Traditional Chinese medicine composition for treating diabetes and preparation method thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and relates to a traditional Chinese medicine composition for treating diabetes and a preparation method thereof. The traditional Chinese medicine composition provided by the invention comprises the following components: an aqueous extract obtained by decocting radix astragali, radix rehmanniae recen and radix puerariae according to a mass ratio of 2: 3: 1 with water and extracting; the alcohol extract is obtained by extracting coptis chinensis, folium mori and bitter gourd with ethanol according to the mass ratio of 1: 2: 1; the supercritical CO2 extract is obtained by performing supercritical CO2 extraction on salvia miltiorrhiza and polygonatum odoratum according to the mass ratio of 1: 1; the composite coating structure is formed by mixing the plant exosome and a composite additive according to the mass ratio of 5: 1. Water extraction, alcohol extraction and supercritical CO2 extraction technologies are comprehensively applied, meanwhile, a composite coating structure is introduced, the ratio of the composite coating structure to the traditional Chinese medicine extract and a preparation technology are optimized, the treatment effect of the medicine is further enhanced, and therefore an efficient and safe new scheme is provided for diabetes treatment.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and relates to a traditional Chinese medicine composition for treating diabetes and a preparation method thereof. Background Art

[0002] Diabetes is a metabolic disease characterized by chronic hyperglycemia, and its main pathological mechanisms include insulin resistance, pancreatic islet β-cell function damage, and chronic inflammatory response. Currently, the treatment with western medicines mainly relies on metformin, insulin, etc., but long-term use is prone to cause problems such as gastrointestinal reactions, hypoglycemia, and drug resistance.

[0003] Traditional Chinese medicine has shown good curative effects and low side effects in the treatment of diabetes. In the prior art, commonly used medicinal materials such as astragalus membranaceus, rehmannia glutinosa, and coptis chinensis are combined, and active ingredients are extracted through traditional processes, which have the effects of regulating blood sugar and improving insulin sensitivity. However, on the one hand, the existing traditional Chinese medicine compositions lack a targeted delivery system for pancreatic islet β-cells or inflammatory sites, and it is difficult to precisely regulate key pathological links. On the other hand, the traditional extraction process has insufficient extraction efficiency for lipophilic or thermosensitive components (such as tanshinone and flavonoids), and the drug is easily degraded in the body, resulting in limited curative effects. On the other hand, single extracts or compound prescriptions are difficult to simultaneously cover multiple mechanisms such as insulin signaling pathways, β-cell repair, and inflammation inhibition, and the treatment effects are limited.

[0004] Therefore, in the technical field of traditional Chinese medicine, it is particularly important to provide an improved traditional Chinese medicine composition for treating diabetes. Summary of the Invention

[0005] The purpose of the present invention is to provide a traditional Chinese medicine composition for treating diabetes. Through technological innovation and component optimization, the traditional Chinese medicine composition provides a new, efficient, and safe solution for the treatment of diabetes.

[0006] To achieve the above purpose, the technical solution of the present invention is as follows:

[0007] In the first aspect, the present invention provides a traditional Chinese medicine composition for treating diabetes, and the traditional Chinese medicine composition includes the following components:

[0008] An aqueous extract obtained by water decoction extraction of astragalus membranaceus, rehmannia glutinosa, and kudzu root in a mass ratio of 2:3:1;

[0009] An ethanol extract obtained by ethanol extraction of coptis chinensis, mulberry leaf, and balsam pear in a mass ratio of 1:2:1;

[0010] A supercritical CO2 extract obtained by supercritical CO2 extraction of salvia miltiorrhiza and polygonatum odoratum in a mass ratio of 1:1; and

[0011] A composite coating structure formed by mixing plant exosomes and a composite auxiliary agent in a mass ratio of 5:1.

[0012] Preferably, the plant exosomes are yam exosomes.

[0013] Preferably, the plant exosomes are not ginseng exosomes.

[0014] Preferably, the composite auxiliary agent is sodium carboxymethyl cellulose and polycaprolactone.

[0015] Preferably, the preparation method of the yam exosomes is as follows:

[0016] Cut 280 g of peeled fresh yam into pieces, add 320 mL of pre-cooled sterilized PBS (containing 0.1% β-mercaptoethanol), and intermittently crush it with a tissue crusher (30 s crushing / 20 s interval, repeat 5 times) to obtain a uniform viscous slurry. Under the condition of 4 °C, centrifuge at 4000×g for 35 min, and collect the middle layer semi-transparent colloidal liquid;

[0017] Centrifuge the middle layer semi-transparent colloidal liquid at 8000×g for 45 min (4 °C) to remove starch granules and mucilage aggregates, and take the supernatant to pass through a 0.45 μm filter membrane;

[0018] Centrifuge the filtrate at 110000×g for 70 min (4 °C). After the precipitate is resuspended with 5 mL of PBS, centrifuge at 140000×g for 80 min (4 °C). Finally, the precipitate is resuspended with 3 mL of sterile PBS and filtered and sterilized with a 0.22 μm filter to obtain yam exosomes.

[0019] Preferably, the mass ratio of sodium carboxymethyl cellulose to polycaprolactone in the composite auxiliary agent is 3:1.

[0020] Preferably, by weight, the traditional Chinese medicine composition comprises 50 - 70 parts of water extract, 20 - 30 parts of alcohol extract, 5 - 15 parts of supercritical CO2 extract, and 0.5 - 3 parts of composite coating structure.

[0021] Preferably,

[0022] The preparation method of the water extract is as follows: Mix astragalus root, rehmannia root, and kudzu root in a mass ratio of 2:3:1, soak with 10 times the amount of water for 30 min, decoct twice, each time for 1.5 h; Combine the filtrates and concentrate to a density of 1.12 - 1.18 g / cm 3 , and obtain it by spray drying;

[0023] The preparation method of the alcohol extract is as follows: Mix coptis root, mulberry leaf, and balsam pear in a mass ratio of 1:2:1, add 70% ethanol and reflux extract twice, each time for 2 h; Combine the extracts and concentrate to no alcohol smell, and obtain it by vacuum drying;

[0024] The preparation method of the supercritical CO2 extract is as follows: Salvia miltiorrhiza is dried to a water content of ≤8%, and then pulverized to 40-60 mesh; Polygonatum odoratum is processed by traditional Chinese medicine processing methods and then dried to a water content of ≤5%, and then ultrafinely pulverized to 60-80 mesh; Salvia miltiorrhiza and Polygonatum odoratum are mixed evenly according to a mass ratio of 1:1, placed in a supercritical CO2 extraction kettle for supercritical extraction, and then subjected to first-stage separation and second-stage separation. The products of the first-stage separation and the second-stage separation are combined, and the residual entrainer is removed to obtain the supercritical CO2 extract.

[0025] Preferably,

[0026] The conditions for the supercritical extraction are as follows: the pressure is 25-35 MPa, the CO2 flow rate is 25-40 kg / h, the temperature is 45-55 °C, the time is 2-4 h, and the entrainer is 5-10% anhydrous ethanol;

[0027] The conditions for the first-stage separation are as follows: the pressure is 6-8 MPa, and the temperature is 30-40 °C;

[0028] The conditions for the second-stage separation are as follows: the pressure is 3-5 MPa, and the temperature is 25-35 °C.

[0029] Preferably,

[0030] The preparation method of the composite coating structure is as follows: plant exosomes and a composite auxiliary agent are mixed according to a mass ratio of 5:1, and incubated with shaking at 37 °C for 1 h to form a composite coating structure.

[0031] It should be noted that the coating efficiency is the highest at a mass ratio of 5:1. If adjusted to other mass ratios, the plant exosomes will be exposed and easily damaged by gastric acid. Moreover, incubation with shaking at 37 °C for 1 h is to simulate the physiological temperature to promote the self-assembly of sodium carboxymethylcellulose and polycaprolactone. Too high a temperature or too long a time will cause the rupture of the plant exosome membrane, and too low a temperature or too short a time will affect the coating effect of the plant exosomes.

[0032] In a second aspect, a preparation method of the traditional Chinese medicine composition of the present invention is provided. The preparation method is as follows:

[0033] The composite coating structure and the supercritical CO2 extract are co-ground to a particle size of ≤10 μm;

[0034] They are mixed with the water extract and the alcohol extract, and 5% hydroxypropyl methylcellulose is added as a binder to granulate;

[0035] After drying, they are filled into enteric capsules.

[0036] In a third aspect, a traditional Chinese medicine preparation is provided, which is made of the traditional Chinese medicine composition of the present invention and pharmaceutically acceptable excipients.

[0037] Fourthly, there is provided an application of the traditional Chinese medicine composition described in the present invention in the preparation of a drug for reducing fasting blood glucose and / or glycated hemoglobin.

[0038] In addition, there is also provided an application of the traditional Chinese medicine composition described in the present invention in the preparation of a drug for reducing the level of inflammatory factors.

[0039] Furthermore, there is also provided an application of the traditional Chinese medicine composition described in the present invention in the preparation of a drug for improving insulin resistance and / or repairing pancreatic islet β-cell function.

[0040] Moreover, there is also provided an application of the traditional Chinese medicine composition described in the present invention in the preparation of a drug for treating diabetes.

[0041] In addition, there is also provided an application of the traditional Chinese medicine composition described in the present invention in the preparation of a drug for treating type 2 diabetes.

[0042] In the present invention, the raw materials / Chinese medicinal materials used have the following efficacy / pharmacological effects:

[0043] Astragalus membranaceus: sweet, slightly warm; attributive to the spleen and lung meridians. Tonifying qi and ascending yang, securing the exterior and stopping sweating, promoting diuresis and alleviating edema, promoting the production of body fluid and nourishing blood, astringing sores and promoting granulation.

[0044] Rehmannia glutinosa: cold in nature, sweet and bitter in taste, attributive to the heart, liver and kidney meridians, clearing heat and cooling blood, nourishing yin and promoting the production of body fluid, moistening dryness and relaxing the bowels.

[0045] Pueraria lobata: sweet, pungent, cool; attributive to the spleen, stomach and lung meridians. Relieving the muscles and expelling fever, promoting the production of body fluid and quenching thirst, ascending yang and stopping diarrhea, inducing eruption, dredging channels and collaterals.

[0046] Coptis chinensis: bitter, cold; attributive to the heart, spleen, stomach, liver, gallbladder and large intestine meridians. Clearing heat and drying dampness, purging fire and detoxifying, antibacterial and anti-inflammatory.

[0047] Mulberry leaf: sweet, bitter, cold; attributive to the lung and liver meridians. Dispelling wind and heat, clearing the liver and improving eyesight, cooling blood and stopping bleeding.

[0048] Momordica charantia: bitter, cold; attributive to the heart, spleen and lung meridians. Clearing heat and dissipating summer heat, tonifying the kidney and strengthening the spleen.

[0049] Salvia miltiorrhiza: bitter, slightly cold; attributive to the heart and liver meridians. Promoting blood circulation to remove stasis, dredging channels and relieving pain, clearing the heart and relieving vexation, cooling blood and dissipating carbuncles.

[0050] Polygonatum odoratum: sweet, slightly cold; attributive to the lung and stomach meridians. Nourishing yin and moistening dryness, promoting the production of body fluid and quenching thirst, beautifying the skin, enhancing immunity.

[0051] It should be noted that the combination of Astragalus membranaceus, Rehmannia glutinosa, and Pueraria lobata in the water extract of the present invention cannot be randomly replaced or adjusted, otherwise the reduction in fasting blood glucose will be significantly reduced. The combination of Coptis chinensis, Morus alba, and Momordica charantia in the alcohol extract of the present invention cannot be randomly replaced or adjusted, otherwise the α-glucosidase inhibition rate will be significantly decreased, and the inflammatory factor (IL-6) will rebound. Similarly, the combination of Salvia miltiorrhiza and Polygonatum odoratum in the supercritical CO2 extract of the present invention cannot be randomly replaced or adjusted, otherwise the supercritical process may not be able to effectively extract active substances or may damage the structure of active substances. The Dioscorea opposita exosomes, sodium carboxymethylcellulose, and polycaprolactone in the composite coating structure of the present invention cannot be randomly replaced or adjusted. On the one hand, because the miRNA-375 carried by Dioscorea opposita exosomes can specifically bind to the PDX-1 promoter of pancreatic islet β cells and promote insulin gene expression, replacing it with other plant exosomes (such as ginseng exosomes) will lose the targeting property. On the other hand, replacing sodium carboxymethylcellulose with other substances may not be able to stably bind to the surface charge of Dioscorea opposita exosomes, thus may lead to aggregation. On the further hand, the sustained-release skeleton formed by polycaprolactone can form "biphasic controlled release" with the hydrophilicity of sodium carboxymethylcellulose. If replaced with other substances, it may change the release kinetics (such as burst release effect).

[0052] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0053] (1) The polysaccharide and glycoside components in the water extract of the present invention act together on the insulin signaling pathway (AMPK, PI3K / Akt), improve the utilization of glucose by peripheral tissues, and protect the function of β cells. The alkaloid and polypeptide components in the alcohol extract of the present invention target key enzymes in glucose metabolism (α-glucosidase, PEPCK) and the inflammatory signaling pathway (NF-κB) to achieve dual regulation of "lowering blood sugar - anti-inflammatory". The tanshinone in the supercritical CO2 extract of the present invention and the polysaccharide of Polygonatum odoratum form a complex, which protects pancreatic islet function and delays intestinal sugar absorption through the dual effects of antioxidant and inhibiting glycosidase. The plant exosomes loaded with active ingredients in the present invention can form a stable coating structure, avoid being destroyed by gastric acid, and can be targeted and delivered to pancreatic islets and inflammatory sites. The introduced composite adjuvant can significantly enhance the overall efficacy of the traditional Chinese medicine composition, especially enhance the overall efficacy of the traditional Chinese medicine composition for patients with type 2 diabetes whose blood sugar control is still not ideal after treatment with western medicine hypoglycemic drugs or insulin. The applicant speculates that these two components may cooperate with the active ingredients in the traditional Chinese medicine composition of the present invention to jointly achieve the treatment of diabetes.

[0054] (2) Polycaprolactone (hydrophobic) and sodium carboxymethylcellulose (hydrophilic) in the present invention can form a biphasic membrane structure, reduce water penetration in a humid and hot environment, and protect Dioscorea opposita exosomes and lipophilic components (such as tanshinone) from oxidation.

[0055] (3) The supercritical process of the present invention avoids the destruction of heat-sensitive components (such as tanshinone) by high temperature, and the fat-soluble components are more likely to exist stably in the coating structure. Detailed implementation manners

[0056] The following will specifically elaborate on the present invention in combination with the detailed implementation manners and examples, and the advantages and various effects of the present invention will be presented more clearly therefrom. Those skilled in the art should understand that these detailed implementation manners and examples are used to illustrate the present invention, rather than limiting the present invention.

[0057] Below, the technical solutions of the present invention will be described in combination with examples. However, the present invention is not limited to the following examples. The experimental methods and detection methods described in each example are all conventional methods unless otherwise specified; the reagents and materials described are all commercially available unless otherwise specified.

[0058] In the embodiment of the present invention, the preparation method of the yam exosomes is as follows:

[0059] Cut 280 g of peeled fresh yam into pieces, add 320 mL of pre-cooled sterilized PBS (containing 0.1% β-mercaptoethanol), and intermittently crush it with a tissue crusher (30 s of crushing / 20 s of interval, repeat 5 times) to obtain a uniform viscous slurry. Under the condition of 4 °C, centrifuge at 4000 × g for 35 min, and collect the middle semi-transparent colloidal liquid;

[0060] Centrifuge the middle semi-transparent colloidal liquid at 8000 × g for 45 min (4 °C) to remove starch granules and mucilage aggregates, and take the supernatant to pass through a 0.45 μm filter membrane;

[0061] Centrifuge the filtrate at 110000 × g for 70 min (4 °C), resuspend the precipitate with 5 mL of PBS, then centrifuge at 140000 × g for 80 min (4 °C), finally resuspend the precipitate with 3 mL of sterile PBS, and filter and sterilize it with a 0.22 μm filter to obtain the yam exosomes.

[0062] In the embodiment of the present invention, the preparation method of the ginseng exosomes is as follows:

[0063] Take 300 g of fresh ginseng (main root part), cut it into thin slices, add 350 mL of pre-cooled PBS (containing 0.05% Triton X-100), and cycle and process it 3 times at 150 MPa with a high-pressure homogenizer to obtain a cell lysate. Under the condition of 4 °C, centrifuge at 5000 × g for 40 min, and collect the middle light yellow liquid layer;

[0064] Centrifuge the middle light yellow liquid layer at 12000 × g for 50 min (4 °C) to remove fiber fragments, and take the supernatant to pass through a 0.8 μm + 0.45 μm series filter membrane;

[0065] The filtrate was centrifuged at 130,000×g for 75 min (4 °C), the precipitate was resuspended in 4 mL of PBS, then centrifuged at 160,000×g for 85 min (4 °C), and the final precipitate was resuspended in 3 mL of PBS to obtain ginseng exosomes.

[0066] Example 1

[0067] This example provides a traditional Chinese medicine composition for treating diabetes, and the traditional Chinese medicine composition includes the following components:

[0068] An aqueous extract obtained by water decoction extraction of Astragalus membranaceus, Rehmannia glutinosa, and Pueraria lobata in a mass ratio of 2:3:1;

[0069] An ethanol extract obtained by ethanol extraction of Coptis chinensis, Morus alba leaves, and Momordica charantia in a mass ratio of 1:2:1;

[0070] A supercritical CO2 extract obtained by supercritical CO2 extraction of Salvia miltiorrhiza and Polygonatum odoratum in a mass ratio of 1:1; and

[0071] A composite coating structure formed by mixing yam exosomes and a composite auxiliary agent in a mass ratio of 5:1;

[0072] The composite auxiliary agent is sodium carboxymethylcellulose and polycaprolactone, and the mass ratio of sodium carboxymethylcellulose to polycaprolactone in the composite auxiliary agent is 3:1;

[0073] By weight, the traditional Chinese medicine composition includes 50 g of aqueous extract, 20 g of ethanol extract, 5 g of supercritical CO2 extract, and 0.5 g of composite coating structure.

[0074] Example 2

[0075] This example provides a traditional Chinese medicine composition for treating diabetes, and the traditional Chinese medicine composition includes the following components:

[0076] An aqueous extract obtained by water decoction extraction of Astragalus membranaceus, Rehmannia glutinosa, and Pueraria lobata in a mass ratio of 2:3:1;

[0077] An ethanol extract obtained by ethanol extraction of Coptis chinensis, Morus alba leaves, and Momordica charantia in a mass ratio of 1:2:1;

[0078] A supercritical CO2 extract obtained by supercritical CO2 extraction of Salvia miltiorrhiza and Polygonatum odoratum in a mass ratio of 1:1; and

[0079] A composite coating structure formed by mixing yam exosomes and a composite auxiliary agent in a mass ratio of 5:1;

[0080] The composite auxiliary agent is sodium carboxymethylcellulose and polycaprolactone, and the mass ratio of sodium carboxymethylcellulose to polycaprolactone in the composite auxiliary agent is 3:1;

[0081] By weight, the traditional Chinese medicine composition comprises 70 g of water extract, 30 g of alcohol extract, 15 g of supercritical CO2 extract and 3 g of composite coating structure.

[0082] Example 3

[0083] This example provides a traditional Chinese medicine composition for treating diabetes, and the traditional Chinese medicine composition comprises the following components:

[0084] A water extract obtained by water decocting astragalus membranaceus, rehmannia glutinosa and kudzu root in a mass ratio of 2:3:1;

[0085] An alcohol extract obtained by ethanol extracting coptis chinensis, mulberry leaf and balsam pear in a mass ratio of 1:2:1;

[0086] A supercritical CO2 extract obtained by supercritical CO2 extracting salvia miltiorrhiza and polygonatum odoratum in a mass ratio of 1:1; and

[0087] A composite coating structure formed by mixing yam exosomes and a composite auxiliary agent in a mass ratio of 5:1;

[0088] The composite auxiliary agent is sodium carboxymethylcellulose and polycaprolactone, and the mass ratio of sodium carboxymethylcellulose to polycaprolactone in the composite auxiliary agent is 3:1;

[0089] By weight, the traditional Chinese medicine composition comprises 60 g of water extract, 22 g of alcohol extract, 8 g of supercritical CO2 extract and 1.5 g of composite coating structure.

[0090] Example 4

[0091] This example provides a preparation method of the traditional Chinese medicine composition, and the preparation method is as follows:

[0092] Put the composite coating structure and the supercritical CO2 extract into a ball milling tank (zirconia balls, diameter 3 mm), with a ball-to-material ratio of 10:1, grind at 200 rpm for 30 min, and pass through a laser particle size analyzer (D90 ≤ 10 μm) to obtain a ground mixture;

[0093] Put the water extract, the alcohol extract and the ground mixture into a three-dimensional mixer (rotation speed 20 rpm, time 30 min);

[0094] Add 5% hydroxypropyl methylcellulose (dissolved in 75% ethanol) as a binder, and carry out wet granulation (sieve mesh aperture 1.0 mm, particle water content ≤ 3%);

[0095] Dry the granules in a fluidized bed at 50 °C (inlet air temperature 60 °C, outlet air temperature 40 °C), and fill them into enteric-coated capsules (each capsule contains 0.5 g ± 5%). Among them,

[0096] The preparation method of the water extract is as follows:

[0097] Weigh astragalus membranaceus, rehmannia glutinosa, and pueraria lobata according to a mass ratio of 2:3:1, remove impurities, wash, and drain the water; put the medicinal materials into a multi-functional extraction tank, add 10 times the amount (w / v) of purified water (25 - 30 °C), soak for 30 minutes, heat to boiling (100 °C), and maintain a slightly boiling state for 1.5 h, stirring once every 30 min during this period (rotation speed 50 rpm); add 8 times the amount of water (w / v) to the medicinal residues, decoct again for 1.5 h, and maintain the temperature at 98 ± 2 °C after boiling; combine the two decoction liquids, filter them roughly through a 200-mesh filter cloth, and then filter them precisely through a 0.45-μm membrane filter to remove suspended particles; transfer the filtrate to a rotary evaporator, concentrate it under the conditions of a vacuum degree of -0.08 MPa and a temperature of 80 ± 5 °C to a density of 1.12 - 1.18 g / cm3 (measured at 20 °C); spray drying [inlet air temperature 180 ± 5 °C, outlet air temperature 90 ± 3 °C, feeding speed 10 mL / min, atomization pressure 0.3 MPa, adding 2% maltodextrin (calculated based on the mass of the concentrated liquid) as a drying aid], to obtain a water extract powder with good fluidity (water content ≤ 5%, angle of repose ≤ 35°).

[0098] The preparation method of the ethanol extract is as follows:

[0099] Weigh coptis chinensis (stir-fried over a slow fire until the surface is dark yellow), mulberry leaves, and balsam pear according to a mass ratio of 1:2:1; put the medicinal materials into a reflux extraction tank, add 8 times the amount of 70% (v / v) ethanol, heat to 78 ± 2 °C, and reflux for 2 h; add 6 times the amount of 70% ethanol to the medicinal residues, repeat refluxing for 2 h, and combine the two extraction liquids; vacuum concentrate the extraction liquid (50 °C, -0.06 MPa) until there is no alcohol smell (GC detects that the ethanol residue ≤ 0.1%); dry the concentrated liquid in a vacuum drying oven at 60 °C (vacuum degree -0.1 MPa, time 48 h), pulverize it through an 80-mesh sieve to obtain an ethanol extract powder (water content ≤ 5%).

[0100] The preparation method of the supercritical CO2 extract is as follows:

[0101] Dry salvia miltiorrhiza until the water content ≤ 8%, and pulverize it to 40 - 60 meshes; dry polygonatum odoratum after processing until the water content ≤ 5%, and superfine pulverize it to 60 - 80 meshes; mix salvia miltiorrhiza and polygonatum odoratum evenly according to a mass ratio of 1:1, put them into a supercritical CO2 extraction kettle for supercritical extraction, then perform primary separation and secondary separation, combine the products of primary separation and secondary separation, and remove the residual entrainer to obtain the supercritical CO2 extract.

[0102] The conditions for supercritical extraction are: the pressure is 25 - 35 MPa, the CO2 flow rate is 25 - 40 kg / h, the temperature is 45 - 55 °C, the time is 2 - 4 h, and the entrainer is 5 - 10% absolute ethanol;

[0103] The conditions for primary separation are: the pressure is 6 - 8 MPa, and the temperature is 30 - 40 °C;

[0104] The conditions for secondary separation are: pressure of 3 - 5 MPa and temperature of 25 - 35 °C.

[0105] The preparation method of the composite coating structure is as follows:

[0106] Mix sodium carboxymethylcellulose and polycaprolactone in a mass ratio of 3:1, dissolve them in 0.1 M acetate buffer (pH 5.5), and ultrasonically disperse (40 kHz, 30 min) to obtain a homogeneous colloid (i.e., the composite auxiliary agent); mix yam exosomes and the composite auxiliary agent in a mass ratio of 5:1 (10 mg of yam exosomes and 2 mg of the composite auxiliary agent), incubate in a shaker at 37 °C (150 rpm) for 1 h; remove the free auxiliary agent by dialysis (MWCO 100 kDa), and freeze-dry to obtain the composite coating structure powder (coating efficiency ≥ 90%, determined by dynamic light scattering method).

[0107] Comparative Example 1

[0108] This comparative example is the same as Example 3, except that yam exosomes are replaced with ginseng exosomes.

[0109] Comparative Example 2

[0110] This comparative example is the same as Example 3, except that danshen and polygonatum odoratum are mixed in a mass ratio of 1:1, soaked in 10 times the amount of water (w / v) for 30 min, heated to boiling, maintained for 1.5 h, and filtered to obtain the first extraction solution; the medicinal residues are added with 8 times the amount of water (w / v), decocted again for 1.5 h, and the two filtrates are filtered and combined; the combined filtrate is concentrated at 80 °C to a density of 1.10 - 1.15 g / cm3 to obtain a concentrated solution; the concentrated solution is spray-dried (inlet air temperature is 180 °C, outlet air temperature is 90 °C, and 2% maltodextrin is added as a drying aid); after drying, the dried product is collected to obtain the water extract of danshen and polygonatum odoratum.

[0111] Comparative Example 3

[0112] This comparative example is the same as Example 3, except that polycaprolactone is not added.

[0113] Effect verification

[0114] I. Experimental animals and modeling methods

[0115] 1. Experimental animals

[0116] Healthy male SD rats without specific pathogens (body weight 180 - 220 g, SPF level) are selected and raised in individually ventilated cages at a temperature of 22 ± 1 °C, humidity of 50 ± 5%, and a 12 h / 12 h light-dark cycle. There are a total of 90 rats, which are randomly divided into the following 9 groups, with 10 rats in each group.

[0117] Blank control group: Fed with normal feed, not modeled, and given normal saline.

[0118] Model group: fed with high-fat and high-sugar diet + chemically induced diabetes, given normal saline.

[0119] Positive control group: metformin group (150 mg / kg / d, dissolved in 0.5% sodium carboxymethylcellulose solution).

[0120] Experimental group (Examples 1-3): diabetic model rats, given the traditional Chinese medicine compositions of Examples 1, 2, and 3 respectively (dose: 1.5 g of crude drug / kg / d).

[0121] Comparison group (Comparative Examples 1-3): diabetic model rats, given the traditional Chinese medicine compositions of Comparative Examples 1, 2, and 3 respectively (dose: 1.5 g of crude drug / kg / d).

[0122] 2. Establishment of diabetic model

[0123] High-fat and high-sugar diet: 52% ordinary feed, 12% coconut oil, 20% fructose, 2.2% cholesterol, 0.8% sodium cholate, 10% casein, 2% vitamin-mineral premix, 1% cellulose; after feeding for 3 weeks, combined with chemical induction.

[0124] Chemical induction: after fasting for 16 h, intraperitoneally inject freshly prepared STZ (Sigma, product number S0130) solution (35 mg / kg, dissolved in 0.1 M citric acid-sodium citrate buffer, pH 4.6, operate under ice bath and avoid light).

[0125] Blood glucose verification: collect blood from the tail vein twice at 48 h and 72 h after injection, rats with blood glucose ≥ 16.7 mmol / L and lasting for 3 days are included in the experiment, and self-healing individuals (blood glucose fluctuation < 15%) are excluded.

[0126] 3. Adaptive swallowing treatment

[0127] Ten days before modeling, custom-made silicone simulation capsules (size: length 10 mm × diameter 4 mm, containing 0.1% beef extract to induce licking) are used regularly every day, combined with operant conditioning training (give 0.5% sucrose water reward after correct swallowing);

[0128] Before the formal experiment, randomly select 5 rats to orally take enteric-coated capsules containing 1% sodium fluorescein, anesthetize and dissect them 2 h later, and observe the intestinal fluorescence distribution under ultraviolet light to confirm the capsule disintegration site (below the duodenum).

[0129] 4. Administration scheme

[0130] After successful model establishment, each group was continuously administered drugs for 6 weeks, and gavage was performed at a fixed time every day (for example, from 9:00 to 10:00). Calculated by the crude drug amount, it was converted into the capsule loading amount according to 1.5 g / kg (the loading amount per capsule is 0.5 g ± 5%), and the dosage was adjusted according to the real-time body weight.

[0131] 5. Quality control

[0132] The dissolution test was performed on each batch of capsules (enteric-coated capsule standard in the 2020 edition of the Chinese Pharmacopoeia) to ensure that the release rate of the active ingredient in simulated intestinal fluid was ≥85% within 2 h.

[0133] II. Experimental detection indicators

[0134] 1. Blood glucose level

[0135] Fasting blood glucose (FBG): Before drug administration and 4 weeks after drug administration, the tail vein blood glucose was measured after 4 h of fasting (blood glucose meter).

[0136] Glycated hemoglobin (HbA1c): 4 weeks after drug administration, blood was collected from the abdominal aorta and detected by high performance liquid chromatography.

[0137] 2. Insulin sensitivity and β-cell function

[0138] Fasting insulin (FINS): Detected by ELISA method.

[0139] HOMA-IR index: Calculation formula: HOMA-IR = (FBG × FINS) / 22.5.

[0140] Islet β-cell area: Pancreatic tissue sections (HE staining), and the proportion of β-cells was calculated under the microscope.

[0141] 3. Inflammatory factor levels

[0142] IL-6, TNF-α: The content of inflammatory factors in serum was detected by ELISA method.

[0143] III. Experimental results

[0144] The blood glucose and HbA1c levels of rats in each group ( n = 10) are shown in Table 1; the results of insulin sensitivity and β-cell function ( n = 10) are shown in Table 2; the results of inflammatory factors of rats in each group ( n = 10) are shown in Table 3.

[0145] Table 1 Blood glucose and HbA1c levels of rats in each group

[0146]

[0147]

[0148] As can be seen from Table 1, both FBG and HbA1c in the experimental group (Examples 1-3) were significantly lower than those in the model group and the positive control group, and Example 3 had the best effect, indicating that the optimized ratio of traditional Chinese medicine composition could more effectively control blood glucose. The effect of Comparative Example 1 was weaker than that of the experimental group, suggesting that the targeted delivery of yam exosomes to active ingredients was specific. The effects of Comparative Example 2 and Comparative Example 3 were the worst, indicating that supercritical CO2 extraction and composite coating structure were crucial for maintaining the drug effect.

[0149] Table 2 Results of insulin sensitivity and β-cell function

[0150] Group HOMA-IR index β-cell area (%) Blank control group 1.2±0.1 3.8±0.2 Model group 6.8±0.5 0.9±0.1 Positive control group 3.5±0.3 2.0±0.2 Example 1 2.8±0.2 2.5±0.3 Example 2 2.4±0.2 2.8±0.3 Example 3 1.9±0.1 3.2±0.2 Comparative example 1 4.0±0.3 1.8±0.2 Comparative example 2 5.5±0.4 1.2±0.1 Comparative example 3 4.8±0.3 1.5±0.1

[0151] Table 3 Results of inflammatory factors in rats of each group

[0152]

[0153]

[0154] As can be seen from Table 2 and Table 3, the experimental group (Examples 1-3) could reduce inflammatory factors (IL-6, TNF-α) and HOMA-IR, and restore the β-cell area. Among them, Example 3 could significantly reduce inflammatory factors (IL-6, TNF-α) and HOMA-IR, and restore the β-cell area to near the normal level, indicating that it could play a good therapeutic role by anti-inflammation and improving insulin sensitivity. In Comparative Example 1, the targeting decreased due to the replacement of exosomes. In Comparative Example 2, the effective components were lost due to the change of extraction method. In Comparative Example 3, the stability of the coating structure decreased due to the lack of polycaprolactone (the mechanical properties of sodium carboxymethylcellulose alone forming a film were poor), and the active components were prematurely released and destroyed by gastric acid. The effects of the three in the control group were significantly inferior to those of the experimental group.

[0155] In addition, the examples of the present invention also verified the stability [enteric-coated capsule products of Examples 1, 2, 3, Comparative Example 1, Comparative Example 2, and Comparative Example 3 (3 parallel samples for each batch)], and the accelerated conditions were set: 40°C ± 2°C / 75% RH ± 5%, lasting for 6 months. It was confirmed by the accelerated test data that the composite coating structure and process of the experimental group (Examples 1-3) could significantly improve the stability (content change rate < 5%). In the control group (Comparative Example 1-Comparative Example 3) (change rate > 5%, even > 15%), the stability was significantly inferior to that of Example 3 in the experimental group due to technical defects. Specifically, in Comparative Example 1, it might be because of the poor compatibility between ginseng exosomes and composite auxiliaries, and the coating structure was prone to collapse under humid and hot conditions, resulting in the leakage of active ingredients. In Comparative Example 2, it might be because the extraction rate of tanshinone was low due to the water extraction method, and the uncoated lipophilic components were more prone to oxidation and degradation. In Comparative Example 3, it might be because of the lack of hydrophobic support of polycaprolactone, and the sodium carboxymethylcellulose membrane swelled and ruptured under high temperature and high humidity, accelerating the degradation of components.

[0156] It should be understood that the present invention disclosed is not limited to the specific methods, schemes and substances described, as these can vary. It should also be understood that the terms used herein are for the purpose of describing specific embodiments only and are not intended to limit the scope of the present invention, which is limited only by the appended claims.

Claims

1. A traditional Chinese medicine composition for treating diabetes, characterized in that, The traditional Chinese medicine composition comprises the following components: An aqueous extract obtained by water decocting astragalus membranaceus, rehmannia glutinosa, and kudzu root in a mass ratio of 2:3:1; An ethanol extract obtained by ethanol extracting coptis chinensis, mulberry leaves, and balsam pear in a mass ratio of 1:2:1; A supercritical CO2 extract obtained by supercritical CO2 extracting salvia miltiorrhiza and polygonatum odoratum in a mass ratio of 1:1; and A composite coating structure formed by mixing plant exosomes and a composite auxiliary agent in a mass ratio of 5:

1.

2. The traditional Chinese medicine composition according to claim 1, wherein The plant exosomes are yam exosomes.

3. The traditional Chinese medicine composition according to claim 1, wherein The composite auxiliary agent is sodium carboxymethyl cellulose and polycaprolactone.

4. The traditional Chinese medicine composition according to claim 1, wherein By weight, the traditional Chinese medicine composition comprises 50 - 70 parts of the aqueous extract, 20 - 30 parts of the ethanol extract, 5 - 15 parts of the supercritical CO2 extract, and 0.5 - 3 parts of the composite coating structure.

5. The traditional Chinese medicine composition according to claim 1, wherein The preparation method of the water extract is as follows: Astragalus membranaceus, Rehmannia glutinosa and Pueraria lobata are mixed in a mass ratio of 2:3:1, soaked in 10 times the amount of water for 30 min, decocted twice, each time for 1.5 h; the filtrates are combined and concentrated to a density of 1.12 - 1.18 g / cm 3 , and obtained by spray drying; The preparation method of the ethanol extract is as follows: Mix coptis chinensis, mulberry leaves, and balsam pear in a mass ratio of 1:2:1, add 70% ethanol and reflux extract twice, each time for 2 hours; Combine the extraction solutions and concentrate to a non-alcoholic taste, and obtain by vacuum drying; The preparation method of the supercritical CO2 extract is as follows: Dry salvia miltiorrhiza to a water content ≤ 8%, and crush it to 40 - 60 meshes; After processing polygonatum odoratum, dry it to a water content ≤ 5%, and ultramicro crush it to 60 - 80 meshes; Mix salvia miltiorrhiza and polygonatum odoratum evenly in a mass ratio of 1:1, place them in a supercritical CO2 extraction kettle for supercritical extraction, then perform primary separation and secondary separation, combine the primary separation and secondary separation products, and remove the residual entrainer to obtain the supercritical CO2 extract.

6. The traditional Chinese medicine composition according to claim 5, wherein The conditions for the supercritical extraction are: the pressure is 25 - 35 MPa, the CO2 flow rate is 25 - 40 kg / h, the temperature is 45 - 55 °C, the time is 2 - 4 hours, and the entrainer is 5 - 10% anhydrous ethanol; The conditions for the primary separation are: the pressure is 6 - 8 MPa, and the temperature is 30 - 40 °C; The conditions for the secondary separation are: the pressure is 3 - 5 MPa, and the temperature is 25 - 35 °C.

7. The traditional Chinese medicine composition according to claim 1, wherein The preparation method of the composite coating structure is as follows: Mix plant exosomes and a composite auxiliary agent in a mass ratio of 5:1, and incubate with shaking at 37 °C for 1 hour to form a composite coating structure.

8. The preparation method of the traditional Chinese medicine composition according to any one of claims 1-7, characterized in that, The preparation method is as follows: Co-grind the composite coating structure and the supercritical CO2 extract until the particle size ≤ 10 μm; Mix with the aqueous extract and the ethanol extract, and add 5% hydroxypropyl methylcellulose as a binder to granulate; After drying, fill it into enteric capsules.

9. Use of the traditional Chinese medicine composition according to any one of claims 1 - 7 in the preparation of a medicament for treating diabetes.

10. Use of the traditional Chinese medicine composition according to any one of claims 1 - 7 in the preparation of a medicament for reducing fasting blood glucose and / or glycated hemoglobin.

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