Composition with hypoglycemic effect and preparation method and use thereof

Abstract

The present invention relates to a composition with hypoglycemic effect and its preparation method and use, belonging to the field of medicine or health care products. The present invention provides a composition with hypoglycemic effect, which is prepared from the following raw materials in weight ratio: 8-12 parts of black plum, 8-12 parts of astragalus, 8-12 parts of kudzu root, 2.4-3.6 parts of lotus seed heart, 5.6-8.4 parts of gardenia, 8-12 parts of wolfberry, 5.6-8.4 parts of yam, and 2.4-3.6 parts of American ginseng. The present invention also provides a preparation method and use of the composition. The present invention uses all medicines together, taking into account the upper, middle and lower three energizers, and together they have the effects of invigorating qi and nourishing yin, promoting body fluid and quenching thirst, nourishing yin and clearing heat. Compared with the single or simple drug pair in the literature, more synergistic effects may be produced between the components, and chronic endocrine system diabetes is comprehensively treated from multiple angles, and the condition is improved more comprehensively, and the synergistic effects between each other are utilized to enhance the efficacy, reduce adverse reactions, and have a wider scope of application.

Description

Technical Field

[0001] The invention relates to a composition with hypoglycemic effect and a preparation method and application thereof, belonging to the field of medicines or health products. Background Art

[0002] Diabetes is a global health challenge. According to data, the number of people suffering from diabetes worldwide has exceeded 500 million in 2021, and it is expected to affect 1.3 billion people by 2050. As the aging of the population continues to deepen, the number of elderly patients with diabetes is still on the rise. Western medicine usually treats diabetes by injecting insulin or taking oral hypoglycemic drugs such as biguanides, sulfonylureas, aldose reductase inhibitors, and α-glucosidase inhibitors. However, adverse reactions such as hypoglycemia, insulin allergy, diarrhea, stomach discomfort or colic may also occur during the use of the drug. The understanding and prevention of diabetes in traditional Chinese medicine was discussed as early as in the Yellow Emperor's Classic of Internal Medicine, and it plays a role in synergistic hypoglycemic, improving symptoms and signs, preventing and treating complications, improving quality of life and tertiary prevention.

[0003] There are many literatures reported on the treatment of type 2 diabetes, such as: Yu Li, et al., Effect and pharmacological analysis of Huangqi Wumei Decoction in the treatment of type 2 diabetes, Health Friend, January 2021, analyzed the pharmacological effects and clinical effects of Huangqi Wumei Decoction in the treatment of type 2 diabetes. Among them, Huangqi Wumei Decoction, 50-200g of Astragalus, 20-50g of Plum, was used. The experiment proved that taking Huangqi Wumei Decoction was effective and safe in the population of patients with type 2 diabetes. Wang Zuoke, et al., Analysis of the efficacy of Huangqi Wumei Decoction on type 2 diabetes with deficiency of both qi and yin, Diabetes New World, August 2020, explored the therapeutic effect of Huangqi Wumei Decoction in the treatment of type 2 diabetes with deficiency of both qi and yin. The dosage of Astragalus and Plum is selected according to the specific situation of the patient. The dosage of three groups of Astragalus and Plum is disclosed in the literature, namely: 50g of Astragalus and 15g of Plum; 100g of Astragalus and 25g of Plum; 200g of Astragalus and 50g of Plum. The application of Huangqi Wumei Decoction in the treatment of type 2 diabetes with deficiency of both qi and yin can achieve a more definite effect, help improve the patient's symptoms and signs, and is beneficial to blood sugar control, with few adverse reactions and high medication safety. Zhou Yingjie, et al., Exploration of the mechanism of action of American ginseng-astragalus in the treatment of type 2 diabetes based on network pharmacology and molecular docking technology, World Chinese Medicine, Vol. 18, No. 18, September 2023, explores the mechanism of action of American ginseng-astragalus in the treatment of type 2 diabetes (T2DM). It is disclosed that the mechanism of action of American ginseng-astragalus in the treatment of T2DM has the characteristics of multiple components, multiple targets, and multiple pathways. The document discloses that the American ginseng-astragalus drug pair is the main component of Yiqi Shengjin San, and the two are compatible to enhance the effect of invigorating qi and nourishing yin, but the document does not disclose the weight ratio of astragalus and American ginseng. Pan Yang et al., Effects of Lotus Seed Core and Nef on Experimental Diabetes and Obese Rat Models, Journal of Nanjing University of Chinese Medicine, Vol. 19, No. 4, July 2003, disclosed that Lotus Seed Core and Nef have certain effects on lowering blood sugar and regulating blood lipids in experimental diabetic and obese rats.

[0004] CN201710704155.7, invention name: A Chinese medicine for treating diabetes and controlling its complications, discloses a Chinese medicine for treating diabetes and controlling its complications, relates to a Chinese medicine, and specifically relates to a Chinese medicine for treating diabetes. The raw materials include the following weight ratios: American ginseng 33-37; astragalus 142-158; yam 95-105; black plum 76-84; Euryale ferox 67-73; sea buckthorn 100-110; schisandra chinensis 52-58; jujube seed 33-37; pumpkin powder 95-105; kudzu root 71-79; licorice 33-37; chromium gluconate 0.12-0.124. This patent requires the combination of traditional Chinese medicine and chemical compounds to achieve its effect. Among them, chromium gluconate is helpful for blood sugar control in patients with type 2 diabetes, but it has obvious side effects. Some people may experience gastrointestinal reactions such as nausea, vomiting, diarrhea, abdominal pain, etc., which may cause headaches, insomnia, mood changes, inattention and other problems. Long-term or excessive intake of chromium may cause liver and kidney damage. The chromium may interact with drugs such as insulin and thyroid hormones, affecting the efficacy of the drugs.

[0005] The above literature focuses on Astragalus-Prunus mume, American ginseng-Astragalus and lotus seed core respectively, and shows the therapeutic effect on diabetes from the perspective of combination and single medicine. However, one of the characteristics of compound Chinese medicine is that single medicine or simple medicine pair cannot fully adapt to the symptoms or the needs of the patient's condition. Summary of the invention

[0006] The invention provides a composition with hypoglycemic effect, a preparation method and application thereof.

[0007] The present invention provides a composition with hypoglycemic effect, which is prepared from the following raw materials in weight ratio:

[0008] 8-12 parts of black plum, 8-12 parts of astragalus, 8-12 parts of kudzu root, 2.4-3.6 parts of lotus seed core, 5.6-8.4 parts of gardenia, 8-12 parts of wolfberry, 5.6-8.4 parts of yam, and 2.4-3.6 parts of American ginseng.

[0009] Preferably, it is prepared from the following raw materials in weight ratio:

[0010] 10 parts of black plums, 10 parts of astragalus, 10 parts of kudzu root, 3 parts of lotus seed core, 7 parts of gardenia, 10 parts of wolfberries, 7 parts of yam, and 3 parts of American ginseng.

[0011] The composition of the present invention is prepared from the raw material powder and water extract as active ingredients, and is added with acceptable auxiliary materials or auxiliary ingredients to form a commonly used pharmaceutical preparation.

[0012] Wherein, the preparation is granules, capsules, pills, oral liquids, tablets.

[0013] The present invention also provides a method for preparing the composition having a hypoglycemic effect, which comprises the following steps:

[0014] a. Weigh the raw materials of each weight ratio;

[0015] b. Grind into powder, or boil with water, filter, concentrate, and add acceptable excipients or auxiliary ingredients to prepare into pharmaceutical preparations commonly used.

[0016] Preferably, it comprises the steps of:

[0017] a. Weigh the raw materials of each weight ratio;

[0018] b. Add water and boil twice, 10 times the amount of water for the first time and 8 times the amount of water for the second time, and extract for 75 minutes each time; filter, combine the decoctions, and concentrate under reduced pressure at 60°C to an extract with a relative density of 1.218-1.264, add microcrystalline cellulose at a ratio of 3:2 of extract: auxiliary materials, mix well, dry, crush, add appropriate amount of 80% ethanol as a wetting agent, wet granulate, dry at 65°C, arrange, and package to obtain.

[0019] The invention provides use of the composition in preparing medicine for treating diabetes.

[0020] Wherein, the medicine is a medicine for treating type II diabetes.

[0021] The present invention also provides use of the composition in preparing a health product that helps maintain a healthy blood sugar level.

[0022] The prescription of the present invention consists of black plum, astragalus, kudzu root, lotus seed core, gardenia, wolfberry, yam and American ginseng. It is an empirical prescription that has been clinically used by the inventor for more than 20 years, has a definite efficacy, and is widely used in diabetic patients with chronic endocrine system diseases.

[0023] Traditional Chinese medicine believes that the basic pathogenesis of diabetes is yin deficiency as the root and dryness and heat as the symptom. The treatment of this disease is to invigorate qi and nourish yin, produce fluid and moisten dryness. Clinically, qi and yin deficiency syndrome is more common. In the prescription of the present invention, black plum produces fluid and quenches thirst, American ginseng greatly replenishes vital energy, invigorates qi and strengthens the spleen, quenches thirst and produces fluid. The two medicines are used together, and the sweet and sour yin is transformed, and the effect of invigorating qi and nourishing yin is played together, and they are the main medicine; Astragalus invigorates qi and raises yang, assists American ginseng in invigorating qi and consolidating the foundation, and Pueraria root produces fluid and quenches thirst, assists black plum in nourishing yin and producing fluid. The two medicines work together to invigorate qi and raise fluid, and are specially used to treat symptoms such as thirst and fatigue caused by qi and yin deficiency, and are the minister medicines; lotus seeds and gardenias are used to clear heat and remove dampness; wolfberry nourishes kidney yin, and yam invigorates qi and strengthens the spleen as the guiding medicine. All the medicines are used together, taking into account the upper, middle and lower three burners, and the effects of invigorating qi and nourishing yin, producing fluid and quenching thirst, and nourishing yin and clearing heat are played together. It is used for thirst caused by deficiency of both Qi and Yin, with symptoms of weight loss, thirst, polyphagia, polydipsia, and fatigue. It is also used for type II diabetes with the above symptoms.

[0024] The beneficial effects of the present invention are:

[0025] The drugs of the present invention are used together, taking into account the upper, middle and lower three burners, and together have the effects of invigorating qi and nourishing yin, promoting body fluid and quenching thirst, nourishing yin and clearing away heat. Compared with single or simple drug pairs in the literature, the various components may produce more synergistic effects, and comprehensively treat chronic endocrine system diabetes from multiple angles, improve the condition more comprehensively, utilize the synergistic effects between each other, enhance the therapeutic effect, reduce adverse reactions, and have a wider range of applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is the water absorption graph of the prescription;

[0027] Figure 2 This is the result of the investigation of decoction times (A: the effect of different decoction times on the paste yield; B: the effect of different decoction times on the content of index components);

[0028] Figure 3 This is the result of the decoction time investigation (A: the effect of different decoction times on the paste yield; B: the effect of different decoction times on the content of index components);

[0029] Figure 4 The results of the investigation of water addition (A: the effect of different water addition times on the paste yield; B: the effect of different water addition times on the content of index components;

[0030] Figure 5 The following are the response surface analysis results (A: water addition amount and extraction time; B: water addition amount and decoction times; C: extraction times and decoction time). DETAILED DESCRIPTION

[0031] Example 1 Preparation of hypoglycemic granules of the present invention

[0032] Weigh the API:

[0033] 60g in total: 10g black plum, 10g kudzu root, 10g astragalus, 10g wolfberry, 3.0g lotus seed core, 7.0g gardenia, 7.0g yam, 3.0g American ginseng;

[0034] According to the prescription, eight medicinal materials, including black plum, kudzu root, astragalus, gardenia, yam, wolfberry, American ginseng and lotus seed core, were weighed in proportion, and 8 times of water (10 times for the first time) were added, and the mixture was decocted twice, and each extraction was 75 minutes. Filter, combine the decoction, and concentrate under reduced pressure at 60°C to an extract with a relative density of 1.218~1.264, add microcrystalline cellulose at a ratio of 3:2 of extract: auxiliary material, mix well, dry, crush, add appropriate amount of 80% ethanol as a wetting agent, wet granulate, dry at 65°C, arrange, and package to obtain the product.

[0035] Example 2 Preparation of the hypoglycemic oral liquid of the present invention

[0036] Weigh the following raw materials: ebony 8 g, astragalus 8 g, kudzu root 8 g, lotus seed core 2.4 g, gardenia 5.6 g, wolfberry 8 g, yam 5.6 g, American ginseng 2.4 g; extract and concentrate according to the method of Example 1 to prepare an oral solution.

[0037] Example 3 Preparation of hypoglycemic granules of the present invention

[0038] The raw materials were weighed: 12 g of black plum, 12 g of astragalus, 12 g of kudzu root, 3.6 g of lotus seed core, 8.4 g of gardenia, 12 g of wolfberry, 8.4 g of yam, and 3.6 g of American ginseng, and granules were prepared according to the method of Example 1.

[0039] Example 4 Study on the process of the hypoglycemic granules of the present invention

[0040] 1 Test materials

[0041] 1.1 Instruments

[0042] Thermo U3000 high performance liquid chromatograph, Thermo Corporation, USA; PS-80A ultrasonic cleaner, Dongguan Jiekang Ultrasonic Equipment Co., Ltd.; RC30002 electronic balance, Chengdu Besec Instruments and Meters Research Institute; SQP analytical balance, Sartorius Company, Germany.

[0043] 1.2 Drug testing

[0044] Reference substances puerarin (batch number 23101302, mass fraction 98.10%), citric acid (batch number 22030307, ​​mass fraction 99.80%), ginsenoside Rb1 (batch number 21101405, mass fraction 99.51%), ursolic acid (batch number 22091604, mass fraction 98.53%), and ginsenoside Re (batch number 23071802, mass fraction 99.97%) were all purchased from Chengdu Glip Biotechnology Co., Ltd.

[0045] The prescription pieces of Wumei (batch number 230101), Pueraria root (batch number 230801), lotus seed heart (batch number 230801), Astragalus (batch number 231002), Gardenia (batch number 221002), Lycium barbarum (batch number 230901), American ginseng (batch number 230801), and Chinese yam (batch number 230901) were all provided by Sichuan Guoqiang Chinese Medicine Piece Company. The prescription excipient is microcrystalline cellulose (batch number 2024011201), which was provided by Chengdu Kelong Chemical Co., Ltd.

[0046] Acetonitrile (chromatographic grade, Fisher Scientific), purified water (China Resources C'estbon Beverage Co., Ltd.), phosphoric acid (Fisher Scientific), methanol (Sinopharm Chemical Reagent Co., Ltd.).

[0047] 2 Extraction process research

[0048] This prescription is an empirical prescription summarized based on clinical basis, and has been proven to have significant efficacy through many years of clinical practice. It is mainly presented in the form of decoction in clinical application. To ensure the efficacy, it is still extracted by water decoction. Design the process route and study the extraction process of hypoglycemic granules. Taking the main representative ingredients of the prescription, citric acid and puerarin, as well as the paste yield and the transfer rate of citric acid and puerarin, as the evaluation indicators, the orthogonal test method is used to optimize the extraction process.

[0049] 2.1 Determination of paste yield

[0050] Concentrate 200 mL of the Jiangtang granule extract under reduced pressure, evaporate to dryness in a water bath (105°C), and place under reduced pressure drying at 80°C until constant weight, thereby obtaining the solid content in the Jiangtang extract.

[0051] Extraction rate (%) = solid content in extract / weight of crude drug corresponding to extract × 100

[0052] 2.2 Selection of indicator components

[0053] Taking citric acid in black plum and puerarin in pueraria as evaluation indicators, the reasons for optimizing the process parameters are as follows:

[0054] (1) The main drug of Jiangtang Granule is Wumei, and the auxiliary drug is Pueraria root. Therefore, the active ingredients in Wumei and Pueraria root specified in the pharmacopoeia were used as evaluation indicators to examine the transfer rate of the active ingredients in Wumei and Pueraria root;

[0055] (2) Citric acid, the main active ingredient in black plum, is an organic acid with good water solubility. It has the function of acid-sweetening yin and assisting in the treatment of diabetes and can be used as an evaluation indicator for water extraction technology.

[0056] (3) Puerarin, the main active ingredient in Pueraria root, is an isoflavone component that is easily soluble in methanol and ethanol and slightly soluble in water. It has the effect of promoting the production of body fluids and quenching thirst. It can work with black plum to nourish yin and promote the production of body fluids to treat diabetes. It can be used as an evaluation indicator for water extraction technology.

[0057] 2.3 Determination of citric acid content

[0058] 2.3.1 Chromatographic conditions

[0059] The chromatographic column was Kromasil100-5 C18 (4.6 mm × 250 mm, 5 μm), the mobile phase was 0.5% ammonium dihydrogen phosphate aqueous solution (adjusted to pH = 3 with phosphoric acid) (A)-methanol (C), the detection wavelength was 220 nm, the column temperature was 30 °C, and the flow rate was 1.0 mL / min

[0060] Table 1 Gradient elution table

[0061]

[0062] 2.3.2 Preparation of citric acid and puerarin reference solutions

[0063] Place the accurately weighed citric acid reference substance in a volumetric flask, dissolve and dilute with appropriate amount of pure water to prepare a 1.06 mg / mL reference substance solution; place the accurately weighed puerarin reference substance in a volumetric flask, dissolve and dilute with appropriate amount of methanol to prepare a 0.496 mg / mL reference substance solution.

[0064] 2.3.3 Preparation of test solution

[0065] Take 60g of ebony root 10g, kudzu root 10g, astragalus 10g, wolfberry 10g, lotus seed heart 3.0g, gardenia 7.0g, yam 7.0g, American ginseng 3.0g, and boil with water as the extraction solvent. Extraction was performed using Box-Behnken response surface design combination, and the compound decoction was filtered with 250 mesh gauze, which was concentrated under reduced pressure to a fluid extract with a relative density of 1.218~1.264. Take 1.0g of the fluid extract, add 30ml of pure water to dissolve, ultrasonicate for 30min, and filter through a 0.22 μm filter membrane to obtain the test solution.

[0066] 2.4 Determination of water absorption rate of medicinal materials

[0067] Weigh eight medicinal materials including black plum according to the prescription ratio, add 500 ml of water to soak the medicinal materials, filter and measure the filtered water every 0.5 h, and filter out the remaining water after 2 h. The water absorption rate was measured to be 127.8%. Therefore, during the experiment, 1.5 times the total amount of water of the medicinal materials should be added for the first decoction to prevent experimental errors caused by water absorption. Results Figure 1.

[0068] 2.5 Water extraction factor investigation

[0069] The hypoglycemic granules are composed of black plum, American ginseng, astragalus, kudzu root, lotus seed core, gardenia, wolfberry and yam in a certain proportion. By fixing other influencing factors and changing a single influencing factor, the influence of a single factor on the entire experimental process is verified, and then three factor levels are selected, on this basis, the extraction process is optimized. The number of decoctions, the amount of water added, and the decoction time are used as the investigation factors, and the citric acid in the prescription black plum, the puerarin in the pueraria, the citric acid conversion rate, the puerarin conversion rate and the paste yield rate are used as evaluation indicators for investigation.

[0070] 2.5.1 Decoction frequency investigation

[0071] Weigh 5 portions of slices according to the ratio, add water at a ratio of 1:8, add twice the amount of water for the first decoction, decoct and extract 1, 2, 3, 4, and 5 times, the extraction time is 1 hour, filter with silk cloth, prepare the test solution, calculate the paste rate and the content of each index component, and the change curve of the paste rate and the content of each index component with the number of extractions is shown in Figure 2 Therefore, the final number of extractions was 2, 3, and 4.

[0072] 2.5.2 Decoction time study

[0073] Weigh 5 portions of slices according to the ratio, add water at a ratio of 1:8, add twice the amount of water for the first decoction, decoct and extract 3 times, the extraction time is 30 min, 45 min, 60 min, 75 min, 90 min, respectively, filter with silk cloth, prepare the test solution, calculate the paste rate and the content of each index component, the paste rate and the content of each index component change with extraction time curve is shown in Figure 3 Therefore, the final extraction time was selected as 45 min, 60 min, and 75 min.

[0074] 2.5.3 Water addition times

[0075] Weigh 5 portions of slices and add water at 1:6, 1:8, 1:10, 1:12, and 1:14, respectively. Add twice as much water for the first decoction. Decoction and extraction are performed 3 times, each time for 60 min. After filtering with silk cloth, the test solution is prepared. Sample injection and determination are performed. The curves of the paste yield and the content of the index components changing with the material-liquid ratio are shown in Figure 2. Figure 4 . Finally, 1:8, 1:10, and 1:12 were selected.

[0076] 2.6 Determination of weights of citric acid, puerarin, dry extract rate, citric acid transfer rate, and puerarin transfer rate

[0077] In the study of the extraction process of hypoglycemic granules, citric acid, puerarin, dry extract rate, citric acid transfer rate and puerarin transfer rate were used as evaluation indicators. The weights of relevant indicators were determined by scientific methods in order to systematically evaluate the water extraction process.

[0078] In weight allocation, commonly used scientific methods include the analytic hierarchy process (AHP) and the index-related weight assignment method (CRITIC). AHP decomposes the problem into three layers, namely the target layer, the index layer, and the solution layer, constructs a judgment matrix, and solves the weights through the eigenvalue method to perform consistency tests. The advantage of AHP is that it combines qualitative and quantitative analysis and is suitable for complex multi-objective problems. CRITIC determines the importance of each indicator to the model performance by analyzing the correlation of multiple indicators of model performance. Its advantage is that it comprehensively considers the comparison strength and conflict of indicators, and gives higher weights to indicators with high information content and independence.

[0079] In traditional Chinese medicine prescriptions, monarch, minister, assistant and envoy are the basic principles of prescription composition, which is a high-level summary of the medication rules of prescriptions. It discusses the status of each drug in the prescription and the change rules of the properties and effects after compatibility from a multi-dimensional perspective. Therefore, it is not possible to simply define an indicator as "large information volume and high independence" and use the weight analysis method based on indicator correlation (CRITIC). In order to reflect the compatibility relationship of the monarch, minister, assistant and envoy, treatment method and prescription of the prescription, the hierarchical analysis method (AHP) is proposed to comprehensively evaluate the five related indicators involved in the extraction process of the hypoglycemic granules, as shown in Table 3. According to the compatibility relationship of the drugs in the prescription, the pharmacological effects of each indicator, the impact on the production process of granules and many other factors, the following priorities are determined: citric acid> puerarin> citric acid transfer rate = puerarin transfer rate> dry paste rate, and the 5-order judgment matrix is ​​scaled accordingly, with citric acid marked as 4, puerarin marked as 3, citric acid conversion rate and puerarin conversion rate both marked as 2, and dry paste rate marked as 1. The results of constructing a five-order judgment matrix for pairwise comparison based on this scale are shown in Table 2.

[0080] Table 2 Results of the 5th-order judgment matrix

[0081]

[0082] Table 3 Results of AHP hierarchical analysis

[0083]

[0084] The weight values ​​based on the constructed 5th-order judgment matrix were as follows: 33.77% for citric acid, 25.33% for puerarin, 16.89% for citric acid transfer rate, 15.57% for puerarin transfer rate, and 8.44% for dry cream.

[0085] Consistency test is a key step in AHP. The consistency of the results is evaluated by the consistency ratio (CR) value. The calculation formula is: CR = CI / RI

[0086] The consistency ratio (CR value) is used to evaluate the consistency of the matrix. Generally, if CR < 0.1, the judgment matrix is ​​considered to meet the consistency standard. If CR ≥ 0.1, the judgment matrix needs to be modified.

[0087] In the consistency test, the CI value is used to evaluate the consistency of the matrix, that is, whether the factors are logically smooth. The smaller the CI value, the more consistent the matrix. The calculation formula is as follows:

[0088] CI = λmax-n / n-1

[0089] Where λmax is the maximum eigenvalue of the judgment matrix, and n is the order of the judgment matrix.

[0090] The RI value in the consistency test refers to the random consistency index, which is used to evaluate the consistency of the judgment matrix. The RI value is related to the matrix order. The corresponding value can be found in Table 4:

[0091] Table 4 Random consistency RI table

[0092]

[0093] By calculation, we get λmax to be 5.383162, n is 5, and the CI value is 0.095 when substituted into the CI formula. The RI value of the 5th-order judgment matrix is ​​1.46 when checked in the RI table. Substituting the CI and RI values ​​into the calculation formula of the CR value, the CR value is 0.086. It can be concluded that the judgment matrix meets the consistency requirements and effectively determines the weights of the five indicators.

[0094] 2.7 Water extraction process optimization

[0095] 2.7.1 Water extraction process optimization - orthogonal experimental design Box-Behnken response surface optimization

[0096] The traditional decoction parameters of Chinese herbal medicine were used as indicators and the factor level was amplified in the literature to comprehensively design the orthogonal experimental factor level. The main parameters related to the extraction efficiency were the amount of water added (A), the total number of extractions (B), and the single extraction time (C). L9 (3 4 ) Orthogonal design was used to screen these three factors. The low, medium and high levels of the three factors were set as follows: decoction times A (8, 10 and 12 times), water addition times B (2, 3 and 4 times), decoction time C (45, 60 and 75 min). Indicators: dry extract weight, citric acid and puerarin content in dry extract, citric acid conversion rate, puerarin conversion rate, see Table 5.

[0097] The above-mentioned factor level table was used for the experiment. The dry paste rate was determined by the weighing method, the content of citric acid and puerarin in the solution was determined by HPLC, and the transfer rate of the index components of citric acid and puerarin was calculated. The results are shown in Table 6.

[0098] Table 5 Experimental arrangement of Box-Behnken response surface optimization for water extraction of sugar-reducing granules

[0099]

[0100] Table 6 Results of Box-Behnken response surface optimization test on water extraction process of Jiangtang prescription

[0101]

[0102] 2.7.2 Analysis of variance

[0103] The data of the sugar-reducing granules were fitted and analyzed by Design-Expert13, and a multiple regression equation was obtained between the score and the amount of water added (A), the number of decoctions (B), and the decoction time (C). The regression equation is: score = 84.81 + 2.08*A + 1.45*B + 7.20*C - 0.8934*AB + 1.56*AC - 2.27*BC + 2.06*A 2 -2.33*B 2 +0.3266*C 2 From the table, we can see that the model P <0.05, significant; lack of fit P =0.9040>0.05, R 2 =0.8891, indicating that the model has explanatory power.

[0104] According to the model, C has the most significant impact on the score, while A and B have no significant impact. As shown in Table 7, Figure 5 Therefore, the best solution is to use 8 times the amount of water and extract twice, each time for 75 minutes.

[0105] Table 7 Results of variance analysis

[0106]

[0107] 2.7.3 Best process verification test

[0108] To ensure the stability and reliability of the extraction process, the best process was used to extract the Jiangtang granules, and three parallel verification tests were conducted. The results are shown in Table 8. The verification results showed that the average comprehensive evaluation of the three verifications was 96.06, indicating that the model prediction results were reliable and the process had good reproducibility.

[0109] Table 8 Results of three batches of validation tests (n=3)

[0110]

[0111] 3. Investigation of Concentration Process

[0112] 3.1 Investigation of concentration methods

[0113] The water extract of Chinese herbal compound needs to be concentrated before it can be formed. Commonly used concentration methods are normal pressure (direct fire) and reduced pressure (rotary evaporation). These two methods are compared and analyzed. The decoction was concentrated to a relative density of 1.218~1.264 by normal pressure concentration and reduced pressure concentration (80℃, -0.1Mpa). The indexes were measured. The results showed that after reduced pressure concentration, all indicators were better than those of normal pressure concentration, as shown in Table 9.

[0114] Table 9 Results of investigation on concentration methods (n=2)

[0115]

[0116] 3.2 Concentration temperature investigation

[0117] During the vacuum concentration process, the choice of temperature has an important impact on the effective ingredients. Although too high a temperature will increase the rate, it will damage the heat-sensitive ingredients. Too low a temperature will prolong the heating time and reduce the concentration efficiency. Therefore, the selection of the concentration temperature plays a vital role.

[0118] The present invention comprehensively considers the effective ingredient retention and concentration efficiency. The decoction pieces are decocted according to the optimal extraction process, and the extracted medicinal liquid is concentrated to a relative density of 1.218-1.264 at 60°C, 70°C, and 80°C, and the index content is determined. The results are shown in Table 10. The score is the highest at 60°C, so 60°C is selected as the reduced pressure concentration temperature.

[0119] Table 10 Concentration temperature investigation results (n=3)

[0120]

[0121] 4 Molding process research

[0122] 4.1 Forming process selection and evaluation indicators

[0123] 4.1.1 Selection of molding process

[0124] In actual production, Chinese medicine granules are usually made by wet granulation, dry granulation, one-step granulation and rapid mixing granulation. Since hypoglycemic prescriptions are usually in the form of decoctions and do not contain precious fine medicinal materials, and considering production conditions and costs, the most commonly used wet granulation method is selected.

[0125] 4.1.2 Determination of forming rate

[0126] The granule test was carried out according to the double screening method in the 2020 edition of the Chinese Pharmacopoeia. The total mass of the hypoglycemic granules was recorded as m 1 The mass of particles that pass through the No. 1 sieve but not through the No. 5 sieve is recorded as m 2 , collect data to calculate the forming rate. (The sum of those that cannot pass through the No. 1 sieve and those that can pass through the No. 5 sieve shall not exceed 15%)

[0127]

[0128] 4.1.3 Solubility

[0129] According to the soluble granule test method in the 2020 edition of the Chinese Pharmacopoeia, 10 g of the sample was dissolved in hot water, stirred continuously, and a stopwatch was used to record the time required for the granules to be completely dissolved.

[0130] 4.1.4 Determination of moisture absorption

[0131] The moisture absorption rate is measured in a dryer at a relative humidity of 75% (supersaturated NaCl solution). Take an appropriate amount of sample and place it in a 30°C drying oven to maintain constant weight for 48 hours. Then spread the particles on a weighing bottle that has been weighed to a thickness of 3 mm. The mass of the weighing bottle is recorded as m0, and the total weight of the weighing bottle and particles is recorded as m 1 Place in a desiccator and weigh the mass after 48 hours, recorded as m 2 , substitute into the following formula to calculate.

[0132]

[0133] 4.1.5 Fluidity determination

[0134] Use the fixed funnel method, fix the funnel on the iron frame, and take another watch glass with the radius recorded as R, so that the funnel outlet and the center of the watch glass are on the same vertical line. Put an appropriate amount of particles into the funnel and let them fall naturally. When the cone of the falling particles coincides with the edge of the watch glass, record the height H of the cone and calculate the angle of repose.

[0135]

[0136] 4.1.6 Overall Rating

[0137] Forming rate, moisture absorption rate, fluidity and solubility are all important evaluation indicators of granule forming process, so each of them is given a weight of 25% for scoring

[0138]

[0139] C i represents the forming rate, C max indicates the maximum forming rate; X i represents the moisture absorption rate,X min represents the minimum moisture absorption rate; J i represents the angle of repose, J min represents the minimum angle of repose; R i represents solubility, R min indicates minimum solubility.

[0140] 4.2 Investigation of extract concentration density

[0141] According to the relative density determination method in the 2020 edition of the Chinese Pharmacopoeia, the specific gravity bottle method was used for determination. The decoction prepared by the optimal extraction process was concentrated to 150 ml, 125 ml, 100 ml, 75 ml, and 50 ml under reduced pressure at 60°C, and the relative density of the concentrate was determined. The results are shown in Table 11.

[0142] Table 11 Results of investigation on extract concentration density

[0143]

[0144] 4.3 Investigation of wetting agent concentration

[0145] Ethanol was selected as the wetting agent to prepare the particles, and the effect of ethanol solutions of different concentrations on particle formation was investigated. The results are shown in Table 12

[0146] Table 12 Wetting agent concentration investigation results

[0147]

[0148] According to the investigation results, 80% ethanol concentration showed good soft material properties, treatment conditions and particle uniformity. Therefore, about 80% ethanol should be selected as the wetting agent when preparing particles.

[0149] 4.4 Investigation of excipient types

[0150] Since this product is a hypoglycemic granule, sugar-free excipients should be selected when examining the types of excipients, such as β-cyclodextrin, mannitol, microcrystalline cellulose and sodium carboxymethyl cellulose. The excipients suitable for granulation are selected based on the molding rate, solubility, fluidity and moisture absorption rate, as shown in Table 13.

[0151] Table 13 Results of investigation on excipient types

[0152]

[0153] 4.5 Investigation of the amount of auxiliary materials

[0154] The excipient content in granules directly affects the molding and drug content. Too little excipient will affect the molding, while too much excipient will affect the drug content, resulting in an increased dosage. Choosing the right ratio of drug to excipient is crucial to the preparation of granules.

[0155] 4.6 Drying temperature investigation

[0156] Since the components of the sugar-reducing granules have good thermal stability, they are dried in a blast drying oven. After drying the wet granules at 50, 60, 65, 70, and 80 °C, it was found that the granules dried quickly but unevenly at 80 °C, and some granules were over-dried, charred, and blackened. The drying time was too long at 50-60 °C, and caking was likely to occur. The granules dried at around 65 °C were uniform and had a uniform color. The overall situation showed that the optimal drying temperature for the sugar-reducing granules was 65 °C.

[0157] 4.7 Determination of critical relative humidity

[0158] Evaluate the hygroscopicity and critical relative humidity of the granules prepared by the proposed process, and provide a test basis for the packaging and storage of the granules.

[0159] Take 14 portions of about 1g of granules, divide them into 7 groups, put them into 7 different relative humidity conditions, and store them in a constant temperature incubator at 25℃ for 108h. After taking them out, weigh them accurately and calculate the moisture absorption percentage.

[0160] According to the results of the investigation of critical relative humidity, an experimental basis can be provided for the environment of preparing particles. Take seven glass desiccators of the same size, add the substances listed in the following table, add a small amount of water to prepare a saturated salt solution, and place it in a drug stabilizer (temperature is 25 ℃, no relative humidity) for 24 h to obtain the corresponding constant temperature and humidity environment. Take about 2.0 g of hypoglycemic granules, spread them evenly on 24 weighing bottles with lids that have been weighed, accurately weigh the total weight, divide them into three groups, and place them in eight desiccators respectively. After being placed in a 25 ℃ constant temperature drug stabilizer for 7 days, the total weight of the granules and weighing bottles is measured, and the test results are calculated, see Table 14.

[0161] Table 14 Moisture absorption percentage of particles at different relative humidity

[0162]

[0163] 5. Determination of the preparation process of hypoglycemic granules

[0164] According to the prescription, eight medicinal materials, including black plum, kudzu root, astragalus, gardenia, yam, wolfberry, American ginseng and lotus seed core, were weighed in proportion, and boiled twice with water (10 times the amount of water for the first time and 8 times the amount of water for the second time), and each extraction was 75 minutes. Filter, combine the decoction, and concentrate under reduced pressure at 60°C to an extract with a relative density of 1.218~1.264. Add microcrystalline cellulose at a ratio of 3:2 of extract: auxiliary material, mix well, dry, crush, add appropriate amount of 80% ethanol as a wetting agent, wet granulate, dry at 65°C, arrange, and package to obtain the product.

[0165] 6 Dosage Design

[0166] After small-scale research, the daily dosage of the Jiangtang prescription is 60g. After extraction with the best water extraction process, the dry paste of a single prescription is 22.5g. After adding microcrystalline cellulose at a ratio of 3:2, it is adjusted to 37.5g. It is planned to be packaged into 12.5g per bag, and taken 3 times a day, 1 bag each time.

[0167] 7. Summary

[0168] The present invention determines the molding process. In the molding process of granules, a certain amount of auxiliary materials need to be added as fillers or lubricants in order to obtain better granule molding effects. In the process of studying the types of auxiliary materials, the present invention introduces molding rate, hygroscopicity, solubility, and angle of repose as corresponding evaluation indicators, and gives 25% of the indicator weight to score respectively, as the basis for the applicable or inapplicable types of auxiliary materials. Studies have shown that when microcrystalline cellulose is used as an auxiliary material, the comprehensive score is the highest. The optimal drug-excipient ratio is screened according to different proportions of pharmaceutical excipients, which is dry extract: excipient = 1:1. Under this drug-excipient ratio, the amount of particles finally prepared in each prescription is moderate, and both the quality of the particles themselves and the patient's compliance are taken into account. In addition, the present invention investigates the influence of the volume fraction of wetting agent ethanol and the drying temperature on particle molding, and it is found that when the ethanol ratio is 80%, the granulation is the best. When the drying temperature is 75°C, the color of the particles after drying is uniform, there is no scorching, and the drying efficiency is high, so it is determined as the optimal drying temperature. The critical relative humidity of the particles produced under this preparation process was also calculated, providing a certain theoretical basis for the ambient temperature of subsequent pilot production.

[0169] The beneficial effects of the present invention are demonstrated by the following efficacy tests.

[0170] Test Example 1 Blood sugar lowering test of the composition of the present invention

[0171] 1. Experimental Materials and Reagents

[0172]

[0173] 2. Experimental Methods

[0174] 2.1 Preparation of relevant solutions

[0175] Preparation of enzyme solution: Take appropriate amount of α-glucosidase and α-amylase in volumetric flasks, add PBS solution to dissolve and dilute to the scale to obtain α-glucosidase solution and α-amylase solution with mass concentration of 1 U / mL respectively.

[0176] Substrate preparation: Take an appropriate amount of p-nitrophenyl β-D-pyranogalactoside (PNPG) and place it in a volumetric flask, add PBS solution to dissolve and dilute to the scale to obtain a concentration of 5.0 mmol / LPNPG. Take an appropriate amount of soluble starch, add a certain amount of pure water to dissolve and boil until completely dissolved to obtain a 1% soluble starch solution.

[0177] Dosage:

[0178] Sugar-lowering granules (composition of the present invention): The raw materials of one prescription (60 g, 10 g of black plum, 10 g of astragalus, 10 g of kudzu root, 3 g of lotus seed core, 7 g of gardenia, 10 g of wolfberry, 7 g of yam, and 3 g of American ginseng) are subjected to the optimal preparation process to obtain 37.5 g of granules, which are taken three times a day, with each dose being 12.5 g, equivalent to 20 g of the raw materials.

[0179] (2) Plum-Astragalus: 20 g (Plum 10 g, Astragalus 10 g, equivalent to 4.6 times the dose of Plum-Astragalus in the composition of the present invention)

[0180] (3) Astragalus-American ginseng: 20 g (10 g of Astragalus and 10 g of American ginseng, equivalent to twice the dose of Astragalus-American ginseng in the composition of the present invention)

[0181] (4) Comparative patent group: Patent application number: CN201710704155.7 disclosed the formula except for the chemical drug chromium gluconate (American ginseng 35g; Astragalus 150g; Chinese yam 100g; Prunus mume 80g; Euryale ferox 70g; Hippophae rhamnoides 105g; Schisandra chinensis 55g; Ziziphus jujuba seed 35g; Pumpkin powder 100g; Pueraria root 75g; Licorice root 35g), the dosage was the same as that of group (1);

[0182] (5) Yam group

[0183] (6) Pueraria root alone group;

[0184] (7) Lotus seed core group;

[0185] The dosage of groups (5)-(7) is the same as that of group (1).

[0186] (Note: After the above formulas are extracted by heating and reflux, the water extracts are concentrated and diluted to ensure that the concentrations of each dose are consistent.)

[0187] 2.2 Determination of α-glucosidase inhibition rate

[0188] Acarbose was used as a positive control (the minimum dosage was 50 µL based on the positive control group acarbose). The specific operation was as follows: 50 µL of 1 U / mL α-glucosidase, 280 µL of 0.1 mol / L phosphate buffer (pH=6.8), 50 µL of acarbose, 50 µL of Wumeijiangtang granules water extract, 50 µL of Wumei-Huangqi herbal pair water extract, and 50 µL of American ginseng-Huangqi herbal pair water extract were taken, and the mixture was incubated at 37°C for 10 min. Then, 50 µL of 16 mmol / LPNPG was added, and the mixture was reacted at 37°C for 10 min. Finally, 0.2 mol / LNa 2 CO 3 200 μL of stop solution was added, and the absorbance was measured at 405 nm, which was recorded as the sample experimental group. Deionized water was used instead of the sample as the blank control group, and deionized water was used instead of the α-glucosidase solution as the sample control. The other steps were the same as above, and the sample blank and sample control were measured. All experiments were repeated three times. The α-glucosidase inhibition rate of samples with different concentrations was calculated according to formula (1).

[0189] Where: A 样品实验 , absorbance of sample solution, pNPG and α-glucosidase reaction solution; A 样品对照 The absorbance value of α-glucosidase was replaced by phosphate buffer; A 样品空白 , phosphate buffer was used instead of the absorbance of the sample solution. Each test was repeated 3 times, and the α-glucosidase inhibitory activity was calculated as [1-(A 样品试验 -A 样品对照 ) / A 样品空白 ]×100%.

[0190] 2.3 Determination of α-amylase inhibition rate

[0191] Acarbose was used as a positive control (the minimum dosage of 200 μL was determined based on the positive control group acarbose), and the α-amylase inhibition rate of the sample was determined by the DNS method. 200 μL of acarbose, Wumei Jiangtang granules water extract, Wumei-Huangqi herbal pair water extract, American ginseng-Huangqi herbal pair water extract and 100 μL of 1U / mL α-amylase solution were mixed, reacted at 37 °C for 20 min, 200 μL of 2.0% soluble starch was added, mixed, reacted at 37 °C for 10 min, 300 μL of DNS reagent was added, reacted in a boiling water bath for 5 min, 5 mL of distilled water was added for dilution, cooled, and the absorbance was measured at 540 nm, which was recorded as the sample experimental group. Deionized water was used instead of the sample as the blank control group, and deionized water was used instead of the α-amylase solution as the sample control. The other steps were the same as above, and the sample blank and sample control were measured. The α-amylase inhibition rate of samples with different concentrations was calculated according to formula (2).

[0192] Where: A 样品实验, absorbance of sample solution, 2.0% soluble starch and α-amylase reaction solution; A 样品对照 , phosphate buffer was used to replace the absorbance of α-amylase; A 样品空白 , phosphate buffer was used instead of the absorbance of the sample solution. Each test was repeated 3 times, and the α-amylase inhibitory activity was calculated as [1-(A 样品试验 -A 样品对照 ) / A 样品空白 ]×100%.

[0193] 3. Experimental results

[0194] 3.1 Results of α-glucosidase inhibitory activity assay

[0195] α-Glucosidase is one of the main glycoside hydrolases in the small intestine. It can quickly hydrolyze foods such as starch, thereby causing blood sugar to rise. Therefore, inhibiting the activity of α-glucosidase is an effective way to reduce hyperglycemia in diabetic patients. α-Glucosidase can react with colorless pNPG to generate yellow p-nitrophenol (PNP). PNP has a strong absorption at 405nm, and the sugar-reducing granules can inhibit the activity of α-glucosidase, reduce the generation of PNP, and make the yellow lighter, thereby reducing the absorbance value. Based on this, the inhibitory ability of the sugar-reducing granules on α-glucosidase can be judged. The test results are shown in Table 15.

[0196] Table 15 α-glucosidase inhibition rate

[0197] Serial number Acarbose Patent for this invention Comparative Patents Wumei-Astragalus American Ginseng-Astragalus Yam Pueraria root Lotus Seed Heart 1 89.57% 86.31% 76.83% 73.71% 72.63% 65.85% 69.51% 65.31% 2 90.51% 86.86% 76.42% 72.09% 71.95% 67.21% 69.92% 65.58% 3 89.97% 87.94% 77.78% 70.87% 70.05% 66.80% 68.16% 66.26% Mean 90.02% 87.04% <![CDATA[77.01% ** ]]> <![CDATA[72.22% ** ]]> <![CDATA[71.54% ** ]]> <![CDATA[66.62% ** ]]> <![CDATA[69.20% ** ]]> <![CDATA[65.72% ** ]]>

[0198] Note: Compared with the hypoglycemic granule group, * P <0.05, ** P <0.01.

[0199] 3.2 Results of α-amylase inhibitory activity determination

[0200] The sugar-reducing granules can inhibit the activity of α-amylase, weaken its ability to hydrolyze starch, and reduce the generation of reducing sugars, thereby reducing the orange-red substance generated by the reaction of reducing sugars with DNS and reducing the absorbance value. Based on this, the strength of the sugar-reducing granules' ability to inhibit α-amylase can be judged. The test results are shown in Table 16.

[0201] Table 16 α-amylase inhibition rate

[0202] Serial number Acarbose Patent for this invention Comparative Patents Wumei-Astragalus American Ginseng-Astragalus Yam Pueraria root Lotus Seed Heart 1 89.36% 85.43% 74.93% 67.37% 72.97% 65.97% 67.37% 65.83% 2 91.18% 88.10% 74.23% 71.29% 71.15% 66.39% 67.93% 64.57% 3 90.34% 86.13% 74.65% 70.03% 67.79% 67.37% 68.07% 64.85% Mean 90.29% 86.55% <![CDATA[74.60% * ]]> <![CDATA[69.56% ** ]]> <![CDATA[70.63% ** ]]> <![CDATA[66.57% ** ]]> <![CDATA[67.79% ** ]]> <![CDATA[65.08% ** ]]>

[0203] Note: Compared with the hypoglycemic granule group, * P <0.05, ** P<0.01.

[0204] Summary of experimental results:

[0205] The present invention mainly studies the preparation of hypoglycemic granules and proves the beneficial effects of the present invention through in vitro pharmacodynamic tests.

[0206] The preparation process of hypoglycemic granules includes extraction process, concentration process and molding process. Corresponding evaluation indicators are introduced in the three processes, and the AHP hierarchical analysis method is used to assign weights to the corresponding indicators. According to the comprehensive scoring results of each indicator component, the extraction process is determined to use 8 times the amount of water, extract twice, and each time for 75 minutes; the concentration process is determined to be concentrated under reduced pressure at 60°C; the molding process is to add microcrystalline cellulose in a ratio of 3:2 of extract: auxiliary materials, dry after mixing, crush, add an appropriate amount of 80% ethanol as a wetting agent, wet granulate, and dry at 65°C.

[0207] The results of in vitro pharmacodynamic tests showed that the inhibition rate of α-glucosidase inhibition activity of the composition of the present invention (Wumei Jiangtang Granules) reached 82.80%, which was significantly better than the single drug pair (Wumei-Astragalus: 66.49%, American ginseng-Astragalus: 65.42%) (*P<0.01). The inhibition rate of the composition was 81.57% of α-amylase inhibition activity, which was better than Wumei-Astragalus (72.04%) and American ginseng-Astragalus (72.47%) (*P<0.01), as well as the single raw material group and the reported patented formula, confirming the synergistic effect of multiple components.

Claims

1. A granule with hypoglycemic effect, characterized in that: The invention is prepared from the following raw materials in weight ratio: 8-12 parts of black plum, 8-12 parts of astragalus, 8-12 parts of kudzu root, 2.4-3.6 parts of lotus seed core, 5.6-8.4 parts of gardenia, 8-12 parts of wolfberry, 5.6-8.4 parts of yam and 2.4-3.6 parts of American ginseng. The preparation method of the granules comprises the following steps: a. weighing raw materials of various weight ratios; b. boiling with water twice, 10 times the amount of water for the first time and 8 times the amount of water for the second time, and extracting for 75 minutes each time; filtering, combining the decoctions, and concentrating under reduced pressure at 60°C to an extract with a relative density of 1.218 to 1.264, adding microcrystalline cellulose at a ratio of 3:2 of extract: auxiliary material, mixing well, drying, crushing, adding 80% ethanol as a wetting agent, wet granulating, drying at 65°C, sorting, and packaging to obtain the granules.

2. The granules with hypoglycemic effect according to claim 1, characterized in that: The invention is prepared from the following raw materials in the following weight ratio: 10 parts of black plum, 10 parts of astragalus, 10 parts of kudzu root, 3 parts of lotus seed core, 7 parts of gardenia, 10 parts of wolfberry, 7 parts of yam and 3 parts of American ginseng.

3. A method for preparing the granules with hypoglycemic effect according to claim 1 or 2, characterized in that: It includes the following steps: a. Weigh the raw materials of each weight ratio; b. Add water and boil twice, 10 times the amount of water for the first time and 8 times the amount of water for the second time, and extract for 75 minutes each time; filter, combine the decoctions, and concentrate under reduced pressure at 60°C to an extract with a relative density of 1.218 ~ 1.264, add microcrystalline cellulose at a ratio of 3:2 of extract: auxiliary material, mix well, dry, crush, add 80% ethanol as a wetting agent, wet granulate, dry at 65°C, arrange, and package to obtain.

4. Use of the granules according to claim 1 or 2 in the preparation of a medicament for treating diabetes.

5. The use according to claim 4, characterized in that: The medicine is a medicine for treating type II diabetes.

6. Use of the granules according to claim 1 or 2 in the preparation of health food that helps maintain healthy blood sugar levels.

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