A suppository for treating bovine and ovine endometritis, a preparation method and application thereof

Abstract

This invention discloses a suppository for treating endometritis in cattle and sheep, its preparation method, and its application, belonging to the field of veterinary drug preparation technology. The suppository uses Salvia miltiorrhiza, Coptis chinensis, Forsythia suspensa, Carthamus tinctorius, Borneol, and Astragalus membranaceus as active ingredients. Coptis chinensis and Forsythia suspensa clear damp-heat, while Salvia miltiorrhiza and Carthamus tinctorius unblock stagnation. Astragalus membranaceus replenishes postpartum qi deficiency, promotes blood circulation, enhances blood circulation, strengthens the body's resistance to pathogens, prevents inflammation recurrence, harmonizes the cold properties of other herbs, and reduces side effects. Borneol, with its pungent and penetrating properties, guides the medicine directly to the lesion, promotes mucosal repair, increases mucosal permeability, relieves pain and itching, and synergistically provides anti-inflammatory and analgesic effects, improving the user experience. It is suitable for local suppository administration, aligning with the principles of traditional Chinese veterinary medicine for endometritis, simultaneously exerting the effects of clearing heat and dampness, promoting blood circulation and removing blood stasis, and tonifying qi and strengthening the body, addressing both the symptoms and the root cause. By adding a matrix and excipients and combining vaginal administration to treat endometritis in cattle and sheep, the local drug concentration is concentrated for rapid onset of action, achieving a balance between effectiveness, convenience, and safety.

Description

Technical Field

[0001] This invention belongs to the field of veterinary drug preparation technology, specifically relating to a suppository for treating endometritis in cattle and sheep, its preparation method, and its application. Background Technology

[0002] In large-scale, intensive cattle and sheep farming, endometritis is a prevalent reproductive system disease in postpartum cattle and sheep, with an incidence rate of 20% to 40%. The core contributing factors include postpartum uterine mucosal damage, pathogenic bacterial infections (such as Escherichia coli and Staphylococcus aureus), and decreased immunity. This disease not only disrupts estrus and reduces conception rates in cattle and sheep, but in severe cases can lead to pyometra and even infertility, causing significant economic losses to the livestock industry. Therefore, developing safe, effective, and suitable treatment drugs and administration methods for cattle and sheep farming is a crucial requirement for ensuring reproductive performance and farming efficiency.

[0003] Currently, intrauterine drug administration requires professional personnel to use vaginal speculum and catheters, which can easily cause stress and struggle in animals and may also damage the mucosa. These problems increase the difficulty of developing drugs for the treatment of endometritis. Clinically, drugs and administration techniques used to treat endometritis in cattle and sheep have formed a diverse system, mainly divided into three categories: First, antibiotics such as gentamicin sulfate and enrofloxacin, which are mostly administered via uterine injection to achieve antibacterial and anti-inflammatory effects; second, traditional Chinese medicine preparations, including oral compound preparations such as Yimu Sheng Hua San and local infusion preparations such as Danlianhua uterine infusion solution, which rely on their effects of clearing heat and promoting blood circulation, antibacterial and repairing; and third, suppository preparations, such as traditional Chinese medicine compound suppositories containing ingredients like Coptis chinensis and Sophora flavescens, which are administered locally via intrauterine delivery. Among these, traditional Chinese medicine preparations have gained increasing attention and application in the field of veterinary drug preparation due to their high safety, ability to regulate bodily functions through different pathways, and alignment with the trends of green and antibiotic-free farming. For example, Chinese patent application CN116236524A discloses a method for preparing a uterine irrigation solution containing Coptis chinensis, Forsythia suspensa, Salvia miltiorrhiza, and Carthamus tinctorius. The solution is obtained by soaking, reflux extraction, and concentration of four medicinal materials: Coptis chinensis, Forsythia suspensa, Salvia miltiorrhiza, and Carthamus tinctorius. It has broad-spectrum antibacterial, anti-inflammatory, and blood-activating properties and has shown certain application value in the treatment of endometritis in dairy cows.

[0004] While existing drugs and administration methods for treating endometritis in cattle and sheep can play a certain therapeutic role, they still have the following limitations: (1) The inherent limitations of Danlianhua uterine irrigation solution. As a local treatment preparation of traditional Chinese medicine, Danlianhua uterine irrigation solution has advantages such as no drug resistance and no milk withdrawal period, but its irrigation formulation has unavoidable defects in cattle and sheep use: including high threshold for administration and strong stress: irrigation administration requires exposing the cervix and inserting a catheter through a vaginal speculum. The operation process needs to be completed by professionals, and the insertion of instruments can easily cause cattle and sheep to have a strong stress response, causing them to struggle and strain. This not only increases the difficulty of operation, but may also cause secondary damage to the vaginal mucosa, and even aggravate the risk of inflammation spread. For large-scale cattle and sheep farms, this operation mode is time-consuming and laborious, and it is difficult to adapt to the high-efficiency treatment needs of a large number of sick cattle and sheep. Low drug utilization and easy loss: After the infusion solution is administered, the drug solution is easy to flow out with the uterine secretions when the cattle and sheep stand or move, which leads to a rapid decrease in the drug concentration at the lesion site. It is necessary to increase the dosage and frequency of administration to maintain the therapeutic effect, which not only wastes the drug materials, but also prolongs the treatment cycle. Insufficient storage and transportation convenience: Liquid dosage forms have high requirements for packaging sealing and are prone to precipitation and stratification in low temperature environments, affecting drug stability; at the same time, the liquid form increases the risk of leakage during transportation, which is not conducive to the supply of drugs in remote breeding areas. (2) Core defects of antibiotic drugs. Antibiotics represented by gentamicin sulfate and enrofloxacin are commonly used clinical treatment drugs, but they have multiple problems: including prominent drug resistance and residue risks: long-term use of single or combined antibiotics can easily induce drug resistance in pathogens, but long-term use will lead to the spread of drug-resistant strains, and the subsequent treatment effect will be significantly reduced. At the same time, antibiotics are easy to form residues in beef and mutton and beef and mutton milk, which does not meet food safety standards. It is necessary to strictly implement the withdrawal period, which affects the economic benefits of breeding. There are shortcomings in the administration methods: injection administration requires professional operation, and multiple injections will increase the stress and infection risk of cattle and sheep. Moreover, the drug needs to be metabolized throughout the body before reaching the lesion, resulting in insufficient local concentration and slow onset of action. Uterine injection administration faces similar complex operation and drug loss problems as Danlianhua irrigation solution. Furthermore, the killing of normal flora in the uterus by antibiotics can easily disrupt the microecological balance. (3) The compatibility of existing Chinese medicine preparations and suppositories is insufficient. The efficacy of oral Chinese medicine compound prescriptions is limited: oral Chinese medicines such as Yimu Shenghua San need to be digested and absorbed through the rumen. The acid-base environment and microorganisms in the rumen can easily destroy heat-sensitive active ingredients such as berberine and chlorogenic acid, leading to a decrease in efficacy. At the same time, the drug needs to reach the uterine lesion through blood circulation, resulting in low local drug concentration, slow onset of action, and a burden on the gastrointestinal tract of postpartum cattle and sheep with weak digestive system function.Existing suppositories have several limitations in their application scenarios: While published compound suppositories for livestock (such as the Chinese patent application with publication number CN101879240A) have solved the stability problem of liquid dosage forms, they still have significant shortcomings in terms of application: First, the formulations are designed for dairy cows and do not take into account the physiological characteristics of sheep, such as their small weight and limited uterine volume. The dosage (usually several grams per sheep based on the amount of raw materials) far exceeds the tolerance range of sheep. Second, the administration method is intrauterine delivery, which still requires professional operation and does not take advantage of the convenience of vaginal administration. Third, some suppositories need to be dissolved in physiological saline before use, which is essentially still an infusion mode and does not achieve fundamental optimization of the dosage form. Fourth, the suppository matrix has poor drug release performance or is prone to melting at temperatures above 37°C (especially in some areas where summer temperatures are high, the suppositories melt directly inside the packaging and cannot be administered).

[0005] In summary, existing drugs for treating endometritis generally suffer from a trade-off between ease of administration and local efficacy: while oral and injection administration are relatively standardized, it is difficult for the drug to achieve an effective concentration at the uterine lesion; uterine instillation and traditional intrauterine suppositories can achieve local effects, but the procedures are complex, highly irritating, and prone to drug loss. Furthermore, the contradiction between insufficient dosage leading to poor efficacy and excessive dosage increasing the burden on the patient, fails to meet the core requirements of high efficiency, convenience, and safety in large-scale farming. Summary of the Invention

[0006] In order to overcome the shortcomings of the prior art, the present invention aims to provide a suppository for treating endometritis in cattle and sheep, its preparation method and application, so as to solve the technical problem that existing endometritis treatment drugs cannot simultaneously achieve high efficiency, convenience and safety.

[0007] To achieve the above objectives, the present invention employs the following technical solution: The first aspect of the present invention discloses a suppository for treating endometritis in cattle and sheep, comprising salvia miltiorrhiza, coptis chinensis, forsythia suspensa, safflower, borneol, astragalus membranaceus, a matrix and excipients in a mass ratio of (3~5):(10~20):(10~20):(2~8):(2~8):(3~5):(25~45):(15~30).

[0008] Preferably, the matrix comprises polyethylene glycol and cetearyl alcohol.

[0009] More preferably, the mass ratio of polyethylene glycol to cetearyl alcohol is (10~40):(30~60).

[0010] Preferably, the excipients include sodium carboxymethyl cellulose, carbomer, sodium alginate, and glycerin.

[0011] More preferably, the mass ratio of sodium carboxymethyl cellulose, carbomer, sodium alginate and glycerin is (5~15):(5~20):(8~10):(2~5).

[0012] More preferably, the mass ratio of sodium carboxymethyl cellulose, carbomer, sodium alginate and glycerin is (8~12):(10~15):(8~10):(2~5).

[0013] In a second aspect, this invention discloses a method for preparing a suppository for treating endometritis in cattle and sheep. The method involves mixing Salvia miltiorrhiza, Coptis chinensis, Forsythia suspensa, Carthamus tinctorius, Borneol, and Astragalus membranaceus, then soaking the mixture in water at 20 times the total weight of the herbs. The mixture is then decocted twice at 90-100°C, 1.5 hours each time. The decoctions are combined, filtered, precipitated with alcohol, concentrated, and then a matrix and excipients are added and melted. The mixture is then molded to obtain the suppository for treating endometritis in cattle and sheep.

[0014] Preferably, the alcohol precipitation step is as follows: add ethanol to the filtrate to make the ethanol concentration reach 70%, stir evenly, refrigerate at 4°C and let stand, filter and take the supernatant.

[0015] Preferably, the concentration step is as follows: the supernatant after alcohol precipitation is concentrated under reduced pressure at 60~65℃ to a volume of 1 / 100 of the original filtrate.

[0016] A third aspect of the present invention discloses the use of a suppository for treating bovine and ovine endometritis in the preparation of a drug for treating bovine and ovine endometritis.

[0017] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a suppository for treating endometritis in cattle and sheep. 1) In the formula, Coptis chinensis and Forsythia suspensa clear damp-heat, while Salvia miltiorrhiza and Carthamus tinctorius unblock stagnation; Astragalus membranaceus replenishes postpartum qi deficiency, promotes blood circulation, enhances blood circulation, removes blood stasis, strengthens the body's resistance to pathogens, prevents recurrence of inflammation, harmonizes the cold properties of the herbs, and reduces side effects; Borneol is pungent and aromatic, guiding the medicine directly to the lesion, promoting mucosal repair, increasing mucosal permeability, relieving pain and itching, synergistically anti-inflammatory and analgesic, improving the user experience, and meeting the needs of local suppository administration. This formula aligns with the pathogenesis of endometritis in traditional Chinese veterinary medicine (damp-heat, blood stasis, and qi deficiency), simultaneously exerting the effects of clearing heat and removing dampness, promoting blood circulation and removing blood stasis, and tonifying qi and strengthening the body, addressing both the symptoms and the root cause. 2) The vaginal suppository formulation eliminates the need for complex equipment, allowing ordinary farmers to easily administer the medication and avoiding secondary injuries from instrument insertion. This significantly reduces stress in cattle and sheep, meeting the needs of large-scale cattle and sheep farms for batch treatment. It addresses the drawbacks of traditional infusion solutions, such as high operational barriers and strong stress. Furthermore, it facilitates storage and transportation, eliminating concerns about leakage or low-temperature precipitation, thus overcoming the inconvenience of storing and transporting infusion solutions and the poor compatibility and instability of existing suppositories. Therefore, this suppository for treating endometritis in cattle and sheep combines the active ingredients (Salvia miltiorrhiza, Coptis chinensis, Forsythia suspensa, Carthamus tinctorius, Borneol, and Astragalus membranaceus), matrix, and excipients with vaginal administration. Through localized drug concentration enrichment, it achieves rapid onset of action, reducing the impact of the drug on the rumen microecology and systemic organs, thus balancing the effectiveness, convenience, and safety of treating endometritis in cattle and sheep. Experiments have shown that this suppository can maintain its efficacy for up to 3 days, and animals with endometritis only need 1-2 doses for a cure. Compared to conventional formulations, it significantly reduces the workload for farmers and minimizes the stress on animals during administration.

[0018] Furthermore, using polyethylene glycol and cetearyl alcohol as the matrix ensures that the suppository does not melt at a high temperature of 50°C. After being inserted into the vagina, it is slowly absorbed and released. When it comes into contact with vaginal secretions, it dissolves to form a viscous gel that remains in the vagina and is slowly absorbed into the uterus.

[0019] Furthermore, by adding sodium carboxymethyl cellulose, carbomer, sodium alginate, and glycerin as excipients to the matrix, the release of drugs from the suppositories is slowed down, the vaginal mucosa is not irritated, the duration of drug efficacy is prolonged, and the frequency of administration is reduced.

[0020] Furthermore, by adjusting the ratio of polyethylene glycol and glycerin in the matrix, the absorption of the drug through the vaginal mucosa can be promoted, allowing it to exert its therapeutic effect inside the uterus.

[0021] This invention provides a method for preparing suppositories for treating endometritis in cattle and sheep. Firstly, during water extraction, 20 times the volume of purified water is used, and the suppositories are boiled at 90-100°C, combined with ethanol precipitation, to more fully extract the active ingredients and remove impurities. Secondly, concentration allows for a high concentration of active ingredients, and the suppositories slowly dissolve and release within the vagina, minimizing drug loss. The precise dosage of 25g per suppository, containing 10g of raw medicinal materials, is tailored to the physiological structure of the cattle and sheep vagina, eliminating the need for additional dosage. Compared to irrigation solutions, this method reduces material waste, lowers breeding costs, and solves the problems of low drug utilization and easy loss. Thirdly, precise dosage combined with weight variation control within ±5% ensures accurate dosage for each animal, avoiding insufficient or excessive dosage. Furthermore, the raw medicinal materials meet the standards of the Chinese Pharmacopoeia, are free from mold and insect infestation, and the process involves no harmful additives, ensuring safe use and complying with food safety regulations in the livestock industry, while mitigating the risks of antibiotic resistance and residues. Attached Figure Description

[0022] Figure 1 This is a photograph of the suppository for treating endometritis in cattle and sheep according to the present invention. Figure 2 This is a comparative chart showing the uterine health of cattle and sheep after treating endometritis with different formulations of the present invention. Detailed Implementation

[0023] To enable those skilled in the art to understand the features and effects of the present invention, the following description and definitions are only general descriptions of the terms and expressions mentioned in the specification. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meaning understood by those skilled in the art regarding the present invention, and in case of conflict, the definitions in this specification shall prevail.

[0024] The theories or mechanisms described and disclosed herein, whether right or wrong, should not in any way limit the scope of the invention, that is, the contents of the invention can be implemented without being limited by any particular theory or mechanism.

[0025] In this document, all features defined by numerical ranges or percentage ranges, such as numerical values, quantities, contents, and concentrations, are for the sake of brevity and convenience only. Accordingly, descriptions of numerical ranges or percentage ranges should be considered as covering and specifically disclosing all possible sub-ranges and individual numerical values ​​(including integers and fractions) within those ranges.

[0026] In this article, unless otherwise specified, “contains,” “includes,” “containing,” “has,” or similar terms cover the meanings of “composed of” and “mainly composed of,” for example, “A contains a” covers the meanings of “A contains a and others” and “A contains only a.”

[0027] For the sake of brevity, not all possible combinations of the technical features in each implementation scheme or embodiment are described herein. Therefore, as long as there is no contradiction in the combination of these technical features, the technical features in each implementation scheme or embodiment can be combined arbitrarily, and all possible combinations should be considered within the scope of this specification.

[0028] In this article, "room temperature" refers to a temperature of approximately 20°C to 35°C, or approximately 23°C to 28°C, or approximately 25°C. It can be 20°C, 25°C, 30°C, or 33°C.

[0029] This invention provides a suppository for treating endometritis in cattle and sheep, comprising Salvia miltiorrhiza, Coptis chinensis, Forsythia suspensa, Carthamus tinctorius, Borneol, Astragalus membranaceus, a matrix and excipients in a mass ratio of (3~5):(10~20):(10~20):(2~8):(2~8):(3~5):(25~45):(15~30); The matrix comprises polyethylene glycol and cetearyl alcohol in a mass ratio of (10~40):(30~60); the excipients comprise sodium carboxymethyl cellulose, carbomer, sodium alginate and glycerin in a mass ratio of (5~15):(5~20):(8~10):(2~5); the preferred mass ratio of polyethylene glycol and cetearyl alcohol is (20~30):(40~50); the preferred mass ratio of sodium carboxymethyl cellulose, carbomer, sodium alginate and glycerin is (8~12):(10~15):(8~10):(2~5).

[0030] This invention provides a method for preparing a suppository for treating endometritis in cattle and sheep, comprising the following steps: 1. Water extraction: Mix the herbs Salvia miltiorrhiza, Coptis chinensis, Forsythia suspensa, Carthamus tinctorius, Borneol and Astragalus membranaceus in a mass ratio of (3~5):(10~20):(10~20):(2~8):(2~8):(3~5), add 20 times the amount of purified water, soak for 1~4 hours, decoct twice at 90~100℃ for 1.5 hours each time, combine the decoctions and filter.

[0031] 2. Alcohol precipitation: Slowly add 95% (v / v) pharmaceutical ethanol to the filtrate obtained in step 1 to make the ethanol concentration reach 70%, stir evenly, refrigerate at 4°C and stand for 24 h, then filter and take the supernatant.

[0032] 3. Concentration: The supernatant obtained in step 2 is concentrated under reduced pressure at a temperature of 60~65℃ and a pressure of 0.06~0.09 MPa until the volume is 1 / 100 of the original filtrate, to obtain a concentrated Chinese medicine solution.

[0033] 4. Matrix melting: Based on the calculation of "each suppository contains 10 g of raw medicinal materials", take the corresponding amount of Chinese medicine concentrate, add mixed matrix and excipients (the amount of matrix is ​​4 to 5 times the mass of Chinese medicine concentrate), and stir in a water bath at 65 to 70°C until melted and clear.

[0034] 5. Injection molding: Preheat the suppository mold (25 g / pill) to 65~70℃, inject the molten medicine, cool to room temperature, demold, and obtain suppositories for treating endometritis in cattle and sheep. The weight difference of the suppositories should be within ±5%.

[0035] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading this description, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined in this application.

[0036] The following examples use instruments and equipment conventional in the art. Experimental methods in the following examples, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. The raw medicinal materials used in the following examples—Salvia miltiorrhiza, Coptis chinensis, Forsythia suspensa, Carthamus tinctorius, Borneol, and Astragalus membranaceus—all meet the standards of the Chinese Pharmacopoeia and are free from mold and insect infestation. Astragalus membranaceus was purchased from Anhui Zhang Zhongjing Traditional Chinese Medicine Technology Co., Ltd., batch number: 250404-2; Salvia miltiorrhiza was purchased from Shaanxi Bainianshou Pharmaceutical Co., Ltd., batch number: S20240801; Forsythia suspensa was purchased from Shaanxi Shangluo Panlong Plant Pharmaceutical Co., Ltd., batch number: 20240401; Borneol was purchased from Chengdu University of Traditional Chinese Medicine Qin Kang Pharmaceutical Technology Co., Ltd., batch number: 240901; Carthamus tinctorius was purchased from Shaanxi Bainianshou Pharmaceutical Co., Ltd. Product batch number: S20241001; Coptis chinensis, purchased from Shaanxi Shangluo Panlong Botanical Pharmaceutical Co., Ltd., product batch number: 20240701; the polyethylene glycol (PEG4000, melting point 40~48℃) and cetearyl alcohol (melting point 48~54℃) used are both pharmaceutical grade; polyethylene glycol was purchased from Sigma-Aldrich, catalog number 95904; cetearyl alcohol was purchased from Sigma-Aldrich, catalog number 68240-500G-F; all other raw materials used, unless otherwise specified, are conventional commercially available products with specifications that are standard in the field.

[0037] I. Preparation of Suppositories for Treating Endometritis in Cattle and Oxygen Example 1 1. Water extraction Accurately weigh 4 g of Salvia miltiorrhiza, 20 g of Coptis chinensis, 15 g of Forsythia suspensa, 6 g of Carthamus tinctorius, 4 g of Borneol and 3 g of Astragalus membranaceus. Add 20 times the amount of purified water as the total amount of medicinal materials, soak for 1 hour, and then decoct twice at 100℃ for 1.5 hours each time. Combine the two decoctions and filter them under normal pressure using a double-layer 100-mesh filter cloth to obtain the filtrate.

[0038] 2. Alcohol precipitation Slowly add 95% (v / v) pharmaceutical ethanol to the filtrate obtained in step 1, while continuously stirring at 100 r / min until the ethanol concentration in the system reaches 70%. After stirring evenly, place it in a refrigerated environment at 4℃ and let it stand for 24 h. Filter again and collect the supernatant.

[0039] 3. Concentrated The supernatant obtained in step 2 was concentrated under reduced pressure at 65°C and 0.08 MPa until the volume of the concentrated liquid was 1 / 100 of the volume of the filtrate in step 1, thus obtaining the traditional Chinese medicine concentrate.

[0040] 4. Matrix melting Accurately weigh 15 g of polyethylene glycol and 25 g of cetearyl alcohol and mix them evenly to obtain a mixed matrix.

[0041] Take 10 g of the concentrated Chinese medicine solution obtained in step 3, add 40 g of mixed matrix, 9 g of sodium carboxymethyl cellulose, 10 g of carbomer, 8 g of sodium alginate and 3 g of glycerol, and place in a 70°C water bath and stir continuously until the system is completely melted and clear.

[0042] 5. Injection molding Preheat 25 ml centrifuge tubes to 70°C, then inject the molten drug solution obtained in step 4 into them at a uniform rate. Cool at room temperature (25°C) for 30 min. After the drug solution has completely solidified, demold the tubes and screen out products with a weight difference within ±5% to obtain suppositories for treating endometritis in cattle and sheep.

[0043] Example 2 The difference from Example 1 lies in the amount of matrix and excipients used. The specific steps are as follows: 1. Water extraction Accurately weigh 4 g of Salvia miltiorrhiza, 20 g of Coptis chinensis, 15 g of Forsythia suspensa, 6 g of Carthamus tinctorius, 4 g of Borneol and 3 g of Astragalus membranaceus. Add 20 times the amount of purified water as the total amount of medicinal materials, soak for 1 hour, and then decoct twice at 100℃ for 1.5 hours each time. Combine the two decoctions and filter them under normal pressure using a double-layer 100-mesh filter cloth to obtain the filtrate.

[0044] 2. Alcohol precipitation Slowly add 95% (v / v) pharmaceutical ethanol to the filtrate obtained in step 1, while continuously stirring at 100 r / min until the ethanol concentration in the system reaches 70%. After stirring evenly, place it in a refrigerated environment at 4℃ and let it stand for 24 h. Filter again and collect the supernatant.

[0045] 3. Concentrated The supernatant obtained in step 2 was concentrated under reduced pressure at 65°C and 0.08 MPa until the volume of the concentrated liquid was 1 / 100 of the volume of the filtrate in step 1, thus obtaining the traditional Chinese medicine concentrate.

[0046] 4. Matrix melting Accurately weigh 20 g of polyethylene glycol and 20 g of cetearyl alcohol and mix them evenly to obtain a mixed matrix.

[0047] Take 10 g of the concentrated Chinese medicine solution obtained in step 3, add 40 g of mixed matrix, 7 g of sodium carboxymethyl cellulose, 5 g of carbomer, 9 g of sodium alginate and 2 g of glycerol, and place in a 70°C water bath and stir continuously until the system is completely melted and clear.

[0048] 5. Injection molding Preheat 25 ml centrifuge tubes to 70°C, then inject the molten drug solution obtained in step 4 into them at a uniform rate. Cool at room temperature (25°C) for 30 min. After the drug solution has completely solidified, demold the tubes and screen out products with a weight difference within ±5% to obtain suppositories for treating endometritis in cattle and sheep.

[0049] Example 3 The difference from Example 1 lies in the amount of matrix and excipients used. The specific steps are as follows: 1. Water extraction Accurately weigh 4 g of Salvia miltiorrhiza, 20 g of Coptis chinensis, 15 g of Forsythia suspensa, 6 g of Carthamus tinctorius, 4 g of Borneol and 3 g of Astragalus membranaceus. Add 20 times the amount of purified water as the total amount of medicinal materials, soak for 1 hour, and then decoct twice at 100℃ for 1.5 hours each time. Combine the two decoctions and filter them under normal pressure using a double-layer 100-mesh filter cloth to obtain the filtrate.

[0050] 2. Alcohol precipitation Slowly add 95% (v / v) pharmaceutical ethanol to the filtrate obtained in step 1, while continuously stirring at 100 r / min until the ethanol concentration in the system reaches 70%. After stirring evenly, place it in a refrigerated environment at 4℃ and let it stand for 24 h. Filter again and collect the supernatant.

[0051] 3. Concentrated The supernatant obtained in step 2 was concentrated under reduced pressure at 65°C and 0.08 MPa until the volume of the concentrated liquid was 1 / 100 of the volume of the filtrate in step 1, thus obtaining the traditional Chinese medicine concentrate.

[0052] 4. Matrix melting Accurately weigh 10 g of polyethylene glycol and 30 g of cetearyl alcohol and mix them evenly to obtain a mixed matrix.

[0053] Take 10 g of the concentrated Chinese medicine solution obtained in step 3, add 40 g of mixed matrix, 5 g of sodium carboxymethyl cellulose, 10 g of carbomer, 8 g of sodium alginate and 5 g of glycerol, and place in a 70°C water bath and stir continuously until the system is completely melted and clear.

[0054] 5. Injection molding Preheat 25 ml centrifuge tubes to 70°C, then inject the molten drug solution obtained in step 4 into them at a uniform rate. Cool at room temperature (25°C) for 30 min. After the drug solution has completely solidified, demold the tubes and screen out products with a weight difference within ±5% to obtain suppositories for treating endometritis in cattle and sheep.

[0055] Comparative Example 1 The difference from Example 1 is that it does not contain Astragalus membranaceus. The specific steps are as follows: 1. Water extraction Accurately weigh 4 g of Salvia miltiorrhiza, 20 g of Coptis chinensis, 15 g of Forsythia suspensa, 6 g of Carthamus tinctorius, and 4 g of Borneol. Add 20 times the amount of purified water as the total medicinal material, soak for 1 hour, and then decoct twice at 100°C for 1.5 hours each time. Combine the two decoctions and filter them under normal pressure using a double-layer 100-mesh filter cloth to obtain the filtrate.

[0056] 2. Alcohol precipitation Slowly add 95% (v / v) pharmaceutical ethanol to the filtrate obtained in step 1, while continuously stirring at 100 r / min until the ethanol concentration in the system reaches 70%. After stirring evenly, place it in a refrigerated environment at 4℃ and let it stand for 24 h. Filter again and collect the supernatant.

[0057] 3. Concentrated The supernatant obtained in step 2 was concentrated under reduced pressure at 65°C and 0.08 MPa until the volume of the concentrated liquid was 1 / 100 of the volume of the filtrate in step 1, thus obtaining the traditional Chinese medicine concentrate.

[0058] 4. Matrix melting Accurately weigh 15 g of polyethylene glycol and 25 g of cetearyl alcohol and mix them evenly to obtain a mixed matrix.

[0059] Take 10 g of the concentrated Chinese medicine solution obtained in step 3, add 40 g of mixed matrix, 9 g of sodium carboxymethyl cellulose, 10 g of carbomer, 8 g of sodium alginate and 3 g of glycerol, and place in a 70°C water bath and stir continuously until the system is completely melted and clear.

[0060] 5. Injection molding Preheat 25 ml centrifuge tubes to 70°C, then inject the molten drug solution obtained in step 4 into them at a uniform rate. Cool at room temperature (25°C) for 30 min. After the drug solution has completely solidified, demold the tubes and screen out products with a weight difference within ±5% to obtain suppositories without Astragalus membranaceus.

[0061] Comparative Example 2 The difference from Example 1 is that it does not contain borneol and astragalus. The specific steps are as follows: 1. Water extraction Accurately weigh 4 g of Salvia miltiorrhiza, 20 g of Coptis chinensis, 15 g of Forsythia suspensa and 6 g of Carthamus tinctorius. Add 20 times the amount of purified water as the total amount of medicinal materials, soak for 1 hour, and then decoct twice at 100℃ for 1.5 hours each time. Combine the two decoctions and filter them under normal pressure using a double-layer 100-mesh filter cloth to obtain the filtrate.

[0062] 2. Alcohol precipitation Slowly add 95% (v / v) pharmaceutical ethanol to the filtrate obtained in step 1, while continuously stirring at 100 r / min until the ethanol concentration in the system reaches 70%. After stirring evenly, place it in a refrigerated environment at 4℃ and let it stand for 24 h. Filter again and collect the supernatant.

[0063] 3. Concentrated The supernatant obtained in step 2 was concentrated under reduced pressure at 65°C and 0.08 MPa until the volume of the concentrated liquid was 1 / 100 of the volume of the filtrate in step 1, thus obtaining the traditional Chinese medicine concentrate.

[0064] 4. Matrix melting Accurately weigh 15 g of polyethylene glycol and 25 g of cetearyl alcohol and mix them evenly to obtain a mixed matrix.

[0065] Take 10 g of the concentrated Chinese medicine solution obtained in step 3, add 40 g of mixed matrix, 9 g of sodium carboxymethyl cellulose, 10 g of carbomer, 8 g of sodium alginate and 3 g of glycerol, and place in a 70°C water bath and stir continuously until the system is completely melted and clear.

[0066] 5. Injection molding Preheat 25 ml centrifuge tubes to 70°C, then inject the molten drug solution obtained in step 4 into them at a uniform rate. Cool at room temperature (25°C) for 30 min. After the drug solution has completely solidified, demold the tubes and screen out products with a weight difference within ±5% to obtain suppositories that do not contain borneol or astragalus.

[0067] II. Performance Evaluation of Suppositories for Treating Endometritis in Bovine and Ovine Women 1. Observation of appearance Take 3 suppositories prepared in Example 1 for treating endometritis in cattle and sheep, and visually observe their color, shape, and texture under natural light and at room temperature (25°C), and record them.

[0068] The results are as follows Figure 1 As shown, the suppositories for treating endometritis in cattle and sheep are brownish-black, hard, intact in shape, smooth in surface, and free from cracks, deformation, and obvious impurities.

[0069] 2. Determination of the melting time of suppositories Referring to the "Melt-out Time Test Method" in the Veterinary Pharmacopoeia of the People's Republic of China, the basket method was used to determine the melt-out time of suppositories.

[0070] Take three suppositories prepared in Example 1 for treating bovine and ovine endometritis and place them in three metal baskets. Immerse the baskets in beakers containing purified water at a constant temperature of 37±0.5℃, with the water level above the top of the baskets. Start the instrument and start timing. Observe and record the time from the beginning of melting to complete disintegration, dissolution, or detachment of the suppository and passing through the sieve. This is the melting time limit, and the average value of the three suppositories is taken.

[0071] The results showed that the melting time of the suppository for treating endometritis in cattle and sheep prepared in Example 1 was 4 hours.

[0072] 3. Determination of complete in vitro dissolution time of suppositories One suppository each from Examples 1, 2, and 3 prepared for treating bovine and ovine endometritis was accurately weighed and placed in a beaker containing 200 mL of constant-temperature physiological saline at 37±0.5℃. The beaker was kept in a constant-temperature water bath and stirred at 50 r / min to simulate the in vivo environment. Timing was started from the moment of immersion, and observations were made at regular intervals until the suppository was completely dissolved with no visible solid particles remaining. The time taken was recorded as the complete dissolution time. Three parallel measurements were performed, and the average value was taken.

[0073] The results showed that the complete dissolution time of the suppository for treating endometritis in cattle and sheep prepared in Example 1 was 56 h; the complete dissolution time of the suppository for treating endometritis in cattle and sheep in Example 2 was 88 h; and the complete dissolution time of the suppository for treating endometritis in cattle and sheep in Example 3 was 40 h. This result indicates that the matrix and excipient ratios in Example 1 (the mass ratio of polyethylene glycol to cetearyl alcohol in the range of (20~30):(40~50); and the mass ratio of sodium carboxymethyl cellulose, carbomer, sodium alginate, and glycerin in the range of (8~12):(10~15):(8~10):(2~5)) were optimal.

[0074] 4. Microbial limit test of suppositories 1) Preparation of test solution: Take 10 g of the suppository for treating bovine and ovine endometritis prepared in Example 1, place it in a sterile homogenizing cup containing 90 mL of 0.9% sterile sodium chloride-peptone buffer (pH 7.0), homogenize at 3000–4000 r / min for 2 min to prepare a 1:10 (g / mL) test solution, as 10 g of the solution. -1 Dilution levels, serial dilution, to obtain a concentration of 10. -1 10 -2 10 -3 The test solution.

[0075] 2) Determination of total aerobic bacteria count: Take 10 -1 10 -2 10 -3 Administer 1 mL of each dilution of the test solution to sterile Petri dishes. Immediately pour in tryptic soy agar (TSA) at approximately 45°C, shake well, and invert after solidification. Bacterial group: Incubate at 35°C for 4 days, observing and counting the number of typical colonies on the plates daily. Calculate the total aerobic bacterial count (cfu / g) based on the colony count of the appropriate dilution.

[0076] The results showed that the total number of aerobic bacteria in the suppositories for treating bovine and ovine endometritis in Example 1 was ≤10. 2 cfu / g.

[0077] 3) Determination of total mold and yeast count: Take 10 of the above-mentioned samples... -1 10 -2 Administer 1 mL of each dilution of the test solution to sterile Petri dishes, immediately pour in Sabouraud dextrose agar (SDA) at approximately 45°C, shake well, and invert after solidification. For the mold / yeast group: incubate at 25°C for 6 days, observing and counting typical colonies daily. Calculate the total number of mold and yeast cells (cfu / g) based on the colony count of the appropriate dilution plate.

[0078] The results showed that the total number of molds and yeasts in the suppositories for treating bovine and ovine endometritis in Example 1 was ≤10. 1 cfu / g.

[0079] 4) Take 10 -1 The diluted test solutions were used to perform enrichment, isolation culture, and biochemical / serological identification of Staphylococcus aureus, Salmonella, Pseudomonas aeruginosa, Candida albicans, and Clostridium.

[0080] The results showed that Staphylococcus aureus, Salmonella, Pseudomonas aeruginosa, Candida albicans, and Clostridium were not detected in the suppositories for treating endometritis in cattle and sheep in Example 1.

[0081] 5. Determination of the effective components of suppositories 1) Determination of berberine hydrochloride content: A Diamonsil C18 column was used with 0.05 mol / L potassium dihydrogen phosphate solution (40:60) as the mobile phase, column temperature 30℃, flow rate 1 mL / min, detection wavelength 345 nm, and injection volume 10 μL.

[0082] The results showed that the linear equation for the determination of berberine hydrochloride content was Y=38553X-89683, r=0.9998, and the peak area had a good linear relationship with the injection volume in the concentration range of 4.1616 to 83.232 μg / mL.

[0083] 2) Determination of the content of salvianolic acid B: A Diamonsil C18 (5 μm, 150×4.6 mm) column was used with methanol acetonitrile-formic acid-water (30:10:1:59) as the mobile phase, column temperature 30℃, flow rate 1 mL / min, detection wavelength 286 nm, and injection volume 10 μL.

[0084] The results showed that the linear equation for the determination of salvianolic acid B content was Y=1×107X-80303, r=0.9999, and the peak area had a good linear relationship with the injection volume in the concentration range of 0.02952~1.476 mg / mL.

[0085] 6. Testing the therapeutic effect of suppositories on endometritis 1) Screening of sheep with endometritis Goats clinically suspected of having endometritis were selected as candidate samples. All candidate goats came from the same breeding base, with consistent age, weight, and parity (1.5-2 years old, 45-60 kg, all multiparous (1-2 litters), and free from other reproductive system diseases and systemic infections). The breeding environment and feeding standards were uniform. Goats underwent acclimatization for 7 days prior to the experiment, with daily observation of their mental state, feed intake, and defecation. Individuals exhibiting stress reactions or latent diseases were excluded to ensure a consistent baseline. Candidate goats that passed the acclimatization were individually restrained, and their vulva was exposed using aseptic techniques. The color, texture, and transparency of vulvar discharge were observed, along with the presence of abnormal reproductive system behaviors such as arching of the back, straining, and frequent urination. Individuals with cloudy vulvar discharge (preferably red to reddish-brown) and with or without abnormal reproductive system behaviors were initially screened as suspected endometritis goats. Healthy individuals with clear or semi-clear discharge and no abnormal behavior were excluded (reserved as a subsequent control group). Thorough disinfection of the vulva and perianal area was performed on suspected diseased sheep. Uterine mucus samples were then collected from each sheep. The color and turbidity of the mucus were observed, and the percentage of polymorphonuclear leukocytes (PMN%) was measured. Sheep clinically diagnosed with endometritis were those meeting both criteria: "turbid uterine mucus, red to reddish-brown" and "PMN% > 5%". Sheep meeting only one criterion or neither were excluded from the experimental disease sample group. Healthy reserve sheep meeting both criteria—"clear or translucent uterine mucus" and "PMN% ≤ 5%"—served as the control group.

[0086] 2) Experimental Grouping Sheep clinically diagnosed with endometritis and selected healthy sheep were randomly divided into 5 groups of 6 sheep each: Control group: Healthy sheep were given only an equal volume of physiological saline and did not undergo modeling or treatment.

[0087] Endometritis group: negative control.

[0088] Treatment Group 1 (Danshen-Huanglian-Lianqiao-Honghua): Sheep with endometritis were given one vaginal suppository daily, prepared in Comparative Group 2, without borneol and Astragalus.

[0089] Treatment Group 2 (Danshen-Huanglian-Lianqiao-Honghua-Bingpian): Sheep with endometritis were given one suppository without Astragalus prepared in Comparative Group 1 via vaginal administration daily.

[0090] Treatment Group 3 (Danshen-Huanglian-Lianqiao-Honghua-Bingpian-Huangqi): Sheep with endometritis were given one vaginal suppository prepared in Example 1 daily for the treatment of endometritis in cattle and sheep.

[0091] All treatment groups received the medication for 7 consecutive days, and the clinical symptoms of the sheep were observed daily during the treatment period.

[0092] 3) Detection methods Polymorphonuclear neutrophils (PMN) percentage detection in uterine secretions: 24 hours after the last administration, a sterile swab was inserted 10-15 cm into the cervix and rotated to collect secretions. The swab was then placed in a centrifuge tube containing 1 mL of sterile saline to thoroughly wash and prepare a cell suspension. 50 μL of the cell suspension was smeared evenly, allowed to air dry, and then stained with Wright-Giemsa. 200 white blood cells were counted under a light microscope (1000× oil immersion) to calculate the percentage of neutrophils (PMN%). A PMN% > 5% in uterine secretions indicates uterine inflammation; a higher PMN% suggests a more severe local inflammatory response.

[0093] Histopathological examination: 24 hours after the last administration, endometrial tissue was collected, fixed with 4% paraformaldehyde, dehydrated with graded ethanol, embedded in paraffin, and then sectioned into 4 μm sections for HE staining. The histological structure, degree of inflammatory cell infiltration, and congestion and edema of the endometrium were observed under an optical microscope (200×, 400×).

[0094] Table 1. PMN% of vaginal secretions after drug treatment in each group

[0095] The PMN% detection results for each group are shown in Table 1, and the pathological histological detection results are as follows: Figure 2 As shown in the table, the endometritis group exhibited extensive neutrophil infiltration, congestion, and severe uterine tissue fibrosis. Compared to the endometritis group, after treatment with suppositories without borneol and astragalus, the PMN% of ovine vaginal secretions decreased to 27.42±8.41%, indicating a reduction in uterine inflammation (Table 1). While neutrophil infiltration was reduced and congestion in the uterine tissue disappeared compared to the endometritis group, severe uterine tissue fibrosis remained. Figure 2 Compared to the endometritis group, after treatment with suppositories without Astragalus membranaceus, the PMN% of ovine vaginal secretions decreased to 4.71±2.51%, indicating that the uterine inflammation was basically cured (Table 1). Compared to the endometritis group, there was no neutrophil infiltration, the uterine tissue congestion disappeared, but fibrosis still existed in the uterine tissue, with a moderate degree of fibrosis. Figure 2 Compared to the endometritis group, after treatment with the suppositories for treating bovine and ovine endometritis prepared in Example 1, the PMN% of ovine vaginal secretions decreased to 3.15±1.75% (Table 1). Compared to the endometritis group, there was no neutrophil infiltration, indicating that the inflammation in the uterus completely disappeared, the congestion in the uterine tissue disappeared, and the fibrosis in the uterine tissue disappeared. Figure 2 Meanwhile, no adverse reactions or mucosal irritation were observed in the animals after administration of the drug.

[0096] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of this invention.

Claims

1. A suppository for treating endometritis in cattle and sheep, characterized in that, The ingredients include Salvia miltiorrhiza, Coptis chinensis, Forsythia suspensa, Carthamus tinctorius, Borneol, Astragalus membranaceus, matrix and excipients in a mass ratio of (3~5):(10~20):(10~20):(2~8):(2~8):(3~5):(25~45):(15~30).

2. The suppository for treating endometritis in cattle and sheep according to claim 1, characterized in that, The matrix comprises polyethylene glycol and cetearyl alcohol.

3. The suppository for treating endometritis in cattle and sheep according to claim 2, characterized in that, The mass ratio of polyethylene glycol to cetearyl alcohol is (10~40):(30~60).

4. A suppository for treating endometritis in cattle and sheep according to any one of claims 1 to 3, characterized in that, The excipients include sodium carboxymethyl cellulose, carbomer, sodium alginate, and glycerin.

5. A suppository for treating endometritis in cattle and sheep according to claim 4, characterized in that, The mass ratio of sodium carboxymethyl cellulose, carbomer, sodium alginate and glycerin is (5~15):(5~20):(8~10):(2~5).

6. A suppository for treating endometritis in cattle and sheep according to claim 5, characterized in that, The mass ratio of sodium carboxymethyl cellulose, carbomer, sodium alginate and glycerin is (8~12):(10~15):(8~10):(2~5).

7. A method for preparing a suppository for treating endometritis in cattle and sheep according to any one of claims 1 to 6, characterized in that, After mixing the herbs Salvia miltiorrhiza, Coptis chinensis, Forsythia suspensa, Carthamus tinctorius, Borneol and Astragalus membranaceus, soak them in water with 20 times the total amount of herbs, decoct twice at 90-100℃ for 1.5 hours each time, combine the decoctions, filter, precipitate with alcohol, concentrate, add matrix and excipients to melt, and mold to obtain a suppository for treating endometritis in cattle and sheep.

8. The method for preparing a suppository for treating bovine and ovine endometritis according to claim 7, characterized in that, The steps for alcohol precipitation are as follows: add ethanol to the filtrate to make the ethanol concentration reach 70%, stir evenly, refrigerate at 4°C and let stand, then filter and take the supernatant.

9. The method for preparing a suppository for treating endometritis in cattle and sheep according to claim 7, characterized in that, The concentration steps are as follows: the supernatant after alcohol precipitation is concentrated under reduced pressure at 60~65℃ to a volume of 1 / 100 of the original filtrate.

10. The use of the suppository for treating endometritis in cattle and sheep according to any one of claims 1 to 6 in the preparation of a drug for treating endometritis in cattle and sheep.

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