CN102697784B - Enrofloxacin injection for livestock and preparation method thereof - Google Patents

Enrofloxacin injection for livestock and preparation method thereof Download PDF

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CN102697784B
CN102697784B CN201210195452.0A CN201210195452A CN102697784B CN 102697784 B CN102697784 B CN 102697784B CN 201210195452 A CN201210195452 A CN 201210195452A CN 102697784 B CN102697784 B CN 102697784B
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enrofloxacin
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CN102697784A (en
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张卫元
张永丹
操继跃
陈翠兰
肖飞
周瑜
刘国庆
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Wuhan Sheng Sheng biological Polytron Technologies Inc
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WUHAN HVSEN BIOTECHNOLOGY CO Ltd
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Abstract

The invention relates to the technical field of a pharmaceutical preparation for livestock and specifically discloses an enrofloxacin injection for the livestock and a preparation method thereof. The raw material of the injection is composed of enrofloxacin, artesunate, trimethoprim, flunixin meglumine, L-arginine, antioxidant, chelant, compound organic solvent and water for injection. The enrofloxacin injection for the livestock can effectively enhance the antibacterial effect of the drug, has an excellent anti-inflammatory effect and is mainly used for treating serious mixed infection caused by sensitive bacteria and mycoplasma of the livestock; the generation of drug resistance is effectively reduced; and the toxic and side effects are reduced. The preparation method for the enrofloxacin injection for the livestock provided by the invention is simple, low-cost, high in stability, convenient in use, and suitable for industrial production.

Description

A kind of enrofloxacin injection for animals and preparation method thereof
Technical field
The present invention relates to veterinary drug preparation technique field, be specifically related to a kind of enrofloxacin injection for animals, relate to the preparation method of this injection simultaneously.
Background technology
Enrofloxacin is the third generation fluoroquinolones extensive pedigree antibiotic of animal specific, has broad-spectrum bactericidal action, and gram negative bacilli and coccus, positive bacteria, mycoplasma, chlamydia are all had to killing action.All effective to the bacterial body of resting stage and trophophase.Sensitive organism is dead in 20-30 minute after contact this product.Bactericidal activity is strong, efficient, and low toxicity is clinically mainly used in treating poultry antibacterial, the caused enteritis of mycoplasma, pneumonia, mastitis and soft tissue infection's property disease.
Meinen etc. (1995) research shows that enrofloxacin bactericidal action has concentration dependent, in tissue and blood plasma, the concentration of enrofloxacin must be higher and maintains the sufficiently long time and could play inhibitory action to target pathogen, enrofloxacin is due to minimal inhibitory concentration MIC relatively low (MIC of most sensitive strains is all less than to 1 μ g/m L), and close with minimal bactericidal concentration MBC, therefore there is very strong antibacterial activity.But be not difficult to find out, improve enrofloxacin concentration, just can increase the bactericidal effect of enrofloxacin.
Along with the heavy dose of this medicine is used and abuses, cause many pathogen to produce drug resistance to this medicine, antibacterial activity is declined, clinical effectiveness is worse and worse.Enrofloxacin concentration is also by initial 0.5%(m/v, g/mL) progressively develop into present 10%(m/v, g/mL), at present again oriented 15%, 20% or the trend of higher concentration development.Because the dissolubility of enrofloxacin in alkaline solution is limited, in production, be prone to the problems such as crystallization, frost resistance be poor, will be larger to the difficulty of high concentration challenge like this.In addition, quinolones is widely distributed in vivo, in order to reach effective therapeutic effect, increases drug dose, easily like this causes that medicine is residual in its hetero-organization to be increased and produce some toxic and side effects.
Summary of the invention
For the deficiencies in the prior art, first object of the present invention is to provide a kind of enrofloxacin injection for animals, this injection is antibacterial, antiphlogistic effects good, be mainly used in treating poultry sensitive bacterial, the caused serious mixed infection of mycoplasma, and can effectively reduce the generation of drug resistance, reduce toxic and side effects.
Second object of the present invention has been to provide a kind of preparation method of above-mentioned enrofloxacin injection for animals, and the product stability that the method makes is high, and cost is low, simple, is applicable to industrialized great production.
The technical scheme that realizes first object of the present invention is as follows:
A kind of enrofloxacin injection for animals, is characterized in that, every this injection of 100mL composed as follows:
Enrofloxacin 2-10g,
Artesunate 0.2-1g,
Trimethoprim 0.4-2g,
Flunixin meglumine 0.1-0.5g,
L-arginine 5-10g,
Antioxidant 0.1-0.3g,
Chelating agen 0.01-0.05g,
Compounded organic solvent 15-60mL,
Surplus is water for injection;
Wherein, the mass ratio of enrofloxacin and artesunate is 10:1, and the mass ratio of enrofloxacin and trimethoprim is 5:1;
Wherein, antioxidant is sodium sulfite or sodium thiosulfate;
Wherein, chelating agen is sodium ethylene diamine tetracetate;
Described compounded organic solvent is the mixture of propylene glycol and ethanol, and both volume ratios are 1:1-2:1.
The technical scheme that realizes second object of the present invention is as follows:
A preparation method for above-mentioned enrofloxacin injection for animals, its step is as follows:
(1) L-arginine is added in water for injection, be heated to 50-60 ℃ of dissolving, after having dissolved, add the enrofloxacin A liquid that stirs to obtain;
(2) antioxidant and chelating agen are under agitation dissolved in water for injection, obtain B liquid;
(3) propylene glycol is heated to 75-85 ℃, adds trimethoprim, after dissolving completely, be cooled to 30 ℃, obtain C liquid;
(4) flunixin meglumine is dissolved completely with water for injection, obtain D liquid;
(5) artesunate is complete with dissolve with ethanol, obtain E liquid;
(6) B liquid is poured in A liquid, fully mix homogeneously, after solution is clear and bright, add C liquid, D liquid and E liquid, surplus water for injection polishing, after medicinal liquid is clear and bright again, if desired, regulating pH value to L-arginine in 9.5-10.5(raw material with ethanolamine is cosolvent and pH adjusting agent, thus ethanolamine only with little amount or need not), filter, filtrate embedding sterilizing, obtains enrofloxacin injection for animals of the present invention.
It is the cosolvent of enrofloxacin injection that the present invention adopts L-arginine, and L-arginine here or pH adjusting agent and solution stabilizer.
In the present invention, added the artesunate with antiinflammatory and antibacterial sensitization, artesunate is the derivant with the antimalarial arteannuin of sesquiterpene structure, chemistry artesunate by name.Artemisine compounds, except being the drug of first choice for the treatment of malaria, also has a lot of valuable pharmacological activity, as resisting pathogenic microbes (schistosomicide, toxoplasma, Eperythrozoon etc.), heat clearing away, antiinflammatory, antiviral and immunoregulation effect etc.The independent use of artesunate does not almost have antibacterial action, but has obvious antibacterial sensitization while using from different Antibiotic combinations, can reduce antibiotic using dosage; Obviously reduce antibacterial (comprising Gram-negative, positive bacteria), colibacillus deactivating attack mouse death rate, significantly suppress the release of mice serum LPS content and proinflammatory cytokine TNF-α, IL-6.Artesunate combined with antibiotic treatment antibacterial infectious effect is better.Have no at present the report of artesunate associating enrofloxacin.On veterinary clinic, artesunate is equally mainly used in the multiple distemper epidemic disease that the rickettsia such as poultry, protozoon, antibacterial, virus etc. cause with arteannuin, and activity is higher.
Select synergistic factor trimethoprim (Trimethoprim, TMP), can greatly strengthen the antibacterial activity of the Multiple Classes of Antibiotics such as sulfonamides, quinolones medicine, its mechanism of action is to suppress dihydrofolate reductase, make dihydrofoilic acid can not be reduced to tetrahydrofolic acid, thereby block responsive folic acid metabolism and utilization, thereby overslaugh thalline nucleic acid is synthetic.Enrofloxacin is to bring into play antibacterial action by the activity blocking-up bacterium DNA replication of anti-bacteria DNA helicase and Topoisomerase Ⅳ.The two associating, can block bacterial growth simultaneously and breed necessary nucleic acid and albumen building-up process, thereby increases antibacterial effect, reduces the appearance of Resistant strain.
Flunixin meglumine is a kind of potent cox-2 inhibitors, has analgesia, antipyretic, antiinflammatory and anti rheumatism action.When poultry are subject to antibacterial, virus, mycoplasma invasion and attack morbidity, clinical conventionally with inflammatory symptoms (heating, pain), thus affect the normal diet of poultry and healthy etc., therefore be necessary antiinflammatory (antipyretic, ease pain) in antibacterial, antiviral process.Flunixin meglumine is independent or can obviously improve clinical symptoms with Antibiotic combination medication, and can strengthen antibiotic activity.On veterinary clinic, be usually used in alleviating pain and antiinflammatory that internal organs angor, muscle and the skeleton disorder of horse cause; The various diseases of cattle infects the acutely inflamed control causing, as laminitis, arthritis etc., also can be used in addition the syndromic auxiliary treatment of sow mastitis, metritis and agalactia.
Compared with prior art, advantage of the present invention and beneficial effect are as follows:
1, the present invention has selected to have artesunate and the coupling of antibiotic enrofloxacin of antiinflammatory and antibacterial sensitization, has strengthened the antibacterial strength of enrofloxacin.
2, adding of Trimethoprim trimethoprim (TMP), make the antibacterial activity of enrofloxacin strengthen 3-6 doubly.
3, flunixin meglumine is analgesia, antipyretic, antiinflammatory and antirheumatic, and untoward reaction is few, and dosage is light, strong drug action, and individual variation is little, remarkable for poultry heating, painful diseases effect.
4, according to each feedstock property, this injection has been selected suitable solvent, antioxidant, chelating agen, strengthened the stability of enrofloxacin injection for animals, and frost resistance is fine.
Enrofloxacin injection wide spectrum for animals of the present invention, efficient, can be used for treating poultry sensitive bacterial, the caused serious mixed infection of mycoplasma, can be widely used in the treatment of poultry infectious disease.
The specific embodiment
Below applicant in connection with specific embodiment, the present invention is described in further detail so that those skilled in the art understands the present invention better, but following content should not be understood to the restriction to the claims in the present invention book request protection domain.
Embodiment 1:2.5m/v%(g/mL) enrofloxacin injection preparation for animals
A kind of enrofloxacin injection for animals, the formula of every this injection of 100mL is as follows:
Enrofloxacin 2.5g,
Artesunate 0.25g,
Trimethoprim 0.5g,
Flunixin meglumine 0.125g,
L-arginine 5g,
Sodium sulfite 0.1g,
Sodium ethylene diamine tetracetate 0.01g,
Propylene glycol 10mL,
Ethanol 5mL,
Water for injection adds to 100mL.
The enrofloxacin injection for animals of above-mentioned formula is prepared in accordance with the following steps and is obtained:
1) L-arginine is added in 20mL water for injection, be heated to 55 ℃ of dissolvings, after having dissolved, add the enrofloxacin A liquid that stirs to obtain.
2) sodium sulfite and sodium ethylene diamine tetracetate are under agitation dissolved in 5mL water for injection, obtain B liquid.
3) propylene glycol is heated to 80 ℃, adds trimethoprim, after dissolving completely, be cooled to 30 ℃, obtain C liquid.
4) flunixin meglumine is dissolved completely with 5mL water for injection, obtain D liquid.
5) artesunate is complete with dissolve with ethanol, obtain E liquid.
6) B liquid is poured in A liquid, fully mix homogeneously, after solution is clear and bright, add again C liquid, D liquid and E liquid, add again the water for injection of surplus to 100mL, after medicinal liquid is clear and bright,, filter to 10.3-10.4 with ethanolamine adjusting pH value, filtrate embedding sterilizing, obtains 2.5m/v%(g/mL) enrofloxacin injection for animals.
Embodiment 2:5m/v%(g/mL) enrofloxacin injection preparation for animals
A kind of enrofloxacin injection for animals, the formula of every this injection of 100mL is as follows:
Enrofloxacin 5g,
Artesunate 0.5g,
Trimethoprim 1g,
Flunixin meglumine 0.25g,
L-arginine 8g,
Sodium thiosulfate 0.2g,
Sodium ethylene diamine tetracetate 0.02g,
Propylene glycol 20mL,
Ethanol 15mL,
Water for injection adds to 100mL.
The preparation process of injection is with embodiment 1.
Embodiment 3:7.5m/v%(g/mL) enrofloxacin injection preparation for animals
A kind of enrofloxacin injection for animals, the formula of every this injection of 100mL is as follows:
Enrofloxacin 7.5g,
Artesunate 0.75g,
Trimethoprim 1.5g,
Flunixin meglumine 0.375g,
L-arginine 9g
Sodium thiosulfate 0.25g,
Sodium ethylene diamine tetracetate 0.03g,
Propylene glycol 25mL,
Ethanol 15mL,
Water for injection adds to 100mL.
The preparation process of injection is with embodiment 1.
Embodiment 4:10m/v%(g/mL) enrofloxacin injection preparation for animals
A kind of enrofloxacin injection for animals, the formula of every this injection of 100mL is as follows:
Enrofloxacin 10g,
Artesunate 1g,
Trimethoprim 2g,
Flunixin meglumine 0.5g,
L-arginine 10g,
Sodium thiosulfate 0.3g,
Sodium ethylene diamine tetracetate 0.05g,
Propylene glycol 30mL,
Ethanol 30mL,
Water for injection adds to 100mL.
The preparation process of injection is with embodiment 1.
Test example 1: antibacterial activity comparative test
Object: relatively whether artesunate can improve the antibacterial activity of enrofloxacin.
Control sample: enrofloxacin crude drug and common 5% enrofloxacin injection, all purchased from Haikang, Guangdong Province animal pharmaceutical factory.
Test specimen: prepare enrofloxacin injection for animals by the method for embodiment 2, but do not add trimethoprim, flunixin meglumine.
Adopt test tube doubling dilution, measure respectively enrofloxacin raw material, common 5% enrofloxacin injection, test specimen to escherichia coli ATCC 25922with staphylococcus aureus ATCC 25923minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC), with the antibacterial effect of comparative test sample and control sample, the results are shown in Table 1.
Table 1 antibacterial activity comparative test result
Figure BDA00001764729200071
As shown in Table 1, the enrofloxacin injection that contains artesunate is to escherichia coli ATCC 25922with staphylococcus aureus ATCC 25923antibacterial activity be control sample crude drug and ordinary preparation 2 times, result shows that artesunate has strengthened the antibacterial activity of enrofloxacin, compared with its ordinary preparation, the enrofloxacin injection of having added artesunate has stronger antibacterial action.
Test example 2: the stability test of enrofloxacin injection for animals of the present invention
Method: get respectively enrofloxacin injection product for animals prepared by embodiment 1-4, carry out accelerated test according to appendix veterinary drug stability test guideline of Chinese veterinary pharmacopoeia version in 2010, press commercially available back, under the condition of 40 ± 2 ℃ of temperature, relative humidity 75 ± 5%, place 6 months, sample respectively once the 1st, 2,3,6 the end of month at duration of test, and appearance character, pH value and content (by the percentage composition of labelled amount) to injection detect.
Stability test result: after the accelerated test of 6 months, all kinds of indexs of product, refer to following table.
Embodiment 1 product
Figure BDA00001764729200072
Embodiment 2 products
Figure BDA00001764729200073
Figure BDA00001764729200081
Embodiment 3 products
Figure BDA00001764729200082
Embodiment 4 products
Figure BDA00001764729200083
As can be seen from the above data, after the product accelerated test of 6 months, character, the pH value of product of the present invention have no significant change, determining enrofloxacin content also meets the regulation within the scope of the 95%-105% that national veterinary drug standard active ingredient is labelled amount, illustrates that this product is very stable under this experimental condition.
Test example 3: the antifreezing test of enrofloxacin injection for animals of the present invention
Method: get enrofloxacin injection product for animals prepared by embodiment 1-4, press commercially available back, be placed on respectively 4 ± 2 ℃ of cold preservations freezing with-20 ± 2 ℃, sample respectively once the 1st, 2,3,6 the end of month in experimental period, and the appearance character to injection, the situation of thawing (room temperature is thawed) and content (by labelled amount percentage composition) detect.
Antifreezing test result: after the accelerated test of 6 months, all kinds of indexs of product, refer to following table.
Embodiment 1 product
Figure BDA00001764729200091
Embodiment 2 products
Figure BDA00001764729200092
Embodiment 3 products
Figure BDA00001764729200093
Embodiment 4 products
Figure BDA00001764729200101
As can be seen from the above data, cold preservation under cryogenic conditions, product characteristics are good, and without separating out, determining enrofloxacin content meets national veterinary drug standard (95%-105% that effective ingredient is labelled amount) regulation; And-20 ± 2 ℃ freezing 6 months, first 2 months are stable, although having occurred crystallization more or less the 3-6 month, all can redissolve, assay also meets the requirements, and the low temperature resistant of product is described, frost resistance is respond well.
Test example 4: clinical trial
Use the product of embodiment 2 to test, the 5% enrofloxacin injection group that matched group selects Haikang, Guangdong Province animal pharmaceutical factory to produce, enrofloxacin consumption: pig: intramuscular injection 10-20mg(enrofloxacin meter), 2 times/day, successive administration 2-3 days.Chicken: 150-200mL converts 100kg water, freely drinks 1-2 times/day, successive administration 2-3 days for chicken.Cattle: intramuscular injection 5-10mg(enrofloxacin meter), 1 times/day, successive administration 2-3 days; Mammitis of cattle adopts breast perfusion, each newborn chamber 250mg(enrofloxacin meter), 1 times/day, successive administration 2-3 days.Test data is as follows:
The clinical therapeutic efficacy of embodiment 2 products
Figure BDA00001764729200102
Clinical therapeutic efficacy comparative result
Figure BDA00001764729200111
Test example 5: clinical trial
500 of pigs are raised on certain pig farm, in the time of July, fall ill, and show as fervescence, the decline of gradual appetite, vomiting, and the sleeping ground of pig does not rise, and shivers, walk haltingly on foot, some arthroncuss, some appearance nervous symptoms, some mouthful is flowed foam sample material.Eyes circle is sent out crow, and conjunctivitis is serious, and some eye conjunctivas are pale or rubescent.Cough, rhinorrhea, significantly breathe, serious dyspnea (the similar shape sound of snoring), doubt as the high heat of the mixed sense of pig (being caused by pig circular ring virus, reproductive and respiratory syndrome poison, influenza virus, streptococcus and the mixed infection of secondary haemophilus intersection), the enrofloxacin injection for animals that adopts embodiment 4 to prepare, intramuscular injection, by per kilogram of body weight 15-20mg(enrofloxacin), every day 2 times, successive administration 3-4 days, cure rate 95.62%.
Safety testing result: after injection, sick pig, without abnormal dead, is observed 30 days, the trouble poultry after administration grows fine, without any sick generation.
Show that by above experiment this product has obvious therapeutic effect to infection such as treatment poultry sensitive bacterial, the caused digestive system of mycoplasma, respiratory system, urinary system infection and pig toxoplasmosis, Eperythrozoons, suffers from poultry to inject rear rapid-action; Treat mixed infection and the severe infections that multiple pathogenic bacteria, mycoplasma cause and have good therapeutic effect.
Safety experiment shows, product of the present invention is showed no toxic and side effects.

Claims (1)

1. an enrofloxacin injection for animals, is characterized in that, the preparation method step of described injection is as follows:
(1) 5-10g L-arginine is added in water for injection, be heated to 50-60 ℃ of dissolving, after having dissolved, add the 2-10g enrofloxacin A liquid that stirs to obtain;
(2) 0.1-0.3g antioxidant and 0.01-0.05g sodium ethylene diamine tetracetate are under agitation dissolved in water for injection, obtain B liquid;
(3) propylene glycol is heated to 75-85 ℃, adds 0.4-2g trimethoprim, after dissolving completely, be cooled to 30 ℃, obtain C liquid;
(4) 0.1-0.5g flunixin meglumine is dissolved completely with water for injection, obtain D liquid;
(5) 0.2-1g artesunate is complete with dissolve with ethanol, obtain E liquid;
(6) B liquid is poured in A liquid, fully mix homogeneously, adds C liquid, D liquid and E liquid again, then adds water for injection to 100mL after solution is clear and bright, after medicinal liquid is clear and bright, if desired, with ethanolamine regulate pH value to 9.5-10.5, filter, filtrate embedding sterilizing, obtains described enrofloxacin injection for animals;
The mass ratio of described enrofloxacin and artesunate is 10:1, and the mass ratio of enrofloxacin and trimethoprim is 5:1;
Described antioxidant is sodium sulfite or sodium thiosulfate;
Described propylene glycol and ethanol volume ratio are 1:1-2:1, described propylene glycol and ethanol composition compounded organic solvent, and compounded organic solvent volume is 15-60mL.
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CN103126982A (en) * 2013-03-07 2013-06-05 庞永中 Novel veterinary medicament meglumine enrofloxacin injection and preparation method thereof
CN103222978B (en) * 2013-04-02 2018-03-06 河南省乘风动物保健技术研究所 Fu side's Sulfamethoxazole parenteral solution and preparation method
CN104398586A (en) * 2014-12-18 2015-03-11 郑州福源动物药业有限公司 Shuanghuanglian oral liquid for livestock and poultry as well as preparation method of Shuanghuanglian oral liquid
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CN105267142B (en) * 2015-10-15 2018-02-09 四川鼎尖动物药业有限责任公司 A kind of enrofloxacin injection and preparation method thereof
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