A compound procaine penicillin suspension injection and a preparation method thereof

Abstract

The application discloses a compound procaine penicillin suspension injection, which contains the following raw and auxiliary materials in 100 mL of the suspension injection: 5-30 g of procaine penicillin, 5-30 g of dihydrostreptomycin sulfate, 0.5-1.5 g of non-ionic surfactant, and the balance of injection solvent; the non-ionic surfactant contains a diaryl secondary amine structure, a cyclohexane structure, an ester group and a hydroxyl group, has good compatibility with the procaine penicillin, the dihydrostreptomycin sulfate and the injection solvent, can not only reduce the interfacial tension of each component in the mixture system, increase the dispersibility and stability of the mixture system, but also improve the sedimentation volume ratio and the needle passability of the suspension injection; and the application further discloses a preparation method of the suspension injection. The compound procaine penicillin suspension injection provided by the application is safe and effective, has a high sedimentation volume ratio, excellent stability, dispersibility and needle passability.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of veterinary medicine, and particularly relates to a compound procaine penicillin suspension injection and a preparation method thereof. BACKGROUND

[0002] Procaine penicillin is a procaine salt of penicillin, and the antibacterial active ingredient is penicillin. Penicillin is a bactericidal antibiotic with strong antibacterial activity. Its antibacterial mechanism is mainly to inhibit the synthesis of bacterial cell wall mucin. The sensitive bacteria in the growth period are vigorous in division. The cell wall is in the biosynthesis period, and under the action of penicillin, the synthesis of mucin is blocked and the cell wall cannot be formed, resulting in cell membrane rupture and death under the action of osmotic pressure. Penicillin is a narrow-spectrum antibiotic, mainly acting on various gram-positive bacteria and a few gram-negative cocci. The main sensitive bacteria include Staphylococcus, Streptococcus, swine erysipelas bacillus, corynebacterium, tetanus clostridium, actinomycetes, anthrax bacillus, and spirochetes. Dihydrostreptomycin sulfate belongs to the aminoglycoside antibiotic, which can cause abnormal protein synthesis by interfering with the protein synthesis process of bacteria, hinder the release of synthesized protein, and also can enhance the permeability of bacterial cell membrane to cause the leakage of some important physiological substances, and finally cause bacterial death. Dihydrostreptomycin has antibacterial effect on tubercle bacillus and various gram-negative bacilli such as Escherichia coli, Salmonella, Brucella, Pasteurella, Shigella dysenteriae, and nose fistula bacillus.

[0003] As a commonly used dosage form, injection has fast effect, accurate dose, reliable action, and is not affected by gastrointestinal factors, and can produce positioning, targeting and long-acting effect, and is suitable for individuals who cannot take oral administration and drugs that cannot be taken orally.

[0004] The patent with the application number CN201110188995.5 provides a procaine penicillin-sulfate dihydrostreptomycin suspension injection, each 100 mL injection contains: procaine penicillin 15-25 g, sulfate dihydrostreptomycin 20-30 g, suspending agent polyvinylpyrrolidone 0.3-0.7 g, antioxidant formaldehyde sodium bisulfite 0.1-0.3 g, preservative benzyl alcohol 0.3-0.7 g. The invention has strong bactericidal effect by the synergistic bactericidal effect of procaine penicillin and sulfate dihydrostreptomycin, and can be used for treating livestock streptococcal infection and mixed infection of gram-positive bacteria and gram-negative bacteria. The clinical curative effect is good. At the same time, the invention has significant curative effect on gastrointestinal tract, respiratory tract and urinary tract infections, mastitis, uterine abscess, complex fracture, joint pain, leg injury, arthritis and prevention of postpartum infection caused by procaine penicillin and sulfate dihydrostreptomycin sensitive microorganism flora such as escherichia coli, erysipelas bacillus, clostridium, streptococcus, staphylococcus, salmonella and pasteurella. The patent with the application number CN201210070928.8 provides a veterinary procaine penicillin-sulfate dihydrostreptomycin suspension injection, the preparation composition of which comprises: procaine penicillin 5%-30%(w / v), sulfate dihydrostreptomycin 5%-30%(w / v), preservative 0.02%-0.2%(w / v), dispersing agent 0.5%-15%(w / v), suspending agent 0.2%-20%(w / v), stabilizer and local analgesic 0.5%-2%(w / v), antioxidant 0.02%-0.5%(w / v), and buffer is used to adjust the pH value of the system to 5.0-8.0. The invention improves the convenience of drug use and expands the antibacterial spectrum of the drug by forming a compound preparation of procaine penicillin and sulfate dihydrostreptomycin. At the same time, the invention greatly saves the cost and reduces the risk of damage to the animal body caused by the dispersion medium after injection by preparing the suspension injection with water as the dispersion medium. The above-mentioned patents mix procaine penicillin and sulfate dihydrostreptomycin to expand the antibacterial spectrum of the drug and facilitate the clinical use of the drug. However, the non-homogeneous liquid preparation formed by dispersing the poorly soluble or slightly soluble drug in the dispersion medium has low sedimentation volume ratio, poor stability, poor dispersibility and poor needle passability, and the product quality is poor, which is a problem that needs to be solved by those skilled in the art.

[0005] With the development of livestock breeding, intensive breeding of pigs, chickens, cattle, sheep and other animals is more common. Under such intensive feeding conditions, respiratory diseases, mixed infections and secondary mixed infections are more likely to occur and spread. Therefore, it is particularly important to reduce the disease probability of animals and promote the healthy growth of animals. Therefore, it is crucial to develop a compound procaine penicillin suspension injection with excellent stability, dispersibility and needle passability according to the condition of animals, combined with the characteristics of drug preparation and pharmacology, and with the help of new technologies of modern drug research. SUMMARY

[0006] In view of the deficiencies of the prior art, the present application aims to provide a compound procaine penicillin suspension injection, which is safe and effective, has a high sedimentation volume ratio, excellent stability, dispersibility and needle passability, and a preparation method of the suspension injection.

[0007] The technical scheme adopted by the present application to achieve the above-mentioned purpose is:

[0008] A compound procaine penicillin suspension injection, the suspension injection contains the following raw and auxiliary materials per 100 mL: procaine penicillin 5-30 g, dihydrostreptomycin sulfate 5-30 g, non-ionic surfactant 0.5-1.5 g, and the balance is injection vehicle;

[0009] The preparation method of the non-ionic surfactant is as follows:

[0010] S1, disperse 4,4'-iminobenzoic acid in N,N-dimethylformamide, add p-toluenesulfonic acid and 6-hydroxyhexanoic acid in sequence, heat to 65-85 DEG C, react for 3-7 h, cool to room temperature, filter, take the filtrate, extract, and reduce pressure distillation to obtain an intermediate;

[0011] S2, disperse the intermediate obtained in step S1 in N,N-dimethylformamide, add chromium acetate and 2-cyclohexyl oxirane in sequence, heat to 75-95 DEG C, stir for 6-12 h, cool to room temperature, filter, take the filtrate, extract, and reduce pressure distillation to obtain the non-ionic surfactant;

[0012] The synthesis route of the non-ionic surfactant is as follows:

[0013]

[0014] The present application uses 4,4'-iminobenzoic acid and 6-hydroxyhexanoic acid as raw materials, and esterification reaction occurs under the action of p-toluenesulfonic acid to obtain an intermediate; the intermediate structure contains a carboxyl group, and ring-opening reaction occurs with 2-cyclohexyl oxirane under the action of chromium acetate to obtain the non-ionic surfactant.

[0015] In order to obtain the nonionic surfactant and ensure product consistency, the molar ratio of 4, 4'-iminodibenzoic acid, p-toluenesulfonic acid and 6-hydroxyhexanoic acid in step S1 is 1:0.02-0.08:1.5-2.5, and the addition amount of 4, 4'-iminodibenzoic acid in the N, N-dimethylformamide is 0.2-0.4 g / mL; the molar ratio of the intermediate, chromium acetate and 2-cyclohexyl oxirane in step S2 is 1:0.015-0.035:1.8-2.1, and the addition amount of the intermediate in the N, N-dimethylformamide is 0.25-0.35 g / mL.

[0016] Further, the injection solvent is one or two or more of soybean oil for injection, ethyl oleate, glyceryl triacetate, and medium-chain triglyceride.

[0017] The application also provides a preparation method of the compound procaine penicillin suspension injection, which comprises the following steps: under stirring, part of the injection solvent is heated to 60-90 DEG C, and the nonionic surfactant is added, and after uniform stirring, the temperature is cooled to room temperature, and then the procaine penicillin and the dihydrostreptomycin sulfate are sequentially added, and after uniform stirring, the remaining injection solvent is added, and after mixing, grinding, filling, sterilization, the compound procaine penicillin suspension injection is obtained; the volume of the part of the injection solvent is 40-60% of the total volume of the injection solvent.

[0018] The application has the following beneficial effects: the nonionic surfactant is obtained through esterification reaction and epoxide ring-opening reaction in sequence, and the surfactant has excellent dispersibility, stability and emulsifying property; the nonionic surfactant contains a diaryl secondary amine structure, a cyclohexane structure, an ester group and a hydroxyl group, and has good compatibility with the procaine penicillin, the dihydrostreptomycin sulfate and the injection solvent, which can not only reduce the interfacial tension of each component in the mixture system, increase the dispersibility, high-temperature stability and light stability of the mixture system, but also improve the sedimentation volume ratio and the needle passability of the suspension injection; the compound procaine penicillin suspension injection provided by the application is safe and effective, has a high sedimentation volume ratio, excellent stability, dispersibility and needle passability. DETAILED DESCRIPTION

[0019] The technical solutions in the embodiments of the application will be clearly and completely described below in combination with the embodiments of the application. Obviously, the described embodiments are only part of the embodiments of the application, rather than all the embodiments of the application. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the application. The raw materials used in the following embodiments are all commercially available products.

[0020] Embodiment 1

[0021] A preparation method of a non-ionic surfactant, comprising the following steps:

[0022] S1, dispersing 4, 4'-iminodibenzoic acid in N, N-dimethylformamide, sequentially adding p-toluenesulfonic acid and 6-hydroxyhexanoic acid, heating to 70 DEG C, reacting for 5h, cooling to room temperature, filtering, taking the filtrate, adding deionized water and ethyl acetate for extraction, taking the organic phase, and distilling under reduced pressure to obtain an intermediate, wherein the molar ratio of 4, 4'-iminodibenzoic acid, p-toluenesulfonic acid and 6-hydroxyhexanoic acid is 1:0.04:1.8, the addition amount of 4, 4'-iminodibenzoic acid in N, N-dimethylformamide is 0.28g / mL, and the volume ratio of N, N-dimethylformamide, deionized water and ethyl acetate is 1:1:2;

[0023] S2, dispersing the intermediate obtained in step S1 in N, N-dimethylformamide, sequentially adding chromium acetate and 2-cyclohexyl oxirane, heating to 80 DEG C, stirring for 10h, cooling to room temperature, filtering, taking the filtrate, adding deionized water and ethyl acetate for extraction, taking the organic phase, and distilling under reduced pressure to obtain the non-ionic surfactant, wherein the molar ratio of the intermediate, chromium acetate and 2-cyclohexyl oxirane is 1:0.028:1.9, the addition amount of the intermediate in N, N-dimethylformamide is 0.33g / mL, and the volume ratio of N, N-dimethylformamide, deionized water and ethyl acetate is 1:1:2;

[0024] The synthesis route of the non-ionic surfactant is as follows:

[0025]

[0026] The nuclear magnetic resonance result of the non-ionic surfactant is as follows: 1 H NMR (300 MHz, CDCl3) δ 8.36 (s, 1H), 7.74 (d, 4H), 7.43 (d, 4H), 5.37 (s, 2H), 4.30-4.34 (m, 6H), 4.09 (d, 2H), 3.96-3.99 (m, 2H), 2.32 (t, 4H), 1.75-1.79 (m, 4H), 1.62-1.66 (m, 8H), 1.50-1.54 (m, 4H), 1.43-1.46 (m, 8H), 1.33-1.38 (m, 6H), 1.25-1.29 (m, 4H).

[0027] Example 2

[0028] A compound procaine penicillin suspension injection, 100mL of which contains the following raw and auxiliary materials: procaine penicillin 20g, dihydrostreptomycin sulfate 20g, non-ionic surfactant prepared in example 1 1g, and the rest is injection solvent, wherein the injection solvent is ethyl oleate.

[0029] A preparation method of compound procaine penicillin suspension injection, comprising the following steps:

[0030] Under stirring, part of ethyl oleate (the volume of part of ethyl oleate is 50% of the total volume of ethyl oleate) is heated to 80℃, and the non-ionic surfactant prepared in Example 1 is added. After stirring uniformly, it is cooled to room temperature. Then procaine penicillin and dihydrostreptomycin sulfate are added in sequence. After stirring uniformly, the remaining ethyl oleate is added. After mixing uniformly, it is ground for 10 min by an emulsifying machine. After sterilization by filling and sealing, it is obtained.

[0031] Example 3

[0032] A preparation method of compound procaine penicillin suspension injection is prepared according to the method of Example 2, with the difference that 0.5 g of non-ionic surfactant is contained in 100 mL of suspension injection.

[0033] Example 4

[0034] A preparation method of compound procaine penicillin suspension injection is prepared according to the method of Example 2, with the difference that 1.5 g of non-ionic surfactant is contained in 100 mL of suspension injection.

[0035] Comparative Example 1

[0036] A preparation method of compound procaine penicillin suspension injection is prepared according to the method of Example 2, with the difference that the non-ionic surfactant is replaced by a blend of lecithin, Tween-80 and vitamin E (0.5 g of lecithin, 0.5 g of Tween-80 and 0.5 g of vitamin E).

[0037] Comparative Example 2

[0038] A preparation method of compound procaine penicillin suspension injection is prepared according to the method of Example 2, with the difference that the non-ionic surfactant is replaced by Tween-80.

[0039] The compound procaine penicillin suspension injection prepared in Examples 2-4 is subjected to acute toxicity test. 90 healthy mice (KM grade, purchased from Henan Skbets Biotechnology Co., Ltd.) are taken, and the experimental mice (weight (20.2±0.3) g) are randomly divided into three groups, each group of 30, and each group is further divided into 6 groups, and each group is injected intramuscularly at 0.05 mL, 0.1 mL, 0.2 mL, 0.4 mL, 0.6 mL and 1 mL per kilogram of body weight, respectively, and observed for 3 days after administration. The first group is injected with the compound procaine penicillin suspension injection prepared in Example 2, the second group is injected with the compound procaine penicillin suspension injection prepared in Example 3, and the third group is injected with the compound procaine penicillin suspension injection prepared in Example 4. After 3 days of observation, none of the mice died, and no adverse reactions were found. The results show that the compound procaine penicillin suspension injection provided by the application is safe at a dosage of 0.05 mL to 1 mL per kilogram of body weight.

[0040] The compound procaine penicillin suspension injection prepared in Examples 2-4 and Comparative Examples 1-2 was subjected to relevant performance detection according to the requirements of the Chinese Veterinary Pharmacopoeia for suspensions. For the sedimentation volume ratio detection, a stoppered cylinder was used to measure 50 mL of the sample, tightly stoppered, and shaken vigorously for 1 min. The initial height H0 of the suspension was recorded, and the sample was left to stand for 5 h. The final height H of the suspension was recorded, and the sedimentation volume ratio was calculated according to the following formula: sedimentation volume ratio = H / H0. For the needle passability detection, the sample was shaken and then 5 mL of air was injected into the sample using a 7-gauge needle. The volume of the suspension sucked in was measured within 1 min. For the dispersibility detection, the sample was centrifuged at a speed of 4000 r / min for 15 min, shaken, and then the dispersion state was observed. For the high-temperature stability detection, the sample was placed at (60±2) °C for 3 months. Samples were taken at 0 months and 3 months, respectively. High-performance liquid chromatography was used for analysis, and the relative content of the active ingredients was calculated and the appearance of the suspension was observed. For the light stability test, the sample was placed in a stoppered cylinder and placed in a light source with an illumination of 4500±500 Lx. Samples were taken at 0 days and 15 days, respectively. High-performance liquid chromatography was used for analysis, and the relative content of the active ingredients was calculated and the appearance of the suspension was observed. For the accelerated test, the sample was placed in a stoppered cylinder and placed in an electric heating constant temperature incubator at a temperature of (30±2) °C and a relative humidity of (55±5) % for long-term storage for 12 months. Samples were taken at 0 months, 6 months, and 12 months, respectively. High-performance liquid chromatography was used for analysis, and the relative content of the active ingredients was calculated and the appearance of the suspension was observed. The experimental results are shown in Table 1. As can be seen from the table, the sedimentation volume ratio, needle passability, dispersibility, and stability of the compound procaine penicillin suspension injection prepared in Examples 2-4 are significantly better than those of Comparative Examples 1-2. The compound procaine penicillin suspension injection prepared in Examples 2-4 was subjected to high-temperature stability, light stability, and accelerated test. At the end of the test, the appearance of the sample, the relative content of procaine penicillin, and the relative content of dihydrostreptomycin sulfate did not change significantly, showing excellent stability. Compared with the simple mixing of multi-component surfactants, the addition of a single-component non-ionic surfactant (prepared in Example 1) to the raw material components can significantly improve the comprehensive performance of the compound procaine penicillin suspension injection.

[0041] Table 1: Results of relevant performance detection

[0042]

[0043]

[0044] The compound procaine penicillin suspension injection prepared in Examples 2-4 and Comparative Examples 1-2 was subjected to clinical efficacy test. 60 cows with natural mastitis (from a breeding farm in Kaifeng, Henan Province) were randomly divided into 3 groups, 20 cows in each group. The cows were deprived of food 16 hours before the test and only allowed to drink water. The cows were injected with the drug at a dose of 0.2 mL per kg of body weight by intramuscular injection once a day, and the treatment was continued for 5 days. The efficacy was determined according to the clinical symptoms, and the results were classified into 3 categories: cured: the redness, heat and pain of the affected area of the udder disappeared, the clots and flocculent matter in the milk disappeared, the milk color and milk yield returned to normal, the spirit and appetite were normal, and no recurrence was observed after 7 days of drug withdrawal; effective: the clinical symptoms disappeared, but recurrence was observed after drug withdrawal; ineffective: the symptoms of the sick cows did not disappear, and the disease even worsened. The results are shown in Table 2. As can be seen from the table, the effective rate of the compound procaine penicillin suspension injection prepared in Examples 2-4 for treating mastitis in cows was 100%, and the cure rate was 95% or higher. The compound procaine penicillin suspension injection prepared in Example 2 had the best treatment effect, and the cure rate was 100%.

[0045] Table 2: Results of clinical efficacy test

[0046] Test item Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Efficiency 100％ 100％ 100％ 90％ 75％ Cure rate 100％ 95％ 95％ 80％ 60％ Inefficiency 0％ 0％ 0％ 10％ 25％

[0047] The present application uses 4,4'-iminodibenzoic acid and 6-hydroxyhexanoic acid as raw materials, and under the action of p-toluenesulfonic acid, an esterification reaction occurs to obtain an intermediate. The intermediate structure contains a carboxyl group, which undergoes ring-opening reaction with 2-cyclohexyl oxirane under the action of chromium acetate to obtain a non-ionic surfactant. The surfactant has excellent dispersibility, stability and emulsifying property, and has good compatibility with procaine penicillin, dihydrostreptomycin sulfate and injection solvent. It not only can reduce the interfacial tension of each component in the mixture system, increase the dispersibility, high-temperature stability and light stability of the mixture system, but also can improve the sedimentation volume ratio and needle passability of the suspension injection. The compound procaine penicillin suspension injection provided by the present application is safe and effective, has a high sedimentation volume ratio, excellent stability, dispersibility and needle passability.

[0048] Although the embodiments of the present application have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and alterations can be made thereto without departing from the principles and spirit of the present application, and the scope of the present application is defined by the appended claims and their equivalents.

Claims

1. A compound procaine penicillin suspension injection solution, characterized by, The suspension injection liquid contains the following raw and auxiliary materials per 100 mL: procaine penicillin 5-30 g, dihydrostreptomycin sulfate 5-30 g, non-ionic surfactant 0.5-1.5 g, and the rest is injection solvent; The structural formula of the non-ionic surfactant is as follows:

2. The compound procaine penicillin suspension injection of claim 1, characterized in that, The preparation method of the non-ionic surfactant is as follows: S1, disperse 4,4'-iminobenzoic acid in N,N-dimethylformamide, add p-toluenesulfonic acid and 6-hydroxyhexanoic acid in sequence, heat to 65-85℃, react for 3-7 h, cool to room temperature, filter, take the filtrate, extract, and distill under reduced pressure to obtain an intermediate; S2, disperse the intermediate obtained in step S1 in N,N-dimethylformamide, add chromium acetate and 2-cyclohexyl oxirane in sequence, heat to 75-95℃, stir for 6-12 h, cool to room temperature, filter, take the filtrate, extract, and distill under reduced pressure to obtain the non-ionic surfactant.

3. The compound procaine penicillin suspension injection of claim 2, characterized in that, The molar ratio of 4,4'-iminobenzoic acid, p-toluenesulfonic acid and 6-hydroxyhexanoic acid in step S1 is 1:0.02-0.08:1.5-2.

5.

4. The compound procaine penicillin suspension injection of claim 3, characterized in that, The addition amount of 4,4'-iminobenzoic acid in the N,N-dimethylformamide is 0.2-0.4 g / mL.

5. The compound procaine penicillin suspension injection of claim 2, characterized in that, The molar ratio of the intermediate, chromium acetate and 2-cyclohexyl oxirane in step S2 is 1:0.015-0.035:1.8-2.

1.

6. The compound procaine penicillin suspension injection of claim 5, characterized in that, The addition amount of the intermediate in the N,N-dimethylformamide is 0.25-0.35 g / mL.

7. The compound procaine penicillin suspension injection of claim 1, characterized in that, The injection solvent is one or more of soybean oil for injection, ethyl oleate, glyceryl triacetate, and medium-chain triglyceride.

8. The method for preparing a compound procaine penicillin suspension injection according to any one of claims 1-7, characterized in that, The method comprises the following steps: Under stirring, part of the injection solvent is heated to 60-90℃, the non-ionic surfactant is added, stirred uniformly, and then cooled to room temperature, then the procaine penicillin and dihydrostreptomycin sulfate are added in sequence, stirred uniformly, and then the rest of the injection solvent is added, mixed uniformly, ground, sealed, sterilized, and then obtained.

9. The method for preparing a compound procaine penicillin suspension injection according to claim 8, characterized in that, The volume of the part of the injection solvent is 40-60% of the total volume of the injection solvent.

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