Florfenicol-doxycycline compound preparation and preparation method thereof

Abstract

The invention relates to the technical field of pharmaceutical preparations, in particular to a florfenicol-doxycycline compound preparation and a preparation method thereof. The florfenicol-doxycycline compound preparation is prepared from the following raw materials in parts by mass: 5 to 10 parts of florfenicol, 5 to 20 parts of doxycycline hydrochloride, 20 to 40 parts of inclusion material, 8 to 14 parts of pullulan, 1 to 3 parts of nano silicon dioxide, 1 to 5 parts of glycerol, 1 to 2 parts of emulsifier, 0.1 to 0.3 part of genipin, 20 to 60 parts of filler, 0.1 to 1 part of surfactant and 0.1 to 1 part of lubricant. The florfenicol and the doxycycline hydrochloride are jointly applied, the effect of the florfenicol and the doxycycline hydrochloride tablet is better than that of a single component, meanwhile, the solubility of the florfenicol is improved through a granulation method and auxiliary materials, the florfenicol can be administrated through mixed feeding or drinking water, and the florfenicol and doxycycline hydrochloride tablet is convenient to clinically use.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and particularly relates to a florfenicol-doxycycline compound preparation and a preparation method thereof. Background Art

[0002] Doxycycline hydrochloride has high water solubility and liposolubility, is well absorbed orally, has a broad antibacterial spectrum and strong antibacterial activity, and is widely used in the treatment of bacterial diseases of the respiratory tract and digestive tract of livestock and poultry. Florfenicol has low water solubility and high liposolubility, and also has characteristics such as good absorption, wide distribution, and long half-life, and is commonly used in the treatment of Pasteurella and Escherichia coli infections. A number of studies have shown that the antibacterial effect is better when doxycycline hydrochloride and florfenicol are used in combination than when used alone. Experiments have proved that the cure rate of the combination of the two for pasteurellosis is as high as 93.33%, and the cure rate for chicken colibacillosis is the highest.

[0003] Chinese Patent CN201510785909.7 discloses a compound doxycycline hydrochloride florfenicol sustained-release microsphere suspension injection, which not only ensures the synergistic antibacterial effect, but also improves the stability of doxycycline hydrochloride and the solubility of florfenicol. Chinese Patent CN201110073072.5 discloses a veterinary compound florfenicol injection and a preparation method thereof, which expands the antibacterial spectrum, has a quick effect, reduces toxic and side effects, reduces the occurrence of drug resistance, does not affect the weight gain of pigs during and after treatment, and can treat both the symptoms and the root causes. Chinese Patent CN201210271104.7 discloses a compound florfenicol injection and a preparation method thereof, which has no cross-resistance with common antibacterial drugs, does not require grain for production, has a lower price than antibiotics with comparable efficacy, and has good efficacy for various types of infections, including mild, moderate, and severe infections. However, the injections disclosed in the above patents are inconvenient to use clinically and have high requirements for veterinarians.

[0004] Chinese Patent CN202310576806.4 discloses a doxycycline-florfenicol liposome, which solves the problems that doxycycline is easily oxidized when exposed to light and heat, and florfenicol is easily decomposed when heated. However, this method requires the use of organic solvents, which is likely to cause environmental pollution during production and has high production costs. Summary of the Invention

[0005] The purpose of the present invention is to solve the disadvantages existing in the prior art, and to propose a florfenicol-doxycycline compound preparation and a preparation method thereof.

[0006] A compound preparation of florfenicol and doxycycline, the raw materials of which include, by mass: 5 - 10 parts of florfenicol, 5 - 20 parts of doxycycline hydrochloride, 20 - 40 parts of clathrate material, 8 - 14 parts of pullulan polysaccharide, 1 - 3 parts of nano-silica, 1 - 5 parts of glycerol, 1 - 2 parts of emulsifier, 0.1 - 0.3 parts of genipin, 20 - 60 parts of filler, 0.1 - 1 part of surfactant, and 0.1 - 1 part of lubricant.

[0007] Preferably, the clathrate material is β-cyclodextrin or / and hydroxypropyl-β-cyclodextrin.

[0008] Preferably, the filler is at least one of anhydrous glucose, maltodextrin, and lactose.

[0009] Preferably, the surfactant is sodium dodecyl sulfate.

[0010] Preferably, the lubricant is talc or / and magnesium stearate.

[0011] Preferably, the emulsifier includes: span 80, tween 20.

[0012] Preferably, the mass ratio of span 80 to tween 20 is 5 - 6:1.

[0013] The preparation method of the above-mentioned compound preparation of florfenicol and doxycycline includes the following steps:

[0014] S1. Add the clathrate material and doxycycline hydrochloride into water and stir evenly, then add pullulan polysaccharide, nano-silica, and glycerol to it and perform ultrasonic treatment for 10 - 20 min to obtain the first system; mix the emulsifier and liquid paraffin evenly to obtain the second system;

[0015] S2. Continuously stir the second system, keep the system temperature at 40 - 45 °C, during the stirring process, dropwise add the first system to it, after the dropping is complete, continue to stir for 5 - 10 min, then dropwise add the phosphate buffer solution containing genipin, stir at 45 - 55 °C for 1.5 - 2.5 h, let it stand for 1 - 2 h, centrifuge to remove the oil phase, wash, and vacuum dry to obtain the doxycycline hydrochloride-loaded product;

[0016] S3. Granulate using florfenicol, the doxycycline hydrochloride-loaded product, filler, surfactant, and lubricant.

[0017] Preferably, in S1, the ultrasonic frequency is 20 - 25 kHz.

[0018] Preferably, in S2, the pH value of the phosphate buffer solution is 8.5.

[0019] Preferably, the granulation method in S3 is dry granulation, and the specific operation is as follows: Mix florfenicol, loaded doxycycline hydrochloride, filler, surfactant, and lubricant evenly; then granulate using a roller press with a feeding speed of 25 - 28 rpm, a roller speed of 8 - 10 rpm, a roller pressure of 2 - 3 MPa, a pre-granulation speed of 50 - 70 rpm, and a final granulation speed of 80 - 100 rpm, and screen the granules through a 20-mesh sieve.

[0020] Preferably, the granulation method in S3 is fluidized bed granulation, and the specific operation is as follows: Mix florfenicol, loaded doxycycline hydrochloride, surfactant, lubricant, and part of the filler evenly and place it in the fluidized bed as the substrate; dissolve the remaining filler in water to prepare the binder; add the binder to the fluidized bed for granulation with an inlet air temperature of 60 - 70 °C, a fan frequency of 10 - 15 Hz, a material temperature of 40 - 50 °C, a liquid spraying speed of 2 - 5 rpm, and an atomization pressure of 0.05 - 0.15 MPa.

[0021] Beneficial effects:

[0022] 1. In the present invention, the combined application of florfenicol and doxycycline hydrochloride has a better effect than the single use of either drug.

[0023] 2. The granulation method and excipients used in the present invention improve the solubility of florfenicol, and it can be administered by mixing in feed or drinking water, which is convenient for clinical use.

[0024] 3. The granulation method of the present invention has high granulation efficiency, low cost, and low pollution.

[0025] 4. The present invention realizes the efficient loading of doxycycline hydrochloride through the synergistic effect of cyclodextrin inclusion and polyacrylamide network embedding. The product is porous microparticles with the potential for slow release. It has been found through experiments that the compounding of loaded doxycycline hydrochloride and florfenicol not only has excellent preparation stability but also greatly enhances the stability at high temperature and high humidity.

[0026] 5. The present invention loads water-insoluble florfenicol and strongly polar doxycycline hydrochloride, which not only overcomes the disadvantage of the difficulty in blending water-soluble and fat-soluble drugs but also reduces the minimum inhibitory concentration against Escherichia coli, Staphylococcus aureus, and Pasteurella, prolongs the action time of the two drugs in animals, maintains a stable synergistic antibacterial effect in vivo, and has a significant clinical treatment effect. Description of the drawings

[0027] Figure 1 It is a comparison chart of the minimum inhibitory concentration of the preparation samples obtained in Example 5 and Comparative Examples 1 - 3 with 10% florfenicol powder and 10% doxycycline hydrochloride soluble powder.

[0028] Figure 2The figure shows the comparison of the body weights of broilers in the blank control group, model group, florfenicol group, doxycycline group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group.

[0029] Figure 3 The figure shows the comparison of the treatment effective rates and mortality rates of broilers in the blank control group, model group, florfenicol group, doxycycline group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group. Detailed implementation manners

[0030] The present invention will be further explained below in conjunction with specific embodiments.

[0031] Example 1

[0032] A florfenicol-doxycycline compound preparation, the raw materials thereof include: 5 g of florfenicol, 5 g of doxycycline hydrochloride, 20 g of β-cyclodextrin, 8 g of pullulan polysaccharide, 1 g of nano-silica, 1 g of glycerol, 1 g of emulsifier, 0.1 g of genipin, 20 g of anhydrous glucose, 0.1 g of sodium dodecyl sulfate, and 0.1 g of magnesium stearate. The emulsifier is composed of span 80 and tween 20 in a mass ratio of 5:1.

[0033] The preparation method of the above florfenicol-doxycycline compound preparation comprises the following steps:

[0034] S1. Add β-cyclodextrin and doxycycline hydrochloride to 100 g of deionized water and stir evenly, add pullulan polysaccharide, nano-silica, and glycerol thereto and perform ultrasonic treatment for 10 min, the ultrasonic frequency is 20 kHz, to obtain a first system; mix the emulsifier and 400 g of liquid paraffin evenly to obtain a second system;

[0035] S2. Continuously stir the second system, the stirring speed is 2000 r / min, keep the system temperature at 40 °C, dropwise add the first system thereto during the stirring process, continue to stir for 5 min after the dropwise addition is complete, then dropwise add a pH = 8.5 phosphate buffer solution containing genipin, stir at 45 °C for 1.5 h, stand for 1 h, centrifuge to remove the oil phase, and wash the product successively with absolute ethanol and deionized water, and perform vacuum drying to obtain the doxycycline hydrochloride-loaded product;

[0036] S3. Mix florfenicol, the doxycycline hydrochloride-loaded product, sodium dodecyl sulfate, magnesium stearate, and 8 g of anhydrous glucose evenly, and place them in a fluidized bed as a substrate; dissolve the remaining anhydrous glucose in 48 mL of purified water to prepare an adhesive; add the adhesive to the fluidized bed for granulation, the inlet air temperature is 60 °C, the fan frequency is 10 Hz, the material temperature is 40 °C, the liquid spraying speed is 2 rpm, and the atomization pressure is 0.05 MPa.

[0037] Example 2

[0038] A compound preparation of florfenicol and doxycycline, the raw materials of which include: 9 g of florfenicol, 20 g of doxycycline hydrochloride, 35 g of β-cyclodextrin, 14 g of pullulan polysaccharide, 3 g of nano-silica, 5 g of glycerol, 2 g of emulsifier, 0.3 g of genipin, 60 g of maltodextrin, 1 g of sodium dodecyl sulfate, and 1 g of talc powder. The emulsifier consists of span 80 and tween 20 in a mass ratio of 6:1.

[0039] The preparation method of the above-mentioned compound preparation of florfenicol and doxycycline includes the following steps:

[0040] S1. Add β-cyclodextrin and doxycycline hydrochloride to 200 g of deionized water and stir evenly. Add pullulan polysaccharide, nano-silica, and glycerol thereto and perform ultrasonic treatment for 20 min at an ultrasonic frequency of 25 kHz to obtain a first system; mix the emulsifier and 600 g of liquid paraffin evenly to obtain a second system.

[0041] S2. Continuously stir the second system at a stirring speed of 4000 r / min, keep the system temperature at 45 °C, dropwise add the first system during the stirring process, continue to stir for 10 min after the dropping is complete, then dropwise add a pH = 8.5 phosphate buffer solution containing genipin, stir at 55 °C for 2.5 h, let stand for 2 h, centrifuge to remove the oil phase, and wash the product successively with absolute ethanol and deionized water, and then perform vacuum drying to obtain the doxycycline hydrochloride-loaded product.

[0042] S3. Mix florfenicol, the doxycycline hydrochloride-loaded product, sodium dodecyl sulfate, talc powder, and 36 g of maltodextrin evenly and place them in a fluidized bed as a substrate; dissolve the remaining maltodextrin in 144 mL of purified water to prepare a binder; add the binder to the fluidized bed for granulation, with an inlet air temperature of 70 °C, a fan frequency of 15 Hz, a material temperature of 50 °C, a liquid spraying speed of 5 rpm, and an atomization pressure of 0.15 MPa.

[0043] Example 3

[0044] A compound preparation of florfenicol and doxycycline, the raw materials of which include: 7 g of florfenicol, 16 g of doxycycline hydrochloride, 25 g of hydroxypropyl-β-cyclodextrin, 12 g of pullulan polysaccharide, 1.5 g of nano-silica, 4 g of glycerol, 1.2 g of emulsifier, 0.25 g of genipin, 30 g of lactose, 0.7 g of sodium dodecyl sulfate, and 0.5 g of magnesium stearate. The emulsifier consists of span 80 and tween 20 in a mass ratio of 5.8:1.

[0045] The preparation method of the above-mentioned compound preparation of florfenicol and doxycycline includes the following steps:

[0046] S1. Add hydroxypropyl-β-cyclodextrin and doxycycline hydrochloride to 120 g of deionized water and stir evenly. Then add pullulan, nano-silica, and glycerol to it and perform ultrasonic treatment for 18 min at an ultrasonic frequency of 21 kHz to obtain the first system; Mix the emulsifier and 550 g of liquid paraffin evenly to obtain the second system;

[0047] S2. Continuously stir the second system at a stirring speed of 2500 r / min, keep the system temperature at 43 °C, dropwise add the first system during the stirring process. After the dropping is complete, continue stirring for 7 min, then dropwise add the pH = 8.5 phosphate buffer solution containing genipin, stir at 50 °C for 100 min, let it stand for 80 min, centrifuge to remove the oil phase, and wash the product successively with absolute ethanol and deionized water, and then dry it under vacuum to obtain the doxycycline hydrochloride-loaded product;

[0048] S3. Mix florfenicol, the doxycycline hydrochloride-loaded product, lactose, sodium dodecyl sulfate, and magnesium stearate evenly; Then granulate using a roller, with a feeding speed of 25 rpm, a roller speed of 8 rpm, a roller pressure of 2 MPa, a pre-granulation speed of 50 rpm, a final granulation speed of 80 rpm, and granulate through a 20-mesh sieve.

[0049] Example 4

[0050] A florfenicol-doxycycline compound preparation, the raw materials of which include: 9 g of florfenicol, 8 g of doxycycline hydrochloride, 35 g of hydroxypropyl-β-cyclodextrin, 10 g of pullulan, 2.5 g of nano-silica, 2 g of glycerol, 1.8 g of emulsifier, 0.15 g of genipin, 50 g of lactose, 0.5 g of sodium dodecyl sulfate, and 0.8 g of talc powder. The emulsifier is composed of span 80 and tween 20 in a mass ratio of 5.2:1.

[0051] The preparation method of the above-mentioned florfenicol-doxycycline compound preparation includes the following steps:

[0052] S1. Add hydroxypropyl-β-cyclodextrin and doxycycline hydrochloride to 180 g of deionized water and stir evenly. Then add pullulan, nano-silica, and glycerol to it and perform ultrasonic treatment for 12 min at an ultrasonic frequency of 24 kHz to obtain the first system; Mix the emulsifier and 450 g of liquid paraffin evenly to obtain the second system;

[0053] S2. Continuously stir the second system at a stirring speed of 3500 r / min, keep the system temperature at 41 °C, dropwise add the first system during the stirring process. After the dropping is complete, continue stirring for 9 min, then dropwise add the pH = 8.5 phosphate buffer solution containing genipin, stir at 50 °C for 140 min, let it stand for 100 min, centrifuge to remove the oil phase, and wash the product successively with absolute ethanol and deionized water, and then dry it under vacuum to obtain the doxycycline hydrochloride-loaded product;

[0054] S3. Mix florfenicol, loaded doxycycline hydrochloride, lactose, sodium dodecyl sulfate, and talc powder evenly; then granulate using a roller press with a feeding speed of 28 rpm, a roller speed of 10 rpm, a roller pressure of 3 MPa, a pre-granulation speed of 70 rpm, a final granulation speed of 100 rpm, and screen the granules through a 20-mesh sieve.

[0055] Example 5

[0056] A florfenicol-doxycycline compound preparation, the raw materials of which include: 10 g of florfenicol, 10 g of doxycycline hydrochloride, 40 g of hydroxypropyl-β-cyclodextrin, 11 g of pullulan polysaccharide, 2 g of nano-silica, 3 g of glycerol, 1.5 g of emulsifier, 0.2 g of genipin, 40 g of lactose, 0.3 g of sodium dodecyl sulfate, and 0.3 g of magnesium stearate. The emulsifier is composed of span 80 and tween 20 in a mass ratio of 5.5:1.

[0057] The preparation method of the above florfenicol-doxycycline compound preparation includes the following steps:

[0058] S1. Add hydroxypropyl-β-cyclodextrin and doxycycline hydrochloride to 150 g of deionized water and stir evenly. Add pullulan polysaccharide, nano-silica, and glycerol thereto and perform ultrasonic treatment for 15 min at an ultrasonic frequency of 22.5 kHz to obtain a first system; mix the emulsifier and 500 g of liquid paraffin evenly to obtain a second system;

[0059] S2. Continuously stir the second system at a stirring speed of 3000 r / min, keep the system temperature at 42 °C, dropwise add the first system thereto during the stirring process, continue stirring for 8 min after the dropping is complete, then dropwise add a pH = 8.5 phosphate buffer solution containing genipin, stir at 50 °C for 120 min, let stand for 90 min, centrifuge to remove the oil phase, and wash the product successively with absolute ethanol and deionized water, and then dry it under vacuum to obtain loaded doxycycline hydrochloride;

[0060] S3. Mix florfenicol, loaded doxycycline hydrochloride, lactose, sodium dodecyl sulfate, and magnesium stearate evenly; then granulate using a roller press with a feeding speed of 25 rpm, a roller speed of 8 rpm, a roller pressure of 2.5 MPa, a pre-granulation speed of 50 rpm, a final granulation speed of 80 rpm, and screen the granules through a 20-mesh sieve.

[0061] Comparative Example 1

[0062] A florfenicol-doxycycline compound preparation, the raw materials of which include: 10 g of florfenicol, 10 g of doxycycline hydrochloride, 40 g of hydroxypropyl-β-cyclodextrin, 57.8 g of lactose, 0.3 g of sodium dodecyl sulfate, and 0.3 g of magnesium stearate.

[0063] The preparation method of the above-mentioned florfenicol-doxycycline compound preparation comprises the following steps:

[0064] Mix florfenicol, doxycycline hydrochloride, hydroxypropyl-β-cyclodextrin, lactose, sodium lauryl sulfate, and magnesium stearate evenly; then granulate using a roller press with a feeding speed of 25 rpm, a roller speed of 8 rpm, a roller pressure of 2.5 MPa, a pre-granulation speed of 50 rpm, a final granulation speed of 80 rpm, and screen the granules through a 20-mesh sieve.

[0065] Comparative Example 2

[0066] A florfenicol-doxycycline compound preparation, the raw materials of which include: 10 g of florfenicol, 10 g of doxycycline hydrochloride, 51 g of hydroxypropyl-β-cyclodextrin, 2 g of nano-silica, 3 g of glycerol, 1.5 g of emulsifier, 40 g of lactose, 0.3 g of sodium lauryl sulfate, and 0.3 g of magnesium stearate. The emulsifier is composed of Span 80 and Tween 20 in a mass ratio of 5.5:1.

[0067] The preparation method of the above-mentioned florfenicol-doxycycline compound preparation comprises the following steps:

[0068] S1. Add hydroxypropyl-β-cyclodextrin and doxycycline hydrochloride to 150 g of deionized water and stir evenly. Add nano-silica and glycerol thereto and perform ultrasonic treatment for 15 min at an ultrasonic frequency of 22.5 kHz to obtain a first system; mix the emulsifier and 500 g of liquid paraffin evenly to obtain a second system.

[0069] S2. Continuously stir the second system at a stirring speed of 3000 r / min, keep the system temperature at 42 °C, dropwise add the first system thereto during the stirring process, continue stirring for 8 min after the dropping is complete, let it stand for 90 min, centrifuge to remove the oil phase, and wash the product successively with absolute ethanol and deionized water, and then dry it under vacuum to obtain the doxycycline hydrochloride-loaded product.

[0070] S3. Mix florfenicol, the doxycycline hydrochloride-loaded product, lactose, sodium lauryl sulfate, and magnesium stearate evenly; then granulate using a roller press with a feeding speed of 25 rpm, a roller speed of 8 rpm, a roller pressure of 2.5 MPa, a pre-granulation speed of 50 rpm, a final granulation speed of 80 rpm, and screen the granules through a 20-mesh sieve.

[0071] Comparative Example 3

[0072] A compound preparation of florfenicol and doxycycline, the raw materials of which include: 10 g of florfenicol, 10 g of doxycycline hydrochloride, 40 g of hydroxypropyl-β-cyclodextrin, 11 g of pullulan polysaccharide, 2 g of nano-silica, 3 g of glycerol, 0.2 g of genipin, 40 g of lactose, 0.3 g of sodium dodecyl sulfate, and 0.3 g of magnesium stearate.

[0073] The preparation method of the above-mentioned compound preparation of florfenicol and doxycycline includes the following steps:

[0074] S1. Add hydroxypropyl-β-cyclodextrin and doxycycline hydrochloride to 150 g of deionized water, stir evenly, add pullulan polysaccharide, nano-silica, and glycerol thereto, and perform ultrasonic treatment for 15 min at an ultrasonic frequency of 22.5 kHz to obtain a first system;

[0075] S2. Dropwise add a pH = 8.5 phosphate buffer solution containing genipin to the first system, stir at 50 °C for 120 min, stand for 90 min, wash the product successively with absolute ethanol and deionized water, and perform vacuum drying to obtain doxycycline hydrochloride loaded;

[0076] S3. Mix florfenicol, doxycycline hydrochloride loaded, lactose, sodium dodecyl sulfate, and magnesium stearate evenly; then granulate with a roller, the feeding speed is 25 rpm, the roller speed is 8 rpm, the pressure of the pressing wheel is 2.5 MPa, the pre-granulation speed is 50 rpm, the final granulation speed is 80 rpm, and granulate with a 20-mesh sieve.

[0077] Take 1.0 g of each of the preparation samples obtained in Example 5 and Comparative Examples 1-3, place them in a beaker containing 100 mL of purified water at 25 °C, and it can be observed that all the preparation samples are dissolved and the solution is clear when stirred at 400 rpm for 10 min.

[0078] Place the preparation samples obtained in Example 5 and Comparative Examples 1-3 in a constant temperature and humidity chamber at a temperature of 40 ± 2 °C and a relative humidity of 75 ± 5% for 6 months, take samples at 0, 1, 3, and 6 months respectively, and inspect the properties, loss on drying, and content of the samples. The results are shown in Table 1.

[0079] Table 1 Stability test of the compound preparation of florfenicol and doxycycline

[0080]

[0081]

[0082] It can be seen from the above results that the stability of the preparation sample obtained in Example 5 is better than that of Comparative Examples 1-3 (P < 0.05) in a constant temperature and humidity environment at a temperature of 40 ± 2 °C and a relative humidity of 75 ± 5%.

[0083] The preparation samples obtained in Example 5 and Comparative Examples 1-3 were placed in a constant temperature and humidity chamber at a temperature of 60 ± 2 °C for 10 days. Sampling was carried out on the 0th day, 5th day, and 10th day respectively, and the properties and contents of the samples were inspected. The results are shown in Table 2.

[0084] Table 2 High-temperature stability test of florfenicol-doxycycline compound preparation

[0085]

[0086] It can be seen from the above results that the stability of the preparation sample obtained in Example 5 is better than that of Comparative Examples 1-3 in a high-temperature environment of 60 ± 2 °C (P < 0.05).

[0087] The preparation samples obtained in Example 5 and Comparative Examples 1-3 were placed in a constant temperature and humidity chamber at normal temperature and a relative humidity of 90 ± 5% for 10 days. Sampling was carried out on the 0th day, 5th day, and 10th day respectively, and the properties and contents of the samples were inspected. The results are shown in Table 3.

[0088] Table 3 High-humidity stability test of florfenicol-doxycycline compound preparation

[0089]

[0090] It can be seen from the above results that the stability of the preparation sample obtained in Example 5 is the best in a high-humidity environment with a relative humidity of 90 ± 5%, but there is no significant difference compared with Comparative Examples 1-3 (P > 0.05).

[0091] The minimum inhibitory concentrations (MIC) (μg / mL) of the preparation samples obtained in Example 5 and Comparative Examples 1-3, 10% florfenicol powder, and 10% doxycycline hydrochloride soluble powder against Escherichia coli ATCC25922, Staphylococcus aureus ATCC25923, and Pasteurella multocida ATCC11859 were determined by the microbroth dilution method respectively.

[0092] The experimental results are as Figure 1 shown. The minimum inhibitory concentrations of the preparations obtained in Example 5 and Comparative Examples 1-3 are all lower than those of 10% florfenicol powder and 10% doxycycline hydrochloride soluble powder; and the minimum inhibitory concentrations of Comparative Example 1 are all the lowest, which should be due to the fact that Example 5 uses more excipients compared with Comparative Examples 2-3; however, the minimum inhibitory concentration of the preparation obtained in Example 5 is lower than that of Comparative Examples 2-3, and the antibacterial effect is better than that of Comparative Examples 2-3 (P < 0.05).

[0093] In a chicken farm in Zhejiang, 120 4-week-old AA broilers with similar body weights were taken, allowed to freely eat and drink for 1 week, and randomly divided into 8 groups, with 15 in each group, namely the blank control group, the model group, the florfenicol group, the doxycycline group, the group of Example 5, the group of Comparative Example 1, the group of Comparative Example 2, and the group of Comparative Example 3. Except for the blank control group, the remaining broilers were intraperitoneally injected with 0.3 mL of Escherichia coli at 1×10 9 CFU / mL to establish a chicken Escherichia coli disease model.

[0094] All groups were raised by the conventional feeding method. Among them: the florfenicol group was administered 10% florfenicol powder (mixed in drinking water, adding 1 g of 10% florfenicol powder to every 1 L of water), the doxycycline group was administered 10% doxycycline hydrochloride soluble powder (mixed in drinking water, adding 6 g of 10% doxycycline hydrochloride soluble powder to every 1 L of water), the group of Example 5 was administered the preparation obtained in Example 5 (mixed in drinking water, adding 3.5 g of the preparation obtained in Example 5 to every 1 L of water), the group of Comparative Example 1 was administered the preparation obtained in Comparative Example 1 (mixed in drinking water, adding 3.5 g of the preparation obtained in Comparative Example 1 to every 1 L of water), the group of Comparative Example 2 was administered the preparation obtained in Comparative Example 2 (mixed in drinking water, adding 3.5 g of the preparation obtained in Comparative Example 2 to every 1 L of water), and the group of Comparative Example 3 was administered the preparation obtained in Comparative Example 3 (mixed in drinking water, adding 3.5 g of the preparation obtained in Comparative Example 3 to every 1 L of water). The drug was administered continuously for 7 days. The broilers in each group were weighed, and the treatment effective rate and mortality were calculated.

[0095] As Figure 2 shown, the broilers in the group of Example 5 had the highest body weight, which was better than the florfenicol group, the doxycycline group, and the groups of Comparative Examples 1-3 (P < 0.05), and was similar to the body weight of the broilers in the blank control group, with no significant difference between the two (P > 0.05).

[0096] As Figure 3 shown, the treatment effective rates of the broilers in the group of Example 5 and the group of Comparative Example 3 were both 100%, which were better than the other groups (P < 0.05); while the mortality rates of the broilers in the group of Example 5 and the groups of Comparative Examples 1-3 were all 0, which were better than the model group, the florfenicol group, and the doxycycline group (P < 0.05).

[0097] The applicant believes that: this is because the present invention realizes the high-efficiency loading of doxycycline hydrochloride through the synergistic effect of cyclodextrin inclusion and polyacrylamide network embedding, and the product is porous microparticles with slow-release potential. By using the compound of loaded doxycycline hydrochloride and florfenicol, not only the stability of the preparation is excellent, but also the high-temperature and high-humidity stability is greatly enhanced. The present invention loads water-insoluble florfenicol and strongly polar doxycycline hydrochloride, which not only overcomes the disadvantage of the difficulty in blending water-soluble and fat-soluble drugs, but also reduces the minimum inhibitory concentration against Escherichia coli, Staphylococcus aureus, and Pasteurella, prolongs the action time of the two in animals, and maintains a stable synergistic antibacterial effect in vivo, with a significant clinical treatment effect.

[0098] As described above, it is only the preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, according to the technical solution and inventive concept of the present invention, making equivalent substitutions or changes should be covered within the protection scope of the present invention.

Claims

1. A florfenicol-doxycycline compound preparation, characterized in that: The raw materials include, by mass, 5-10 parts of florfenicol, 5-20 parts of doxycycline hydrochloride, 20-40 parts of inclusion material, 8-14 parts of pullulan, 1-3 parts of nano silicon dioxide, 1-5 parts of glycerol, 1-2 parts of emulsifier, 0.1-0.3 parts of genipin, 20-60 parts of filler, 0.1-1 parts of surfactant and 0.1-1 parts of lubricant.

2. The florfenicol-doxycycline compound preparation according to claim 1, characterized in that: The inclusion material is β-cyclodextrin or / and hydroxypropyl-β-cyclodextrin.

3. The florfenicol-doxycycline compound preparation according to claim 1, characterized in that: The filler is at least one of anhydrous glucose, maltodextrin and lactose.

4. The florfenicol-doxycycline compound preparation according to claim 1, characterized in that: The surfactant is sodium lauryl sulfate.

5. The florfenicol-doxycycline compound preparation according to claim 1, characterized in that: Lubricants are talc and / or magnesium stearate.

6. The florfenicol-doxycycline compound preparation according to claim 1, characterized in that: Emulsifiers include: Span 80, Twain 20.

7. The florfenicol-doxycycline compound preparation according to claim 6, characterized in that: The mass ratio of Span 80 to Tween 20 is 5-6:

1.

8. A method for preparing the florfenicol-doxycycline compound preparation according to any one of claims 1 to 7, characterized in that: The steps include: S1, adding the inclusion material and doxycycline hydrochloride to water and stirring evenly, adding pullulan, nano-silicon dioxide and glycerol thereto and ultrasonically treating for 10-20 minutes to obtain a first system; mixing an emulsifier and liquid paraffin evenly to obtain a second system; S2, the second system is continuously stirred, the system temperature is maintained at 40-45°C, the first system is added dropwise thereto during the stirring process, stirring is continued for 5-10 min after the addition is complete, and then a phosphate buffer containing genipin is added dropwise thereto, stirred at 45-55°C for 1.5-2.5 h, allowed to stand for 1-2 h, centrifuged to remove the oil phase, washed, and vacuum dried to obtain the loaded doxycycline hydrochloride; S3, granulation is performed using florfenicol, loaded doxycycline hydrochloride, fillers, surfactants and lubricants.

9. The method for preparing the florfenicol-doxycycline compound preparation according to claim 8, characterized in that: In S1, the ultrasound frequency is 20-25kHz.

10. The method for preparing the florfenicol-doxycycline compound preparation according to claim 8, characterized in that: In S2, the pH value of the phosphate buffer is 8.5.

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