Compound samidazuril preparation and preparation method thereof

Abstract

The invention discloses a compound samipril preparation and a preparation method thereof, and belongs to the technical field of veterinary chemical preparations. The preparation is composed of samizuril, an iron (III) compound, a solvent, a cosolvent, a pH regulator and a bacteriostatic agent. The compound samizuril preparation has good stability and safety and is convenient to take. In the iron supplementing process of the piglets, the insect-resistant drug samizuril is added to effectively kill coccidiosis, the defect that an iron supplementing agent and an insect repellent can only be used independently is overcome, adverse reactions caused by iron supplementing agent injection are reduced, and the iron supplementing agent is good in bioavailability, outstanding in clinical effect and wide in application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of veterinary chemical preparations, and in particular relates to a veterinary compound samizulil preparation and a preparation method thereof. Background Art

[0002] Iron is an important trace element in the growth process of piglets. Hemoglobin needs sufficient iron to be synthesized. If the iron reserves in the piglets are insufficient, they may suffer from anemia, slow growth and other symptoms, which will cause certain economic losses to farmers. Therefore, iron supplementation has become an important measure in the growth process of piglets. In the current farms, in order to prevent and treat iron deficiency in piglets, iron supplements are usually given by intramuscular injection at 1 to 7 days of age. However, the existing iron supplement injections are highly irritating and are prone to adverse reactions such as swelling and redness at the injection site, and may even cause stress in piglets.

[0003] Another disease in the growth process of piglets is coccidiosis, which is caused by Eimeria or Isospora parasitic in the host's intestinal or bile duct epithelial cells. It is common in piglets aged 7-10 days. The main symptoms are diarrhea, fever, loss of appetite, weight loss, emaciation, etc., and in extreme cases, there is death. Although the mortality rate of the disease is low, it is easy to induce intestinal infections caused by other bacteria or viruses. Even after recovery, it may lead to a slowdown in growth and development, which is more harmful to intensive breeding and easily causes large economic losses to the livestock industry.

[0004] After investigation, it was found that the existing farms first injected iron supplements within 1 to 7 days after piglets were born, and then took orally or injected anticoccidial drugs after 3 days of age. Since piglets have two needs for iron supplementation and coccidial removal for a period of time after birth, there is no compound preparation that can meet both needs in the current market, so only anthelmintic drugs and iron supplements can be used separately.

[0005] Currently, iron supplement injections are highly irritating, and reports of adverse reactions of varying degrees are common. Samizurin is a new triazine anticoccidial drug independently developed by the veterinary anti-infective drug innovation team of the Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The drug has shown good anticoccidial effects in previous studies, with low toxicity and high safety. It has no cross-durability with diclazuril and toltrazuril, and has good application development prospects. However, the substance is poorly soluble in water, and its low solubility leads to a low effective concentration of the drug.

[0006] Therefore, piglet farming requires a compound preparation that not only does not produce adverse reactions during iron supplementation, but also has good solubility of anticoccidial active ingredients and can effectively prevent piglet coccidiosis. Summary of the invention

[0007] In order to solve the above technical problems, the present invention provides a compound samizuril preparation and a preparation method thereof, which has good stability, safety and convenient administration. In the process of iron supplementation for piglets, the addition of anthelmintics has a killing effect on coccidia, solves the problem of the poor solubility of the anthelmintic samizuril, and the defect that the iron supplement and the anthelmintic can only be used separately, reduces the adverse reactions caused by the injection of the iron supplement, and overcomes the shortcomings of the prior art.

[0008] The technical solution of the present invention is as follows:

[0009] In one aspect, the present invention provides a compound samizuril preparation, which is composed of the following components: samizuril, an iron (III) compound, a solvent, a cosolvent, a pH regulator, an antibacterial agent, and water for injection.

[0010] Preferably, the dosage of each component in each 100 ml of the compound samizuril preparation is:

[0011] Element Dosage Samizurin 1.0～5.0g Iron(III) compounds 5~20g (based on effective iron) Solvents 20～65ml Solvent 5～30ml pH Adjusters 2.5～10g Antibacterial Agents 0.1～2g Water for injection Refill to 100ml

[0012] More preferably, the dosage of each component in every 100 ml of the compound samizuril preparation is:

[0013] Element Dosage Samizurin 1.5～5.0g Iron(III) compounds 7.5~15g (based on effective iron) Solvents 30～65ml Solvent 10～25ml pH Adjusters 2.5～7.5g Antibacterial Agents 0.1～2g Water for injection Refill to 100ml

[0014] More preferably, the dosage of each component in each 100 ml of the compound samizulil preparation is:

[0015] Element Dosage Samizurin 3.0g Iron(III) compounds 10g (based on effective iron) Solvents 50ml Solvent 10ml pH Adjusters 5g Antibacterial Agents 0.5g Water for injection Refill to 100ml

[0016] Preferably, the iron (III) compound is any one of dextran iron, glucoheptane iron, and sucrose iron.

[0017] More preferably, the iron (III) compound is iron dextran.

[0018] Preferably, the solvent is one or more of water, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, ethyl oleate, and triolein.

[0019] More preferably, the solvent is selected from N,N-dimethylformamide, triolein, dimethyl sulfoxide, ethyl oleate.

[0020] Preferably, the co-solvent is one or more of ethanol, isopropanol, ethyl lactate, propyl lactate, glycerol, and diethylene glycol monoethyl ether.

[0021] More preferably, the co-solvent is selected from ethyl lactate, glycerol, and diethylene glycol monoethyl ether.

[0022] Preferably, the pH adjuster is one or more of phosphate buffer, citric acid, sodium citrate, acetic acid, sodium acetate, tartaric acid, lactic acid, and ethanolamine.

[0023] More preferably, the pH adjuster is selected from phosphate buffer, citric acid, sodium citrate, tartaric acid.

[0024] Preferably, the antibacterial agent is any one of phenol, sodium benzoate, benzyl alcohol and potassium sorbate.

[0025] More preferably, the antibacterial agent is selected from sodium benzoate and potassium sorbate.

[0026] Preferably, the compound samizulil preparation is a compound samizulil injection.

[0027] In another aspect, the present invention also provides a method for preparing a compound samizuril preparation, and the preparation steps are as follows:

[0028] (1) stirring and mixing the solvent and the co-solvent, and adjusting the pH value with an appropriate amount of a pH adjuster to obtain a liquid mixture I;

[0029] (2) dissolving the iron (III) compound and the antibacterial agent in an appropriate amount of water for injection, and adding a pH regulator to adjust the pH value to obtain a liquid mixture II;

[0030] (3) cooling the liquid mixture II to room temperature, mixing it evenly with the liquid mixture I, adding samizurin and dissolving it;

[0031] (4) Make up to volume with water for injection, mix well, divide into packages, and sterilize.

[0032] Preferably, the pH value in step (1) is 8.6±0.5; and the pH value in step (2) is 1.8±0.5.

[0033] Preferably, the temperature of the water for injection in step (2) is 30-60°C, more preferably 50°C.

[0034] The auxiliary materials used in the present invention are all commercially available products.

[0035] The present invention obtains that the age of onset of piglet coccidiosis and neonatal anemia is similar by statistics, and the two drugs are compounded, and solvents such as N, N-dimethylformamide, triolein and cosolvents such as ethanol, glycerol, and diethylene glycol monoethyl ether are added to increase the solubility of samizuli and improve the stability of the preparation; according to the characteristics that the acidic environment can accelerate the hydrolysis of polysaccharides into small molecules, pH regulators such as citric acid are added to the preparation, so that the macromolecular polysaccharides formed after the intramuscular injection of polysaccharide iron decomposition can be hydrolyzed into small molecules in vivo more quickly, thereby reducing the irritation of the injection site. In the final preparation process, the stability of the compounded drug is studied emphatically, and the best ratio for the stability of the efficacy component is obtained.

[0036] Beneficial effects:

[0037] 1. The present invention prepares samizurin into a compound injection, adds a preferred solvent and a co-solvent to improve the solubility of the main component, and the preparation has good long-term storage stability. In addition, the preparation process is relatively simple, the raw and auxiliary materials are relatively common, and the requirements for production equipment are relatively low.

[0038] 2. In the pharmacokinetic test, the preparation of the present invention has a shorter time to peak action, and its maximum blood drug concentration and bioavailability are higher than those of commercially available single-ingredient drug combinations.

[0039] 3. In the irritation test of clinical doses, the compound samizuli injection prepared by the present invention has significantly improved irritation compared with the single-prescription preparations of samizuli injection and polysaccharide iron complex. In actual use, compared with single-prescription preparations, it saves the workload of one-time administration, which is more convenient and quick. The application scenario is not only suitable for the prevention and treatment needs of small-scale farmers, but also can meet the prevention and treatment needs of larger-scale farms.

[0040] 4. The compound samizuril injection prepared by the present invention can prevent and kill coccidia while the iron supplement can effectively improve the immunity of piglets, thereby preventing and treating piglet coccidiosis from two aspects and shortening the treatment cycle. DETAILED DESCRIPTION

[0041] Example 1 A compound samizurin injection and its preparation method

[0042] The formula and dosage of compound samizuli injection:

[0043] Formula Single dose (100ml / bottle) Samizurin 3g Iron Dextran 10g (based on effective iron) N,N-Dimethylformamide 30ml Triolein 20ml Ethyl lactate 10ml Citric acid 4g Sodium Citrate 1g Sodium Benzoate 0.5g Water for injection Make up to 100ml total 100ml

[0044] Preparation method of compound samizuli injection:

[0045] Under stirring conditions, triolein and ethyl lactate were added to N,N-dimethylformamide and the mixture was dissolved until it was clear, and appropriate amounts of citric acid and sodium citrate were added to adjust the pH to 8.6±0.5 to obtain liquid mixture I. Under stirring conditions, water for injection was heated to 50°C, and iron dextran and sodium benzoate were added to dissolve until it was clear, and citric acid and sodium citrate were used to adjust the pH to 1.8±0.5 to obtain liquid mixture II. After the liquid mixture II was cooled to room temperature, liquid mixture I was added and mixed evenly, samizurin was added, and after dissolving and clarifying, the volume was made up with water for injection, and after the volume was made up, stirring was continued until uniform, and then the liquid mixture was packaged and sterilized to obtain the product.

[0046] Example 2 A compound samizurin injection and its preparation method

[0047] The formula and dosage of compound samizuli injection:

[0048] Element Single dose (100ml / bottle) Samizurin 3g Iron Dextran 10g (based on effective iron) Dimethyl sulfoxide 50ml glycerin 10ml Phosphate buffer 5g Sodium Benzoate 0.5g Water for injection Make up to 100ml total 100ml

[0049] Preparation method of compound samizuli injection:

[0050] Under stirring conditions, glycerol is added to dimethyl sulfoxide, and an appropriate amount of phosphate buffer is used to adjust the pH to 8.6±0.5 to obtain liquid mixture I; under stirring conditions, the water for injection is heated to 50°C, and then iron dextran and sodium benzoate are added and dissolved until clear, and the pH is adjusted to 1.8±0.5 with a phosphate buffer to obtain liquid mixture II; after the liquid mixture II is cooled to room temperature, the liquid mixture I is added and mixed evenly, samizurin is added, and after dissolving and clarifying, the volume is made up with water for injection, and after the volume is made up, stirring is continued until uniform, and then the liquid mixture is packaged and sterilized to obtain the product.

[0051] Example 3 A compound samizurin injection and its preparation method

[0052] The formula and dosage of compound samizuli injection:

[0053]

[0054]

[0055] Preparation method of compound samizuli injection:

[0056] Under stirring conditions, add ethyl oleate and ethyl lactate to N,N-dimethylacetamide, and use appropriate amount of tartaric acid to adjust the pH to 8.6±0.5 to obtain liquid mixture I; under stirring conditions, heat the water for injection to 50°C, add dextran iron and potassium sorbate to dissolve until clear, and use tartaric acid to adjust the pH to 1.8±0.5 to obtain liquid mixture II; after cooling liquid mixture II to room temperature, add liquid mixture I and mix evenly, add samizurin, dissolve and clarify, and then use water for injection to make up to volume. After making up to volume, continue stirring until uniform, then package and sterilize to obtain.

[0057] Example 4 A compound samizurin injection and its preparation method

[0058] The formula and dosage of compound samizuli injection:

[0059] Element Single dose (100ml / bottle) Samizurin 3g Iron Dextran 10g (based on effective iron) N,N-Dimethylformamide 30ml Triolein 20ml Diethylene glycol monoethyl ether 10ml Citric acid 4g Sodium Citrate 1g Sodium Benzoate 0.5g Water for injection Make up to 100ml total 100ml

[0060] Preparation method of compound samizuli injection:

[0061] Under stirring conditions, triolein and diethylene glycol monoethyl ether were added to N,N-dimethylformamide and the mixture was dissolved until it was clear. Appropriate amounts of citric acid and sodium citrate were used to adjust the pH to 8.6±0.5 to obtain liquid mixture I. Under stirring conditions, water for injection was heated to 50°C, and iron dextran and sodium benzoate were added and dissolved until it was clear. Citric acid and sodium citrate were added to adjust the pH to 1.8±0.5 to obtain liquid mixture II. Liquid mixture II was cooled to room temperature, and then liquid mixture I was added and mixed evenly. Samizurin was added, and after dissolving and clarifying, the mixture was made up to volume with water for injection. After making up to volume, stirring was continued until uniform, and then the mixture was divided and sterilized to obtain the mixture.

[0062] Example 5 A compound samizurin injection and its preparation method

[0063] The formula and dosage of compound samizuli injection:

[0064] Element Single dose (100ml / bottle) Samizurin 5g Iron glucoheptanoate 7.5g (based on effective iron) N,N-Dimethylformamide 40ml Triolein 25ml Ethyl lactate 25ml Citric acid 2g Sodium Citrate 0.5g Sodium Benzoate 2g Water for injection Make up to 100ml total 100ml

[0065] Preparation method of compound samizuli injection:

[0066] Under stirring conditions, add triolein and ethyl lactate to N,N-dimethylformamide and dissolve the mixture until it is clear. Use appropriate amounts of citric acid and sodium citrate to adjust the pH to 8.6±0.5 to obtain liquid mixture I. Under stirring conditions, heat the water for injection to 50°C, add iron glucoheptose and sodium benzoate and dissolve them until they are clear. Use citric acid and sodium citrate to adjust the pH to 1.8±0.5 to obtain liquid mixture II. Cool the liquid mixture II to room temperature and add it to the liquid mixture I to mix evenly. Add samizurin, dissolve and clarify, and then make up to volume with water for injection. After making up to volume, continue stirring until uniform, then package and sterilize to obtain the mixture.

[0067] Example 6 A compound samizurin injection and its preparation method

[0068] The formula and dosage of compound samizuli injection:

[0069]

[0070]

[0071] Preparation method of compound samizuli injection:

[0072] Under stirring conditions, add triolein and ethyl lactate to N,N-dimethylformamide and dissolve the mixture until it is clear. Use appropriate amounts of citric acid and sodium citrate to adjust the pH to 8.6±0.5 to obtain liquid mixture I. Under stirring conditions, heat the water for injection to 50°C, add sucrose iron and sodium benzoate and dissolve them until it is clear. Use citric acid and sodium citrate to adjust the pH to 1.8±0.5 to obtain liquid mixture II. Cool the liquid mixture II to room temperature, add the liquid mixture I and mix evenly, add samizurin, dissolve and clarify, and then make up to volume with water for injection. After making up to volume, continue stirring until uniform, then package and sterilize to obtain the mixture.

[0073] Example 7 Preparation Stability Study

[0074] Examples 1 to 6 which had been sterilized in 100 ml glass bottles were placed under accelerated conditions (40°C ± 2°C, RH 75% ± 5%) for 1, 2, 3, and 6 months to observe changes in the properties and content of the preparations. The test results are shown in Table 1 below:

[0075] Table 1 Preparation stability test results

[0076]

[0077]

[0078] The experiment shows that the contents in Examples 1 to 6 all change to a certain extent, among which the content change in Example 1 is smaller than that in the other examples, and that in Examples 2 and 3 are relatively poor, indicating that the combined use of solvents, co-solvents, pH regulators, etc. in Example 1 can effectively improve the stability of the compound samizulil preparation.

[0079] Example 8 Pharmacokinetic Test

[0080] Pharmacokinetic tests were conducted on the existing single-prescription samizuli injection and Example 1 and Example 2.

[0081] Experimental animals: A total of 9 healthy piglets, weighing 1-1.5 kg, were divided into a control group and 2 experimental groups, with 3 piglets in each group.

[0082] The test period was 7 days. The drug was administered on the 0th day of the test. The control group was injected with single-prescription samizuril injection at a dosage of 20 mg / kgbw; the two test groups were injected with compound samizuril injections of Example 1 and Example 2, respectively, at a dosage of 20 mg / kgbw. Blood was collected from piglets at multiple time points after administration to detect drug concentration.

[0083] Table 2 Pharmacokinetic test results

[0084]

[0085] The above results show that compared with the single-ingredient samizuli injection: Example 1 injection can shorten the peak time, and the maximum blood drug concentration and bioavailability are higher than the single-ingredient samizuli injection. In addition, the maximum blood drug concentration and bioavailability of the injection of Example 1 are also more prominent than those of Example 2.

[0086] Example 9 Irritation Test

[0087] According to the prescription, the single-ingredient preparations in the prescription were selected to compare the irritation and adverse reactions at the injection site. The test method is as follows:

[0088] Experimental animals: 20 healthy piglets were divided into 4 experimental groups: A, B, C, and D.

[0089] The test period was 7 days. Group A served as the control group, and each piglet was injected with normal saline of the average injection volume of Group B; each piglet in Group B was injected with a compound samizuril preparation (Example 1) at 20 mg / kgbw calculated as samizuril; each piglet in Group C was injected with a 10% dextran iron solution of the average injection volume of Group B; each piglet in Group D was injected with 3% samizuril injection at 20 mg / kgbw calculated as samizuril, and the injection sites were scored and recorded on the day of injection and 1, 2, 3, and 7 days after injection.

[0090] Table 3 Test scoring criteria

[0091] Stimulus score No erythema, no edema 0 Barely visible erythema and edema 1 Clear red spots and obvious skin bulges 2 Moderate or severe erythema with skin elevation greater than 1 mm 3

[0092] The test results are as follows:

[0093] Table 4 Irritation test scoring results

[0094] Experimental Group A B C D Day 0 0 0 1 0 Day 1 0 1 2 1 Day 2 0 1 1 1 Day 3 0 0 1 1 Day 7 0 0 0 0

[0095] The experimental results showed that the injection irritation of test group B (i.e., compound samizulil group) was significantly lower than that of test groups C and D.

[0096] Example 10 Clinical effect test

[0097] According to the indications, a test site with suitable climate is selected for clinical comparison. The test method is as follows:

[0098] Experimental animals: A total of 60 healthy piglets were divided into 6 experimental groups: A, B, C, D, E, and F.

[0099] The experimental period was 8 days. On day 0, piglets B, C, D, E, and F were artificially challenged with coccidia. On day 1, the experimental group was treated with medication according to the clinical usage and dosage of each preparation. The diarrhea situation was statistically analyzed in the subsequent 7 days. The feces of each group of pigs within 7 days after medication were collected and the number of coccidia eggs in the feces were recorded according to the saturated saline flotation method.

[0100] Type of medication:

[0101] Experiment Group A: No artificial attack

[0102] Experimental Group B - Treatment with Compound Samizuli Iron Dextran Injection in Example 1

[0103] Trial Group C: Treatment with samizuli injection

[0104] Group D: Iron dextran injection

[0105] Experimental Group E: Treatment with Toltrazuril Suspension + Iron Dextran Solution

[0106] Experimental Group F - Untreated group

[0107] The test results are as follows:

[0108] Table 5 OPG values ​​of coccidia feces in piglets of each group

[0109]

[0110] Table 6 Proportion of diarrhea piglets in the group

[0111]

[0112]

[0113] The results showed that the therapeutic effect of test group B (compound samizul preparation group) was significantly better than that of test group D (only injection of iron supplement preparation), with a significant decrease in OPG value; compared with test group C (only injection of single anticoccidial drug) and test group E (commonly used piglet coccidiosis treatment plan on the market), there was a certain advantage in the speed of OPG value decrease and the number of piglet diarrhea occurrences.

Claims

1. A compound samizuril preparation, characterized in that: The compound samizuril preparation consists of the following components: samizuril, an iron (III) compound, a solvent, a cosolvent, a pH regulator, an antibacterial agent, and water for injection.

2. A compound samuzuril preparation according to claim 1, characterized in that: The dosage of each component in each 100ml of compound samizuli preparation is:

3. A compound samuzuril preparation according to claim 2, characterized in that: The dosage of each component in each 100ml of compound samizuli preparation is:

4. A compound samuzuril preparation according to claim 3, characterized in that: The dosage of each component in each 100ml of compound samizuli preparation is:

5. A compound samuzuril preparation according to any one of claims 1 to 4, characterized in that: The iron (III) compound is any one of dextran iron, glucoheptane iron and sucrose iron.

6. A compound samuzuril preparation according to any one of claims 1 to 4, characterized in that: The solvent is one or more of water, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, ethyl oleate, and triolein.

7. A compound samuzuril preparation according to any one of claims 1 to 4, characterized in that: The co-solvent is one or more of ethanol, isopropanol, ethyl lactate, propyl lactate, glycerol, and diethylene glycol monoethyl ether.

8. A compound samuzuril preparation according to any one of claims 1 to 4, characterized in that: The pH regulator is one or more of phosphate buffer, citric acid, sodium citrate, acetic acid, sodium acetate, tartaric acid, lactic acid, and ethanolamine.

9. A compound samuzuril preparation according to any one of claims 1 to 4, characterized in that: The antibacterial agent is any one of phenol, sodium benzoate, benzyl alcohol and potassium sorbate.

10. A method for preparing a compound samuzuril preparation according to any one of claims 1 to 4, comprising the following steps: (1) firstly, the solvent and the co-solvent are stirred and mixed, and the pH is adjusted with an appropriate amount of a pH adjuster to obtain a liquid mixture I; (2) dissolving the iron (III) compound and the antibacterial agent in an appropriate amount of water for injection, and adding a pH adjuster to adjust the pH to obtain a liquid mixture II; (3) cooling the liquid mixture II to room temperature, mixing it evenly with the liquid mixture I, adding samizurin and dissolving it; (4) Use water for injection to make up to volume, mix evenly, divide into packages, and sterilize.

Images