KR20070089288A - Composition comprising an extract of ammomum xanthioides wallich showing antioxidative effect - Google Patents

Abstract

A composition comprising an extract of Amomum xanthioides Wallich is provided to be usefully used as a pharmaceutical composition and a health functional food for preventing and treating diabetic complications such as stroke, myocardial infarction and diabetic ocular disorders by showing excellent free-radical scavenging activity and lipid peroxidation inhibition effect and decreasing the oxidative stress. The anti-oxidative composition comprises 0.001-50 wt.% of an extract of Amomum xanthioides Wallich as an effective ingredient. The extract is prepared by using a polar solvent selected from the group consisting of water, C1-4 lower alcohol and a mixture thereof. The health functional food comprises the extract and is in the form of tablet, capsule, powder, granule, liquid phase or pill.

Description

Composition containing an extract of Ammomum xanthioides Wallich showing antioxidative effect

1 is a diagram showing the antioxidant activity of the sine extract measured by hydrogen donating ability by DPPH (1,1-diphenyl picrylhydrazyl) method,

Figure 2 is a diagram measuring the content of TBARS (thiobarbituric acid-reactive substances) in the diabetic animal model liver tissue.

The present invention relates to a composition containing a sinus extract having antioxidant activity as an active ingredient.

The major causes of death in Korea in 2003 were cancer, cerebrovascular disease, cardiovascular disease, and diabetes mellitus (Korea National Statistical Office: The cause of death Statistics 2003. Annual Report of on the Cause of Death Statistics.Korea National Statistical Office, 2004 ). Reactive oxygen species (ROS) may be a major factor causing chronic degenerative diseases such as cardiovascular disease, diabetes mellitus, cancer, degenerative neuropathy and aging, and antioxidants are known to help prevent chronic degenerative diseases. (Bray, TM Nutrition. 16 , pp 578-581, 2000). Since all aerobic organisms inevitably carry harmful free radical species during metabolism, all organisms must have a mechanism to remove these free radical species in order to survive. Active oxygen species can be broadly classified into two types: compounds that form oxygen radical species, such as hydrogen peroxide, and oxygen radical species. As is already known, hydrogen peroxide is converted to oxygen or water by catalase or peroxidase, but in the case of oxygen radical species removal mechanism, oxygen radicals other than O _2- (superoxide anion) are removed. Species removal mechanisms are not yet clear.

Diabetes complications are known to be caused by elevated blood glucose levels and free radicals caused by diabetes (Kazuyuki Hayashi, Masakazu Haneda, Daisuke Koya et al. Diabetes Res. Clin. Pract., 52 , pp85-96, 2001: Joshua P Klein and Stephen G Waxman.The Lancet Neurology , 2 (9) , pp548-554, 2003). In the diabetic state, the production of free radicals is increased by hyperglycemia such as glucose autooxidation and non-enzymatic protein glycosylation, which is very susceptible to oxidative damage caused by various free radicals. The Korean Diabetes Association ( 1998, Korea Medicine). Hydrogen peroxide produced under normal conditions can be removed by intracellular antioxidant systems and antioxidant enzymes, but lipid peroxidation is increased in abnormal conditions such as diabetes (Wills ED Biochem J. , 99 , pp667-675, 1965). Shweta Bhatiaa, et al., Clinical Biochemistry , 36 , pp 557-562, 2003). Sustained hyperglycemia during diabetes can increase the production of oxygen free radicals through autooxidation and glycosylation of non-enzymatic proteins, and oxygen free radicals result in lipid peroxidation of membrane phospholipids, which increases the lipid oxidation product MDA.

In patients with diabetes, free radicals are one of the major mechanisms involved in the development of chronic complications. Diabetes patients are prone to free radical production, which leads to increased levels of lipid peroxidation in liver tissue, heart muscle and serum, leading to cardiovascular diseases such as stroke and myocardial infarction (Joshua P Klein and Stephen G Waxman, The Lancet Neurology , 2 (9) , pp 548-554, 2003). In diabetic patients, the incidence of vascular diseases such as coronary heart disease and atherosclerosis is known to be 2 to 6 times higher than that of non-diabetic patients. Cardiac circulatory complications are the leading cause of death in diabetic patients. In addition, diabetic eyes, like other organs, are susceptible to metabolic disorders caused by diabetes. Increased non-enzymatic glycosylation, oxidation of proteins and lipids, and reduced glutathione in diabetes are harmful substances. It reduces the skin's resistance to and causes clouding and cataracts of the lens (Yarat A., et al., Free Rad. Biol. Med., 31 (9) , pp 1038-1042, 2001).

TBARS content, which indicates an increase in plasma lipid peroxidation, was higher in diabetic patients with cardiovascular complications than in patients without complications (MM Kesavulu, et al., Diabetes Research and Clinical Practice , 53 (1) , pp33-39, 2001 ). SOD (superoxide dismutase) is an enzyme that decomposes free radicals into hydrogen peroxide and is one of the detoxification systems in vivo, and reports a positive correlation between SOD activity and lifespan in animal liver (De Mulder CLC, et al., J . Nutr. Biochem., 6, pp452, 1995) shows a good anti-oxidizing of SOD. SOD is a metalloprotein, and metals such as Cu, Zn, Mn, and Fe are present in its active site, and among them, Cu, ZN-SOD and Mn-SOD are found in animal tissues.

Sign of the invention ( Ammomum xanthioides Wallich ) is a perennial herb belonging to the Zingiberaceae family. It is distributed in southern China, Southeast Asia and India. The medicinal form is oval or oval, with three dull corners, 15-20 mm long and 10-15 mm in diameter. In agreement, it is written that the stomach is sore and bulge that it is used when vomiting and nausea rather than going down. Known ingredients include essential oils of seeds, the main components of which are Borneol, Bornyl-acetate, Linalool, and Nerolidol. Literature Review on Origin and Form, Korean Journal of Herbology, pp. 96-105, 1986). However, scientific and systematic studies and results indicating that ingestion of autographs has an effect of preventing or improving antioxidant and diabetic complications are insufficient.

Accordingly, the present inventors prepared a sine having excellent antioxidant activity, and measured the free radical scavenging activity and the concentration of lipid peroxide, and completed the present invention by confirming that the lipid peroxidation inhibitory effect was excellent in preventing diabetes complications. It was.

It is an object of the present invention to provide a composition and a dietary supplement containing the signature extract having antioxidant activity as an active ingredient.

In order to achieve the above object, the present invention provides an antioxidant composition as an active ingredient extract of Ammomum xanthioides Wallich having antioxidant activity.

The extract includes an extract available in a polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably ethanol.

Antioxidant composition containing the extract of the present invention contains 0.1 to 50% by weight of the signature extract relative to the total weight of the composition.

Hereinafter, the present invention will be described in detail.

After freeze-drying and crushing the sine of the present invention, a C ₁ to C ₄ lower alcohol such as water, methanol, ethanol, butanol, or the like, containing about 2 to 20 times, preferably about 5 to 15 times the volume of the powder sample, or A mixed solvent having a mixing ratio of about 1: 0.1 to 1:10, preferably hot water extraction, cold extraction, reflux cooling weight, and ultrasonic extraction for about 1 to 24 hours, preferably 6 to 15 hours at room temperature by adding methanol. Extraction method such as, preferably extracted by cold extraction method, and then filtered under reduced pressure to separate the extract and the residue, and the concentrated residue is concentrated under reduced pressure by repeating the process for the filtered residue several times to obtain the extract of the present invention. .

In addition, conventional fractionation processes can also be carried out (Harborne JB Phytochemical methods: A guide to modern techniques of plant analysis , 3rd Ed., Pp 6-7, 1998).

Sign extract obtained by the above method significantly reduces the activity of lipid peroxide TBARS and increases the activity of antioxidant enzymes SOD (superoxide dismutase) and catalase, diabetic complications caused by oxidative stress Prevents and treats

In addition, sine is a medicine that has been used for a long time or edible herbal extracts of the present invention extracted therefrom also have no problems such as toxicity and side effects.

The composition comprising the extract of the present invention may further include appropriate carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the sign extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.

The composition comprising the sign extract according to the present invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to a conventional method. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the signature include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in the signature. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the cause extract of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the preferred effect, the sign extract of the present invention is preferably administered at 0.0001 to 100mg / kg, preferably 0.001 to 100mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

Sign extract of the present invention can be administered to various mammals such as rats, mice, livestock, humans. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The present invention provides a health functional food containing a sign extract having antioxidant activity as an active ingredient.

The dietary supplement of the present invention comprises the extract in an amount of 0.01 to 95%, preferably 1 to 80% by weight, based on the total weight of the composition.

In addition, it is possible to manufacture and process as a health functional food in the form of tablets, capsules, powders, granules, liquids, pills and the like for the purpose of antioxidant effects.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the signature as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the sign extract of the present invention is a flavor, such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and Salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the cause of the present invention may contain a pulp for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components can 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 sines of the present invention.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Experimental Examples.

Example 1. Preparation of Sine Extract

1-1. Sine methanol extract manufacturer

After freeze-dried and crushed sine purchased from Gwangmyeong medicinal herbs, 100% methanol corresponding to 10 times the weight of the sine sample (100g) was added and extracted using a magnetic stirrer for 12 hours. The extract was separated from the residue by filtration under reduced pressure, and 100% methanol corresponding to 5 times the weight of the sinusoid at the time of extraction was added to the obtained residue, followed by extraction using a magnetic stirrer for 6 hours. Again, the resultant was filtered under reduced pressure, and the residue was extracted. 100% methanol, which is three times the weight of our signature, was added and extracted using a magnetic stirrer for three hours, followed by filtration and concentration under reduced pressure to obtain 5 g of the final signature extract. .

1-2. Cod Ethanol Extract Manufacturer

In the same manner as in Example 1-1, 5 g of a final autograph extract was obtained using 95% ethanol as a solvent.

Reference Example 1. Experiment Preparation

Corn starch used in this experiment was purchased from the subject company, casein was purchased from Murry goulburn Co. Australia, sucrose and choline bitartrate was purchased from Samyang. α-cellulose was purchased from Sigma, Louis, MO, USA, mineral mixtures, vitamin mixtures and L-cystine were purchased from ICN biochemical (USA), soybean oil was purchased from Cheil Sugar. .

Experimental Example 1. Measurement of DPPH free radical scavenging activity

1-1. Experiment preparation

A: (Antioxidant determination by the use of a stable free radical, Nature, 26, 1199-1200, 1958 Blois, MS) DPPH method (1,1-diphenyl picrylhydrazyl) for measuring antioxidant activity in vitro (In vitro) of a sine Was used.

The DPPH method is reduced by physiologically active substances that exhibit antioxidant activities such as tocopherol, ascorbate, flavonoid compounds, aromatic amines, Maillard browning products, and peptides. It is a method to measure the antioxidant effect by hydrogen donating ability according to the degree of dark purple discoloration. At this time, L-ascorbic acid was used as a control, and 2 x 10 ^-4 M DPPH was added to each sample of the concentration of the sine extract obtained in Example 1, and the absorbance was measured at 517 nm. The sample concentration was 0.5 mg / mL (see Table I).

1-2. Experiment result

To determine the IC ₅₀ value, the sample concentration was diluted to 0.25 mg / mL, 0.1 mg / mL, 0.05 mg / mL, 0.025 mg / mL, and the scavenging ability of 60.9%, 49.3%, 31.7%, and 13.4%, respectively. Indicated. Therefore, the sign extract inhibited the enzyme activity in a concentration-dependent manner, IC ₅₀ value was 0.11mg / mL showed a high free radical scavenging ability (see Figure 1). In other words, it is known that cell damage caused by free radicals is closely related to the development of diabetes mellitus and diabetes complications. Therefore, it is confirmed that sine active ingredient exhibiting strong antioxidant effect can improve diabetic complications. Can be.

Free radical scavenging ability according to concentration (%) Concentration (mg / ml) L-ascorbic acid Sine extract 0.5 97.4 68.2

Experimental Example  2. Diabetic Rat  Determination of Hypoglycemic Effect of Caesar Extract

2-1. Experiment preparation

In order to measure the effect of improving the in vivo diabetic complications of sine extract, which was shown to have excellent in vitro antioxidant activity in Experimental Example 1, 14 db / db mice of type 2 diabetic model were treated for 1 week. After the adaptation period, diets containing AIN-93G basal diet (see Table 2 below) were added to the control group and 10% lyophilized sign was used for the control group. Was taken for 7 weeks. The diet and drinking water were allowed to be ad libitum freely. The temperature and humidity of the feeding room were maintained at 20 ~ 25 ℃ and 50 ~ 60%, respectively, and the lighting was turned on and off every 12 hours. Animal weight and dietary intake were measured three times a week.

Dietary composition of the control group Furtherance Control Diet (%) Cornstarch 39.7 Casein 20.0 Dextrinized starch 13.2 Sucrose 10.0 α-cellulose 5.0 Mineral mixtures 3.5 Vitamin mixtures 1.0 L-cystine 0.3 Choline Bitartrate 0.25 Tert-Butylhydroquinone 0.0014 Soybean oil 7.0

Animals were sacrificed at 6 weeks of diet, fasted for 14 hours before sacrifice, suffocated with carbon dioxide gas, and blood was collected from the heart with a syringe containing 10 mg of EDTA (ehylene diamine tetra acetic acid). Blood was centrifuged at 3,000 rpm for 15 minutes to collect plasma and stored at -70 ° C. Liver tissue was extracted and stored at -70 ° C.

All results were expressed as mean ± standard deviation, and the significance test between the control group and the sign extract administration group was performed using the Student's t-test (α = 0.05).

2-2. Experiment result

As a result of measuring body weight and dietary intake, the control group and the sign group weighed 42.1 ± 3.4 g and 40.7 ± 3.0 g, respectively. 4.7 ± 0.5g / day, 4.9 ± 0.6g / day, and dietary efficiency was not significantly different between the control group and the cause of death (10.2 ± 1.1%, 9.4 ± 0.9%, respectively) (see Table 3 below).

Body weight and dietary intake of control and autographs in diabetic animal models Control Autograph Body weight (g) ¹⁾ 42.1 ± 3.4 ²⁾ 40.7 ± 3.0 Dietary Intake (g / day) 4.7 ± 0.5 4.9 ± 0.6 Dietary Efficiency (%) ³⁾ 10.2 ± 1.1 9.4 ± 0.9

¹⁾ Weight after 6 weeks of dietary intake

²⁾ Mean ± SD

³⁾ weight gain (g) / dietary intake (g)

Experimental Example 3 Measurement of TBARS Content in Liver

3-1. Experiment preparation

The measurement of liver TBARS (thiobarbituric acid-reactive substances) content was measured using thiobarbituric acid (Ohkawa H., Ohisi N., and Yagi K. Anal Biochem , 95 , pp351, 1979). The content of malondialdehyde (MDA) reacting with TBA) was measured, and 1,1,3,3, -tetramethoxypropane (TMP) was used as a control. In addition, the protein content of the homogeneous solution was measured using the modified Lowry's method (Marklund S. and Marklund G. Eur J Biochem , 47 , p 469, 1974). TBARS content in the liver is shown in FIG. 2 .

3-2. Experiment result

The TBARS content (1.9 ± 0.4 nmol MDA / mg protein) of liver tissue in the autologous group was significantly decreased (P <0.05) compared to the control (2.5 ± 0.3 nmol MDA / mg protein). In other words, the intake of the cause extract may inhibit the lipid peroxidation of the liver and reduce the oxidative stress of the diabetic rats, thereby contributing to the prevention of tissue damage and diabetic complications caused by diabetes.

Experimental Example 4. Acute Toxicity Test

Acute toxicity test was performed using 6-week-old specific pathogen-free (SPF) SD rats. Two animals per group were orally administered to the animals of the present invention at a dose of 100 mg / kg. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed, and hematological and hematological examinations were performed.

As a result of this experiment, there were no clinical symptoms or dead animals in all animals treated with the test substance, and no toxicity change was observed in weight change, blood test, blood biochemical test and autopsy findings. As a result, the cause of death of the present invention did not show a toxicity change in rats up to 100 mg / kg, respectively, and the minimum lethal dose (LD ₅₀ ) was determined to be a safe substance of 100 mg / kg or more.

Hereinafter, an example of the preparation of a pharmaceutical composition containing the extract of the present invention will be described, but the present invention is not intended to limit the present invention but is only intended to be described in detail.

Formulation Example 1 Preparation of Powder

Cod Extract 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

Cod Extract 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Cod Extract 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

Cod Extract 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

_{Na 2 HPO 4 · 12H 2 O} 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Cod Extract 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, lemon flavor is added to the mixture, and then the above ingredients are mixed, purified water is added to adjust the total amount to 100 ml, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Health Food

Cod Extract 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

Vitamin B ₁₂ 0.2 μg

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

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 90mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

Cod Extract 100 mg

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

0.25 g of vitamin B ₁

0.3 g of vitamin B ₂

Water quantification

After mixing the above components according to a conventional healthy beverage production method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container sealed sterilization and then refrigerated and stored in the present invention For the manufacture of healthy beverage compositions.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

As described above, the sign extract of the present invention is excellent in free radical scavenging activity, excellent lipid peroxidation inhibitory effect, by reducing the oxidative stress cardiovascular system such as stroke, myocardial infarction caused by oxidative stress in diabetic patients It can be used for medicines and functional foods for the prevention and treatment of complications caused by diabetes such as diseases, diabetic eye diseases.

Claims (5)

Antioxidant composition containing an extract of Ammomum xanthioides Wallich as an active ingredient. The composition of claim 1, wherein the extract is an extract available in a polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The composition of claim 1, wherein the extract comprises 0.001 to 50% weight percent based on the total weight of the composition. Sine with Antioxidant Effect ( Ammomum xanthioides Wallich ) Health functional food containing extract as an active ingredient. The dietary supplement according to claim 4 in the form of tablets, capsules, powders, granules, liquids, pills.

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