Medicinal preparation for treating endometritis as well as preparation method and application thereof
By using drug preparations containing enzyme preparations, polysorbate 80 and N-acetylcysteine, the problem of low efficacy of recombinant staphylococase in the treatment of endometriitis is solved, achieving higher therapeutic effects and lower risk of drug resistance.
Patent Information
- Application Number
- CN202510415828.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-03
- Publication Date
- 2025-06-13
AI Technical Summary
In the prior art, recombinant lysed staphylococase is less effective in the first course of treatment for endometriitis, and antibiotic use may lead to problems with bacterial resistance and drug residues.
A pharmaceutical preparation is employed which includes an enzyme preparation, a first composition of polysorbate 80 and N-acetylcysteine and a second composition of recombinant lysostatic staphylococase, lysozyme, N-acetylcysteine and a water-soluble matrix. This preparation reduces the viscoelasticity of mucus and pus through perfusion fluid, and improves the efficiency of drug action on the infection foci through suppositories.
It significantly improves the first course of treatment for endometriitis, improves the cure rate and efficiency, and reduces bacterial resistance and drug residue risks.
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Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of veterinary drug preparation, and particularly relates to a pharmaceutical preparation for treating endometritis, a preparation method thereof, and an application thereof. Background Art
[0002] Endometritis in female cows is a reproductive disorder disease caused by damaged uterine mucosa. The causes of endometritis in female cows are as follows: First, it is caused by the infection of pathogenic microorganisms such as Escherichia coli, Salmonella, Staphylococcus aureus, Streptococcus, Corynebacterium pyogenes, and Bacillus licheniformis due to the lack of strict disinfection during operations such as vaginal examination, artificial insemination, parturition, and dystocia assistance. Second, dairy buffaloes suffering from birth canal injury, dystocia, abortion, uterine prolapse, vaginal prolapse, vaginitis, cervicitis, tuberculosis, brucellosis, etc. will also be complicated with endometritis. When dairy buffaloes suffer from endometritis, factors such as the production of toxins by pathogenic bacteria and damaged endometrium affect ovulation, egg viability, sperm survival, or embryo survival, resulting in repeated infertility, increasing breeding costs, affecting reproductive plans, and causing economic losses.
[0003] In the prior art, the intrauterine perfusion method of soluble powder of spectinomycin hydrochloride and lincomycin hydrochloride is used. If antibiotic drugs are used for a long time, not only will bacteria develop drug resistance, affecting the efficacy of the drugs, but also the drugs will remain in the body of female buffaloes, affecting the quality and safety of dairy products. When recombinant lysostaphin is used to replace antibiotics for uterine perfusion, it is found that recombinant lysostaphin has a good therapeutic effect on specific types of bacteria such as staphylococci, but has relatively poor effects on other types of bacteria. Further, the overall efficacy of the first course of treatment is poor, probably because the uterine body of dairy cows with endometritis contains more mucus and pus, and the uterine body structure hinders the drugs from acting on the infection site, resulting in a generally low overall efficacy of the first course of treatment. Summary of the Invention
[0004] The main object of the present invention is to provide a pharmaceutical preparation for treating endometritis, aiming to solve the defect that the overall efficacy of the first course of treatment of recombinant lysostaphin perfusion is generally low.
[0005] To achieve the above object, the present invention provides a pharmaceutical preparation for treating endometritis, and the pharmaceutical preparation includes a first composition and a second composition;
[0006] The raw materials of the first composition include: an enzyme preparation, polysorbate 80, and N-acetylcysteine;
[0007] The raw materials of the second composition include: recombinant lysostaphin, lysozyme, N-acetylcysteine, and a water-soluble matrix.
[0008] Preferably, in the above technical solution, the raw materials of the first composition are 15 - 38 g of enzyme preparation, 0.1 - 1 mL of polysorbate 80, and 1 - 5 g of N-acetylcysteine by mass; the raw materials of the second composition are 10 - 20% of recombinant lysostaphin, 15 - 25% of lysozyme, 1 - 10% of N-acetylcysteine, and 45 - 74% of water-soluble matrix by mass percentage.
[0009] According to the above technical solution,
[0010] Preferably, in the above technical solution, the raw materials of the first composition further include leonurine.
[0011] Preferably, in the above technical solution, by mass, the raw materials of the first composition are 15 - 38 g of enzyme preparation, 0.1 - 1 mL of polysorbate 80, 1 - 5 g of N-acetylcysteine, and 5 - 10 mg of leonurine.
[0012] Preferably, in the above technical solution, the enzyme preparation includes one or two of Bacillus subtilis fibrinolytic enzyme and bromelain.
[0013] According to the above technical solution, the principle of the perfusion fluid in this application to reduce the viscoelasticity of mucus and pus is as follows: (1) N-acetylcysteine destroys the mucin structure and degrades the pus protein complex, and polysorbate 80 further blocks the polymerization of mucus and pus, reducing the cohesion of molecules in the mucus, jointly achieving the reduction of the viscoelasticity of mucus and pus and creating conditions for its discharge; (2) Bacillus subtilis fibrinolytic enzyme or / and bromelain degrades the proteins in mucus and pus, synergistically reducing the viscoelasticity of mucus and pus with N-acetylcysteine and polysorbate 80. At the same time, Bacillus subtilis fibrinolytic enzyme or / and bromelain inhibits the secretion of inflammatory mediators, further reducing the production of pus from the source; (3) Leonurine enhances uterine contraction and softens the cervix at the same time, promoting the discharge of mucus and pus.
[0014] Preferably, in the above technical solution, the raw materials of the second composition further include: polysorbate 80, vitamin B3, polyhexamethylene biguanide, and trehalose.
[0015] Preferably, in the above technical solution, by mass percentage, the raw materials of the second composition are 13 - 15% of recombinant lysostaphin, 18 - 24% of lysozyme, 1 - 3% of polysorbate 80, 1 - 5% of vitamin B3, 1 - 6% of polyhexamethylene biguanide, 0.5 - 2% of trehalose, 1 - 3% of N-acetylcysteine, and 35 - 60% of glycerogelatin.
[0016] According to the above technical solution, the suppository of the present application has the following principles for improving the first course of endometritis: (1) First, uterine perfusion is performed using a perfusion solution to reduce the secretion of inflammatory mediators, promote the discharge of mucus and pus, and enhance the drug effect of the suppository on the infection site to improve the drug cure efficiency; (2) The polysorbate 80 component in the suppository promotes the phospholipid emulsification of the bacterial biofilm, disintegrates the biofilm matrix, and assists drug delivery with N-acetylcysteine, further promoting the action of recombinant staphylokinase and lysozyme on the infection site; (3) Recombinant staphylokinase and lysozyme can cut the pentaglycine peptide bond bridge structure in the peptidoglycan of the staphylococcal cell wall and cleave the β-1,4 bond between N-acetylmuramic acid and N-acetylglucosamine in the peptidoglycan molecule of the bacteria, causing the bacteria to lyse and die; (4) Polyhexamethylene biguanide and vitamin B3 inhibit the release of inflammatory factors, promote the regeneration of squamous epithelial cells, accelerate the healing speed of the inflammatory wound, and improve the cure efficiency.
[0017] A preparation method of a pharmaceutical preparation for treating endometritis, the preparation method comprising:
[0018] S1, preparing a perfusion solution with a first composition:
[0019] S11, dissolving leonurine in water to obtain a leonurine solution; dissolving Bacillus subtilis fibrinolytic enzyme in water to obtain a Bacillus subtilis fibrinolytic enzyme solution;
[0020] S12, adding NAC to water, heating to 25 - 30 °C for dissolution and adjusting the pH to 4.0 - 5.0, then adding polysorbate 80, stirring and mixing evenly, and slowly mixing the leonurine solution and the Bacillus subtilis fibrinolytic enzyme solution into the solution, making up the volume, and sterilizing to obtain the perfusion solution;
[0021] S2, preparing a suppository with a second composition:
[0022] S21, dissolving trehalose in water, and then adding recombinant staphylokinase and lysozyme to obtain a pretreatment solution;
[0023] S22, heating a water-soluble matrix to 38 - 40 °C, adding polysorbate 80, vitamin B3, and polyhexamethylene biguanide, stirring until completely dissolved, cooling to 25 - 30 °C, then adding the pretreatment solution and N-acetylcysteine and slowly stirring and dissolving, injecting into a suppository mold, and cooling and solidifying at 4 °C.
[0024] Preferably, in the above technical solution, the sterilization is to filter the solution after volume determination with a 0.22 μm filter membrane.
[0025] Preferably, in the above technical solution, the preparation of the water-soluble matrix includes: taking gelatin, shearing it, soaking it to fully swell, dissolving it in a water bath at 60-70 °C, adding glycerol and water and stirring until uniform, and controlling the water content to be 10-20% to obtain the water-soluble matrix glycerogelatin.
[0026] An application of a pharmaceutical preparation for treating endometritis as described above, the pharmaceutical preparation is an external medicine, and the usage method is to first use the perfusion solution prepared from the first composition, and then use the suppository prepared from the second composition.
[0027] Compared with the prior art, the present application has the following beneficial effects:
[0028] The pharmaceutical preparation of the present application can improve the treatment effect of endometritis in female buffaloes in the first treatment course. The principle is as follows: First, the enzyme preparation in the perfusion solution synergistically acts with N-acetylcysteine and polysorbate 80 to reduce the viscoelasticity of mucus and pus, and promote the discharge of mucus and pus; Second, after the perfusion treatment with the perfusion solution, the suppository is applied to promote the penetration of the suppository to the infection site. Recombinant lysostaphin can cut the pentaglycine peptide bond bridge structure in the peptidoglycan of the staphylococcal cell wall and combine with the β-1,4 bond between N-acetylmuramic acid and N-acetylglucosamine in the peptidoglycan molecule lysed by lysozyme to inhibit or kill Staphylococcus aureus, Escherichia coli, Salmonella, Streptococcus and Bacillus licheniformis. Detailed implementation mode
[0029] The technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts belong to the scope of protection of the present invention.
[0030] Example 1
[0031] A pharmaceutical preparation for treating endometritis, the pharmaceutical preparation includes a first composition and a second composition, and the raw materials of the first composition: 28 g of Bacillus subtilis fibrinolytic enzyme, 1 mL of polysorbate 80, 5 g of N-acetylcysteine, 10 mg of leonurine;
[0032] The raw materials of the second composition: 15% recombinant lysostaphin, 24% lysozyme, 5% vitamin B3, 3% trehalose, 6% polyhexamethylene biguanide, 3% polysorbate 80, 3% N-acetylcysteine, 41% glycerogelatin.
[0033] A preparation method of a pharmaceutical preparation for treating endometritis, the preparation method includes:
[0034] S1, preparing a perfusion solution:
[0035] S11. Dissolve 10 mg of leonurine in 5 mL of water to obtain a leonurine solution; dissolve 28 g of Bacillus subtilis fibrinolytic enzyme in 40 mL of water to obtain a Bacillus subtilis fibrinolytic enzyme solution.
[0036] S12. Take 5 g of N-acetylcysteine and add it to 40 mL of water. Place it in a 30°C water bath to dissolve and adjust the pH to 4.0 - 5.0. After complete dissolution, add 1 mL of polysorbate 80 and stir well. Pour the above solution into a 100 mL volumetric flask, and slowly add the leonurine solution and the Bacillus subtilis fibrinolytic enzyme solution to the volumetric flask. Add water to make up to 100 mL, and filter and sterilize with a 0.22 μm filter membrane to obtain the perfusion solution.
[0037] S2. Prepare the second composition:
[0038] S21. Dissolve 3% trehalose in water, and then add 15% recombinant lysostaphin and 24% lysozyme to obtain a pretreatment solution.
[0039] S22. Heat 41% of the water-soluble matrix to 40°C, add 3% of polysorbate 80, 5% of vitamin B3, and 6% of polyhexamethylene biguanide, and stir until completely dissolved. After cooling to 30°C, add the pretreatment solution and 3% of N-acetylcysteine and stir slowly to dissolve. Inject it into a suppository mold and cool and solidify at 4°C. The preparation method of the glycerogelatin: Take 20 g of gelatin, cut it, soak it to fully swell, dissolve it in a 70°C water bath, then add 80 g of glycerol and an appropriate amount of water and stir until uniform, controlling the water content to be 10 - 20% to obtain the water-soluble matrix glycerogelatin.
[0040] The application of a pharmaceutical preparation for treating endometritis. The pharmaceutical preparation is an external medicine. The usage method is to first perform uterine perfusion on the buffalo with endometritis using a uterine infusion device and the perfusion solution. 4 hours after the perfusion solution is administered, insert the suppository into the uterus, administer the medicine once every two days, and use it continuously three times as a course of treatment.
[0041] Example 2
[0042] A pharmaceutical preparation for treating endometritis, the pharmaceutical preparation includes a first composition and a second composition. The raw materials of the first composition: 20 g of Bacillus subtilis fibrinolytic enzyme, 0.5 mL of polysorbate 80, 3 g of N-acetylcysteine, 6 mg of leonurine;
[0043] The raw materials of the second composition: 13% recombinant lysostaphin, 20% lysozyme, 3% vitamin B3, 1% trehalose, 5% polyhexamethylene biguanide, 1% polysorbate 80, 1% N-acetylcysteine, 56% glycerogelatin.
[0044] Example 3
[0045] A pharmaceutical preparation for treating endometritis, the pharmaceutical preparation comprising a first composition and a second composition, the raw materials of the first composition: 28 g of bromelain, 1 mL of polysorbate 80, 5 g of N-acetylcysteine, 10 mg of leonurine;
[0046] The raw materials of the second composition: 15% recombinant lysostaphin, 24% lysozyme, 5% vitamin B3, 3% trehalose, 6% polyhexamethylene biguanide, 3% polysorbate 80, 3% N-acetylcysteine, 41% glycerogelatin.
[0047] A preparation method of a pharmaceutical preparation for treating endometritis, the preparation method comprising:
[0048] S1, preparing an infusion solution:
[0049] S11, dissolving 10 mg of leonurine in 5 mL of water to obtain a leonurine solution; dissolving 28 g of bromelain in 40 mL of water to obtain a bromelain solution;
[0050] S12, adding 5 g of N-acetylcysteine to 40 mL of water, placing it in a water bath at 30 °C to dissolve and adjusting the pH to 4.0 - 5.0; after complete dissolution, adding 1 mL of polysorbate 80 and stirring evenly; pouring the above solution into a 100 mL volumetric flask, and slowly adding the leonurine solution and the bromelain solution to the volumetric flask, adding water to make up the volume to 100 mL, and filtering and sterilizing with a 0.22 μm filter membrane to obtain the infusion solution.
[0051] S2, preparing the second composition:
[0052] S21, dissolving 3% of trehalose in water, and then adding 15% of recombinant lysostaphin and 24% of lysozyme to obtain a pretreatment solution;
[0053] S22, heating 41% of a water-soluble matrix to 40 °C, adding 3% of polysorbate 80, 5% of vitamin B3, and 6% of polyhexamethylene biguanide, stirring until completely dissolved, cooling to 30 °C, then adding the pretreatment solution and 3% of N-acetylcysteine and slowly stirring and dissolving, injecting into a suppository mold, and cooling and solidifying at 4 °C; the preparation method of the glycerogelatin: taking 20 g of gelatin, shearing and soaking it to fully swell, dissolving it in a water bath at 70 °C, then adding 80 g of glycerol and an appropriate amount of water and stirring until uniform, controlling the water content to be 10 - 20% to obtain the water-soluble matrix glycerogelatin.
[0054] Example 4
[0055] A pharmaceutical preparation for treating endometritis, the pharmaceutical preparation comprising a first composition and a second composition, the raw materials of the first composition: 20 g of bromelain, 0.5 mL of polysorbate 80, 3 g of N-acetylcysteine, 6 mg of leonurine;
[0056] Raw materials of the second composition: 13% recombinant lysostaphin, 20% lysozyme, 3% vitamin B3, 1% trehalose, 5% polyhexamethylene biguanide, 1% polysorbate 80, 1% N-acetylcysteine, 56% glycerogelatin.
[0057] Example 5
[0058] A pharmaceutical preparation for treating endometritis, the pharmaceutical preparation comprising a first composition and a second composition, and the raw materials of the first composition: 15 g of bromelain, 13 g of Bacillus subtilis fibrinolytic enzyme, 1 mL of polysorbate 80, 5 g of N-acetylcysteine, 10 mg of leonurine;
[0059] Raw materials of the second composition: 15% recombinant lysostaphin, 23% lysozyme, 3% vitamin B3, 3% trehalose, 5% polyhexamethylene biguanide, 1% polysorbate 80, 1% N-acetylcysteine, 49% glycerogelatin.
[0060] A preparation method of a pharmaceutical preparation for treating endometritis, the preparation method comprising:
[0061] S1, preparing a perfusion solution:
[0062] S11, dissolving 10 mg of leonurine in 5 mL of water to obtain a leonurine solution; dissolving 13 g of Bacillus subtilis fibrinolytic enzyme in 25 mL of water to obtain a Bacillus subtilis fibrinolytic enzyme solution; dissolving 15 g of bromelain in 25 mL of water to obtain a bromelain solution;
[0063] S12, adding 5 g of N-acetylcysteine to 40 mL of water, placing it in a water bath at 30 °C to dissolve and adjusting the pH to 4.0 - 5.0; after complete dissolution, adding 1 mL of polysorbate 80 and stirring evenly; pouring the above solution into a 100 mL volumetric flask, and slowly adding the leonurine solution, Bacillus subtilis fibrinolytic enzyme solution and bromelain solution into the volumetric flask, adding water to a constant volume of 100 mL, and filtering and sterilizing with a 0.22 μm filter membrane to obtain a perfusion solution.
[0064] S2, preparing the second composition:
[0065] S21, dissolving 3% of trehalose in water, and then adding 15% of recombinant lysostaphin and 23% of lysozyme to obtain a pretreatment solution;
[0066] S22. Heat 49% water-soluble matrix to 40°C, add 1% polysorbate 80, 3% vitamin B3, and 5% polyhexamethylene biguanide, stir until completely dissolved, cool to 30°C, then add the pretreatment solution and 3% N-acetylcysteine and stir slowly to dissolve. Inject into a suppository mold and cool and solidify at 4°C; Preparation method of the glycerogelatin: Take 20 g of gelatin, cut and soak it to fully swell, dissolve it in a water bath at 70°C, then add 80 g of glycerol and an appropriate amount of water and stir until uniform, controlling the water content to be 10 - 20% to obtain the water-soluble matrix glycerogelatin.
[0067] Comparative Example 1
[0068] A pharmaceutical preparation for treating endometritis, the raw materials of the pharmaceutical preparation are as follows: 30 g of 800U recombinant lysostaphin freeze-dried powder and 100 mL of water are mixed.
[0069] Comparative Example 2
[0070] A pharmaceutical preparation for treating endometritis, the raw materials of the pharmaceutical preparation are as follows: 30 g of lysozyme and 100 mL of water.
[0071] Comparative Example 3
[0072] A pharmaceutical preparation for treating endometritis, the raw materials of the pharmaceutical preparation are as follows:
[0073] 15% recombinant lysostaphin, 24% lysozyme, 3% trehalose, and 58% glycerogelatin.
[0074] Comparative Example 4
[0075] A pharmaceutical preparation for treating endometritis, the raw materials of the pharmaceutical preparation are as follows:
[0076] 15% recombinant lysostaphin, 24% lysozyme, 3% trehalose, 3% polysorbate 80, 3% N-acetylcysteine, and 52% glycerogelatin.
[0077] A preparation method of a pharmaceutical preparation for treating endometritis, the preparation method includes:
[0078] S1. Prepare suppositories:
[0079] S11. Dissolve 3% trehalose in water, then add 15% recombinant lysostaphin and 24% lysozyme to obtain a pretreatment solution;
[0080] S12. Heat 52% water-soluble matrix to 40°C, add 3% polysorbate 80, stir until completely dissolved, cool to 30°C, then add the pretreatment solution and 3% N-acetylcysteine, stir slowly to dissolve, inject into suppository molds, and cool and solidify at 4°C; The preparation method of the glycerogelatin: Take 20 g of gelatin, cut and soak it to fully swell, dissolve it in a water bath at 70°C, then add 80 g of glycerol and an appropriate amount of water, stir until uniform, and control the water content to be 10-20% to obtain the water-soluble matrix glycerogelatin.
[0081] Control Example 5
[0082] A pharmaceutical preparation for treating endometritis, the pharmaceutical preparation includes a first composition and a second composition, the raw materials of the first composition: 1 mL of polysorbate 80, 5 g of N-acetylcysteine and 10 mg of leonurine.
[0083] The raw materials of the second composition: 15% recombinant lysostaphin, 24% lysozyme, 3% trehalose, 3% polysorbate 80, 3% N-acetylcysteine and 52% glycerogelatin.
[0084] A preparation method of a pharmaceutical preparation for treating endometritis, the preparation method includes:
[0085] S1. Prepare the perfusion solution:
[0086] S11. Dissolve 10 mg of leonurine in 5 mL of water to obtain a leonurine solution.
[0087] S12. Add 5 g of N-acetylcysteine to 40 mL of water, dissolve it in a water bath at 30°C and adjust the pH to 4.0 - 5.0; after complete dissolution, add 1 mL of polysorbate 80 and stir well; pour the above solution into a 100 mL volumetric flask, and slowly add the leonurine solution and the Bacillus subtilis fibrinolytic enzyme solution into the volumetric flask, add water to make up to 100 mL, and filter and sterilize with a 0.22 μm filter membrane to obtain the perfusion solution.
[0088] S2. Prepare the second composition:
[0089] S21. Dissolve 3% trehalose in water, then add 15% recombinant lysostaphin and 24% lysozyme to obtain the pretreatment solution.
[0090] S22. Heat 52% water-soluble matrix to 40°C, add 3% polysorbate 80, stir until completely dissolved, cool to 30°C, then add the pretreatment solution and 3% N-acetylcysteine and stir slowly to dissolve. Inject into a suppository mold and cool and solidify at 4°C. Preparation method of the glycerogelatin: Take 20 g of gelatin, cut and soak it until fully swollen, dissolve it in a water bath at 70°C, then add 80 g of glycerol and an appropriate amount of water and stir until uniform, controlling the water content to be 10 - 20% to obtain the water-soluble matrix glycerogelatin.
[0091] Experimental subjects:
[0092] During the period from January 2023 to June 2024, 216 dairy cows suffering from endometritis in three buffalo farms in Guangxi Zhuang Autonomous Region. Characteristics of dairy cows with endometritis: When lying down or in heat, there is a lot of turbid mucus with flocs discharged from the vagina; during vaginal examination, the external orifice of the cervix is relaxed, the mucosa is congested, swollen and diffusely inflamed, or there are ulcers, or the cervical mucosa is swollen and there is tissue hyperplasia.
[0093] Treat the above-mentioned dairy cows with endometritis using the pharmaceutical preparations of Example 1, Example 3 and Example 5 and the pharmaceutical preparations of Comparative Example 1 - 5 respectively.
[0094] Medication methods for dairy cows with endometritis in Example 1, Example 3, Example 5 and Comparative Example 5: Before treatment, disinfect the insemination device with alcohol and ultraviolet light. Drive the diseased female cow to a fixed rack for fixation and fix the cow's tail. After cleaning the cow dung at the anus, rinse the vulva of the cow with benzalkonium bromide disinfectant and dry it. First, use the insemination device to perfuse the uterine cavity with the perfusion solution. After 4 hours of perfusion, then wear long-arm gloves and insert the suppository deep into the uterus. Repeat every 2 days, and administer the medicine 3 times in the first course of treatment.
[0095] Medication methods for dairy cows with endometritis in Comparative Example 1 and Comparative Example 2: Before treatment, disinfect the insemination device with alcohol and ultraviolet light. Drive the diseased female cow to a fixed rack for fixation and fix the cow's tail. After cleaning the cow dung at the anus, rinse the vulva of the cow with benzalkonium bromide disinfectant and dry it. Use the insemination device to perfuse the uterine cavity with the pharmaceutical preparation. Repeat every 2 days, and administer the medicine 3 times in the first course of treatment.
[0096] Medication methods for dairy cows with endometritis in Comparative Example 3 and Comparative Example 4: Before treatment, disinfect the insemination device with alcohol and ultraviolet light. Drive the diseased female cow to a fixed rack for fixation and fix the cow's tail. After cleaning the cow dung at the anus, rinse the vulva of the cow with benzalkonium bromide disinfectant and dry it. Wear long-arm gloves and insert the suppository deep into the uterus. Repeat every 2 days, and administer the medicine 3 times in the first course of treatment.
[0097] The judgment criteria are as follows:
[0098] (1) Cure: All symptoms related to endometritis completely disappear, clear and transparent secretions flow out of the vagina, and the uterine function returns to normal without inflammatory reaction;
[0099] (2) Effective: Symptoms related to endometritis basically disappear, there is a slight inflammation in the secretions flowing out of the vagina, the uterine function basically returns to normal, the inflammatory reaction is reduced, and the cervix becomes soft and smaller;
[0100] (3) Ineffective: There is no obvious improvement in both the symptoms related to endometritis and the uterine function, or the vaginal discharge is purulent, the uterine inflammation shows no change or worsens, showing no change compared with before medication, or even deteriorates.
[0101] The treatment effectiveness and effective rate of each group of dairy cows with endometritis after medication are shown in Table 1.
[0102] Table 1 Treatment effectiveness of each group of dairy cows with endometritis
[0103]
[0104]
[0105] As can be seen from Table 1, by using the perfusion fluid and suppository of the method of the present invention, after one treatment course, the treatment effective rate of dairy cows with endometritis can rise above 88%. Compared with the effective rate of 44.44% in Comparative Example 1 where only recombinant lysostaphin was used for uterine perfusion, the treatment effect is significantly improved.
[0106] Table 2 Treatment cure conditions of each group of dairy cows with endometritis
[0107] Group Number of test subjects / head Number of cured subjects / head Cure rate / % Example 1 27 20 74.07％ Example 3 26 21 80.77％ Example 5 28 24 85.71％ Comparative Example 1 27 4 14.81％ Comparative Example 2 27 5 18.52％ Comparative Example 3 27 10 37.04％ Comparative Example 4 27 12 44.44％ Comparative Example 5 27 17 62.96％
[0108] As can be seen from Table 2, by using the perfusion fluid and suppository of the method of the present invention, after one treatment course, the treatment cure rate of dairy cows with endometritis can rise above 75%. Compared with the effective rate of 14.81% in Comparative Example 1 where only recombinant lysostaphin was used for uterine perfusion, the treatment effect is significantly improved.
[0109] Before treatment, 5 water buffaloes with endometritis were randomly selected from each group for bacterial content detection and marked. After treatment, the marked water buffaloes were further subjected to bacterial content detection, and their average bacterial content was calculated. The results are shown in Table 3.
[0110] Table 3 Bacterial content conditions before and after treatment
[0111]
[0112]
[0113] The present invention can be implemented in various different ways and is not limited to the described embodiments. Those of ordinary skill in the art can understand that the present invention can be implemented in other specific ways without changing the technical idea or essential features of the present invention. Therefore, it should be understood that the above-described embodiments are exemplary and not intended to limit the present invention.
Claims
1. A pharmaceutical preparation for treating endometritis, characterized in that: The pharmaceutical preparation comprises a first composition and a second composition; The raw materials of the first composition include: enzyme preparation, polysorbate 80 and N-acetylcysteine; The raw materials of the second composition include: recombinant lysostaphin, lysozyme, N-acetylcysteine and a water-soluble matrix.
2. The pharmaceutical preparation for treating endometritis according to claim 1, characterized in that: The raw materials of the first composition are 15-38g of enzyme preparation, 0.1-1mL of polysorbate 80 and 1-5g of N-acetylcysteine by mass; the raw materials of the second composition are 10-20% of recombinant lysostaphin, 15-25% of lysozyme, 1-10% of N-acetylcysteine and 45-74% of water-soluble matrix by mass.
3. The pharmaceutical preparation for treating endometritis according to claim 1, characterized in that: The raw materials of the first composition also include leonurine.
4. The pharmaceutical preparation for treating endometritis according to claim 3, characterized in that: Calculated by mass, the raw materials of the first composition are 15-38 g of enzyme preparation, 0.1-1 mL of polysorbate 80, 1-5 g of N-acetylcysteine and 5-10 mg of leonurine.
5. The pharmaceutical preparation for treating endometritis according to claim 4, characterized in that: The enzyme preparation comprises one or both of Bacillus subtilis fibrinolytic enzyme and bromelain.
6. The pharmaceutical preparation for treating endometritis according to claim 1, characterized in that: The raw materials of the second composition also include: polysorbate 80, vitamin B3, polyhexanide and trehalose.
7. The pharmaceutical preparation for treating endometritis according to claim 6, characterized in that: Calculated by mass percentage, the raw materials of the second composition are 13-15% recombinant lysostaphin, 18-24% lysozyme, 1-3% polysorbate 80, 1-5% vitamin B3, 1-6% polyhexanide, 0.5-2% trehalose, 1-3% N-acetylcysteine and 35-60% glycerol gelatin.
8. A method for preparing a pharmaceutical preparation for treating endometritis, characterized in that: The preparation method comprises: S1, preparing a perfusion solution using a first composition: S11, dissolving leonurine in water to obtain a leonurine solution; dissolving Bacillus subtilis fibrinolytic enzyme in water to obtain a Bacillus subtilis fibrinolytic enzyme solution; S12, taking N-acetylcysteine, adding it to water, heating it to 25-30° C. to dissolve it, and adjusting the pH to 4.0-5.0, then adding polysorbate 80, stirring and mixing, then slowly mixing the motherwort solution and the Bacillus subtilis fibrinolytic enzyme solution into the solution, fixing the volume, and sterilizing to obtain a perfusion solution; S2, preparing a suppository using the second composition: S21, trehalose is dissolved in water, and then recombinant lysostaphin and lysozyme are added to obtain a pretreatment solution; S22, take a water-soluble matrix and heat it to 38-40°C, add polysorbate 80, vitamin B3, and polyhexanide, stir until completely dissolved, cool to 25-30°C, add the pretreatment solution and N-acetylcysteine, stir slowly to dissolve, inject into a suppository mold, and cool to 4°C to solidify.
9. The method for preparing the pharmaceutical preparation for treating endometritis according to claim 8, characterized in that: The preparation of the water-soluble matrix includes: taking gelatin, shearing it, soaking it to fully swell it, dissolving it in a 60-70° C. water bath, adding glycerol and water, stirring until it is uniform, and controlling the water content to be 10-20% to obtain a water-soluble matrix glycerol gelatin.
10. Use of a pharmaceutical preparation for treating endometritis according to any one of claims 1 to 7, characterized in that: The pharmaceutical preparation is an external medicine, and the method of use is to first use the infusion solution prepared from the first composition, and then use the suppository prepared from the second composition.