KR20140006375A - Composition comprising allyl isothiocyanate for preventing and curing of angiogenesis - Google Patents

Abstract

The present invention relates to a composition containing Allyl isothiocyanate as an active ingredient for preventing and treating angiogenesis. According to the present invention, the Allyl isothiocyanate has an excellent effect in inhibiting angiogenesis, thereby being used as the active ingredient in a pharmaceutical composition and health functional food for preventing and treating angiogenesis. [Reference numerals] (AA,CC,GG,KK) Normal group; (BB,DD,HH,LL) Control group; (EE,II,MM) Experimental group 1; (FF,JJ,NN) Experimental group 2

Description

Composition for preventing and treating angiogenesis containing allyl isothiocyanate as an active ingredient {cOMPOSITION COMPRISING Allyl isothiocyanate FOR PREVENTING AND CURING OF angiogenesis}

The present invention relates to a composition for preventing and treating angiogenesis, which contains allyl isothiocyanate as an active ingredient.

The term "angiogenesis" refers to angiogenesis inhibitors developed by Dr. Judah Folkman of Harvard Medical School in early May 1998 when angiostatin and endostatin are introduced into rats. After reports of anti-cancer activity, it became widely known, and now, as one of the effective means for the treatment of cancer, the method of inhibiting neovascular formation is intensively studied.

In general, neovascularization is caused by a series of processes involving angiogenic factors, and in normal processes such as embryonic growth, reproduction, development, and wound repair. The control is performed smoothly and occurs, but if the control mechanism is abnormal, diabetic retinopathy, neovascular glaucoma, rheumatoid arthritis, kaposi's sarcoma, psoriasis, chronic inflammation serious diseases such as chronicinflamation and hemangiomas.

In this regard, vascular endothelial growth factor (VEGF) has been identified as a key factor involved in stimulating angiogenesis and inducing vascular permeability (Ferrara et al., Endocr. Rev. 18: 4-25, 1997). The characteristics of the rat VEGF gene were confirmed and its expression pattern in the embryogenesis was analyzed. Continuous expression of VEGF was observed in epithelial cells adjacent to the window-shaped endothelium, for example, choroid and kidney glomeruli. This data is consistent with the role of VEGF as a multifunctional regulator of endothelial cell growth and differentiation (Breier, G. et al., Development, 1992, 114, 521). VEGF correlates with angiogenesis in vascular tissues (eg lung, heart, placenta and solid tumors) (Binetruy-Tourniere et al., EMBO J. 2000, 19, 1525). For example, VEGF is involved in promoting solid tumor growth and metastasis by stimulating tumor-associated angiogenesis (Lu et al., J. Biol. Chem. 2003, 278, 43496). On the other hand, since the expression of VEGF is essential for restoration of vascular tissue in particular, VEGF has been proposed to be used to enhance vascular tissue repair. Indeed, VEGF showed the disadvantage that, like other proteins in animals, the molecule was so large that it could not cross the vascular barrier and had a short half-life, resulting in a short-term dissipation. In recent years, studies have been actively conducted to find out important genes involved in vascularization, specifically acting on vascular endothelial cells, and to utilize these genes to treat various diseases requiring angiogenesis. However, There are no significant results.

On the other hand, white blood cells in the human body circulate in the human body to effectively recognize and remove antigens, and when an emergency situation such as antigen invasion or tissue damage occurs, it moves to such a place to remove antigens. You can move directly through the walls of blood vessels to the target site, this series of processes are called inflammatory reactions. Inflammatory reactions can effectively remove antigens, but excessive or persistent reactions can damage surrounding tissues and cause disease.

Regarding techniques for inhibiting angiogenesis or inflammation, there are Korean Patent Registration No. 10-0758006, Korean Patent Publication No. 10-2009-0057074, and Korean Patent Publication No. 10-2003-7005651. The present inventors have discovered the angiogenesis or inflammation inhibitory effect of allyl isothiocyanate to arrive at the present invention.

It is an object of the present invention to provide a substance having angiogenic efficacy and a composition containing the same as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition having an inhibitory activity against angiogenesis containing allyl isothiocyanate as an active ingredient. In the present invention, the pharmaceutical composition may be characterized by inhibiting the proliferation of endothelial cells, may be characterized by inhibiting the formation of endothelial tubes, it may be characterized by inhibiting the invasion of endothelial cells. In addition, the pharmaceutical composition may be characterized in that it further comprises a pharmaceutically acceptable additive, the pharmaceutical composition may be to have a formulation of tablets, capsules, solutions, suspensions, emulsions or syrups.

Furthermore, the present invention provides a health functional food for preventing and improving angiogenesis containing allyl isothiocyanate as an active ingredient. In the present invention, the pharmaceutical composition may be characterized by inhibiting the proliferation of endothelial cells, may be characterized by inhibiting the formation of endothelial tubes, it may be characterized by inhibiting the invasion of endothelial cells. The health functional food may be in the form of powder, granules, tablets, capsules or beverages.

In the present invention, allyl isothiocyanate (AITC) is a spice component such as black mustard and wasabi, and exists as a thio glycoside in plants, and adds cell destruction or water (melting wasabi and mustard). In this case, the isothiocyanate is released by the action of the enzyme, and the chemical formula is the same as the following Chemical Formula 1.

[Formula 1]

The composition of the present invention, the allyl isothiocyanate 0.1 to 50% by weight. The composition comprising allyl isothiocyanate of the present invention may further comprise a suitable carrier, excipient or diluent according to conventional methods. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The compositions according to the invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories or sterile injectable solutions, respectively, according to conventional methods. have. More specifically, when formulating the composition, it can be prepared using a diluent or an excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, allyl isothiocyanate, It can be prepared by mixing sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. in addition to commonly used diluents such as water and liquid paraffin . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The present invention may be varied depending on the age, sex and body weight of the patient, but it is generally administered in an amount of 0.01 to 500 mg / kg, preferably 0.1 to 100 mg / kg, once to several times per day . The dosage may also be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections. Since allyl isothiocyanate of the present invention has little toxicity and side effects, it can be used safely even for prolonged administration for prophylactic purposes.

The present invention provides a health functional food comprising the allyl isothiocyanate and food acceptable food supplement additives, including various foods, for example, beverages, gums, teas, vitamin complexes, health supplements, and the like. , Pills, powders, granules, acupuncture, tablets, capsules or beverages. At this time, the amount of the allyl isothiocyanate in the food or beverage can be generally added at 0.01 to 15% by weight of the total food weight in the case of the health food composition of the present invention, 0.02 based on 100 ml in the case of the health beverage composition To 10 g, preferably 0.3 to 1 g.

Food supplementary additives as defined herein include food additives customary in the art, such as flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, and the like. The health beverage composition according to the present invention is an essential ingredient in the ratio indicated, and there is no particular limitation on the ingredients added in addition to the allyl isothiocyanate and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Can be. Examples of the natural carbohydrate include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; Polysaccharides such as dextrin, cyclodextrins; And sugar alcohols such as xylitol, sorbitol, and erythritol. As natural flavors other than those described above, natural flavors (such as tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and synthetic flavors (saccharin, aspartame, etc.) have. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

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.

As described above, the allyl isothiocyanate according to the present invention has excellent anti-angiogenic and anti-inflammatory effects, so it can be used as a pharmaceutical composition and health functional food for prevention and treatment of angiogenesis containing it as an active ingredient. Can be.

1 is a graph showing the effect on the weight, degree of disease, long length of allyl isothiocyanate.
Figure 2 shows the effect on the anatomical change of allyl isothiocyanate.
3 is a graph showing the effect on the MPO activity of allyl isothiocyanate.
Figure 4 shows the effect on the angiogenesis of allyl isothiocyanate.
Figure 5 shows the effect of allyl isothiocyanate on VEGF-A, VEGFR-2.
Fig. 6 shows the effect of allyl isothiocyanate on iNOS and Cox-2.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Example  1. Confirmation of Inflammatory Inhibitory Effect of Allyl Isothiocyanate

1-1. Enteritis  Inhibitory Effects in the Model

To study angiogenesis and inflammation inhibition by allyl isothiocyanate (AITC), experiments were conducted by inducing inflammatory bowel disease with dextran sodium sulfate (DSS) in a c57bL / 6 mouse model. It was. Allyl isothiocyanate was purchased from Sigma, and the chemical formula is shown in the following Chemical Formula 1.

[Formula 1]

Forty-eight males of c57bL / 6 8-week males were purchased and randomly divided into four groups. And the mouse model was dieted under the conditions of Table 1 below.

group Dietary Way Normal group No treatment Control group Dextran sodium sulfate (Inflammatory bowel disease induction group) Experiment 1 Dextran Sodium Sulfate + Allyl Isothiocyanate 10uM (D + AITC10) Experiment 2 Dextran Sodium Sulfate + Allyl Isothiocyanate 25uM (D + AITC 25)

First, allyl isothiocyanate was pretreated to the oral cavity for a week and then dissolved in 3% DSS in water instead of water. Allyl isothiocyanate was continuously administered while feeding dextran sulfate.

As a result of daily weight, stool status, and blood severity for a week, the weight, disease acitivity index (DAI) and colon length were significantly decreased by DSS. However, in the group treated with allyl isothiocyanate, it was confirmed that the Disease acitivity index (DAI) was significantly recovered (FIG. 1). Since DAI is measured based on weight, stool status, blood level, etc., DAI may be a more important factor.

1-2. Enteritis  Anatomical Changes in the Model

Sodium dextran sulfate is known to irritate the intestinal mucosa and cause inflammatory reactions and damage tissues. In order to observe the effect of allyl isothiocyanate on tissues damaged by dextran sulfate, inflammation severity and extension of inflammation were observed to examine its anatomical findings through H / E staining. injury, and the degree of bowel gland injury was scored.

As a result, the dextran sodium sulfate treated group was severely damaged, the inflammation continued to the mucosa, and the intestinal gland and epithelial cells were severely damaged. However, in the group to which allyl isothiocyanate was administered, these phenomena decreased significantly depending on the concentration (FIG. 2).

1-3. In enteroinflammatory model  Effect on Inhibition of Inflammation

Enteritis is known to activate various inflammatory cells. Dextran sodium neutrophil infiltration was investigated by measuring Myeloma peroxide (MPO) activity. As a result, it was observed that MPO activity increased by sodium dextran sulfate was significantly reduced depending on the concentration by allyl isothiocyanate (FIG. 3).

1-4. In enteroinflammatory model iNOS , Cox Effect on -2

Inflammatory agents iNOS, Cox-2 are known to increase in enteritis. Western blot was performed to see if allyl isothiocyanate can affect their expression.

As a result, it was observed that the expression of NOS and Cox-2 increased by dextran sodium sulfate significantly decreased in Experimental Group 1 and Experimental Group 2. In addition, a significant decrease was observed in VEGFR-2, a receptor of VEGF-A (FIG. 6).

Example  2. Confirmation of Inhibition of Angiogenesis of Allyl Isothiocyanate

2-1. In enteroinflammatory model  Effect on angiogenesis

Chronic enteritis is known to cause extensive tissue damage, remodeling, inflammatory cell infiltration, loss of epithelial integrity, and angiogenesis. The purpose of this study was to investigate the effect of allyl isothiocyanate on increased angiogenesis in enteritis caused by sodium dextran sulfate. As a result, it was observed that the blood vessel density increased significantly in the group treated with dextran sodium sulfate. However, a significant decrease was observed in the group to which allyl isothiocyanate was administered (FIG. 5).

2-2. In enteroinflammatory model VEGF -A, VEGFR Effect on -2

Angiogenesis, which causes tissue damage in chronic inflammatory bowel injury, is known to increase the expression of VEGF-A. This increased expression of VEGF-A increases the expression of adhesion molecules, enhances leukocyte adhesion, increases the infiltration of inflammatory cells and increases tissue damage. In addition, VEGF-A increases vascular permeability and is known to inhibit the growth of vascular endothelial cells required for vascular stabilization, proliferation of vascular endothelial cells, and vascular leakage. In order to investigate whether allyl isothiocyanate may affect the expression of VEGF in enteritis caused by dextran sodium sulfate, western blot was performed.

As a result, it was observed that the expression of VEGF-A increased by dextran sodium sulfate was significantly decreased in Experimental Group 1 and Experimental Group 2. In addition, the expression of VEGFR-2, a receptor of VEGF-A, was increased by sodium dextran sulfate as in VEGF-A. (FIG. 6).

Examples of formulations for the composition of the present invention are illustrated below. However, the following formulation examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following formulation examples.

Formulation example  One: Sanje  Produce

Allyl isothiocyanate 300 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation example  2: Preparation of tablets

Allyl Isothiocyanate 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation example  3: Preparation of capsules

Allyl Isothiocyanate 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4: Preparation of injection

Allyl Isothiocyanate 50 mg

Appropriate sterile distilled water for injection

pH adjuster

(2 ml) per 1 ampoule according to the usual injection preparation method.

Formulation example  5: Liquid  Produce

Allyl Isothiocyanate 100 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Formulation example  6: Manufacture of health food

Allyl isothiocyanate 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation example  7: Manufacture of health drinks

Allyl isothiocyanate 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, ≪ / RTI >

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

While the present invention has been described with reference to exemplary embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. You will know. In addition, many modifications may be made to adapt a particular situation and material to the teachings of the invention without departing from the essential scope thereof. Accordingly, it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention, but that the invention be construed as including all embodiments falling within the scope of the appended claims.

Claims (11)

A pharmaceutical composition having inhibitory activity against angiogenesis, containing allyl isothiocyanate as an active ingredient. The method of claim 1,
The pharmaceutical composition is a pharmaceutical composition having an inhibitory activity against angiogenesis, characterized in that it inhibits the proliferation of endothelial cells. The method of claim 1,
The pharmaceutical composition has an inhibitory activity against angiogenesis, characterized in that to inhibit the formation of endothelial cells. The method of claim 1,
The pharmaceutical composition is a pharmaceutical composition having an inhibitory activity against angiogenesis, characterized by inhibiting the invasion of endothelial cells. The method of claim 1,
Wherein the pharmaceutical composition further comprises a pharmaceutically acceptable additive. The method of claim 1,
Wherein the pharmaceutical composition has a formulation of a tablet, a capsule, a solution, a suspension, an emulsion or a syrup. Health functional food for preventing and improving angiogenesis, containing allyl isothiocyanate as an active ingredient. 8. The method of claim 7,
The health functional food is a health functional food for preventing and improving angiogenesis, characterized in that to inhibit the proliferation of endothelial cells. 8. The method of claim 7,
The health functional food is a health functional food for preventing and improving angiogenesis, characterized in that to inhibit the formation of endothelial cells. 8. The method of claim 7,
The health functional food is a health functional food for preventing and improving angiogenesis, characterized in that to inhibit the invasion of endothelial cells. 8. The method of claim 7,
Health functional food for preventing and improving angiogenesis, characterized in that the powder, granules, tablets, capsules or beverage form.

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