Ceftiofur hydrochloride injection and preparation method thereof

Abstract

The invention discloses a ceftiofur hydrochloride injection, which is prepared from the following components in parts by weight: 5 to 11 parts of ceftiofur hydrochloride, 1 to 2 parts of suspending emulsifier, 0.01 to 0.04 part of antioxidant and 45 to 66 parts of oil solvent, the oil solvent is one or more of caprylic capric triglyceride, refined coconut oil and cottonseed oil; the suspending emulsifier is lipophilic sucrose fatty acid ester and / or aluminum stearate. The preparation is advanced and reasonable in prescription design, cheap and easily available in raw materials and auxiliary materials, stable and reliable in preparation process, simple and easy to operate, good in stability, easy to suck and inject, low in irritation of an injection part, good in absorption and high in drug bioavailability, can improve animal compliance and enhance the disease treatment effect, and can also be used for preparing the medicine for treating the diseases. The risk that the drug resistance of bacteria is increased due to repeated administration and mixed administration caused by poor curative effect of the preparation can be reduced, and the treatment cost is reduced.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations and relates to a ceftiofur hydrochloride injection and a preparation method thereof. Background Art

[0002] Ceftiofur is a third-generation cephalosporin antibiotic for animals. It has a broad antibacterial spectrum and strong antibacterial activity. It is effective against Gram-negative bacteria such as Escherichia coli, Salmonella, Pasteurella multocida and hemolytic Pasteurella, Actinobacillus pleuropneumoniae, etc. After intramuscular injection, the drug is rapidly absorbed, widely distributed, has a long-lasting efficacy, low toxicity, and few side effects. It is often used to treat and control animal respiratory and urinary tract infections caused by sensitive bacteria, and has broad application prospects in veterinary clinical treatment of livestock and poultry diseases.

[0003] Currently, the main formulations of ceftriaxone on the market are powder injection and injection solution. Among them, most powder injections are directly packaged from raw materials or made into freeze-dried powder injections. Such preparations need to be dissolved in physiological saline before use before injection, and the storage conditions are relatively strict. Improper storage can easily cause the preparation to be unstable, resulting in drug failure and affecting the treatment effect. In addition, such preparations have no protection and "assistance" from excipients, so the local irritation of the drug to the injection site and the problem of sustained release of the drug cannot be well solved, which limits its application.

[0004] Ceftiofur hydrochloride injection belongs to an oil suspension preparation, which has the characteristics of easy use and long-acting sustained release, makes up for the shortcomings of the above-mentioned powder injection for injection, and is favored by the majority of users. However, the existing ceftiofur hydrochloride injection still has unreasonable parts in the prescription design, the obtained suspension particles are too large or too fine, the fluidity is poor, the redispersion is poor, the stability is poor, and the defects such as too fast sedimentation, needle blockage, difficulty in suction, difficulty in push injection, poor absorption at the injection site, and large local irritation occur during clinical use. For example, Chinese patent 201010549740.2 discloses a long-acting ceftiofur hydrochloride injection, which adopts a composite solvent containing polyethylene glycol, a suspending agent is a complex of hydrogenated castor oil and lecithin, and an antibacterial agent is chlorobutanol or benzyl alcohol, and is prepared by high-speed shearing disperser and colloid mill dispersion. However, polyethylene glycol has a slight solubility effect on ceftiofur hydrochloride, and hydrogenated castor oil is easy to harden at the bottom, affecting redispersibility, especially when the transportation and storage conditions are not guaranteed during market circulation. It is very easy for the quality of suspension preparations to decline due to excipient problems, affecting disease treatment. Chinese patent 201710323608.1 discloses a ceftiofur hydrochloride injection for dairy cows, the components of which include ceftiofur hydrochloride, suspending agents, flocculants, emulsifiers, antioxidants and lipid solvents. However, as an oil suspension preparation, adding a flocculant is actually not advisable. First, the flocculant is mainly used in water suspensions to adjust stability and dispersibility. Second, the flocculant is mainly related to pH. The pH of the oil suspension cannot be measured. Adding a flocculant is meaningless and increases quality risks. In addition, suspending agents such as water-soluble suspending agents such as chitosan, sodium carboxymethyl cellulose, hydroxypropyl methylcellulose, gelatin, etc., cannot play a suspending effect in oil; emulsifiers such as hydrogenated castor oil and soybean lecithin are prone to bottom condensation over time after homogenization and emulsification, resulting in poor redispersion; lipid solvents such as soybean oil and white oil have a lower viscosity after mixing, but white oil is highly irritating and increases irritation at the injection site. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a ceftiofur hydrochloride injection and a preparation method thereof, which has advanced and reasonable prescription design and stable and reliable preparation process, and can not only improve the quality of drug preparations, increase drug utilization, and enhance disease treatment effects, but also reduce the risk of increased bacterial resistance due to multiple administration and mixed medication caused by poor efficacy of drug preparations, while reducing treatment costs.

[0006] After research, the present invention provides the following technical solutions:

[0007] 1. Ceftiofur hydrochloride injection is composed of the following components by weight: 5-11 parts of ceftiofur hydrochloride, 1-2 parts of suspending emulsifier, 0.01-0.04 parts of antioxidant, and 45-66 parts of oil solvent; the suspending emulsifier is lipophilic sucrose fatty acid ester and / or aluminum stearate; the oil solvent is one or more of caprylic decanoic acid glyceride, refined coconut oil and cottonseed oil.

[0008] Preferably, the ceftiofur hydrochloride injection consists of the following components by weight: 5-11 parts of ceftiofur hydrochloride, 1-2 parts of suspending emulsifier, 0.01-0.04 parts of antioxidant, and 60-65 parts of oil solvent; the suspending emulsifier is lipophilic sucrose fatty acid ester and / or aluminum stearate; the oil solvent is caprylic decanoic acid glyceride and / or refined coconut oil.

[0009] As one of the preferred technical solutions, the ceftiofur hydrochloride injection is composed of the following components by weight: 5-6 parts of ceftiofur hydrochloride, 2 parts of aluminum stearate, 0.02 parts of antioxidant BHA, and 64-65 parts of caprylic decanoate.

[0010] As one of the preferred technical solutions, the ceftiofur hydrochloride injection is composed of the following components by weight: 5-6 parts of ceftiofur hydrochloride, 1.5 parts of aluminum stearate, 0.5 parts of lipophilic sucrose fatty acid ester, 0.02 parts of antioxidant BHA, and 64-65 parts of caprylic decanoic acid glyceride.

[0011] As one of the preferred technical solutions, the ceftiofur hydrochloride injection is composed of the following components by weight: 10-11 parts of ceftiofur hydrochloride, 1.5 parts of aluminum stearate, 0.04 parts of antioxidant BHA, and 61-62 parts of caprylic decanoate.

[0012] As one of the preferred technical solutions, the ceftiofur hydrochloride injection is composed of the following components by weight: 5-6 parts of ceftiofur hydrochloride, 2 parts of lipophilic sucrose fatty acid ester, 0.02 parts of antioxidant BHA, and 61-62 parts of caprylic decanoic acid glyceride.

[0013] As the best technical solution, the ceftiofur hydrochloride injection is composed of the following components by weight: 5-6 parts of ceftiofur hydrochloride, 1.2 parts of aluminum stearate, 0.02 parts of antioxidant BHA, 28-29 parts of refined coconut oil, and 35-36 parts of caprylic decanoic acid glyceride.

[0014] 2. The preparation method of the ceftiofur hydrochloride injection comprises the following steps: taking ceftiofur hydrochloride, dispersing it in part of the oil solvent to obtain a crude ceftiofur hydrochloride solution; taking a suspending emulsifier, dispersing it in the remaining oil solvent, heating it to 75-125°C, stopping heating, waiting to cool to 50-65°C, adding an antioxidant, stirring to completely dissolve, waiting to cool to 35-45°C, adding the crude ceftiofur hydrochloride solution, stirring evenly, waiting to cool to 25-35°C, transferring it into a high-pressure homogenizer, first circulating homogenization at a pressure of 5-20Mpa for 10-30min, and then repeating the homogenization twice at a pressure of 10-30Mpa to obtain ceftiofur hydrochloride injection.

[0015] The preparation of the ceftiofur hydrochloride injection of the present invention uses an oil solvent as a dispersion medium and uses high pressure conditions to homogenize and disperse the drug.

[0016] Preferably, the preparation method of the ceftiofur hydrochloride injection comprises the following steps: taking ceftiofur hydrochloride, dispersing it in part of the oil solvent to obtain a crude ceftiofur hydrochloride solution; taking a suspending emulsifier, dispersing it in the remaining oil solvent, heating it to 75-120°C, stopping heating, waiting to cool to 55-60°C, adding an antioxidant, stirring to completely dissolve, waiting to cool to 35°C, adding the crude ceftiofur hydrochloride solution, stirring for 15-20 minutes, waiting to cool to 30°C, transferring it into a high-pressure homogenizer, first circulating homogenization at a pressure of 8-15Mpa for 15-30 minutes, and then repeating the homogenization twice at a pressure of 15-25Mpa to obtain ceftiofur hydrochloride injection.

[0017] The beneficial effects of the present invention are as follows: the present invention provides a ceftiofur hydrochloride injection by strictly screening the auxiliary material components of the preparation prescription and optimizing the preparation process; the prescription design is advanced and reasonable; the raw and auxiliary materials are cheap and easily available; the preparation process is stable and reliable, simple and easy to operate; the obtained preparation has good stability, is easy to aspirate and push, has low irritation at the injection site, is well absorbed, and has high drug bioavailability; it can not only improve animal compliance and enhance the effect of disease treatment, but also reduce the risk of increased bacterial resistance due to multiple administration and mixed medication caused by poor preparation efficacy, thereby reducing treatment costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is a comparison diagram of the drug-time curves of the ceftiofur hydrochloride injection prepared by the present invention and the imported commercially available preparation RLD. DETAILED DESCRIPTION

[0019] In order to make the objectives, technical solutions and beneficial effects of the present invention more clear, the preferred embodiments of the present invention are described in detail below.

[0020] Example 1 Screening of oil solvent

[0021] 1 Materials and methods

[0022] 1.1 Test samples: soybean oil, white oil for injection, caprylic / capric glyceride, refined coconut oil, ethyl oleate, cottonseed oil, propylene glycol dicaprylate / caprate, and polydimethylsiloxane.

[0023] 1.2 Methods

[0024] One week before the experiment, observe whether the rabbits' spirit, activity, appetite, body temperature, etc. are normal. During the experiment, take 4 clean New Zealand white rabbits, use the self-left and right control method, cut the hair of the hind limbs, disinfect the rectus femoris of the left and right quadriceps with alcohol cotton balls, vertically insert the needle at the 1 / 2 between the front of the thigh, the knee joint and the femoral joint, slowly inject (1mL / 10s) 1mL of the test sample on the right side, and inject the same dose of normal saline at the same site on the left side. After the injection, quickly remove the needle without massaging the injection site. Check whether there are any changes in the injection site 48h after administration. After 72h, the rabbits were killed, the quadriceps were removed by dissection, cut longitudinally, and the local irritation reaction of the injection was observed with the naked eye. The reaction level was obtained according to the muscle irritation test reaction scoring standard in Table 1, and the average reaction level was obtained. When the average reaction level is below 2, it can be used for intramuscular injection; when the average reaction level exceeds 3, it cannot be used for intramuscular injection; when the average reaction level is between 2 and 3, repeated tests can be carried out or combined with other indicators for comprehensive consideration.

[0025] Table 1 Scoring criteria for muscle stimulation test reaction

[0026]

[0027]

[0028] 2 Test results

[0029] The results of rabbit muscle stimulation test with different oil solvents are shown in Table 2. There is no significant change in the administration site of caprylic decanoic acid glyceride, cottonseed oil, refined coconut oil, and polydimethicone, but polydimethicone is not absorbed by the body, so the oil solvent is preferably one or more of caprylic decanoic acid glyceride, cottonseed oil, and refined coconut oil.

[0030] Table 2 Scoring results of rabbit muscle irritation test with different oil solvents

[0031]

[0032] Example 2 Screening of suspending emulsifiers

[0033] 1 Materials and methods

[0034] 1.1 Test samples: aluminum stearate, hydrogenated castor oil, lecithin, lipophilic sucrose fatty acid esters, hydrogenated lecithin, nano-silicon dioxide, sorbitan oleate, polyoxyethylene sorbitan monooleate.

[0035] 1.2 Methods

[0036] Weigh 1.25g of aluminum stearate, disperse it in 45.82g of caprylic acid glyceride, heat it to 118°C, and stop heating; take another 7 portions of 45.82g of caprylic acid glyceride and heat it to 80°C, add 0.75% of hydrogenated castor oil, lecithin, lipophilic sucrose fatty acid ester, hydrogenated lecithin, nano-silicon dioxide, sorbitan oleate, polyoxyethylene sorbitan monooleate, stir to dissolve completely, and stop heating; wait for the above 8 mixed solutions to cool to 55°C, add antioxidants respectively, stir to dissolve completely, and wait for cooling. To 35°C, add crude ceftiofur hydrochloride solution (weigh 8 parts of 5.45g ceftiofur hydrochloride and disperse them in 28.65g refined coconut oil respectively), stir for 30min, wait until cooled to 30°C, transfer to a high-pressure homogenizer, adjust the pressure to 8Mpa and circulate homogenization for 20min, then adjust the pressure to 20Mpa for the first homogenization, and after the end, adjust the pressure to 15Mpa for the second homogenization to obtain preparations (ceftiofur hydrochloride injection) A1, A2, A3, A4, A5, A6, A7, A8, respectively.

[0037] The obtained preparations A1 to A8 were examined for dispersibility, sedimentation volume ratio, needle extraction properties and appearance properties as follows.

[0038] Dispersibility: Take one bottle of the preparation, shake vigorously for 30 seconds, and transfer to a glass container. No lumps or sediments should be observed.

[0039] Sedimentation volume ratio: take 1 bottle of preparation, shake for 1 minute, take 50 ml with a stoppered measuring cylinder, stopper it tightly, shake it vigorously for 1 minute, record the initial height H0 of the suspension, let it stand for 3 hours, and record the final height H of the suspension. The sedimentation volume ratio (H / H0) should not be less than 0.90.

[0040] Needle withdrawal test: Take 1 bottle of preparation, shake it evenly, and use a 5ml syringe equipped with a No. 7 needle to inject 5ml of air. More than 1ml of the content should be able to be smoothly withdrawn within 3 minutes.

[0041] Appearance properties: Observe at room temperature and conduct accelerated test at 60℃ for 10 days.

[0042] 2 Test results

[0043] The effects of different suspending emulsifiers on some indicators of ceftiofur hydrochloride injection are shown in Table 3. Different suspending emulsifiers have a certain effect on the dispersibility, sedimentation volume ratio, needle extraction and appearance of ceftiofur hydrochloride injection. Among them, aluminum stearate and lipophilic sucrose fatty acid ester have good comprehensive performance and can be used alone or in combination.

[0044] Table 3 Effects of different suspending emulsifiers on ceftiofur hydrochloride injection

[0045]

[0046] Example 3 Preparation of Ceftiofur Hydrochloride Injection (Preparation B1)

[0047] Weigh 5.45 g of ceftiofur hydrochloride, disperse it in 28.65 g of caprylic acid glyceride to obtain a crude ceftiofur hydrochloride solution; weigh 2.0 g of aluminum stearate, disperse it in 35.82 g of caprylic acid glyceride, heat to 118° C., stop heating, wait until cooled to 55° C., add 0.02 g of antioxidant BHA, stir to dissolve completely, wait until cooled to 35° C., add the above-mentioned crude ceftiofur hydrochloride solution, stir for 20 minutes, wait until cooled to 30° C., transfer it into a high-pressure homogenizer, adjust the pressure to 10 MPa, circulate homogenization for 15 minutes, then adjust the pressure to 25 MPa for the first homogenization, and after the end, adjust the pressure to 20 MPa for the second homogenization to obtain Preparation B1.

[0048] Example 4 Preparation of Ceftiofur Hydrochloride Injection (Preparation B2)

[0049] Weigh 5.45g of ceftiofur hydrochloride, disperse it in 35.75g of caprylic acid glyceride to obtain a crude ceftiofur hydrochloride solution; weigh 1.5g of aluminum stearate, disperse it in 28.54g of caprylic acid glyceride, heat it to 118°C, stop heating, wait until it cools to 75°C, add 0.5g of lipophilic sucrose fatty acid ester, wait until it cools to 60°C, add 0.02g of antioxidant BHA, stir to dissolve completely, wait until it cools to 35°C, add the above-mentioned crude ceftiofur hydrochloride solution, stir for 15min, wait until it cools to 30°C, transfer it into a high-pressure homogenizer, adjust the pressure to 12Mpa, circulate and homogenize for 20min, then adjust the pressure to 20Mpa for the first homogenization, and after the end, adjust the pressure to 15Mpa for the second homogenization to obtain Preparation B2.

[0050] Example 5 Preparation of Ceftiofur Hydrochloride Injection (Preparation B3)

[0051] Weigh 10.90g of ceftiofur hydrochloride, disperse it in 25.65g of caprylic acid glyceride to obtain a crude ceftiofur hydrochloride solution; weigh 1.5g of aluminum stearate, disperse it in 35.82g of caprylic acid glyceride, heat to 118°C, stop heating, wait until cooled to 55°C, add 0.04g of antioxidant BHA, stir to dissolve completely, wait until cooled to 35°C, add the above-mentioned crude ceftiofur hydrochloride solution, stir for 20min, wait until cooled to 30°C, transfer it into a high-pressure homogenizer, adjust the pressure to 10Mpa and circulate homogenization for 30min, then adjust the pressure to 25Mpa for the first homogenization, and after the end, adjust the pressure to 25Mpa for the second homogenization to obtain Preparation B3.

[0052] Example 6 Preparation of Ceftiofur Hydrochloride Injection (Preparation B4)

[0053] Weigh 10.90g of ceftiofur hydrochloride, disperse it in 25.65g of caprylic acid glyceride to obtain a crude ceftiofur hydrochloride solution; weigh 1.5g of aluminum stearate, disperse it in 35.82g of caprylic acid glyceride, heat it to 118°C, stop heating, wait until it cools to 75°C, add 0.5g of lipophilic sucrose fatty acid ester, wait until it cools to 55°C, add 0.04g of antioxidant BHA, stir to dissolve completely, wait until it cools to 35°C, add the above-mentioned crude ceftiofur hydrochloride solution, stir for 15min, wait until it cools to 30°C, transfer it into a high-pressure homogenizer, adjust the pressure to 10Mpa and circulate homogenization for 20min, then adjust the pressure to 20Mpa for the first homogenization, and after the end, adjust the pressure to 18Mpa for the second homogenization to obtain Preparation B4.

[0054] Example 7 Preparation of Ceftiofur Hydrochloride Injection (Preparation B5)

[0055] Weigh 10.90g of ceftiofur hydrochloride, disperse it in 25.65g of caprylic acid glyceride to obtain a crude ceftiofur hydrochloride solution; weigh 2.0g of lipophilic sucrose fatty acid ester, disperse it in 35.82g of caprylic acid glyceride, heat to 78°C, stop heating, wait until cooled to 55°C, add 0.04g of antioxidant BHA, stir to dissolve completely, wait until cooled to 35°C, add the above-mentioned crude ceftiofur hydrochloride solution, stir for 15min, wait until cooled to 30°C, transfer it into a high-pressure homogenizer, adjust the pressure to 15Mpa, circulate homogenization for 20min, adjust the pressure to 25Mpa for the first homogenization, and after the end, adjust the pressure to 18Mpa for the second homogenization to obtain Preparation B5.

[0056] Example 8 Preparation of Ceftiofur Hydrochloride Injection (Preparation B6)

[0057] Weigh 5.45g of ceftiofur hydrochloride, disperse it in 25.65g of caprylic acid glyceride to obtain a crude ceftiofur hydrochloride solution; weigh 2.0g of lipophilic sucrose fatty acid ester, disperse it in 35.82g of caprylic acid glyceride, heat to 78°C, stop heating, wait until cooled to 55°C, add 0.02g of antioxidant BHA, stir to dissolve completely, wait until cooled to 35°C, add the above-mentioned crude ceftiofur hydrochloride solution, stir for 15 minutes, wait until cooled to 30°C, transfer it into a high-pressure homogenizer, adjust the pressure to 8Mpa, circulate homogenization for 15 minutes, then adjust the pressure to 20Mpa for the first homogenization, and after the end, adjust the pressure to 18Mpa for the second homogenization to obtain Preparation B6.

[0058] Example 9 Preparation of Ceftiofur Hydrochloride Injection (Preparation B7)

[0059] Weigh 5.45 g of ceftiofur hydrochloride, disperse it in 10.65 g of refined coconut oil to obtain a crude ceftiofur hydrochloride solution; weigh 2.0 g of lipophilic sucrose fatty acid ester, disperse it in 35.82 g of caprylic decanoic acid glyceride, heat to 78°C, stop heating, wait until cooled to 55°C, add 0.02 g of antioxidant BHA, stir to dissolve completely, wait until cooled to 35°C, add the above-mentioned crude ceftiofur hydrochloride solution, stir for 15 minutes, wait until cooled to 30°C, transfer it into a high-pressure homogenizer, adjust the pressure to 12 MPa, circulate homogenization for 15 minutes, adjust the pressure to 15 MPa for the first homogenization, and after the end, adjust the pressure to 18 MPa for the second homogenization to obtain Preparation B7.

[0060] Example 10 Preparation of Ceftiofur Hydrochloride Injection (Preparation B8)

[0061] Weigh 5.45 g of ceftiofur hydrochloride, disperse it in 28.65 g of refined coconut oil to obtain a crude ceftiofur hydrochloride solution; weigh 1.2 g of aluminum stearate, disperse it in 35.82 g of caprylic acid glyceride, heat to 118° C., stop heating, wait until cooled to 55° C., add 0.02 g of antioxidant BHA, stir to dissolve completely, wait until cooled to 35° C., add the above-mentioned crude ceftiofur hydrochloride solution, stir for 20 minutes, wait until cooled to 30° C., transfer it into a high-pressure homogenizer, adjust the pressure to 8 MPa, circulate homogenization for 15 minutes, then adjust the pressure to 20 MPa for the first homogenization, and after the end, adjust the pressure to 18 MPa for the second homogenization to obtain preparation B8.

[0062] Example 11 Study on the stability of ceftiofur hydrochloride injection

[0063] According to the "Guidelines for Stability Testing of Veterinary Drugs", preparations B1 to B8 prepared in Examples 3 to 10 were placed in a stability test chamber at a temperature of 40±2°C and a relative humidity of 75±5% for 6 months. Samples were taken at the end of the 1st, 2nd, 3rd and 6th month to detect the content of the preparations and observe their appearance, properties and dispersibility.

[0064] The results of the stability study are shown in Tables 4 to 5. After being placed at 40±2°C and 75±5% for 6 months, the content, appearance and dispersibility of preparations B1 to B8 did not change significantly. Among them, preparation B8 prepared in Example 10 performed best in stability, and other preparations B1, B2, B3, and B6 also performed well. For this reason, other indicators of preparations B1, B2, B3, B6, and B8 were further investigated. The results are shown in Table 6. Preparation B8 prepared in Example 10 still performed best.

[0065] Table 4 Changes in the content of Ceftiofur Hydrochloride Injection

[0066]

[0067]

[0068] Table 5 Appearance and dispersibility changes of Ceftiofur Hydrochloride Injection (6th month)

[0069]

[0070] Table 6 Other evaluation indicators of ceftiofur hydrochloride injection

[0071]

[0072] Example 12 Investigation of the irritation of ceftiofur hydrochloride injection

[0073] Preparation B8 prepared in Example 10 was used for rabbit muscle irritation test according to the method described in Example 1, and physiological saline was set as a negative control. The reaction level was obtained according to the muscle irritation reaction grading standard shown in Table 1, and the average reaction level was calculated. The results showed that there was no significant difference between the preparation B8 group and the physiological saline group, and there was no irritation to the local muscles of rabbits, and the average reaction level was 0, indicating that the ceftiofur hydrochloride injection prepared by the present invention is low irritant and can be used for intramuscular injection.

[0074] Example 13 Bioequivalence Study of Ceftiofur Hydrochloride Injection

[0075] Twenty-four healthy crossbred pigs (30.38±4.12kg) were randomly divided into two groups, A and B, according to a 2×2 crossover design, with 12 pigs in each group. After 7 days of conventional feeding and observation, the reference drug was injected into the deep neck muscle of group A, and the test drug was injected into the deep neck muscle of group B, with a dose of 5 mg kg -1 After an interval of 7 days, the test drug was injected intramuscularly in group A and the reference drug was injected intramuscularly in group B, with a dose of 5 mg kg -1 (body weight). The reference drug is the imported commercial preparation RLD of ceftiofur hydrochloride injection; the test drug is the preparation B8 prepared in Example 10. An appropriate amount of blank plasma is collected before administration; 3-4 mL of blood is collected from the anterior vena cava at 0.2, 0.5, 1, 2, 4, 5, 8, 12, 24, 48, 72 and 96 h after administration, respectively, and placed in a centrifuge tube with heparin anticoagulation, centrifuged at 4000 r / min for 15 min, 0.5 ml of plasma sample is taken and added with 7 ml of DTE extraction reagent, and the mixture is placed in a 50°C water bath for 15 min (vortexed once every 5 min), and centrifuged at 8000 r / min for 10 min. After cooling to room temperature, the supernatant is taken, which is the extract.

[0076] The HLB solid phase extraction column was activated with 3 ml of methanol and 3 ml of water, and then the extract was loaded onto the column, rinsed with 2 ml of water, eluted with 3 ml of methanol, and the eluate was collected and nitrogen blown at 35°C. Finally, it was reconstituted with water to 0.5 ml and passed through a 0.22 μm organic filter membrane. 100 μL of the filtrate was injected into a high performance liquid chromatograph for determination, and the blood drug concentration at each time point was calculated. The pharmacokinetic parameters of each group were calculated using 3p97 software to evaluate the bioequivalence of the test drug and the reference drug. Chromatographic conditions: The chromatographic column was Waters C18 (4.6 mm × 250 mm, 5 μm); the mobile phase was 0.1% trifluoroacetic acid-acetonitrile = 85:15 (volume ratio); the detection wavelength was 265 nm; the flow rate was 1 mL / min; the column temperature was 35°C; and the injection volume was 100 μL.

[0077] The pharmacokinetic parameters of the two preparations after intramuscular injection into pigs (n = 24) are shown in Table 7, and the drug-time curves are shown in Figure 1 The experimental results showed that the relative bioavailability of the preparation B8 prepared in Example 10 and the imported commercial preparation RLD was 102.2%, indicating that the ceftiofur hydrochloride injection prepared in the present invention and the imported commercial preparation RLD were bioequivalent.

[0078] Table 7 Pharmacokinetic parameters of the two preparations after intramuscular injection into pigs (n=24)

[0079]

[0080] Example 14 Clinical evaluation of ceftiofur hydrochloride injection in the treatment of porcine contagious pleuropneumonia

[0081] 40 pigs with porcine contagious pleuropneumonia and 20 healthy pigs were divided into 3 groups, 20 in each group, and kept in isolation. The first group was the infected pig experimental group, and the dose was 5 mg·kg -1 The preparation B8 prepared in Example 10 was injected intramuscularly into the neck of pigs (body weight); Group 2 was the infected pig control group, at 5 mg·kg -1 The pigs were injected with normal saline in the neck muscle; the third group was the healthy pig control group, at 5 mg·kg -1 (body weight) Inject normal saline into the pig's neck muscle. Observe the pig's clinical manifestations for 7 consecutive days after a single administration, record the feed intake, body weight changes, body temperature changes, defecation, etc., and judge the drug efficacy according to the following method. If the pig dies, dissect and observe the pathological conditions of its organs.

[0082] Efficacy judgment:

[0083] Clinical cure: 3 days after administration, the body temperature, mental state, feeding, drinking, breathing and defecation of the test pigs returned to normal, and there was no recurrence after 7 days.

[0084] Clinical effectiveness: 3 days after administration, the test pigs' body temperature returned to normal, their mental state improved significantly, their feed intake increased, and they had a slight cough or mild abdominal breathing.

[0085] Clinical ineffectiveness: 3 days after administration, the test pigs' body temperature, mental state, feeding, drinking, breathing and defecation indicators did not improve and even worsened.

[0086] The therapeutic effect of ceftiofur hydrochloride injection on porcine contagious pleuropneumonia is shown in Table 8. -1 When the drug is administered according to body weight, it has an obvious therapeutic effect on porcine contagious pleuropneumonia, with an effective rate of 95% and a cure rate of 90%, indicating that the ceftiofur hydrochloride injection prepared by the present invention has a good therapeutic effect on porcine contagious pleuropneumonia.

[0087] Table 8 The therapeutic effect of ceftiofur hydrochloride injection on porcine contagious pleuropneumonia

[0088]

[0089] Example 15 Clinical evaluation of ceftiofur hydrochloride injection in the treatment of postpartum metritis in dairy cows

[0090] Thirty dairy cows with postpartum metritis were divided into two groups, 15 in each group. The first group was the experimental group, and the dose was 2.2 mg·kg -1 (body weight) The preparation B8 prepared in Example 10 was injected intramuscularly once a day for 3 consecutive days; the second group was the control group, at 0.5 mg·kg -1 (body weight) intramuscular injection of cloprostenol injection was given on the 1st and 14th days; colposcopy was used to examine and calculate the cure rate.

[0091] The results showed that the cure rate of the experimental group was 85.6%, and the cure rate of the control group was 78.3%. The preparation B8 prepared in Example 10 had a better therapeutic effect on postpartum metritis in dairy cows than cloprostenol injection, indicating that the ceftiofur hydrochloride injection of the present invention has a significant therapeutic effect on postpartum metritis in dairy cows.

[0092] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. Ceftiofur hydrochloride injection, characterized in that: The invention is composed of the following components by weight: 5 to 11 parts of ceftiofur hydrochloride, 1 to 2 parts of a suspending emulsifier, 0.01 to 0.04 parts of an antioxidant, and 45 to 66 parts of an oil solvent; the suspending emulsifier is a lipophilic sucrose fatty acid ester and / or aluminum stearate; the oil solvent is one or more of caprylic decanoic acid glyceride, refined coconut oil, and cottonseed oil.

2. The ceftiofur hydrochloride injection according to claim 1, characterized in that: The invention is composed of the following components by weight: 5-11 parts of ceftiofur hydrochloride, 1-2 parts of suspending emulsifier, 0.01-0.04 parts of antioxidant and 60-65 parts of oil solvent; the suspending emulsifier is lipophilic sucrose fatty acid ester and / or aluminum stearate; the oil solvent is caprylic decanoic acid glyceride and / or refined coconut oil.

3. The ceftiofur hydrochloride injection according to claim 2, characterized in that: The invention is composed of the following components by weight: 5-6 parts of ceftiofur hydrochloride, 2 parts of aluminum stearate, 0.02 parts of antioxidant BHA, and 64-65 parts of caprylic / capric glyceride.

4. The ceftiofur hydrochloride injection according to claim 2, characterized in that: The invention is composed of the following components by weight: 5-6 parts of ceftiofur hydrochloride, 1.5 parts of aluminum stearate, 0.5 parts of lipophilic sucrose fatty acid ester, 0.02 parts of antioxidant BHA, and 64-65 parts of caprylic decanoic acid glyceride.

5. The ceftiofur hydrochloride injection according to claim 2, characterized in that: The invention is composed of the following components by weight: 10-11 parts of ceftiofur hydrochloride, 1.5 parts of aluminum stearate, 0.04 parts of antioxidant BHA, and 61-62 parts of caprylic and capric glyceride.

6. The ceftiofur hydrochloride injection according to claim 2, characterized in that: The invention is composed of the following components by weight: 5-6 parts of ceftiofur hydrochloride, 2 parts of lipophilic sucrose fatty acid ester, 0.02 parts of antioxidant BHA, and 61-62 parts of caprylic and capric glyceride.

7. The ceftiofur hydrochloride injection according to claim 2, characterized in that: The invention is composed of the following components by weight: 5-6 parts of ceftiofur hydrochloride, 1.2 parts of aluminum stearate, 0.02 parts of antioxidant BHA, 28-29 parts of refined coconut oil, and 35-36 parts of caprylic and capric glyceride.

8. The method for preparing the ceftiofur hydrochloride injection according to any one of claims 1 to 7, characterized in that: The following steps are involved: Take ceftiofur hydrochloride, disperse it in part of the oil solvent to obtain a crude ceftiofur hydrochloride solution; take a suspending emulsifier, disperse it in the remaining oil solvent, heat it to 75-125° C., stop heating, wait for it to cool to 50-65° C., add an antioxidant, stir to completely dissolve, wait for it to cool to 35-45° C., add the crude ceftiofur hydrochloride solution, stir evenly, wait for it to cool to 25-35° C., transfer it into a high-pressure homogenizer, first circulate homogenize at a pressure of 5-20 MPa for 10-30 minutes, and then repeat the homogenization twice at a pressure of 10-30 MPa to obtain the ceftiofur hydrochloride injection.

9. The method for preparing ceftiofur hydrochloride injection according to claim 8, characterized in that: The following steps are involved: Take ceftiofur hydrochloride, disperse it in part of the oil solvent to obtain a crude ceftiofur hydrochloride solution; take a suspending emulsifier, disperse it in the remaining oil solvent, heat it to 75-120° C., stop heating, wait for it to cool to 55-60° C., add an antioxidant, stir to completely dissolve, wait for it to cool to 35° C., add the crude ceftiofur hydrochloride solution, stir for 15-20 minutes, wait for it to cool to 30° C., transfer it into a high-pressure homogenizer, first circulate homogenize at a pressure of 8-15 MPa for 15-30 minutes, and then repeat the homogenization twice at a pressure of 15-25 MPa to obtain ceftiofur hydrochloride injection.

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