A composition, preparation and preparation method for regulating blood lipid levels

Abstract

The invention relates to the field of medical technology, and in particular to a composition, a preparation and a preparation method for regulating blood lipid levels. The invention selects bamboo leaf flavonoids, sugarcane polyphenols, phytosterols, natto powder, water-insoluble dietary fiber and kudzu root powder, which are both medicinal and edible materials with small side effects, as the blood lipid regulating composition. The bamboo leaf flavonoids, sugarcane polyphenols and phytosterols are firstly prepared into drug-loaded nanoparticles with good stability, high drug loading amount and temperature-sensitive controlled release drug delivery; and then the nanoparticles are scientifically matched with natto powder, water-insoluble dietary fiber and kudzu root powder to inhibit cholesterol synthesis, increase HDL-C content and reduce LDL-CTC and TG content, thereby playing the role of inhibiting cholesterol synthesis and regulating blood lipids.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technologies, and particularly relates to a composition, a preparation and a preparation method for regulating blood lipid levels. Background Art

[0002] With the improvement of people's living standards, the prevalence of metabolic syndrome caused by abnormal blood lipids, blood glucose, etc. has increased. A large number of studies have shown that hyperlipidemia is an important factor in many diseases such as obesity, hypertension, coronary heart disease or diabetes. Therefore, people have been actively exploring drugs for treating hyperlipidemia for many years.

[0003] There are many drugs for lowering blood lipids on the market now, such as statins, niacin, fibrates, etc. However, these drugs have a high probability of bringing some side effects to patients, including rhabdomyolysis, liver dysfunction, thrombocytopenia, etc. Traditional Chinese medicine compound prescriptions have become an important means for preventing blood lipid increase and assisting in lowering blood lipids today due to their advantages of people-oriented, comprehensive conditioning, and consolidating the foundation and cultivating the vitality. Developing traditional Chinese medicine compound prescriptions into health foods for regulating blood lipids and formulating them into dosage forms acceptable to the public to make up for the disadvantages of large side effects of western medicines in lowering blood lipids will help improve the blood lipid status of the people in our country. At present, most traditional Chinese medicine compound drugs are developed for hyperlipidemia, aiming to quickly lower blood lipids, and cannot eliminate the internal causes inducing hyperlipidemia. When the drugs are taken in excess, symptoms of hypolipidemia will be induced, and when the blood lipids are too low, symptoms such as reduced liver function and endocrine disorders will also occur. And limited by the physicochemical properties, traditional Chinese medicine oral preparations will degrade in the acidic environment of the stomach, resulting in the loss of effective active ingredients and being unable to enter the intestine for effective absorption and utilization by the human body, with poor bioavailability and thus poor oral treatment effects. Therefore, under the guidance of traditional Chinese medicine theory, selecting edible and medicinal materials to effectively control blood lipid abnormalities and combining with pharmaceutical technology to improve the blood drug concentration and blood exposure of oral administration have important clinical significance. Summary of the Invention

[0004] In order to overcome the deficiencies of the above technical defects, the present invention gives full play to the advantages of traditional Chinese medicine health preservation, rationally combines edible and medicinal materials with small side effects, and provides a composition, a preparation and a preparation method for regulating blood lipid levels, which have the effects of inhibiting cholesterol synthesis, increasing content, and reducing , , content, so as to play a role in inhibiting cholesterol synthesis and regulating blood lipids.

[0005] One of the objectives of the present invention is to provide a composition for regulating blood lipid levels. The key lies in: including the following raw materials in parts by mass: 280 - 310 parts of drug-loaded nanoparticles, 4 - 5 parts of natto powder, 150 - 200 parts of insoluble dietary fiber, and 225 - 300 parts of kudzu root powder; among them, the drug-loaded nanoparticles are formed by loading drugs onto pullulan-grafted poly-(lactic acid-glycolic acid) to form drug-loaded nanoparticles with a particle size of 90 - 100 nm, and the drug loading of the drug-loaded nanoparticles is 57.8 ± 0.5%; the drug is 53 - 65%wt of bamboo leaf flavonoids, 23 - 42%wt of sugarcane polyphenols, and 7 - 11% of phytosterols.

[0006] Medicinal and edible traditional Chinese medicines refer to animals, plants, minerals and their processed products that can be used both as food and as medicine, and have the characteristics of a wide dosage range and low toxicity and side effects.

[0007] Bamboo leaf flavonoids: inhibit fatty acid synthase to reduce the generation of endogenous fatty acids and promote the β-oxidation of fatty acids;

[0008] Sugarcane polyphenols: can simultaneously inhibit α-amylase and α-glucosidase, perfectly inhibit the decomposition and absorption of starch, polysaccharides, disaccharides, fructose, and glucose, and can intercept blood glucose changes caused by various sugars;

[0009] Phytosterols: reduce cholesterol absorption by reducing the expression of NPC1L61 in the small intestine;

[0010] Natto powder: significantly reduce and increase the ratio. Soybean fiber can directly interfere with the absorption of cholesterol in the intestine or the reabsorption of bile in the intestine, significantly increasing the excretion of bile acids in the human body, thus playing an important role in reducing blood lipids; at the same time, Bacillus natto is not affected by the strong acid of gastric juice and can quickly colonize in the intestine; the pyridine dicarboxylic acid produced by Bacillus natto effectively kills and inhibits harmful bacteria and viruses in the intestine, making the intestinal flora reach a dynamic balance beneficial to health, thereby regulating blood lipid balance by regulating intestinal microorganisms. At the same time, nattokinase can also dissolve thrombus, fat, etc. in capillaries, and can also achieve a good effect of reducing blood lipids;

[0011] Insoluble dietary fiber: highly adsorb nitrite and cholesterol, reducing the human body's absorption of cholesterol and other substances;

[0012] Puerarin in kudzu root powder can promote the conversion of cholesterol into bile acids and excrete them out of the body, and puerarin flavone can inhibit fatty acid synthase to reduce the generation of endogenous fatty acids and promote the β-oxidation of fatty acids;

[0013] Flavonoids, polyphenols and sterol compounds can only be effectively absorbed and utilized by the human body after entering the intestine. Increasing the hydrophilicity of flavonoid compounds can improve the bioavailability of absorption in the intestine. The inventor selected poly-(lactic acid-glycolic acid)-modified pullulan as a carrier, and self-assembled with flavonoids, polyphenols and sterol compounds to form drug-loaded nanoparticles with a particle size of 90-100 nm. The drug loading of the drug-loaded nanoparticles was 57.8±0.5%, and the encapsulation efficiency was 81.6±2.1%. It had a high encapsulation efficiency and drug loading, prolonged the circulation time of the drug in the blood, and improved the bioavailability of the drug;

[0014] Further, the drug-loaded nanoparticles are prepared by the following method: Dissolve the drug and pullulan grafted with poly-(lactic acid-glycolic acid) with a mass ratio of 1:(9-12) in methanol, and heat under reduced pressure at 37°C to obtain a composite film; Mix the composite film and the hydration solution, stir and hydrate in an ice-water bath for 10-20 min to obtain a suspension; Centrifuge and separate the suspension, take the supernatant and filter it, and filter through a microporous filter membrane to remove the unencapsulated raw materials to prepare a nanoparticle solution, and lyophilize the obtained nanoparticle solution to obtain the product.

[0015] During the process of the inventor preparing the drug-loaded nanoparticles by the thin film hydration method, as methanol is removed, the interaction between the hydrophobic ends of the drug and poly-(lactic acid-glycolic acid)-modified pullulan gradually strengthens, and finally a composite film of the two is formed. When the aqueous phase is added, due to the strong interaction between the two that has been formed, the two together serve as the hydrophobic segment and are wrapped into the core of the nanoparticle, and the hydrophilic outer shell avoids the contact between the drug and the aqueous environment and reduces the precipitation of the drug. The lower critical solution temperature of poly-(lactic acid-glycolic acid)-modified pullulan in the aqueous phase is about 40°C. As the temperature rises, secondary aggregation will occur to form new aggregates with a larger volume, and when the temperature drops again, the new aggregates will disassemble and return to the original small aggregates;

[0016] Since the thin film hydration method is not affected by the external environment, it is most conducive to the formation of a strong interaction between the drug and poly-(lactic acid-glycolic acid)-modified pullulan, and the formation of a strong interaction between the two is more conducive to the compression of the hydrophobic core of the nanoparticles, so that the prepared nanoparticles have a smaller particle size and a higher encapsulation efficiency.

[0017] Further, the hydration solution is a phosphate buffer solution with a mass fraction of 3-10%; preferably a phosphate buffer solution with a mass fraction of 5-8%; more preferably a phosphate buffer solution with a mass fraction of 5.5%.

[0018] The inventor uses phosphate buffer solution as the hydration solution, which can increase the viscosity of the system, and can interact with the hydrophilic end of poly-(lactic acid-glycolic acid)-modified pullulan and can change the charge property on the surface of the nanoparticles. Therefore, the prepared drug-loaded nanoparticles have small particle size and high encapsulation rate, which are significantly superior to other conventional stabilizers such as TPGS and PIuronic.

[0019] Furthermore, the pullulan grafted with poly-(lactic acid-glycolic acid) is prepared by the following method: DMSO is added to pullulan polysaccharide, and after heating and stirring to dissolve, lactide and glycolide are added. Under nitrogen protection, after mixing evenly, stannous octoate is added, and microwave reaction is carried out at 80 °C for 3-8 min. Among them, the molar ratio of stannous octoate to lactide is (0.06-0.24):100, the molar ratio of lactide to the hydroxyl group of pullulan polysaccharide is (60-85):100, and the molar ratio of glycolide to lactide is (5-10):100.

[0020] The inventor synthesizes pullulan grafted with poly-(lactic acid-glycolic acid) with the required lower critical solution temperature by adjusting the ratio of lactide and glycolide and the ratio of ester monomer and pullulan polysaccharide, so as to have the characteristics of stimulus-responsive controlled release, in order to achieve the application purpose of sustained-release drug delivery.

[0021] Furthermore, the water-insoluble dietary fiber is selected from one or more of oat fiber, bamboo shoot fiber, wheat fiber,

[0022] corn fiber, soybean fiber, and pea fiber.

[0023] The second object of the present invention is to provide an enteric-coated preparation, the key lies in: including the above-mentioned composition and enteric coating; the release amount of the enteric-coated preparation in artificial gastric juice within 4 hours < 10%, and the release amount in artificial intestinal juice within 4 hours > 90%.

[0024] The enteric-coated preparation of the present invention has the advantages of almost no release in the stomach, stable release in the intestinal environment, high bioavailability, etc., greatly increasing the compliance of patients taking the medicine, and can meet the existing clinical medication needs.

[0025] Preferably, the release amount of the enteric-coated preparation in artificial gastric juice within 4 hours does not exceed 5%, and the release amount in artificial intestinal juice within 4 hours > 90%.

[0026] Among them, the release amount is reflected as dissolution data, and the dissolution determination method (basket method) specified in the 2015 edition of the Pharmacopoeia is used in the present invention.

[0027] Further, based on 100 parts by mass, the enteric coating comprises 60 - 75 parts of film-forming material, 8 - 12 parts of plasticizer, and the balance is solvent; preferably, the film-forming material is 65 - 70 parts, and the plasticizer is 10 - 12 parts; more preferably, the film-forming material is 68 parts, and the plasticizer is 11 parts.

[0028] Further, the film-forming material is selected from one or more of polyacrylic resin, hydroxypropyl methylcellulose acetate succinate, cellulose acetate phthalate, ethyl cellulose, polyvinyl alcohol phthalate, enteric Opadry, carboxymethyl ethyl cellulose, polyvinyl acetate phthalate, and diketopiperazine polymer;

[0029] The plasticizer is preferably one or more of polyethylene glycol, propylene glycol, triethyl citrate, diethyl phthalate, tributyl citrate, and glycerol monostearate and distearate;

[0030] The third object of the present invention is a preparation method of an enteric preparation, which is characterized by comprising the following steps: fully stirring and mixing the drug-loaded nanoparticles, natto powder, water-insoluble dietary fiber, kudzu root powder with an adsorbent to form an inclusion powder with a particle size of 0.6 - 1.5 mm; performing enteric coating on the inclusion powder in a fluidized bed; preferably, the parameters of the fluidized bed are: controlling the inlet air volume to be 270 - 330 m 3 / h, the drug loading rate is 6 g - 15 g / min, the inlet air temperature is 50 - 60 °C, the material temperature is 32 - 35 °C, and the atomization pressure is 0.1 MPa - 0.3 MPa; the suspension stirring speed is 500 - 750 rpm, and the stirring time is 25 - 35 min.

[0031] Further, the mass ratio of the enteric coating to the inclusion powder is (0.15 - 0.25):1; preferably (0.18 - 0.22):1; more preferably 0.2:1.

[0032] In a specific embodiment, the adsorbent is one or more of silica, cyclodextrin, lactose, mannitol, calcium hydrogen phosphate, calcium sulfate, microcrystalline cellulose, and acrylic resin.

[0033] The present invention adopts a preparation design of multi-drug release units with pellets coated with an adsorbent, avoiding the risk of sudden release or delayed release of a single drug release unit.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] (1) The composition for regulating blood lipid levels provided by the present invention, based on the theory of traditional Chinese medicine with homology of medicine and food, first prepares drug-loaded nanoparticles with good stability, high drug loading capacity, and temperature-sensitive controlled release for drug delivery from bamboo leaf flavonoids, sugarcane polyphenols, and phytosterols; then scientifically formulates them with natto powder, water-insoluble dietary fiber, and kudzu root powder, which can inhibit cholesterol synthesis, increase content, and reduce , content, thereby playing a role in inhibiting cholesterol synthesis and regulating blood lipid levels;

[0036] (2) The enteric-coated preparation provided by the present invention adopts the preparation design of multi-drug release units with pellets coated, avoiding the risks of sudden release or delayed release of single drug release units, having the advantages of almost no release in the stomach, stable release in the intestinal environment, high bioavailability, etc., which can meet the existing clinical medication needs, and the preparation process is simple.

[0037] (3) The prescription and process parameter range of the present invention are relatively large, which is easy to verify the production process and meets the actual production requirements. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 is a figure of the drug-loaded nanoparticles prepared by the present invention;

[0039] Figure 2 is a figure of the pullulan-grafted poly-(lactic acid-glycolic acid) prepared by the present invention;

[0040] Figure 3 is a figure of the pullulan-grafted poly-(lactic acid-glycolic acid) prepared by the present invention;

[0041] Figure 4 is the particle size distribution diagram of the pullulan-grafted poly-(lactic acid-glycolic acid) prepared by the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0042] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be described in detail below in conjunction with specific embodiments. The experimental methods without specific conditions noted in the following examples are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight. Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to the described content can be applied to the present invention. The preferred methods and materials described herein are for illustrative purposes only.

[0043] The endpoints and any values within the ranges disclosed herein are not limited to the exact ranges or values. These ranges or values should be understood to include values close to these ranges or values. For numerical ranges, the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be considered to be specifically disclosed herein.

[0044] According to the present invention, an enteric-coated preparation with a sustained-release effect adopts a preparation design of multi-drug release units with pellets coated. The drug-loaded nanoparticles, natto powder, water-insoluble dietary fiber, kudzu powder and adsorbent are fully stirred and mixed to prepare an inclusion powder with a particle size of 0.6 - 1.5 mm; the inclusion powder is prepared by coating an enteric coating layer in a fluidized bed, avoiding the risk of sudden release or delayed release of single drug release units. The release amount of the enteric-coated preparation in artificial gastric juice within 4 hours is < 10%, and the release amount in artificial intestinal juice within 4 hours is > 90%;

[0045] According to the present invention, the parameters of the fluidized bed are: the incoming air volume is controlled to be 270 - 330 m 3 / h, the feeding rate is 6 g - 15 g / min, the incoming air temperature is 50 - 60 °C, the material temperature is 32 - 35 °C, and the atomization pressure is 0.1 MPa - 0.3 MPa; the suspension stirring speed is 500 - 750 rpm, and the stirring time is 25 - 35 min;

[0046] According to the present invention, the mass ratio of the enteric coating to the inclusion powder is (0.15 - 0.25):1; in some specific embodiments, the mass ratio of the enteric coating to the inclusion powder is (0.18 - 0.22):1; in some other specific embodiments, the mass ratio of the enteric coating to the inclusion powder is 0.2:1;

[0047] According to the present invention, the water-insoluble dietary fiber is selected from one or more of oat fiber, bamboo shoot fiber, wheat fiber, corn fiber, soybean fiber, and pea fiber;

[0048] According to the present invention, based on 100 parts by mass, the enteric coating includes 60 - 75 parts of film-forming material, 8 - 12 parts of plasticizer, and the balance is solvent;

[0049] According to the present invention, the adsorbent is one or more of silica, cyclodextrin, lactose, mannitol, calcium hydrogen phosphate, calcium sulfate, microcrystalline cellulose, and acrylic resin. In actual operation, it can be selected according to the situation, and will not be elaborated one by one here.

[0050] According to the present invention, the film-forming material is selected from one or more of polyacrylic resin, hydroxypropylmethylcellulose acetate succinate, cellulose acetate phthalate, ethyl cellulose, polyvinyl alcohol phthalate, enteric Opadry, carboxymethyl ethyl cellulose, polyvinyl acetate phthalate, and diketopiperazine polymer; in actual operation, selection can be made according to circumstances, and details will not be elaborated herein one by one.

[0051] According to the present invention, the solvent is a conventional substance in the art, preferably an aqueous ethanol solution with a volume concentration of 95%.

[0052] According to the present invention, the plasticizer is preferably one or more of polyethylene glycol, propylene glycol, triethyl citrate, diethyl phthalate, tributyl citrate, and glycerol monostearate and glycerol distearate; in actual operation, selection can be made according to circumstances, and details will not be elaborated herein one by one.

[0053] According to the present invention, the drug-loaded nanoparticles are prepared by the following method: Dissolve the drug and pullulan-grafted poly-(lactic acid-glycolic acid) with a mass ratio of 1:(9 - 12) in methanol, and heat under reduced pressure at 37 °C to obtain a composite film; Mix the composite film and a phosphate buffer solution with a mass fraction of 3 - 10%, and stir and hydrate in an ice-water bath for 10 - 20 min to obtain a suspension; Centrifuge and separate the suspension, take the supernatant and filter it, and filter through a microporous membrane to remove the unloaded raw materials to prepare a nanoparticle solution, and lyophilize the obtained nanoparticle solution to obtain the product.

[0054] According to the present invention, the drug loading of the drug-loaded nanoparticles is 57.8 ± 0.5%; the drug includes 53 - 65%wt of bamboo leaf flavonoids, 23 - 42%wt of sugarcane polyphenols, and 7 - 11% of phytosterols;

[0055] According to the present invention, the pullulan-grafted poly-(lactic acid-glycolic acid) is prepared by the following method: Add , heat and stir to dissolve, then add lactide and glycolide, under nitrogen protection, mix evenly and then add stannous octoate, and carry out microwave reaction at 80 °C for 3 - 8 min, wherein the molar ratio of stannous octoate to lactide is (0.06 - 0.24):100, the molar ratio of lactide to the hydroxyl group of pullulan polysaccharide is (60 - 85):100, and the molar ratio of glycolide to lactide = (5 - 10):100.

[0056] The technical solutions of the present invention will be further described in detail below with specific examples and accompanying drawings. It should be understood that the following examples are only used to explain the present invention and are not used to limit the present invention.

[0057] Example 1 Preparation of Pullulan-Grafted Poly-(Lactic Acid-Glycolic Acid)

[0058] Add After heating, stirring and dissolving, lactide and glycolide are added. Under nitrogen protection, after mixing evenly, stannous octoate is added. At 80 °C, microwave reaction is carried out for 3 - 8 min. Among them, the molar ratio of stannous octoate to lactide is 0.2:100, the molar ratio of lactide to the hydroxyl groups of pullulan polysaccharide is 80:100, and the molar ratio of glycolide to lactide is 8:100. The results measured by dynamic light scattering show that the lowest lower critical solution temperature is 40 °C.

[0059] Figure 2-3 For pullulan-grafted poly-(lactic acid-glycolic acid) and Figure, Figure 2 As can be seen, taking pullulan polysaccharide as a control, the characteristic spectral bands of pullulan polysaccharide (3400 - 3200 cm -1 ) are the stretching vibration peaks of hydroxyl groups, and 1200 - 1030 cm -1 are the stretching vibration peaks of C-O. The C-O stretching vibration peak that appears in pullulan-grafted poly-(lactic acid-glycolic acid) at 1754 cm -1 , and the saturated carbon-hydrogen bond CH3 peak in the copolyester that appears at 3001 cm -1 indicate that the polyester type has been successfully introduced into the main chain of pullulan polysaccharide; Figure 3 As can also be seen, pullulan-grafted poly-(lactic acid-glycolic acid) has two weight loss processes. The first weight loss is at 120 - 280 °C, which is caused by the degradation of poly(lactic acid-glycolic acid) in the polymer; the second weight loss is at 280 - 480 °C, which is caused by the degradation of polysaccharide.

[0060] Example 1.1

[0061] The method of Example 1 is adopted, in which the molar ratio of stannous octoate to lactide is 0.06:100, the molar ratio of lactide to the hydroxyl groups of pullulan polysaccharide is 85:100, and the molar ratio of glycolide to lactide is 6:100.

[0062] Example 1.2

[0063] The method of Example 1 is adopted, in which the molar ratio of stannous octoate to lactide is 0.24:100, the molar ratio of lactide to the hydroxyl groups of pullulan polysaccharide is 60:100, and the molar ratio of glycolide to lactide is 10:100.

[0064] Example 1.3

[0065] Add to the Prussian polysaccharide , after heating and stirring for dissolution, lactide and glycolide are added. Under nitrogen protection, after mixing evenly, stannous octoate is added. At 80 °C, microwave reaction is carried out for 3 - 8 min. Among them, the molar ratio of stannous octoate to lactide is 0.15:100, the molar ratio of lactide to the hydroxyl group of pullulan is 75:100, and the molar ratio of glycolide to lactide is 5:100.

[0066] Blank nanoparticle aqueous dispersions were respectively prepared from the polymers obtained in the above examples, and their thermosensitive properties in water were measured by dynamic light scattering method. The results are as Figure 4 shown, Figure 4 A - 4C are the aqueous solutions of the nanoparticles prepared in Examples 1.1 - 1.3 respectively. When the system temperature reaches a critical value, the particle size of the nanoparticles will increase sharply. The critical value of this temperature is the lowest critical solution temperature ( ). Figure 4 It can be seen from Figure 4 that the of A - 4C are approximately 40, 38 and 32 °C.

[0067] Example 2 Preparation of drug - loaded nanoparticles

[0068] In order to achieve a better drug - controlled release effect, the inventor selected the pullulan - grafted poly-(lactic - glycolic acid) prepared in Example 1.1 as the drug self - assembly material;

[0069] A drug and pullulan - grafted poly-(lactic - glycolic acid) with a mass ratio of 1:(9 - 12) are dissolved in methanol, and heated under reduced pressure at 37 °C to obtain a composite film; the composite film and a 5% phosphate buffer solution are mixed and stirred in an ice - water bath for 10 - 20 min to obtain a suspension; the suspension is centrifuged, and the supernatant is filtered through a 0.8 - μm filter membrane to remove the unloaded raw materials, and a nanoparticle solution is prepared. The obtained nanoparticle solution is freeze - dried to obtain; among them, the drug includes 53 - 65%wt of bamboo leaf flavonoids, 23 - 42%wt of sugarcane polyphenols and 7 - 11% of phytosterols.

[0070] During the preparation of the drug - loaded nanoparticles, with the removal of methanol, the interaction between the drug and the hydrophobic end of poly-(lactic - glycolic acid) - modified pullulan gradually strengthens, and finally a composite film of the two is formed. When the aqueous phase is added, due to the strong interaction between the two that has been formed, the two together serve as the hydrophobic segment and are wrapped into the core of the nanoparticles, and the hydrophilic outer shell avoids the contact between the drug and the aqueous environment and reduces the precipitation of the drug. The lower critical solution temperature of poly-(lactic - glycolic acid) - modified pullulan in the aqueous phase is about 40 °C. Along with the increase of temperature, secondary aggregation will occur to form new aggregates with a larger volume, and when the temperature decreases again, the new aggregates will dissociate back into the original small aggregates;

[0071] Figure 1 This is the TEM image of the drug-loaded nanoparticles. As can be seen from the figure, the particle size of the drug-loaded nanoparticles is 90 - 100 nm, and its drug loading can reach 57.8 ± 0.5%, and the encapsulation efficiency reaches 81.6 ± 2.1%. The high encapsulation efficiency and drug loading can prolong the circulation time of the drug in the blood and improve the bioavailability of the drug.

[0072] Example 2.1

[0073] The method of Example 2 was adopted, wherein the mass ratio of the drug to pullulan-grafted poly-(lactic acid-glycolic acid) was 1:9; the drug included 53%wt of bamboo leaf flavonoids, 40%wt of sugarcane polyphenols, and 7% of phytosterols.

[0074] Example 2.2

[0075] The method of Example 2 was adopted, wherein the mass ratio of the drug to pullulan-grafted poly-(lactic acid-glycolic acid) was 1:10; the drug included 65%wt of bamboo leaf flavonoids, 24%wt of sugarcane polyphenols, and 11% of phytosterols.

[0076] Example 2.3

[0077] The method of Example 2 was adopted, wherein the mass ratio of the drug to pullulan-grafted poly-(lactic acid-glycolic acid) was 1:12; the drug included 58%wt of bamboo leaf flavonoids, 42%wt of sugarcane polyphenols, and 10% of phytosterols.

[0078] Comparative Example 1

[0079] Prepare a solution with a concentration of 30 mg / ml of bamboo leaf flavonoids and dilute it to the required concentration with cell culture medium by an appropriate dilution method, and filter it through a 0.2-μm filter membrane.

[0080] Perform membrane permeability tests on the drug-loaded nanoparticles prepared in the above examples and comparative examples:

[0081] Experimental animals: Male Wistar rats, grade, were raised in an environment with a constant temperature of 23 ± 2 °C and a light time from 8:00 to 20:00, fed freely, and given sterile water to drink. After one week of feeding, they were used for the experiment.

[0082] Prepare (Krebs-Ringer) solution: 7.8 g of sodium chloride, 0.22 g of magnesium chloride, 0.37 g of calcium chloride, 0.35 g of potassium chloride, 1.37 g of sodium bicarbonate, 0.32 g of sodium dihydrogen phosphate, 1.4 g of glucose. Place them in a 1000-ml volumetric flask, add distilled water to make up the volume, and adjust the pH to 7.40 with HCl solution to obtain liquid.

[0083] Experimental method: Take rats that have fasted for 12 hours and inject 20% urethane (10ml / kg) intraperitoneally. After the rats are anesthetized, fix them and maintain body temperature. Open the abdominal cavity at the midline of the abdomen, quickly separate the duodenum, and immerse it in 37℃ The intestinal sac was placed in the liquid and 95% O2 + 5% CO2 medical oxygen was continuously added to the nutrient solution. The surface of the small intestine was flushed with the nutrient solution to remove the mesentery and fat tissue on the serosal surface. The intestinal segment was turned over with the push rod in the syringe so that the mucosal layer was on the outside and the serosal layer was on the inside. The intestinal contents were cleaned and one end of the intestinal segment was ligated with surgical sutures and the other end was fixed on the outer tube of the 1ml syringe. The intestinal sac was placed in a 30ml syringe containing the corresponding preparation. The reaction tube of nutrient solution dilution was kept warm at 37°C in a water bath and stirred. 2 ml was injected into the intestinal sac. Nutrient solution was added, and medical oxygen was continuously introduced. After 2 hours, the intestinal sac fluid was drawn to determine the drug content, and the intestinal sac area was measured to calculate the amount of bamboo leaf flavonoids absorbed. The results are shown in Table 1.

[0084] Table 1

[0085]

[0086] It can be seen that drug-loaded nanoparticles can significantly improve the permeability of active drug ingredients, improve intestinal absorption, and increase bioavailability.

[0087] Example 3 Enteric Coated Preparation

[0088] Take the drug-loaded nanoparticles (prepared in Example 2.2), natto powder, water-insoluble dietary fiber, kudzu root powder and silicon dioxide, stir and mix them thoroughly to prepare inclusion powder with a particle size of 0.6-1.5 mm; the inclusion powder is coated with an enteric coating layer in a fluidized bed, and the parameters of the fluidized bed are controlled as follows: the air volume is 270-330m 3 / h, the drug application rate is 6g-15g / min, the air inlet temperature is 50-60℃, the material temperature is 32-35℃, and the atomization pressure is 0.1MPa-0.3MPa; the suspension stirring speed is 500-750rpm, and the stirring time is 25-35min; the mass ratio of enteric coating to inclusion powder is (0.15-0.25):1 to obtain an enteric preparation; the enteric coating comprises 65 parts of polyacrylic acid resin, 11 parts of propylene glycol, and 24 parts of 95% ethanol aqueous solution; the release amount of the enteric preparation in artificial gastric juice within 4 hours is less than 10%, and the release amount in artificial intestinal juice within 4 hours is greater than 90%.

[0089] Example 3.1

[0090] The method of Example 3 is adopted, including 280 parts of drug-loaded nanoparticles, 4 parts of natto powder, 200 parts of water-insoluble dietary fiber, and 250 parts of kudzu root powder; the mass ratio of enteric coating to inclusion powder is 0.15:1.

[0091] Example 3.2

[0092] The method of Example 3 is adopted, including 290 parts of drug-loaded nanoparticles, 5 parts of natto powder, 150 parts of water-insoluble dietary fiber, and 225 parts of kudzu root powder; the mass ratio of enteric coating to inclusion powder is 0.25:1.

[0093] Example 3.3

[0094] The method of Example 3 is adopted, including 310 parts of drug-loaded nanoparticles, 4.5 parts of natto powder, 180 parts of water-insoluble dietary fiber, and 280 parts of kudzu root powder; the mass ratio of enteric coating to inclusion powder is 0.18:1.

[0095] Example 3.4

[0096] The method of Example 3 is adopted, including 300 parts of drug-loaded nanoparticles, 5 parts of natto powder, 180 parts of water-insoluble dietary fiber, and 300 parts of kudzu root powder; the mass ratio of enteric coating to inclusion powder is 0.20:1.

[0097] Example 3.5

[0098] The method of Example 3 is adopted, including 300 parts of drug-loaded nanoparticles, 4.5 parts of natto powder, 200 parts of water-insoluble dietary fiber, and 260 parts of kudzu root powder; the mass ratio of enteric coating to inclusion powder is 0.25:1.

[0099] Example 3.6

[0100] The method of Example 3 is adopted, including 310 parts of drug-loaded nanoparticles, 5 parts of natto powder, 180

[0101] parts of water-insoluble dietary fiber, and 250 parts of kudzu root powder; the mass ratio of enteric coating to inclusion powder is 0.18:1.

[0102] Comparative Example 2

[0103] Same as Example 3.1, except that the drug loading amount of bambusae flavones in the drug-loaded nanoparticles is 0;

[0104] Comparative Example 3

[0105] Same as Example 3.2, except that the drug loading amount of sugarcane polyphenols in the drug-loaded nanoparticles is 0;

[0106] Comparative Example 4

[0107] Same as Example 3.3, except that the drug loading of phytosterol in the drug-loaded nanoparticles is 0;

[0108] Comparative Example 5

[0109] Same as Example 3.4, except that natto powder is not added;

[0110] Comparative Example 6

[0111] Same as Example 3.5, except that insoluble dietary fiber is not added;

[0112] Comparative Example 7

[0113] Same as Example 3.6, except that kudzu root powder is not added.

[0114] The hypolipidemic effect of the enteric-coated preparations prepared in the above examples and comparative examples was tested:

[0115] (1)Hypolipidemic effect test

[0116] Preparation of high-fat emulsion: Take Lard is heated and melted, and rapeseed oil and cholesterol are added and dissolved, then sodium cholate and propylthiouracil are added and stirred and mixed , then Tween is added, 20 mL of propylene glycol and 50 mL of distilled water are added, stirred and emulsified at 10000 r / min for 5 min, cooled to room temperature, and distilled water is continuously added to 100 mL, and mixed for 10 min to obtain a fat emulsion.

[0117] Establishment of hyperlipidemia model: Each group of rats was intragastrically administered high-fat emulsion on an empty stomach at 8 am every day according to a dose of 10 mL / (kg·d), and freely fed ordinary feed and freely drank water at other times. The rats were weighed once a week and continuously gavaged for 4 weeks.

[0118] Grouping and dosing regimen: The rats were divided into a normal group, a model group, an example group, and a comparative example group, with 3 rats in each group. The normal group and the model group were given an equal amount of normal saline, and the other groups were all given high-fat emulsion. The normal group. The experimental group was given drug intervention treatment while modeling. The example group and the comparative example group were given the drug-loaded nanoparticles prepared in Examples 3.1 - 3.6 or Comparative Examples 2 - 7 once at 16:00 every afternoon according to 10 g / (kg·d) per day.

[0119] Index detection: The test period was 4 weeks. After the last gavage, the mice were fasted for 5 hours, and the animals in each group were treated. The rats were weighed, anesthetized, and blood was collected from the heart. The blood was anticoagulated with heparin, centrifuged, and plasma was separated. The concentrations of total cholesterol ( ), triglyceride ( ), high-density lipoprotein cholesterol ( ), and low-density lipoprotein cholesterol ( ), in the plasma of rats were detected by using a kit method, and the ratio of was calculated. The livers of the mice were separated, and the contents of catalase ( ), total superoxide dismutase ( ), and glutathione peroxidase ( ) in the liver tissues were determined by using a kit method. The protein concentration was determined by using a assay kit. The results are shown in Table 2.

[0120] Table 2

[0121]

[0122] Hyperlipidemia is a common chronic disease, which is likely to cause cardiovascular and cerebrovascular diseases such as atherosclerosis. It is usually due to excessive intake of lipids or disorder of lipid metabolism in the body. The specific manifestation is that the , and in the serum are too high, and is too low. and are important indicators for evaluating cholesterol metabolism in the body. The function of is to reverse transport cholesterol from tissues outside the liver back to the liver. As can be seen from the above table, the enteric-coated preparation prepared in the embodiment of the present invention can significantly reduce the contents of , , in the serum, and increase the content of . Compared with the embodiment, the contents of , and in each comparative example are increased, and the content of is decreased. It can be seen that each component has the effect of assisting in reducing blood lipids, and bamboo leaf flavonoids, sugarcane polyphenols and phytosterols have a significant effect on regulating blood lipids. The addition of the three has a synergistic effect, which can promote the decrease of total cholesterol and low-density lipoprotein cholesterol in patients with hyperlipidemia, inhibit cholesterol synthesis, increase the content of , and reduce the contents of , , playing a very good role in regulating blood lipids.

[0123] (2) In vitro dissolution test:

[0124] The dissolution degrees of the enteric-coated preparations of the examples and comparative examples at different times in artificial gastric juice and artificial intestinal juice were investigated, and the test results are shown in Table 3. Artificial gastric juice and artificial intestinal juice were prepared according to the Chinese Pharmacopoeia 2005 edition. The specific preparation method of artificial gastric juice: Take 9 ml of 0.365 g / ml hydrochloric acid, dilute it with distilled water to 1 L, and add pepsin to make its mass fraction 1%, then it is obtained; the specific preparation method of artificial intestinal juice: Take 6.8 g of KH2PO4, add 500 ml of water to dissolve it, adjust the pH to 6.8 with 0.1 mol NaOH solution; another take 10 g of pancreatin, dissolve it with an appropriate amount of water; mix the two solutions and dilute with water to 1000 ml, then it is obtained.

[0125] Table 3

[0126]

[0127] It can be seen that the enteric-coated preparation of the present invention is basically not released in artificial gastric juice, and shows the characteristics of slow release in artificial simulated intestinal juice, the cumulative release amount increases slowly, and the cumulative release time can reach more than 4 hours.

[0128] Finally, it should be noted that the above description is only the preferred embodiment of the present invention. Those skilled in the art can make various similar representations under the inspiration of the present invention without violating the purpose and claims of the present invention. Such transformations all fall within the protection scope of the present invention.

Claims

1. A composition for regulating blood lipid levels, characterized in that It comprises raw materials in the following parts by mass: 280 - 310 parts of drug-loaded nanoparticles, 4 - 5 parts of natto powder, 150 - 200 parts of water-insoluble dietary fiber, and 225 - 300 parts of kudzu root powder; wherein, the particle size of the drug-loaded nanoparticles is 90 - 100 nm, and the drug loading of the drug-loaded nanoparticles is 57.8 ± 0.5%; the drug comprises 53 - 65%wt of bamboo leaf flavonoids, 23 - 42%wt of sugarcane polyphenols, and 7 - 11% of phytosterols; the drug-loaded nanoparticles are prepared by the following method: dissolving a drug and pullulan-grafted poly-(lactic acid-glycolic acid) with a mass ratio of 1:(9 - 12) in methanol, heating under reduced pressure at 37°C to obtain a composite film; mixing the composite film and a hydration solution, stirring and hydrating in an ice-water bath for 10 - 20 min to obtain a suspension; centrifuging and separating the suspension, taking the supernatant and filtering, filtering with a microporous membrane to remove the unloaded raw materials to prepare a nanoparticle solution, and freeze-drying the obtained nanoparticle solution to obtain the product; wherein, the hydration solution is a phosphate buffer solution with a mass fraction of 3 - 10%.

2. The composition for regulating blood lipid level according to claim 1, wherein: The hydration solution is a phosphate buffer solution with a mass fraction of 5 - 8%.

3. A composition for regulating blood lipid levels according to claim 1, characterized in that: The hydration solution is a phosphate buffer solution with a mass fraction of 5.5%.

4. A composition for regulating blood lipid levels according to claim 1, characterized in that, The pullulan-grafted poly-(lactic acid-glycolic acid) is prepared by the following method: adding DMSO to pullulan polysaccharide, heating and stirring to dissolve, then adding lactide and glycolide, under nitrogen protection, mixing evenly and then adding stannous octoate, reacting by microwave at 80°C for 3 - 8 min, wherein the molar ratio of stannous octoate to lactide is (0.06 - 0.24):100, the molar ratio of lactide to the hydroxyl group of pullulan polysaccharide is (60 - 85):100, and the molar ratio of glycolide to lactide = (5 - 10):

100.

5. A composition for regulating blood lipid levels according to any one of claims 1-4, characterized in that: The water-insoluble dietary fiber is selected from one or more of oat fiber, bamboo shoot fiber, wheat fiber, corn fiber, soybean fiber, and pea fiber.

6. An enteric-coated preparation, characterized in that: It comprises the composition according to any one of claims 1 - 5 and an enteric coating; the release amount of the enteric preparation in artificial gastric juice within 4 hours < 10%, and the release amount in artificial intestinal juice within 4 hours > 90%.

7. An enteric preparation according to claim 6, characterized in that: Calculated by 100 parts by mass, the enteric coating comprises 60 - 75 parts of film-forming material, 8 - 12 parts of plasticizer, and the balance is solvent.

8. An enteric preparation according to claim 6, characterized in that: Calculated by 100 parts by mass, the film-forming material is 65 - 70 parts, the plasticizer is 10 - 12 parts, and the balance is solvent.

9. An enteric preparation according to claim 6, characterized in that: Calculated by 100 parts by mass, the film-forming material is 68 parts, the plasticizer is 11 parts, and the balance is solvent.

10. An enteric preparation according to any one of claims 6 - 9, characterized in that: The film-forming material is selected from one or more of polyacrylic resin, hydroxypropylmethylcellulose acetate succinate, cellulose acetate phthalate, ethyl cellulose, polyvinyl alcohol phthalate, enteric Opadry, carboxymethylethyl cellulose, polyvinyl acetate phthalate, and diketopiperazine polymer; the plasticizer is selected from one or more of polyethylene glycol, propylene glycol, triethyl citrate, diethyl phthalate, tributyl citrate, and glycerol monostearate and distearate.

11. A method for preparing an enteric-coated preparation according to any one of claims 6-10, characterized in that It includes the following steps: fully stirring and mixing drug-loaded nanoparticles, natto powder, water-insoluble dietary fiber, kudzu root powder and an adsorbent to prepare an inclusion powder with a particle size of 0.6 - 1.5 mm; performing enteric coating on the inclusion powder in a fluidized bed; the adsorbent is one or more of silica, cyclodextrin, lactose, mannitol, calcium hydrogen phosphate, calcium sulfate, microcrystalline cellulose and acrylic resin.

12. The preparation method according to claim 11, characterized in that: The parameters of the fluidized bed are as follows: the air intake is controlled at 270 - 330 m 3 / h, the rate of adding medicine is 6 g - 15 g / min, the air inlet temperature is 50 - 60 °C, the material temperature is 32 - 35 °C, and the atomization pressure is 0.1 MPa - 0.3 MPa; the suspension stirring speed is 500 - 750 rpm, and the stirring time is 25 - 35 min.

13. The preparation method according to claim 11, characterized in that: The mass ratio of the enteric coating to the inclusion powder is (0.15 - 0.25):

1.

14. The preparation method according to claim 11, characterized in that: The mass ratio of the enteric coating to the inclusion powder is (0.18 - 0.22):

1.

15. The preparation method according to claim 11, wherein: The mass ratio of the enteric coating to the inclusion powder is 0.2:1.

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