Soluble powder for preventing and treating necrotic enteritis

Abstract

The invention discloses soluble powder for preventing and treating chicken necrotic enteritis, and belongs to the technical field of veterinary medicine preparations. The soluble powder preparation comprises the following components: 30-80% of penicillin V potassium, 1-20% of a beta-lactam inhibitor, 0.5-6% of a pH regulator, 0-10% of an anti-caking agent, 0.02-0.5% of a metal ion chelating agent and the balance of a filler, totaling 100%. The soluble powder is used for drinking water of poultry, and the content of penicillin V potassium in an aqueous solution of the soluble powder is still kept at 99% or above after 12 hours and is still kept at 95% or above after 24 hours. The water solution of the soluble powder does not block a water drinking line in clinical use, so that the clinical use effect and convenience are greatly improved.

Description

Technical Field

[0001] The invention belongs to the field of veterinary drug preparations, and in particular relates to a soluble powder for preventing and treating necrotic enteritis in chickens. Background Art

[0002] Necrotic enteritis in chickens is a bacterial disease caused by Clostridium perfringens (CP). Under natural infection, broilers, laying hens and native chickens can all develop the disease, and it is more common in 2 to 5 weeks of age. The disease can occur throughout the year, and the incidence rate is higher in the hot and humid summer, up to 38%, which usually leads to a decrease in the number of eggs laid by laying hens, a decline in egg quality, and an increase in the culling rate of laying hens. In recent years, as the poultry industry continues to develop in the direction of scale and intensiveness, the scale of poultry farming has gradually expanded, resulting in an increase in the incidence of necrotic enteritis in chickens, which often seriously damages the economic benefits of farmers due to the failure to take effective preventive measures in a timely manner.

[0003] Penicillin V potassium (phenoxymethyl penicillin potassium) is an oral acid-resistant semi-synthetic penicillin with the same antibacterial spectrum as penicillin G and good oral absorption. Penicillin V potassium soluble powder is available in the Netherlands and other countries for the treatment of necrotic enteritis in chickens caused by Clostridium perfringens that is sensitive to penicillin V potassium, and is used to reduce the mortality rate of chickens. Clinically proven, penicillin V potassium is very effective against necrotic enteritis in chickens caused by Clostridium perfringens. The MIC values ​​of 142 strains of Clostridium perfringens isolated from clinical cases of necrotic enteritis were experimentally determined. The MIC50 and MIC90 of penicillin V potassium are both less than 0.01μg / ml; amoxicillin MIC50 = 0.27μg / ml, MIC90 = 0.44μg / ml; erythromycin MIC50 = 0.35μg / ml, MIC90 = 0.49μg / ml; lincomycin MIC50 = 0.35μg / ml, MIC90 = 0.69μg / ml; tylosin MIC50 = 0.31μg / ml, MIC90 = 0.78μg / ml; tylosin MIC50 = 0.164μg / ml, MIC90 = 0.2748μg / ml.

[0004] With the widespread use of penicillin drugs in the market, drug-resistant strains continue to emerge. Resistance mechanisms include the production of β-lactamase, changes in target proteins, changes in cell membrane permeability, and efflux pump mechanisms. Among them, the most important mechanism for drug resistance is the production of β-lactamase. β-lactamase inhibitors can inhibit β-lactamase and prevent β-lactam antibiotics from being hydrolyzed and inactivated. However, the number of poultry farms in China is huge, and the incidence of necrotic enteritis in chickens is high. Therefore, in order to prevent the emergence of resistant strains to penicillin V potassium and improve the in vitro antibacterial effect of penicillin V potassium, it is necessary to study this. There are no reports in this regard in China.

[0005] Soluble powder is a conventional drug dosage form for poultry, which is helpful for centralized drinking water administration of poultry, easier to mix evenly, and has better effect on group disease prevention and treatment. The Chinese patent of CN 108125911 A discloses the preparation and application of a veterinary penicillin V potassium soluble powder. The patent selects to combine penicillin V potassium with an acidifier, adds a stabilizer and a carrier, so that penicillin V potassium soluble powder is not easy to degrade in water, and the effective ingredient content can be kept above 98% in water for 2 hours. When the farm actually administers drugs to poultry through drinking water, drinking water is supplied all day long. If the aqueous solution is frequently changed every day, the labor cost will be increased, which is not convenient for actual use. In addition, the drinking water used for poultry farming is from various sources, such as tap water, and is more likely to be water from the farmer's own well, which is generally used directly. The metal ions, pH value, etc. in the water may affect the stability of the aqueous solution of penicillin soluble powder, resulting in a decrease in efficacy, and the prevention and treatment effect cannot be achieved, affecting the economic benefits.

[0006] Therefore, those skilled in the art need to develop a method for preparing a drug preparation for preventing and treating necrotic enteritis in chickens that has a simple preparation process, good antibacterial effect, high stability of aqueous solution, is not prone to producing drug-resistant strains, and is easy to use in practice. Summary of the invention

[0007] The purpose of the present invention is to solve one or more deficiencies in the prior art by providing the following pharmaceutical composition for the first time and adapt to the actual use in this field. The pharmaceutical combination group comprises penicillin V potassium, EDTA disodium, and a β-lactam inhibitor, which can effectively prevent the degradation of penicillin V potassium aqueous solution and effectively prevent and treat necrotic enteritis in chickens.

[0008] To achieve the above purpose, the specific technical solution of the present invention is as follows:

[0009] The invention provides a soluble powder for preventing and treating necrotic enteritis in chickens. The soluble powder comprises penicillin V potassium, a beta-lactam inhibitor, a pH regulator, an antifreeze agent, a metal ion chelating agent and a filler.

[0010] The components and their weight percentages in the soluble powder are 30-80% of potassium penicillin V, 1-20% of beta-lactam inhibitor, 0.5-6% of pH regulator, 0-10% of anti-caking agent, 0.02-0.5% of metal ion chelating agent, and the filler is added to 100%.

[0011] Preferably, the components and their weight percentages in the soluble powder are 35-50% potassium penicillin V, 3.5-9.2% β-lactam inhibitor, 0.6-4.6% pH regulator, 2-4% anti-caking agent, 0.02-0.1% metal ion chelating agent, and the filler is added to 100%.

[0012] The β-lactam inhibitor is one or more of clavulanic acid, sulbactam, tazobactam, avibactam, ralibactam and vaborbactam.

[0013] Preferably, the β-lactam inhibitor is clavulanic acid.

[0014] The pH regulator is one or more of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, and disodium hydrogen phosphate.

[0015] The anti-caking agent is one or more of anhydrous magnesium sulfate, polyvinyl alcohol, sodium lauryl sulfate and talc.

[0016] The metal ion chelating agent is one or more of disodium EDTA, penicillamine, nitrilotriacetic acid (NTA), diisopropylaminetetraacetic acid (DTPA), and ethylenediaminetetraacetic acid (EGTA).

[0017] Preferably, the metal ion chelating agent is disodium EDTA.

[0018] The filler is one or more of maltodextrin, lactose, anhydrous glucose, β-cyclodextrin, sucrose and fructose.

[0019] Preferably, the filler is anhydrous glucose.

[0020] A more preferred formula of soluble powder for preventing and treating necrotic enteritis in chickens is: 40% potassium penicillin V, 8% clavulanic acid, 3% pH regulator, 3% anti-caking agent, 0.06% disodium EDTA, and anhydrous glucose added to 100%.

[0021] A more preferred formulation of the soluble powder for preventing and treating necrotic enteritis in chickens is: 40% potassium penicillin V, 8% clavulanic acid, 3% potassium dihydrogen phosphate, 3% anhydrous magnesium sulfate, 0.06% disodium EDTA, and anhydrous glucose added to 100%.

[0022] The present invention also provides a method for preparing a soluble powder for preventing and treating necrotic enteritis in chickens, comprising the following steps:

[0023] (1) According to the percentage by weight, potassium penicillin V, β-lactam inhibitor, pH regulator, anti-caking agent, metal ion chelating agent, and filler are respectively crushed and sieved, and the sieve mesh number is 80 mesh;

[0024] (2) The mixed materials are directly mixed to finally obtain penicillin V potassium soluble powder.

[0025] Beneficial effects:

[0026] (1) The present invention creatively uses penicillin V potassium in combination with a β-lactamase inhibitor, thereby protecting the β-lactam ring of penicillin V potassium from hydrolysis and loss of activity, and reducing the MIC value and drug resistance rate after administration. The use of a β-lactamase inhibitor increases the sensitivity of chicken Escherichia coli resistant to penicillin V potassium and enhances the antibacterial activity.

[0027] (2) The present invention uses a pH regulator to improve the stability of the soluble powder aqueous solution, and uses it in combination with a metal ion chelating agent so that the penicillin V potassium content in the aqueous solution remains above 99% after 12 hours and above 95% after 24 hours, thereby ensuring that poultry can drink water stably throughout the day.

[0028] (3) The present invention uses a highly water-soluble filler to make the soluble powder have a high water-soluble characteristic, which does not block the drinking water line during clinical use, greatly improving the effect and convenience of actual water-soluble use. At the same time, the use of an anti-caking agent ensures that the filler does not agglomerate, thereby ensuring the storage stability of the soluble powder.

[0029] (4) The penicillin V potassium soluble powder of the present invention has simple ingredients, is easy to prepare, has no special requirements for equipment, is low in cost, and is conducive to large-scale industrial production. DETAILED DESCRIPTION

[0030] The above contents will be further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter being limited to the following embodiments. All technologies realized based on the contents of the present invention are within the scope.

[0031] Example 1

[0032] A soluble powder for preventing and treating necrotic enteritis in chickens. The soluble powder comprises 35 kg of potassium penicillin V, 3.5 kg of clavulanic acid, 0.6 kg of dipotassium hydrogen phosphate, 2.0 kg of sodium lauryl sulfate, 0.02 kg of disodium EDTA, and anhydrous glucose added to 100 kg.

[0033] Preparation method:

[0034] (1) According to the percentage by weight, potassium penicillin V, clavulanic acid, dipotassium hydrogen phosphate, sodium lauryl sulfate, disodium EDTA, and anhydrous glucose are sieved separately, and the sieve mesh number is 80 mesh.

[0035] (2) The sieved materials are mixed to finally obtain penicillin V potassium soluble powder.

[0036] Example 2

[0037] A soluble powder for preventing and treating necrotic enteritis in chickens. The soluble powder comprises 40 kg of potassium penicillin V, 8 kg of clavulanic acid, 3.0 kg of potassium dihydrogen phosphate, 3.0 kg of anhydrous magnesium sulfate, 0.06 kg of disodium EDTA, and anhydrous glucose added to 100 kg.

[0038] Preparation method:

[0039] (1) According to the percentage by weight, potassium penicillin V, clavulanic acid, potassium dihydrogen phosphate, anhydrous magnesium sulfate, disodium EDTA, and anhydrous glucose are sieved separately, and the sieve mesh number is 80 mesh.

[0040] (2) The sieved materials are mixed to finally obtain penicillin V potassium soluble powder.

[0041] Example 3

[0042] A soluble powder for preventing and treating necrotic enteritis in chickens. The soluble powder comprises 50 kg of penicillin V potassium, 9.2 kg of clavulanic acid, 4.6 kg of sodium dihydrogen phosphate, 4.0 kg of talcum powder, 0.1 kg of disodium EDTA, and anhydrous glucose added to 100 kg.

[0043] Preparation method:

[0044] (1) According to the percentage by weight, potassium penicillin V, clavulanic acid, sodium dihydrogen phosphate, talcum powder, disodium EDTA, and anhydrous glucose are sieved separately, and the sieve mesh number is 80 mesh.

[0045] (2) The sieved materials are directly mixed to finally obtain penicillin V potassium soluble powder.

[0046] Example 4

[0047] A soluble powder for preventing and treating necrotic enteritis in chickens. The soluble powder comprises 40 kg of potassium penicillin V, 8.0 kg of sulbactam, 3.0 kg of potassium dihydrogen phosphate, 3.0 kg of anhydrous magnesium sulfate, 0.06 kg of EGTA, and anhydrous glucose added to 100 kg.

[0048] Preparation method:

[0049] (1) According to the percentage by weight, potassium penicillin V, sulbactam, potassium dihydrogen phosphate, anhydrous magnesium sulfate, EGTA, and anhydrous glucose were sieved separately, and the sieve mesh number was 80 meshes.

[0050] (2) The sieved materials are mixed to finally obtain penicillin V potassium soluble powder.

[0051] Example 5

[0052] A soluble powder for preventing and treating necrotic enteritis in chickens. The soluble powder comprises 40 kg of potassium penicillin V, 8.0 kg of tazobactam, 3.0 kg of potassium dihydrogen phosphate, 3.0 kg of anhydrous magnesium sulfate, 0.06 kg of NTA, and anhydrous glucose added to 100 kg.

[0053] Preparation method:

[0054] (1) According to the percentage by weight, penicillin V potassium, tazobactam, potassium dihydrogen phosphate, anhydrous magnesium sulfate, NTA, and anhydrous glucose were sieved separately, and the sieve mesh number was 80 meshes.

[0055] (2) The sieved materials are mixed to finally obtain penicillin V potassium soluble powder.

[0056] Example 6

[0057] A soluble powder for preventing and treating necrotic enteritis in chickens. The soluble powder comprises 40 kg of potassium penicillin V, 8.0 kg of clavulanic acid, 3.0 kg of anhydrous magnesium sulfate, 0.06 kg of penicillamine, and anhydrous glucose added to 100 kg.

[0058] Preparation method:

[0059] (1) According to the percentage by weight, potassium penicillin V, clavulanic acid, anhydrous magnesium sulfate, penicillamine, and anhydrous glucose are sieved separately, and the sieve mesh number is 80 mesh.

[0060] (2) The sieved materials are mixed to finally obtain penicillin V potassium soluble powder.

[0061] Comparative Example 1

[0062] A veterinary penicillin V potassium soluble powder is prepared according to the following steps:

[0063] (1) Crushing and sieving: according to the weight percentage of the components, take 50 g of potassium penicillin V, 30 g of citric acid, 3 g of succinic acid, and 17 g of glucose; crush them respectively with a crusher and sieve them with a mesh size of 120;

[0064] (2) Mixing: First, premix 50 g of crushed penicillin V potassium and 30 g of citric acid, and then mix with 3 g of crushed succinic acid and 17 g of glucose to finally obtain the penicillin V potassium soluble powder.

[0065] Test Example 1 Penicillin V Potassium Stability Test in Water

[0066] 1. Effect of tap water quality on the degradation of penicillin V potassium

[0067] In order to evaluate the causes of the degradation of penicillin V when dissolved in tap water, the effects of tap water of different qualities were tested. Different concentrations of ions were added to samples containing 1 mg / ml penicillin V potassium, the samples were placed at room temperature for 24 hours, and the penicillin V content was measured at 12 hours and 24 hours.

[0068] Table 1 Summary of penicillin V content changes

[0069]

[0070] Although no obvious physical changes were observed in the various solutions, the decreases in content varied, as can be clearly seen from the results, especially in Ca + 、Al 3+ , Fe 3+ It has a relatively large impact on the decrease in penicillin V content, and the impact is greater under alkaline conditions.

[0071] 2. Detection of penicillin V content in water

[0072] Test method: The penicillin V potassium soluble powder prepared in Examples 1-6 and Comparative Example 1 was used to detect the penicillin V content in water according to the penicillin V potassium content detection method in the "Chinese Pharmacopoeia 2020" version (the water used was laboratory tap water). The content change values ​​are shown in the following table:

[0073] Table 2 Summary of penicillin V potassium stability test in water

[0074]

[0075]

[0076] The results show that: Examples 1, 2, and 3 can all achieve a 12-hour aqueous solution stability of more than 99%. The stability is significantly better than that of Comparative Example 1. As can be seen from Example 6, if a pH regulator is not added, the stability will deviate, and in actual use, the water pH in different regions also varies. Adding a pH regulator can make the system more stable. The penicillin V potassium soluble powder used for preventing and treating necrotic enteritis in chickens of the present invention has greatly improved stability under the action of penicillin V potassium and a β-lactam inhibitor, and has good stability, especially Examples 1-3 have better effects, especially Example 2 is the best, and Comparative Example 1 has a significantly poorer effect.

[0077] Test Example 2: Content stability test of water samples from different regions in different embodiments

[0078] Water sample collection: Our marketing personnel collect water samples from different regions without processing.

[0079] Test plan: Examples 1, 2, 3 and Comparative Example 1 were dissolved at the highest clinical concentration, placed at room temperature, and the penicillin V content in the solution was detected at 0h, 12h and 24h. The results are shown in the table below.

[0080] Table 3 Summary of penicillin V content changes in penicillin V potassium soluble powder in different regions / %

[0081]

[0082] The experimental results show that the stability of comparative example 1 is slightly worse in water samples from different regions, and basically does not meet the requirements (the required content is 90-110% qualified) within 24 hours, and decreases rapidly. The penicillin V content of water samples from different regions detected at 0 hours in Examples 1-3 is slightly different, but within the standard range. After 24 hours of placement, it can be seen that the penicillin V content of water samples from different regions can be guaranteed to be above 99% at 12 hours, and the content decreases slightly after 24 hours, but can be guaranteed to be above 95%. The content of penicillin V potassium soluble powder in water of the present invention is relatively stable.

[0083] Test Example 3 In vitro antibacterial activity evaluation test

[0084] Culture medium: Use nutrient broth culture medium, weigh 21g of culture medium, heat and dissolve it in 1000ml of purified water, sterilize it at 121℃ for 20min, and set aside. RCM culture medium, weigh 38g of culture medium, heat and stir and dissolve it in 1000ml of purified water, sterilize it at 121℃ for 15min, and set aside.

[0085] Preparation of bacterial solution: Staphylococcus aureus, Escherichia coli, and Clostridium perfringens are all from clinical isolates. The above strains are revived and typical colonies are selected. Staphylococcus aureus and Escherichia coli are cultured at 37°C for 18-24 hours for later use; Clostridium perfringens is cultured in a culture bag equipped with an anaerobic gas production bag at 37°C for 18-24 hours for later use.

[0086] Preparation of test solution: Weigh a certain amount of antibacterial drug penicillin V potassium and prepare it to a concentration of 512 μg / ml as the first dilution drug for standby use. Subsequent drugs are diluted with this as the mother solution. Prepare 1 portion each of Examples 2, 4, 5 and Comparative Example 1.

[0087] Preparation of drug solution dilution and bacterial solution inoculation: The two-fold dilution method was used for dilution. Each clinical strain was diluted according to the following method. Taking Staphylococcus aureus as an example, 12 sterile test tubes were taken and numbered, and 2.0 ml of broth was added to each tube. 2.0 ml of potassium penicillin V drug solution was added to test tube No. 1 and mixed by pumping. Subsequently, 2.0 ml was transferred to test tube No. 2 and mixed by pumping. Then, 2.0 ml was transferred from test tube No. 2 to test tube No. 3. Continuously diluted to No. 10, and 2.0 ml was transferred from test tube No. 10 and discarded. Test tube No. 11 was not added with drug solution but bacterial solution as a positive control. Test tube No. 12 was not added with drug solution or bacterial solution, but only with culture medium as a negative control. The final drug concentrations of each test tube are 32μg / ml, 16μg / ml, 8μg / ml, 4μg / ml, 2μg / ml, 1μg / ml, 0.5μg / ml, 0.25μg / ml, 0.125μg / ml, and 0.0625μg / ml, respectively. Each tube is parallel to 2. Dilute the standby bacterial solution 100 times with broth, take 0.1ml of bacterial dilution and add it to the above test tubes 1 to 11 respectively and mix evenly, add 0.1ml of sterile saline to test tube 12. Plug the test tubes with inoculation numbers, place them in an incubator and culture for 24 hours, and observe the results.

[0088] Results inspection: The positive control tube No. 11 is turbid, and the negative control tube No. 12 is transparent. On the premise that both meet the requirements, observe the turbidity of other test tubes. If it is turbid, it means that bacteria are growing. If it is transparent, it proves that there is no bacterial growth. The lowest drug concentration contained in the test tube without bacterial growth is the MIC.

[0089] Table 4 Summary of in vitro antibacterial test results (μg / ml)

[0090] strain Example 2 Example 4 Example 5 Comparative Example 1 Standard Clostridium perfringens ≦0.0625 ≦0.0625 ≦0.0625 ≦0.0625 Clinical Clostridium perfringens (n=20) 0.125 2 4 1 Clinical Escherichia coli (n=15) 64 256 256 >512 Clinical Staphylococcus aureus (n=12) ≦0.0625 4 2 32

[0091] The results show that different samples of clinical Clostridium perfringens are all sensitive, and the antibacterial effect is significantly improved after adding β-lactam inhibitors, and Example 2 has a good inhibitory effect on clinical Staphylococcus aureus. Clinical Escherichia coli has relatively high drug resistance, and Comparative Example 1 shows drug resistance. After adding β-lactam inhibitors, the sensitivity is improved to varying degrees, and Example 2 has greatly improved sensitivity compared with other samples.

[0092] The above description is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Any modification, equivalent substitution and improvement made by any technician familiar with the technical field within the technical scope of the present invention and within the spirit and principle of the present invention should be covered within the protection scope of the present invention.

Claims

1. A soluble powder for preventing and treating necrotic enteritis in chickens, characterized in that: The soluble powder consists of penicillin V potassium, beta-lactam inhibitor, pH regulator, anti-caking agent, metal ion chelating agent and filler.

2. The soluble powder according to claim 1, characterized in that The components and their weight percentages in the soluble powder are 30-80% of potassium penicillin V, 1-20% of beta-lactam inhibitor, 0.5-6% of pH regulator, 0-10% of anti-caking agent, 0.02-0.5% of metal ion chelating agent, and the filler is added to 100%.

3. The soluble powder according to claim 2, characterized in that The components and their weight percentages in the soluble powder are 35-50% of potassium penicillin V, 3.5-9.2% of beta-lactam inhibitor, 0.6-4.6% of pH regulator, 2-4% of anti-caking agent, 0.02-0.1% of metal ion chelating agent, and the filler is added to 100%.

4. The soluble powder according to any one of claims 1 to 3, characterized in that: The β-lactam inhibitor is one or more of clavulanic acid, sulbactam, tazobactam, avibactam, ralibactam and vaborbactam.

5. The soluble powder according to claim 4, characterized in that The β-lactam inhibitor is clavulanic acid.

6. The soluble powder according to any one of claims 1 to 3, characterized in that: The pH regulator is one or more of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, and disodium hydrogen phosphate.

7. The soluble powder according to any one of claims 1 to 3, characterized in that: The anti-caking agent is one or more of anhydrous magnesium sulfate, polyvinyl alcohol, sodium lauryl sulfate and talc.

8. The soluble powder according to any one of claims 1 to 3, characterized in that: The filler is one or more of maltodextrin, lactose, anhydrous glucose, β-cyclodextrin, sucrose and fructose.

9. The soluble powder according to any one of claims 1 to 3, characterized in that: The metal ion chelating agent is one or more of disodium EDTA, penicillamine, nitrilotriacetic acid, diisopropylaminetetraacetic acid, and ethylenediaminetetraacetic acid.

10. The soluble powder according to claim 3, characterized in that The components and their weight percentages in the soluble powder are 40% of potassium penicillin V, 8% of clavulanic acid, 3% of pH regulator, 3% of anti-caking agent, 0.06% of disodium EDTA, and anhydrous glucose is added to 100%.

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