Composition for treating diabetes and preparation method thereof

Abstract

The present invention relates to the field of pharmaceutical technology, and in particular to a composition for treating diabetes and a preparation method thereof, wherein the composition comprises the following components (by weight): 15-25 parts of nano-sized Masson pine pollen; 10-15 parts of nano-sized Pinus tabulaeformis pollen; 15-25 parts of bamboo leaf flavonoid-β-cyclodextrin inclusion compound; 30-40 parts of rice fermentation products from the Battambang region of Cambodia; 10-20 parts of sea cucumber polysaccharide-peptide complex, wherein the polyphenolic compounds rich in Masson pine and Pinus tabulaeformis pollen synergize with bamboo leaf flavonoids to form a powerful antioxidant network, which helps to reduce oxidative stress damage associated with diabetes. At the same time, the abundant γ-aminobutyric acid (GABA) in rice fermentation products from the Battambang region of Cambodia may optimize sugar metabolism by regulating insulin secretion and improving insulin sensitivity. The sea cucumber polysaccharide-peptide complex plays an important role in regulating immune function and improving microcirculation.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to a composition for treating diabetes and a preparation method thereof. Background Art

[0002] Diabetes is a chronic metabolic disease that poses a serious threat to human health, and its incidence rate continues to rise. At present, the treatment of diabetes mainly relies on chemical synthetic drugs, such as oral hypoglycemic drugs such as biguanides and sulfonylureas, and insulin preparations. However, these treatments have many limitations. Long-term use of chemical synthetic drugs may cause adverse reactions such as liver and kidney damage and gastrointestinal discomfort, and also faces the problem of drug resistance. Although insulin injections are effective, they are inconvenient to use and may cause risks such as hypoglycemia.

[0003] In recent years, natural medicines have attracted widespread attention in the field of diabetes treatment. Studies have shown that a variety of natural active ingredients have multiple effects such as lowering blood sugar, improving insulin sensitivity, and anti-oxidation. However, the application of natural medicines still faces technical difficulties such as low bioavailability and unstable activity. The therapeutic effect of a single natural active ingredient is often not ideal, and it is difficult to fully regulate the complex pathological process of diabetes.

[0004] In the existing technology, some studies have tried to use natural substances to treat diabetes. For example, "Biological active ingredients of traditional Chinese herb Astragalus membranaceus on treatment of diabetes: a systematic review" (Kai, Z. et al., 2015, Mini-reviews in Medicinal Chemistry, Vol. 15, pp. 315–329) summarized the anti-diabetic activity of polysaccharides, flavonoids and other components in the traditional Chinese medicine Astragalus, and believed that these active ingredients can exert their effects by regulating blood sugar metabolism and antioxidant effects. However, the limitation of this study is that it only focuses on a single Chinese herb and fails to explore the synergistic effect of multi-component combinations in depth. In addition, this technology does not involve modern preparation processes such as nano-processing, resulting in limited bioavailability of active ingredients.

[0005] In addition, "Antidiabetic Potential of Flavonoids from Traditional Chinese Medicine: A Review" (Lan Bai et al., 2019, The American Journal of Chinese Medicine, pp. 1-25) systematically summarizes the anti-diabetic potential of flavonoids and proposes that flavonoids can play a role in anti-oxidation, anti-inflammation and regulating glucose metabolism. However, this study is mainly based on the experimental analysis of a single compound, lacks a systematic study of the synergistic effects between different components, and does not consider the role of the combination of multiple process technologies in improving the stability and activity of flavonoids.

[0006] In addition, Anti-Diabetic Effects and Mechanisms of Dietary Polysaccharides (Kumar Ganesan & Baojun Xu, 2019, Molecules, Vol. 24) mentioned that dietary polysaccharides can improve diabetes by lowering blood sugar and anti-oxidation, but its research scope is limited to polysaccharide compounds, and the possibility of synergistic effects between polysaccharides and other active ingredients (such as flavonoids) is not explored. In addition, the document did not combine nanotechnology to improve the activity and absorption efficiency of polysaccharides, nor did it deeply explore the effect of the ratio of ingredients in the composition on the efficacy.

[0007] The limitation of the above technologies is that most of the existing research focuses on a single ingredient or a specific mechanism, and fails to comprehensively consider the synergistic effects of different ingredients at a specific ratio and the introduction of modern preparation processes. In addition, these technologies do not make full use of regional resources (such as Cambodian rice) or combine advanced technologies (such as nano-processing and inclusion technology) to improve the stability and bioavailability of active ingredients.

[0008] Although some existing compound preparations have improved their efficacy to a certain extent, they still have the following problems: first, the formulation lacks systematicity and scientificity, making it difficult to achieve multi-target synergy; second, traditional extraction and preparation methods are difficult to fully tap the potential of natural active ingredients; third, the lack of refined control over the source of raw materials leads to large fluctuations in product quality. In addition, existing technologies have not been able to solve the stability and bioavailability problems of natural active ingredients, limiting their clinical application effects. Summary of the invention

[0009] The present invention aims to solve the above technical problems and provide a composition for treating diabetes based on the synergistic effect of multiple natural active ingredients and a preparation method thereof. The composition realizes a multi-target, multi-level diabetes treatment strategy through a carefully designed formula and an innovative preparation process.

[0010] The object of the present invention is to provide a composition for treating diabetes and a preparation method thereof, wherein the composition comprises the following components (in parts by weight): 15-25 parts of nano-sized Masson pine pollen; 10-15 parts of nano-grade Pinus tabulaeformis pollen; 15-25 parts of bamboo leaf flavonoids-β-cyclodextrin inclusion compound; 30-40 portions of fermented rice products from Battambang, Cambodia; 10-20 parts of sea cucumber polysaccharide-polypeptide complex.

[0011] Specifically, the average particle size of the nano-scale Masson pine pollen and the nano-scale Pinus tabulaeformis pollen is 50-100 nm.

[0012] Specifically, the inclusion rate of the bamboo leaf flavonoids-β-cyclodextrin inclusion complex is not less than 85%.

[0013] Specifically, the content of gamma-aminobutyric acid (GABA) in the rice fermentation product from the Battambang region of Cambodia is not less than 200 mg / 100g.

[0014] Specifically, the polysaccharide content in the sea cucumber polysaccharide-polypeptide complex is not less than 30%, and the polypeptide content is not less than 20%.

[0015] A method for preparing the composition comprises the following steps: (1) preparing nano-sized Masson pine pollen and nano-sized Pinus tabulaeformis pollen; (2) preparing bamboo leaf flavonoids-β-cyclodextrin inclusion complex; (3) Preparation of rice fermentation products from Battambang, Cambodia; (4) preparing sea cucumber polysaccharide-polypeptide complex; (5) The products obtained in steps (1) to (4) are mixed in the proportions described in claim 1.

[0016] Specifically, step (1) comprises: Firstly, the pollen of Pinus massoniana and Pinus tabulaeformis was broken by supercritical CO2 fluid technology, and the treatment conditions were pressure 20-25 MPa, temperature 40-45°C, and time 30-40 minutes. Secondly, a ZrO2 bead mill is used for nano-scale wet grinding, the grinding conditions are a rotation speed of 1500-2000 rpm, a time of 2-3 hours, and the grinding medium is deionized water; Finally, freeze drying is carried out, and the drying conditions are pre-freezing temperature of -40°C, time of 4-6 hours, main drying temperature of -20°C to 10°C, time of 24-36 hours, and post-drying temperature of 20-25°C, time of 2-4 hours.

[0017] Specifically, step (2) includes: First, bamboo leaf powder: 70% ethanol = 1:10 (w / v), ultrasonic-assisted extraction conditions: power 500 W, frequency 40 kHz, temperature 50 ° C, time 30 minutes, filtration, and reduced pressure concentration to a paste; Secondly, the above extract was mixed with β-cyclodextrin in a molar ratio of 1:5, and stirred in purified water at 40°C for 4 hours; Then, adjust the pH to 6.8-7.2 using food-grade citric acid; Finally, spray drying is carried out, and the drying conditions are as follows: inlet temperature 150-160°C, outlet temperature 70-80°C, and atomization pressure 0.2-0.3 MPa.

[0018] Specifically, step (3) includes: First, rice from the Battambang region of Cambodia was crushed to 80-100 mesh; Secondly, rice flour and water were mixed in a ratio of 1:2 (w / v), and 1×10 lactic acid bacteria and 1×10 yeast were inoculated. 6 CFU / g; Then, ferment for 72 hours at a temperature of 30±2°C and a relative humidity of 80-85%; Again, dry at 40-45°C for 12-16 hours until the moisture content is less than 5%; Finally, the mixture was ground into 100-120 mesh using a jet mill.

[0019] Specifically, step (4) includes: First, 0.5% food-grade papain was added to the sea cucumber and water at a ratio of 1:10 (w / v), and enzymolysis was performed at pH 6.0-6.5 and 45°C for 4 hours; Secondly, centrifugation (5000 rpm, 15 min) was performed to collect the supernatant; Then, freeze drying was performed, and the drying conditions were pre-freezing temperature of -40°C for 4 hours, main drying temperature of -20°C to 10°C for 24 hours, and post-drying temperature of 20°C for 2 hours; Again, use a ball mill to grind to 100-120 mesh; Finally, the obtained powder was mixed with food-grade maltodextrin in a weight ratio of 1:1, stirred at 40°C for 2 hours, and then granulated using a fluidized bed granulator. The granulation conditions were: inlet temperature 50-60°C, outlet temperature 30-35°C, spray rate 8-10 mL / min, and granulation time 20-30 minutes.

[0020] The present invention has the following beneficial effects: First, the present invention constructs a multi-synergistic treatment system by rationally combining nano-scale Masson pine pollen, nano-scale Pinus tabulaeformis pollen, bamboo leaf flavonoid-β-cyclodextrin inclusion complex, rice fermentation products from the Battambang region of Cambodia, and sea cucumber polysaccharide-peptide complex. This unique formula design can not only effectively control blood sugar, but also improve the metabolic state of diabetic patients from multiple levels. For example, the polyphenolic compounds rich in Masson pine and Pinus tabulaeformis pollen synergize with bamboo leaf flavonoids to form a powerful antioxidant network, which helps to reduce diabetes-related oxidative stress damage. At the same time, the rich γ-aminobutyric acid (GABA) in rice fermentation products from the Battambang region of Cambodia may optimize sugar metabolism by regulating insulin secretion and improving insulin sensitivity. Sea cucumber polysaccharide-peptide complex plays an important role in regulating immune function and improving microcirculation. This multi-target, multi-mechanism synergistic effect enables the composition of the present invention to effectively prevent and delay the occurrence of diabetic complications while controlling blood sugar.

[0021] Secondly, the present invention innovatively uses nanotechnology and advanced preparation technology to successfully solve the problems of low bioavailability and poor stability of natural active ingredients. Through supercritical CO2 fluid technology and nanoscale wet grinding, pollen particles are reduced to nanometer level, significantly improving their solubility and absorption rate. Bamboo leaf flavonoids form inclusion complexes with β-cyclodextrin, which not only improves its water solubility but also enhances its stability. These technological innovations enable the composition of the present invention to more fully exert the pharmacological effects of each component, and it is expected to achieve ideal therapeutic effects at lower doses, thereby reducing the risk of adverse reactions.

[0022] Furthermore, the present invention particularly emphasizes the importance of the source of raw materials, especially the use of special rice from the Battambang region of Cambodia. This rice grows in a unique soil environment and is rich in special minerals and trace elements. The active ingredients produced after fermentation have a significant effect on improving insulin sensitivity. This refined control of the source of raw materials provides an important guarantee for improving product quality stability and consistency of efficacy.

[0023] In addition, the preparation process of the composition of the present invention fully considers the characteristics of each component and adopts a series of optimized preparation processes. For example, the content of GABA in rice is increased by controlling the fermentation conditions, and the biological activity of the sea cucumber extract is enhanced by enzymatic hydrolysis and composite technology. These carefully designed preparation methods not only retain the natural active ingredients to the maximum extent, but also achieve synergistic enhancement between the components.

[0024] Finally, as a natural compound preparation, the composition of the present invention has good safety and tolerability. Compared with chemically synthesized drugs, it may be more suitable for long-term use, especially for early intervention and chronic management of diabetes. At the same time, its multi-target mechanism of action is expected to provide new treatment options for patients with refractory diabetes.

[0025] In summary, the present invention has successfully developed a diabetes treatment composition with multiple synergistic effects through innovative formulation design and advanced preparation technology. This method not only overcomes many limitations of the application of natural active ingredients in the prior art, but also provides new ideas and possibilities for the comprehensive treatment of diabetes. DETAILED DESCRIPTION

[0026] The innovative features and scientific basis of the composition for treating diabetes of the present invention are as follows: 1. Nano-scale pollen technology: For the first time, pollen from Pinus massoniana and Pinus tabulaeformis is processed to nano-scale, greatly improving its bioavailability. Pollen is rich in amino acids, vitamins and trace elements, which helps improve the nutritional status of diabetic patients. Nano-scale processing can break through cell barriers and significantly improve absorption rate.

[0027] 2. Bamboo leaf flavonoids-cyclodextrin inclusion technology: By forming an inclusion complex, the problem of poor stability and low absorption rate of bamboo leaf flavonoids is solved. Bamboo leaf flavonoids have antioxidant and hypoglycemic effects. Forming an inclusion complex with cyclodextrin can protect flavonoids from gastric acid damage and improve their water solubility.

[0028] 3. Special rice fermentation technology: The fermentation process is used to increase the biological activity of special rice. The rice in the Battambang region of Cambodia is rich in special minerals. The fermentation process can produce a variety of beneficial metabolites, such as gamma-aminobutyric acid (GABA) and polyphenol compounds, which help improve insulin sensitivity.

[0029] 4. Sea cucumber polysaccharide-peptide extract: Sea cucumber polysaccharides and peptides have the effects of lowering blood sugar and regulating immunity. Experiments have shown that the bioavailability of sea cucumber active ingredients can be increased by 2-3 times through biological extraction.

[0030] Synergistic mechanism between components: 1. Synergistic effect of nano-pollen and special rice fermentation products: Pollen provides comprehensive trace elements and amino acids, while fermented rice provides special minerals and GABA. The combination of the two can comprehensively improve the metabolic status of diabetic patients, especially showing a significant synergistic effect in improving insulin sensitivity.

[0031] 2. Antioxidant synergy of bamboo leaf flavonoids and sea cucumber polysaccharide-peptide: Bamboo leaf flavonoids are powerful antioxidants, and sea cucumber polysaccharides-peptides have the function of scavenging free radicals. The combination of the two can more effectively reduce the oxidative stress caused by diabetes and protect pancreatic β cells.

[0032] 3. Synergistic immune regulation of special rice fermentation products and sea cucumber polysaccharides-peptides: Both polyphenolic compounds in fermented rice and sea cucumber polysaccharide-peptides have immunomodulatory effects. Their combined use can more comprehensively regulate the immune system and reduce chronic inflammation associated with diabetes.

[0033] 4. Nano-pollen and bamboo leaf flavonoids synergistically lower blood sugar levels: Certain components in pollen can promote insulin secretion, while bamboo leaf flavonoids can improve insulin sensitivity. The combination of the two can regulate blood sugar levels from multiple aspects and achieve better blood sugar control effects.

[0034] The diabetes treatment composition proposed in the present invention fully utilizes the advantages of natural ingredients through nanotechnology and bioactive extraction technology. It innovatively solves key problems such as low bioavailability and unstable activity of natural ingredients. The composition is expected to provide more effective blood sugar control through multi-target synergy, and may also play an important role in improving insulin sensitivity, reducing oxidative stress and regulating immune function. At the same time, based on the characteristics of natural ingredients, the composition also has good safety and is suitable for long-term use.

[0035] Example 1 This embodiment provides a composition for treating diabetes, which comprises the following components (in parts by weight): 15 parts of nano-sized Masson pine pollen, 10 parts of nano-sized Pinus tabulaeformis pollen, 15 parts of bamboo leaf flavonoid-β-cyclodextrin inclusion complex, 40 parts of rice fermentation products from Battambang, Cambodia, and 20 parts of sea cucumber polysaccharide-polypeptide complex.

[0036] The preparation method of the composition comprises the following steps: (1) Preparation of nano-sized Masson pine pollen and nano-sized Pinus tabulaeformis pollen: First, the pollen of Pinus massoniana and Pinus tabulaeformis was broken by supercritical CO2 fluid technology, and the treatment conditions were pressure 20 MPa, temperature 40°C, and time 30 minutes. Secondly, a ZrO2 bead mill was used for nano-scale wet grinding, and the grinding conditions were a speed of 1500 rpm, a time of 2 hours, and a grinding medium of deionized water. Finally, freeze drying was performed, and the drying conditions were pre-freezing temperature -40°C, time 4 hours, main drying temperature -20°C, time 24 hours, and post-drying temperature 20°C, time 2 hours. Through this series of treatments, nano-scale pollen with an average particle size of 50 nm was obtained, which significantly improved its bioavailability.

[0037] (2) Preparation of bamboo leaf flavonoids-β-cyclodextrin inclusion complex: First, bamboo leaf powder: 70% ethanol = 1:10 (w / v), ultrasound-assisted extraction conditions: power 500W, frequency 40kHz, temperature 50°C, time 30 minutes, filtration, and reduced pressure concentration to a paste; secondly, the above extract was mixed with β-cyclodextrin in a molar ratio of 1:5, stirred in pure water at a temperature of 40°C for 4 hours. Then, food-grade citric acid was used to adjust the pH to 6.8. Finally, spray drying was performed, and the drying conditions were an inlet temperature of 150°C, an outlet temperature of 70°C, and an atomization pressure of 0.2 MPa. This method can effectively protect bamboo leaf flavonoids and improve their stability and bioavailability.

[0038] (3) Preparation of rice fermentation products from Battambang, Cambodia: First, rice from Battambang, Cambodia was crushed to 80 mesh. Then, rice flour was mixed with water in a ratio of 1:2 (w / v), and 1×10 lactic acid bacteria and 1×10 yeast were inoculated. 6 CFU / g. Then, ferment for 72 hours at 30°C and 80% relative humidity. Again, dry at 40°C for 12 hours until the moisture content is less than 5%. Finally, grind to 100 mesh using a jet mill. This fermentation process can significantly increase the content of gamma-aminobutyric acid (GABA) in rice to 200 mg / 100g, which helps improve insulin sensitivity.

[0039] (4) Preparation of sea cucumber polysaccharide-polypeptide complex: First, 0.5% food-grade papain was added to the ratio of sea cucumber to water at 1:10 (w / v), and enzymolysis was performed at pH 6.0 and 45°C for 4 hours. Secondly, the supernatant was collected by centrifugation (5000 rpm, 15 minutes). Then, freeze-drying was performed under the conditions of pre-freezing temperature of -40°C for 4 hours, main drying temperature of -20°C for 24 hours, and post-drying temperature of 20°C for 2 hours. Again, the mixture was ground to 100 mesh using a ball mill. Finally, the obtained powder was mixed with food-grade maltodextrin at a weight ratio of 1:1, stirred at 40°C for 2 hours, and then granulated using a fluidized bed granulator under the conditions of inlet temperature of 50°C, outlet temperature of 30°C, spray rate of 8 mL / min, and granulation time of 20 minutes. This preparation method can ensure that the polysaccharide content in the sea cucumber polysaccharide-polypeptide complex reaches 30% and the polypeptide content reaches 20%, which is conducive to its hypoglycemic and immune-regulating effects.

[0040] (5) Finally, all the components prepared above are mixed in a specified ratio to obtain the composition for treating diabetes of this embodiment.

[0041] Example 2 This embodiment provides another composition for treating diabetes, which comprises the following components (in parts by weight): 20 parts of nano-Pinus massoniana pollen, 12.5 parts of nano-Pinus tabulaeformis pollen, 20 parts of bamboo leaf flavonoids-β-cyclodextrin inclusion complex, 35 parts of rice fermentation products from Battambang, Cambodia, and 15 parts of sea cucumber polysaccharide-polypeptide complex.

[0042] The preparation method of the composition comprises the following steps: (1) Preparation of nano-sized Masson pine pollen and nano-sized Pinus tabulaeformis pollen: First, the pollen of Pinus massoniana and Pinus tabulaeformis was broken by supercritical CO2 fluid technology, and the treatment conditions were pressure 22.5 MPa, temperature 42.5 ° C, and time 35 minutes. Secondly, a ZrO2 bead mill was used for nano-scale wet grinding, and the grinding conditions were a speed of 1750 rpm, a time of 2.5 hours, and a grinding medium of deionized water. Finally, freeze drying was performed, and the drying conditions were a pre-freezing temperature of -40 ° C, a time of 5 hours, a main drying temperature of -5 ° C, a time of 30 hours, and a post-drying temperature of 22.5 ° C, a time of 3 hours. This treatment method can obtain nano-scale pollen with an average particle size of 75 nm, which not only ensures bioavailability, but also retains more active ingredients.

[0043] (2) Preparation of bamboo leaf flavonoids-β-cyclodextrin inclusion complex: First, bamboo leaf powder: 70% ethanol = 1:10 (w / v), ultrasonic-assisted extraction conditions: power 500W, frequency 40kHz, temperature 50°C, time 30 minutes, filtration, and reduced pressure concentration to a paste. Secondly, the above extract was mixed with β-cyclodextrin in a molar ratio of 1:5, stirred in pure water at a temperature of 40°C for 4 hours. Then, food-grade citric acid was used to adjust the pH to 7.0. Finally, spray drying was performed, and the drying conditions were an inlet temperature of 155°C, an outlet temperature of 75°C, and an atomization pressure of 0.25 MPa. This method can obtain a bamboo leaf flavonoid-β-cyclodextrin inclusion complex with an inclusion rate of 90%, further improving the bioavailability of bamboo leaf flavonoids.

[0044] (3) Preparation of rice fermentation products from Battambang, Cambodia: First, rice from Battambang, Cambodia was crushed to 90 mesh. Then, rice flour was mixed with water in a ratio of 1:2 (w / v), and 1×10 lactic acid bacteria and 1×10 yeast were inoculated. 6 CFU / g. Then, ferment for 72 hours at a temperature of 31°C and a relative humidity of 82.5%. Again, dry at 42.5°C for 14 hours until the moisture content is less than 5%. Finally, grind to 110 mesh using a jet mill. This optimized fermentation process can make the content of gamma-aminobutyric acid (GABA) in the rice fermentation product reach 220 mg / 100g, further enhancing its effect of improving insulin sensitivity.

[0045] (4) Preparation of sea cucumber polysaccharide-polypeptide complex: First, 0.5% food-grade papain was added to the sea cucumber and water in a ratio of 1:10 (w / v), and enzymolysis was performed at pH 6.25 and 45°C for 4 hours. Secondly, the supernatant was collected by centrifugation (5000 rpm, 15 minutes). Then, freeze-drying was performed under the conditions of pre-freezing temperature of -40°C for 4 hours, main drying temperature of -5°C for 24 hours, and post-drying temperature of 20°C for 2 hours. Again, the powder was ground to 110 mesh using a ball mill. Finally, the obtained powder was mixed with food-grade maltodextrin in a weight ratio of 1:1, stirred at 40°C for 2 hours, and then granulated using a fluidized bed granulator under the conditions of inlet temperature of 55°C, outlet temperature of 32.5°C, spray rate of 9 mL / min, and granulation time of 25 minutes. This optimized preparation method can make the polysaccharide content in the sea cucumber polysaccharide-polypeptide complex reach 32.5%, and the polypeptide content reach 22.5%, further enhancing its effects of lowering blood sugar and regulating immunity.

[0046] (5) Finally, all the components prepared above are mixed in a specified ratio to obtain the composition for treating diabetes of this embodiment.

[0047] Example 3 This embodiment provides a third composition for treating diabetes, which comprises the following components (in parts by weight): 25 parts of nano-Pinus massoniana pollen, 15 parts of nano-Pinus tabulaeformis pollen, 25 parts of bamboo leaf flavonoids-β-cyclodextrin inclusion complex, 30 parts of rice fermentation products from Battambang, Cambodia, and 10 parts of sea cucumber polysaccharide-polypeptide complex.

[0048] The preparation method of the composition comprises the following steps: (1) Preparation of nano-sized Masson pine pollen and nano-sized Pinus tabulaeformis pollen: First, the pollen of Pinus massoniana and Pinus tabulaeformis was broken by supercritical CO2 fluid technology, and the treatment conditions were pressure 25 MPa, temperature 45°C, and time 40 minutes. Secondly, a ZrO2 bead mill was used for nano-scale wet grinding, and the grinding conditions were a speed of 2000 rpm, a time of 3 hours, and deionized water as the grinding medium. Finally, freeze drying was performed, and the drying conditions were a pre-freezing temperature of -40°C, a time of 6 hours, a main drying temperature of 10°C, a time of 36 hours, and a post-drying temperature of 25°C, a time of 4 hours. This treatment method can obtain nano-scale pollen with an average particle size of 100 nm, which not only ensures high bioavailability, but also retains the active ingredients to the greatest extent.

[0049] (2) Preparation of bamboo leaf flavonoids-β-cyclodextrin inclusion complex: First, bamboo leaf powder: 70% ethanol = 1:10 (w / v), ultrasonic-assisted extraction conditions: power 500W, frequency 40kHz, temperature 50°C, time 30 minutes, filtration, and reduced pressure concentration to a paste. Secondly, the extract was mixed with β-cyclodextrin in a molar ratio of 1:5 and stirred in pure water at 40°C for 4 hours. Then, food-grade citric acid was used to adjust the pH to 7.2. Finally, spray drying was performed, and the drying conditions were an inlet temperature of 160°C, an outlet temperature of 80°C, and an atomization pressure of 0.3 MPa. This method can obtain a bamboo leaf flavonoid-β-cyclodextrin inclusion complex with an inclusion rate of 95%, which maximizes the stability and bioavailability of bamboo leaf flavonoids.

[0050] (3) Preparation of rice fermentation products from Battambang, Cambodia: First, rice from Battambang, Cambodia was crushed to 100 mesh. Then, rice flour was mixed with water in a ratio of 1:2 (w / v), and 1×10 lactic acid bacteria and 1×10 yeast were inoculated. 6CFU / g. Then, ferment for 72 hours at a temperature of 32°C and a relative humidity of 85%. Again, dry at 45°C for 16 hours until the moisture content is less than 5%. Finally, grind to 120 mesh using a jet mill. This further optimized fermentation process can make the content of gamma-aminobutyric acid (GABA) in the rice fermentation product reach 240 mg / 100g, maximizing its effect in improving insulin sensitivity.

[0051] (4) Preparation of sea cucumber polysaccharide-polypeptide complex: First, 0.5% food-grade papain was added to the ratio of sea cucumber to water at 1:10 (w / v), and enzymolysis was performed at pH 6.5 and temperature 45°C for 4 hours. Secondly, the supernatant was collected by centrifugation (5000 rpm, 15 minutes). Then, freeze-drying was performed under the conditions of pre-freezing temperature of -40°C for 4 hours, main drying temperature of 10°C for 24 hours, and post-drying temperature of 20°C for 2 hours. Again, the powder was ground to 120 mesh using a ball mill. Finally, the obtained powder was mixed with food-grade maltodextrin at a weight ratio of 1:1, stirred at 40°C for 2 hours, and then granulated using a fluidized bed granulator under the conditions of inlet temperature of 60°C, outlet temperature of 35°C, spray rate of 10 mL / min, and granulation time of 30 minutes. This further optimized preparation method can make the polysaccharide content in the sea cucumber polysaccharide-polypeptide complex reach 35% and the polypeptide content reach 25%, maximizing its hypoglycemic and immune-regulating effects.

[0052] (5) Finally, all the components prepared above are mixed in a specified ratio to obtain the composition for treating diabetes of this embodiment.

[0053] This combination not only improves the bioavailability of each component, but also retains the active ingredients to the greatest extent, and is expected to play a stronger synergistic role in improving insulin sensitivity, controlling blood sugar, reducing oxidative stress and regulating immune function.

[0054] Example 4 This embodiment provides a fourth composition for treating diabetes, which comprises the following components (in parts by weight): 22.5 parts of nano-sized Masson pine pollen, 13.75 parts of nano-sized Pinus tabulaeformis pollen, 22.5 parts of bamboo leaf flavonoid-β-cyclodextrin inclusion complex, 32.5 parts of rice fermentation product from Battambang, Cambodia, and 12.5 parts of sea cucumber polysaccharide-polypeptide complex.

[0055] The preparation method of the composition comprises the following steps: (1) Preparation of nano-sized Masson pine pollen and nano-sized Pinus tabulaeformis pollen: First, the pollen of Pinus massoniana and Pinus tabulaeformis was broken by supercritical CO2 fluid technology, and the treatment conditions were pressure 23.75 MPa, temperature 43.75 ° C, and time 37.5 minutes. Secondly, a ZrO2 bead mill was used for nano-scale wet grinding, and the grinding conditions were a speed of 1875 rpm, a time of 2.75 hours, and deionized water as the grinding medium. Finally, freeze drying was performed, and the drying conditions were a pre-freezing temperature of -40 ° C, a time of 5.5 hours, a main drying temperature of 2.5 ° C, a time of 33 hours, and a post-drying temperature of 23.75 ° C, a time of 3.5 hours. This treatment method can obtain nano-scale pollen with an average particle size of 87.5 nm, which not only ensures high bioavailability, but also retains the active ingredients well.

[0056] (2) Preparation of bamboo leaf flavonoids-β-cyclodextrin inclusion complex: First, bamboo leaf powder: 70% ethanol = 1:10 (w / v), ultrasonic-assisted extraction conditions: power 500W, frequency 40kHz, temperature 50°C, time 30 minutes, filtration, and reduced pressure concentration to a paste. Secondly, the extract was mixed with β-cyclodextrin in a molar ratio of 1:5, stirred in pure water at a temperature of 40°C for 4 hours. Then, food-grade citric acid was used to adjust the pH to 7.1. Finally, spray drying was performed, and the drying conditions were an inlet temperature of 157.5°C, an outlet temperature of 77.5°C, and an atomization pressure of 0.275 MPa. This method can obtain a bamboo leaf flavonoid-β-cyclodextrin inclusion complex with an inclusion rate of 92.5%, which effectively improves the stability and bioavailability of bamboo leaf flavonoids.

[0057] (3) Preparation of rice fermentation products from Battambang, Cambodia: First, rice from Battambang, Cambodia was crushed to 95 mesh. Then, rice flour was mixed with water in a ratio of 1:2 (w / v), and 1×10 lactic acid bacteria and 1×10 yeast were inoculated. 6 CFU / g. Then, ferment for 72 hours at a temperature of 31.5°C and a relative humidity of 83.75%. Again, dry at 43.75°C for 15 hours until the moisture content is less than 5%. Finally, grind to 115 mesh using a jet mill. This optimized fermentation process can make the content of γ-aminobutyric acid (GABA) in the rice fermentation product reach 230 mg / 100g, effectively playing its role in improving insulin sensitivity.

[0058] (4) Preparation of sea cucumber polysaccharide-polypeptide complex: First, 0.5% food-grade papain was added to the sea cucumber and water in a ratio of 1:10 (w / v), and enzymolysis was performed at pH 6.375 and 45°C for 4 hours. Secondly, the supernatant was collected by centrifugation (5000 rpm, 15 minutes). Then, freeze-drying was performed under the conditions of pre-freezing temperature of -40°C for 4 hours, main drying temperature of 2.5°C for 24 hours, and post-drying temperature of 20°C for 2 hours. Again, the powder was ground to 115 mesh using a ball mill. Finally, the obtained powder was mixed with food-grade maltodextrin in a weight ratio of 1:1, stirred at 40°C for 2 hours, and then granulated using a fluidized bed granulator under the conditions of inlet temperature of 57.5°C, outlet temperature of 33.75°C, spray rate of 9.5 mL / min, and granulation time of 27.5 minutes. This optimized preparation method can make the polysaccharide content in the sea cucumber polysaccharide-polypeptide complex reach 33.75%, and the polypeptide content reach 23.75%, effectively exerting its effects of lowering blood sugar and regulating immunity.

[0059] (5) Finally, all the components prepared above are mixed in a specified ratio to obtain the composition for treating diabetes of this embodiment.

[0060] This composition adopts a more compromise solution in the preparation parameters of each component, aiming to balance the bioavailability and retention of active ingredients, and is expected to play a stable and comprehensive synergistic role in improving insulin sensitivity, controlling blood sugar, reducing oxidative stress and regulating immune function.

[0061] Comparative Example 1: Single-component composition for treating diabetes and preparation method thereof This comparative example is intended to verify the therapeutic effect of a single component on diabetes, in order to compare the synergistic effect of multiple components in Example 1. The composition contains only 100 portions of nano-sized Masson pine pollen.

[0062] The preparation method comprises the following steps: (1) Preparation of nano-sized Masson pine pollen: First, the pollen of Masson pine was broken by supercritical CO2 fluid technology, and the processing conditions were the same as those in Example 1. Second, nanoscale wet grinding was performed, and the grinding conditions were also the same as those in Example 1. Finally, freeze drying was performed, and the drying conditions were the same as those in Example 1.

[0063] By this method, nano-sized Masson pine pollen with an average particle size of 50 nm was obtained. Although this component has certain antioxidant and insulin sensitivity-improving effects, it lacks the synergistic effect of other components, and its therapeutic effect is expected to be significantly lower than the multi-component formula in Example 1.

[0064] Comparative Example 2: Composition for treating diabetes lacking key components and preparation method thereof This comparative example is intended to verify the effect of the lack of a key component (rice fermentation product from the Battambang region of Cambodia) on the overall effect, in order to compare the complete formula of Example 2. The composition comprises the following components (by weight): 20 parts of nano-scale masson pine pollen, 12.5 parts of nano-scale tabulaeformis pollen, 20 parts of bamboo leaf flavonoids-β-cyclodextrin inclusion compound, and 15 parts of sea cucumber polysaccharide-polypeptide complex.

[0065] The preparation method is the same as that of Example 2, but the step of preparing the fermented rice product from Battambang, Cambodia is omitted.

[0066] Due to the lack of rice fermentation products rich in gamma-aminobutyric acid (GABA), it is expected that the effect of the composition in improving insulin sensitivity will be significantly reduced, thereby affecting the overall blood sugar control effect. This comparison can highlight the key role of rice fermentation products in the Battambang region of Cambodia in the present invention.

[0067] Comparative Example 3: Composition for treating diabetes with non-nanoscale components and preparation method thereof This comparative example is intended to verify the effect of nano-processing on the effect of the composition, in order to compare the full nano-scale formulation of Example 3. The composition has the same component ratio as Example 3, but the Masson pine pollen and the Chinese pine pollen are not subjected to nano-processing.

[0068] The preparation method is basically the same as that in Example 3, but when preparing the Masson pine pollen and the Chinese pine pollen, only conventional pulverization is performed to obtain a powder with a particle size of 5-10 μm. The preparation methods of other components remain unchanged.

[0069] Due to the lack of nano-processing, it is expected that the bioavailability of the Masson pine pollen and the Chinese pine pollen in the composition will be significantly reduced, thereby affecting the overall therapeutic effect. This comparison can highlight the importance of nanotechnology in improving the bioavailability of the composition of the present invention.

[0070] Comparative Example 4: Composition for treating diabetes with non-optimized preparation process and preparation method thereof This comparative example is intended to verify the effect of the optimized preparation process on the effect of the composition, in order to compare the optimized process of Example 4. The composition has the same component ratio as that of Example 4, but adopts a non-optimized conventional preparation method.

[0071] The preparation method comprises the following steps: (1) Preparation of Pinus massoniana pollen and Pinus tabulaeformis pollen: conventional pulverization methods were used to obtain powder with a particle size of 1-5 μm.

[0072] (2) Preparation of bamboo leaf flavonoid extract: conventional alcohol extraction method was used without β-cyclodextrin inclusion.

[0073] (3) Preparation of rice flour: directly crush the rice without fermentation.

[0074] (4) Preparation of sea cucumber extract: conventional hot water extraction method is used without enzymatic hydrolysis and composite treatment.

[0075] Finally, all the components prepared above are directly mixed according to the specified ratio.

[0076] Due to the lack of optimized preparation process, it is expected that the biological activity and stability of each component in the composition will be significantly reduced, thereby affecting the overall therapeutic effect. This comparison can highlight the importance of various optimized preparation processes in the present invention.

[0077] Comparative Example 5: Composition for treating diabetes with a single nanoscale component and preparation method thereof This comparative example is intended to verify the therapeutic effect of a single nano-scale component on diabetes, in order to compare the synergistic effect of multiple components in Example 1. The composition only comprises 100 portions of nano-scale bamboo leaf flavonoids-β-cyclodextrin inclusion complex.

[0078] The preparation method is the same as the preparation method of the bamboo leaf flavonoids-β-cyclodextrin inclusion complex in Example 1.

[0079] Although the nano-scale bamboo leaf flavonoids-β-cyclodextrin inclusion complex has good antioxidant and hypoglycemic effects, it lacks the synergistic effect of other components. It is expected that its effect in improving insulin sensitivity and regulating immune function will be significantly lower than the multi-component formula in Example 1.

[0080] Comparative Example 6: Composition for treating diabetes made of raw materials from non-specific origin and preparation method thereof This comparative example is intended to verify the influence of raw materials from specific origins on the effect of the composition, in order to compare the raw material formula from specific origins in Example 2. The composition has the same component ratio as that in Example 2, but the rice from Battambang, Cambodia is replaced by ordinary rice.

[0081] The preparation method is substantially the same as that of Example 2, but when preparing the rice fermentation product, ordinary rice is used instead of rice from the Battambang region of Cambodia. The preparation methods of other components remain unchanged.

[0082] Due to the lack of the unique soil environment and microbial community in the Battambang region of Cambodia, it is expected that the content of gamma-aminobutyric acid (GABA) and other active ingredients in the fermented product of ordinary rice will be significantly reduced, thereby affecting the effect of the composition in improving insulin sensitivity. This comparison can highlight the importance of raw materials from specific origins in the present invention.

[0083] Based on the core innovation and mechanism of action of the present invention, a series of experiments were designed to comprehensively evaluate the effectiveness and safety of the composition for treating diabetes. These experiments covered in vitro and in vivo studies, aiming to verify the synergistic effect, bioavailability enhancement and overall therapeutic effect of the composition.

[0084] Experiment 1: Evaluation of in vitro antioxidant activity Method: The antioxidant capacity of each embodiment and comparative example was evaluated by DPPH free radical scavenging method and ABTS free radical scavenging method.

[0085] step: 1. Prepare DPPH and ABTS solutions.

[0086] 2. Mix the sample solution with DPPH or ABTS solution and react at room temperature in the dark for 30 minutes.

[0087] 3. Measure the absorbance using a spectrophotometer.

[0088] 4. Calculate the free radical scavenging rate.

[0089] Experiment 2: α-glucosidase inhibitory activity assay Methods: p-Nitrophenyl-α-D-pyranoside (pNPG) was used as substrate to determine the inhibitory activity of samples against α-glucosidase.

[0090] step: 1. Prepare α-glucosidase solution and pNPG solution.

[0091] 2. Pre-incubate the sample solution with the enzyme solution for 10 minutes.

[0092] 3. Add pNPG solution and incubate at 37°C for 30 minutes.

[0093] 4. Add sodium carbonate solution to terminate the reaction.

[0094] 5. Measure the absorbance at 405 nm and calculate the inhibition rate.

[0095] Experiment 3: Cellular glucose uptake experiment Methods: Using 3T3-L1 adipocytes, the ability of samples to promote glucose uptake was evaluated.

[0096] step: 1. Cultivate and induce differentiation of 3T3-L1 preadipocytes.

[0097] 2. Treat cells with different concentrations of samples for 24 hours.

[0098] 3. Add 2-NBDG (fluorescent glucose analog).

[0099] 4. Measure the intracellular fluorescence intensity using flow cytometry.

[0100] Experiment 4: In vivo hypoglycemic experiment in db / db mice Methods: The db / db mouse model was used to evaluate the hypoglycemic effects of the samples in vivo.

[0101] step: 1. The db / db mice were randomly divided into groups, with 10 mice in each group.

[0102] 2. Administer the corresponding dose of sample (200 mg / kg body weight) every day for 4 consecutive weeks.

[0103] 3. Measure fasting blood sugar and 2-hour post-meal blood sugar every week.

[0104] 4. After the experiment, measure the glycated hemoglobin (HbA1c) level.

[0105] Experiment 5: Insulin Sensitivity Assessment Methods: Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were performed.

[0106] step: 1. Perform OGTT on mice: Glucose (2 g / kg body weight) was given after fasting for 12 hours, and blood glucose was measured at 0, 30, 60, 90, and 120 minutes.

[0107] 2. Perform ITT on mice: After fasting for 4 hours, subcutaneously inject insulin (0.75 U / kg body weight) and measure blood glucose at 0, 30, 60, 90 and 120 minutes.

[0108] Experiment 6: Determination of inflammatory factors and oxidative stress markers Methods: The levels of TNF-α, IL-6, MDA and SOD in serum were determined by ELISA.

[0109] step: 1. Collect mouse serum samples.

[0110] 2. Follow the instructions of the ELISA kit.

[0111] 3. Use an enzyme-labeled instrument to measure the absorbance and calculate the concentration of each indicator.

[0112] Experimental results: Table 1: In vitro antioxidant activity and α-glucosidase inhibitory activity sample DPPH clearance rate (%) ABTS clearance rate (%) α-Glucosidase inhibition rate (%) Example 1 78.5 82.3 65.7 Example 2 81.2 85.6 68.9 Example 3 83.7 87.2 71.4 Example 4 82.9 86.8 70.6 Comparative Example 1 45.3 48.1 32.5 Comparative Example 2 62.7 65.9 51.3 Comparative Example 3 58.4 61.2 47.8 Comparative Example 4 54.6 57.3 43.2 Comparative Example 5 69.8 72.5 58.1 Comparative Example 6 75.1 78.6 62.3 Table 2: Cellular glucose uptake experimental results (relative fluorescence intensity) sample 100 μg / mL 200 μg / mL 400 μg / mL Example 1 156.3 189.7 225.4 Example 2 162.8 197.5 236.9 Example 3 168.5 204.2 245.7 Example 4 165.9 201.3 241.8 Comparative Example 1 118.4 135.6 154.2 Comparative Example 2 137.2 159.8 184.5 Comparative Example 3 132.5 153.7 177.2 Comparative Example 4 128.9 148.3 170.6 Comparative Example 5 143.6 168.2 195.7 Comparative Example 6 151.7 182.4 216.9 Positive control (insulin 100 nM) 178.5 - - Table 3: Results of hypoglycemic experiment in db / db mice (week 4) sample Fasting blood glucose (mmol / L) Blood glucose 2h after meal (mmol / L) HbA1c(%) Example 1 8.2 12.5 6.4 Example 2 7.8 11.9 6.2 Example 3 7.5 11.4 6 Example 4 7.6 11.6 6.1 Comparative Example 1 12.7 18.3 8.2 Comparative Example 2 10.3 15.6 7.3 Comparative Example 3 10.8 16.2 7.5 Comparative Example 4 11.2 16.8 7.7 Comparative Example 5 9.5 14.3 6.9 Comparative Example 6 8.7 13.2 6.6 Positive control (metformin) 7.3 11.1 5.9 Model comparison 14.6 21.5 8.9 Table 4: Results of determination of inflammatory factors and oxidative stress markers sample TNF-α (pg / mL) IL-6 (pg / mL) MDA (nmol / mL) SOD (U / mL) Example 1 28.5 42.3 3.8 156.7 Example 2 26.9 39.8 3.5 162.4 Example 3 25.3 37.6 3.2 168.9 Example 4 26.1 38.5 3.3 165.7 Comparative Example 1 52.7 78.4 7.2 112.3 Comparative Example 2 41.5 61.2 5.6 132.8 Comparative Example 3 43.8 64.7 5.9 128.5 Comparative Example 4 46.2 68.3 6.3 124.1 Comparative Example 5 37.4 55.1 5 140.6 Comparative Example 6 31.8 46.9 4.2 151.3 Positive control (metformin) 24.6 36.5 3.1 171.2 Model comparison 61.3 91.7 8.5 98.6 Based on the above experimental results, the following conclusions can be drawn: 1. Synergistic antioxidant effect: Examples 1-4 showed significantly higher antioxidant activity than each single component (Comparative Examples 1 and 5). This synergistic effect may be due to the synergistic effect of polyphenols in Pinus massoniana and Pinus tabulaeformis pollen and bamboo leaf flavonoids, forming a powerful antioxidant network.

[0113] 2. Enhanced α-glucosidase inhibition: Example 3 showed the strongest α-glucosidase inhibition activity (71.4%), which was much higher than that of the single component and the non-optimized formula. This may be due to the nanoscale treatment that improves the bioavailability of the active ingredients and the synergistic effect between the multiple components.

[0114] 3. Significantly improved cellular glucose uptake capacity: Examples 1-4 performed well in promoting glucose uptake in 3T3-L1 adipocytes, even exceeding the effect of insulin at high concentrations. This result suggests that the composition of the present invention may have the effect of an insulin sensitizer.

[0115] 4. Excellent in vivo hypoglycemic effect: In the db / db mouse model, Examples 1-4 showed hypoglycemic effects comparable to metformin, especially in improving postprandial blood glucose and reducing HbA1c levels. This indicates that the composition of the present invention can not only effectively control blood glucose, but also improve long-term blood glucose management.

[0116] 5. Significant anti-inflammatory and antioxidant effects: Examples 1-4 can effectively reduce the levels of inflammatory factors (TNF-α and IL-6), while increasing the activity of the antioxidant enzyme SOD and reducing the content of lipid peroxidation product MDA. This comprehensive anti-inflammatory and antioxidant effect may help prevent and delay the occurrence of diabetic complications.

[0117] 6. Importance of raw materials from specific origins: Comparative Example 6 (using ordinary rice instead of rice from Battambang, Cambodia) also showed a certain effect, but was inferior to Examples 1 to 4 in all indicators. This verifies the key role of raw materials from specific origins in the present invention, which may be related to their unique trace element composition and active ingredient content.

[0118] 7. The key role of nanotechnology: Comparative Example 3 (non-nanoscale components) performed worse than Examples 1-4 in all tests, confirming the importance of nano-processing in improving bioavailability and enhancing drug efficacy.

[0119] 8. Necessity of optimizing the preparation process: The performance of Comparative Example 4 (non-optimized preparation process) is significantly inferior to that of Examples 1-4, indicating that the optimized preparation process in the present invention is crucial for retaining and improving the activity of each component.

[0120] In summary, the composition for treating diabetes of the present invention exhibits multiple synergistic effects and unexpected technical effects. It can not only effectively control blood sugar, but also improve insulin sensitivity, and reduce oxidative stress and inflammatory response. This multi-target, multi-mechanism mode of action provides a new idea for the comprehensive treatment of diabetes. In particular, the present invention successfully improves the bioavailability of natural active ingredients through nanotechnology and optimized preparation process, and achieves a synergistic effect that exceeds the superposition effect of a single ingredient. This innovative method may not only improve the therapeutic effect, but also reduce the dosage of the drug, thereby reducing potential side effects.

[0121] The best embodiment is embodiment 3, which performs best in all indicators. This may be because its component ratio and preparation parameters have reached the best balance, giving full play to the synergistic effect of each component.

[0122] The above description is only an embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent substitution, improvement, etc. made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.

Claims

1. A composition for treating diabetes, characterized in that: The composition comprises the following components (in parts by weight): 15-25 parts of nano-sized Masson pine pollen; 10-15 parts of nano-grade Pinus tabulaeformis pollen; 15-25 parts of bamboo leaf flavonoids-β-cyclodextrin inclusion compound; 30-40 portions of fermented rice products from Battambang, Cambodia; 10-20 parts of sea cucumber polysaccharide-polypeptide complex.

2. The composition according to claim 1, characterized in that The average particle size of the nano-scale Masson pine pollen and the nano-scale Chinese pine pollen is 50-100 nm.

3. The composition according to claim 1, characterized in that The inclusion rate of the bamboo leaf flavonoids-β-cyclodextrin inclusion compound is not less than 85%.

4. The composition according to claim 1, characterized in that The content of γ-aminobutyric acid in the rice fermentation product from the Battambang region of Cambodia is not less than 200 mg / 100g.

5. The composition according to claim 1, characterized in that The polysaccharide content in the sea cucumber polysaccharide-polypeptide complex is not less than 30%, and the polypeptide content is not less than 20%.

6. A method for preparing the composition according to any one of claims 1 to 5, characterized in that: The following steps are involved: (1) preparing nano-sized Masson pine pollen and nano-sized Pinus tabulaeformis pollen; (2) preparing bamboo leaf flavonoids-β-cyclodextrin inclusion complex; (3) Preparation of rice fermentation products from Battambang, Cambodia; (4) preparing sea cucumber polysaccharide-polypeptide complex; (5) The products obtained in steps (1) to (4) are mixed in the proportions described in claim 1.

7. The method according to claim 6, characterized in that Step (1) comprises: Firstly, the pollen of Pinus massoniana and Pinus tabulaeformis was broken by supercritical CO2 fluid technology, and the treatment conditions were pressure 20-25 MPa, temperature 40-45°C, and time 30-40 minutes. Secondly, a ZrO2 bead mill is used for nano-scale wet grinding, the grinding conditions are a rotation speed of 1500-2000 rpm, a time of 2-3 hours, and the grinding medium is deionized water; Finally, freeze drying is carried out, and the drying conditions are pre-freezing temperature of -40°C, time of 4-6 hours, main drying temperature of -20°C to 10°C, time of 24-36 hours, and post-drying temperature of 20-25°C, time of 2-4 hours.

8. The method according to claim 6, characterized in that Step (2) comprises: First, bamboo leaf powder: 70% ethanol = 1:10 (w / v), ultrasonic-assisted extraction conditions: power 500 W, frequency 40 kHz, temperature 50 ° C, time 30 minutes, filtration, and reduced pressure concentration to a paste; Secondly, the above extract was mixed with β-cyclodextrin in a molar ratio of 1:5, and stirred in purified water at 40°C for 4 hours; Then, adjust the pH to 6.8-7.2 using food-grade citric acid; Finally, spray drying is carried out, and the drying conditions are as follows: inlet temperature 150-160°C, outlet temperature 70-80°C, and atomization pressure 0.2-0.3 MPa.

9. The method according to claim 6, characterized in that Step (3) includes: First, rice from the Battambang region of Cambodia was crushed to 80-100 mesh; Secondly, rice flour and water were mixed in a ratio of 1:2 (w / v), and 1×10 lactic acid bacteria and 1×10 yeast were inoculated. 6 CFU / g; Then, ferment for 72 hours at a temperature of 30±2°C and a relative humidity of 80-85%; Again, dry at 40-45°C for 12-16 hours until the moisture content is less than 5%; Finally, the mixture was ground into 100-120 mesh using a jet mill.

10. The method according to claim 6, characterized in that Step (4) comprises: First, 0.5% food-grade papain was added to the sea cucumber and water at a ratio of 1:10 (w / v), and enzymolysis was performed at pH 6.0-6.5 and 45°C for 4 hours; Secondly, centrifuge at 5000 rpm for 15 min and collect the supernatant; Then, freeze drying was performed, and the drying conditions were pre-freezing temperature of -40°C for 4 hours, main drying temperature of -20°C to 10°C for 24 hours, and post-drying temperature of 20°C for 2 hours; Again, use a ball mill to grind to 100-120 mesh; Finally, the obtained powder was mixed with food-grade maltodextrin in a weight ratio of 1:1, stirred at 40°C for 2 hours, and then granulated using a fluidized bed granulator. The granulation conditions were: inlet temperature 50-60°C, outlet temperature 30-35°C, spray rate 8-10 mL / min, and granulation time 20-30 minutes.