Preparation process of Dendrobium officinale composite granules for clearing liver and improving eyesight

Abstract

The invention relates to a preparation process of a liver-clearing and eyesight-improving dendrobium officinale composite particle. Firstly, Chinese medicine particles are extracted and separated to separate insoluble substances, and the insoluble substances are first modified with amino acid derivatives to improve solubility and bioavailability, and improve the efficacy of liver-clearing and eyesight-improving. Then, nano suspension is prepared, and a suitable surfactant is selected to further increase the compatibility of the insoluble components. Finally, a suitable wetting agent is selected in the process of spray granulation to improve the stability and dispersibility of the drug particles. The compound particles have a unique taste and have the effects of clearing the liver, improving eyesight, detoxifying, refreshing the brain, stimulating appetite, and the like. Long-term drinking can protect eyes, protect vision, and prevent myopia.

Description

Technical Field

[0001] The invention relates to a method for preparing a dendrobium officinale composite, in particular to a process for preparing liver-clearing and eye-improving dendrobium officinale composite particles. Background Art

[0002] Eyes are human visual organs, but modern people often need to face electronic devices such as computer screens and mobile phone screens. Long-term use of eyes can easily lead to problems such as eye fatigue, dryness, and decreased vision. Granular preparations refer to granular and block preparations made from medicinal material extracts and suitable auxiliary materials or medicinal material fine powders. They are easy to carry and use and are deeply loved by consumers. There are many varieties of granular granules in the market. Most granules are only used as energy drinks with general health effects. At present, there is no dendrobium composite granule that can nourish the liver and improve eyesight and has a good taste. The present invention improves the process of the formula of Dendrobium officinale luminous pills, which is used to prepare a granular preparation with a unique taste, which has the effects of clearing the liver, improving eyesight, detoxifying, refreshing the brain, and appetizing. Long-term drinking can protect the eyes, protect eyesight, and prevent myopia.

[0003] Recent studies have shown that more than 40% of the main active ingredients of Chinese medicine in my country are poorly soluble ingredients, and such ingredients have a wide range of pharmacological activities, such as anti-tumor, anti-inflammatory, antiviral, anti-oxidant, anti-thrombotic, etc. However, due to low solubility, poor gastrointestinal absorption, blood drug concentrations are prone to peaks and valleys, and oral bioavailability is low, which seriously limits the clinical efficacy. In order to improve the clinical therapeutic effect of the poorly soluble active ingredients of Chinese medicine, it is found that introducing amino acids into drugs as carriers can not only improve the problems of poor solubility, but also improve the corresponding pharmacological activity. Therefore, the present invention is directed to the introduction of amino acids to study how to improve the liver-clearing and eye-improving effects of the composite preparation at multiple targets.

[0004] There are many methods to solve the problems of poor water solubility and low bioavailability of drugs, such as co-solvent solubilization, cyclodextrin inclusion, micronization technology and preparation of intravenous emulsions, but these commonly used technologies have certain limitations. For example, co-solvents require drugs to have specific physical and chemical properties (such as being soluble in certain organic solvents), and there are problems such as organic solvent toxicity and side effects, and drug precipitation during compatibility; cyclodextrin inclusion has special requirements for the size of drug molecules; micronization method has no obvious effect on increasing bioavailability; emulsions require drugs to have a higher solubility in the oil phase. Subsequent studies have found that nanosuspensions can better solve the above problems. Using the stabilizing effect of surfactants, the crushed Chinese medicine particles are dispersed in water, and a stable nano-colloidal dispersion is formed through technical control. This method is simple, fast to prepare, with less auxiliary additions and reduced toxic side effects.

[0005] Alkanolamide compounds have strong hydrolysis resistance, low toxicity, good biodegradability, and no skin irritation due to the presence of amide bonds in their molecules. They are excellent nonionic surfactants and are also intermediates for further synthesizing other surfactants. Therefore, the present invention preferentially selects the most suitable surfactant from the alkanolamide compounds and applies it to the nanosuspension.

[0006] Nanosuspensions are thermodynamically unstable systems with poor physical stability. Crystal growth, particle aggregation, and sedimentation often occur during storage. Inconvenience in carrying and taking is a major disadvantage of nanosuspension liquid preparations, which seriously restricts the application and promotion of nanosuspensions. Nanosuspensions are dehydrated and solidified into drug powders by vacuum drying, spray drying, freeze drying, etc., or further processed into various dosage forms according to different administration routes, such as tablets, capsules, aerosols, and dry powder suspensions. On the one hand, the problem of thermodynamic instability of the nanosuspension system can be solved to improve the storage stability of the preparation; on the other hand, it is convenient to use and carry. After oral administration of the drug nanosuspension solidified powder, it can be redispersed into a nanoparticle state under the wetting effect of the gastrointestinal liquid in the body to obtain the same effect as the initial nanosuspension liquid preparation, so as to improve the solubility and bioavailability of the drug. Therefore, research on the solidification and redispersion of nanosuspensions will open up new breakthroughs for the industrialization of the preparation.

[0007] Fluidized bed spray drying technology is a drying method that combines spray drying technology with fluidization technology. It is to place tiny particles or pellets such as starch and sucrose in a fluidized bed in a boiling state, and the drug solution or suspension is sprayed out through an atomizer, first undergoing heat exchange with the hot air flow, and then sprayed onto the particle mother core or blank pellet in the fluidized bed at the bottom of the tower, and dried on the surface to make it grow continuously, and finally become dry particles or micro-pellets. Fluidized bed spray drying has the advantages of spray drying technology and can achieve one-step granulation molding.

[0008] Different composition of ingredients has a great influence on the mechanical properties of pills during the drying process. Pills with more polysaccharide components are compact, have rough surfaces and shrinkage after drying, and have poor disintegration; pills with more fiber components have a rough surface, poor pilling, and good disintegration; pills with more starch have better formability, smooth surface, and can dissolve quickly after absorbing water. The different composition of pills has a great influence on their texture properties. The main texture properties test items of pills reported so far include hardness, cohesion, elasticity, etc. Traditional Chinese medicine pills have a complex amorphous polymer composition system. Improper control of the drying process can easily lead to uneven drying of pills and quality problems such as cracked pills. By selecting the optimal fluidized bed spray technology and preparing a suitable wetting agent, it is used to solve problems such as unevenness of granular preparations. Summary of the invention

[0009] In order to solve the above technical problems, the present invention provides a solution.

[0010] The specific steps are:

[0011] (1) Extraction and separation: Wash, crush and dry 60 parts of Dendrobium officinale, 80 parts of Ginseng, 20 parts of Chinese Yam, 60 parts of Poria, 20 parts of Licorice, 20 parts of Cistanche, 40 parts of Lycium barbarum, 20 parts of Cuscuta, 60 parts of Rehmannia, 30 parts of Schisandra, 80 parts of Asparagus, 50 parts of Ophiopogon, 25 parts of Apricot, 30 parts of Saposhnikovia, 30 parts of Chuanxiong, 20 parts of Citrus aurantium, 20 parts of Coptis, 35 parts of Achyranthes, 25 parts of Chrysanthemum, 2 0 parts of Tribulus terrestris, 20 parts of Amaranthus seeds, 35 parts of Cassia seeds, 50 parts of buffalo horn concentrated powder, and 220-260 parts of antelope horn are crushed through a 50-60 mesh sieve; the mixed powder is put into an extraction kettle, and supercritical CO2 extraction is performed, the critical temperature is 30-35°C, the critical pressure is 7-9Mpa, the extraction time is 4-6h, the entrainer is anhydrous ethanol, and the entrainer dosage is 20%, to obtain an extract and a filter residue with insoluble substances;

[0012] (2) The extract is separated and purified using d101b weakly polar macroporous resin, and then the extract is dialyzed in running water for 24 hours at a dialysis temperature of 30°C, with the water changed every 4 hours; then the extract is vacuum concentrated at 45-55°C to 1 / 10-1 / 3 of the original volume, 1-3 times of ethanol is added to the concentrate for precipitation, separated using Sephadex G75 gel, and finally vacuum freeze-dried at -70-78°C for 12-24 hours to obtain soluble particles;

[0013] (3) Amino acid modification: 2-4 parts of an amino acid derivative were dissolved in 40 parts of a THF solution, cooled in an ice bath and stirred for 30 min, 2-4 parts of the above-mentioned insoluble substance in the filter residue were added, and the reaction was continued with stirring at room temperature for 2 h, filtered, and the filtrate was concentrated and purified by silica gel column chromatography, wherein the volume ratio of ethyl acetate to petroleum ether was 1:6, and it was used as an eluent for purification to obtain solid particles;

[0014] The amino acid derivative is Boc-L-cyclohexylalanine.

[0015] (4) Nanosuspension: Weigh 10-20 parts of the above solids, add them into distilled water containing the surface suspending agent, stir and mix thoroughly, and ultrasonicate at 8000 r / min for 10 min to obtain a coarse suspension. Pass the coarse suspension through a high-pressure homogenizer at a pressure of 1000 bar for 15 cycles and set aside;

[0016] The distilled water containing the surface suspension is: 10-15 parts of poloxamer 188, 8-12 parts of sodium dodecyl hydrogen sulfate, and 5-8 parts of a stabilizing regulator are added to 200 parts of distilled water and stirred for decomposition for 10 minutes; wherein the stabilizing regulator is monoethanolamide of dodecenoic acid.

[0017] (5) Granulation: The above suspension and soluble granules are mixed in a ratio of 4:6, and stirred in a rapid stirring pot for 10-15 minutes. The wet granules are quickly placed in a centrifugal coating granulator, and the main engine speed is adjusted to 100-120 r / min. The granules are spheronized for 30 minutes. During this period, 10% of the wetting agent is sprayed every 10 minutes. After that, the mixture is dried for 5 hours at a hot air temperature of 50-60°C and a blast flow rate of 20 L / min to obtain the compound Dendrobium officinale granule preparation for clearing the liver and improving eyesight.

[0018] The preparation of the wetting agent is as follows: prepare 3-5 parts of PVP and 20-30 parts of ethanol, heat to 40°C and stir for 5 minutes, add 0.5-1 part of a wetting regulator, and mix; wherein the wetting regulator is adenosylmethionine p-toluenesulfonate.

[0019] The beneficial effects of the present invention are:

[0020] 1. The present invention relates to a preparation process of a liver-clearing and eyesight-improving Dendrobium officinale composite particle. First, Chinese medicine particles are extracted and separated to separate insoluble substances. The insoluble substances are first modified with amino acid derivatives to improve solubility and bioavailability, and improve the efficacy of liver-clearing and eyesight-improving. Then, a nano suspension is prepared, and a suitable surfactant is selected to further increase the compatibility of the insoluble components. Finally, a suitable wetting agent is selected in the spray granulation process to improve the stability and dispersibility of the drug particles. The compound particles have a unique taste and have the effects of clearing the liver, improving eyesight, detoxifying, refreshing the brain, and stimulating appetite. Long-term drinking can protect the eyes, protect vision, and prevent myopia.

[0021] 2. The study found that as the volume of the hydrocarbon group introduced into the alanine derivative increases, the antioxidant activity also increases. Among them, the antioxidant activity of the cyclohexylalanine derivative introduced is significantly better than that of other hydrocarbon alanine derivatives. The cyclohexylalanine derivative has a better stable structure and is easy to react with other groups to generate new molecules. The present invention uses Boc-L-cyclohexylalanine to carry out amino acid derivatization of the insoluble components of traditional Chinese medicine, which significantly improves the antioxidant activity of the drug, increases the water solubility of the insoluble components, improves its oral bioavailability and improves the effect of clearing the liver and improving eyesight.

[0022] 3. The introduction of amino acid derivatives and the synthesis of new substances can not only down-regulate the expression of c-Fos and c-Jun in the injured retina, up-regulate the expression of Bcl2, enhance the role of antagonizing photoreceptor cell apoptosis induced by light damage, reduce the mortality rate of photoreceptor cells, but also protect the visual system at multiple targets, maintain visual clarity, improve light adaptation, and better maintain eye health.

[0023] 4. The monoethanolamide of dodecenoic acid has the structural characteristics of alkanolamide nonionic surfactants, has strong hydrolysis resistance, low toxicity, good biodegradability, and does not irritate the skin. It is an excellent nonionic surfactant with a wider applicable pH range than general ionic surfactants. It can spontaneously form micelles in aqueous solution to solubilize insoluble components. The monoethanolamide of dodecenoic acid can adhere to the surface of Dendrobium officinale particles to form a three-dimensional spatial repulsive structure, prevent the particles from aggregating with each other, and control the particle size of the raw material. The highly uniform particle size increases the physical stability of the drug, avoids Ostwald aging, and increases the dissolution rate of the drug.

[0024] 5. In addition, if other acidic surfactants are selected, the pH value of the system is close to the acidic pKa, resulting in a high degree of protonation of the poorly soluble substances, making it difficult to provide good repulsion stability. Compared with other acidic substances, the long-chain olefinic acid in the monoethanolamide structure of dodecenoic acid has an average particle size closer to that of poorly soluble substances and a higher pKa, and has a better spatial effect. It can form a network structure through intermolecular forces, inhibit the aggregation and sedimentation of nanoparticles, and make the Chinese medicine particles in the solution easier to disperse, thereby improving its compatibility with other materials.

[0025] 6. Adenosylmethionine p-toluenesulfonate is selected as a wetting regulator, which has high stability under high temperature and common humidity conditions. Compared with similar regulators, it is not easy to decompose or lose activity, and it has strong buffering properties, which can enhance the stability of the active ingredients in the Dendrobium Officinale Granules. Through the spraying process during the stirring process, the droplets first wet the powder they come into contact with and aggregate around it to form a particle core. Adenosylmethionine p-toluenesulfonate can absorb and store moisture in the surrounding environment to a certain extent, release moisture when the humidity is high, and absorb moisture when the humidity is low, thereby maintaining the humidity balance of the extract material and ensuring the stability of the granule product.

[0026] 7. Adenosylmethionine toluenesulfonate as a polypeptide molecule can also be used to regulate metabolism and promote absorption. The toluenesulfonic acid-specific functional group in adenosylmethionine toluenesulfonate has good dispersibility, which can improve the phenomenon of moisture absorption, softening, agglomeration and adhesion between cellulose small molecules in traditional Chinese medicine granules, improve agglomeration between mixtures and adhesion between particles, ensure the product quality of powders, and with the cooperation of other excipients, greatly improve the distribution of particle size and the fluidity of powders. DETAILED DESCRIPTION

[0027] The present invention will be further described in detail below in conjunction with the embodiments.

[0028] Unless otherwise specified, the raw materials or chemical reagents used in the embodiments of the present invention are obtained through conventional commercial channels.

[0029] Example 1

[0030] (1) Extraction and separation: Wash, crush and dry 60 parts of Dendrobium officinale, 80 parts of Ginseng, 20 parts of Chinese Yam, 60 parts of Poria, 20 parts of Licorice, 20 parts of Cistanche, 40 parts of Lycium barbarum, 20 parts of Cuscuta, 60 parts of Rehmannia root, 30 parts of Schisandra chinensis, 80 parts of Asparagus cochinchinensis, 50 parts of Ophiopogon japonicus, 25 parts of Apricot kernel, 30 parts of Saposhnikovia divaricata, 30 parts of Chuanxiong, 20 parts of Citrus aurantium, 20 parts of Coptis chinensis, 35 parts of Achyranthes bidentata, 25 parts of chrysanthemum, 20 parts of tribulus, 20 parts of cyperus, 35 parts of cassia seed, 50 parts of buffalo horn concentrated powder, 240 parts of antelope horn, are crushed and passed through a 50-mesh sieve; the mixed powder is put into an extraction kettle, and supercritical CO2 extraction is performed, the critical temperature is 32°C, the critical pressure is 8Mpa, the extraction time is 5h, the entrainer is anhydrous ethanol, and the entrainer dosage is 20%, to obtain an extract and a filter residue with insoluble substances;

[0031] (2) The extract was separated and purified using d101b weakly polar macroporous resin, and then dialyzed in running water for 24 h at a dialysis temperature of 30°C, with the water changed every 4 h; then vacuum concentrated to 1 / 6 of the original volume at 50°C, 2 times ethanol was added to the concentrate for precipitation, separated using Sephadex G75 gel, and finally freeze-dried at -74°C in a vacuum for 18 h to obtain soluble particles;

[0032] (3) Amino acid modification: 3 parts of Boc-L-cyclohexylalanine were dissolved in 40 parts of THF solution, cooled in an ice bath and stirred for 30 min, 3 parts of the above-mentioned insoluble substance in the filter residue were added, and the reaction was continued with stirring at room temperature for 2 h, filtered, and the filtrate was concentrated and purified by silica gel column chromatography, wherein the volume ratio of ethyl acetate to petroleum ether was 1:6, and it was used as the eluent for purification to obtain solid particles;

[0033] (4) Nanosuspension: 12 parts of poloxamer 188, 10 parts of sodium dodecyl sulfate, and 7 parts of monoethanolamide of dodecenoic acid were added to 200 parts of distilled water, and the mixture was stirred and decomposed for 10 minutes to obtain distilled water containing a surface suspending agent; 15 parts of the solid particles obtained in the previous step were weighed, added to the distilled water containing the surface suspending agent, and the mixture was thoroughly stirred and mixed. After ultrasonic treatment at 8000 r / min for 10 minutes, a coarse suspension was obtained. The coarse suspension was passed through a high-pressure homogenizer at a pressure of 1000 bar for 15 cycles and kept ready for use;

[0034] (5) The wetting agent is prepared as follows: 4 parts of PVP and 25 parts of ethanol are prepared, heated to 40° C. and stirred for 5 min, and 0.8 parts of adenosylmethionine p-toluenesulfonate are added and mixed;

[0035] (6) Granulation: The above suspension and soluble granules are mixed in a ratio of 4:6, and stirred in a rapid stirring pot for 12 min. The wet granules are quickly placed in a centrifugal coating granulator, and the main engine speed is adjusted to 110 r / min. The granules are spheronized for 30 min. During this period, 10% of the wetting agent is sprayed every 10 min. After that, the mixture is dried at a hot air temperature of 55°C and a blast flow rate of 20 L / min for 5 h. The compound Dendrobium officinale granule preparation for clearing the liver and improving eyesight is obtained.

[0036] Example 2

[0037] (1) Extraction and separation: Wash, crush and dry 60 parts of Dendrobium officinale, 80 parts of Ginseng, 20 parts of Chinese Yam, 60 parts of Poria, 20 parts of Licorice, 20 parts of Cistanche, 40 parts of Lycium barbarum, 20 parts of Cuscuta, 60 parts of Rehmannia root, 30 parts of Schisandra chinensis, 80 parts of Asparagus cochinchinensis, 50 parts of Ophiopogon japonicus, 25 parts of Apricot kernel, 30 parts of Saposhnikovia divaricata, 30 parts of Chuanxiong, 20 parts of Citrus aurantium, 20 parts of Coptis chinensis, 35 parts of Achyranthes bidentata, 25 parts of chrysanthemum, 20 parts of tribulus, 20 parts of cyperus, 35 parts of cassia seed, 50 parts of buffalo horn concentrated powder, 220 parts of antelope horn, are crushed and passed through a 50-mesh sieve; the mixed powder is put into an extraction kettle, and supercritical CO2 extraction is performed, the critical temperature is 35°C, the critical pressure is 7Mpa, the extraction time is 6h, the entrainer is anhydrous ethanol, and the entrainer dosage is 20%, to obtain an extract and a filter residue with insoluble substances;

[0038] (2) The extract was separated and purified using d101b weakly polar macroporous resin, and then dialyzed in running water for 24 h at a dialysis temperature of 30°C, with the water changed every 4 h; then vacuum concentrated at 55°C to 1 / 10 of the original volume, 3 times ethanol was added to the concentrate for precipitation, separated using Sephadex G75 gel, and finally freeze-dried at -70°C for 24 h to obtain soluble particles;

[0039] (3) Amino acid modification: 2 parts of Boc-L-cyclohexylalanine were dissolved in 40 parts of THF solution, cooled in an ice bath and stirred for 30 min, 4 parts of the above-mentioned insoluble substance in the filter residue were added, and the reaction was continued with stirring at room temperature for 2 h, filtered, and the filtrate was concentrated and purified by silica gel column chromatography, wherein the volume ratio of ethyl acetate to petroleum ether was 1:6, and it was used as the eluent for purification to obtain solid particles;

[0040] (4) Nanosuspension: 10 parts of poloxamer 188, 12 parts of sodium dodecyl sulfate, and 5 parts of monoethanolamide of dodecenoic acid were added to 200 parts of distilled water, and the mixture was stirred and decomposed for 10 minutes to obtain distilled water containing a surface suspending agent; 20 parts of the solid particles obtained in the previous step were weighed, added to the distilled water containing the surface suspending agent, and the mixture was thoroughly stirred and mixed. After ultrasonication at 8000 r / min for 10 minutes, a coarse suspension was obtained. The coarse suspension was passed through a high-pressure homogenizer at a pressure of 1000 bar for 15 cycles and kept ready for use;

[0041] (5) The wetting agent is prepared as follows: 5 parts of PVP and 20 parts of ethanol, heated to 40° C. and stirred for 5 min, and 0.5 parts of adenosylmethionine p-toluenesulfonate are added and mixed;

[0042] (6) Granulation: The above suspension and soluble granules are mixed in a ratio of 4:6, and stirred in a rapid stirring pot for 15 minutes. The wet granules are quickly placed in a centrifugal coating granulator, and the main engine speed is adjusted to 100 r / min. The granules are spheronized for 30 minutes. During this period, 10% of the wetting agent is sprayed every 10 minutes. After that, the mixture is dried at a hot air temperature of 60°C and a blast flow rate of 20 L / min for 5 hours to obtain the compound Dendrobium officinale granule preparation for clearing the liver and improving eyesight.

[0043] Example 3

[0044] (1) Extraction and separation: Wash, crush and dry 60 parts of Dendrobium officinale, 80 parts of Ginseng, 20 parts of Chinese Yam, 60 parts of Poria, 20 parts of Licorice, 20 parts of Cistanche, 40 parts of Lycium barbarum, 20 parts of Cuscuta, 60 parts of Rehmannia root, 30 parts of Schisandra chinensis, 80 parts of Asparagus cochinchinensis, 50 parts of Ophiopogon japonicus, 25 parts of Apricot kernel, 30 parts of Saposhnikovia divaricata, 30 parts of Chuanxiong, 20 parts of Citrus aurantium, 20 parts of Coptis chinensis, 35 parts of Achyranthes bidentata, 25 parts of chrysanthemum, 20 parts of tribulus, 20 parts of cyperus, 35 parts of cassia seed, 50 parts of buffalo horn concentrated powder, 260 parts of antelope horn, are crushed and passed through a 60-mesh sieve; the mixed powder is put into an extraction kettle, and supercritical CO2 extraction is performed, the critical temperature is 30°C, the critical pressure is 9Mpa, the extraction time is 4h, the entrainer is anhydrous ethanol, and the entrainer dosage is 20%, to obtain an extract and a filter residue with insoluble substances;

[0045] (2) The extract was separated and purified using d101b weakly polar macroporous resin, and then dialyzed in running water for 24 h at a dialysis temperature of 30°C, with the water changed every 4 h; then vacuum concentrated to 1 / 3 of the original volume at 45°C, 1-fold ethanol was added to the concentrate for precipitation, separated using Sephadex G75 gel, and finally freeze-dried at -78°C for 12 h to obtain soluble particles;

[0046] (3) Amino acid modification: 4 parts of Boc-L-cyclohexylalanine were dissolved in 40 parts of THF solution, cooled in an ice bath and stirred for 30 min, 2 parts of the above-mentioned insoluble substance in the filter residue were added, and the reaction was continued with stirring at room temperature for 2 h, filtered, and the filtrate was concentrated and purified by silica gel column chromatography, wherein the volume ratio of ethyl acetate to petroleum ether was 1:6, which was used as the eluent for purification to obtain solid particles;

[0047] (4) Nanosuspension: 15 parts of poloxamer 188, 8 parts of sodium dodecyl hydrogen sulfate, and 8 parts of monoethanolamide of dodecenoic acid were added to 200 parts of distilled water, and stirred for 10 minutes to obtain distilled water containing a surface suspending agent; 10 parts of the solid particles obtained in the previous step were weighed, added to the distilled water containing the surface suspending agent, and stirred thoroughly to mix, and then subjected to ultrasonication at 8000 r / min for 10 minutes to obtain a coarse suspension, and the coarse suspension was passed through a high-pressure homogenizer at a pressure of 1000 bar for 15 cycles for standby use;

[0048] (5) The wetting agent is prepared as follows: prepare 3 parts of PVP and 30 parts of ethanol, heat to 40°C and stir for 5 minutes, add 1 part of p-toluenesulfonate adenosylmethionine, and mix;

[0049] (6) Granulation: The above suspension and soluble granules are mixed in a ratio of 4:6, and stirred in a rapid stirring pot for 10 min. The wet granules are quickly placed in a centrifugal coating granulator, and the main engine speed is adjusted to 120 r / min. The granules are spheronized for 30 min. During this period, 10% of the wetting agent is sprayed every 10 min. After that, the mixture is dried at a hot air temperature of 50°C and a blowing flow rate of 20 L / min for 5 h. The compound Dendrobium officinale granule preparation for clearing the liver and improving eyesight is obtained.

[0050] Comparative Example 1

[0051] The difference between this comparative example and Example 1 is that the Boc-L-cyclohexylalanine in step (3) is Boc-L-alanine, and the rest is the same as Example 1.

[0052] Comparative Example 2

[0053] The difference between this comparative example and Example 1 is that the amount of Boc-L-cyclohexylalanine added in this comparative example is different, and the specific amino acid modification steps are: 7 parts of Boc-L-cyclohexylalanine are dissolved in 40 parts of THF solution, cooled in an ice bath and stirred for 30 minutes, 3 parts of the above-mentioned filter residue insoluble substance are added, and the reaction is continued to be stirred for 2 hours at room temperature, filtered, and the filtrate is concentrated and purified by silica gel column chromatography, wherein the volume ratio of ethyl acetate: petroleum ether is 1:6, which is used as an eluent, and purified to obtain solid particles; the remaining implementation methods are the same as Example 1.

[0054] Comparative Example 3

[0055] The difference between this comparative example and Example 1 is that the amount of Boc-L-cyclohexylalanine added in this comparative example is different, and the specific amino acid modification steps are: dissolving 1 part of Boc-L-cyclohexylalanine in 40 parts of THF solution, cooling in an ice bath and stirring for 30 minutes, adding 3 parts of the above-mentioned filter residue insoluble substance, continuing to stir and react at room temperature for 2 hours, filtering, concentrating the filtrate and separating and purifying it by silica gel column chromatography, wherein the volume ratio of ethyl acetate: petroleum ether is 1:6, which is used as an eluent, purifying to obtain solid particles; the remaining implementation methods are the same as Example 1.

[0056] Comparative Example 4

[0057] The difference between this comparative example and Example 1 is that the monoethanolamide of dodecenoic acid in step (4) is alkanolamide, and the rest is the same as Example 1.

[0058] Comparative Example 5

[0059] The difference between this comparative example and Example 1 is that the amount of monoethanolamide of dodecenoic acid added in step (4) is different. The specific steps of the nanosuspension are as follows: 12 parts of poloxamer 188, 10 parts of sodium dodecyl sulfate and 14 parts of monoethanolamide of dodecenoic acid are added to 200 parts of distilled water, and stirred and decomposed for 10 minutes to obtain distilled water containing a surface suspending agent; 15 parts of the solid particles obtained in the previous step are weighed, added to the distilled water containing the surface suspending agent, and stirred and mixed thoroughly. After ultrasonication at 8000 r / min for 10 minutes, a coarse suspension is obtained, and the coarse suspension is circulated through a high-pressure homogenizer at a pressure of 1000 bar for 15 cycles for standby use; the rest is the same as in Example 1.

[0060] Comparative Example 6

[0061] The difference between this comparative example and Example 1 is that the amount of monoethanolamide of dodecenoic acid added in step (4) is different. The specific steps of the nanosuspension are as follows: 12 parts of poloxamer 188, 10 parts of sodium dodecyl sulfate, and 2 parts of monoethanolamide of dodecenoic acid are added to 200 parts of distilled water, and stirred and decomposed for 10 minutes to obtain distilled water containing a surface suspending agent; 15 parts of the solid particles obtained in the previous step are weighed, added to the distilled water containing the surface suspending agent, and stirred and mixed thoroughly. After ultrasonication at 8000 r / min for 10 minutes, a coarse suspension is obtained, and the coarse suspension is circulated through a high-pressure homogenizer at a pressure of 1000 bar for 15 cycles for standby use; the rest is the same as in Example 1.

[0062] Comparative Example 7

[0063] The difference between this comparative example and Example 1 is that the adenosylmethionine p-toluenesulfonate in step (5) is methionine, and the rest is the same as Example 1.

[0064] Comparative Example 8

[0065] The difference between this comparative example and Example 1 is that the amount of adenosylmethionine p-toluenesulfonate added in step (5) is different. The specific steps for preparing the wetting agent are: prepare 4 parts of PVP and 25 parts of ethanol, heat to 40° C. and stir for 5 minutes, add 1.5 parts of adenosylmethionine p-toluenesulfonate, and mix; the rest is the same as Example 1.

[0066] Comparative Example 9

[0067] The difference between this comparative example and Example 1 is that the amount of adenosylmethionine p-toluenesulfonate added in step (5) is different. The specific steps for preparing the wetting agent are: prepare 4 parts of PVP and 25 parts of ethanol, heat to 40° C. and stir for 5 minutes, add 0.5 parts of adenosylmethionine p-toluenesulfonate, and mix; the rest is the same as Example 1.

[0068] 1. Experimental Animals

[0069] 104 healthy male C57BL / 6 mice with a body weight of 19-23g were selected and purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. After one week of adaptive feeding, they were divided into a control group and a model group, with 8 mice in each group. 5g of the sample particles prepared in the corresponding embodiment and the comparative example were dissolved in 50ml of distilled water and given to the model group mice by gavage, and the control group was given an equal amount of sodium chloride saline, once a day, and fasted but not watered for 12h after feeding for 1 week, and the mice were killed after blood was taken, and about 1mL of blood was taken from the mouse eyeball, placed in a 1.5mL centrifuge tube, centrifuged at 3000r / min for 10min, and the supernatant was taken. The levels of AST and ALT in serum were determined by an ELISA instrument, and the operation was strictly in accordance with the instructions for the assay kit. The results are shown in Table 1 below.

[0070] Table 1 Changes of AST and ALT in mouse serum

[0071] Group AST(U / L) ALT(U / L) Control group 232.22±46.90 165.24±10.52 Model Example 1 94.56±52.98 54.31±8.51 Model Example 2 96.84±46.92 63.26±10.25 Model Example 3 Group 99.26±10.75 59.23±27.37 Model comparison group 1 147.58±9.87 165.37±13.46 Model comparison group 2 123.42±14.68 122.53±10.86 Model comparison group 3 143.21±16.87 156.46±14.87 Model comparison group 4 126.75±21.94 88.48±11.53 Model comparison group 5 118.79±31.85 96.58±15.94 Model comparison group 6 156.84±16.84 168.93±16.84 Model comparison group 7 116.98±15.76 112.54±20.53 Model comparison group 8 143.62±16.82 124.79±14.86 Model comparison group 9 164.73±16.97 168.75±±12.69

[0072] 2. Dissolution Test

[0073] Test sample: the liver-clearing and eyesight-improving Dendrobium officinale granules in Examples 1-3 and Comparative Examples 4-6.

[0074] Determination method: Refer to the second method of slurry method in Article 0931 of the "General Rules of the Four Parts of the 2020 Edition of the Chinese Pharmacopoeia". Specifically include the following steps: put 4g of the test sample into 100mL of water, 100r / min, take 20mL of samples at 1min, 2min, 5min, and 8min respectively, filter through a 0.45μm membrane, discard 10mL of the initial filtrate, and use the subsequent filtrate as the test solution. Use UV-visible spectrophotometry to measure the absorbance of each solution at 362nm, calculate the solubility, and obtain the following Table 2:

[0075] Table 2 Dissolution

[0076]

[0077]

[0078] 3. Powder wettability test:

[0079] The wettability determination method is as follows: take 100±1ml of 342ppm hard water (0.304g of anhydrous calcium chloride, 0.139g of six crystallized water magnesium chloride, diluted to 1 liter with distilled water), inject it into a 250ml beaker, and place the beaker in a constant temperature water bath at 25±1°C so that the liquid level is flush with the horizontal plane of the water bath. When the hard water reaches 25±1°C, weigh 5±0.1g of the sample with a watch glass, and pour all the samples evenly on the liquid surface of the beaker from a position flush with the mouth of the beaker, but do not disturb the liquid surface excessively. When adding the sample, use a stopwatch to time it immediately until the sample is completely wetted. Note the wetting time. Repeat this three times, take the average value, and use it as the wetting time of the sample. The wettability tests were carried out on Examples 1-3 and Comparative Examples 1-9, respectively. The results are shown in Table 3 below:

[0080] Table 3 Wettability results

[0081] Group Wettability / s Example 1 61 Example 2 64 Example 3 68 Comparative Example 1 79 Comparative Example 2 72 Comparative Example 3 71 Comparative Example 4 85 Comparative Example 5 75 Comparative Example 6 72 Comparative Example 7 79 Comparative Example 8 73 Comparative Example 9 75

Claims

1. A preparation process of a liver-clearing and eye-improving Dendrobium officinale composite granule, characterized in that: The specific steps include: (1) Extraction and separation: Wash, crush and dry 60 parts of Dendrobium officinale, 80 parts of Ginseng, 20 parts of Chinese Yam, 60 parts of Poria, 20 parts of Licorice, 20 parts of Cistanche, 40 parts of Lycium barbarum, 20 parts of Cuscuta, 60 parts of Rehmannia root, 30 parts of Schisandra chinensis, 80 parts of Asparagus cochinchinensis, 50 parts of Ophiopogon japonicus, 25 parts of Apricot kernel, 30 parts of Saposhnikovia divaricata, 30 parts of Chuanxiong, 20 parts of Citrus aurantium, 20 parts of Coptis chinensis, 35 parts of Cyathula officinalis, 25 parts of Chrysanthemum officinale. 20 parts of Tribulus terrestris, 20 parts of Coptidis amaranth seeds, 35 parts of Cassia seeds, 50 parts of buffalo horn concentrated powder, and 220-260 parts of antelope horn, and crush them through a 50-60 mesh sieve; put the mixed powder into an extraction kettle, and perform supercritical CO2 extraction, with a critical temperature of 30-35°C, a critical pressure of 7-9Mpa, an extraction time of 4-6h, an entrainer of anhydrous ethanol, and an entrainer dosage of 20%, to obtain an extract and a filter residue with insoluble substances; (2) The extract is separated and purified using d101b weakly polar macroporous resin, and then dialyzed in running water for 24 hours at a dialysis temperature of 30°C, with the water changed every 4 hours; then vacuum concentrated at 45-55°C to 1 / 10-1 / 3 of the original volume, 1-3 times of ethanol is added to the concentrate for precipitation, separated using Sephadex G75 gel, and finally freeze-dried at -70-78°C in a vacuum for 12-24 hours to obtain soluble particles; (3) Amino acid modification: 2-4 parts of an amino acid derivative are dissolved in 40 parts of a THF solution, cooled in an ice bath and stirred for 30 min, 2-4 parts of the above-mentioned insoluble substance in the filter residue are added, and the reaction is continued with stirring at room temperature for 2 h, filtered, and the filtrate is concentrated and purified by silica gel column chromatography, wherein the volume ratio of ethyl acetate to petroleum ether is 1:6, and it is used as an eluent for purification to obtain solid particles; (4) Nanosuspension: Weigh 10-20 parts of the above solids, add them into distilled water containing the surface suspending agent, stir and mix thoroughly, and ultrasonicate at 8000 r / min for 10 min to obtain a coarse suspension. Pass the coarse suspension through a high-pressure homogenizer at a pressure of 1000 bar for 15 cycles and set aside; (5) Granulation: The above suspension and soluble granules are mixed in a ratio of 4:6, stirred in a rapid stirring pot for 10-15 minutes, and then the wet granules are quickly placed in a centrifugal coating granulator, the main engine speed is adjusted to 100-120r / min, and spheronization is performed for 30 minutes. During this period, 10% of the wetting agent is sprayed every 10 minutes, and then dried for 5 hours at a hot air temperature of 50-60°C and a blast flow rate of 20L / min to obtain the compound Dendrobium officinale granule preparation for clearing the liver and improving eyesight; The amino acid derivative is Boc-L-cyclohexylalanine; The preparation of the distilled water containing the surface suspending agent in step (4) is as follows: 10-15 parts of poloxamer 188, 8-12 parts of sodium dodecyl hydrogen sulfate, and 5-8 parts of a stabilizing agent are added to 200 parts of distilled water, and stirred for decomposition for 10 minutes; The stabilizing regulator is monoethanolamide of dodecenoic acid.

2. The preparation process of the composite particles of Dendrobium officinale for clearing liver and improving eyesight as claimed in claim 1, characterized in that: The preparation of the wetting agent in step (5) is as follows: prepare 3-5 parts of PVP and 20-30 parts of ethanol, heat to 40° C. and stir for 5 minutes, add 0.5-1 part of a wetting regulator, and mix.

3. The preparation process of the composite particles of Dendrobium officinale for clearing liver and improving eyesight as claimed in claim 2, characterized in that: in, The wetting regulator is adenosylmethionine p-toluenesulfonate.

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