KR20150113687A - A composition for prevention or treatment of osteoporosis comprising extract of Allium hookeri - Google Patents

Abstract

The present invention relates to a composition for preventing and treating/alleviating osteoporosis, which contains an Allium hookeri extract as an active ingredient. The Allium hookeri extract of the present invention increases expression of osteoblast differentiation adjustment genes (ALP and ColA1) to exhibit excellent effects of preventing and treating osteoporosis. Accordingly, the composition is a natural material, thereby being safely used without side effects. Osteoblast differentiation genes are adjusted, so the composition exhibits an excellent effect of preventing and treating/alleviating osteoporosis.

Description

TECHNICAL FIELD The present invention relates to a composition for prevention or treatment of osteoporosis,

The present invention relates to a composition for preventing or treating osteoporosis, which comprises Allium hookeri extract as an active ingredient, and more particularly to a composition for preventing and treating osteoporosis, which comprises a tea extract as an active ingredient.

Osteoporosis is the most common skeletal disease among bone related diseases such as multiple myelosis and osteoarthritis, and is generally characterized by an increase in fracture and a decrease in bone strength. For normal bone function, bone remodeling by bone homeostasis of bone formation by osteoclast and osteoblast is needed (for example, Rev. Endocrinol 8: 212227, 2011, J. Dent. Res. 91: 736-744, 2012).

However, excessive activity of osteoclasts and deactivation of osteoblasts cause imbalance in the remodeling process, leading to adult skeletal diseases such as osteoporosis. In order to alleviate this imbalance, methods for suppressing excessive activity of osteoclasts, promoting osteoclast activity, inhibiting osteoclast activity and promoting osteoclast activity are generally used, and they are used for treatment of osteoporosis (Nat. Rev. Rheumatol. 7: 631-638, 2011).

Osteoblasts are formed from mesenchymal stem cells. The mineralization, including calcium, formed by osteoblast differentiation not only maintains bone strength but also plays an important role in the homeostasis of calcium and hormone metabolism throughout the body. Calcium formation by osteoblast differentiation is regulated by vitamin D and parathyroid hormone, and it is known that bone morphogenetic protein (BMP), Wnt, MAP kinase, calcineurin-calmodulin kinase Alkaline phosphatase (ALP), which is involved in the differentiation of osteoblasts, has been synthesized at the early stage of differentiation by the cross-talk of various signaling systems such as calcineurin-calmodulin kinase, NF-B and AP- (Osteopontin; OPN), osteocalcin (OCN), type I collagen (ColA1), etc. related to mineralization are synthesized, thereby forming bone by osteoblast differentiation (Nat Rev. Drug Discov. 11: 234-250, 2012).

 A representative treatment for osteoporosis developed so far is bisphosphonates, which inhibits bone loss by weakening the functions of osteoclasts (bone resorption-related cells). Merck's Fosamax, Procter Gamble And "Actonel" of the company. However, these treatments do not promote the formation of bones, and as a decomposition-inhibiting agent, it is necessary to substitute bone because it can induce additional fractures in continuous use, so it can not be a fundamental treatment. (Datamonitor Research Reports, 2007).

Allium hookeri is called Root Leek. In Korea, it is called "三 菜" because it has sweetness, bitter taste, and pungent flavor. Sanjay is native to the Himalayas above 1,400 meters above sea level and is distributed in China, India, Bhutan, Sri Lanka and Myanmar. Myanmar is a private prescription for various inflammatory and cancer diseases and is being used as edible and medicinal. Protein, sugar, fiber, ascorbic acid, phytosterol, total phenol, etc. are contained more than onions, and dietary sulfur compounds are known to be 6 times more than garlic. Inhibition of DPPH radical scavenging activity and nitric oxide (NO) production by anti-inflammatory effects in lipopolysaccharide-induced macrophages (J. Korean Soc. Food Sci. Nutr. 41: 1645-1648, 2012) Soc. Food Sci. Nutr. 42: 1351-1356, 2013). However, prior to the present invention, there has been no known effect on the prevention and treatment of osteoporosis of tubercle bark extract.

The present inventors have conducted a study to explore the natural substances that can be applied has a promoting effect on activity of osteoblasts safely, three (Allium hookeri extract of the present invention has an effect of promoting osteoblast activity, thus completing the present invention.

It is an object of the present invention to provide a composition for preventing or treating osteoporosis, which comprises a tubercle leaf extract as an active ingredient.

In order to achieve the above object, the present invention provides a composition for preventing or treating osteoporosis, which comprises Allium hookeri extract as an active ingredient.

Hereinafter, the contents of the present invention will be described in more detail.

The present invention provides a composition for the prevention and treatment of osteoporosis, which comprises a tubular extract as an active ingredient.

The present invention is not limited to a method for producing a tubular extract, but is preferably a method for producing a tubular extract from water or a part (leaf or root) of a tubular plant or a plant, an organic solvent having 1 to 6 carbon atoms and a subcritical or supercritical fluid Or more. It can also be prepared, if necessary, in addition to filtration and concentration steps according to methods known to those skilled in the art. In addition, specific fractions enriched in active material can be prepared using techniques such as solvent fractionation, silica gel chromatography, and prep-HPLC.

The organic solvent having a carbon number of 1 to 6 is not limited thereto, but preferably an alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl (ethyl) acetate, methylene chloride, hexane, cyclohexane, and petroleum ether.

Preferably, the tubular extract of the present invention can be obtained by extracting and drying the dried triangle using purified water, ethanol and subcritical carbon dioxide suitable for food processing, or extracting and purifying it using an ultra-high pressure extraction apparatus Or can be obtained by separating and purifying the oil obtained by directly pressing the triple plant.

The triple extract of the present invention is superior in osteoporosis effect by increasing the expression of osteoblast differentiation genes (ALP and ColA1).

In one embodiment of the present invention, an extract was prepared using a root and leaf (see Example 1).

In another embodiment of the present invention, it was confirmed that the tubercle extract increased the expression of osteoblast differentiation genes (ALP and ColA1) in MC3T3-E1 osteoblast (see Example 2).

Accordingly, the present invention provides a composition for the prevention and treatment of osteoporosis, which comprises a tubercle leaf extract as an active ingredient, and the composition of the present invention can be applied as a pharmaceutical composition or a food composition.

The tubular extract of the present invention may be used as such or in a pharmaceutically acceptable form. The term " pharmaceutically acceptable " as used herein means physiologically acceptable and does not generally cause an allergic reaction or a similar reaction when administered to humans, and the salt includes a pharmaceutically acceptable free acid, Lt; / RTI > is preferred. As the free acid, an organic acid and an inorganic acid can be used. The organic acids include, but are not limited to, citric, acetic, lactic, tartaric, maleic, fumaric, formic, propionic, oxalic, trifluroacetic, benzoic, gluconic, methosulfonic, glycolic, succinic, Glutamic acid and aspartic acid. The inorganic acid includes, but is not limited to, hydrochloric acid, bromic acid, sulfuric acid, and phosphoric acid.

The pharmaceutically effective amount of the tangerine extract according to the present invention is 0.001 to 10 mg / day / kg of body weight, preferably 0.01 to 0 mg / day / kg of body weight. However, the pharmaceutically effective amount may be appropriately changed depending on various factors such as the disease and its severity, the patient's age, weight, health, sex, administration route, and treatment period.

The term "pharmaceutically acceptable" as used herein means a non-toxic composition which is physiologically acceptable and which, when administered to humans, does not inhibit the action of the active ingredient and does not normally cause an allergic reaction such as gastrointestinal disorder, dizziness, . Such carriers include all kinds of solvents, dispersion media, oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads and microsomes.

Meanwhile, the pharmaceutical composition according to the present invention may be formulated together with a suitable carrier according to the route of administration. The administration route of the pharmaceutical composition according to the present invention is not limited thereto, but it can be administered orally or parenterally. Parenteral routes of administration include, for example, various routes such as transdermal, nasal, peritoneal, muscular, subcutaneous or intravenous.

In addition, when administered parenterally, the pharmaceutical composition of the present invention can be formulated together with a suitable parenteral carrier according to methods known in the art in the form of injection, transdermal drug delivery, and nasal inhalation. In the case of such injections, they must be sterilized and protected against contamination of microorganisms such as bacteria and fungi. Examples of suitable carriers for injectables include, but are not limited to, solvents or dispersion media containing water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and / or vegetable oils . More preferably, suitable carriers include isotonic solutions such as Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose Etc. may be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal and the like may be further included. In addition, the injections may in most cases additionally include isotonic agents, such as sugars or sodium chloride.

Examples of transdermal dosage forms include ointments, creams, lotions, gels, solutions for external use, pastes, liniments, and air lozenges. By "transdermal administration" as used herein, it is meant that the pharmaceutical composition is locally administered to the skin so that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin. These formulations are described in Remington's Pharmaceutical Science, 15th Edition, 1975, Mack Publishing Company, Easton, Pennsylvania, which is a commonly known formulary in pharmaceutical chemistry.

In the case of inhalation dosage forms, the compounds used according to the present invention may be formulated into a pressurized pack or a pressurized pack using a suitable propellant, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. It can be conveniently delivered in the form of an aerosol spray from a nebulizer. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges for use in an inhaler or insufflator may be formulated to contain a compound, and a powder mixture of a suitable powder base such as lactose or starch.

Other pharmaceutically acceptable carriers can be found in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, Pa., 1995).

The pharmaceutical composition according to the present invention may also contain one or more buffers (e.g., saline or PBS), a carbohydrate (e.g., glucose, mannose, sucrose or dextran), an antioxidant, a bacteriostat, (E. G., EDTA or glutathione), an adjuvant (e. G., Aluminum hydroxide), a suspending agent, a thickening agent and / or a preservative.

In addition, the pharmaceutical compositions of the present invention can be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal.

In addition, the pharmaceutical composition of the present invention may be administered in combination with a known compound having an effect of preventing or treating osteoporosis.

Further, the tubular extract according to the present invention may be provided in the form of a food composition for the purpose of preventing or improving osteoporosis. The food composition of the present invention includes all forms of functional foods, nutritional supplements, health foods, food additives and feeds, It is targeted for eating. Food compositions of this type may be prepared in a variety of forms according to conventional methods known in the art.

For example, as a health food, the tubular extract of the present invention itself can be prepared in the form of tea, juice, and drink and then consumed for drinking, granulated, encapsulated, and powdered. In addition, the triple extract of the present invention can be prepared in the form of a composition by mixing together with known active ingredients known to be effective for preventing and improving osteoporosis.

Functional foods also include beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruits, bottled, jam, maalmalade, etc.), fish, meat and processed foods such as ham, Etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, Protein, retort food, frozen food, various kinds of seasoning (for example, soybean paste, soy sauce, sauce, etc.).

In order to use the tubular extract of the present invention in the form of a food additive, it may be used in the form of powder or concentrate.

The preferred content of the tubular extract of the present invention in the food composition of the present invention may include about 0.01 to 100% by weight based on the total weight of the food.

INDUSTRIAL APPLICABILITY As described above, the present invention provides a composition for preventing or treating osteoporosis, the composition comprising a tubular extract as an active ingredient. The triple extract of the present invention shows an excellent effect on osteoporosis by increasing the expression of osteoblast differentiation genes (ALP and ColA1). Accordingly, the present invention provides a composition which can be safely used as a natural product without side effects, and exhibits an excellent effect for prevention and treatment / improvement of osteoporosis.

FIG. 1 shows mRNA expression levels of osteoblast differentiation genes (ALP and ColA1) in MC3T3-E1 osteoblast by treatment with triplex extract.

Hereinafter, the present invention will be described in detail by way of examples.

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

Triptych  Preparation of extract

<1-1> Preparation of ethanol extract of Samgyet roots

The dried triangular root was pulverized with a mixer, and 100 g of the pulverized triple root sample was added to 1 L of ethanol and extracted with stirring at 50 for 60 minutes. The extracted sample was filtered with filter paper of Whatman No. 2, and the filtered extract was concentrated with a vacuum rotary condenser to remove solvent components to obtain a triple rooted ethanol extract.

<1-2> Preparation of hot water extract of Samchab root

The dried triangular root was pulverized with a mixer, and 100 g of the pulverized triangular root sample was added to 1 L of water and extracted with stirring at 100 for 4 hours. The extracted samples were filtered with Wattmann filter paper No. 2, and the filtered extract was concentrated with a vacuum rotary condenser to remove the solvent components, thereby obtaining a watery extract of Samchab root.

&Lt; 1-3 > Preparation of Methanol Extract of Samjip Root

The dried triangular root was pulverized with a mixer, and 100 g of the pulverized triple root sample was added to 1 L of methanol and extracted with stirring at 50 for 60 minutes. The extracted sample was filtered with Wattman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary condenser to remove the solvent component to obtain a methanol extract of Trichotomous root.

&Lt; 1-4 > Preparation of triple rooted hexane extract

The dried triangular root was pulverized with a mixer, and 100 g of the pulverized triple root sample was added to 1 L of nucleic acid and extracted with stirring at 50 for 60 minutes. The extracted sample was filtered with a filter paper of Wattman No. 2, and the filtered extract was concentrated with a vacuum rotary condenser to remove solvent components to obtain a triple rooted hexane extract.

&Lt; 1-5 > Preparation of triacylglycerol ethyl acetate extract

The dried triangular root was pulverized with a mixer, and 100 g of the pulverized triangular root sample was added to 1 L of ethyl acetate and extracted with stirring at 50 for 60 minutes. The extracted sample was filtered with a filter paper No. 2 of Wattman, and the filtered extract was concentrated with a vacuum rotary condenser to remove solvent components to obtain a triple root ethyl acetate extract.

<1-6> Preparation of chloroform-extracted triplex root

The dried rootstock root root was ground with a mixer, and 100 g of the ground root root root sample was added to 1 L of chloroform and extracted with stirring at 50 for 60 minutes. The extracted sample was filtered with Wattman No. 2 filter paper, and the filtered extract was concentrated with a vacuum rotary condenser to remove solvent components to obtain a triple rooted chloroform extract.

<1-7> Preparation of Ethanol Extract of Triangular Leaves

The dried triangular leaf was ground with a mixer, and 100 g of the pulverized triangular leaf sample was added to 1 L of ethanol and extracted with stirring at 50 for 60 minutes. The extracted sample was filtered through filter paper of Whatman No. 2, and the filtered extract was concentrated with a vacuum rotary condenser to remove solvent components to obtain a triangular leaf ethanol extract.

MC3T3 - E1  In osteoblasts Triptych  Effect of extract treatment on osteoblast differentiation

MC3T3-E1 osteoblast cells (ATCC, Manassas, Va., USA) were cultured in α-MEM (α-modified Eagle's Medium) medium containing 10% fetal bovine serum, And placed in a 6-well microtiter plate at a rate of 1,00,000 cells / well. L-ascorbic acid (Sigma, St. Louis, Mo., USA) and 10 mM -glycerol phosphate disodium salt hydrate (Sigma, St. Louis, Mo., USA) at the density of 100% ), The oocyte ethanol extract prepared in Example 1-1 was treated at a concentration of 10, 20 ppm to induce osteoblast differentiation for 3 days. RT-PCR was performed to examine the mRNA expression levels of the major genes ALP and ColA1 involved in osteoclast differentiation. Total RNA was harvested from the differentiated cells using TRIzol reagent (Invitrogen, Carlsbad, Calif., USA) and reverse transcribed. RT-PCR analysis was performed as follows. First, the RNA was reverse transcribed using reverse transcriptase for cDNA synthesis. RT-PCR was performed with the following specific primers:

β-actin

Forward primer: 5'-AGCCATGTACGTAGCCATCC-3 '(SEQ ID NO: 1)

Reverse primer: 5'-CTCTCAGCTGTGGTGGTGAA-3 '(SEQ ID NO: 2)

ALP

Forward primer: 5'-CCATGGTAGATTACGCTCACA-3 '(SEQ ID NO: 1)

Reverse primer: 5'-ATGGAGGATTCCAGATACAGG-3 '(SEQ ID NO: 2)

ColA1

Forward primer: 5'-ACTCAGCCGTCTGTGCCTCA-3 '(SEQ ID NO: 1)

Reverse primer: 5'-GGAGGCCTCGGTGGACATTA-3 '(SEQ ID NO: 2)

As a result, as shown in Fig. 1, it can be confirmed that the extract of Sanchi extract increases the amount of mRNA expression of ALP and ColA1, the osteoblast differentiation genes. Therefore, it was confirmed that osseed extract treatment promoted bone formation in MC3T3-E1 osteoblasts.

INDUSTRIAL APPLICABILITY As described above, the present invention provides a composition for preventing and treating osteoporosis, which comprises a tubular extract as an active ingredient. The triple extract of the present invention has an excellent prophylactic and therapeutic effect on osteoporosis by increasing expression of osteoblast differentiation control genes (ALP and ColA1). Therefore, the present invention can be safely used as a natural product without side effects, and it is possible to provide a composition showing an excellent effect for prevention and treatment / improvement of osteoporosis by controlling osteoblast differentiation gene, and thus it is highly likely to be used industrially.

Claims (4)

Allium hookeri ) extract as an active ingredient for preventing or ameliorating osteoporosis.
A pharmaceutical composition for the prevention and treatment of osteoporosis, comprising a tubular extract as an active ingredient.
The composition according to claim 1 or 2, wherein the tubular extract is an extract of at least one solvent selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and a subcritical or supercritical fluid.
4. The method according to claim 3, wherein the organic solvent having 1 to 6 carbon atoms is selected from the group consisting of alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate Wherein the composition is selected from the group consisting of methylene chloride, hexane, cyclohexane, and petroleum ether.

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