KR20210047780A - Pharmaceutical composition comprising the extract of prune as an effective component for prevention or treatment of diabetes and health functional food comprising the same - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and a health functional food for preventing or treating diabetes, containing an extract of Prunus domestica as an active ingredient, and more specifically, to a pharmaceutical composition and a health functional food for preventing or treating/alleviating diabetes through a strong β-amylase and α-glucosidase inhibitory activity, containing a hot water extract of 100 to 160 day-old Prunus domestica fruits after flowering as an active ingredient. The extract of Prunus domestica of the present invention as the pharmaceutical composition and the health functional food for preventing or treating/alleviating diabetes has excellent effects of being used as pharmaceuticals and health functional foods for preventing or alleviating diabetes through a strong β-amylase and α-glucosidase inhibitory activity as demonstrated through the examples herein. In addition, the Prunus domestica extract of the present invention has excellent thermal stability and no loss of starch degrading enzyme inhibitory activity even under acidic conditions of pH 2 and in plasma, and thus can be easily processed into various forms such as liquid, cream, powder, pill, tablet, etc., and can be very advantageously used in the pharmaceutical industry and the food industry.

Description

Pharmaceutical composition and health functional food for preventing or treating diabetes containing prune extract as an active ingredient {PHARMACEUTICAL COMPOSITION COMPRISING THE EXTRACT OF PRUNE AS AN EFFECTIVE COMPONENT FOR PREVENTION OR TREATMENT OF DIABETES AND HEALTH FUNCTIONAL FOOD COMPRISING THE SAME}

The present invention relates to a pharmaceutical composition and health functional food for the prevention or treatment of diabetes (diabetes) containing an extract of Prune (prune, Prunus domestica) as an active ingredient, and more specifically, Prune 100 to 160 days after flowering A pharmaceutical composition and health function for preventing or treating/improving diabetes through potent beta-amylase and alpha-glucosidase inhibitory activity containing hot water extract of fruit as an active ingredient It's about food.

Diabetes, one of the most common diseases in modern people, is a type of metabolic disease such as insufficient secretion of insulin or inability to function normally. It is characterized by high blood glucose concentration and causes various symptoms and signs due to high blood sugar. .

Diabetes is classified into type 1 and type 2. Type 1 diabetes is caused by the inability to produce insulin at all due to genetic effects, and type 2 diabetes is caused by poor insulin function and insulin resistance. ) Is known as the cause. In particular, type 2 diabetes is known to be largely affected by environmental factors such as high calorie, high fat, high protein diet, lack of exercise, and stress due to westernization of diet.

For the treatment of diabetes, insulin injection is essential in the case of type 1 diabetes, and lifestyle correction and drug treatment are necessary in the case of type 2 diabetes. Nide) and a delaying agent for carbohydrate absorption in the small intestine (glucobi: ingredient name-acarbose, bason: ingredient name-voglibose), and the like are used.

On the other hand, Prunus domestica is a fruit of a shrub belonging to the Rosaceae family, and is also called Western plum. It has excellent taste and aroma, and its origin is known as the Caucasus Mountains near the Caspian Sea in Western Asia. Prunes are externally distinguished from Asian plums (Prunus salicina , Prunus triflora ), which are called Japanese or Chinese plums, in shape and size, and are also clearly differentiated from rosacea plums (Prunus mume ) and apricots ( Prunus armeniaca). Prunes are mainly harvested before maturity, dried, processed into dried plums, and sold. This is because, unlike plums, the fruit surface is often cracked during maturity, which makes it difficult to harvest high-quality prunes. In addition, prunes do not rot easily, unlike plums, even when dried in the presence of seeds after harvesting. Potassium sorbate is made on the surface during the drying process, making it appear as though white powder was sprinkled on the surface, showing antibacterial activity, so it easily rots. There is a characteristic that does not work.

Prune contains a large amount of protein, lipids, vitamins, and minerals. Especially, it has a significant amount of dietary fiber, and has been used as a constipation improvement agent since the past. It is effective in improving eye health, osteoporosis, heart disease prevention, and hypertension. It is known from.

Studies related to prunes are insignificant compared to plums, and research on oriental plums ( Prunus salicina ) has been intensively conducted. In various research literature, Plum (Oriental Plum) has been reported to have antioxidant, antibacterial, and anti-inflammatory activities (Ye-Na Heo, 2007, Daegu University Master's Thesis), and nutritional and physicochemical components (Sung Yoon-Jeong, 2002, J. Kor. Soc. Agric. Chem. Biotechnol. 45, 134-137). In the case of furun, it has been reported that it induces growth inhibition and death of human colon cancer cells (Fujii Takashi et al., 2006, Nutr. Sci. Vitamonol. 52, 389-391), and antibacterial (Yaqeen Zahra, 2013, Pakistan Journal of pharmaceutical sciences 26: 409-414) and cancer cell killing effect (Miljic, Uros, 2016, Journal of the Institute of Brewing 122: 342-349), antioxidant activity (Usenik Valentina et al., 2013, J. Science of Food and Agriculture 93: 681-692) is known. In addition, potent antioxidant activity has also been reported in the leaves of furun (Stierlin Emilie et al., 2018, J. Sci. Food Agricul. 98, 726-736), and the extract oil from furun leaves harvested in August has potent antioxidant and butyrylcholinesterase inhibitory activity. It has been reported (Marco Bonsi, 2019, Antioxidant, 8, 2; doi:10.3390/antiox8010002), and it has been suggested that the prevention of neurodegenerative diseases caused by inflammation and death of neurons may be helpful. In addition, there have been reports of Purunusides with α-Glucosidase inhibitory activity from shoots of furun (Shaheen Kosar et al., 2009, Purunusides AC, α-Glucosidase Inhibitory Homoisoflavone Glucosides from Prunus domestica. Arch Pharm Res 32: 1705-1710), which is After drying the sprouts of the furun, it was extracted with ethanol, the extract was sequentially fractionated with an organic solvent, and then the butanol fraction was obtained through silica-gel chromatography to obtain a prunuside material, which is different from the antidiabetic activity in the furun fruit. Until now, the antidiabetic effect of pruned fruit has not been reported.

Patents related to prunes include "Prune juice production using cellulase" in Korean Patent No. 10-1731232, "Prune juice production using cellulase" in US-4371552, Japanese patent JP- 0238079 Tea manufacturing (Leaf tea and beverage therefrom and product from its extract evaporation to dryness), JP-0002967 tyrosinase activity inhibitor and cosmetic containing the same (Tyrosinase activity inhibitor and cosmetic containing the same) and JP-0002964 The use of cosmetics through hyaluronidase activity inhibitor and humectant and cosmetic containing the same, and the manufacturing method and use of furun juice as a urease inhibitor are registered in JP-0193477.

In addition, Korean Patent Application Publication No. 10-2007-0065665 "whitening cosmetic composition", No. 10-2003-0009739 "a pharmaceutical composition for the treatment of constipation", No. 10-2016-0089117 "Prune extraction and filtration method" , No. 10-2016-0089118 discloses "a feed additive for fisheries using residues extracted from prunes", and No. 10-2003-0096615 discloses "a composition of a functional food with excellent constipation improvement effect and a method for manufacturing the same". US Patent Publication No. US-0360062 discloses "prune-based nutrient-rich materials and related processes", and Japanese Patent Application Publication No. JP-0008938 discloses "antitumor agent containing extract of leaves of plants belonging to rosaceae prunus". However, to date, no studies on the antidiabetic activity of immature prunes have been known.

KR 10-2003-0009739 A KR 10-2016-0089118 A

Shaheen Kosar et al., 2009, Purunusides A-C, α-Glucosidase Inhibitory Homoisoflavone Glucosides from Prunus domestica. Arch Pharm Res 32: 1705-1710. Jiwoon Nam et al., 2018, Effects of Plums from Gimcheon on Streptozotocin Diabetic Rats. Journal of the Korean Society of Food Culture 33: 291-298.

The present invention was conceived to solve the problems of the prior art as described above, and the problem to be solved in the present invention is a pharmaceutical composition for preventing or treating/improving diabetes containing a furun extract as an active ingredient, and a health functional food. I want to provide.

In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention or treatment of diabetes containing a prune extract of the President variety as an active ingredient.

It is preferable that the President variety's prune is a fruit of 100 to 160 days after flowering.

In addition, the present invention provides a health functional food for the prevention or improvement of diabetes containing the prune extract of the President variety as an active ingredient.

It is preferable that the President variety's prune is a fruit of 100 to 160 days after flowering.

Prune extract as an active ingredient of the pharmaceutical composition for preventing or treating/improving diabetes and health functional food of the present invention, as demonstrated through the examples of the present specification, has strong beta-amylase and alpha-glucosidase inhibitory activity It has excellent effects that can be used as medicines and health functional foods for preventing or improving diabetes through. In addition, the purun extract of the present invention has excellent thermal stability and does not show any loss of starch degrading enzyme inhibitory activity even in the acidic condition of pH 2 and plasma, so it can be easily processed into various forms such as liquid, cream, powder, pills, tablets, etc. It can be very useful in the pharmaceutical industry and food industry.

1 is a view from the left showing the Prune maturation of the President, Big Purple, and Black King varieties used in the present invention and the maturing plums as a control,
Figure 2 shows the President variety prunes harvested in July (100 days after flowering), August (120 days after flowering) and September (160 days after flowering) from the left,
3 is a cross-sectional view of the President variety prunes harvested in July (100 days after flowering), August (120 days after flowering) and September (160 days after flowering) from the left.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention prepared extracts of various varieties (President, Big Purple, Black King) as an extract through a certain method in order to test the anti-diabetic efficacy of the extracts, and evaluated the anti-diabetic activity of the extracts. The antidiabetic activity of the President's Prune extract was confirmed, and the President's Prune extract was prepared for each harvesting period and the antidiabetic activity was evaluated. President Prune at 100 to 160 days after flowering, preferably 120 days after flowering, was very excellent. It was confirmed that it showed activity. The furun extract exhibits anti-diabetic activity by confirming that it does not exhibit any hemolytic activity against human red blood cells, but has excellent thermal stability and acid stability. It was intended to be used as a functional food.

Specifically, the present inventors in order to develop a pharmaceutical composition and health functional food for the prevention or treatment / improvement of diabetes using a furun containing antioxidant components such as polyphenol or anthocyanin in a large amount compared to ordinary fruits, President, As a result of preparing hot water extracts of Prunes of Big Purple and Black King varieties, respectively, and evaluating the antidiabetic activity of these extracts against beta-amylase and alpha-glucosidase. , Strong beta-amylase (β-amylase) and alpha-glucosidase (α-glucosidase) inhibitory activity was confirmed in President Prune extract 100 to 160 days after flowering, preferably 120 days after flowering, and the extract It was confirmed that the hemolytic activity on human red blood cells did not appear, so that it can be practically used.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating diabetes containing a prune extract of the President variety as an active ingredient.

Prunes of the President variety are preferably fruits 100 to 160 days after flowering, more preferably 120 days after flowering.

It is preferable that the Prune extract of the President variety is hot water or ethanol extract.

In addition, the present invention provides a health functional food for the prevention or improvement of diabetes containing the prune extract of the President variety as an active ingredient.

Prunes of the President variety are preferably fruits 100 to 160 days after flowering, more preferably 120 days after flowering.

It is preferable that the Prune extract of the President variety is hot water or ethanol extract.

Hereinafter, a method for preparing a furun extract of the present invention and an experiment of efficacy will be described in more detail.

The present invention comprises the steps of preparing an extract from furun; Evaluating the antidiabetic activity of the furun extract; And investigating the stability of the active extract.

"Prune extract" included in the composition of the present invention comprises the steps of removing seeds from the prune fruit; Extracting the prune fruit with an organic solvent; Filtering the extract using a filter net of 0.06 mm or less; And it can be obtained by the step of concentrating under reduced pressure.

The organic solvent used in the present invention is water (cold water, hot water), alcohol, anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, alcohol, propanol, ethyl acetate, etc.), and a mixed solvent of the lower alcohol and water. And the like may be used, and hot water or 95% ethanol extraction is most preferred.

The purun extract of the present invention may be prepared into a powder through a conventional powdering process such as drying under reduced pressure, freeze drying, or spray drying. They are not decomposed by various degrading enzymes in plasma, and they remain active even at a heat treatment of 100°C and a pH of 2 in the stomach.

The furun extract of the present invention exhibits potent beta-amylase and alpha-glucosidase inhibitory activity against starch degrading enzymes, and thus, type 1 diabetes, type 2 diabetes, or diabetic retinopathy, It can be used as a material for a pharmaceutical composition and health functional food related to the prevention and treatment/improvement of diabetic complications such as diabetic nephropathy.

The pharmaceutical composition containing the active ingredient of the present invention is formulated according to a conventional method for each purpose of use, oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and injections of sterile injectable solutions. It may be formulated and used in various forms such as, and may be administered orally or administered through various routes including intravenous, intraperitoneal, subcutaneous, rectal, and topical administration.

Such pharmaceutical compositions may further include a carrier, excipient, or diluent, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, Starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, amorphous cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil And the like. In addition, the pharmaceutical composition of the present invention may further include a filler, an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, a preservative, and the like.

In a preferred embodiment, 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, and the like, in the pharmaceutical composition. It is formulated by mixing sucrose, lactose, gelatin, and the like. Further, in addition to simple excipients, lubricants such as magnesium stearate and talc may be used.

As a preferred embodiment, as a liquid formulation for oral use, a suspension, a liquid solution, an emulsion, a syrup, etc. may be exemplified, and various excipients other than water and liquid paraffin, which are commonly used simple diluents, such as wetting agents, sweetening agents, Fragrances, preservatives, and the like may be included.

As a preferred embodiment, the preparation for parenteral administration may include a sterile aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized agent, a suppository, and the like. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyloleate. Injectables may contain conventional additives such as solubilizing agents, isotonic agents, suspending agents, emulsifying agents, stabilizing agents, and preservatives.

The active ingredient of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, drug activity, Sensitivity to the drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or administered in combination with another therapeutic agent, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

As a preferred embodiment, the effective amount of the active ingredient in the pharmaceutical composition of the present invention may vary depending on the age, sex, and body weight of the patient, and is generally 1 to 5,000 mg, preferably 100 to 3,000 mg per kilogram of body weight daily. Alternatively, it may be administered every other day or divided into 1 to 3 times a day. However, since it may increase or decrease according to the route of administration, the severity of the disease, sex, weight, age, etc., the dosage amount does not limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration can be expected and can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intracerebroventricular injection.

In the present invention, "administration" means providing a predetermined substance to a patient by any suitable method, and the route of administration of the pharmaceutical composition of the present invention is oral or parenteral through all general routes as long as it can reach the target tissue. It can be administered orally. In addition, the composition of the present invention may be administered using any device capable of delivering an active ingredient to target cells.

In the present invention, the "subject" is not particularly limited, but includes, for example, a human, a monkey, a cow, a horse, a sheep, a pig, a chicken, a turkey, a quail, a cat, a dog, a mouse, a mouse, a rabbit, or a guinea pig. And, preferably, it means a mammal, more preferably a human.

In addition, the health functional food of the present invention can be used in various ways such as foods and beverages effective in preventing or improving diabetes. Foods containing the active ingredient of the present invention include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. .

In general, the active ingredient of the present invention may be added in an amount of 0.01 to 15% by weight of the total food weight, and the health beverage composition may be added in an amount of 0.02 to 10g, preferably 0.3 to 1g, based on 100ml.

The health functional food of the present invention may contain the compound as an essential component in the indicated ratio as an additional component, as well as food supplementary additives acceptable for food, such as natural carbohydrates and various flavoring agents.

Examples of the natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and conventional sugars such as polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.

As the flavoring agent, natural flavoring agents such as stevia such as taumatin, rebaudioside A, or glysirhizin, and synthetic flavoring agents such as saccharin and aspartame may be used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention includes a variety of nutrients, vitamins, minerals, synthetic flavors and natural flavors, and other flavoring agents, coloring agents and thickeners, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloids. It may contain thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the health functional food of the present invention may contain pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is generally selected in the range of 0.01 to about 20 parts by weight per 100 parts by weight of the active ingredient of the present invention.

Hereinafter, the present invention will be described in more detail through examples. The following examples are only a preferred specific example of the present invention, and the scope of the present invention is not limited to the scope of the following examples.

[Example]

Example 1: Characteristics of Prunes Varieties and Harvested Mature Fruits

As for the prunes used in this example, mature President, Big Purple, and Black King Prunes cultivated at the Green Field Prune Farm in Waryong-myeon, Andong-si, Gyeongsangbuk-do, Korea in 2019 were used.In particular, in the case of the President, in addition to the maturation, 100 days after flowering, 120 Fruits of days and 160 days were additionally used. FIG. 1 is a photographic view of used prunes, and FIG. 2 is a photograph of fruits harvested at 100, 120, and 160 days after flowering, and FIG. 3 is a photographic view of the President prunes 100, 120 and 160 after flowering. It is a cross-sectional view of the first harvested fruit. The characteristics and physicochemical analysis results of each harvested Prune variety are shown in Tables 1 and 2.

The physicochemical analysis and color difference were measured by taking the juice after grinding the mature fruit. At this time, the color difference analysis was measured using a Hunter Color Difference meter (Super color SP-80 Colormeter, Tokyo Denshoku Co., Japan), and brightness. (White 100~0 black), redness (red 100~-80 green), and yellowness (yellow 70~-80 black) were measured. The chromaticity of the standard white board was set as 92.44 for L value, -0.06 for a value, and 1.35 for b value, and the average value was calculated by measuring three times per sample, and the color difference (△E) was calculated using the following equation. I did.

[Table 1] Weight and Size of Prunes Used by Variety

[Table 2] Analysis of Physicochemical Characteristics and Color Difference of Prunes Used by Varieties

First, as shown in Table 1, there was a difference in weight and diameter depending on the breed from maturity, and it was in the order of Big Purple <President <Black King. The pH measurement result of each Prune variety was 3.4, but the acidity of President and Black King was 0.23~0.24, while that of Big Purple was 0.43. In addition, brix also showed about twice as high as other blue varieties in Big Purple, and Big Purple was sweet and sour, showing the best sensory properties. On the other hand, as a result of chromaticity analysis, the brightness was similar, but the president showed high redness, and the big bubble showed high yellowness, and the overall color difference was similar to 76.2~77.3.

Example 2: Analysis of Hot Water Extraction Efficiency and Components of Prune Extract

Distilled water 10 times the weight of the fruits of various varieties harvested in Example 1 was added, heated at 100° C. for 1 hour, and filtered to prepare a purun extract. The prepared purun extract was concentrated under reduced pressure at 60°C to prepare a powder. Each extraction efficiency and component analysis results of the extract are shown in Table 3.

The content of total polyphenols, total flavonoids, total sugars and reducing sugars was measured by component analysis. For the total polyphenol content, 50 μl of Folin-ciocalteau and 100 μl of Na ₂ CO ₃ saturated solution were added to 400 μl of the extraction sample, and then allowed to stand at room temperature for 1 hour, and the absorbance was measured at 725 nm. Tannic acid was used as a standard reagent. For the total flavonoid content, each sample was extracted with methanol for 18 hours, 4 ml of 90% diethylene glycol was added to 400 μl of the filtered extraction sample solution, 40 μl of 1N NaOH was added again, and the absorbance was measured at 420 nm after 1 hour reaction at 37°C. . As a standard reagent, rutin was used. Reducing sugar was quantified by DNS method and total sugar was quantified by phenol-sulfuric acid method.

[Table 3] Analysis of Extraction Efficiency and Useful Components of Hot Water Extract of Prune Fruit of Various Varieties

As shown in Table 3, the hot water extraction efficiency was in the order of Big Purple> President> Blacking, and the average extraction efficiency was 12.6%. The total polyphenol content of the furun extract was high at 14.8 mg/g in Big Purple and 14.0 mg/g in Blacking, whereas it was relatively low in President at 10.7 mg/g. On the other hand, the total flavonoid content was in the order of blacking> president> big purple, and the total sugar and reducing sugar content was in the order of big purple> blacking> president.

Example 3: Evaluation of antidiabetic activity of furun extracts for various varieties

The antidiabetic activity of the furun extracts obtained in Example 2 was evaluated for beta-amylase inhibitory activity and alpha-glucosidase inhibitory activity, and the results are shown in Table 4.

First, β-amylase (4-alpha-D-glucan maltohydrolase) inhibitory activity was obtained by mixing 2.5 μl of a sample and 25 μl of β-amylase (0.25 U/ml) diluted with 50 mM phosphate buffer (pH 6.8) at 37°C for 10 minutes. After the first reaction, 25 μl of 0.5% soluble starch (Samchun Chemicals Co., Korea) was added and the second reaction was performed at 37°C for 10 minutes, and then the reaction was stopped by heating at 100°C for 5 minutes, and 150 μl of DNS in the reaction solution (3,5-dinitrosalicylic acid, Sigma Co., st. Louis, USA) solution was added, heated at 100° C. for 5 minutes to develop color, and then allowed to cool at room temperature. The color developing solution was calculated by measuring the absorbance at 540 nm.

Inhibition rate (%) = [1-(Enzyme activity after addition of sample/Enzyme activity after addition of control)] x 100

Meanwhile, α-glucosidase inhibitory activity was evaluated using pNPG (p-nitrophenol glucoside; Sigma Co., USA), and α-glucosidase (0.25 U/ml) diluted with 2.5 μl of sample and 50 mM sodium acetate buffer (pH 5.6). ) 25 μl of the mixture was first reacted at 37° C. for 10 minutes, and 25 μl of 1 mM pNPG solution was added, followed by the second reaction at 60° C. for 10 minutes. Thereafter, 25 μl of 1M NaOH was added to stop the reaction, and the inhibition rate was calculated by measuring the absorbance at 405 nm.

Inhibition rate (%) = [1-(Enzyme activity after addition of sample/Enzyme activity after addition of control)] x 100

[Table 4] Antidiabetic Activities of Prune Extracts by Various Varieties

As a result, as shown in Table 4, acarbose, an antidiabetic drug used in clinical practice, inhibited β-amylase and α-glucosidase by 62.7% and 73.9%, respectively, at a concentration of 0.5mg/ml, confirming the basis for clinical use. Prune extracts of various cultivars showed weaker β-amylase and α-glucosidase inhibitory activities than acarbose at 0.5 mg/ml, but President extract showed significant antidiabetic activity.

Example 4: Preparation of extracts and analysis of components of President Prune fruit by harvest time

The change of antidiabetic activity at each harvest time was evaluated for the President, which showed the best antidiabetic activity among mature and purun cultivars. The President was harvested in July (100 days after flowering), August (120 days after flowering), and September (160 days after flowering), and the size and physicochemical properties of fruit were carried out in the same manner as in Examples 1 and 2. And the content of useful ingredients were evaluated, and the results are shown in Tables 5, 6, and 7.

[Table 5] Weight and diameter of President Prune fruit by harvest time

[Table 6] Physicochemical Characteristics and Color Difference Analysis of President Prunes by Harvest Time

As shown in Table 6, the overall pH was 3.4, and the acidity was continuously decreased from 0.77 on the 100th day of flowering, and the acidity was 0.23 on the 160th day of flowering. In the case of brightness, there was little difference, but an increase in redness and a decrease in yellowness were observed at maturity.

[Table 7] Formulation and composition analysis of President Prune extract by harvest time

Meanwhile, as shown in Table 7, as the fruit matured, the hot water extraction efficiency increased, and the total sugar content increased. Specifically, the total polyphenol and total flavonoid content did not change significantly between 100 and 120 days after flowering, and at maturity, the total polyphenol and total flavonoid content decreased to a level of 60% compared to the unripe fruit. Therefore, useful physiological activity was expected to be excellent in unripe fruit 120 days after flowering.

Example 5: Evaluation of Antidiabetic Activity of President Prune Fruits by Harvest Time

The antidiabetic activity of the President's Prune fruit extract obtained in Example 4 by harvest time was evaluated in the same manner as in Example 3.

[Table 8] Antidiabetic Activity of President Prune Fruits by Harvest Time

As a result, as shown in Table 8, excellent β-amylase and α-glucosidase inhibitory activity was exhibited in President Prune fruits 100, 120 and 160 days after flowering, and in particular, very excellent β-amylase and It showed α-glucosidase inhibitory activity. Therefore, it was confirmed that the immature fruit and fruit of President Prune could be developed as an antidiabetic agent.

Example 6: Human red blood cell hemolytic activity of furun extract

Prune is a non-hazardous food that has been widely edible for a long time and has been secured. In the present invention, human red blood cell hemolytic activity was evaluated in order to evaluate the acute toxicity of President Purun extract at various harvesting periods, and the results are shown in Table 9.

At this time, the hemolytic activity was evaluated according to the previous report (Miseon Kim et al., 2014. J. Life Sci. 24: 515-521), and simply 100 μl of human red blood cells washed three times with PBS was added to a 96-well microplate and various 100μl of the sample solution of concentration was added and reacted for 30 minutes at 37°C. After that, the reaction solution was centrifuged for 10 minutes (1,500rpm), and 100μl of the supernatant was transferred to a new microtiter plate, and the degree of hemoglobin leakage due to hemolysis was measured at 414nm. I did. DMSO (2%) was used as a solvent control for the sample, and Triton X-100 (1 mg/ml) was used as an experimental control for hemolysis of red blood cells. The hemolytic activity was calculated using the following formula.

[Table 9] Human Red Blood Cell Hemolytic Activity of President Prune Extract by Harvest Time

As a result, as shown in Table 9, first, it was confirmed that DMSO used as a control did not show red blood cell hemolytic activity, and triton X-100 hemolytic 100% red blood cells at a concentration of 1 mg/ml. In addition, it was confirmed that amphotericin B, which is used as an anticancer agent and antifungal agent, hemolyzes more than 50% of red blood cells at a concentration of 0.025mg/ml.

On the other hand, President Prune extract by harvest time had no hemolytic activity at all. In addition, it was confirmed that the hemolytic activity did not appear at all even in the case of big bubble and blacking. Therefore, it is judged that the extract of the President's immature fruit will not exhibit a separate problem of inducing acute toxicity.

Example 9: Plasma, acid and thermal stability evaluation of furun extract

Plasma stability, thermal stability, and acid stability for antidiabetic activity were confirmed for the President's Purun extract 120 days after flowering obtained in Example 4 above. The samples were heat treated at 100° C. for 1 hour, treated at pH 2 (0.01M HCl) for 1 hour, and treated in plasma for 1 hour. I got it. Therefore, the furun extract is expected to maintain excellent antidiabetic activity during the digestion and absorption process and during the food manufacturing process.

Claims (4)

A pharmaceutical composition for the prevention or treatment of diabetes containing a prune extract of the President variety as an active ingredient. The pharmaceutical composition according to claim 1, wherein the President variety's prune is a fruit 100 to 160 days after flowering. Health functional food for preventing or improving diabetes containing the President variety's prune extract as an active ingredient. The health functional food according to claim 3, wherein the President variety's prune is a fruit 100 to 160 days after flowering.

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