, 인지기능 또는 기억력 개선 효과가 증진된 브로콜리 새싹의 제조방법을 제공한다.In addition, the present invention comprises the steps of placing broccoli sprouts in a batch or continuous processing container containing a liquid medium; And performing 1 to 50 times for 0.1 to 100 seconds with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ in the liquid medium; containing, improving cognitive function or memory It provides a method for producing enhanced broccoli sprouts.
이하, 실시예에 의하여 본 발명을 상세히 설명하겠으나, 다음 실시예에 의해 본 발명이 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by examples, but the present invention is not limited by the following examples.
실시예Example
브로콜리 새싹은 (재)제주테크노파크에서 재배한 것을 구매하여 사용하였다. 고전압 펄스 전기장 처리장치는 German Institute of Food Technology사(독일)의 모델명 ELCRACK HVP5를 사용하였다.Broccoli sprouts were purchased and used at Jeju Techno Park. The high voltage pulse electric field treatment device was used as the model name ELCRACK HVP5 of German Institute of Food Technology (Germany).
실시예 1Example 1
신선한 브로콜리 새싹의 원물 그 자체를 고전압 펄스 전기장 처리장치의 물이 담긴 회분식 처리용기에 넣고 전기장 2.1 kV/cm, 주파수 100 Hz, 에너지 7 kJ 및 처리시간 3초의 처리조건에서 1회 펄스 전기장 처리하였다.The raw material of fresh broccoli sprouts itself was placed in a batch-type treatment container containing water of a high-voltage pulse electric field treatment device and treated with a pulse electric field once under the conditions of an electric field of 2.1 kV/cm, a frequency of 100 Hz, an energy of 7 kJ, and a treatment time of 3 seconds.
실시예 2Example 2
실시예 1과 동일하게 실시하되, 전기장 2.1 kV/cm, 주파수 100 Hz, 에너지 4 kJ 및 처리시간 3초의 처리조건에서 1회 펄스 전기장 처리하였다.It was carried out in the same manner as in Example 1, but was subjected to a single pulse electric field under the conditions of an electric field of 2.1 kV/cm, a frequency of 100 Hz, an energy of 4 kJ, and a treatment time of 3 seconds.
비교예 1Comparative Example 1
고전압 펄스 전기장 처리한 브로콜리와의 비교를 위해 대조군으로 신선한 브로콜리 성체를 비가식부위를 제거하고 50 g씩 절단하여 비교예 1로 하였다. For comparison with broccoli treated with a high-voltage pulsed electric field, fresh broccoli adults were removed from the unedible part as a control and cut by 50 g to obtain Comparative Example 1.
비교예 2Comparative Example 2
신선한 브로콜리 새싹을 비교예 2로 하였다. Fresh broccoli sprouts were used as Comparative Example 2.
비교예 3Comparative Example 3
실시예 1과 동일하게 실시하되, 브로콜리 새싹 대신 브로콜리 성체를 이용하여 펄스 전기장 처리하였다.It was carried out in the same manner as in Example 1, but treated with a pulse electric field using an adult broccoli instead of a broccoli sprout.
비교예 4Comparative Example 4
신선한 브로콜리 새싹에 10배 중량의 물을 첨가한 후 70 ℃에서 10 분 동안 중탕 가열하여 ESP(Epithiospecifer protein) 효소를 불활성화하였다.After 10 times the weight of water was added to fresh broccoli sprouts, the ESP (Epithiospecifer protein) enzyme was inactivated by heating in a water bath at 70° C. for 10 minutes.
이후, 상기 중탕 가열된 브로콜리 성체를 37 ℃까지 냉각시킨 후 2 시간 동안 방치하여 글루코라파닌 성분이 설포라판으로 전환되도록 미로시나제 효소 반응을 유도하였다.Thereafter, the broccoli adult body heated in the bath was cooled to 37° C. and allowed to stand for 2 hours to induce a myrosinase enzyme reaction so that the gluolapanin component was converted to sulforaphane.
비교예 5Comparative Example 5
신선한 브로콜리 성체를 비가식부위를 제거하고 50 g씩 절단한 후 상기 절단물 그 자체를 고전압 펄스 전기장 처리장치의 물이 담긴 회분식 처리용기에 넣고 전기장 2.1 kV/cm, 주파수 100 Hz, 에너지 7 kJ 및 처리시간 3초의 처리조건에서 1회 펄스 전기장 처리하였다.After removing the unedible part of fresh broccoli and cutting 50 g each, the cut itself was put into a batch-type treatment container containing water of a high-voltage pulse electric field treatment device, and an electric field of 2.1 kV/cm, a frequency of 100 Hz, and an energy of 7 kJ and One pulse electric field treatment was performed under the treatment conditions of a treatment time of 3 seconds.
그리고, 상기 펄스 전기장 처리된 브로콜리 성체에 10배 중량의 물을 첨가한 후 70 ℃에서 10 분 동안 중탕 가열하여 ESP(Epithiospecifer protein) 효소를 불활성화하였다.Then, 10 times the weight of water was added to the adult broccoli treated with the pulsed electric field, and then heated in a bath for 10 minutes at 70° C. to inactivate ESP (Epithiospecifer protein) enzyme.
이후, 상기 중탕 가열된 브로콜리 성체를 37 ℃까지 냉각시킨 후 2 시간 동안 방치하여 글루코라파닌 성분이 설포라판으로 전환되도록 미로시나제 효소 반응을 유도하였다.Thereafter, the broccoli adult body heated in the bath was cooled to 37° C. and allowed to stand for 2 hours to induce a myrosinase enzyme reaction so that the gluolapanin component was converted to sulforaphane.
비교예 6Comparative Example 6
비교예 5와 동일하게 실시하되, 브로콜리 성체 대신 브로콜리 새싹을 이용하였다. In the same manner as in Comparative Example 5, broccoli sprouts were used instead of adult broccoli.
실험예 Experimental example
실시예 1, 2 및 비교예 1 내지 6의 브로콜리를 다음의 실험에 적합한 형태로 제조하였다. 먼저, 상기 실시예 1, 2 및 비교예 1 내지 6의 브로콜리를 동결건조시켜 분말화한 후, 상기 시료들의 건조 분말 10g에 70% 메탄올 400 ml를 첨가하여 3시간 동안 교반, 추출한 추출액을 감압농축 후 동결건조시켜 농축분말을 제조하고, 이후의 실험들을 진행하였다.Broccoli of Examples 1 and 2 and Comparative Examples 1 to 6 were prepared in a form suitable for the following experiment. First, the broccoli of Examples 1 and 2 and Comparative Examples 1 to 6 were lyophilized to powder, and then 400 ml of 70% methanol was added to 10 g of the dry powder of the samples, stirred for 3 hours, and the extracted extract was concentrated under reduced pressure. After lyophilization was performed to prepare a concentrated powder, and subsequent experiments were conducted.
실험예 1: 설포라판 함량 분석Experimental Example 1: Analysis of sulforaphane content
실시예 1, 2 및 비교예 1 내지 6의 브로콜리에 포함된 설포라판 함량을 다음과 같이 분석하여 그 결과를 표 1에 나타내었다. The contents of sulforaphane contained in broccoli of Examples 1 and 2 and Comparative Examples 1 to 6 were analyzed as follows, and the results are shown in Table 1.
설포라판 표준품(90%, Sigma)으로 표준용액을 제조하여 UPLC-MS/MS(제조사 Waters)로 분석하여 표준정량선을 작성하고, 상기 시료들의 농축분말 1g을 메탄올 1 ml에 녹인 시료를 UPLC-MS/MS로 설포라판 함량을 분석하여 하기 표 1에 나타내었다. UPLC system은 Acquity UPLC system (Waters corporation, milford, MA), Binary solvent manager, Sample manager로 된 시스템이며, MS system은 Waters Quattro Premier XE Tandem MS (Micromass UK limited), Software 는 Masslynx V4.1을 이용하였다.Prepare a standard solution with sulforaphane standard (90%, Sigma) and analyze by UPLC-MS/MS (manufacturer Waters) to prepare a standard quantification curve, and a sample obtained by dissolving 1 g of the concentrated powder of the samples in 1 ml of methanol was prepared by UPLC-MS. The sulforaphane content was analyzed by /MS and is shown in Table 1 below. UPLC system is a system consisting of Acquity UPLC system (Waters corporation, milford, MA), binary solvent manager, and sample manager, MS system is Waters Quattro Premier XE Tandem MS (Micromass UK limited), software is Masslynx V4.1. .
구분division 비교예 1Comparative Example 1 비교예 2Comparative Example 2 비교예 3Comparative Example 3 비교예 4Comparative Example 4 비교예 5Comparative Example 5 비교예 6Comparative Example 6 실시예 1Example 1 실시예 2Example 2
함량(mg/g)Content (mg/g) 1.301.30 6.046.04 3.423.42 7.077.07 18.7118.71 25.4025.40 15.5915.59 15.1415.14
상대비율(%)Relative ratio (%) 100100 465465 261261 544544 14381438 19541954 11991199 11611161
상기 표 1을 살펴보면, 고전압 펄스 전기장 처리 뿐만 아니라 ESP 저해 및 미로시나제 활성화 처리를 한 비교예 5 및 비교예 6의 설포라판 함량이 가장 높은 것을 확인할 수 있다.Looking at Table 1, it can be seen that the sulforaphane content of Comparative Examples 5 and 6 subjected to the treatment of ESP inhibition and myrosinase activation as well as high voltage pulse electric field treatment was the highest.
도 1은 상기 실시예 및 비교예에 따른 브로콜리의 설포라판 함량을 더욱 잘 비교하여 나타내기 위한 HPLC 결과이고, 도 2는 상기 HPLC 결과를 그래프로 나타낸 것이다.1 is an HPLC result for better comparing the sulforaphane content of broccoli according to Examples and Comparative Examples, and FIG. 2 is a graph showing the HPLC result.
상기 도 1 및 도 2의 결과를 통해, 브로콜리 새싹을 고전압 펄스 전기장 처리 뿐만 아니라 ESP 저해 및 미로시나제 활성화 처리를 하는 경우 설포라판 함량이 가장 높은 것을 구체적으로 확인할 수 있다.From the results of FIGS. 1 and 2, it can be specifically confirmed that the broccoli sprout has the highest content of sulforaphane when the broccoli sprouts are treated with a high-voltage pulsed electric field as well as ESP inhibition and myrosinase activation treatment.
실험예 2: 동물모델에서 인지기능 또는 기억력 개선 효능 평가 - 수동회피 실험(Passive Avoidance Task)Experimental Example 2: Evaluation of the efficacy of improving cognitive function or memory in an animal model-Passive Avoidance Task
수동회피실험은 암실에서 마우스가 이전에 전기자극에 대한 기억을 기억함으로써 암실의 문으로 다시 들어가지 않는 것을 배우는 공포 동기 회피 실험 중 하나이다. 이전에 전기자극을 받은 구획으로 들어가는 것을 머뭇거리는 시간을 측정하여 측정시간을 기억능력의 지표로 삶아, 기억력을 평가하는 실험이다. The passive evasion experiment is one of the fear-motivated avoidance experiments in which the mouse learns not to enter the door of the dark room again by remembering the memory of the previous electrical stimulation in the dark room. This is an experiment that evaluates memory by measuring the time hesitated to enter the compartment previously subjected to electrical stimulation and boiling the measured time as an index of memory ability.
실험동물로는 6주령의 마우스를 ICR 마우스(26 - 28 g)를 (주)오리엔트(Seoul, Korea)에서 공급받아 동물 실험실에 약 5일간 적응시켜 사용하였으며, 물과 사료는 자유롭게 섭취하도록 하였고, 온도(23 ± 2 ℃), 습도(50 ± 10 %) 및 명암주기(12 시간)는 자동으로 조절되도록 하였다.As experimental animals, 6-week-old mice were supplied with ICR mice (26-28 g) from Orient Co., Ltd. (Seoul, Korea) for about 5 days to acclimate to an animal laboratory, and water and feed were freely ingested. The temperature (23 ± 2 ℃), humidity (50 ± 10%) and the contrast period (12 hours) were automatically adjusted.
장기투여(2주간 시료 200 mg/kg씩 매일 1회 경구투여)에 따른 그 효능을 비교하였으며 한 그룹 당 8마리씩을 사용하였다. 양성 대조군으로는 이미 기억력 개선용 치료제로 임상에서 사용되고 있는 도네페질(donepezil) 약물을 5 mg/kg 투여하였다. 스코폴라민(scopolamine) 및 도네페질(donepezil)은 Sigma-Aldrich Chemistry Co.에서 구입하여 사용하였으며, 그 외 시약은 시중에서 구입할 수 있는 최상급을 사용하였다.The efficacy according to long-term administration (200 mg/kg of sample once daily oral administration for 2 weeks) was compared, and 8 animals were used per group. As a positive control, 5 mg/kg of donepezil, which has already been used clinically as a treatment for improving memory, was administered. Scopolamine and donepezil were purchased and used from Sigma-Aldrich Chemistry Co., and other reagents were commercially available top grades.
실험시작 1시간 전에 후보 시료를 경구투여하고, 실험시작 30분 전에 scopolamine 1.2 mg/kg을 복강투여하였다. 두 약물의 효능이 극화되어 겹치는 시점에 실험을 진행하였으며, 통계처리는 Mean±SEM, ANOVA(one way analysis of variance) 및 T-test 분석법으로 하였다.The candidate sample was orally administered 1 hour before the start of the experiment, and 1.2 mg/kg of scopolamine was administered intraperitoneally 30 minutes before the start of the experiment. The experiment was conducted at the point of time when the efficacy of the two drugs was polarized and overlapped, and statistical processing was performed by means of Mean±SEM, one way analysis of variance (ANOVA) and T-test analysis.
구체적인 실험 방법으로, 스코폴라민 투여 30분 후에 쥐를 조명을 비춘 밝은 쪽 구획에 놓고 30초의 탐색시간 후 길로틴문 (guillotine door)이 열려 어두운 구획으로 들어갈 수 있게 하였다. 이때 길로틴문이 열린 후 300초 이내에 어두운 쪽으로 들어가지 않는 쥐는 실험에서 제외시켰다. 길로틴문이 열린 후 쥐가 어두운 쪽으로 들어갈 때까지의 시간을 측정하였다. 일단 쥐가 어두운 쪽으로 들어가면 길로틴문이 닫히고 0.8 mA의 전기 충격이 3초 동안 격자 바닥을 통해 흐르게 하여 이를 기억하도록 하였다(학습 시험). 학습 시험이 끝나고 24시간 후에 본 실험을 시행하였다. 쥐가 30초의 탐색시간 후 길로틴문이 열리고 어두운 쪽으로 4발이 다 들어가는데 걸리는 시간(latency time: 머무름 시간)을 300초까지 측정하였다. 걸리는 시간이 길수록 수동회피의 학습과 기억이 좋음을 나타낸다.As a specific experimental method, 30 minutes after administration of scopolamine, the rats were placed in a brightly lit compartment, and after 30 seconds of exploration time, a guilotine door was opened to allow entry into the dark compartment. At this time, mice that did not enter the dark side within 300 seconds after opening the guillotine door were excluded from the experiment. After the guillotine door was opened, the time until the rat entered the dark side was measured. Once the rat entered the dark side, the guillotine door closed and an electric shock of 0.8 mA was allowed to flow through the bottom of the grid for 3 seconds to remember it (learning test). This experiment was conducted 24 hours after the learning test was over. After the search time of 30 seconds, the time it took for the guillotine door to open and all four feet into the dark side (latency time) was measured up to 300 seconds. The longer it takes, the better the learning and memory of passive avoidance is.
도 3은 스코폴라민 유도 마우스 모델에서 실시예 및 비교예에 따른 브로콜리 시료의 경구 투여에 따른 수동회피 빈도를 통한 인지기능 또는 기억력 개선 효능을 평가하여 나타내는 그래프이다.Figure 3 is a graph showing the evaluation of cognitive function or memory improvement efficacy through passive avoidance frequency according to oral administration of broccoli samples according to Examples and Comparative Examples in a scopolamine-induced mouse model.
상기 도 3을 살펴보면, 대부분의 실험군에서 우수한 효능을 나타내었지만, 고압 펄스 전기장 처리를 한 시료(실시예)의 인지기능 또는 기억력 개선 효능이 현저히 우수한 것을 알 수 있다. 즉, 설포라판 함량이 높은 비교예 6에 비해 실시예 1의 시료의 인지기능 또는 기억력 개선 효능이 훨씬 우수한 것을 확인함으로써, 본 발명에 따른 고압 펄스 전기장 처리를 한 시료(실시예)의 경우 설포라판 이외에 인지기능 또는 기억력 개선 효과가 있는 또 다른 성분이 포함되어 있다고 추정할 수 있다.3, it can be seen that although most of the experimental groups exhibited excellent efficacy, the cognitive function or memory improvement efficacy of the sample (Example) treated with a high-pressure pulse electric field was remarkably excellent. That is, it was confirmed that the cognitive function or memory improvement effect of the sample of Example 1 was much better than that of Comparative Example 6 having a high sulforaphane content, so that the sample (Example) subjected to the high-pressure pulse electric field treatment according to the present invention was recognized other than sulforaphane. It can be assumed that it contains another ingredient that has an effect on improving function or memory.
실험예 3: 체내 아세틸콜린분해효소(acetylcholine esterase, AChE) 활성 저해 효능 측정Experimental Example 3: Inhibition of acetylcholine esterase (AChE) activity measurement in the body
기존에 인지기능 또는 기억력 개선 효과가 있다고 알려져 있는 설포라판과 실시예 및 비교예에 따른 시료들의 효능을 비교하고자 하였다. To compare the efficacy of samples according to Examples and Comparative Examples with sulforaphane, which is known to have an effect on improving cognitive function or memory.
실험동물로는 6주령의 마우스를 ICR 마우스(26 - 28 g) 를 (주)오리엔트(Seoul, Korea)에서 공급받아 동물 실험실에 약 5일간 적응시켜 사용하였으며, 물과 사료는 자유롭게 섭취하도록 하였고, 온도(23 ± 2 ℃), 습도(50 ± 10 %) 및 명암주기(12 시간)는 자동으로 조절되도록 하였다.As experimental animals, 6-week-old mice were supplied with ICR mice (26-28 g) from Orient Co., Ltd. (Seoul, Korea) and adapted to the animal laboratory for about 5 days, and water and feed were freely ingested. The temperature (23 ± 2 ℃), humidity (50 ± 10%) and the contrast period (12 hours) were automatically adjusted.
인지기능 또는 기억력 개선에 유의한 효과가 있는 설포라판의 투여량을 결정하고, 상기 결정된 설포라판 투여량을 기준으로 하여 각각 동일한 양의 설포라판을 포함하도록 하는 실시예 및 비교예에 따른 시료들의 투여량을 결정하였다. 상기 결정된 설포라판, 실시예 및 비교예에 따른 시료들의 투여량을 하기 표 2에 정리하여 나타내었다. 상기 마우스는 실험군 한 그룹 당 8마리씩을 사용하였다.Determine the dosage of sulforaphane that has a significant effect on improving cognitive function or memory, and determine the dosage of samples according to Examples and Comparative Examples to each contain the same amount of sulforaphane based on the determined sulforaphane dosage. I did. The doses of the determined sulforaphane and samples according to Examples and Comparative Examples are summarized in Table 2 below. Eight mice were used per group of the experimental group.
구분division 설포라판 1.0 mg을 포함하는 시료의 양(mg)Amount of sample containing 1.0 mg of sulforaphane (mg)
설포라판Sulforaphane 1.01.0
실시예 1Example 1 85.385.3
실시예 2Example 2 90.590.5
비교예 1Comparative Example 1 769.2769.2
비교예 2Comparative Example 2 165.5165.5
비교예 3Comparative Example 3 294.5294.5
비교예 4Comparative Example 4 141.4141.4
비교예 5Comparative Example 5 53.553.5
비교예 6Comparative Example 6 41.441.4
양성 대조군으로는 이미 기억력 개선용 치료제로 임상에서 사용되고 있는 도네페질(donepezil) 약물을 5 mg/kg 투여하였다. 스코폴라민(scopolamine) 및 도네페질(donepezil)은 Sigma-Aldrich Chemistry Co.에서 구입하여 사용하였으며, 그 외 시약은 시중에서 구입할 수 있는 최상급을 사용하였다.As a positive control, 5 mg/kg of donepezil, which has already been used clinically as a treatment for improving memory, was administered. Scopolamine and donepezil were purchased and used from Sigma-Aldrich Chemistry Co., and other reagents were commercially available top grades.
실험시작 1시간 전에 후보 시료를 경구투여하고, 실험시작 30분 전에 scopolamine 1.2 mg/kg을 복강투여하였다. 두 약물의 효능이 극화되어 겹치는 시점에 실험을 진행하였으며, 통계처리는 Mean±SEM, ANOVA(one way analysis of variance) 및 T-test 분석법으로 하였다.The candidate sample was orally administered 1 hour before the start of the experiment, and 1.2 mg/kg of scopolamine was administered intraperitoneally 30 minutes before the start of the experiment. The experiment was conducted at the point of time when the efficacy of the two drugs was polarized and overlapped, and statistical processing was performed by means of Mean±SEM, one way analysis of variance (ANOVA) and T-test analysis.
상기 시료들에 대한 아세틸콜린분해효소의 활성을 평가하기 위하여 Ellman의 방법을 이용하였다. 마우스들을 상기 설포라판, 실시예 및 비교예의 시료가 투여될 약물투여군 및 대조군 1 및 2의 11개 군(한 군당 4마리)으로 준비하였다. 상기 설포라판, 실시예 및 비교예에 따른 시료들을 약물투여군에 대하여, 상기 표 2의 결정된 용량(mg/Kg)으로 투여하였다. 한편, 대조군 1에는 PBS를 0.15 mL 투여하였고, 대조군 2에는 도네페질을 5 mg/Kg의 용량으로 투여하였다. 2주간 매일 1회 경구투여 후, 실험 동물의 뇌를 적출하여 세포분쇄(homogenation)한 뒤, 세포분쇄액을 1,000 rpm에서 10분간 원심분리한 후, 상층액에 대하여 효소 활성 평가시험을 실시하였다.Ellman's method was used to evaluate the activity of acetylcholinergic enzyme on the samples. Mice were prepared as 11 groups (4 mice per group) of the drug administration group and controls 1 and 2 to which the sulforaphane samples of Examples and Comparative Examples were administered. The samples according to the sulforaphane, Examples and Comparative Examples were administered to the drug administration group at the dose determined in Table 2 (mg/Kg). Meanwhile, 0.15 mL of PBS was administered to the control group 1, and donepezil was administered at a dose of 5 mg/Kg to the control group 2. After oral administration once daily for 2 weeks, the brains of the experimental animals were excised and homogenized, and the cell lysate was centrifuged at 1,000 rpm for 10 minutes, and then an enzyme activity evaluation test was performed on the supernatant.
뇌 내의 아세틸콜린분해효소의 활성을 평가하기 위하여, 상온에서 반응액(140 μl의 sodium phosphate buffer(pH 8.0), 40 μl의 뇌 시험용액, 10 μl의 DTNB(dithiol nitrobenzoic acid), 10 μl의 acetylcholineiodide)을 10분간 반응 시킨 후, 반응 정지 시약인 neostigmine을 이용하여 반응을 중지시켰다. 반응액을 96 well에 옮긴 후, 412 nm의 발색단에서 microplate spectrophotometer (Molecular Devices, Sunnyvale, CA, USA)을 이용하여 확인하였으며, 반응값을 식으로 계산하여 대조군 대비 효소활성도를 단위 질량당 단백질량으로 평가하였으며, 그 결과를 하기 표 3에 나타내었다. To evaluate the activity of acetylcholinease in the brain, a reaction solution (140 μl sodium phosphate buffer (pH 8.0), 40 μl brain test solution, 10 μl DTNB (dithiol nitrobenzoic acid)), 10 μl acetylcholineiodide at room temperature. ) Was reacted for 10 minutes, and then the reaction was stopped using neostigmine, a reaction stopping reagent. After transferring the reaction solution to 96 wells, it was confirmed using a microplate spectrophotometer (Molecular Devices, Sunnyvale, CA, USA) at a chromophore of 412 nm, and the reaction value was calculated by the formula, and the enzyme activity compared to the control was determined as the amount of protein per unit mass. It was evaluated, and the results are shown in Table 3 below.
구분division 아세틸콜린 분해효소의 활성 - 상대비율(%)Acetylcholine degrading enzyme activity-relative ratio (%)
대조군 1Control 1 100 ± 7.19100 ± 7.19
대조군 2(도네페질)Control 2 (Donepezil) 85.50 ± 3.2785.50 ± 3.27
설포라판Sulforaphane 78.58 ± 3.0678.58 ± 3.06
스코폴라민Scopolamine 115.03 ± 7.92115.03 ± 7.92
실시예 1Example 1 71.20 ± 1.2771.20 ± 1.27
실시예 2Example 2 72.92 ± 2.0572.92 ± 2.05
비교예 1Comparative Example 1 84.97 ± 4.2184.97 ± 4.21
비교예 2Comparative Example 2 80.97 ± 5.6180.97 ± 5.61
비교예 3Comparative Example 3 82.62 ± 10.2882.62 ± 10.28
비교예 4Comparative Example 4 77.71 ± 0.5377.71 ± 0.53
비교예 5Comparative Example 5 78.14 ± 3.4678.14 ± 3.46
비교예 6Comparative Example 6 75.09 ± 1.9275.09 ± 1.92
표 3에 나타낸 바와 같이, 설포라판이 동일한 양으로 포함되었을 경우 실시예에 따른 시료에서 아세틸콜린분해효소 저해 활성이 더욱 높다는 것을 확인할 수 있다. 특히, 설포라판에 비해 동일량의 설포라판이 포함된 실시예의 시료가 더욱 우수한 효과를 나타내는 것을 확인할 수 있다.As shown in Table 3, when sulforaphane was included in the same amount, it could be seen that the acetylcholinease inhibitory activity was higher in the sample according to the example. In particular, it can be seen that the sample of the example containing the same amount of sulforaphane exhibits a more excellent effect than that of sulforaphane.
제제예 1: 정제의 제조Formulation Example 1: Preparation of tablets
실시예 1의 브로콜리 새싹 분말 10 mg10 mg of broccoli sprout powder of Example 1
옥수수전분 100 mg100 mg corn starch
유당 100 mg100 mg lactose
스테아린산 마그네슘 2 mg2 mg of magnesium stearate
상기의 성분들을 혼합한 후 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조한다.After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet preparation method.
제제예 2: 캅셀제의 제조Formulation Example 2: Preparation of Capsule
실시예 1의 브로콜리 새싹 분말 10 mg10 mg of broccoli sprout powder of Example 1
결정성 셀룰로오스 3 mg3 mg of crystalline cellulose
락토오스 14.8 mg14.8 mg lactose
마그네슘 스테아레이트 0.2 mgMagnesium stearate 0.2 mg
통상의 캡슐제 제조방법에 따라 상기의 성분을 혼합하고 젤라틴 캡슐에 충전하여 캡슐제를 제조한다.According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.
제제예 3: 분말 건강기능식품의 제조Formulation Example 3: Preparation of powdered health functional food
실시예 1의 브로콜리 새싹 분말 1,000 mg1,000 mg of broccoli sprout powder of Example 1
비타민 혼합물 적량Vitamin mixture right amount
비타민 A 아세테이트 70 ㎍Vitamin A acetate 70 ㎍
비타민 E 1.0 mg1.0 mg of vitamin E
비타민 B1 0.13 mgVitamin B1 0.13 mg
비타민 B2 0.15 mgVitamin B2 0.15 mg
비타민 B6 0.5 mg0.5 mg of vitamin B6
비타민 B12 0.2 ㎍Vitamin B12 0.2 ㎍
비타민 C 10 mgVitamin C 10 mg
비오틴 10 ㎍Biotin 10 ㎍
니코틴산아미드 1.7 mg1.7 mg of nicotinic acid amide
엽산 50 ㎍ Folic acid 50 ㎍
판토텐산 칼슘 0.5 mg0.5 mg of calcium pantothenate
무기질 혼합물 적량Suitable amount of inorganic mixture
황산제1철 1.75 mg1.75 mg ferrous sulfate
산화아연 0.82 mgZinc oxide 0.82 mg
탄산마그네슘 25.3 mgMagnesium carbonate 25.3 mg
제1인산칼륨 15 mgPotassium monophosphate 15 mg
제2인산칼슘 55 mgDicalcium phosphate 55 mg
구연산칼륨 90 mg90 mg of potassium citrate
탄산칼슘 100 mg100 mg of calcium carbonate
염화마그네슘 24.8 mgMagnesium chloride 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강기능식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강기능식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강기능식품 조성물 제조에 사용할 수 있다.The composition ratio of the vitamin and mineral mixture is relatively suitable for health functional foods, but it is also possible to arbitrarily modify the composition, and the above ingredients are mixed according to a general health functional food manufacturing method. Then, the granules can be prepared and used for preparing a health functional food composition according to a conventional method.
제제예 4: 음료의 제조Formulation Example 4: Preparation of beverage
실시예 1의 브로콜리 새싹 분말 1,000 mg1,000 mg of broccoli sprout powder of Example 1
구연산 1,000 mg1,000 mg citric acid
올리고당 100 g100 g oligosaccharides
매실농축액 2 g2 g of plum concentrate
타우린 1 g1 g taurine
정제수를 가하여 전체 900 mLTotal 900 mL with purified water
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1 시간 동안 85 ℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 L 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 기능성 음료 조성물 제조에 사용한다. After mixing the above ingredients according to a normal health drink manufacturing method, stirring and heating the resulting solution at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and stored in a refrigerator It is used to prepare the functional beverage composition of the invention.
비록 본 발명이 상기에 언급된 바람직한 실시예로서 설명되었으나, 발명의 요지와 범위로부터 벗어남이 없이 다양한 수정이나 변형을 하는 것이 가능하다. 또한, 첨부된 특허청구범위는 본 발명의 요지에 속하는 이러한 수정이나 변형을 포함한다.Although the present invention has been described as the above-mentioned preferred embodiment, it is possible to make various modifications or variations without departing from the gist and scope of the invention. In addition, the appended claims include such modifications or variations that fall within the gist of the present invention.

Claims (15)

  1. 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 하는 인지기능 또는 기억력 개선용 건강기능식품 조성물.A health functional food composition for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
  2. 제1항에 있어서,The method of claim 1,
    상기 브로콜리 새싹은 원물, 건조물, 분쇄물, 및 추출물로 이루어진 군에서 선택된 1종 이상인 것을 특징으로 하는 건강기능식품 조성물.The broccoli sprout is a health functional food composition, characterized in that at least one selected from the group consisting of raw material, dried product, pulverized product, and extract.
  3. 제1항에 있어서, 상기 고전압 펄스 전기장 처리는 The method of claim 1, wherein the high voltage pulse electric field treatment is
    브로콜리 새싹이 담긴 액상 매질에 전기장 0.1 내지 10 kV/cm, 주파수 1 내지 100 Hz 및 에너지 1 내지 20 kJ로 0.1초 내지 100 초 동안 1 내지 50회 수행하는 것을 특징으로 하는 건강기능식품 조성물.A health functional food composition, characterized in that performing 1 to 50 times for 0.1 seconds to 100 seconds with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ in a liquid medium containing broccoli sprouts.
  4. 제1항에 있어서,The method of claim 1,
    상기 고전압 펄스 전기장 처리는 1.5 내지 2.5 kV/cm 및 에너지 3 내지 10 kJ로 수행되는 것을 특징으로 하는 건강기능식품 조성물.The high-voltage pulse electric field treatment is a health functional food composition, characterized in that carried out at 1.5 to 2.5 kV / cm and energy 3 to 10 kJ.
  5. 제3항에 있어서,The method of claim 3,
    상기 액상 매질에 담긴 브로콜리 새싹은 분쇄 또는 마쇄과정을 거치지 않은 원물 또는 그 절단물인 것을 특징으로 하는 건강기능식품 조성물.A health functional food composition, characterized in that the broccoli sprouts contained in the liquid medium are raw materials or cuts thereof that have not been pulverized or crushed.
  6. 제3항에 있어서,The method of claim 3,
    상기 액상 매질은 물, 주정 또는 이들의 혼합액인 것을 특징으로 하는 건강기능식품 조성물.The liquid medium is a health functional food composition, characterized in that water, alcohol or a mixture thereof.
  7. 제1항에 있어서,The method of claim 1,
    상기 조성물은 아세틸콜린 분해효소(acetylcholinesterase)의 활성을 저해하는 것을 특징으로 하는 건강기능식품 조성물.The composition is a health functional food composition, characterized in that inhibiting the activity of acetylcholine degrading enzyme (acetylcholinesterase).
  8. 제1항에 있어서,The method of claim 1,
    상기 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 어느 하나의 제형으로 제조되는 것을 특징으로 하는 건강기능식품 조성물.The composition is a health functional food composition, characterized in that it is prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup and beverage.
  9. 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 하는 인지기능 또는 기억력 개선용 약학 조성물.A pharmaceutical composition for improving cognitive function or memory, using broccoli sprouts treated with high voltage pulse electric field (PEF) as an active ingredient.
  10. 제9항에 있어서,The method of claim 9,
    아세틸콜린 분해효소(acetylcholinesterase) 저해 효능 및 장기적 기억력 개선 효능을 나타내는 것을 특징으로 하는 약학 조성물.A pharmaceutical composition, characterized in that it exhibits an acetylcholinesterase inhibitory effect and a long-term memory improvement effect.
  11. 인지기능 저하 개선용, 또는 인지기능 장애 예방 또는 치료용 의약 제조를 위한, 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 유효성분으로 포함하는 조성물의 용도.Use of a composition comprising broccoli sprouts treated with high voltage pulsed electric field (PEF) as an active ingredient for improving cognitive decline or for manufacturing a drug for preventing or treating cognitive function disorders.
  12. 제11항에 있어서,The method of claim 11,
    상기 인지기능 저하는 기억력 감퇴, 인지 감퇴, 식별력 감퇴 및 건망증으로 구성되는 군으로부터 선택되는 하나 이상이고, The cognitive decline is at least one selected from the group consisting of memory loss, cognitive decline, discrimination and forgetfulness,
    상기 인지기능 장애는 건망증, 알츠하이머병, 치매, 파킨슨 병, 피크병, 및 헌팅톤병으로 이루어지는 군으로부터 선택되는 퇴행성 뇌질환인 것을 특징으로 하는, 용도.The cognitive impairment is a degenerative brain disease selected from the group consisting of forgetfulness, Alzheimer's disease, dementia, Parkinson's disease, Peak's disease, and Huntington's disease.
  13. 인간, 또는 인간을 제외한 동물에게 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물을 투여하는 단계를 포함하는 인지기능 장애의 예방 또는 치료 방법.A method for preventing or treating cognitive dysfunction comprising administering a composition comprising broccoli sprouts treated with high voltage pulse electric field (PEF) to a human or non-human animal.
  14. 인간, 또는 인간을 제외한 동물에게 고전압 펄스 전기장(PEF) 처리된 브로콜리 새싹을 포함하는 조성물을 투여하는 단계를 포함하는, 개체의 인지기능 또는 기억력을 개선시키는 방법.A method for improving cognitive function or memory of an individual, comprising administering a composition comprising broccoli sprouts treated with high voltage pulsed electric field (PEF) to a human or non-human animal.
  15. 브로콜리 새싹을 액상 매질이 담긴 회분식 또는 연속식 처리 용기에 넣는 단계; 및 상기 액상 매질에 전기장 0.1 내지 10 kV/cm, 주파수 1 내지 100 Hz 및 에너지 1 내지 20 kJ로 0.1초 내지 100 초 동안 1 내지 50회 수행하는 단계;를 포함하는, 인지기능 또는 기억력 개선 효과가 증진된 브로콜리 새싹의 제조방법.Placing broccoli sprouts in a batch or continuous processing container containing a liquid medium; And performing 1 to 50 times for 0.1 second to 100 seconds with an electric field of 0.1 to 10 kV/cm, a frequency of 1 to 100 Hz, and an energy of 1 to 20 kJ in the liquid medium; containing, improving cognitive function or memory Method for producing improved broccoli sprouts.
PCT/KR2019/013692 2019-03-04 2019-10-17 Composition containing broccoli sprouts as active ingredient for improving cognitive functions or memory WO2020179982A1 (en)

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KR102301757B1 (en) * 2020-12-18 2021-09-13 서울대학교산학협력단 Natural composition suppressing a stress-induced memory loss and improving learning ability
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090073631A (en) * 2007-12-31 2009-07-03 대구한의대학교산학협력단 A composition comprising sulforaphane for preventing and treating cognitive dysfunction
KR20150011853A (en) * 2013-06-21 2015-02-03 주식회사 아이레보 Digital door lock having emergency power supply unit
KR20170023512A (en) * 2015-08-24 2017-03-06 경상대학교산학협력단 Composition for improvement of learning and memory function comprising broccoli leaf extract or its fraction as effective component

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101729913B1 (en) * 2014-04-14 2017-04-25 주식회사 비케이바이오 Preparation method for broccoli with an increased sulforaphane content and the use of it

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090073631A (en) * 2007-12-31 2009-07-03 대구한의대학교산학협력단 A composition comprising sulforaphane for preventing and treating cognitive dysfunction
KR20150011853A (en) * 2013-06-21 2015-02-03 주식회사 아이레보 Digital door lock having emergency power supply unit
KR20170023512A (en) * 2015-08-24 2017-03-06 경상대학교산학협력단 Composition for improvement of learning and memory function comprising broccoli leaf extract or its fraction as effective component

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
AKIHIRO SHIINA, NOBUHISA KANAHARA , TSUYOSHI SASAKI , YASUNORI ODA , TASUKU HASHIMOTO , TADASHI HASEGAWA , TAISUKE YOSHIDA , MASAO: "An open study of sulforaphane-rich broccoli sprout extract in patients with schizophrenia", CLINICAL PSYCHOPHARMACOLOGY AND NEUROSCIENCE, vol. 13, no. 1, 2015, pages 62 - 67, XP055737887 *
MATUSHESKI N V; JUVIK J A; JEFFERY E H: "Heating decreases epithiospecifier protein activity and increases sulforaphane formation in broccoli", PHYTOCHEMISTRY, vol. 65, no. 9, 2004, pages 1273 - 1281, XP004512269 *
SIYOUNG LEE, JISUNG KIM , SANG GWON SEO , BO-RYOUNG CHOI , JUNG-SOO HAN , KI WON LEE , JIYOUNG KIM: "Sulforaphane alleviates scopolamine-induced memory impairment in mice", PHARMACOLOGICAL RESEARCH, vol. 85, 2014, pages 23 - 32, XP055737891 *

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