A florfenicol-doxycycline compound preparation and a preparation method thereof

Abstract

The present application relates to the technical field of pharmaceutical preparation, and in particular to a florfenicol-doxycycline compound preparation and a preparation method thereof.The raw materials of the florfenicol-doxycycline compound preparation include, by mass fraction, 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.The present application combines the use of florfenicol and doxycycline hydrochloride, and the effect is better than that of single use;meanwhile, the granulation method and the auxiliary materials used improve the solubility of florfenicol, and the florfenicol-doxycycline compound preparation can be administered by mixed feeding or drinking water, and is convenient for clinical use.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical preparations, in particular to a florfenicol-doxycycline compound preparation and a preparation method thereof. BACKGROUND

[0002] Doxycycline hydrochloride has high water solubility and fat solubility, good oral absorption, wide antibacterial spectrum and strong antibacterial activity, and is widely used for treating bacterial diseases of respiratory tract and digestive tract of livestock and poultry. Florfenicol also has low water solubility and high fat solubility, and also has good absorption, wide distribution, long half-life and other characteristics, and is often used for treating Pasteurella and Escherichia coli infections. A number of studies have shown that the antibacterial effect of doxycycline hydrochloride and florfenicol combined application is better than that of single use, and tests have shown that the cure rate of the combination for Pasteurella disease is as high as 93.33%, and the cure rate for chicken Escherichia coli disease 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 compound florfenicol injection for veterinary use and a preparation method thereof, which expands the antibacterial spectrum, has a quick effect, reduces toxic side effects and 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 cause. Chinese patent CN201210271104.7 discloses a compound florfenicol injection and a preparation method thereof, which has no cross-resistance with commonly used antibacterial drugs, does not require grain for production, has a lower price than antibiotics with comparable efficacy, and has good efficacy for light, medium and heavy infections. However, the injection disclosed in the above-mentioned patent is inconvenient to use in clinical practice and requires high requirements for veterinarians.

[0004] Chinese patent CN202310576806.4 discloses a doxycycline-florfenicol liposome, which solves the problems of easy oxidation of doxycycline under light and heat and easy decomposition of florfenicol under heat, but the method requires the use of organic solvents, which can easily cause environmental pollution during production and has high production cost. SUMMARY

[0005] The present application relates to the technical field of pharmaceutical preparations, in particular to a florfenicol-doxycycline compound preparation and a preparation method thereof.

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

[0007] Preferably, the inclusion material is beta-cyclodextrin or / and hydroxypropyl-beta-cyclodextrin.

[0008] Preferably, the filler is at least one of anhydrous glucose, malt dextrin, 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, Tween 20 is 5-6:1.

[0013] The preparation method of the above florfenicol-doxycycline compound preparation, comprising the following steps:

[0014] S1, the inclusion material, doxycycline hydrochloride is added to water and stirred uniformly, pullulan, nano-silicon dioxide, glycerol are added thereto and ultrasonically treated for 10-20 min to obtain a first system; the emulsifier and liquid paraffin are mixed uniformly to obtain a second system;

[0015] S2, continuously stirring the second system, keeping the system temperature at 40-45℃, adding the first system dropwise into the system during stirring, continuing to stir for 5-10 min after the dropwise addition is completed, then adding a phosphate buffer solution containing genipin dropwise into the system, stirring at 45-55℃ for 1.5-2.5 h, standing for 1-2 h, removing the oil phase by centrifugation, washing, vacuum drying to obtain doxycycline hydrochloride loaded;

[0016] S3, granulating florfenicol, doxycycline hydrochloride loaded, filler, surfactant, 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: the florfenicol, the loaded doxycycline hydrochloride, the filler, the surfactant and the lubricant are uniformly mixed; then the granulation is carried out by using a compression roller, the feeding rotation speed is 25-28 rpm, the compression roller rotation speed is 8-10 rpm, the pressure of the compression wheel is 2-3 MPa, the pre-granulation rotation speed is 50-70 rpm, the final granulation rotation speed is 80-100 rpm, and 20 mesh screen is used for granulation.

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

[0021] Beneficial effects:

[0022] 1. The florfenicol and the doxycycline hydrochloride are jointly used in the application, and the effect is better than that of single use.

[0023] 2. The granulation method and the excipient used in the application improve the solubility of the florfenicol, and the florfenicol can be administered by mixed feeding or drinking water, so that the clinical use is convenient.

[0024] 3. The granulation method has high granulation efficiency, low cost and small pollution.

[0025] 4. The florfenicol and the doxycycline hydrochloride are loaded by the synergistic effect of cyclodextrin inclusion and polyacrylamide network embedding, and the product is a porous microparticle with sustained release potential. It is found through experiments that: the florfenicol and the doxycycline hydrochloride are compounded, not only the preparation stability is excellent, but also the high temperature and high humidity stability is greatly enhanced.

[0026] 5. The water-insoluble florfenicol and the strong polar doxycycline hydrochloride are loaded, which not only overcomes the shortcomings that water-soluble and fat-soluble drugs are difficult to blend, but also reduces the minimum inhibitory concentration of escherichia coli, staphylococcus aureus and pasteurella multocida, prolongs the action time of the florfenicol and the doxycycline hydrochloride in the animal body, maintains stable synergistic antibacterial effect in the body, and has remarkable clinical treatment effect. BRIEF DESCRIPTION OF DRAWINGS

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

[0028] Figure 2The body weight comparison chart of the blank control group, the model group, the florfenicol group, the doxycycline group, the example 5 group, the comparative example 1 group, the comparative example 2 group and the comparative example 3 group of broiler chickens.

[0029] Figure 3 The treatment effective rate and mortality comparison chart of the blank control group, the model group, the florfenicol group, the doxycycline group, the example 5 group, the comparative example 1 group, the comparative example 2 group and the comparative example 3 group of broiler chickens. DETAILED DESCRIPTION

[0030] The application will be further described in conjunction with specific examples.

[0031] Example 1

[0032] A florfenicol-doxycycline compound preparation, raw materials of which include: florfenicol 5g, doxycycline hydrochloride 5g, β-cyclodextrin 20g, pullulan 8g, nano-silicon dioxide 1g, glycerol 1g, emulsifier 1g, genipin 0.1g, anhydrous glucose 20g, sodium dodecyl sulfate 0.1g, magnesium stearate 0.1g. The emulsifier is composed of Span 80 and Tween 20 with a mass ratio of 5:1.

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

[0034] S1, β-cyclodextrin and doxycycline hydrochloride are added to 100g deionized water and stirred uniformly, pullulan, nano-silicon dioxide and glycerol are added thereto and ultrasonically treated for 10min, the ultrasonic frequency being 20kHz, to obtain a first system; the emulsifier is uniformly mixed with 400g liquid paraffin to obtain a second system;

[0035] S2, the second system is continuously stirred at a stirring speed of 2000r / min, the system temperature is kept at 40℃, the first system is added dropwise thereto during the stirring process, after the dropwise addition is completed, the stirring is continued for 5min, then the pH=8.5 phosphate buffer solution containing genipin is added dropwise thereto, the stirring is carried out at 45℃ for 1.5h, the system is left to stand for 1h, the oil phase is removed by centrifugation, the product is washed with anhydrous ethanol and deionized water in sequence, and vacuum drying is carried out to obtain the doxycycline hydrochloride loaded product;

[0036] S3, florfenicol, the doxycycline hydrochloride loaded product, sodium dodecyl sulfate, magnesium stearate and 8g anhydrous glucose are uniformly mixed, and are placed in a fluidized bed as a substrate; the remaining anhydrous glucose is dissolved in 48mL purified water to prepare a binder; the binder is added to the fluidized bed for granulation, the inlet air temperature is 60℃, the fan frequency is 10Hz, the material temperature is 40℃, the liquid spraying speed is 2rpm, and the atomization pressure is 0.05MPa.

[0037] Example 2

[0038] A florfenicol-doxycycline compound preparation, raw materials of which include: florfenicol 9g, doxycycline hydrochloride 20g, beta-cyclodextrin 35g, pullulan 14g, nano-silicon dioxide 3g, glycerol 5g, emulsifier 2g, genipin 0.3g, malt dextrin 60g, sodium dodecyl sulfate 1g, talc 1g. The emulsifier is composed of Span 80 and Tween 20 in a mass ratio of 6:1.

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

[0040] S1, beta-cyclodextrin and doxycycline hydrochloride are added to 200g of deionized water and stirred uniformly, pullulan, nano-silicon dioxide and glycerol are added thereto and ultrasonically treated for 20min, the ultrasonic frequency being 25kHz, to obtain a first system; the emulsifier is uniformly mixed with 600g of liquid paraffin to obtain a second system;

[0041] S2, the second system is continuously stirred at a stirring speed of 4000r / min, the system temperature is kept at 45℃, the first system is added dropwise thereto during stirring, stirring is continued for 10min after the dropwise addition is completed, a pH=8.5 phosphate buffer solution containing genipin is added dropwise thereto, stirring is carried out at 55℃ for 2.5h, the system is left to stand for 2h, the oil phase is removed by centrifugation, the product is washed with anhydrous ethanol and deionized water in sequence, and vacuum drying is carried out to obtain doxycycline hydrochloride loaded;

[0042] S3, florfenicol, doxycycline hydrochloride loaded, sodium dodecyl sulfate, talc and 36g of malt dextrin are uniformly mixed and placed in a fluidized bed as a substrate; the remaining malt dextrin is dissolved in 144mL of purified water to obtain a binder; the binder is added to the fluidized bed for granulation, the inlet air temperature is 70℃, the fan frequency is 15Hz, the material temperature is 50℃, the liquid spraying speed is 5rpm, and the atomization pressure is 0.15MPa.

[0043] Example 3

[0044] A florfenicol-doxycycline compound preparation, raw materials of which include: florfenicol 7g, doxycycline hydrochloride 16g, hydroxypropyl-beta-cyclodextrin 25g, pullulan 12g, nano-silicon dioxide 1.5g, glycerol 4g, emulsifier 1.2g, genipin 0.25g, lactose 30g, sodium dodecyl sulfate 0.7g, magnesium stearate 0.5g. The emulsifier is composed of Span 80 and Tween 20 in a mass ratio of 5.8:1.

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

[0046] S1, hydroxypropyl-β-cyclodextrin, doxycycline hydrochloride is added to 120 g of deionized water and stirred uniformly, and then pullulan, nano-silicon dioxide and glycerol are added and ultrasonic treated for 18 min at a frequency of 21 kHz to obtain a first system; the emulsifier and 550 g of liquid paraffin are mixed uniformly to obtain a second system;

[0047] S2, the second system is continuously stirred at a speed of 2500 r / min, the temperature of the system is kept at 43°C, the first system is added dropwise during stirring, stirring is continued for 7 min after the dropwise addition is completed, and then a pH = 8.5 phosphate buffer containing genipin is added dropwise, stirring is carried out at 50°C for 100 min, and then the system is left to stand for 80 min, the oil phase is removed by centrifugation, and the product is washed with anhydrous ethanol and deionized water in sequence, and then vacuum dried to obtain doxycycline hydrochloride loaded;

[0048] S3, florfenicol, doxycycline hydrochloride loaded, lactose, sodium dodecyl sulfate and magnesium stearate are uniformly mixed, and then granulation is carried out by using a pressure roller, the feeding speed is 25 rpm, the roller speed is 8 rpm, the pressure roller pressure is 2 MPa, the pre-granulation speed is 50 rpm, the final granulation speed is 80 rpm, and 20-mesh screen granulation is carried out.

[0049] Example 4

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

[0051] The preparation method of the florfenicol-doxycycline compound preparation, which comprises the following steps:

[0052] S1, hydroxypropyl-β-cyclodextrin, doxycycline hydrochloride is added to 120 g of deionized water and stirred uniformly, and then pullulan, nano-silicon dioxide and glycerol are added and ultrasonic treated for 18 min at a frequency of 21 kHz to obtain a first system; the emulsifier and 550 g of liquid paraffin are mixed uniformly to obtain a second system;

[0053] S2, the second system is continuously stirred at a speed of 2500 r / min, the temperature of the system is kept at 43°C, the first system is added dropwise during stirring, stirring is continued for 7 min after the dropwise addition is completed, and then a pH = 8.5 phosphate buffer containing genipin is added dropwise, stirring is carried out at 50°C for 100 min, and then the system is left to stand for 80 min, the oil phase is removed by centrifugation, and the product is washed with anhydrous ethanol and deionized water in sequence, and then vacuum dried to obtain doxycycline hydrochloride loaded;

[0054] S3, uniformly mix florfenicol, loaded doxycycline hydrochloride, lactose, sodium dodecyl sulfate, talc; then use a roller to granulate, the feeding speed is 28 rpm, the roller speed is 10 rpm, the pressure of the roller is 3 MPa, the pre-granulation speed is 70 rpm, the final granulation speed is 100 rpm, and 20 mesh screen granulation is used.

[0055] Example 5

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

[0057] The preparation method of the florfenicol-doxycycline compound preparation, comprising the following steps:

[0058] S1, hydroxypropyl-β-cyclodextrin and doxycycline hydrochloride are added to 150 g of deionized water and stirred uniformly, pullulan, nano silicon dioxide and glycerol are added thereto and ultrasonically treated for 15 min at an ultrasonic frequency of 22.5 kHz to obtain a first system; an emulsifier and 500 g of liquid paraffin are uniformly mixed 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℃, drop the first system into the second system after stirring, continue to stir for 8 min, then drop the pH=8.5 phosphate buffer solution containing genipin into the system, stir at 50℃ for 120 min, stand for 90 min, remove the oil phase by centrifugation, and then wash the product with anhydrous ethanol and deionized water successively, and vacuum dry to obtain the loaded doxycycline hydrochloride;

[0060] S3, uniformly mix florfenicol, loaded doxycycline hydrochloride, lactose, sodium dodecyl sulfate and magnesium stearate; then use a roller to granulate, the feeding speed is 25 rpm, the roller speed is 8 rpm, the pressure of the roller is 2.5 MPa, the pre-granulation speed is 50 rpm, the final granulation speed is 80 rpm, and 20 mesh screen granulation is used.

[0061] Comparative Example 1

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

[0063] The preparation method of the florfenicol-doxycycline compound preparation includes the following steps:

[0064] The florfenicol, the doxycycline hydrochloride, the hydroxypropyl-beta-cyclodextrin, the lactose, the sodium dodecyl sulfate and the magnesium stearate are uniformly mixed, and then granulation is carried out by using a roller, the feeding rotation speed is 25 rpm, the roller rotation speed is 8 rpm, the roller pressure is 2.5 MPa, the pre-granulation rotation speed is 50 rpm, the final granulation rotation speed is 80 rpm, and 20-mesh screen granulation is carried out.

[0065] Comparative Example 2

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

[0067] The preparation method of the florfenicol-doxycycline compound preparation includes the following steps:

[0068] S1, the hydroxypropyl-beta-cyclodextrin and the doxycycline hydrochloride are added to 150 g of deionized water and stirred uniformly, nano-silicon dioxide and glycerol are added thereto and ultrasonically treated for 15 min, the ultrasonic frequency is 22.5 kHz, to obtain a first system; the emulsifier and 500 g of liquid paraffin are uniformly mixed to obtain a second system;

[0069] S2, the second system is continuously stirred at a stirring speed of 3000 r / min, the system temperature is kept at 42℃, the first system is added dropwise into the second system during the stirring process, after the dropwise addition is completed, the stirring is continued for 8 min, and the product is washed with anhydrous ethanol and deionized water in sequence, and vacuum dried to obtain the doxycycline hydrochloride loaded product;

[0070] S3, the florfenicol, the doxycycline hydrochloride loaded product, the lactose, the sodium dodecyl sulfate and the magnesium stearate are uniformly mixed, and then granulation is carried out by using a roller, the feeding rotation speed is 25 rpm, the roller rotation speed is 8 rpm, the roller pressure is 2.5 MPa, the pre-granulation rotation speed is 50 rpm, the final granulation rotation speed is 80 rpm, and 20-mesh screen granulation is carried out.

[0071] Comparative Example 3

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

[0073] The preparation method of the florfenicol-doxycycline compound preparation, comprising the following steps:

[0074] S1, hydroxypropyl-beta-cyclodextrin and doxycycline hydrochloride are added to 150 g of deionized water and stirred uniformly, and pullulan, nano-silicon dioxide, and glycerol are added thereto and ultrasonically treated for 15 min at an ultrasonic frequency of 22.5 kHz to obtain a first system;

[0075] S2, pH = 8.5 phosphate buffer containing genipin is added dropwise to the first system, and stirred at 50 ℃ for 120 min, and then left to stand for 90 min, and the product is sequentially washed with anhydrous ethanol and deionized water, and vacuum dried to obtain doxycycline hydrochloride loaded;

[0076] S3, florfenicol, doxycycline hydrochloride loaded, lactose, sodium dodecyl sulfate, and magnesium stearate are uniformly mixed, and then granulated by using a compression roller, with a feeding rotation speed of 25 rpm, a compression roller rotation speed of 8 rpm, a compression wheel pressure of 2.5 MPa, a pre-granulation rotation speed of 50 rpm, a final granulation rotation speed of 80 rpm, and a 20-mesh screen.

[0077] 1.0 g of the preparation samples obtained in Example 5 and Comparative Examples 1-3 is placed in a beaker containing 100 mL of purified water at 25 ℃, and it is observed that the preparation sample is completely dissolved after stirring at 400 rpm for 10 min, and the solution is clear.

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

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

[0080]

[0081]

[0082] According to the above results, it can be seen 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 ℃ and a relative humidity of 75±5%.

[0083] The formulation samples obtained in Example 5 and Comparative Examples 1-3 were placed in a constant temperature and humidity chamber at 60±2℃ for 10 days. Samples were taken on day 0, day 5 and day 10 to test the properties and content of the samples. The results are shown in Table 2.

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

[0085]

[0086] Based on the above results, it can be seen that the formulation sample obtained in Example 5 has better stability than Comparative Examples 1-3 in a high-temperature environment of 60±2℃ (P<0.05).

[0087] The formulation samples obtained in Example 5 and Comparative Examples 1-3 were placed in a constant temperature and humidity chamber at room temperature and relative humidity of 90±5% for 10 days. Samples were taken on day 0, day 5 and day 10 to test the properties and content of the samples. The results are shown in Table 3.

[0088] Table 3. High-humidity stability test of florfenicol-doxycycline combination preparations

[0089]

[0090] Based on the above results, it can be seen that the formulation sample obtained in Example 5 has the best stability 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 (MICs) (μg / mL) of the formulations 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 ATCC11859 were determined using the micro-broth dilution method.

[0092] Experimental results are as follows Figure 1 As shown, the minimum inhibitory concentrations (MICs) of the formulations obtained in Example 5 and Comparative Examples 1-3 were all lower than those of 10% florfenicol powder and 10% doxycycline hydrochloride soluble powder; while the MICs of Comparative Example 1 were the lowest in all categories. This should be because Example 5 and Comparative Examples 2-3 used more excipients than Comparative Example 1; however, the MIC of the formulation obtained in Example 5 was lower than that of Comparative Examples 2-3, and the antibacterial effect was 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 weight were taken, and were allowed to freely eat and drink water for 1 week, and were randomly divided into 8 groups, 15 broilers in each group, which were 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. Except for the blank control group, the rest of the broilers were injected with 0.3 mL of 1×10 9 CFU / mL of E. coli into the abdominal cavity to establish a chicken E. coli disease model.

[0094] All groups were bred according to the conventional feeding method, wherein: the florfenicol group was administered with 10% florfenicol powder (mixed with water, 1 g of 10% florfenicol powder was added to 1 L of water), the doxycycline group was administered with 10% doxycycline hydrochloride soluble powder (mixed with water, 6 g of 10% doxycycline hydrochloride soluble powder was added to 1 L of water), the Example 5 group was administered with the preparation obtained in Example 5 (mixed with water, 3.5 g of the preparation obtained in Example 5 was added to 1 L of water), the Comparative Example 1 group was administered with the preparation obtained in Comparative Example 1 (mixed with water, 3.5 g of the preparation obtained in Comparative Example 1 was added to 1 L of water), the Comparative Example 2 group was administered with the preparation obtained in Comparative Example 2 (mixed with water, 3.5 g of the preparation obtained in Comparative Example 2 was added to 1 L of water), and the Comparative Example 3 group was administered with the preparation obtained in Comparative Example 3 (mixed with water, 3.5 g of the preparation obtained in Comparative Example 3 was added to 1 L of water). The administration was continuously performed for 7 days. The broilers in each group were weighed, and the treatment efficiency and mortality rate were calculated.

[0095] As shown in Table 1, the broilers in the Example 5 group had the highest body weight, which was better than that of the broilers in the florfenicol group, the doxycycline group and the Comparative Example 1-3 groups (P<0.05), and was close to that of the broilers in the blank control group, and the two groups lacked significant difference (P>0.05). Figure 2 As shown in Table 2, the treatment efficiency of the broilers in the Example 5 group and the Comparative Example 3 group was 100%, which was better than that of the broilers in the other groups (P<0.05); and the mortality rate of the broilers in the Example 5 group and the Comparative Example 1-3 groups was 0, which was better than that of the broilers in the model group, the florfenicol group and the doxycycline group (P<0.05).

[0096] Figure 3 As shown in Table 2, the treatment efficiency of the broilers in the Example 5 group and the Comparative Example 3 group was 100%, which was better than that of the broilers in the other groups (P<0.05); and the mortality rate of the broilers in the Example 5 group and the Comparative Example 1-3 groups was 0, which was better than that of the broilers in the model group, the florfenicol group and the doxycycline group (P<0.05).

[0097] The applicant believes that: this is because the high efficient loading of doxycycline hydrochloride is realized by the synergistic effect of the inclusion of cyclodextrin and the embedding of polyacrylamide network, the product is porous microparticles with potential for sustained release, and the loading of doxycycline hydrochloride and florfenicol is compounded, which not only has excellent stability of the preparation, but also greatly enhances the stability in high temperature and high humidity. The water-insoluble florfenicol and strong polar doxycycline hydrochloride are loaded, which not only overcomes the shortcoming that water-soluble and fat-soluble drugs are difficult to blend, but also reduces the minimum inhibitory concentration of E. coli, Staphylococcus aureus and Pasteurella multocida, prolongs the action time of the two in the animal body, maintains stable synergistic antibacterial effect in the body, and has remarkable clinical treatment effect. ​

[0098] The above merely provides the preferred but not limiting embodiments of the present application, and the protection scope of the present application should not be limited thereto. Any skilled person in the art, according to the technical solution and the inventive concept of the present application, can make equivalent replacement or change within the technical range disclosed by the present application, which should be covered in the protection scope of the present application.

Claims

1. A florfenicol-doxycycline compound preparation, characterized in that, The raw materials, by weight, include: 5-10 parts florfenicol, 5-20 parts doxycycline hydrochloride, 20-40 parts inclusion material, 8-14 parts pullulan polysaccharide, 1-3 parts nano silica, 1-5 parts glycerin, 1-2 parts emulsifier, 0.1-0.3 parts genipin, 20-60 parts filler, 0.1-1 part surfactant, and 0.1-1 part lubricant; The inclusion material is β-cyclodextrin and / or hydroxypropyl-β-cyclodextrin; The following steps are used to prepare it: S1. Add the inclusion material and doxycycline hydrochloride to water and stir until homogeneous. Add pullulan polysaccharide, nano-silica, and glycerin to the mixture and sonicate for 10-20 minutes to obtain the first system. Mix the emulsifier and liquid paraffin evenly to obtain the second system. S2. Continue stirring the second system while maintaining the system temperature at 40-45℃. During the stirring process, add the first system dropwise. After the addition is complete, continue stirring for 5-10 minutes. Then, add phosphate buffer containing genipin dropwise. Stir at 45-55℃ for 1.5-2.5 hours, let stand for 1-2 hours, centrifuge to remove the oil phase, wash, and vacuum dry to obtain doxycycline hydrochloride-loaded. S3 is granulated using florfenicol, doxycycline hydrochloride, fillers, surfactants, and lubricants.

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

3. The florfenicol-doxycycline compound preparation according to claim 1, characterized in that, The surfactant is sodium dodecyl sulfate.

4. The florfenicol-doxycycline compound preparation according to claim 1, characterized in that, The lubricant is talc and / or magnesium stearate.

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

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

1.

7. A method for preparing a florfenicol-doxycycline compound preparation as described in any one of claims 1-6, characterized in that, Includes the following steps: S1. Add the inclusion material and doxycycline hydrochloride to water and stir until homogeneous. Add pullulan polysaccharide, nano-silica, and glycerin to the mixture and sonicate for 10-20 minutes to obtain the first system. Mix the emulsifier and liquid paraffin evenly to obtain the second system. S2. Continue stirring the second system while maintaining the system temperature at 40-45℃. During the stirring process, add the first system dropwise. After the addition is complete, continue stirring for 5-10 minutes. Then, add phosphate buffer containing genipin dropwise. Stir at 45-55℃ for 1.5-2.5 hours, let stand for 1-2 hours, centrifuge to remove the oil phase, wash, and vacuum dry to obtain doxycycline hydrochloride-loaded. S3 is granulated using florfenicol, doxycycline hydrochloride, fillers, surfactants, and lubricants.

8. The method for preparing the florfenicol-doxycycline compound preparation according to claim 7, characterized in that, In S1, the ultrasonic frequency is 20-25kHz.

9. The method for preparing the florfenicol-doxycycline compound preparation according to claim 7, characterized in that, In S2, the pH value of the phosphate buffer is 8.5.

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