KR101404566B1 - Composition for preventing or treating dementia comprising Tenebrio molitor extract - Google Patents

Abstract

The present invention relates to a composition for preventing or treating dementia comprising a Tenebrio molitor extract, which comprises a pharmaceutical composition for preventing or treating dementia comprising brown gruel extract as an active ingredient, a food composition for preventing or improving dementia will be. Since the brown gruel extract of the present invention has an effect of inhibiting β-secretase (BACE1) activity, it is effective for the pharmaceutical composition for the prevention or treatment of dementia or for the preparation of the food composition for preventing or improving dementia.

Description

[0001] The present invention relates to a composition for preventing or treating dementia comprising a brown goat extract,

The invention mealworm (Tenebrio The present invention relates to a composition for preventing or treating dementia, which comprises an extract of brown gruel as an active ingredient, and a food composition for preventing or alleviating dementia.

Due to the rapid increase of the elderly population in modern society, senile dementia is emerging as a serious social problem. Despite the high incidence of this disease, there is currently no clear remedy or preventative agent, which is leading to massive socioeconomic losses. Alzheimer's disease (AD) is the most common form of dementia and is now the third leading cause of death in the United States following cardiovascular disease and cancer. Alzheimer's disease is a progressive neurodegenerative disease characterized by memory loss and cognitive impairment. Alzheimer's disease is characterized by the accumulation of plaques in nerve tissue and blood vessels, entanglement of nerve fibers, amyloid deposition, synaptic damage and nerve cell death.

Tacrine (Cognex, 1994) and donepezil (Aricept, 1996) have been approved by the US FDA as a representative dementia remedy known so far. The mechanism of action of these drugs is to inhibit the activity of acetylcholinesterase (AChE) enzyme, which is a degradative enzyme of acetylcholine which plays a central role in the central nervous system, to increase the concentration of acetylcholine, a neurotransmitter, It is known to cure. However, the tacrine is more expensive than the efficacy and has a serious problem of hepatotoxicity. Although the donepezil is not toxic to the liver, it stimulates the parasympathetic nerve and causes various side effects such as vomiting, nausea and diarrhea. In addition, the drugs are not medicines for therapeutic treatment such as improvement of cerebral lesions, but are merely drugs for alleviating major symptoms of dementia such as memory loss.

Therefore, researches for developing a new type of dementia treatment agent which can cause treatment without any side effects as described above are being actively carried out. One of them is trying to develop a substance that can block the toxicity of ß-amyloid, which is known to be the causative agent of Alzheimer's disease.

B-secretase (BACE 1) is one of the target enzymes for the development of new dementia therapies with a clear mechanism of action and no side effects. Alzheimer's disease is caused by an abnormality of an amyloid gene existing on human chromosome 21, which is known to be mainly due to a genotoxic substance called β-amyloid (Probst A. et al., Brain Pathol., 1: 229-239, 1991). Amyloid is produced by decomposing amyloid precursor protein (APP) present in normal brain into C-terminal peptide and β-amyloid by β-secretase (BACE 1) do.

The β-amyloid and C-terminal peptides are present in small amounts in normal brain, but they are known to be produced in large amounts in the brain of Alzheimer's patients to form senile plaques (Kang, J., Nature, 325: 733-736, 1987). Amyloid and C-terminal peptides by inhibiting the increase of [beta] -amyloid and C-terminal peptides by degradation of amyloid precursor protein (APP) when inhibiting the activity of [beta] -secretase Can inhibit senile neuroprotection and amyloid gene abnormality, resulting in the treatment of dementia.

The development of inhibitors for conventional BACE 1 is based mostly on synthetic compounds and BACE 1 inhibitors derived from natural products are relatively minor.

The brown duck (Tenebrio molitor) is an insect of the beetle neck and is also called a brown rice duck. The body length is about 15mm, and the body color is similar to Kowloon-ku, dark brown and glossy. There are dots on the head but not densely. The tip of tactile sense is the same length and butterfly. The back of the breast is square, the new groove of the rear rim is distinct, and the middle is a pentagram with a wide bow. The wing wings are brown, with eight new grooves, the convex side of the liver, and small dots. Adult larvae lay eggs mainly in grains, eggs are so small that they can not be easily found by the eyes and hatch after 12 weeks. Extracts of brown goats are called mealworms, and they are largely reared on food insects and pets. The duration of the extract is about 10 weeks, but about two weeks for artificial rearing. The reason that wheat worms are used for pets is that they are relatively clean insects because they have relatively short periods of transformation, and they are very clean insects that feed on grain. The extract is molted 920 times depending on the prey and temperature until it becomes a pupa. It becomes a pupa and becomes an adult after 23 weeks. When it first becomes allegory, it changes to light brown or gradually black. Adults are nocturnal, hiding in the hole during the daytime and active at night. It grows in pupa and adultery in spring when it is wintering with extract in the grain bag near the main shrine. It is famous for grain pests and has been growing in recent zoos as food for small animals such as birds and hedgehogs, and is distributed throughout Korea and around the world.

Currently, the patent using brown gurus includes a brown gurr-containing feed (Patent Publication No. 10-2011-0054692), an antifungal protein isolated from a brown gurr extract (Korean Patent No. 10-0230959) .

Thus, the present inventors have found that a brown gutter extract inhibits the activity of BACE1, thereby completing the present invention.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating dementia comprising an extract of Tenebrio molitor as an active ingredient.

Another object of the present invention is to provide a medicament containing the pharmaceutical composition as an active ingredient.

Another object of the present invention is to provide a food composition for preventing or alleviating dementia comprising an extract of Tenebrio molitor as an active ingredient.

Another object of the present invention is to provide a health functional food comprising the food composition as an active ingredient.

In order to achieve the object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating dementia comprising an extract of Tenebrio molitor as an active ingredient.

According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the pharmaceutical composition as an active ingredient.

In order to accomplish another object of the present invention, the present invention provides a food composition for preventing or alleviating dementia comprising an extract of Tenebrio molitor as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a health functional food comprising the food composition as an active ingredient.

Hereinafter, the present invention will be described in detail.

The invention mealworm (Tenebrio The present invention also provides a pharmaceutical composition for preventing or treating dementia comprising an extract of Lactobacillus molitor as an active ingredient.

The present invention also provides a medicament comprising the pharmaceutical composition as an active ingredient.

The brown gruel extract of the present invention comprises (a) a brown gruel ( Tenebrio molitor ) powder with an organic solvent having 1 to 6 carbon atoms; And (b) fractionating the extract obtained in the step (a) with one or more solvents selected from the group consisting of hexane, dichloromethane, ethyl acetate, butanol and water, and concentrating under reduced pressure. .

The organic solvent having 1 to 6 carbon atoms in the step (a) may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, is selected from the group consisting of benzene, chloroform, ethyl acetate, methylene chloride, hexane, and cyclohexane.

In the step (b), the extract obtained in step (a) is fractionally extracted with at least one solvent selected from the group consisting of hexane, dichloromethane, ethyl acetate, butanol, and water and concentrated under reduced pressure. In the fraction extraction, most preferably, the substances listed above can be sequentially fractionated.

In the present invention, 10-fold (w / v) ethanol was added to the brown gruel powder at the highest growth stage. To the concentrated ethanol extract, 500 mL of water and the same amount of hexane were added, and the hexane fraction was obtained by layer separation with a fractionating funnel. Then, dichloromethane, ethyl acetate, butanol and water fractions were obtained in the same manner, and the obtained extracts were subjected to concentration under reduced pressure (see Example 1).

As a result of measuring the inhibitory effect of the extracts on the β-secretase (BACE1) activity, hexane, dichloromethane, ethyl acetate, butanol and water extracts showed inhibitory activity (see Experimental Example 1). Beta-secretase is also referred to as BACE1 (-site APP-cleaving enzyme 1) or mepsin 2 (memapsin 2). Some studies to date have shown that overexpression of the transmembranous aspartic acid protease BACE increases the activity of beta-secretase and overexpression of BACE causes APP to cleave the beta-secretase , The activity of beta-secretase is inhibited by the BACE antisense oligonucleotide, and the purified BACE cleaves APP precisely to show beta-secretase properties (Robert Vassar et al., Secretase Cleavage of Alzheimer's Amyloid Precursor Protein by the Transmembrane Aspartic Protease BACE, Science, 286, 735-741 (1999)) and the membrane-binding protein aspartic acid protease also inhibit the formation of beta-secretase beta-amyloid (Riqiang Yan et al., Membrane-anchored aspartyl protease with Alzheimer's disease-secrease activity, Nature, 402, 533-537 (1999)) There godoego.

In particular, beta-secretase inhibits beta-amyloid activity by inhibiting beta-secretase activity as an enzyme that initiates beta-amyloid production, and also prevents all subsequent harmful steps, - Modifying the gene coding for the secretase does not have serious problems and is therefore clinically safe and has many advantages as a target for the treatment of Alzheimer's disease. Accordingly, many inhibitors for inhibiting the activity of beta-secretase have been reported.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating dementia comprising a brownish goat extract as an active ingredient, and a food composition for preventing or alleviating dementia. Dementia diseases for which preventive, therapeutic or ameliorative effects of the pharmaceutical compositions and food compositions of the present invention are not limited include but are not limited to Alzheimer type dementia, cerebrovascular dementia, senile dementia and dementia caused by head injury.

The pharmaceutical composition according to the present invention may contain, alone or in combination with one or more pharmaceutically acceptable carriers, excipients or diluents, a brown gruel extract or a pharmaceutically acceptable salt thereof.

The pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. In addition, the carrier for parenteral administration may contain water, a suitable oil, a saline solution, an aqueous glucose and a glycol, and may further contain a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, and the like in addition to the above components. Other pharmaceutically acceptable carriers can be found in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, Pa., 1995).

The composition of the present invention can be administered to mammals including humans by any method. For example, it can be administered orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal administration Lt; / RTI >

The pharmaceutical composition of the present invention can be formulated into oral preparations or parenteral administration preparations according to the administration route as described above.

In the case of a preparation for oral administration, the composition of the present invention may be formulated into a powder, a granule, a tablet, a pill, a sugar, a tablet, a liquid, a gel, a syrup, a slurry, . For example, an oral preparation can be obtained by combining the active ingredient with a solid excipient, then milling it, adding suitable auxiliaries, and then processing the mixture into a granular mixture. Examples of suitable excipients include, but are not limited to, sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, and starches including corn starch, wheat starch, rice starch and potato starch, Cellulose such as methylcellulose, sodium carboxymethylcellulose and hydroxypropylmethyl-cellulose and the like, fillers such as gelatin, polyvinylpyrrolidone and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may optionally be added as a disintegrant. Further, the pharmaceutical composition of the present invention may further comprise an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent.

In the case of a preparation for parenteral administration, it can be formulated by a method known in the art in the form of injection, cream, lotion, external ointment, oil, moisturizer, gel, aerosol and nasal aspirate. These formulations are described in Remington's Pharmaceutical Science, 15th edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a commonly known formulary for all pharmaceutical chemistries.

The total effective amount of the brown gruel extract of the present invention or a pharmaceutically acceptable salt thereof may be administered to a patient in a single dose and may be administered in a fractionated treatment protocol ). ≪ / RTI > In the pharmaceutical composition of the present invention, the content of the active ingredient may be varied depending on the degree of the disease. Preferably, the preferred total dose of the brown gruel extract of the present invention or a pharmaceutically acceptable salt thereof may be about 0.01 to 1,000 mg, most preferably 0.1 to 100 mg per patient body weight per day.

However, the amount of the brown gruel extract or its pharmaceutically acceptable salt may be varied depending on various factors such as the route of administration and the number of treatments of the pharmaceutical composition as well as the age, weight, health, sex, severity of disease, diet and excretion rate of the patient A person skilled in the art will appreciate that the brown gurder derivative or a pharmaceutically acceptable salt thereof is suitable for the prophylactic or therapeutic treatment of dementia, An effective dose can be determined. The pharmaceutical composition according to the present invention is not particularly limited to the formulation, administration route and administration method as long as the effect of the present invention is exhibited.

The food composition according to the present invention may be prepared in the form of a composition by mixing with known active ingredients known to have a preventive or ameliorative effect on dementia diseases, and may further include a food-acceptable food supplementary additive. The food composition of the present invention includes compositions for all types of food such as functional foods, nutritional supplements, health foods and food additives. Food compositions of this type may be prepared in a variety of forms according to conventional methods known in the art. For example, the health food may be in the form of a gum, a vitamin complex, a tea, a juice or a drink containing a brown gruel extract. The food composition containing the brown gruel extract as an active ingredient may be granulated, encapsulated or powdered It can be ingested. The functional food composition of the present invention may contain components that are ordinarily added in food production, and may include, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, in the case of being made with a drink, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, jujube extract, licorice extract, etc. may be further added in addition to the brown gruel extract of the present invention.

The food supplementary additives defined in the present invention include food additives customary in the art, such as flavors, flavors, colorants, fillers, stabilizers and the like, and are exemplified below.

In addition to the above-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like.

In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

 The present invention also provides a health functional food comprising, as an active ingredient, a food composition for preventing or improving gum disease comprising the brown gurrhoid extract as an active ingredient.

As the health food, the lactic acid bacteria or fermented lactic acid bacteria of the present invention may be prepared in the form of tea, juice, and drink, and may be ingested, granulated, encapsulated and powdered. In addition, the lactic acid bacteria or the fermented lactic acid bacteria of the present invention may be mixed with other active ingredients to prepare a composition.

Since the brown gruel extract of the present invention has an effect of inhibiting β-secretase (BACE1) activity, it is effective for the pharmaceutical composition for the prevention or treatment of dementia or for the preparation of the food composition for preventing or improving dementia.

FIG. 1 is a graph showing the activity of β-secretase (BACE1) inhibition of brown goat extract. (Hex: hexane fraction extract, EA: dichloromethane, ethyl acetate fraction extract, BuOH: butanol fraction extract, H ₂ O: water fraction extract)

Hereinafter, the present invention will be described in detail.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

< Example  1>

Manufacture of brown goat extract

Powder of brown frog (28 ~ 35mm in length, 5 ~ 7mm in diameter) was obtained from World Way Co., Ltd. 1.2 kg of brown gruel powder was stirred in 12 liters of ethanol for 7 days at room temperature at 250 rpm and then concentrated under reduced pressure to obtain 380 g of ethanol extract.

The ethanol extract (380 g) was dissolved in 500 mL of water, and the solution was added to a fractionating funnel. Hexane (hexane) equivalent to water was added thereto, followed by shaking at room temperature, followed by layer separation to obtain hexane fractions. Thereafter, the reaction mixture was fractionated in succession with a solvent of a different polarity such as CH ₂ Cl ₂ (dichloromethane), EtOAc (ethyl acetate), n-BuOH and H ₂ O, and then concentrated under reduced pressure (40 rpm, 100 rpm) Fractions were obtained. Hexane fraction _{313.90g (82.61%), CH 2} Cl 2 fractions 0.42g (0.11%), EtOAc fractions 0.78g (0.21%), BuOH fraction 24.80 g (6.53%), H 2 O fraction 39.03g (10.27%) a .

Thin layered chromatography (TLC) of the fractions confirmed that there was no difference in the component patterns of the CH ₂ Cl ₂ and EtOAc (ethyl acetate) fractions, and the two fractions were mixed and named EtOAc.

<Experimental Example 1>

ß- secretase  Measurement of active inhibition effect

BACE1 FRET assay kit (PANVERA) was used to measure β-secretase (BACE1) inhibitory activity of the fractions prepared in Example 1. The positive control group was 'resveratol' (Sigma, St. Louis, USA) known as BACE1 inhibitor. Louis, MO, USA). Resveratrol is known to have about 50% BACE1 inhibitory activity at 15 uM.

10 μl of 750 nM BACE1 substrate (Rh-EVNLDAEFK-Quencher), 10 μl of BACE1 (1.0 U / ml) and 50 μg / ml of the fractions of Example 1 with assay buffer (50 mM sodium acetate, pH 4.5) 10 μl of each sample diluted to 100 μg / mL was placed in a 384-well plate and reacted at 25 ° C. for 60 minutes. Fluorescence was measured at Ex: 530 nm and Em: 590 nm using a fluorescence microplate reader, The inhibition rate was calculated by the following formula.

Inhibition (%) = [1 - ( S - S ₀ ) / ( C - C ₀ )] 100

C : Fluorescence of control after 60 min of incubation (enzyme, assay buffer and substrate)

C ₀ : Fluorescence of the control at time 0

S : Fluorescence of control after 60 min of incubation (enzyme, sample solution and substrate)

S ₀ : Fluorescence of the tested samples ( S ) at time 0

The experimental results are shown in Fig. In Fig. 1, it can be confirmed that all fractions have inhibitory activity, and it can be seen that the higher the concentration, the higher the inhibition ability.

Since the brown gruel extract of the present invention has an effect of inhibiting the activity of β-secretase (BACE1), a pharmaceutical composition for preventing or treating dementia, or a food composition for preventing or alleviating dementia can be produced, which is highly industrially applicable.

Claims (8)

A pharmaceutical composition for preventing or treating dementia comprising an extract of Tenebrio molitor as an active ingredient.
The method of claim 1, wherein the extract
(a) extracting a powder of Tenebrio molitor with an organic solvent having 1 to 6 carbon atoms; And
(b) fractionating the extract obtained in step (a) with one or more solvents selected from the group consisting of hexane, dichloromethane, ethyl acetate, butanol and water, and concentrating under reduced pressure .
The method according to claim 2, wherein the organic solvent having 1 to 6 carbon atoms in step (a) is selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, Wherein the composition is selected from the group consisting of ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, and cyclohexane.
The composition according to claim 1, wherein the dementia is at least one disease selected from the group consisting of Alzheimer type dementia, cerebrovascular dementia, senile dementia and dementia caused by head injury.
A composition for preventing or improving dementia comprising an extract of Tenebrio molitor as an active ingredient.
The composition according to claim 5, wherein the dementia is at least one disease selected from the group consisting of Alzheimer type dementia, cerebrovascular dementia, senile dementia and dementia caused by head injury.
A medicament containing the composition of claim 1 as an active ingredient.
A health functional food comprising the composition of claim 5 as an active ingredient.

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