KR101216032B1 - Pharmaceutical composition for the treatment and prevention of disease relating diabetes - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating diabetes-related diseases. The present invention is a pharmaceutical composition for preventing and treating diabetes-related diseases, in the mineral concentrate containing a mineral selected from one or more selected from the group consisting of potassium, calcium, sodium and magnesium mistletoe, scabies, fern, birch, blue rice, barley, It includes raw materials selected from the group consisting of onions, beets, apples and persimmons. According to the present invention, through the pharmacological component extracted from Sanyacho by the minerals and minerals contained in the composition, various harmful toxins accumulated in the body can be removed to promote smooth metabolism of the human body. Therefore, it is possible not only to effectively prevent and treat diabetes, but also to prevent complications caused by diabetes.

Description

Pharmaceutical composition for the prevention and treatment of diabetes-related diseases {PHARMACEUTICAL COMPOSITION FOR THE TREATMENT AND PREVENTION OF DISEASE RELATING DIABETES}

The present invention relates to a composition for preventing and treating diabetes-related diseases, and more particularly, to a pharmaceutical composition for preventing and treating diabetes-related diseases using natural substances.

Diabetes is a disease in which excess glucose is excreted in the urine due to increased glucose content in the blood or abnormalities in carbohydrate metabolism, and metabolic disease causing abnormalities in protein and lipid metabolism and electrolyte metabolism. Diabetes causes hyperglycemia due to lowered secretion of insulin and defects in action that normalize blood glucose levels.

In general, diabetes is a chronic disease that indicates hyperglycemia due to impaired homeostatic control of glucose concentration in the blood, and is caused by type 1 diabetes and obesity, lack of exercise, smoking, and drinking habits caused by poor insulin secretion in the pancreas. It is classified as type 2 diabetes due to increased peripheral tissue resistance to insulin. In particular, the incidence of type 2 diabetes continues to increase with the increase in the obese population.

Most diabetic patients tend to coexist with insulin secretion disorder and insulin resistance. In this case, rapidly elevated blood glucose causes structural metabolic abnormalities due to the non-enzymatic glycosylation process. Therefore, oxidative stress in the body increases, and the activity of the antioxidant enzyme system in the body decreases, causing complications such as atherosclerosis and hypertension. In addition, diabetes is known to cause atherosclerotic vascular disease complications such as coronary artery disease, cerebrovascular disease, peripheral vascular disease, etc., and mortality and morbidity are increasing accordingly.

Therefore, in order to prevent and treat diabetes and complications, it is important not only to control blood glucose, but also to reduce lipid peroxide production and oxidative stress.

However, since a commonly used medicine is a chemical medicine made of a chemical artificial compound, it lacks reducibility in the living body and generates free radicals in the body, damaging DNA and causing resistance and various side effects. This is being triggered.

Therefore, there is a need for a therapeutic composition that obtains antidiabetic activity from natural plants having physiological activity and a method of treatment using the same.

The present invention has been proposed to meet the above demands, and an object of the present invention is to provide a pharmaceutical composition for the prevention and treatment of diabetes-related diseases comprising a raw solution obtained by mixing and fermenting wild vegetables with mineral concentrate as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing and treating diabetes-related diseases, including mistletoe, scabies, in a mineral concentrate containing at least one mineral selected from the group consisting of potassium, calcium, sodium and magnesium, It is characterized in that it comprises a stock solution fermented by mixing at least one raw material selected from the group consisting of fire, birch, blue rice, crushed barley, onion, beet, apple and persimmon as an active ingredient.

According to the present invention, it is possible to provide a pharmaceutical composition for the prevention and treatment of diabetes-related diseases comprising as an active ingredient a fermentation of fermented by mixing wild grass with mineral concentrate. In particular, through the pharmacological component extracted from Sanyacho by minerals and minerals contained in the composition, it is possible to promote various metabolism of the human body by removing various harmful toxins accumulated in the body. Therefore, it is possible not only to effectively prevent and treat diabetes, but also to prevent complications caused by diabetes.

1 is a flow chart illustrating a method for preparing a pharmaceutical composition for the prevention and treatment of diabetes-related diseases according to an embodiment of the present invention.

Hereinafter, the most preferred embodiments of the present invention will be described with reference to the accompanying drawings so that those skilled in the art may easily implement the technical idea of the present invention.

The pharmaceutical composition for treating diabetes mellitus according to an embodiment of the present invention is a stock solution fermented by mixing raw materials including mistletoe, scabies, brash, birch, blue rice, barley, onion, beet, apple and persimmon in seawater concentrate as active ingredients. Include.

Here, the seawater concentrate is a mineral concentrate including potassium (K), calcium (Ca), sodium (Na), and magnesium (Mg), with a total of 20 to 30% by weight of potassium, calcium, sodium, and magnesium based on the total weight of the mineral concentrate. It is preferred to be included. In addition, when the mineral concentrate contains all of potassium, calcium, sodium and magnesium, it is preferable that the potassium concentration is 45 to 50% by weight, 1 to 3% by weight of calcium, 45 to 50% by weight of sodium and the rest of magnesium. In particular, the weight ratio of potassium and sodium is 1: 1, and the weight ratio of calcium and magnesium is more preferably 2: 1.

Of course, the mineral concentrate also contains phosphorus (P), iron (Fe), silicon (Si), manganese (Mn), zinc (Zn), chromium (Cr), copper (Cu), selenium (Se), molybdenum (Mo) ), Bromine (Br), iodine (I), barium (Ba) and the like may be further included. The mineral concentrate having such a composition is preferably obtained by concentrating seawater, but the present invention is not limited thereto. In addition, a mineral concentrate including at least one mineral selected from the group consisting of potassium, calcium, sodium, and magnesium may be used in various ways. Once obtained, the therapeutic pharmaceutical compositions can be prepared.

Raw materials (sanyacho) that are mixed and fermented in mineral concentrates include substances that are rich in physiological activities such as various vitamins, enzymes, minerals, and fiber, and are used for blood purification, detoxification, diuresis, antibacterial, anti-inflammatory, tonic, antipyretic, analgesic, antioxidant, It is preferable that it is excellent in anticancer etc., and it is more preferable to include mistletoe, scabies, fern, birch, blue mussel, crushed barley, onion, beet, apple, and persimmon.

Mistletoe scientific name is' Viscum album var. It is a coloratum, and belongs to the mistletoe family of dicotyledonous plants. It lives on oak, teal, chestnut, and sesame. Contains oleanolic acid, saponin, amirin, biscuin, gum, etc. It is used for medicinal purposes. In oriental medicine, stems and leaves are used for molting, rapist, toothache, swelling, pain, low back pain, postpartum syndrome, frostbite, and arteriosclerosis. .

Ogapi refers to the bark of the roots, stems and branches of the Acanthopanax sessiliflorum Seeman or the same plant. Carbohydrates, minerals, iron, fat, vitamins, etc. are contained in the outpost, and have pharmacological effects such as immune enhancement, antioxidant, anti-fatigue, anti-high temperature, anti-irritant action, endocrine function control, blood pressure control, anti-radioactivity, detoxification effect.

Saree belongs to the legumes with the scientific name 'Lespedeza bicolor'. It grows in mountains and fields and is 2-3m high. Stems stand upright, branching is split a lot, leaves are displaced, and 3 small leaves come out. Chin leaves are thin and long, dark brown, about 5mm long. Small leaves are egg-shaped or upside down. The outer side is dark green with snowy hair on the back and flat edges. It contains alkaloids, flavonoids, ascorbic acid, tannins, and saponins.

Birch has the scientific name 'Betula platyphylla var. japonica '. The bark of birch contains about 35% of betulin, about 35% of various fatty acids, and about 7% of tannin. The endothelium contains about 11% of tannins. Leaves contain 0.25% of betafolientriol, triterpenoids, and 5.19% of tannins. In oriental medicine, birch bark is called baekhwapi (白樺 皮) is used for diuretic, analgesic, fever.

Cheongmirae is a vine shrub with Cheongmirae vine and its scientific name is 'Smilax china'. Flowers are reddish green and bloom in summer. Fruits are round, about 1cm in diameter, ripen in red from September to October, and are called Myeongmyeong or Mt. Fruits are edible and young sprouts are eaten as herbs. Roots are effective for diuresis, detoxification, gusts, etc. Used for arthritis, back pain, and boils. The blue future contains about 40% of various saponins composed of parrillin, smilacin, diosgenin, and the like.

Covered barley refers to barley that does not fall even after the grain inside and outside the grain (입자 外 穎) of maturation. Compared with rice, B6 contains about 5 times, pantothenic acid about 3 times, and folic acid about 14 times higher, and iron and minerals are also contained in large amounts.

Onion is a biennial herb of the monocotyledonous liliaceae with the scientific name 'Allium cepa'. 100g of onion contains about 90% water, about 8g of sugar and about 1.0g of protein. It contains minerals such as potassium, calcium, iron, phosphorus, sodium, dietary fiber, and folic acid, and it contains cyclic chlorine, gisalides (sulfur compounds), pectin, flavonoids, selenium, and quercetin. In particular, the scales contain minerals such as calcium and phosphoric acid together with various vitamins, and are effective in removing harmful substances in the blood.

Beet is scientific name 'Beta vulgaris' and it is a biennial plant of the subfamily of the dicotyledons. The main stem grows to about 1m and branches are branched. Leaves from the roots are thick, soft, oval or oval. The leaves from the stem are long oval or lanceolate with pointed ends. The leaves are light green to some reddish green and the surface is very shiny.

Apples are the fruit of the apple tree, a deciduous arborescent plant of the dicotyledonous plant Rosaceae. As an alkaline food, the main ingredient is carbohydrate, relatively low in protein and fat, and rich in minerals such as vitamin C, potassium, sodium and calcium. Potassium is effective in preventing and treating high blood pressure because it releases salt in the body, and it is rich in fiber that cleans the intestines and stimulates the secretion of gastric juice to help digestion and increase iron absorption. Relaxing sedation is good for insomnia, anemia, headaches are also effective.

Persimmon is the main component of the sugar is 15 to 16%. High in glucose and fructose, rich in vitamins A and B. 100g of persimmon contains 30 to 50mg of vitamin C, and also contains pectin and carotenoids.

According to the present invention, by mixing and fermenting the wild vegetable raw material in the mineral concentrate, it is possible to obtain a stock solution from which the pharmacological component of the raw material is extracted by the mineral. Therefore, it is possible to provide a pharmaceutical composition for treating diabetes comprising the stock solution as an active ingredient.

Hereinafter, a method for preparing a pharmaceutical composition for treating diabetes mellitus according to an embodiment of the present invention will be described with reference to the accompanying drawings.

1 is a flow chart illustrating a manufacturing process of a pharmaceutical composition for treating diabetes mellitus according to an embodiment of the present invention.

Referring to Figure 1, the present invention is to concentrate the seawater in order to obtain a mineral concentrate (S11), mixed with the raw material including mistletoe, scabies, bark, birch, blue rice, crushed barley, onion, beet, apple and persimmon in the mineral concentrate Step S12, fermenting the mineral concentrate mixed with the raw material to obtain a stock solution (S13) and preparing a pharmaceutical composition for treating diabetes containing the stock solution as an active ingredient (S14).

Concentration step (S11) comprises the step of concentrating the seawater to 5 to 6 times the original seawater volume through natural evaporation at 25 to 30 ℃ in evaporation to obtain a mineral rich mineral concentrate. Through this, it is preferable to obtain a mineral concentrate containing 20 to 30% by weight of potassium, calcium, sodium and magnesium, based on the total weight of the mineral concentrate.

In the mixing step S12, the raw materials are mixed with the mineral concentrate. Here, the raw grasses described above may be used or mixed, respectively. When used in combination, each component is preferably used at least 1% by weight or more, when all are used in combination, mistletoe 8 to 12% by weight, Ogapi 8 to 12% by weight, 8 to 12% by weight 8-12% by weight, birch 8-12%, barley 8-12%, onion 8-12%, beet 8-12%, apples 8-12% and the remaining persimmon is preferably used Do. In addition, red cabbage may also be used.

In addition, the mixing ratio of the mineral concentrate and the raw material is preferably 2: 1 to 3: 1 by weight, more preferably 70:30.

Fermentation step (S13) ferment the mineral concentrate mixed with the raw material at 10 to 25 ℃ for 1 to 3 years. At this time, while the mineral concentrate mixed with the raw material is aged in the fermentation process, the pharmacological component of the raw material is extracted by the mineral contained in the mineral concentrate. After the fermentation step is completed, the stock solution containing the minerals and pharmacological components of wild grasses is completed.

In the step S14 of preparing a pharmaceutical composition for treating diabetes, a pharmaceutical composition for treating diabetes containing a stock solution as an active ingredient is prepared.

On the other hand, the mineral concentrate, the mixed solution, the stock solution and the like obtained in each step may be selectively sterilized, and may be filtered.

In the pharmaceutical composition for preventing and treating diabetes-related diseases according to the present invention, the stock solution is in any amount as long as it can prevent and treat diabetes within the range that it is not toxic to the human body depending on the type or progression of diabetes. May be included. Preferably it can be used in the range of 0.1 to 10% by weight, more preferably from 1 to 3% by weight based on the total weight of the composition.

In addition, the composition of the present invention can be used in combination with any compound that is effective in the prevention and treatment of diabetes, in addition to the wild vinegar extract.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier in addition to the stock solution as an active ingredient, such a pharmaceutically acceptable carrier is commonly used in pharmaceutical preparations, lactose, dextrose, sucrose , Sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxy Benzoate, talc, magnesium stearate, mineral oil, and the like.

The pharmaceutical compositions of the present invention may also further comprise lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like known in the art as additives. The additive may be included in about 0.1% to about 20% by weight based on the total weight of the composition.

On the other hand, the pharmaceutical composition of the present invention may be administered orally or parenterally, and may be formulated in unit dose form, as formulated in a general manner in the art.

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate and sucrose in the stock solution of the present invention. (Sucrose) or lactose (Lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, 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 sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the non-aqueous solvent and the suspension solvent, 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.

In addition, the pharmaceutical composition of the present invention may be mixed with distilled water for injection to have a salt concentration similar to that of a body fluid and may be prepared as a Ringer's solution.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier or incorporated into a multi-dose container. In this case, the formulation may be in the form of a solution, a suspension or an emulsion, or may include elexirs, extracts, powders, granules, tablets, ointments, and the like.

The pharmaceutical composition of the present invention has a daily dosage in the range of 1 to 3 g / kg body weight, and can be administered once or in several divided doses. However, since the dosage of the pharmaceutical composition of the present invention is determined in accordance with various related factors such as the route of administration, the age, sex, weight of the patient, the severity of the patient, the duration of use, and the like, the dosage may be determined in any aspect of the present invention. It should not be understood to limit the scope.

Hereinafter, a method for preparing a pharmaceutical composition for treating diabetes according to an embodiment of the present invention will be described in detail with reference to the embodiment. In addition, with reference to the test example to look for the effect of administering a pharmaceutical composition for treating diabetes according to an embodiment of the present invention to a mammal. In particular, in the test example, the effect of adding 1%, 3% and 5% of the stock solution to the feed for mammals will be described in detail.

Example

1,000 L of seawater was evaporated at 30 ° C. in an evaporation basin to obtain a mineral concentrate concentrated to 200 L.

Mistletoe, scabies, bracken, birch, blue rice, barley, onions, beets, apples and persimmons were all washed thoroughly and then dried. Next, 10% by weight of mistletoe, 10% by weight of bark, 10% by weight of birch, 10% by weight of birch, 10% by weight of barley, 10% by weight of barley, 10% by weight of onion, 10% by weight of beet, 10% by weight of apple and persimmon 10 Weighing by weight to prepare a raw material.

Subsequently, the mineral concentrate and the raw material were placed in a large container, and then capped and fermented at room temperature for 1 year. At this time, the weight of the mineral concentrate and the weight of the raw material was to be 70:30 ratio.

Subsequently, the liquid in which the fermentation was completed was filtered to obtain a stock solution.

Test Example

The stock solution prepared by the above-described embodiment was sealed and stored at −4 ° C., and added to the prepared feed for breeding of rats.

1. Breeding of experimental animals Diet

Sprague-Dawley male rats with an average weight of 150 ± 10 g were distributed from Samtaco (Osan, Korea).

The temperature of the feeding room was 22 ℃, the humidity was 50 ± 5% and the contrast period was adjusted to 12 hours (07: 00 ~ 19: 00). During the breeding period, water and feed were freely fed. The experimental animals were adapted to solid feed for the first week after the pre-incubation, and after 2 weeks of preliminary breeding with a normal diet containing 10% corn oil, the animals were divided into groups according to their weight. 9 animals were divided into 5 groups so that the body weight was similar to the experimental breeding for 4 weeks. The dietary composition of each experimental group during the experimental breeding period is shown in Table 1 below.

Experimental breeding was divided into normal group (Normal) and STZ (streptozotocin) which did not induce diabetes. In addition, the diabetic group was fed the diabetic control group (Diabetic), which fed the feed without the undiluted solution, the 1% feed group (Diabetic-1%) who fed the feed containing 1% of the stock solution, and the feed added with 3% the stock solution. Were compared to 3% fed group (Diabetic-3%) and 5% fed group (Diabetic-5%).

(%, diet) group
ingredient Normal
(normal) Diabetes control
(Diabetic) 1%
(Diabetic-1%) 3%
(Diabetic-3%) 5%
(Diabetic-5%) carbohydrate
(Starch) 39.74 39.74 39.74 39.74 39.74 casein
(Casein) 20 20 20 20 20 Cellulose
(Cellulose) 5 5 5 5 5 Sucrose
(Sucrose) 10 10 10 10 10 Vitamin Mix ^{1 )} One One One One One Mineral mix ^{2 )} 3.5 3.5 3.5 3.5 3.5 DL-mesionine
(DL-methionine) 03. 0.3 0.3 0.3 0.3 Choline Hydrogen Tartrate (Choline bitartrate) 0.2 0.2 0.2 0.2 0.2 Corn oil
Corn oil 7 7 7 7 7 Undiluted solution - - One 3 5

Vitamin mixture ^{1 )} : AIN-76 ^TM

Mineral mixture ^{2 )} : AIN-76 ^TM

2. Diabetes induction in experimental animals

To induce diabetes, STZ, which is known to act specifically on the β-cells of the pancreas and does not affect other organs, was used. STZ is not known to cause absolute insulin deficiency in pancreatic β-cells, but is known to damage the insulin secretory response system to glucose in the early stages, leading to hyperglycemia.

STZ was dissolved in 0.01 M citrate buffer (pH 4.5) and injected once (50 mg / kg body weight) intraperitoneally to induce diabetes. The normal group was intraperitoneally injected with the same amount of citrate buffer solution.

To determine the cause of diabetes, 48 hours after STZ injection, blood was collected from the tail vein on an empty stomach, and blood glucose was measured using a blood glucose meter (Active Blood Glucose Meter, ACCU-CHEK ^?, USA). When the experiment started breeding.

3. Dietary intake , Dietary efficiency  And weight measurement

 During the experiment, the diet was fed at 5 pm every day, and the amount of food intake was calculated by checking the remaining amount at 10 am the next day. Water was supplied freshly every day with tap water. Body weight was measured once a week at a specific time. The total weight gain (g) during the four weeks of experimental breeding was divided by the number of experimental days to calculate the average weight per day, and the food efficiency ratio (FER) equals the weight gain. Calculated by dividing the dietary intake over the period.

4. Treatment of experimental animals

After 16 hours of fasting on the last day of experimental breeding, anesthetized with ether, blood was collected from the heart, and blood was obtained. Blood was coagulated in ice water for about 30 minutes and centrifuged at 3,000 rpm for 15 minutes (Mega 17R, HANIL, Korea) to separate serum. Each tissue (soy, heart, kidney, spleen, testes) is extracted immediately after blood collection and washed thoroughly with cooled saline solution, then drained with absorbent paper, weighed, stored at -70 ℃ and used for analysis. It was.

5. Blood analysis

 (1) blood sugar measurement

Glucose was measured using glucose reagent (AM 201-k, Asan, Korea) for measuring glucose from serum obtained on the final day of experimental breeding. After adding 3 mL of the enzyme reagent to 20 μL of serum, the cells were incubated at 37 ° C. for 5 minutes, and the absorbance was measured at a wavelength of 500 nm using the serum-free group as a control. Blood glucose content was expressed in mg / dL by standard calibration curve.

(2) Determination of glycated hemoglobin content

Glycosylated hemoglobin content was measured by hemoglobin A1c (Glycosylated hemoglobin, Asan, Korea). EDTA treated whole blood was measured by microcolumn chromatography using a Helena laboratories kit reagent. The glycated hemoglobin content was calculated by the absorbance ratio of glycated hemoglobin to total hemoglobin.

(3) lipid content analysis

The total lipid content of serum was incubated for 15 minutes at 37 ° C. by adding phospho-vanillin reagent to 20 μL of serum according to the method of Frings et al. Absorbance was measured at. Total cholesterol content was measured with a kit reagent for measuring total cholesterol (AM 202-k, Asan, Korea). Triglyceride content was measured using a kit reagent for measuring triglycerides (AM 157S-k, Asan, Korea). Phospholipids were measured using a reagent for measuring phospholipids (SICDIA L PL, Eiken, Japan).

HDL-C (High density lipoprotein cholesterol) content was measured using a kit reagent for measuring HDL-C (AM 203-k, Asan, Korea).

Low density lipoprotein cholesterol (LDL-C) content was calculated by the formula of total cholesterol-(HDL-C + triglyceride / 5).

Very low density lipoprotein cholesterol (VLDL-C) content was calculated by the formula of total cholesterol-(HDL-C + LDL-C).

Atherosclerotic index (AI) was calculated by the formula of total cholesterol-(HDL-C) / (HDL-C) by the method of Haglund (Haglund).

Cardiac risk factor (CRF) was calculated by the formula of total cholesterol / HDL-C by Kang et al.

(4) GOT and GPT Activity Assay

GOT (glutamic oxaloacetic transaminase) and GPT (glutamic pyruvic transaminase) activities of serum were measured using a blood analyzer (DRI-Chem 3500I) with a kit for measuring GOT (P 3150, Fugi, Japan) and a kit for measuring GPT (P 3250, Fugi, Japan). , Fugi, Japan), and expressed as karmen units per mL of serum.

(5) Determination of thibarbituric acid reactive substances (TBARS) content

To measure the lipid peroxide content in serum, 1 / 12N sulfuric acid solution and 10% PTA (phosphotungstic acid) were sequentially added to 100 μL of serum, followed by centrifugation at 4,000 rpm for 10 minutes. After centrifugation, 1 mL of distilled water and TBA reagent were added to the precipitate and reacted for 60 minutes in a 95 ° C. water bath. The resulting lipid peroxide was transferred to butanol, and the absorbance was measured at 532 nm.

6. Component analysis of liver tissue

Hepatic tissue lipids were prepared by adding a chloroform: methanol mixture (C: M = 2: 1, v / v) to 0.5 g of hepatic tissue according to Floch et al. Tissue grinder (DAIHAN WOS01010). , Korea) and extracted a certain amount of lipids to measure the total lipids, total cholesterol and triglycerides with each kit kit for measurement.

(1) Determination of lipid peroxide content in liver tissue

Uchiyama and Mihara's method and Lee's method were analyzed. In other words, 1.5 g KCl solution was added to 1 g of liver tissue and ground with a homogenizer to make a 10% homogenate. Then, 0.5 mL was taken, and 3 mL of 1% phosphoric acid and 0.6 mL of 1 mL were added. TBA was added and mixed well. The mixture was heated for 45 minutes in a 95 ° C. water bath, 4 mL of butanol was added to extract the coloring material, and the absorbance (OD _{535 -520} ) of the butanol layer was centrifuged at 3,000 rpm for 10 minutes. TBARS content of hepatic tissue was calculated from a standard calibration curve using tetraethoxypropane (1,1,3,3-tetraethoxypropane) as a standard solution.

(2) Determination of glycogen content in liver tissue

The glycogen content of the liver tissue was quenched after adding 20% KOH solution to a predetermined amount of tissue and heating it in a water bath at 100 ° C. for 20 minutes. After mixing with 95% ethanol, the mixture was heated for another 5 minutes, centrifuged at 3000 rpm for 4 minutes, and the supernatant was removed. Distilled water was added to the precipitate, mixed with 0.2% anthrone solution, heated, cooled, and the absorbance was measured at 620 nm. Glycogen content was calculated from the standard calibration curve obtained by the same method using glucose.

The experimental results are expressed as mean and standard deviation, and statistical significance was compared by Duncan's multiple range test at p <0.05 after one-way analysis of variance using SPSS package. Analyzes were made.

7. Results

(One) Dietary intake  And weight change

The weight, dietary intake and dietary efficiency of diabetic rats according to the feeding of the composition for treating diabetes according to an embodiment of the present invention are shown in Table 2 below.

Final weight (g) Dietary Intake
(g / day) Total weight gain
(g / 4 weeks) Dietary Efficiency (FER ^* ) Normal (Normal) 331.67 ± 17.04 ^d 20.62 ± 0.33 ^NS 136.50 ± 13.75 ^d 23.64 ± 2.48 ^d Diabetes control
(Diabetic) 278.50 ± 15.36 ^c 20.83 ± 0.37 71.33 ± 16.48 ^bc 12.26 ± 2.93 ^bc 1%
(Diabetic-1%) 271.67 ± 38.78 ^bc 20.57 ± 0.76 77.67 ± 34.16 ^c 13.54 ± 5.98 ^c 3%
(Diabetic-3%) 245.50 ± 12.93 ^ab 20.63 ± 0.51 47.33 ± 15.88 ^ab 8.18 ± 2.67 ^ab 5%
(Diabetic-5%) 224.33 ± 19.02 ^a 20.87 ± 0.09 29.83 ± 12.83 ^a 5.10 ± 2.18 ^a

^* FER (Food Efficiency Ratio) = total weight gain / dietary intake

STZ-induced diabetic laboratory animals are characterized by reduced cellular glucose utilization, metabolic characteristics of starvation, and recovery of body weight. In particular, since liver glycogen content is reduced by about 4.5 times in STZ-induced diabetes, weight loss during diabetes is also used as an indicator of the degree of diabetes.

As a result, the final weight was decreased in the feed group compared to the diabetic control group, but the difference was less in the 1% feed group. The total dietary intake for 4 weeks was not different in all groups.

In particular, the total weight gain during the four weeks of experimental breeding was 71.33 g in the diabetic control group, compared with 136.50 g in the normal group, and 29.83 to 77.67 g in the undiluted group, which was higher in the 1% group than in the 5% group. In addition, the dietary efficiency (FER) was 12.26 in the diabetic control group, which was 13.54 in the 1% group, and the difference was small in the 3% and 5% group.

(2) long-term weight

The results of measuring the weight of the organs extracted after 4 weeks of experimental breeding are shown in Table 3 below. The weight of the organs is expressed based on the weight of the liver, heart, kidneys, spleen and testes based on the weight of 100 g.

(tissue g / 100g body weight) Liver Heart Kidney Spleen Testis Normal (Normal) 4.55 ± 0.45 ^a 0.32 ± 0.02 ^a 0.71 ± 0.03 ^a 0.19 ± 0.01 ^NS 1.00 ± 0.05 ^a Diabetes control
(Diabetic) 6.20 ± 0.60 ^bc 0.37 ± 0.04 ^ab 0.93 ± 0.13 ^b 0.22 ± 0.03 1.23 ± 0.10 ^b 1%
(Diabetic-1%) 5.56 ± 0.50 ^b 0.35 ± 0.06 ^ab 0.98 ± 0.18 ^b 0.21 + 0.04 1.25 ± 0.06 ^b 3%
(Diabetic-3%) 6.61 ± 1.10 ^c 0.37 ± 0.06 ^ab 1.080.11 ^b 0.23 ± 0.04 1.31 ± 0.18 ^b 5%
(Diabetic-5%) 6.13 ± 0.42 ^bc 0.38 ± 0.05 ^b 1.27 ± 0.15 ^c 0.19 + 0.03 1.35 ± 0.13 ^b

^Ac values with the same subscript in the column are not significant at p <0.05

NS: No significant meaning

As a result, the weight of the liver was the largest in the diabetic control group and small in the 1% feeding group, but the difference was small. The weights of heart, kidney and testis were not significantly different from those of the diabetic control group, 1% and 3%. Zira had no difference in weight in all experimental groups.

Through this, the weight of the organs increased when diabetes occurred compared to the normal group, it was determined that the feed of the stock solution according to an embodiment of the present invention does not significantly affect the weight change of the organs in the body when diabetes occurs. That is, even if the stock solution is 1%, 3%, 5%, it was confirmed that there is no problem in the long-term weight at all.

(3) blood sugar content

The blood glucose content was measured from blood collected after 4 weeks of experimental breeding and is shown in Table 4 below.

Blood Glucose (mg / dL) Normal (Normal) 149.75 ± 18.38 ^a Diabetes control
(Diabetic) 224.70 ± 22.23 ^c 1%
(Diabetic-1%) 164.60 ± 16.51 ^ab 3%
(Diabetic-3%) 168.76 ± 27.49 ^ab 5%
(Diabetic-5%) 187.18 ± 18.12 ^b

^Ac values with the same subscript in the column are not significant at p <0.05

STZ damages the β-cells of the pancreas, reducing insulin secretion, and abnormal metabolism of sugar metabolism reduces sugar utilization, resulting in hyperglycemia by the action of glucose synthase. The hypoglycemic effect of natural plants has been reported to increase the secretion of insulin by further stimulating pancreatic β-cells or to regenerate pancreatic cells due to the presence of insulin-like substances in plants.

As a result, as a result of feeding the stock solution according to an embodiment of the present invention to diabetic rats induced by STZ, blood sugar was significantly reduced in the 1%, 3%, and 5% feeding groups compared to the diabetic control group. In particular, the 1% feeding group showed almost the normal rats, and the 1% and 3% feeding effect was found to be the best.

This effect may be due to hypoglycemia in 1% and 3% feeding groups, which stimulates β-cells to increase insulin secretion, or to consume and consume intravascular glucose that cannot move into cells. There is little transient difference.

Therefore, it is expected that blood glucose levels in diabetic rats can be normalized if the stock solution is continuously fed 1% or 3%.

(4) Glycation  Hemoglobin content

After 4 weeks of experimental breeding, the results of measuring the glycated hemoglobin content in blood are shown in Table 5 below.

Glycated Hemoglobin (mg / dL) Normal (Normal) 2.57 ± 0.58 ^a Diabetic Control 4.79 ± 0.71 ^c 1% feeding group (Diabetic-1%) 3.06 ± 0.48 ^ab 31% Feed group (Diabetic-3%) 3.09 ± 0.22 ^ab 51% Feed group (Diabetic-5%) 3.59 ± 0.76 ^b

^Ac values with the same subscript in the column are not significant at p <0.05

Diabetes degrades hemoglobin content by lowering kidney function, and high levels of blood sugar combine with hemoglobin to form glycated hemoglobin. Therefore, the measurement of the content of glycated hemoglobin is used as one of the diabetic diagnostic method, it means that the reduction of the glycated hemoglobin content in diabetes is effective in controlling diabetes.

When the stock solution according to an embodiment of the present invention was fed to diabetic rats, the glycated hemoglobin content was significantly reduced in the feed group (1, 3, 5%) compared to the diabetic control group. In particular, the 1% and 3% feeding groups were measured at almost the same level as normal mice. However, unlike the 1% and 3% feed groups, the content of glycated hemoglobin was slightly increased in the 5% feed group. Therefore, the feed amount of the undiluted solution may be appropriate when diabetes is induced.

(5) blood lipid content

After 4 weeks of experimental breeding, the results of analyzing lipid components in blood are shown in Table 6 below.

(mg / dL) Total lipid Total cholesterol Triglyceride Phospholipids Normal (Normal) 205.78 ± 7.73 ^a 60.29 ± 1.67 ^a 50.57 ± 4.08 ^a 210.29 ± 25.58 ^a Diabetes control
(Diabetic) 321.85 ± 9.66 ^c 66.35 ± 5.90 ^bc 59.85 ± 4.50 ^b 264.00 ± 11.15 ^b 1%
(Diabetic-1%) 238.10 ± 21.74 ^b 62.60 ± 3.04 ^ab 57.65 ± 3.59 ^b 211.43 ± 31.72a 3%
(Diabetic-3%) 250.85 ± 11.66 ^b 68.80 ± 4.19 ^cd 60.24 ± 3.83 ^b 247.27 ± 25.62 ^b 5%
(Diabetic-5%) 309.95 ± 10.70 ^c 72.55 ± 2.96 ^d 62.23 ± 4.47 ^b 243.22 ± 8.90 ^ab

^Ac values with the same subscript in the column are not significant at p <0.05

Lipid metabolism abnormalities observed in diabetic patients include elevated triglycerides, cholesterol and decreased levels of HDL-cholesterol. In the diabetic group, the total cholesterol content in the blood is increased because carbohydrates are not used as energy sources but because free fatty acids are used as energy to synthesize cholesterol. It is reported to increase.

As a result, total lipids were higher in the diabetic group than in the normal group, but the undiluted feeding group was significantly reduced compared to the diabetic control group. In particular, the 1% feed group had the lowest total lipid content.

The total cholesterol content was not significantly different from the diabetic control group and the undiluted feeding group, but it was the lowest in the 1% feeding group among the 1, 3, and 5% feeding groups.

Triglyceride was not different between the diabetic control group and the undiluted feeding group, and the effect of reducing triglyceride content in blood was small.

Phospholipids content in the 1% feed group was similar to that of the normal group, and the phospholipid content is estimated to be restored to normal level by the feed.

Therefore, in diabetic rats, it was observed that the total lipid, cholesterol and phospholipid content in the blood decreased after the feeding of undiluted solution, which would be effective in reducing the incidence of complications such as hyperlipidemia and arteriosclerosis caused by diabetes. It is feed.

(6) HDL - Cholesterol  content, LDL Cholesterol content, AI  And CRF  Indices

After 4 weeks of experimental breeding, the results of analyzing blood HDL-cholesterol content, LDL-cholesterol content, AI and CRF index are shown in Table 7 below.

HDL-C
(mg / dL) LDL-C
(mg / dL) AI CRF Normal (Normal) 35.96 ± 1.30 ^c 14.21 ± 0.77 ^a 0.68 ± 0.02 ^a 1.68 ± 0.02 ^a Diabetes control
(Diabetic) 31.90 ± 1.85 ^a 23.29 ± 7.41 ^bc 1.14 ± 0.27 ^cd 2.14 ± 0.27 ^cd 1%
(Diabetic-1%) 34.96 ± 2.12 ^bc 16.11 ± 2.19ab 0.79 ± 0.06 ^ab 1.79 ± 0.06 ^ab 3%
(Diabetic-3%) 34.67 ± 2.64 ^bc 22.08 ± 5.15 ^bc 1.00 ± 0.19 ^bc 2.00 ± 0.19 ^bc 5%
(Diabetic-5%) 31.73 ± 2.81 ^ab 28.37 ± 5.91 ^c 1.30 ± 0.26 ^d 2.30 ± 0.26 ^d

^Ad values with the same subscript in the column are not significant at p <0.05

It has been reported that the reduction of HDL-cholesterol in the blood is a phenomenon derived from lipid metabolism abnormalities occurring in diabetes. Serum HDL-cholesterol is an indicator of anti-arteriosclerosis. Unlike lipoproteins, serum HDL-cholesterol separates LDL-cholesterol from the blood vessel walls and metabolizes it in the liver for energy or promotes excretion of the body. It is known to reduce the risk. LDL-cholesterol is a causative agent of diabetic complications. Insulin deficiency decreases LDL receptors, which prevents the efficient removal of LDL in the body, resulting in increased LDL-cholesterol levels in the blood. In particular, adult diseases from the circulatory system are caused by atherosclerosis, and the atherosclerosis index (AI) is used as an early indicator of atherosclerosis. The atherosclerosis index and cardiovascular risk index are mainly associated with risk of atherosclerosis and cardiovascular disease. It is widely used as a risk index, and increasing levels of AI and CRF mean a higher incidence of atherosclerosis and cardiovascular disease.

CRF represents the ratio of total cholesterol content to HDL-cholesterol. The excessively high total cholesterol content compared to the HDL-cholesterol content means that the lipoprotein content of LDL-cholesterol and VLDL-cholesterol other than HDL-cholesterol is high. This means that the probability of developing cardiovascular disease increases.

Experimental results showed that HDL-C content in diabetic rats was significantly increased, which was similar to that of normal group in 1% and 3% fed group. The LDL-cholesterol (LDL-C) content was decreased in the feed group compared to the diabetic control group, but was effectively decreased in the 1% and 3% groups.

Atherosclerosis index (AI) was highest in diabetic control group and decreased in 1% and 3% feeding group. In particular, in the 1% feeding group, the arteriosclerosis index was lowered to the level of the normal group, and it was estimated that the incidence of arteriosclerosis, which is one of the diabetic complications, could be reduced by the feeding of the stock solution.

The risk of developing cardiovascular disease index (CRF) was 1% and 3% lower than those of the diabetic control group. Therefore, less than 3% of the stock solution may reduce the risk of complications in diabetic rats.

(7) Protein, albumin and globulin content in the blood

After 4 weeks of experimental breeding, blood protein, albumin and globulin content measurement results are shown in Table 8 below.

(mg / dL) Total protein albumin globulin Normal (Normal) 6.41 ± 0.39 ^NS 4.45 ± 0.28 ^NS 1.95 ± 0.19 ^NS Diabetes control
(Diabetic) 6.21 ± 0.16 4.35 ± 0.08 1.87 ± 0.22 1%
(Diabetic-1%) 6.51 ± 0.39 4.50 ± 0.15 2.02 ± 0.43 3%
(Diabetic-3%) 6.29 ± 0.12 1.35 ± 0.59 1.94 ± 0.64 5%
(Diabetic-5%) 6.09 ± 0.12 4.26 ± 0.20 1.84 ± 0.30

NS: No significant meaning

The decrease in total protein in diabetic rats is due to poor utilization of glucose in the blood, which is used as a calorie source, and this has been reported to result in weight loss. If diabetes is not treated, the breakdown of proteins is increased, and the consumption of these proteins becomes clearer as the diabetes lasts longer.

The experimental results showed that there was no significant difference in the total protein, albumin and globulin contents according to the feeding of the undiluted solution during 4 weeks of diabetes. This was confirmed. In particular, the total protein, albumin and globulin content in the 1% feeding group was measured to be higher than the normal group, if the continuous intake of the stock solution is believed to be able to compensate for the consumption of body protein during diabetes.

(8) lipid peroxide content in the blood

After 4 weeks of experimental breeding, blood lipid peroxide content measurement results are shown in Table 9 below.

TBARS (mmol / dL) Normal (Normal) 41.44 ± 4.58 ^a Diabetic Control 60.33 ± 5.94 ^b 1% feeding group (Diabetic-1%) 57.19 ± 3.37 ^b 3% Feeding Group (Diabetic-3%) 67.19 ± 14.69 ^b 5% feeding group (Diabetic-5%) 90.89 ± 10.06 ^c

Diabetes induction is known to increase the content of triglycerides in the body produces a modified LDL less binding to the LDL receptor and is easily oxidized by free radicals in the blood to produce more lipid peroxides.

The results showed that the lipid peroxide content in blood was significantly higher in the diabetic control group than in the normal group, and the lipid peroxide content of the 1% and 3% feeding groups was decreased, but there was no significant difference. In the 5% feeding group, the lipid peroxide content was increased more than the diabetic control group.

(9) In soy sauce organization GOT , GPT  activation

After 4 weeks of experimental breeding, the results of GOT and GPT activity analysis in hepatic tissues are shown in Table 10 below.

(u / dL) GOT GPT Normal (Normal) 53.50 ± 3.32 ^ab 2.75 ± 0.96 ^a Diabetes control
(Diabetic) 62.25 ± 7.72 ^c 4.50 ± 1.00 ^b 1%
(Diabetic-1%) 52.75 ± 5.91 ^a 5.00 ± 1.15 ^b 3%
(Diabetic-3%) 57.50 ± 2.52 ^abc 6.75 ± 0.50 ^c 5%
(Diabetic-5%) 61.25 ± 3.77 ^bc 8.75 ± 1.50d

^Ad values with the same subscript in the column are not significant at p <0.05

GOT and GPT are present in large amounts in hepatocytes and are widely used enzymes for the diagnosis of liver disease. If the enzyme is released into the blood as the necrosis of liver cells and the destruction of liver tissue proceed due to hepatotoxicity in the serum, the activity of the enzyme increases in the plasma. Therefore, the activity of this enzyme is used as an indicator of liver damage.

GOT and GPT activities were significantly higher in the diabetic control group than in the normal group, but GOT was significantly decreased in the 1% feeding group compared to the diabetic control group, so the 1% feed solution was effective against the liver damage caused by diabetes. I think there will be.

(10) lipid content in liver tissue

After 4 weeks of experimental breeding, the results of measuring lipid content in liver tissue are shown in Table 11 below.

(mg / g wet liver) Total lipid Total cholesterol Triglyceride Normal (Normal) 10.41 ± 0.93 ^a 1.77 ± 0.26 ^NS 8.03 ± 0.56 ^b Diabetes control
(Diabetic) 12.13 ± 0.57 ^b 1.87 ± 0.17 10.22 ± 0.82 ^c 1%
(Diabetic-1%) 11.07 ± 0.59 ^ab 1.44 ± 0.33 6.36 ± 0.97 ^a 3%
(Diabetic-3%) 11.40 ± 1.45 ^ab 1.53 ± 0.07 4.90 ± 1.67 ^a 5%
(Diabetic-5%) 11.58 ± 0.31 ^ab 1.75 ± 0.36 5.05 ± 1.07 ^a

^Ac values with the same subscript in the column are not significant at p <0.05

NS: No significant meaning

In STZ-induced diabetic rats, lipid components accumulate in the liver tissue and blood fat outflow is increased. In the case of diabetes, the conversion of blood fat to triglyceride has been reported to increase the content of triglyceride in the liver tissue.

As a result, total lipids were decreased in the feed group compared to the diabetic control group, but there was little difference. In particular, the 1% feed group had the lowest total lipid content. The total cholesterol content was also lower in the untreated group than in the diabetic control group, but there was no significant difference. Triglycerides were significantly lower in the undiluted group than the diabetic control group, indicating that the triglyceride content in the hepatic tissue was decreased by the undiluted feed. In addition, the 1% feeding group has lower total lipid and cholesterol content than the 3% and 5% feeding groups.

(11) lipid content in kidney tissue

After 4 weeks of experimental breeding, the results of measuring lipid content in kidney tissue are shown in Table 12 below.

(mg / g wet liver) Total lipid Total cholesterol Triglyceride Normal (Normal) 7.95 ± 1.08 ^NS 3.61 ± 0.16 ^a 5.50 ± 1.22a Diabetes control
(Diabetic) 8.98 ± 1.94 4.32 ± 0.12 ^b 11.95 ± 1.86 ^b 1%
(Diabetic-1%) 7.34 ± 0.75 3.30 ± 0.80 ^a 7.38 ± 2.78 ^a 3%
(Diabetic-3%) 7.42 ± 0.80 3.24 ± 0.48 ^a 7.10 ± 0.94 ^a 5%
(Diabetic-5%) 7.47 ± 1.36 3.08 ± 0.38 ^a 6.81 ± 1.83 ^a

^Ab values with the same subscript in the column are not significant at p <0.05

NS: No significant meaning

In the case of diabetes, high blood sugar damages the capillaries of the kidneys, which lowers kidney function, which causes proteinuria to be excreted in the urine protein. Fat accumulation in tissues is also caused by the same mechanism as the liver.

As a result, total lipids were decreased in the feed group than in the diabetic control group, but there was no significant difference. Total cholesterol content was significantly decreased in the feed group compared to the diabetic control group. However, there was no difference according to the feed amount of the stock solution.

Triglycerides also showed the same trend as total cholesterol. Compared with the lipid content in the liver (see Table 11), the lipid content in the kidneys was lower in the total lipid and triglyceride content, which is similar to the report that the kidneys are less sensitive than the liver in lipid accumulation and lipid peroxide production in tissues. It was.

(12) Lipid peroxide content in liver tissue

After 4 weeks of experimental breeding, the results of measuring the lipid peroxide content in the liver tissue are shown in Table 13 below.

TBARS (mmol / g wet liver) Normal (Normal) 139.56 ± 11.09 ^b Diabetes control
(Diabetic) 155.85 ± 9.29 ^c 1%
(Diabetic-1%) 137.71 ± 12.53 ^b 3%
(Diabetic-3%) 106.59 ± 2.23 ^a 5%
(Diabetic-5%) 105.48 ± 10.61 ^a

^Ac values with the same subscript in the column are not significant at p <0.05

Lipid peroxidation, which causes cancer, arteriosclerosis and aging, is considered to be a kind of cell damage mechanism. In addition, it is recognized as the most important mechanism that indicates the physiological physiological phenomenon or the degree of tissue damage caused by various toxic compounds, drugs, or diseases, resulting from increased oxidative stress of cells in tissues and reduction of antioxidant defense ability in vivo. Because it becomes. In general, the increase of oxidative stress in diabetes increases the lipid peroxide in the tissue, and there is a report that increased the lipid peroxide MDA (malondialdehyde) content in the kidney and hepatic tissue as well as the vascular system.

As a result of measuring the lipid peroxide content that causes the oxidative damage, aging and other degenerative diseases of the tissue, the lipid peroxide content in the hepatic tissue was higher than that of the diabetic control group. In addition, the content of lipid peroxide was reduced to a level similar to that of the normal group in the 1% feeding group, it was measured to have a lower content in the 3% and 5% feeding group. Therefore, it is estimated that the feeding of the stock solution during diabetes can protect the tissue by inhibiting the oxidation of lipids accumulated in the liver tissue.

(13) glycogen content in liver tissue

After 4 weeks of experimental breeding, the results of measuring glycogen content in liver tissue are shown in Table 14 below.

Glycogen content Normal (Normal) 19.85 ± 4.40 ^c Diabetes control
(Diabetic) 12.46 ± 0.99 ^a 1%
(Diabetic-1%) 14.96 ± 2.90 ^b 3%
(Diabetic-3%) 14.00 ± 1.37 ^b 1%
(Diabetic-5%) 15.05 ± 1.42 ^b

^Ac values with the same subscript in the column are not significant at p <0.05

When diabetes is induced by STZ, the glycogen synthase is inactivated due to the lack of insulin due to the destruction of β-cells of the pancreas, and the glycogen degrading enzyme is activated to decrease glycogen content in the body. In the case of liver, insulin resistance develops within 3 days after the administration of STZ, and the glycogen in the liver is reduced due to a significant effect on diabetes compared to muscle.

As a result, the glycogen content of the liver during diabetes was lower than that of the normal group, but the glycogen content was significantly higher than that of the diabetic control group. Therefore, the feed of diabetic solution is believed to be effective in repairing the body tissue by increasing the glycogen content in the liver.

Through the above test results, it can be seen that there is an excellent effect on diabetes in the feed group less than 2% of the stock solution. In particular, the stock solution according to the present invention is effective in improving lipid metabolism, such as cholesterol as well as diabetes, and it can be seen that the lipid metabolism effect is indirectly effective in diabetes.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the embodiments are for the purpose of illustration only and are not to be construed as limitations. In addition, those skilled in the art will recognize that various embodiments are possible within the scope of the technical idea of the present invention.

Claims (6)

A seawater concentrate containing potassium (K), calcium (Ca), sodium (Na), and magnesium (Mg) is mixed with raw materials including mistletoe, scabies, barks, birch, blue rice, bran, onion, beet, apple, and persimmon Fermented stock solution contains as an active ingredient
Pharmaceutical composition for preventing and treating diabetes.
The method of claim 1,
The stock solution contains 1 to 3% by weight based on the total weight of the composition
Pharmaceutical composition for preventing and treating diabetes.
The method of claim 1,
The stock solution is,
Concentrating the seawater to obtain the seawater concentrate;
Mixing raw materials including mistletoe, scallop, brash, birch, bluish green, crushed barley, onion, beet, apple and persimmon to the seawater concentrate; And
Fermenting the seawater concentrate mixed with the raw materials
Obtained by
Pharmaceutical composition for preventing and treating diabetes.
The method of claim 1,
The seawater concentrate,
Obtained by concentrating seawater 5 to 6 times as much as the original seawater through natural evaporation at 25 to 30
Pharmaceutical composition for preventing and treating diabetes.
The method of claim 1,
The seawater concentrate and the raw material is mixed in a ratio of 2: 1 to 3: 1 by weight
Pharmaceutical composition for preventing and treating diabetes.
The method of claim 1,
The stock solution is,
Obtained by fermenting the seawater concentrate mixed with the raw material at 10-25 ℃ for 1 to 3 years
Pharmaceutical composition for preventing and treating diabetes.

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