A plant component hypoglycemic composition and its preparation method and application

Abstract

The present invention provides a plant component hypoglycemic composition and its preparation method and application, belonging to the field of biomedical technology. The pharmaceutical composition is composed of the following raw materials in parts by weight: 2-5 parts of mulberry polypeptide, 5-10 parts of polygonatum and 5-10 parts of eucommia male flower, and its preparation method is to first extract mulberry polypeptide from mulberry, then crush and decoct polygonatum and eucommia male flower, and then add Lactobacillus acidophilus and Bacillus subtilis in sequence for batch fermentation, and take the fermentation supernatant and freeze-dry it and mix it with mulberry polypeptide to obtain the hypoglycemic pharmaceutical composition of the present invention. The pharmaceutical composition prepared by the present invention has a significant inhibitory effect on α-glucosidase in vitro, has a good hypoglycemic effect in vivo, has a high utilization rate of medicinal materials and has no toxic side effects, and has broad market prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a plant component hypoglycemic composition, a preparation method and application thereof. Background Art

[0002] Diabetes is a disease in which glucose easily accumulates in the blood. Patients will have symptoms such as polyphagia, polydipsia, polyuria and weight loss. Diabetes is divided into type 1 and type 2 diabetes. Type 1 diabetes is characterized by a lack of insulin secretion ability, requiring daily insulin injections. It occurs at a young age and is more common in children and adolescents. Type 2 diabetes is caused by the body's inability to effectively use insulin. It is common in middle-aged and elderly people, with a high incidence in obese people. It is often accompanied by high blood pressure, dyslipidemia, arteriosclerosis and other diseases. If diabetes is not well controlled, it will cause complications, leading to failure and lesions in the kidneys, eyes, feet and other parts, and it cannot be cured.

[0003] α-glucosidase is an intestinal cell membrane enzyme whose main physiological function is to hydrolyze polysaccharides. Therefore, controlling the activity of α-glucosidase is an effective method for treating prediabetic patients and relieving diabetic symptoms. At present, some synthetic α-glucosidase inhibitors, such as acarbose and miglitol, have been used in clinical treatment, but these synthetic drugs can cause side effects such as digestive system disorders, intestinal flatulence, and liver poisoning. At present, some scholars have focused on some natural products in food, fruits and plants. These natural products have relatively small side effects and can achieve the purpose of controlling blood sugar in daily diet.

[0004] At present, diabetes is mainly treated with Western medicine, mainly chemical drugs such as insulin secretagogues, α-glucosidase inhibitors, insulin sensitizers, and insulin, such as sulfonylureas, biguanides, thiazolidinediones, etc. Although these chemical hypoglycemic drugs can effectively control the patient's blood sugar level, they will produce certain side effects after long-term use, which may further aggravate organ damage and even cause drug resistance, and the administration method also has many inconveniences. At the same time, these drugs have no ideal control effect on the early stage and chronic complications of diabetes, are prone to hypoglycemia or lactic acidosis, and cannot prevent further necrosis of pancreatic β cells, leading to further insulin dependence and other side effects.

[0005] According to traditional Chinese medicine, the basic pathogenesis of diabetes is yin deficiency and dryness and heat. Yin deficiency is the root cause, and dryness and heat are the symptoms. The two are mutually causal, and the affected organs are the lungs, spleen, and kidneys. In the early stage, yin deficiency leads to hyperactivity of fire, in the middle stage, qi is damaged and both qi and yin are deficient, and in the late stage, both yin and yang are deficient. Blood stasis may occur during the course of the disease; kidney yin is insufficient, the liver loses nourishment, and the eyes are not nourished, which may lead to dry eyes, blurred vision, and even blindness; internal stagnation of heat may cause sores, carbuncles, and ulcers; yin deficiency and dryness and heat, refining liquid into phlegm, may cause stroke and hemiplegia; kidney yin is insufficient, yin damages yang, spleen and kidney yang decline, water and moisture overflow, and become edema; extreme depletion of yin fluid leads to yin exhaustion and yang death, and there are dangerous symptoms such as unclear consciousness, dry skin, cold limbs, and weak pulse.

[0006] Chinese herbal medicine plays a significant role in preventing and treating various chronic complications of diabetes. New measures such as medicated diet have played a role in the health care effect of traditional Chinese medicine. In order to improve the disease through long-term quantitative health care, it is very valuable to develop auxiliary blood sugar lowering health care products. According to the characteristics of diabetes in traditional Chinese medicine, people have developed single or compound Chinese medicine preparations to treat diabetes, but the blood sugar lowering effect is not ideal.

[0007] The present invention optimizes the combination of several Chinese medicinal herbs, namely, mulberry, polygonatum, and eucommia male flower, based on an in-depth study of a large number of ancient prescriptions and combined with modern pharmaceutical research literature, in order to achieve a blood sugar lowering effect by giving full play to the synergistic effect of each raw material in lowering blood sugar. SUMMARY OF THE INVENTION

[0008] In order to solve the above existing problems, the present invention prepares a hypoglycemic pharmaceutical composition by selecting suitable botanical medicinal ingredients and their proportions. The hypoglycemic composition of the present invention has a clear hypoglycemic function, is safe, has no toxic side effects, and has a significant effect.

[0009] To this end, the present invention adopts the following technical solutions:

[0010] The invention provides a medicine composition with blood sugar lowering effect, which is composed of the following raw materials: mulberry, polygonatum sibiricum and eucommia male flower.

[0011] Furthermore, the mulberry is extracted mulberry polypeptide.

[0012] Furthermore, the preparation method of the mulberry polypeptide comprises the following steps:

[0013] (1) Pretreatment step: taking dry mulberries, washing to remove impurities, drying, and crushing to obtain mulberry powder;

[0014] (2) Homogenization: Soak in room temperature purified water, then add 1% cellulase by mass, adjust the pH value to 5.0, and hydrolyze at 40° C. for 2 h; then heat to keep warm, pour into a colloid mill while hot to homogenize to obtain a homogenous liquid, cool to room temperature and set aside;

[0015] (3) First enzymatic hydrolysis: adjust the pH value of the homogenate to 7.5, add enzyme, stir and react at 45°C for 1 h, heat to keep warm after hydrolysis, and obtain the first enzymatic hydrolyzate;

[0016] (4) Second enzymatic hydrolysis: The pH value of the first enzymatic hydrolyzate was adjusted to 10, enzyme was added, stirred and reacted at 40°C for 1 h, and heated to keep warm after hydrolysis to obtain the second enzymatic hydrolyzate;

[0017] (5) Polypeptide purification: The second enzymatic hydrolyzate is centrifuged to obtain a supernatant; then, the supernatant is ultrafiltered through a 3000Da ultrafiltration membrane to obtain a mulberry polypeptide with a molecular weight of less than 3000Da, which is then spray-dried into a powder for storage.

[0018] Furthermore, the enzyme used in the first enzymolysis is papain added at an amount of 20000 U / g.

[0019] Furthermore, the enzyme for the second enzymolysis is alkaline protease added at an addition amount of 40000U / g.

[0020] Furthermore, the pharmaceutical composition is composed of the following raw materials in parts by weight: 2-5 parts of mulberry polypeptide, 5-10 parts of polygonatum and 5-10 parts of eucommia male flowers.

[0021] The present invention also provides a method for preparing the pharmaceutical composition, comprising the following steps:

[0022] (1) taking Polygonatum sibiricum and Eucommia ulmoides male flowers, crushing them, sieving them, and mixing them evenly to obtain a Chinese medicine mixed powder;

[0023] (2) adding water to the mixed powder of traditional Chinese medicine and performing ultrasonic treatment;

[0024] (3) After the ultrasonic treatment is completed, decocting is performed to obtain a decoction;

[0025] (4) After the decoction is cooled to room temperature, microorganism A is added for fermentation; then microorganism B is added for secondary fermentation. After the fermentation is completed, the fermentation mixture is heated and inactivated to obtain a Chinese medicine fermentation liquid.

[0026] (5) filtering the Chinese medicine fermentation liquid, vacuum freeze-drying the filtrate into powder, then adding mulberry polypeptide, and mixing evenly to obtain a hypoglycemic drug composition.

[0027] Furthermore, the amount of water added during decoction in step (3) is 10-20 times the weight of the mixed powder, the water is added multiple times, and the decoction time is 1-5 hours.

[0028] Furthermore, the microorganism A is 2-5% by volume of Lactobacillus acidophilus ATCC4356.

[0029] Furthermore, the microorganism B is 2-5% by volume of Bacillus subtilis ATCC6633.

[0030] The present invention also provides the use of the pharmaceutical composition in preparing drugs for lowering blood sugar and / or treating diabetes.

[0031] Furthermore, the dosage forms of the drug include tablets, capsules, granules, pills, and oral liquid preparations.

[0032] Polygonatum sibiricum is a perennial herbaceous plant of the genus Polygonatum in the Liliaceae family, and its rhizome is used for medicinal purposes. The main varieties are Polygonatum sibiricum, Yunnan Polygonatum sibiricum and Polygonatum sibiricum, which are mainly found in the mountainous areas and hills in the central and western parts of my country. According to their shapes, they are commonly known as large Polygonatum sibiricum, chicken-head Polygonatum sibiricum, and turmeric Polygonatum sibiricum. The ones with large pieces, plumpness, yellow-white color, and transparent cross-section are superior, and those with bitter taste cannot be used as medicine. Polygonatum sibiricum is used for both medicine and food, and has always been a precious Chinese medicinal material. Its main ingredients include: Polygonatum sibiricum polysaccharides, steroidal saponins, anthraquinone compounds, alkaloids, cardiac glycosides, lignans, multiple vitamins, trace elements and amino acids, and its medicinal and nutritional values ​​are extremely high. Polygonatum sibiricum is dry in taste and flat in nature. It enters the lung, spleen, and kidney meridians, and has the effects of nourishing qi and yin; moistening the lungs and spleen, and nourishing the kidneys and filling essence. It is mainly used to treat cough due to yin deficiency, cough due to lung dryness, weakness due to spleen deficiency, poor appetite and dry mouth, weakness of waist and knees due to kidney deficiency, impotence and spermatorrhea, tinnitus and blurred vision, premature graying of hair and beard, emaciation due to physical weakness, wind leprosy and tinea, etc. It is mainly used to treat dry cough due to lung dryness, weakness due to physical weakness, palpitations and shortness of breath, dry mouth due to long-term illness, diabetes, hypertension, etc. It has good effects in antibacterial, anti-inflammatory, detoxification, anti-fatigue and anti-aging.

[0033] "Shennong's Herbal Classic" records that Eucommia ulmoides is mainly used to treat back pain, and has the effects of tonifying the middle, replenishing the essence, and strengthening the tendons and bones. Long-term use can make the body light and resist aging. Scientific research has found that Eucommia ulmoides male flowers contain more than 60 effective plant ingredients. Eucommia ulmoides male flowers bring together the essence of Eucommia ulmoides, and have all the health benefits of Eucommia ulmoides. In addition, they are particularly effective in "strengthening the liver, tonifying the kidneys, facilitating bowel movements, promoting sleep, and lowering the three highs". Eucommia ulmoides male flowers have been approved by the Ministry of Health as a new food raw material. Eucommia ulmoides male flowers contain active ingredients such as chlorogenic acid and quercetin, which have a significant effect on lowering blood sugar.

[0034] Mulberry is the fruit of the mulberry tree of the Moraceae family. Its aliases include mulberry seeds, mulberry sugarcane, mulberry dates, mulberry fruits, mulberry bubbles, and black mulberries. Compendium of Materia Medica records that "drinking mulberry juice can relieve alcohol poisoning, promote diuresis, reduce swelling, nourish yin and blood, and is used for liver and kidney deficiency, deficiency of essence and blood, dizziness, blurred vision, tinnitus and insomnia, and premature graying of hair." It is cold in nature, sweet and sour in taste, and enters the heart, liver, and kidney meridians. It has the effects of nourishing yin and blood, moistening the intestines, promoting the production of body fluids, and regulating immunity; it is used to regulate blood sugar, urine sugar, yin deficiency and blood deficiency, yin deficiency and thirst, body fluid deficiency and thirst, and dizziness and tinnitus. Mulberries are rich in vitamins and carotene, as well as linoleic acid, stearic acid, oleic acid, etc., which can prevent human arteriosclerosis, bone joint sclerosis, promote metabolism, promote the growth of red blood cells, prevent leukocytopenia, and have auxiliary effects on the treatment of diabetes, hypertension, hyperlipidemia and other diseases; and have good effects on neurasthenia, internal heat and thirst, deafness and blurred vision, blood deficiency and constipation.

[0035] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0036] (1) The present invention optimizes the formula based on traditional Chinese medicine theory and combines it with modern medical technology, screens multiple medicinal and edible plants and determines that mulberry, polygonatum and eucommia ulmoides corsage are the composition of the formula, and obtains the following raw material composition by weight: 2-5 parts of mulberry polypeptide, 5-10 parts of polygonatum and 5-10 parts of eucommia ulmoides corsage, so as to achieve the purpose of lowering blood sugar or treating diabetes, and has broad market prospects.

[0037] (2) The present invention uses specific Lactobacillus acidophilus and Bacillus subtilis to ferment the raw materials Polygonatum sibiricum and Eucommia ulmoides male flowers, promotes the generation of active products and the conversion of active ingredients, and synergistically inhibits the action of enzymes such as α-glucosidase in the intestine with mulberry polypeptide to regulate the blood sugar concentration in the body, making the drug effect stronger.

[0038] (3) The hypoglycemic pharmaceutical composition of the present invention uses Chinese medicine that is both medicinal and edible as the main raw material, is highly safe, has no toxic side effects, can be taken for a long time, effectively avoids the risk of liver damage caused by the co-use of multiple Chinese medicine components, and improves the safety of the drug and the compliance of patients. The preparation method of the present invention is simple in process, low in cost, and suitable for large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 . A graph showing the test results of the inhibition rate of α-glucosidase by the hypoglycemic compositions of the embodiments of the present invention and the comparative examples. Example

[0040] The present invention is further described below in conjunction with specific examples. Unless otherwise specified, the raw materials, reagents or devices used in the following examples can be obtained from conventional commercial sources or can be obtained by existing known methods.

[0041] It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturers.

[0042] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0043] In the specific embodiments of the present invention, mulberry, polygonatum sibiricum and eucommia male flower are all in dry weight and purchased from the market.

[0044] The strains of the present invention are all purchased from Shanghai Shifeng Biotechnology Co., Ltd., including: Lactobacillus acidophilus ATCC4356, Lactobacillus plantarum CGMCC1.557, Bacillus subtilis ATCC6633, Lactobacillus casei LWCC3002, Lactobacillus rhamnosus ATCC7469, and Bacillus licheniformis ATCC11946.

[0045] Preparation of basic example mulberry polypeptide

[0046] The preparation method of mulberry polypeptide comprises:

[0047] (1) Pretreatment step: taking dry mulberries, washing to remove impurities, drying, and crushing to 10-30 mesh to obtain mulberry powder;

[0048] (2) Homogenization: add room temperature pure water at a solid-liquid ratio of 1:1 (M / V) and soak for 60 minutes, then add cellulase at a mass ratio of 1%, adjust the pH value to 5.0, and hydrolyze at 40°C for 2 hours; then heat to 90°C and keep warm for 15 minutes, pour into a colloid mill while hot and homogenize for 1 minute to obtain a homogenous solution, cool to room temperature and set aside.

[0049] (3) First enzymatic hydrolysis: The pH value of the homogenate was adjusted to 7.5, and papain was added at an amount of 20,000 U / g. The mixture was stirred and reacted at 45°C for 1 h. After hydrolysis, the mixture was heated to 90°C and kept warm for 15 min to obtain the first enzymatic hydrolyzate.

[0050] (4) Second enzymatic hydrolysis: The pH value of the first enzymatic hydrolyzate was adjusted to 10, and alkaline protease was added at an amount of 40,000 U / g. The mixture was stirred and reacted at 40°C for 1 h. After hydrolysis, it was heated to 90°C and kept warm for 15 min to obtain the second enzymatic hydrolyzate.

[0051] (5) Polypeptide purification: The second enzymatic hydrolyzate was centrifuged at 8000 rpm for 10 min to obtain the supernatant; the supernatant was then ultrafiltered through a 3000 Da ultrafiltration membrane to obtain mulberry polypeptides with a molecular weight of less than 3000 Da, which were then spray-dried into powder for storage.

[0052] Example 1

[0053] A plant component hypoglycemic composition comprises the following components in parts by weight: 2 parts of mulberry polypeptide, 6 parts of polygonatum sibiricum and 10 parts of eucommia male flowers.

[0054] The specific preparation method comprises the following steps:

[0055] (1) Take 6 parts of polygonatum and 10 parts of eucommia male flowers, grind them, sieve them through 200 mesh, and mix them evenly to obtain a Chinese medicine mixed powder;

[0056] (2) Add 5 times the amount of water to the mixed powder of Chinese medicine and perform ultrasound for 30 minutes;

[0057] (3) After the ultrasonic treatment, decoction is performed. The amount of water added during decoction is 20 times the weight of the mixed powder. Water is added multiple times. The decoction time is 3 hours to obtain a decoction liquid.

[0058] (4) After the decoction is cooled to room temperature, Lactobacillus acidophilus (ATCC4356) with an inoculum amount of 3% by volume is added, and the mixture is fermented at a constant temperature of 37° C. for 16 hours; then, Bacillus subtilis (ATCC6633) with an inoculum amount of 4% by volume is added for secondary fermentation, and the mixture is fermented at 37° C. for 24 hours. After the fermentation is completed, the fermentation mixture is heated at 110° C. for 15 minutes for inactivation treatment to obtain a Chinese medicine fermentation liquid.

[0059] (5) Filter the Chinese medicine fermentation liquid, freeze-dry the filtrate in vacuum to form powder, then add 2 parts of mulberry polypeptide, mix well, and obtain a hypoglycemic composition.

[0060] Example 2

[0061] A plant component hypoglycemic composition comprises the following components in parts by weight: 3 parts of mulberry polypeptide, 10 parts of polygonatum sibiricum and 5 parts of eucommia male flowers.

[0062] The specific preparation method comprises the following steps:

[0063] (1) Take 10 parts of polygonatum and 5 parts of eucommia male flowers, grind them, sieve them through 200 mesh, and mix them evenly to obtain a Chinese medicine mixed powder;

[0064] (2) Add 5 times the amount of water to the mixed powder of Chinese medicine and perform ultrasound for 30 minutes;

[0065] (3) After the ultrasonic treatment, decoction is performed. The amount of water added during decoction is 15 times the weight of the mixed powder. Water is added multiple times. The decoction time is 4 hours to obtain a decoction liquid.

[0066] (4) After the decoction is cooled to room temperature, Lactobacillus acidophilus (ATCC4356) with an inoculum amount of 5% by volume is added, and the mixture is fermented at a constant temperature of 37° C. for 20 hours; then, Bacillus subtilis (ATCC6633) with an inoculum amount of 2% by volume is added for secondary fermentation, and the mixture is fermented at 37° C. for 18 hours. After the fermentation is completed, the fermentation mixture is heated at 110° C. for 15 minutes for inactivation treatment to obtain a Chinese medicine fermentation liquid.

[0067] (5) Filter the Chinese medicine fermentation liquid, freeze-dry the filtrate in vacuum to form powder, then add 3 parts of mulberry polypeptide, mix well, and obtain a hypoglycemic composition.

[0068] Example 3

[0069] A plant component hypoglycemic composition comprises the following components in parts by weight: 4 parts of mulberry polypeptide, 8 parts of polygonatum sibiricum and 6 parts of eucommia male flowers.

[0070] The specific preparation method comprises the following steps:

[0071] (1) Take 8 parts of polygonatum and 6 parts of eucommia male flowers, grind them, sieve them through 200 mesh, and mix them evenly to obtain a Chinese medicine mixed powder;

[0072] (2) Add 5 times the amount of water to the mixed powder of Chinese medicine and perform ultrasound for 30 minutes;

[0073] (3) After the ultrasonic treatment, decoction is performed. The amount of water added during decoction is 10 times the weight of the mixed powder. Water is added multiple times. The decoction time is 1 hour to obtain a decoction liquid.

[0074] (4) After the decoction is cooled to room temperature, Lactobacillus acidophilus (ATCC4356) with an inoculum amount of 2% by volume is added, and the mixture is fermented at a constant temperature of 37°C for 24 hours; then, Bacillus subtilis (ATCC6633) with an inoculum amount of 5% by volume is added for secondary fermentation, and the mixture is fermented at 37°C for 12 hours. After the fermentation is completed, the fermentation mixture is heated at 110°C for 15 minutes for inactivation treatment to obtain a Chinese medicine fermentation liquid.

[0075] (5) Filter the Chinese medicine fermentation liquid, freeze-dry the filtrate in vacuum to form powder, then add 4 parts of mulberry polypeptide, mix well, and obtain a hypoglycemic composition.

[0076] Example 4

[0077] A plant component hypoglycemic composition comprises the following components in parts by weight: 5 parts of mulberry polypeptide, 5 parts of polygonatum sibiricum and 8 parts of eucommia male flowers.

[0078] The specific preparation method comprises the following steps:

[0079] (1) Take 5 parts of polygonatum and 8 parts of eucommia male flowers, grind them, sieve them through 200 mesh, and mix them evenly to obtain a Chinese medicine mixed powder;

[0080] (2) Add 5 times the amount of water to the mixed powder of Chinese medicine and perform ultrasound for 30 minutes;

[0081] (3) After the ultrasonic treatment, decoction is performed. The amount of water added during decoction is 15 times the weight of the mixed powder. Water is added multiple times. The decoction time is 5 hours to obtain a decoction liquid.

[0082] (4) After the decoction is cooled to room temperature, Lactobacillus acidophilus (ATCC4356) with an inoculum amount of 4% by volume is added, and the mixture is fermented at a constant temperature of 37°C for 16 hours; then Bacillus subtilis (ATCC6633) with an inoculum amount of 3% by volume is added for secondary fermentation, and the mixture is fermented at 37°C for 24 hours. After the fermentation is completed, the fermentation mixture is heated at 110°C for 15 minutes for inactivation treatment to obtain a Chinese medicine fermentation liquid.

[0083] (5) Filter the Chinese medicine fermentation liquid, freeze-dry the filtrate in vacuum to form powder, then add 5 parts of mulberry polypeptide, mix well, and obtain a hypoglycemic composition.

[0084] Comparative Example 1

[0085] Compared with Example 1, this comparative example directly uses mulberries for fermentation to prepare the hypoglycemic composition.

[0086] A plant component hypoglycemic composition comprises the following components in parts by weight: 2 parts of mulberry, 6 parts of polygonatum, and 10 parts of eucommia male flowers.

[0087] The specific preparation method comprises the following steps:

[0088] (1) Take 2 parts of mulberry, 6 parts of polygonatum and 10 parts of eucommia male flower, grind them, sieve them through 200 mesh, and mix them evenly to obtain a mixed Chinese medicine powder;

[0089] (2) Add 5 times the amount of water to the mixed powder of Chinese medicine and perform ultrasound for 30 minutes;

[0090] (3) After the ultrasonic treatment, decoction is performed. The amount of water added during decoction is 20 times the weight of the mixed powder. Water is added multiple times. The decoction time is 3 hours to obtain a decoction liquid.

[0091] (4) After the decoction is cooled to room temperature, Lactobacillus acidophilus (ATCC4356) with an inoculum amount of 3% by volume is added, and the mixture is fermented at a constant temperature of 37° C. for 16 hours; then, Bacillus subtilis (ATCC6633) with an inoculum amount of 4% by volume is added for secondary fermentation, and the mixture is fermented at 37° C. for 24 hours. After the fermentation is completed, the fermentation mixture is heated at 110° C. for 15 minutes for inactivation treatment to obtain a Chinese medicine fermentation liquid.

[0092] (5) filtering the Chinese medicine fermentation liquid, and vacuum freeze-drying the filtrate into powder to obtain a hypoglycemic composition.

[0093] Comparative Example 2

[0094] Compared with Example 1, the final composition of this comparative example does not contain mulberry polypeptide.

[0095] A plant component hypoglycemic composition comprises the following components in parts by weight: 6 parts of polygonatum sibiricum and 10 parts of eucommia male flowers.

[0096] The specific preparation method comprises the following steps:

[0097] (1) Take 6 parts of polygonatum and 10 parts of eucommia male flowers, grind them, sieve them through 200 mesh, and mix them evenly to obtain a Chinese medicine mixed powder;

[0098] (2) Add 5 times the amount of water to the mixed powder of Chinese medicine and perform ultrasound for 30 minutes;

[0099] (3) After the ultrasonic treatment, decoction is performed. The amount of water added during decoction is 20 times the weight of the mixed powder. Water is added multiple times. The decoction time is 3 hours to obtain a decoction liquid.

[0100] (4) After the decoction is cooled to room temperature, Lactobacillus acidophilus (ATCC4356) with an inoculum amount of 3% by volume is added, and the mixture is fermented at a constant temperature of 37° C. for 16 hours; then, Bacillus subtilis (ATCC6633) with an inoculum amount of 4% by volume is added for secondary fermentation, and the mixture is fermented at 37° C. for 24 hours. After the fermentation is completed, the fermentation mixture is heated at 110° C. for 15 minutes for inactivation treatment to obtain a Chinese medicine fermentation liquid.

[0101] (5) filtering the Chinese medicine fermentation liquid, and vacuum freeze-drying the filtrate into powder to obtain a hypoglycemic composition.

[0102] Comparative Example 3

[0103] A plant component hypoglycemic composition comprises the following components in parts by weight: 5 parts of mulberry polypeptide, 1 part of polygonatum sibiricum and 1 part of eucommia male flower.

[0104] The specific preparation method comprises the following steps:

[0105] (1) Take 1 part of polygonatum and 1 part of eucommia male flower, grind them, sieve them through 200 mesh, and mix them evenly to obtain a Chinese medicine mixed powder;

[0106] (2) Add 5 times the amount of water to the mixed powder of Chinese medicine and perform ultrasound for 30 minutes;

[0107] (3) After the ultrasonic treatment, decoction is performed. The amount of water added during decoction is 20 times the weight of the mixed powder. Water is added multiple times. The decoction time is 3 hours to obtain a decoction liquid.

[0108] (4) After the decoction is cooled to room temperature, Lactobacillus acidophilus (ATCC4356) with an inoculum amount of 3% by volume is added, and the mixture is fermented at a constant temperature of 37° C. for 16 hours; then, Bacillus subtilis (ATCC6633) with an inoculum amount of 4% by volume is added for secondary fermentation, and the mixture is fermented at 37° C. for 24 hours. After the fermentation is completed, the fermentation mixture is heated at 110° C. for 15 minutes for inactivation treatment to obtain a Chinese medicine fermentation liquid.

[0109] (5) Filter the Chinese medicine fermentation liquid, freeze-dry the filtrate in vacuum to form powder, then add 5 parts of mulberry polypeptide, mix well, and obtain a hypoglycemic composition.

[0110] Comparative Example 4

[0111] An equal amount of wolfberry was used to replace polygonatum, and the other raw materials and preparation method were exactly the same as those in Example 2.

[0112] Comparative Example 5

[0113] An equal amount of Astragalus was used to replace the male flowers of Eucommia ulmoides. The other raw materials and preparation method were exactly the same as those in Example 3.

[0114] Comparative Example 6

[0115] Lactobacillus casei LWCC3002 was used instead of Lactobacillus acidophilus ATCC4356, and the other raw materials and preparation method were exactly the same as those in Example 1.

[0116] Comparative Example 7

[0117] Bacillus licheniformis ATCC11946 was used to replace Bacillus subtilis ATCC6633, and the remaining raw materials and preparation method were exactly the same as those in Example 2.

[0118] Comparative Example 8

[0119] Lactobacillus rhamnosus ATCC7469 replaced Lactobacillus acidophilus ATCC4356, and Bacillus licheniformis ATCC11946 replaced Bacillus subtilis ATCC6633. The remaining raw materials and preparation method were exactly the same as those in Example 3.

[0120] Comparative Example 9

[0121] Lactobacillus plantarum CGMCC1.557 was used to replace Lactobacillus acidophilus ATCC4356, and Bacillus licheniformis ATCC11946 was used to replace Bacillus subtilis ATCC6633. The remaining raw materials and preparation method were exactly the same as those in Example 4.

[0122] Test Example 1 Inhibition test of hypoglycemic composition on glucose metabolism-related enzymes

[0123] α-Glucosidase belongs to the class of oligosaccharide hydrolases. Its inhibitors competitively inhibit the action of glycosidases on the villi of the small intestinal epithelium to reduce the degradation of sugars, delay the digestion and absorption of sugars, and thus effectively reduce the peak blood sugar concentration after meals in diabetics, achieving the purpose of controlling blood sugar. α-Glucosidase inhibitors (such as acarbose, etc.) have become a class of oral hypoglycemic drugs widely used in clinical practice.

[0124] The activity screening of α-glucosidase inhibitors can be detected by the in vitro enzymatic reaction between the enzyme and its substrate p-nitrophenyl-β-D-pyranogalactoside (PNPG, a maltose analogue). After α-glucosidase is added to the substrate of the enzyme reaction, the substrate is decomposed into p-nitrophenol (PNP) and glucose by the enzyme. PNP is a colored substance with a maximum absorption at around 405nm. It can be measured by an enzyme reader, and the inhibitory activity of the sample is calculated based on the OD value.

[0125] Test method:

[0126] (1) The experiment was divided into a blank group, a drug group (the hypoglycemic composition prepared in the examples and comparative examples) and a drug blank group (only the drug was added). Each reactant was added to a 96-well plate according to the dose, with 3 parallel wells in each group.

[0127] (2) Add 50 μL of 10 mg / L sample solution, 50 μL of 0.1 U / ml enzyme solution, 100 μL of 50 mmol / L PNPG substrate, and pH 7.0 phosphate buffer to a 96-well ELISA plate in sequence and mix thoroughly. Set up a blank control without drugs (add phosphate buffer). PNPG can be hydrolyzed to produce glucose and PNP under the action of α-glucosidase. Incubate at 37°C for 60 minutes and measure its absorbance at a wavelength of 405 nm. Calculate the inhibition rate of α-glucosidase activity. The inhibition rate calculation formula is as follows:

[0128]

[0129] Among them, A0 is the OD value of the blank control group, and A1 is the OD value of the sample group.

[0130] Test results:

[0131] The compositions of the examples of the present invention and the comparative examples can inhibit the activity of α-glucosidase, but the inhibition rates of α-glucosidase are different. Figure 1 As shown, the inhibition rates of the compositions in Examples 1 to 4 on α-glucosidase are 64.25%, 60.21%, 65.28% and 61.46%, respectively, all exceeding 50%; the inhibition rates of the compositions in Comparative Examples 1-9 on α-glucosidase are all lower than 50%, with the lowest rates of 28.65% and 25.80% in Comparative Example 5 and Comparative Example 7, respectively. In Comparative Example 5, Astragalus, which also has the function of lowering blood sugar, replaces Eucommia male flower, and its inhibitory effect after fermentation is far different from that of Example 3; and in Comparative Example 7, Bacillus licheniformis replaces Bacillus subtilis, and its inhibitory effect is nearly 35% different from that of Example 2.

[0132] It shows that the invention, after the extracts of polygonatum and eucommia male flowers fermented successively by Lactobacillus acidophilus and Bacillus subtilis are compounded with mulberry polypeptide in a certain proportion, has a significantly higher inhibitory effect on α-glucosidase than the preparation process of other groups, and there is a significant synergistic effect between mulberries, polygonatum, eucommia corsage and microorganisms, and has a significant blood sugar lowering effect.

[0133] Experimental Example 2 Therapeutic effect of hypoglycemic composition on hyperglycemic mice

[0134] (1) Test sample: the hypoglycemic composition prepared in Examples 1-4 and Comparative Examples 1-9.

[0135] (2) Animal grouping: SPF-grade SD rats, half male and half female, weighing 270 g ± 30 g, were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., with certificate number SCXK (Beijing) 2012-0001. After purchase, the animals were kept at an ambient temperature of (25 ± 1) °C with natural light, and fed with normal feed ad libitum, and allowed to acclimate to the environment for 7 days.

[0136] The mice were randomly divided into blank control group, model control group, positive control group, Example 1 to 4 groups, and Comparative Example 1 to 9 groups, with 160 mice in total, 10 mice in each group, half male and half female.

[0137] (3) Construction of T2DM (type 2 diabetes) rat model: Except for 10 mice in the blank control group, the other 150 rats were fed with a high-sugar diet for 4 weeks and then fasted for 12 hours without water deprivation. A single intraperitoneal injection of streptozotocin (STZ) 45 mg / kg was given, dissolved in a 1% concentration of citric acid buffer (pH 4.2-4.5) during injection. After 72 hours, blood was collected from the tail vein at the same time for 3 consecutive days to test the blood glucose level of rats in each group. If the blood glucose level was greater than 16 mmol / L for 3 tests, the T2DM model was successfully established.

[0138] (4) Administration: The Example group and the Comparative Example group were administered with the hypoglycemic composition prepared in Examples 1-4 and Comparative Examples 1-9 by gavage, once a day for 2 consecutive weeks, with a dosage of 0.5 g / kg, and a dosage volume of 0.1 mL / 10 g; the blank control group and the model control group were respectively administered with an equal amount of distilled water, once a day for 2 consecutive weeks; the positive control group was administered with 150 mg / (kg·d) metformin hydrochloride tablets (Harbin Labton Pharmaceutical Co., Ltd., batch number: National Medicine Standard H23021803) by gavage. During the administration period, the blank group rats were fed with ordinary feed, and the other groups were fed with high-sugar feed.

[0139] (5) Index measurement: After 2 weeks of drug administration, the rats in each group were fasted for 12 hours but not water, and then blood was collected by tail cutting to measure blood glucose levels. The test results are shown in Table 1.

[0140] Table 1 Effect of hypoglycemic composition on hypoglycemic effect

[0141]

[0142]

[0143] (Note: Compared with the blank group, ** indicates P < 0.01, * indicates P < 0.05; compared with the model control group, # Indicates P<

[0144] 0.05, ## Indicates P < 0.01.)

[0145] As can be seen from the model group in Table 1, the blood glucose value of the model rats before administration was 22.88±2.45mmol / L, which was significantly higher than that of the blank group, and there was a significant statistical difference between the two, indicating that the model was successfully established, and the other positive control group, embodiment group and comparative example group were also successfully established. After 2 weeks of administration, the positive control group, embodiment 1-4 groups and comparative example 1-9 groups were able to reduce the blood glucose concentration of the rats. In addition, the blood glucose values ​​of the positive control group and embodiment 1-4 groups were significantly different from those of the model control group (p<0.01), and compared with comparative examples 1-9, they had a more significant hypoglycemic effect, indicating that the hypoglycemic composition of the present invention has a good application prospect in the fields of medicine and health care products.

[0146] The above contents are further detailed descriptions of the present invention in combination with specific preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For ordinary technicians in the technical field to which the present invention belongs, several simple deductions or substitutions can be made without departing from the concept of the present invention, which should be regarded as falling within the protection scope of the present invention.

Claims

1. A pharmaceutical composition having a hypoglycemic effect, characterized in that: The invention is composed of the following raw materials: 2-5 parts of mulberry polypeptide, 5-10 parts of polygonatum sibiricum and 5-10 parts of eucommia male flower; The preparation method of the pharmaceutical composition comprises the following steps: (1) Take 5-10 parts of polygonatum and 5-10 parts of eucommia male flowers, crush them, sieve them, and mix them evenly to obtain a mixed Chinese medicine powder; (2) Add water to the mixed powder of traditional Chinese medicine and perform ultrasonic treatment; (3) After the ultrasonic treatment is completed, decoction is performed to obtain a decoction liquid; (4) After the decoction is cooled to room temperature, 2-5% by volume of Lactobacillus acidophilus ATCC4356 is added for fermentation; then 2-5% by volume of Bacillus subtilis ATCC6633 is added for secondary fermentation. After the fermentation is completed, the fermentation mixture is subjected to a heating inactivation treatment to obtain a Chinese medicine fermentation liquid; (5) Filter the fermented Chinese medicine liquid, freeze-dry the filtrate into powder by vacuum lyophilization, then add 2-5 parts of mulberry polypeptide, mix well, and obtain a drug composition with hypoglycemic effect.

2. The pharmaceutical composition according to claim 1, characterized in that The preparation method of the mulberry polypeptide comprises the following steps: (1) Pretreatment step: washing dry mulberries to remove impurities, drying them, and crushing them to obtain mulberry powder; (2) Homogenization: Soak in pure water at room temperature, then add 1% cellulase by mass, adjust the pH value to 5.0, and hydrolyze at 40°C for 2 hours; then heat to 90°C and keep warm, pour into a colloid mill while hot to obtain a homogenous solution, cool to room temperature and set aside; (3) First enzymatic hydrolysis: The pH value of the homogenate was adjusted to 7.5, and papain was added at a dosage of 20,000 U / g. The mixture was stirred at 45°C for 1 h. After hydrolysis, the mixture was heated to 90°C for preservation to obtain the first enzymatic hydrolyzate. (4) Second enzymatic hydrolysis: The pH value of the first enzymatic hydrolyzate was adjusted to 10, and alkaline protease was added at an amount of 40,000 U / g. The mixture was stirred at 40°C for 1 h. After hydrolysis, the mixture was heated to 90°C and kept warm to obtain the second enzymatic hydrolyzate. (5) Polypeptide purification: Centrifuge the second enzymatic hydrolysate to obtain the supernatant; then ultrafilter through a 3000Da ultrafiltration membrane to obtain mulberry polypeptides with a molecular weight of less than 3000Da, and spray dry them into powder for storage.

3. The pharmaceutical composition according to claim 1, characterized in that The amount of water added during decoction in step (3) is 10-20 times the weight of the mixed powder, and water is added multiple times. The decoction time is 1-5 hours.

4. Use of the pharmaceutical composition according to claim 1 in the preparation of drugs for lowering blood sugar and / or treating diabetes.

5. The use according to claim 4, characterized in that: The dosage forms of the medicine include tablets, capsules, granules, pills and oral liquid preparations.

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