Tiamulin fumarate enteric-coated granules with high dissolution rate and efficient absorption capacity as well as preparation method and application of tiamulin fumarate enteric-coated granules

Abstract

The invention provides tiamulin fumarate enteric-coated granules with high dissolution rate and efficient absorption capacity as well as a preparation method and application of the tiamulin fumarate enteric-coated granules, and belongs to the technical field of pharmacy. The enteric-coated granules consist of tiamulin fumarate granules and a coating, wherein the tiamulin fumarate granules comprise tiamulin fumarate, microcrystalline cellulose, lactose, corn starch and termitomyces albuminosus polysaccharide B. The invention further discloses a preparation method of the tiamulin fumarate enteric-coated granules. The termitomyces albuminosus polysaccharide B is prepared through specific extraction and grading, the dissolution rate of the granules is remarkably improved, and efficient absorption of the medicine in the intestinal tract is promoted. Meanwhile, pharmacokinetic tests show that the enteric-coated granules have longer drug effect maintenance time and higher bioavailability. In addition, when the medicine is used for treating respiratory diseases of livestock and poultry, the cure rate of the medicine prepared by the invention is remarkably improved.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and in particular relates to enteric-coated tiamulin fumarate granules with high solubility and high absorption capacity. Background Art

[0002] Tiamulin fumarate is a bacteriostatic antibiotic belonging to the pleuromutilin class, which has the characteristic of producing bactericidal effects on sensitive bacteria at high concentrations. Similar to macrolide antibiotics, Tiamulin fumarate inhibits bacterial protein synthesis by binding to the 50S subunit of bacterial ribosomes. The drug shows good antibacterial activity against a variety of Gram-positive bacteria, including most staphylococci and streptococci (except group D streptococci), various mycoplasmas, and hyodysenteritis treponema. Tiamulin fumarate is mainly used to treat swine mycoplasma pneumonia, Haemophilus parasuis infection, infectious pleuropneumonia, and dysentery caused by swine treponema. It is also suitable for chronic respiratory diseases, mycoplasma pneumonia, infectious rhinitis, Escherichia coli disease, and duck infectious serositis in poultry.

[0003] At present, tiamulin fumarate is mainly administered orally. However, due to its bitter taste, it often causes adverse reactions such as anorexia, nausea and vomiting in livestock and poultry after use, which in turn affects the therapeutic effect. In addition, tiamulin fumarate is easily destroyed by gastric acid in the stomach of livestock and poultry, significantly reducing its therapeutic effect. Therefore, it is very necessary to develop a tiamulin fumarate enteric-coated granule that can be quickly released in the intestine and promote absorption, thereby prolonging the efficacy and improving its bioavailability in the body. Summary of the invention

[0004] The purpose of the present invention is to provide a kind of enteric-coated granules of tiamulin fumarate and a preparation method and application thereof, so that the granules can be better dissolved and absorbed in the intestine.

[0005] To achieve the above object, the present invention improves the following technical solutions:

[0006] Firstly, the present invention provides a fumarate tiamulin enteric-coated granule with high solubility and high absorption capacity, wherein the fumarate tiamulin enteric-coated granule is composed of fumarate tiamulin granules and a coating;

[0007] The raw materials for preparing the fumarate tiamulin granules consist of 15-25wt% fumarate tiamulin, 6wt% microcrystalline cellulose, 25-35wt% lactose, 35-38wt% corn starch, 1.5-5wt% black-skinned Alpinia fasciata polysaccharide, 2wt% sodium carboxymethyl starch and 2wt% hydroxypropyl methylcellulose solution.

[0008] Preferably, the raw materials for preparing the fumarate tiamulin granules consist of 20wt% fumarate tiamulin, 6wt% microcrystalline cellulose, 30wt% lactose, 38wt% corn starch, 2wt% Alpinia mellifera polysaccharide, 2wt% sodium carboxymethyl starch and 2wt% hydroxypropyl methylcellulose solution.

[0009] Preferably, the mass of the coating is 15% of the mass of the tiamulin fumarate particles;

[0010] The raw materials for preparing the coating consist of 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc powder, 0.9 wt % of polyethylene glycol and 82 wt % of water.

[0011] Preferably, the method for preparing the polysaccharide of Alpinia cerasifera comprises the following steps:

[0012] (a) cleaning the black skinned Alpinia hortensis, draining the water for later use;

[0013] (b) adding water at a mass ratio of 1:15 of Alpinia cerevisiae to water to obtain a mixture, and placing the mixture in a crusher for crushing and slurrying until a uniform slurry liquid is formed;

[0014] (c) adding the slurry into a heating container, heating to 120° C., continuously cooking for 3 hours, and cooling to room temperature;

[0015] (d) after filtering, centrifuging to obtain a total polysaccharide solution of Alpinia cerasifera;

[0016] (e) adding ethanol to the total polysaccharide solution of Alpinia thunbergii to make its final concentration be 80%, standing at 4° C. for 24 h, collecting the precipitate by centrifugation, placing it in a freeze dryer, and drying it to obtain the total polysaccharide of Alpinia thunbergii;

[0017] (f) adding water to the total polysaccharide of Alpinia thunbergii to prepare a total polysaccharide solution of Alpinia thunbergii;

[0018] (g) using a 12,000 Da ultrafiltration membrane to remove the retentate 1, collecting the filtrate 1, passing the filtrate 1 through an 8,000 Da ultrafiltration membrane, and collecting the filtrate 2;

[0019] (h) passing the filtrate 2 through a 5,000 Da ultrafiltration membrane, collecting the retentate 2, and obtaining a polysaccharide solution of Alpinia cerasifera;

[0020] (i) freeze-drying the polysaccharide solution of Alpinia cerana var.

[0021] Preferably, the preparation method of corydalis carboxylic acid tiamulin enteric-coated granules comprises the following steps:

[0022] (1) mixing tiamulin fumarate, microcrystalline cellulose, lactose, corn starch, black-skinned Alpinia fasciata polysaccharide, and sodium carboxymethyl starch uniformly;

[0023] (2) After passing through the drug sieve, hydroxypropyl methylcellulose solution is added to make a soft material, and then the raw granules are made by an extrusion spheronization machine;

[0024] (3) drying the granules in a forced air drying oven to obtain tiamulin fumarate granules;

[0025] (5) mixing acrylic resin copolymer, talc, polyethylene glycol and water to obtain a coating solution;

[0026] (6) The coating liquid is added to the coating pot of the extrusion spheronizer, and the tiamulin fumarate granules are added. The coating is performed to increase the coating weight by 15%. After the coating is completed, the granules are aged for 30 minutes to obtain the tiamulin fumarate enteric-coated granules.

[0027] Secondly, the present invention provides a method for preparing enteric-coated granules of tiamulin fumarate with high solubility and high absorption capacity, and the preparation method comprises the following method:

[0028] (1) 20 wt % of tiamulin fumarate, 6 wt % of microcrystalline cellulose, 30 wt % of lactose, 38 wt % of corn starch, 2 wt % of Alternaria japonica polysaccharide B, and 2 wt % of sodium carboxymethyl starch were mixed evenly;

[0029] (2) After passing through a 200-mesh sieve, 2 wt% hydroxypropyl methylcellulose solution was added to make a soft material, which was then made into plain granules using an extrusion spheronizer;

[0030] (3) drying the granules in a forced air drying oven at 60° C. for 2 hours to obtain tiamulin fumarate granules;

[0031] (4) mixing 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water to obtain a coating solution;

[0032] (5) The coating liquid is added to the coating pot of the extrusion spheronizer, and the tiamulin fumarate granules are added. The coating is performed to increase the coating weight by 15%. After the coating is completed, the granules are aged for 30 minutes to obtain the tiamulin fumarate enteric-coated granules.

[0033] Preferably, the Alpinia cerana polysaccharide is prepared by the above-mentioned method for preparing Alpinia cerana polysaccharide.

[0034] Secondly, the present invention provides the application of Alpinia thunbergii polysaccharide to improve the dissolution rate and absorption capacity of tylosin fumarate enteric granules, wherein the Alpinia thunbergii polysaccharide is prepared by the above-mentioned preparation method of Alpinia thunbergii polysaccharide.

[0035] Preferably, the tiamulin fumarate enteric-coated granules are prepared by the above-mentioned preparation method.

[0036] In addition, the present invention provides the use of black-skinned Alpinia thunbergii polysaccharide in the preparation of fumarate tyomycin enteric-coated granules for improving the therapeutic ability of chicken respiratory diseases, characterized in that the black-skinned Alpinia thunbergii polysaccharide is prepared by the preparation method of black-skinned Alpinia thunbergii polysaccharide described in claim 4;

[0037] The chicken respiratory disease is a chicken respiratory disease caused by mycoplasma.

[0038] The beneficial effects of the present invention are:

[0039] The fumarate tylosin enteric-coated granules of the present invention can effectively avoid the degradation of the drug in the gastric acid environment through coating technology, achieve sufficient dissolution in the intestine, and ensure efficient absorption of the drug. The addition of black-skinned Alpinia fasciata polysaccharide B further enhances the dissolution of the granules, making the dissolution of the drug in the intestinal fluid significantly higher than that of traditional preparations, thereby improving the bioavailability of the drug.

[0040] The addition of polysaccharide B of Alpinia fasciata to the enteric-coated tiamulin fumarate granules not only improves the dissolution efficiency of the drug, but also helps to increase the penetration of the solvent, thereby enhancing the efficacy. The granules prepared by the present invention can maintain a higher blood drug concentration and a longer maintenance time in the body. The pharmacokinetic test shows that the elimination half-life is longer and the area under the drug-time curve (AUC) increases, which proves that the drug effect is more lasting and the bioavailability in the body is higher.

[0041] In practical applications, the enteric-coated granules of the present invention show a higher cure rate and effective rate in treating poultry respiratory diseases (such as mycoplasma infection). Experimental data show that the drug treatment effect of the experimental group is significantly better than that of the control group. DETAILED DESCRIPTION

[0042] Now, various exemplary embodiments of the present invention are described in detail, and this detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention. It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention.

[0043] Example 1

[0044] 1. Mix 20wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 30wt% of lactose, 40wt% of corn starch and 2wt% of sodium carboxymethyl starch evenly;

[0045] 2. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronization machine to make plain granules;

[0046] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0047] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0048] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0049] Example 2

[0050] 1. Mix 20wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 30wt% of lactose, 38wt% of corn starch, 2wt% of lentinan (CAS: 37339-90-5), and 2wt% of sodium carboxymethyl starch uniformly;

[0051] 2. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronization machine to make plain granules;

[0052] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0053] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0054] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0055] Example 3

[0056] 1. Mix 20wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 30wt% of lactose, 38wt% of corn starch, 2% of astragalus polysaccharide (CAS: 84687-43-4), and 2wt% of sodium carboxymethyl starch uniformly;

[0057] 3. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronizer to make plain granules;

[0058] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0059] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0060] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0061] Example 4

[0062] 1. Mix 20wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 30wt% of lactose, 38wt% of corn starch, 2wt% of Alpinia fasciata polysaccharide A, and 2wt% of sodium carboxymethyl starch uniformly;

[0063] 3. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronizer to make plain granules;

[0064] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0065] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0066] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0067] Example 5

[0068] 1. Mix 20wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 30wt% of lactose, 38wt% of corn starch, 2wt% of Alpinia arborescens polysaccharide B, and 2wt% of sodium carboxymethyl starch uniformly;

[0069] 2. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronization machine to make plain granules;

[0070] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0071] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0072] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0073] Example 6

[0074] 1. Mix 20wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 30wt% of lactose, 38wt% of corn starch, 2wt% of Alpinia fasciata polysaccharide C, and 2wt% of sodium carboxymethyl starch uniformly;

[0075] 3. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronization machine to make plain granules;

[0076] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0077] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0078] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0079] Example 7

[0080] 1. Evenly mix 15wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 35wt% of lactose, 37.5wt% of corn starch, 2.5wt% of Alpinia arborescens polysaccharide B, and 2wt% of sodium carboxymethyl starch;

[0081] 2. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronization machine to make plain granules;

[0082] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0083] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0084] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0085] Example 8

[0086] 1. Mix 20wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 30wt% of lactose, 38.5wt% of corn starch, 1.5% of Alpinia fasciata polysaccharide B, and 2wt% of sodium carboxymethyl starch uniformly;

[0087] 2. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronization machine to make plain granules;

[0088] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0089] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0090] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0091] Example 9

[0092] 1. Mix 25wt% of tiamulin fumarate, 6wt% of microcrystalline cellulose, 25wt% of lactose, 35wt% of corn starch, 5wt% of Alpinia cerana polysaccharide B, and 2wt% of sodium carboxymethyl starch uniformly;

[0093] 2. After passing through a 200-mesh sieve, add 2 wt% hydroxypropyl methylcellulose solution to make a soft material, and use an extrusion spheronization machine to make plain granules;

[0094] 3. Dry the granules in a forced air drying oven at 60°C for 2 hours to obtain tiamulin fumarate granules;

[0095] 4. 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water were mixed to obtain a coating solution;

[0096] 5. Add the coating liquid into the coating pot of the extrusion spheronizer, add the fumarate tiamulin granules, and coat them with a coating weight gain of 15%. After coating, age for 30 minutes to obtain fumarate tiamulin enteric granules.

[0097] Example 10

[0098] 1. Choose fresh black-skinned chicken mushrooms, make sure they are free of mold and rot, wash them carefully with clean water, remove surface dirt and impurities, drain the water and set aside.

[0099] 2. Add water according to the mass ratio of black skin Alpinia fasciata to water of 1:15, and put the mixture into a crusher for crushing and slurrying until a uniform slurry liquid is formed;

[0100] 2. Add the slurry into a high pressure cooking pot, heat to 120°C, continue cooking at this temperature for 3 hours, and cool to room temperature;

[0101] 3. After filtering, centrifuge at 4000 rpm for 30 minutes to obtain a total polysaccharide solution of Alpinia cerasifera;

[0102] 4. Collect the supernatant into a new container and measure the volume of the supernatant, add ethanol to make the final concentration 80%, place it in a 4°C refrigerator for 24 hours, centrifuge it at 4000 rpm for 30 minutes, collect the precipitate, put it in a freeze dryer, and dry it to obtain the total polysaccharide of Aconitum fasciata;

[0103] 5. Dissolve the obtained total polysaccharide powder in an appropriate amount of purified water to prepare a polysaccharide solution with a concentration of 100 mg / mL;

[0104] 6. Using a 12,000Da ultrafiltration membrane, remove the retentate 1, collect the filtrate 1, pass the filtrate 1 through an 8,000Da ultrafiltration membrane, collect the retentate 2 and the filtrate 2, the retentate 2 is a polysaccharide with a molecular weight between 8,000Da and 12,000Da, called the black-skinned Alpinia fasciata polysaccharide A solution;

[0105] 8. Pass the filtrate 2 through a 5,000Da ultrafiltration membrane, collect the filtrate 3 and the retentate 3, the retentate 3 is a polysaccharide with a molecular weight between 5,000Da and 8,000Da, called the black-skinned Alpinia thunbergii polysaccharide B solution, and the filtrate 3 is a polysaccharide with a molecular weight less than 5,000Da, called the black-skinned Alpinia thunbergii polysaccharide C solution;

[0106] 6. The obtained polysaccharide A, B and C solutions were respectively loaded into the tray of a freeze dryer, and freeze-dried at -50°C and a vacuum degree of 10Pa to obtain black-skinned Alpinia argenteus polysaccharide A, black-skinned Alpinia argenteus B and black-skinned Alpinia argenteus polysaccharide C.

[0107] Experimental test 1

[0108] Detection of the difference in solubility of fumaric acid tiamulin granules prepared by different methods in acid

[0109] Detection method: According to the second method of dissolution and release determination in the appendix of the 2020 edition of the "Pharmacopoeia of the People's Republic of China" (for enteric-coated preparations) and the "Technical Guidelines for Dissolution Tests of Ordinary Oral Solid Preparations for Veterinary Use", pH 1.0 hydrochloric acid solution (9 ml of concentrated hydrochloric acid diluted with water to 1000 ml of solution) was used to simulate gastric solution, and high performance liquid chromatography was used for content determination. Dissolution conditions: slurry method, temperature 37 ° C ± 0.5 ° C, medium volume 900 ml, speed 75 rpm. The solubility differences of fumaric acid tylosin granules prepared by different methods in acid were determined, and the results are shown in Table 1.

[0110] Table 1 Differences in the solubility of fumaric acid tiamulin granules prepared by different methods in gastric juice

[0111]

[0112] From the results in Table 1, it can be seen that compared with Example 1, the solubility of the fumaric acid tylosin granules prepared in Examples 2-6 in gastric juice is improved, and the improvements in Example 2, Example 3, Example 4 and Example 6 are relatively limited, indicating that the addition of Lentinan, Astragalus polysaccharide, Alpinia oxyphylla polysaccharide A, and Alpinia oxyphylla polysaccharide C, although can increase the solubility of fumaric acid tylosin granules in gastric juice to a certain extent, the effect is relatively limited.

[0113] Further observations show that the solubility of the fumarate tylosin granules added with the black-skinned Alpinia fasciata polysaccharide B is significantly improved compared with Example 1, and a solubility of nearly 90% can be achieved in 15 minutes.

[0114] The possible reason for the above results is that the polysaccharide B of Alpinia fasciata has a suitable molecular weight and polysaccharide structure. The appropriate molecular weight and structure enable polysaccharide B to form a loose three-dimensional network inside the particles, which helps to improve the porosity of the particles, increase solvent penetration, and promote dissolution.

[0115] Experimental test 2

[0116] Detection of the difference in solubility of tiamulin fumarate granules prepared by different methods in buffer

[0117] According to the second method of dissolution and release determination in the appendix of the 2020 edition of the Pharmacopoeia of the People's Republic of China (for enteric-coated preparations) and the method of the Technical Guidelines for Dissolution Test of Ordinary Oral Solid Preparations for Veterinary Use, pH 1.0 hydrochloric acid solution (9 ml of concentrated hydrochloric acid was diluted with water to 1000 ml of solution) and pH 6.8 phosphate buffer solution (a mixed solution of 750 ml of 0.1 mol / L hydrochloric acid and 250 ml of 0.2 mol / L sodium phosphate solution, adjusted to pH 6.8) were used to simulate gastric solution and intestinal solution respectively. The dissolution conditions were: slurry method, temperature 37 ° C ± 0.5 ° C, medium volume 900 ml, and speed 75 rpm. The dissolution differences of tylosin fumarate enteric-coated granules prepared by different methods in intestinal fluid were determined, and the results are shown in Table 2.

[0118] Table 2 Differences in the dissolution of tiamulin fumarate enteric-coated granules prepared by different methods in intestinal fluid

[0119]

[0120] From the results in Table 2, it can be seen that the dissolution of the enteric-coated granules of tiamulin fumarate in all examples at pH 1.0 within 120 minutes was less than 3%, which indicates that the enteric coating effectively prevents the premature release of the drug in gastric juice.

[0121] The dissolution rates of all examples increased with time, which is consistent with the release characteristics of enteric-coated preparations under intestinal pH conditions. The dissolution rates of the enteric-coated tiamulin fumarate granules of Example 5 were significantly higher than those of other experiments at each time point, especially within the first 30 minutes, when the dissolution rate was more significantly improved.

[0122] A detailed comparison revealed that the initial dissolution rate was: the dissolution rate of the enteric-coated granules of fumaric acid tiamulin in Example 5 reached 31.30% at 5 min, rapidly increased to 63.38% at 10 min, and reached 76.65% at 15 min. The dissolution rates of the enteric-coated granules of fumaric acid tiamulin in other examples were between 17.39% and 25.20% at 5 min, between 33.25% and 38.66% at 10 min, and between 51.50% and 64.13% at 15 min. It can be seen that the dissolution rate of the enteric-coated granules of fumaric acid tiamulin in Example 5 is much higher than that of the other examples, indicating that the drug prepared in Example 5 can be released faster in the intestinal fluid.

[0123] Overall dissolution: The dissolution rate of the enteric-coated tiamulin fumarate granules of Example 5 reached 95.78% at 60 min, which was close to complete release. The dissolution rates of the enteric-coated tiamulin fumarate granules of other examples were between 82.17% and 87.78%, which were significantly lower than those of Example 5. This shows that the enteric-coated tiamulin fumarate granules of Example 5 not only had a fast initial dissolution rate, but also had a higher final dissolution rate.

[0124] In general, the experimental results in Table 2 clearly show that the black-skinned Alpinia fasciata polysaccharide B significantly improves the dissolution of tylosin fumarate enteric granules. Compared with other polysaccharides or preparations without added polysaccharides, preparations with added polysaccharide B have a faster dissolution rate in intestinal fluid and a higher solubility.

[0125] Experimental test 3

[0126] Detection of the pharmacokinetics of the enteric-coated granules of tiamulin fumarate prepared by the present invention

[0127] Forty-five healthy broilers of 20 days old from the same batch were used. All broilers were fasted for 12 hours before administration. Each broiler was weighed and blood samples were collected as a blank control group.

[0128] The first group, No. 1-15, is the test drug group of tiamulin fumarate enteric-coated granules prepared in Example 1.

[0129] The second group, No. 16-30, is the test drug group of tiamulin fumarate enteric-coated granules prepared in Example 5;

[0130] The third group, No. 31-45, is a commercially available tylosin fumarate premix;

[0131] All drugs were administered by oral gavage at a dose of 50 mg / kg body weight;

[0132] About 1.5 ml of blood was collected from the subwing vein of each broiler at 0.25, 0.5, 1, 2, 3, 4, 6, 8, 12, 24, 36, 48, 72, and 96 h after administration. The collected blood samples were placed in blood collection tubes containing the anticoagulant heparin sodium, centrifuged at 3000 rpm for 10 min, plasma was separated, and the supernatant was frozen at -20°C for measurement;

[0133] Sample pretreatment: thaw the frozen plasma sample naturally, take 0.5 mL of plasma sample, place it in a 1.5 mL centrifuge tube, add 0.5 mL of acetonitrile of 4% phosphoric acid solution, ultrasonicate for 2 min, vortex mix for 2 min, centrifuge at 6000 rpm for 10 min, take out the supernatant, add 1 mL of 4% phosphoric acid solution to the residue, ultrasonicate for 2 min, vortex mix for 2 min, centrifuge at 6000 rpm for 10 min, take out the supernatant, combine it with the first supernatant, mix well, pass through the HLB column, wash with 1 mL of water, and then elute with 1 mL of methanol, collect the methanol solution, and obtain;

[0134] Mass spectrometry conditions: heated atmospheric pressure electrospray ion source, selective positive ion monitoring mode, evaporator temperature 300°C, gas flow rate 9 L / min, sheath gas (high purity nitrogen) pressure 25 psi, sheath gas temperature 350°C, sheath gas flow rate 10 L / min, spray voltage 4.0 Kv.

[0135] The results are shown in Table 3:

[0136] Table 3 Pharmacokinetic test results of tiamulin fumarate enteric-coated granules

[0137]

[0138] As can be seen from the results in Table 3, the pharmacokinetic test results of the enteric-coated tiamulin fumarate granules prepared in Example 5 are significantly better than those in Example 1 and the premix. This result shows that the preparation of Example 5 has obvious advantages in bioavailability and drug absorption. It shows that the polysaccharide B of Alpinia thunbergii may improve the intestinal absorption of drugs, allowing more drug molecules to enter the blood circulation through the intestinal wall. Adding polysaccharide B of Alpinia thunbergii to the enteric-coated granules enhances the protective effect of the drug and improves the physical and chemical stability of the preparation.

[0139] Experimental test 4

[0140] Detection of the therapeutic effect of the fumarate tiamulin enteric-coated granules prepared by the present invention on respiratory diseases caused by Mycoplasma galli

[0141] 400 20-day-old broilers were selected and clinically diagnosed by professional veterinarians (symptoms: decreased wheezing and breathing sounds, depression, loss of appetite, symptoms of nose shaking, runny nose) with respiratory diseases caused by mycoplasma, and were divided into three groups on average: 100 test drug group (I) (Example 5), 100 test drug group (II) (Example 1), 100 control drug group (commercially available premix) and 100 positive control group, and 100 healthy chickens of the same breed and age were selected as negative control group. The drugs were administered for three consecutive days and observed for 7 days, and the mortality rate, effective rate and cure rate were calculated.

[0142] Table 4 Grouping and treatment of experimental chickens

[0143]

[0144] Efficacy evaluation indicators:

[0145] Mortality rate: During the experimental period, the mortality rate of chickens that died of respiratory diseases caused by Mycoplasma galli was calculated.

[0146] Effectiveness: During the experimental period, the drug is continuously administered, and if the symptoms of chickens suffering from respiratory diseases caused by Mycoplasma galli are alleviated or disappear, the effectiveness (including the number of cured animals) is calculated.

[0147] Cure rate: After continuous administration of the drug, the spirit, feeding and drinking of chickens with respiratory diseases caused by Mycoplasma galli return to normal, and the respiratory symptoms completely disappear, and the cure rate is calculated.

[0148] Table 5 The therapeutic effect of drugs on respiratory diseases caused by Mycoplasma avium

[0149]

[0150] From the results in Table 5, it can be seen that compared with the control group and the experimental group (II), the effective rate of the experimental group (I) was significantly improved, indicating that feeding the fumarate tylosin enteric-coated granules containing black-skinned Alpinia fasciata polysaccharide B for the treatment of respiratory diseases caused by chicken Mycoplasma has a better therapeutic effect.

Claims

1. A fumarate tiamulin enteric-coated granule with high solubility and high absorption capacity, characterized in that: The tiamulin fumarate enteric-coated granules are composed of tiamulin fumarate granules and a coating; The raw materials for preparing the fumarate tiamulin granules consist of 15-25wt% fumarate tiamulin, 6wt% microcrystalline cellulose, 25-35wt% lactose, 35-38wt% corn starch, 1.5-5wt% black-skinned Alpinia fasciata polysaccharide, 2wt% sodium carboxymethyl starch and 2wt% hydroxypropyl methylcellulose solution.

2. The enteric-coated tiamulin fumarate granules according to claim 1, characterized in that: The raw materials for preparing the fumarate tiamulin granules consist of 20wt% fumarate tiamulin, 6wt% microcrystalline cellulose, 30wt% lactose, 38wt% corn starch, 2wt% black-skinned Alpinia fasciata polysaccharide, 2wt% sodium carboxymethyl starch and 2wt% hydroxypropyl methylcellulose solution.

3. The enteric-coated tiamulin fumarate granules according to claim 2, characterized in that: The mass of the coating is 15% of the mass of the fumarate tiamulin particles; The raw materials for preparing the coating consist of 15.3 wt % of acrylic resin copolymer, 1.8 wt % of talc powder, 0.9 wt % of polyethylene glycol and 82 wt % of water.

4. The enteric-coated tiamulin fumarate granules according to claim 3, characterized in that: The preparation method of the black-skinned Alpinia thunbergii polysaccharide comprises the following steps: (a) cleaning the black skinned Alpinia hortensis, draining the water for later use; (b) adding water at a mass ratio of 1:15 of Alpinia cerevisiae to water to obtain a mixture, and placing the mixture in a crusher for crushing and slurrying until a uniform slurry liquid is formed; (c) adding the slurry into a heating container, heating to 120° C., continuously cooking for 3 hours, and cooling to room temperature; (d) after filtering, centrifuging to obtain a total polysaccharide solution of Alpinia cerasifera; (e) adding ethanol to the total polysaccharide solution of Alpinia thunbergii to make its final concentration be 80%, standing at 4° C. for 24 h, collecting the precipitate by centrifugation, placing it in a freeze dryer, and drying it to obtain the total polysaccharide of Alpinia thunbergii; (f) adding water to the total polysaccharide of Alpinia thunbergii to prepare a total polysaccharide solution of Alpinia thunbergii; (g) using a 12,000 Da ultrafiltration membrane to remove the retentate 1, collecting the filtrate 1, passing the filtrate 1 through an 8,000 Da ultrafiltration membrane, and collecting the filtrate 2; (h) passing the filtrate 2 through a 5,000 Da ultrafiltration membrane, collecting the retentate 2, and obtaining a polysaccharide solution of Alpinia cerasifera; (i) freeze-drying the polysaccharide solution of Alpinia cerana var.

5. The enteric-coated tiamulin fumarate granules according to claim 4, characterized in that: The preparation method of corydalis carboxylic acid tiamulin enteric-coated granules comprises the following steps: (1) mixing tiamulin fumarate, microcrystalline cellulose, lactose, corn starch, black-skinned Alpinia fasciata polysaccharide, and sodium carboxymethyl starch uniformly; (2) After passing through the drug sieve, hydroxypropyl methylcellulose solution is added to make a soft material, and then the raw granules are made by an extrusion spheronization machine; (3) drying the granules in a forced air drying oven to obtain tiamulin fumarate granules; (5) mixing acrylic resin copolymer, talc, polyethylene glycol and water to obtain a coating solution; (6) The coating liquid is added to the coating pot of the extrusion spheronizer, and the tiamulin fumarate granules are added. The coating is performed to increase the coating weight by 15%. After the coating is completed, the granules are aged for 30 minutes to obtain the tiamulin fumarate enteric-coated granules.

6. A method for preparing enteric-coated tiamulin fumarate granules with high solubility and high absorption capacity, characterized in that: The preparation method comprises the following method: (1) 20 wt % of tiamulin fumarate, 6 wt % of microcrystalline cellulose, 30 wt % of lactose, 38 wt % of corn starch, 2 wt % of Alternaria japonica polysaccharide B, and 2 wt % of sodium carboxymethyl starch were mixed evenly; (2) After passing through a 200-mesh sieve, 2 wt% hydroxypropyl methylcellulose solution was added to make a soft material, which was then made into plain granules using an extrusion spheronizer; (3) drying the granules in a forced air drying oven at 60° C. for 2 hours to obtain tiamulin fumarate granules; (4) mixing 15.3 wt % of an acrylic resin copolymer, 1.8 wt % of talc, 0.9 wt % of polyethylene glycol and 82 wt % of water to obtain a coating solution; (5) The coating liquid is added to the coating pot of the extrusion spheronizer, and the tiamulin fumarate granules are added. The coating is performed to increase the coating weight by 15%. After the coating is completed, the granules are aged for 30 minutes to obtain the tiamulin fumarate enteric-coated granules.

7. The preparation method according to claim 6, characterized in that: The black-skinned Alpinia terrestris polysaccharide is prepared by the preparation method of the black-skinned Alpinia terrestris polysaccharide described in claim 4.

8. Application of Alpinia cerevisiae polysaccharide to improve the dissolution and absorption capacity of tiamulin fumarate enteric granules, characterized in that: The black-skinned Alpinia terrestris polysaccharide is prepared by the preparation method of the black-skinned Alpinia terrestris polysaccharide described in claim 4.

9. The use according to claim 8, characterized in that: The tiamulin fumarate enteric-coated granules are prepared by the preparation method described in claim 6.

10. The use of polysaccharide from Alpinia thunbergii in the preparation of enteric-coated tiamulin fumarate granules for improving the therapeutic ability of chicken respiratory diseases, characterized in that: The black-skinned Alpinia terrestris polysaccharide is prepared by the preparation method of the black-skinned Alpinia terrestris polysaccharide described in claim 4; The chicken respiratory disease is a chicken respiratory disease caused by mycoplasma.

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