KR20150139025A - Moringa extracts having anti-oxidant and anti-diabetic activity - Google Patents

Abstract

The present invention relates to a Moringa extract having a radical scavenging activity, an antioxidant activity, and a hypoglycemic activity. The present invention can also use the antioxidant and anti-diabetic activities of the Moringa extract to provide an antioxidant composition, a hypoglycemic composition, a dietary supplement product for prevention and alleviation of diabetes and diabetic complications, and a pharmaceutical composition for the prevention and alleviation of diabetes and diabetic complications. All of the compositions can contain the Moringa extract as an active ingredient. According to the present invention, the dietary supplement product and the pharmaceutical composition can manage the blood sugar level of a patient and prevent diabetes in case of normal users.

Description

[0002] MORINGA EXTRACTS HAVING ANTI-OXIDANT AND ANTI-DIABETIC ACTIVITY WITH ANTIOXIDANT AND ANTI-

The present invention relates to a Moringa extract having radical scavenging activity, antioxidative activity and hypoglycemic action.

Currently, about 3% of the world's population, 150 million people suffer from diabetes and complications, and by 2030 there will be about 300 million people who are suddenly surged by lack of exercise, bad eating habits and over-consumption of high energy foods . According to the statistics of the health authorities in Korea recently, the number of diabetes patients in Korea is 8.3 per 100 people, reaching 4 million, and the number of new cases is estimated to be 500,000 per year. If the current trend continues, it will exceed 14 people per 100 people by 2030. In addition, as the average life span of Koreans increases, the pace of progress toward an aging society is accelerating, and the number of patients with senile diabetes and complications is rapidly increasing.

Diabetes is a chronic metabolic disease in which glucose is excreted in the urine while insulin is not effectively used due to deficiency or insufficiency of insulin produced in the pancreas or abnormal function of insulin due to abnormal function of glucose due to decrease of the biomolecule using glucose . More seriously, when the hyperglycemic state is left untreated, three major complications such as retinopathy, kidney failure, and atherosclerosis can occur, resulting in damage to the whole blood vessels and reaching a fatal condition.

More specifically, hyperglycemia is caused by decreased insulin secretion or insulin resistance of peripheral tissues due to β-cell destruction of the pancreas due to genetic, metabolic, and environmental factors. In particular, the production of lipid peroxide in vivo is further accelerated This leads to excessive cellular production of free radicals (O2-, HO-), which leads to cellular deterioration such as protein destruction, chromosome aberration, and red cell destruction, and cell necrosis. Diabetes is caused and promoted by the abnormal increase of free radicals, and antioxidants are involved in the signal transduction of specific genes to maintain normal blood sugar. For free radicals, the biotissues are composed of endogenous removers such as superoxide dismutase (SOD), glutathione-S-transferase (GST), glutathione peroxidase (GSH-Px), catalase, glutathione (GSH) E, flavonoid pigments, polyphenols and other physiologically active substances to protect against free radical damage. However, diabetic complications such as diabetic retinopathy, stroke, myocardial infarction, chronic renal failure, peripheral neuropathy, and hyperlipidemia occur due to the excessive production of free radicals due to hyperglycemia. Recently, researches have been conducted to prevent tissue damage through natural substances such as edible and medicinal plants for diabetes prevention and treatment and to have a defensive system in the body.

On the other hand, Moringa is a soybean plant, which is a tropical plant that edits the entire tree, including leaves, fruits, seeds, flowers, stems and roots. Moringa oleifera is the most widely known. Moringa oleifera is a very ideal source of protein, containing 40% of the moringa leaves protein, which is about twice as much as milk. In addition, the content of vitamins, minerals, and amino acids such as four times as much calcium as milk, four times as much vitamin A as carrots and three times as much as three times as the amount of bananas per gram of gum is higher than those of other plants. Especially, methionine It can be used as a source of cystine. Korean Patent Laid-Open No. 10-2013-0088224 (composition for external application for skin containing Moringa seed extract) as a prior art related to Moringa, etc., but the prior art discloses a composition for external application for skin, Radical scavenging ability, and blood glucose lowering ability.

Accordingly, the present inventors have searched for a new natural material without any side effects while having excellent diabetes prevention and therapeutic effect, and found that Moringa extract has antioxidant activity and radical scavenging activity and purifies metabolic syndrome caused by diabetes, The present invention has been completed.

It is an object of the present invention to provide a new use of moringa extract.

It is an object of the present invention to provide an antioxidative composition containing an extract of Moringa as an active ingredient, a composition for lowering blood glucose, a health functional food for preventing or improving diabetes and diabetic complications, And to provide a pharmaceutical composition for preventing or treating diabetic complications.

In order to achieve the above object, the present invention provides an antioxidative composition comprising a moringa extract as an active ingredient.

The moringa extract is characterized by being extracted from leaves, stems or seeds of Moringa.

The Moringa extract is characterized in that it is extracted from leaves, stems or seeds of Moringa using a solvent extraction method.

In addition, the present invention provides a composition for lowering blood glucose levels, comprising a moringa extract as an active ingredient.

The moringa extract is characterized by being extracted from leaves, stems or seeds of Moringa.

The Moringa extract is characterized in that it is extracted from leaves, stems or seeds of Moringa using a solvent extraction method.

Further, the present invention provides a health functional food for preventing or ameliorating diabetes and diabetic complications containing Moringa extract as an active ingredient.

The moringa extract is characterized by being extracted from leaves, stems or seeds of Moringa.

The Moringa extract is characterized in that it is extracted from leaves, stems or seeds of Moringa using a solvent extraction method.

The health functional food is characterized by being manufactured in the form of beverage, tea bag, capsule, powder or tablet.

The present invention also provides a pharmaceutical composition for preventing or treating diabetes mellitus and diabetic complications containing moringa extract as an active ingredient.

According to the present invention, it is possible to provide a moringa extract having excellent radical scavenging activity and antioxidation activity and having a hypoglycemic effect.

The present invention also provides an antioxidative composition comprising an extract of Moringa as an active ingredient, a composition for reducing blood glucose, a health functional food for preventing or ameliorating diabetes and diabetic complications, There is an effect of providing a pharmaceutical composition for preventing or treating diabetic complications. The health functional foods and pharmaceutical compositions for diabetes and diabetic complications according to the present invention have an effect of controlling the blood sugar level of the diseased person to a normal level and preventing diabetes in a normal person.

Figure 1 is an HPLC histogram of flavonoid content analysis of Moringa leaf extract.
Figure 2 is an HPLC histogram of flavonoid content analysis of Moringa stem extract.
Figure 3 is an HPLC histogram of flavonoid content analysis of Moringa seed extract.
Figure 4 is a graph of DPPH radical scavenging activity of moringa ethanol (top) and hydrothermal (bottom) extracts by site.
FIG. 5 is a graph showing the ORAC activity of the Moringa hot-water extract by site.
Figure 6 is a graph of hydroxy radical scavenging activity of moringa ethanol (top), hydrothermal (bottom) extract at each site.
FIG. 7 is a graph showing the activity of SOD-like active substances in the moringa ethanol (upper) and hot water (lower) extracts.
Figure 8 is a graph of H ₂ O ₂ radical scavenging activity of moringa ethanol (top), hydrothermal (bottom) extract by site.
FIG. 9 is a graph showing the change in glycated hemoglobin according to the Moringa wage.
FIG. 10 is a graph showing the blood glucose change according to the Moringa wage.

Hereinafter, the present invention will be described in detail.

The present invention provides an antioxidative composition containing a moringa extract as an active ingredient.

In addition, the present invention provides a composition for lowering blood glucose levels, comprising a moringa extract as an active ingredient.

Further, the present invention provides a health functional food for preventing or ameliorating diabetes and diabetic complications containing Moringa extract as an active ingredient.

The present invention also provides a pharmaceutical composition for preventing or treating diabetes mellitus and diabetic complications containing moringa extract as an active ingredient.

Diabetes is caused and promoted by the abnormal increase of free radicals, and antioxidants are involved in the signal transduction of specific genes to maintain normal blood sugar. In diabetic patients, the production of lipid peroxides in vivo is further accelerated as compared with that of normal subjects. This results in excessive production of free radicals (O 2 -, HO-), which leads to protein degradation, cell dysfunction such as chromosome aberration and erythrocyte destruction, And causes complications such as diabetic retinopathy, stroke, myocardial infarction, chronic renal failure, peripheral neuropathy and hyperlipemia. Therefore, the Moringa extract having antioxidative and hypoglycemic effects according to the present invention has an effect of preventing and treating diabetes mellitus as well as prevention and treatment of diabetes mellitus.

In addition, the Moringa extract according to the present invention has an excellent antioxidative activity, and can effectively prevent odor and flavor of food caused by oxidation, rancidity of the fattening product and discoloration of food. Therefore, by combining the above-mentioned Moringa extract with various conventional foods, it can be used to preserve these foods or to maintain the freshness and quality of food for a long period of time.

According to one embodiment of the present invention, the Moringa extract is extracted from Moringa leaves, stems or seeds.

The above-mentioned Moringa extract can be extracted from the leaves, stems or seeds of Moringa using a solvent extraction method, and water, a lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, dipropylene glycol, glycerin, At least one solvent selected from the group consisting of acetone, ethyl acetate, chloroform, methylene chloride, butyl acetate, diethyl ether, dichloromethane and hexane may be added.

Preferably, the leaves, stems or seeds of Moringa are dried and then added with a lower alcohol such as methanol, ethanol or butanol in a hot water or concentration of 50 to 80% at a temperature of 80 to 150 DEG C, which is 5 to 20 times the dry weight, The extracted effect shows the optimum effect. The moringa extract extracted by the above method contains various physiologically active substances and is excellent in antioxidant and antidiabetic effects and has a high absorption in the body and is useful for prevention and treatment of diabetes and diabetic complications by removing active oxygen and lowering blood sugar .

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 merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Examples. Preparation of moringa extract

Moringa leaves, stems and seeds were prepared and dried, respectively. Water corresponding to a weight of each leaf, stem, and seed was added at a weight corresponding to 10 times the weight and subjected to hot water extraction at 100 ° C for 2 hours. The extracts of each of the moringa leaves, stems, and seeds obtained by repeating the above extraction process three times were concentrated, filtered, dried and stored at 20 ° C.

In addition, Moringa leaves, stem and seed extracts were prepared as described above, and the solvent was extracted at 37 ° C using 70% ethanol.

Experimental Example 1. Analysis of physiologically active substances of Moringa extract

The flavonoids contained in each of the hot water and ethanol extracts of Moringa leaves, stems, and seeds prepared in the Examples were measured by HPLC (Shimadzu, Japan). As a result, in the case of Moringa leaf extract, (Fig. 1 to Fig. 3). ≪ tb > < TABLE >

HPLC was carried out on a Shimadzu SPD-10A (Kyoto, Japan) equipped with an LC-10AD pump, a DGU-10A deformation, a refractive index (RI) detector and a SIL-10AD autoinjector, and the separation was carried out on a ZORBAX Eclipse Plus C18 column 150 mm, 5 μm, Agilent Technologies, USA). Elution was carried out with a gradient solvent system using 1% glacial acetic acid (solvent A) and methanol (solvent B) as the mobile phase and the solvent was maintained at 80:20 (A / B) for 3 minutes and 65:35 (A / B) and maintained for 14 minutes, changed to 0: 100 (A / B) for 25 minutes, and maintained for 5 minutes. The flow rate was 1.0 mL / min at room temperature and monitored at 334 nm and the sample injection volume was 20 mL.

Experimental Example 2. Antioxidative Activity Analysis of Moringa Extract

(1) DPPH radical scavenging ability

100 μL of 0.15 mM DPPH solution (dissolved in 99% methanol, brown bottle, refrigerated) and 100 μL of sample were mixed and reacted at room temperature and dark place for 30 minutes. Absorbance was measured at 517 nm (blank: 100 μL of DPPH + methanol 100 [mu] L).

As a result, the moringa leaf extract showed the highest DPPH radical scavenging ability as shown in the table below, and the activity of the hydrothermal extract was higher than that of the ethanol extract (see FIG. 4).

(2) ORAC activity

The oxygen radical absorbent capacity (ORAC) is a measure of the rate of decrease of the fluorescent light caused by the formation and disappearance of peroxy radicals. The AAPH solution (300 mM) in 75 mM potassium phosphate buffer (pH 7.0) was stored at 4 ° C and the fluorescein sodium salt was dissolved in 50 nM phosphate buffer (0.1 mM EDTA). Fluorescein was allowed to stand in a 37 ° C water bath for 15 minutes, and the samples, fluorescein, and AAPH were added to the final volume ratio of 0.01, 2.7, and 0.02 ml, respectively, and the fluorescence was measured at 37 ° C every 5 minutes for 60 minutes using a fluorescence spectrophotometer. At this time, excitation wavelength was measured at 485 nm and emission wavelength was measured at 520 nm, and the result was expressed in μM TE (trolox equivalent).

As a result, the activity was higher than that of the reference substance Trolox at all sites, and the leaf hot water extract showed the highest activity at 1.0 mg / mL to 1323.161 μM TE (see FIG. 5).

(3) Hydroxy radical scavenging ability

0.1 mL of 10 mM FeSO ₄ / EDTA solution, 0.1 mL of 10 mM EDTA, 0.1 mL of 10 mM 2-deoxyribose, 0.1 mL of sample, 0.5 mL of 0.1 M phophate buffer (pH 7.4) and 0.1 mL of 10 mM H ₂ O ₂ After reacting for 4 hours at 37 ° C, 0.5 mL of 2.8% TCA solution was added, and the mixture was heated on a water bath at 100 ° C. for 10 minutes, and then quenched to measure the absorbance at 532 nm.

As a result, when the hot water and ethanol extracts of Moringa were added at different concentrations, the activity of hydroxy radical scavenging activity was increased as the concentration of the sample was increased as shown in the table below. In the ethanol extract, Showed superior hydroxy radical scavenging ability in leaves than seeds (see Fig. 6).

(4) Activity of SOD-like active substance

SOD (superoxide dismutase) is an enzyme that eliminates active oxygen, and it is known that it is important to determine how much SOD is adapted to the amount present. In addition, a low molecular substance which is not produced in the body but can suppress the reactivity of superoxide is referred to as SOD-like active substance. This substance is fundamentally different from the enzyme, SOD, because it does not convert steady state oxygen by floating change of superoxide. However, since it acts like SOD in that it protects the living body from reactive oxygen by suppressing the reactivity of superoxide, Like active substance. Since SOD is an enzyme, it is very expensive to produce by genetic engineering, and has a disadvantage that its effect is rapid and rapid, while its stability is rapidly reduced. On the other hand, SOD-like active substances are phytochemicals of low molecular weight and can be ingested through food, and when they are separated and extracted, they are relatively inexpensive and stable in the body and slow in effect It appears to be long. After adding 20 μL of sample, 100 μL of Tris-HCl buffer, and 20 μL of pyrogallol, the reaction was carried out at 25 ° C. for 10 minutes. Then, 100 μL of 1 N HCl was added and absorbance was measured at 420 nm.

As a result, the activity of Moringa leaf was the highest, among which the activity of hydrothermal extract was the highest at 347.931% (see FIG. 7).

(5) H ₂ O ₂ radical scavenging ability

0.1 mL of each sample, 0.1 mL of 0.1 M PBS, and 20 μL of 20 mM H ₂ O ₂ were sequentially added to the 96-well plate, followed by reaction at 37 ° C for 5 minutes. Then, 0.03 mL of 1.25 mM ABTS and 0.03 mL of 1 unit / mL peroxidase Was added and reacted at 37 ° C for 10 minutes and absorbance was measured at 405 nm.

As a result of addition of hot water and ethanol extracts of Moringa by concentration, the activity increased as the concentration of the sample increased as shown in the table below. In the ethanol extracts from the hot water extract, H ₂ O ₂ radical scavenging ability (See Fig. 8)

Experimental Example 3. Analysis of Antidiabetic Activity of Moringa Extract

(1) animal preparation

The type 2 diabetic animal model used in this experiment was a 8-week-old male C57BL / KsJ-db / db mouse and eight heterozygous C57BL / KsJ-db / + mice And then used in the experiment. This study was conducted in accordance with the Guidelines for the Management and Use of Laboratory Animals (NRC) after approval of the Ethical Committee of Animal Experiments at Konkuk University (Approval No .: KU13073). Animals were housed in a mouse cage (220 x 200 x 145 mm) for 2 animals. The animal room was maintained at a temperature of 23 ± 2 ° C, a relative humidity of 55 ± 2%, a light cycle of 12 hours (07:00 to 19:00 ) And the illumination was adjusted to 150 ~ 300 lux.

(2) glycosylated hemoglobin change

Glycated hemoglobin concentration HbA1C was measured by Ezi Aewon (Infopia, Korea) using reflectometry after sampling the tail vein after fasting the experimental animals for 12 hours on the first day of feeding. The mean HbA1C of the control group was 3.68% and the HbA1C of the experimental group was 8.51% on average. After 8 weeks, the control mice showed no significant changes in glycosylated hemoglobin before and after the moringa ingestion, whereas the experimental diabetic mice showed an increase of 0.41% from 8.57% to 8.92% in the mice not receiving moringa The mice consuming moringa decreased 7.2% from 8.45% to 7.84%, indicating that the intake of moringa decreased the level of glycated hemoglobin (see FIG. 9). The moringa extract fed to mice in this experiment is a hot-water extract of Moringa leaves.

(2) Changes in blood sugar

Fasting blood glucose was measured once every two weeks by fasting the experimental animals for 12 hours and then taking blood from the tail vein using a glucose meter (Infopia, Korea). On the first day of the experiment, the mean blood glucose level of the control group was 210.53 mg / dL and the blood glucose level of the experimental group was 452.38 mg / dL. On the other hand, after 8 weeks, the control mice showed no significant change in blood glucose levels before and after the moringa ingestion, whereas the diabetic mice were 472.33 mg / dL to 490.77 mg / dL in the mice not taking moringa 12.56%, while that of mice consuming moringa decreased by 14.45% from 449.83 mg / dL to 384.83 mg / dL, indicating that the intake of moringa decreased blood glucose levels (see FIG. 10). The moringa extract fed to mice in this experiment is a hot-water extract of Moringa leaves.

Experimental Example 4. Statistical Analysis

Statistical analysis was performed using PRISM (GraphPad Software, La Jolla, CA, USA). All experiments were repeated 3 times. The comparison of the measurements between the groups was done by one-way analysis of variance (ANOVA). Significance was given when the confidence interval was less than p <0.05.

Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (11)

An antioxidative composition comprising a moringa extract as an active ingredient.
The method according to claim 1,
Wherein said Moringa extract is extracted from leaves, stems or seeds of Moringa.
The method according to claim 1,
Wherein said Moringa extract is extracted from leaf, stem or seed of Moringa using a solvent extraction method.
A composition for lowering blood glucose levels, comprising a moringa extract as an active ingredient.
5. The method of claim 4,
Wherein the moringa extract is extracted from leaves, stems or seeds of Moringa.
5. The method of claim 4,
Wherein the moringa extract is extracted from leaf, stem or seed of Moringa using a solvent extraction method.
A functional food for preventing or ameliorating diabetes and diabetic complications containing moringa extract as an active ingredient.
8. The method of claim 7,
The above-mentioned Moringa extract is characterized by being extracted from leaves, stems or seeds of Moringa, and a health functional food for preventing or ameliorating diabetes and diabetic complications.
8. The method of claim 7,
Wherein the moringa extract is extracted from leaf, stem or seed of Moringa using a solvent extraction method, and a health functional food for preventing or improving diabetic and diabetic complications.
8. The method of claim 7,
Wherein said health functional food is manufactured in the form of beverage, tea bag, capsule, powder or tablet.
A pharmaceutical composition for preventing or treating diabetes and diabetic complications containing moringa extract as an active ingredient.

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