Background technology
Chicken coccidiosis is the parasitic disease of being produced by the caused serious harm aviculture of Eimeria (Eimeria).The Eimeria that parasitizes in the Intestinum Gallus domesticus road has 7 kinds, is respectively Eimeria tenella (E.tenella), heap type Eimeria (E.acervulina), murder by poisoning Eimeria (E.necatrix), Eimeria maxima (E.maxima), gentle Eimeria (E.mitis), Bu Shi Eimeria (E.brinetti) and precocious Eimeria (E.praecox).Various coccidiosiss are in the entozoic position of chicken digestive tract difference, Eimeria tenella parasitizes caecum, heap type Eimeria parasitizes duodenum, and Eimeria maxima and murder by poisoning Eimeria parasitize small intestinal, and the Bu Shi Eimeria parasitizes small intestinal hypomere and rectum.
Chicken coccidiosis is that a kind of whole world generally takes place the very serious parasitic disease of poultry husbandry harm.Its economic loss that causes is surprising.In the middle of many mass-sending diseases, the sickness rate of chicken coccidiosis is the highest, accounts for 17~20%.Chicken coccidiosis is widely distributed, we can say that just there is chicken coccidiosis in the place of chicken, at intensification chicken farm its sickness rate up to 50~70%; Mortality rate reaches 20~30%, more can reach 80% when serious.
Deccox (decoquinate) is a kind of quinolines class chemistry anticoccidial drug, and its chemical name is 6-oxygen in last of the ten Heavenly stems base-7-ethyoxyl-4-hydroxyquinoline-3-carboxylic acid, ethyl ester.At present relevant document and patent of synthesizing deccox is less, has described to be raw material to nitro guaioxide potassium or to Nitrocatechol sodium etc. as US3485845, by reactions such as hydrogenation, reduction, pyrocondensations, has finally synthesized deccox.Deccox plays a role at the asexual reproductive phase of coccidiosis, and the deccox after the administration stops its growth by disturbing DNA to synthesize after entering the zygoblast cell.Deccox absorbs fast, can reach the valid density of anticoccidial in 1 hour, the concentration level that peaks after 3 days, because of its coccidiosis life cycle promptly begin in early days play a role, thereby effectively avoided the poultry intestinal to suffer damage.In addition deccox also has wide, the anti-ball index advantages of higher of anti-ball spectrum.The coccidiosis that deccox causes for tender, the murder by poisoning of chicken, heap type, huge, displacement and Bu Shi Eimeria etc. clinically has the better prevention effect.
The deccox structural formula
(molecular formula: C
24H
35NO
5Molecular weight: 417.55)
Though deccox has the effect of good wide spectrum anticoccidial, but owing to exist deccox to be insoluble in the shortcoming of water, therefore, the preparation of the relevant deccox of clinical practice both at home and abroad has only a kind of dosage form of deccox pre-mixing agent, the deccox pre-mixing agent can only adopt the spice administration, and can't be by the drinking-water administration.But in home poultry raising, poultry-farm or specialist often adopt the mode of drinking-water administration to carry out fowl diseases prevention and treatment, because the drinking-water administration is easy, time saving and energy saving, can reduce or avoid the chicken stress simultaneously.The clinical practice of deccox has been subjected to greatly restriction like this, therefore inventing a kind of these problems of deccox preparation that can drink water just can be readily solved, the prescription and the technology of deccox solution have been described as CN101129371A, cosolvent 80-99% uses one or more organic solvents such as ethanol, N-Methyl pyrrolidone, dimethyl formamide, and will be heated under 60-90 ℃ the condition and could dissolve, will there be following problem or deficiency like this:
1, cost of supplementary product height: consumption of organic solvent is very big, and the aqueous solution cost is very high relatively.
2, manufacturing expense height: manufacture process needs heating, cooling, and consumes energy is many, the manufacturing cost height.
3, medicine stability is poor, and general deccox will add in the organic solvent under 60-90 ℃ temperature and dissolves, the less stable of deccox under the condition of high temperature, and quality is very difficult controlled.
4, drug safety is poor, and the solvent of deccox solution 80-99% is an organic solvent, and a large amount of organic solvents can cause individual untoward reaction such as allergy.
Summary of the invention
In order to overcome the above problems, the invention provides a kind of deccox suspension formulations that can effectively be dispersed in the water, the deccox suspension is compared with deccox solution with the deccox pre-mixing agent and is had following advantage:
1, the deccox suspension is easy to use, but both spice uses, and also can evenly spread in the water, does not sink in two hours and by the drinking-water administration, also can reach the effect of treatment immediately under chicken is mined massively the situation that appetite reduces greatly by drinking-water.
2, deccox suspension safety of medicine, toxicity is low.Because the solvent of deccox solution 100% is an organic solvent, a large amount of organic solvents can cause individual anaphylaxis etc., and the deccox suspensoid drops to the addition of organic solvent below 20%.
3, deccox suspension cost reduces, and compares with deccox solution, because adopt water to make solvent, has reduced the consumption of organic solvent, so cost reduces greatly.
4, deccox suspension medicine stability height, if with deccox be dissolved in make deccox solution in the solvent after, medicine is oxidation deterioration very easily, but when being to use suspending agent to be dispersed in the solid drugs microgranule in the solvent uniformly, medicine is stable to be improved greatly.
5, dissolution rate is fast than the deccox pre-mixing agent in the deccox suspendible liquid, carrier components such as sodium laurylsulfate, citrate have been used in the deccox suspension preparation process, and be prepared by high pressure homogenizer, obtain the fine particles of different-grain diameter between 1nm~100um, in vivo stripping onset rapidly.
6, deccox suspension medicine drug loading height can increase to 30% with the content of principal agent.
7, deccox suspension good dispersion degree, good stability in water.
Deccox suspension formulations of the present invention is made by the composition of following proportioning:
Deccox 3~30% (W/V)
Wetting agent 1~15% (V/V)
Surfactant 0.01%~2.0% (W/V)
Suspending agent 5~20%/(W/V)
Flocculating agent 1~2.5% (W/V)
Stabilizing agent 0.01~0.05% (W/V)
Antiseptic 0.01~0.6% (W/V)
Purified water 65~94% (V/V)
The percentage ratio of above composition is based on the cumulative volume meter of suspensoid.
Preparation method:
(1) earlier flocculating agent is dissolved in a small amount of purified water, adds principal agent deccox, wetting agent and surfactant again, stir mixture A;
(2) get an amount of purified water, add stabilizing agent and suspending agent, stir mixture B;
(3) mixture B is poured among the mixture A, and add antiseptic, fixed molten behind the stirring and evenly mixing to 100% (V/V) with purified water, obtain mixture C;
(4) mixture C is homogenized by high pressure homogenizer;
(5) by obtaining the deccox suspension behind twice circulation homogenizing.
According to the present invention, described principal agent deccox consumption is 3~30% (W/V).
Above-mentioned wetting agent includes but not limited to: a kind of or combination in ethanol, the propylene glycol.
Above-mentioned surfactant comprises but is not limited to: a kind of or combination of sodium laurylsulfate, dodecyl sodium sulfate, sorbitan mono-laurate, sorbitan monostearate, polyoxyethylene sorbitan monoleate, sorbitan monooleate, polyoxyethylene 20 sorbitan monooleate etc.
Above-mentioned flocculating agent includes but not limited to: citrate, citric acid hydrogen salt, tartrate, biatrate.
Above-mentioned stabilizing agent is a disodiumedetate.
Above-mentioned antiseptic includes but not limited to: a kind of or combination of phenol, methyl hydroxybenzoate, ethyl hydroxybenzoate, propylparaben etc.
Purpose of the present invention also provides the purposes of a kind of deccox suspension on the treatment chicken coccidiosis.
The specific embodiment
Embodiment 1:
Component:
Amounts of components
Deccox 6.0g
Ethanol 5.0ml
Sodium laurylsulfate 0.06g
Glycerol 10.0g
Sodium hydrogen citrate 2.0g
Disodiumedetate 0.05g
Phenol 0.50g
Methyl hydroxybenzoate 0.03g
Purified water 84ml
Collocation method:
(1) earlier sodium hydrogen citrate 2.0g is dissolved in the 10ml purified water, adds deccox 6.0g, ethanol 3.0ml and sodium laurylsulfate 0.06g again, stir mixture A.
(2) get the 60ml purified water again, add methyl hydroxybenzoate 0.03g, be placed to room temperature after the heating for dissolving, add disodiumedetate 0.05g and glycerol 8.0g again, stir mixture B.
(3) mixture B is poured among the mixture A, and add phenol 0.50g, behind the stirring and evenly mixing, add purified water 14ml again, obtain mixture C.
(4) mixture C is homogenized under the pressure of 100Mpa by high pressure homogenizer.
(5) by obtaining solid particle size deccox suspension between 5-500nm behind 2 circulation homogenizing.
Embodiment 2:
Component:
Amounts of components
Deccox 8.0g
Ethanol 5.0ml
Sodium laurylsulfate 0.1g
Glycerol 12.0g
Sodium hydrogen citrate 2.5g
Disodiumedetate 0.05g
Phenol 0.5g
Methyl hydroxybenzoate 0.03g
Purified water 82ml
Collocation method:
(1) earlier sodium hydrogen citrate 2.5g is dissolved in the 10ml purified water, adds deccox 8.0g, ethanol 5.0ml and sodium laurylsulfate 0.1g again, stir mixture A.
(2) get the 60ml purified water, add methyl hydroxybenzoate 0.03g, be placed to room temperature after the heating for dissolving, add disodiumedetate 0.05g and glycerol 8.0g again, stir mixture B.
(3) mixture B is poured among the mixture A, and add phenol 0.5g, behind the stirring and evenly mixing, add purified water 12ml again, obtain mixture C.
(4) mixture C is homogenized under the pressure of 60Mpa by high pressure homogenizer.
(5) by obtaining solid particle size deccox suspension between 100-1000nm behind 2 circulation homogenizing.
Embodiment 3:
Component:
Amounts of components
Deccox 30.0g
Ethanol 5.0ml
Dodecyl sodium sulfate 0.06g
Polyoxyethylene sorbitan monoleate 1.0g
Propylene glycol 5.0g
Glycerol 5.0g
Sodium carboxymethyl cellulose 2.0g
Sodium hydrogen citrate 2.0g
Disodiumedetate 0.05g
Phenol 0.5g
Methyl hydroxybenzoate 0.03g
Purified water 83ml
Collocation method:
(1) earlier sodium hydrogen citrate 2.0g is dissolved in the 10ml purified water, adds deccox 30g, ethanol 5.0ml and dodecyl sodium sulfate 0.06g again, stir mixture A.
(2) get the 60ml purified water, add methyl hydroxybenzoate 0.3g, be placed to room temperature after the heating for dissolving, add disodiumedetate 0.05g, sodium carboxymethyl cellulose 2.0g, glycerol 5.0g and propylene glycol 5.0g again, stir mixture B.
(3) mixture B is poured among the mixture A, and add phenol, behind the stirring and evenly mixing, add purified water 13ml again, obtain mixture C.
(4) mixture C is homogenized by high pressure homogenizer 60MPa.
(5) by obtaining solid particle size deccox suspension between 100-1000nm behind 2 circulation homogenizing.
Embodiment 4:
The stability test result:
1. accelerated tests is in order to measure this stability of formulation, requirement according to " veterinary drug stability test technical specification ", accelerated test under the ad hoc fixed following condition, product to embodiment 1, embodiment 2, embodiment 3 is tested, experimental condition: it is that 40 degrees centigrade, relative humidity are 75% calorstat that three kinds of deccox suspension samples of embodiment 1, embodiment 2, embodiment 3 are put into temperature, the sampling respectively in the 1st, 2,3,6 month, and observe character respectively and measure content, experimental result is as follows:
The deccox suspension of table 1 embodiment 1 quickens the stability test result
Inspection item |
Character |
The settling volume ratio |
Content |
Before the experiment |
Little yellow suspension |
0.96 |
6.10% |
One month |
Little yellow suspension |
0.95 |
6.08% |
Two months |
Little yellow suspension |
0.96 |
6.09% |
Three months |
Little yellow suspension |
0.96 |
6.07% |
Six months |
Little yellow suspension |
0.96 |
6.08% |
The deccox suspension of table 2 embodiment 2 quickens the stability test result
Inspection item |
Character |
The settling volume ratio |
Content |
Before the experiment |
Little yellow suspension |
0.98 |
8.21% |
One month |
Little yellow suspension |
0.98 |
8.18% |
Two months |
Little yellow suspension |
0.97 |
8.19% |
Three months |
Little yellow suspension |
0.98 |
8.17% |
Six months |
Little yellow suspension |
0.98 |
8.18% |
The deccox suspension of table 3 embodiment 3 quickens the stability test result
Inspection item |
Character |
The settling volume ratio |
Content |
Before the experiment |
Little yellow suspension |
0.94 |
30.5% |
One month |
Little yellow suspension |
0.94 |
30.2% |
Two months |
Little yellow suspension |
0.93 |
30.5% |
Three months |
Little yellow suspension |
0.93 |
30.1% |
Six months |
Little yellow suspension |
0.94 |
30.0% |
From assay, in the phase, character does not all change the deccox suspension of embodiment 1, embodiment 2, embodiment 3 at accelerated stability test, settling ratio illustrates that all greater than 0.9 dispersibility is fine, and content does not have significant change, every index all meets the requirements, and this product good stability is described.
Embodiment 5:
The clinical efficacy test
The clinical efficacy test
1 material
1.1 medicine embodiment 1 (deccox suspension); Deccox pre-mixing agent (content is 6%), producer provides by domestic production.
1.2 public young 200 of the brown egg of animal 1 age in days gold is raised in sterilization strictness on the net, free choice feeding and drinking-water, the conventional raising.Raising to 10 age in days is gathered stool examination, determines that non-ball worm infects.
1.3 a coccidiosis strain Eimeria tenella person of outstanding talent strain (E.tenellaHough ton st rain) is provided by Agricultural University Of Nanjing.
Coccidiosis is cultivated, the conventional reagent and the equipment of counting 1.4 reagent and equipment are used for.
2 methods
2.1 pilot study, determines that every chicken infects the coccidian oocyst number by trial test.Pre-test result is that every chicken infects 5 * 10
4Individual egg capsule.
2.2 180 of even, the healthy chickens of test grouping selective body weight average are divided into six groups of I, II, III, IV, V, VI, every group of 30 chickens are established 3 repetitions, and each repeats 10 chickens, all raises under the condition of non-ball worm.6 groups are respectively normal healthy controls group (not infecting not administration group), positive controls (infecting not administration group), deccox pre-mixing agent matched group (30mg/kg) and low dosage deccox suspension group (20mg/kg feedstuff), middle dosage deccox suspension group (30mg/kg feedstuff), high dose deccox suspension group (40mg/kg feedstuff).In entire test, the feedstuff of all groups and apparatus are all separately strict, in case the normal healthy controls group is subjected to coccidium infection.The administration of mixed feeding method is adopted in test.Feed to off-test continuously.Except that the normal healthy controls group, other are respectively organized chicken and drip clothes Eimeria tenella egg capsule (5 * 10 with the dropper per os when 14 ages in days
4Individual/only).
After 2.3 clinical observation is infected with record, observe for 3 times every day chicken health status, drink water and the situation of searching for food, morbidity and the death condition of chicken respectively organized in record, the promoting the circulation of blood of going forward side by side is just kept the score.In infecting the back the 7th day, all cut open test chicken extremely, observe and write down the caecum lesion of respectively organizing chicken, and calculate the pathological changes value.Calculate in the feces egg capsule output, calculate the cecal content egg sac number, and calculate the egg capsule value, calculate the anticoccidial index at last, come evaluate efficacy with this.
2.4 pharmacodynamics index
Average weight gain * 100% of group 2.4.1 the average weight gain of the relative weight gain rate the relative weight gain rate (%)=infection dispensing group/do not infect is not offerd medicine.
2.4.2 survival rate survival rate (%)=survival number of elements/chicken number * 100% of participating in the experiment
2.4.3 keeping the score, hemafecia is not with blood, the blood stool of+1 expression 25%, the blood stool of+2 expressions 50%, the blood stool of+3 expressions 75%, the blood stool of+4 expressions 100% by the feces of expression 100% in 0 fen.
2.4.4 the pathological changes value is kept the score to caecum lesion by the standard of keeping the score of Reid etc.Pathological changes value=every group of average lesion score * 10.Be 0~011 o'clock, the egg capsule value is 0; When the OPG value was 011~110, the egg capsule value was 1; When the OPG value was 210~510, the egg capsule value was 10; When the OPG value was 610~1010, the egg capsule value was 20; When OPG value 〉=1110, the egg capsule value is 40.
2.4.6 anticoccidial index (ACI) ACI=(rate of body weight gain+survival rate)-(pathological changes value+egg capsule value).When anti-ball index>180 are efficient; 160~180 is middle effect;<160 is poor efficiency;<120 is invalid.
3 results and analysis
3.1 anticoccidial effect is as shown in Table 1, the ACI value of middle dosage deccox suspension group (V group) is 199.6, than deccox pre-mixing agent group (III group) high 31.80%.The ACI value of high dose deccox suspension group (VI group) is 200.8, than deccox pre-mixing agent group (III group) high 33.00%.
3.2 gaining effect is as shown in Table 1, the relative weight gain rate of middle dosage deccox suspension group (V group) than deccox pre-mixing agent group (III group) high 9.3%; The relative weight gain rate of high dose deccox suspension group (VI group) than deccox pre-mixing agent group (III group) high 10.4%.
Compare with the deccox pre-mixing agent 3.3 analyze the deccox suspension of same dose, no matter the deccox suspension all has very big lifting than deccox pre-mixing agent effect on anticoccidial effect and gaining effect.
The test of pesticide effectiveness of table 1 pair chickling artificial challenge coccidian oocyst
Group |
Starting weight/g only
-1 |
End weight/g only
-1 |
Weightening finish/g only
-1 |
Rate of body weight gain/% |
Survival rate % |
The pathological changes value |
The egg capsule value |
Hemafecia is kept the score |
ACI |
I |
60.2±3.0 |
144.2±12.2 |
84.0±6.7 |
100.0 |
100.0 |
0 |
0 |
0 |
200.0 |
II |
59.7±2.4 |
116.7±14.9 |
57.0±7.6 |
67.9 |
63.3 |
34.2 |
42.0 |
3.3 |
55.0 |
III |
59.6±2.9 |
136.4±13.1 |
76.8±6.9 |
91.4 |
93.3 |
4.9 |
12.0 |
0.6 |
167.8 |
IV |
58.7±2.6 |
134.5±12.7 |
75.7±7.6 |
90.2 |
90.0 |
6.7 |
14.0 |
1.1 |
159.9 |
V |
59.2±2.5 |
143.8±14.6 |
84.6±8.3 |
100.7 |
100.0 |
1.0 |
0 |
0.1 |
199.6 |
VI |
58.9±2.9 |
144.4±15.7 |
85.5±7.2 |
101.8 |
100.0 |
1.0 |
0 |
0 |
200.8 |