CN104667280A - Immunomodulator for duck flavivirus immunotherapy as well as preparation method and application thereof - Google Patents
Immunomodulator for duck flavivirus immunotherapy as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to an immunomodulator for a duck flavivirus immunotherapy. The immunomodulator is a solution, and the solution comprises a solute and a solvent, wherein the solution is a sodium chloride aqueous solution with the mass percentage of 0.9%; and the solute is a mixture of a Toll-like receptor agonist, an NOD-like receptor agonist and a corrosion inhibitor. The invention also provides a preparation method and an application of the immunomodulator applied to the duck flavivirus immunotherapy. By adopting the immunomodulator for the duck flavivirus immunotherapy disclosed by the invention, the organism immunity of ducks can be quickly increased; and in addition, the immunomodulator is simple in preparation method and wide in application.
Description
Technical field
The present invention relates to a kind of immunomodulator and its preparation method and application, be specifically related to a kind of immunomodulator for duck flavivirus immunotherapy and its preparation method and application.
Background technology
Duck flavivirus (duck tembusu virus) be a kind of have a strong impact on laying ducks and plant duck to lay eggs the virus of function.Its clinical manifestation declines suddenly for function of laying eggs, and serious laying rate in 3-5 days reduces to zero by peak, feed intake also together with time decline, have the phenomena of mortality individually.Pathology analyses the serious change such as the hyperemia of visible ovary, swelling, necrosis and atrophy, and with phenomenons such as heating hyperpyrexias.For commonly encountered diseases viral disease, cellular immunization is the first line of defence of body, and after poisoning intrusion body, body can raise the secretion of some cytokine in cell aspect, thus causing the intracellular signaling between body immune system, activating immune system resists the intrusion of microorganism.Direct inhibitory action has copied to virus in some cytokine such as interleukin family and interferon family, so while pathogenic microorganism and body immune system carry out contacting, pathogenic microorganism can discharge or secrete the specific factor and suppress body produce the harmful cytokine of pathogenic microorganism thus cause affected animal hypoimmunity during disease.
After this disease was found first at home since 2010, cause Shandong, Hunan, province, Guangdong etc. 12 laying ducks drops in production over a large area, although Ge great research institution is all carrying out detailed basic research to this virus, some research institutions have have researched and developed the vaccine manufacturing experimently out this virus, as the application for a patent for invention file that publication number is CN102397539A, disclose a kind of Duckling flavivirus disease inactivated vaccine and preparation method thereof, this vaccine contains duck flavivirus strain through deactivation and adjuvant, its preparation method is as follows: the production of duck flavivirus strain kind of a poison is carried out large-scale culture, results virus liquid, by adding in adjuvant after the virus liquid deactivation of results, after emulsifying, namely obtain Duckling flavivirus disease inactivated vaccine, as the application for a patent for invention file that publication number is CN102757940A, disclose a kind of duck flavivirus and prepare the preparation method of inactivated oil-emulsion vaccine with this strain, described duck flavivirus is duck flavivirus AH-F10 strain, deposit number is: CCTCC V201213, by clinical separation and purification virus, by the virus liquid after purification through formalin-inactivated, make duck flavivirus oil-emulsion bacterin with after oil emulsion adjuvant emulsifying.
Above-mentioned vaccine can make laying ducks and plant duck plays good prevention infection effect to tooth banzi virus, but traditional vaccine potency is strong not, make the cycle of duck raising immunity of organisms longer, and when disease produces, vaccinate can not play a role, the burden of duck body can be increased the weight of on the contrary, and in advance vaccinate, work continue numerous and diverse, take time and effort, the cost of duck cultivation can be increased.
Summary of the invention
The object of the present invention is to provide a kind of immunomodulator for duck flavivirus immunotherapy that can suppress the copying of duck flavivirus, fast lifting duck body immunity, the present invention simultaneously also provides the preparation method and application of this immunomodulator.
For solving the problem, the technical solution adopted in the present invention is as follows:
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described solution comprises solute and solvent, wherein, and the sodium-chloride water solution of described solution to be mass percent be 0.7-1%; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter;
Described Toll-like receptor agonist is the one or more kinds of combinations in TLR1 agonist, TLR2 agonist, TLR3 agonist, TLR4 agonist, TLR5 agonist, TLR6 agonist, TLR7 agonist, TLR8 agonist, TLR9 agonist, TLR10 agonist, TLR11 agonist, TLR12 agonist and TLR13 agonist, and the concentration of described Toll-like receptor agonist is 0.001-20 μ g/ml;
Described NOD sample receptor stimulating agent is the one or more kinds of combinations in NOD1 receptor stimulating agent, NOD2 receptor stimulating agent, NLRP3 receptor stimulating agent, NLRC4 receptor stimulating agent, NALP1 receptor stimulating agent and NLRP6 receptor stimulating agent, and the concentration of described NOD sample receptor stimulating agent is 0.001-15 μ g/ml;
Described slow releasing agent is the one or more kinds of combinations in carbomer, PVPK, cyclodextrin and sodium alginate, and the percentage by volume of described slow releasing agent is 0.01-1.5%.
In the present invention, preferred scheme is the concentration of described Toll-like receptor agonist is 0.05-0.5 μ g/ml, the concentration of described NOD sample receptor stimulating agent is 0.01-0.1 μ g/ml, described slow releasing agent is the combination of PVPK and sodium alginate, the percentage by volume of described PVPK is 0.05-0.5%, and the percentage by volume of described sodium alginate is 0.01-1%.
In the present invention, the combination of preferred scheme to be described NOD sample receptor stimulating agent be NOD1 receptor stimulating agent and NOD2 receptor stimulating agent; Described Toll-like receptor agonist is:
The combination of TLR2 agonist, TLR4 agonist, TLR3 agonist and TLR7 agonist;
Or, the combination of TLR2 agonist, TLR4 agonist, TLR8 agonist and TLR9 agonist;
Or, the combination of TLR2 agonist, TLR4 agonist, TLR-3 agonist and TLR9 agonist.
In the present invention, preferred scheme is the concentration of described Toll-like receptor agonist is 0.001-0.01 μ g/ml, the concentration of described NOD sample receptor stimulating agent is 0.001-0.01 μ g/ml, described slow releasing agent is the combination of PVPK and carbomer, the percentage by volume of described PVPK is 0.05-0.5%, and the percentage by volume of described sodium alginate is 0.01-0.1%.
In the present invention, the combination of preferred scheme to be described NOD sample receptor stimulating agent be NOD1 receptor stimulating agent and NOD2 receptor stimulating agent; Described Toll-like receptor agonist is:
The combination of TLR2 agonist, TLR4 agonist, TLR3 agonist and TLR7 agonist;
Or, the combination of TLR2 agonist, TLR4 agonist, TLR8 agonist and TLR9 agonist;
Or, the combination of TLR2 agonist, TLR4 agonist, TLR-3 agonist and TLR9 agonist.
In the present invention, preferred scheme to be described mass percent be 0.9% sodium-chloride water solution be sterile NaCl aqueous solution.
Also provide this for the preparation method of the immunomodulator of duck flavivirus immunotherapy in the present invention: to weigh Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter according to formula ratio, then mass percent Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter being added formula ratio is in the sodium-chloride water solution of 0.9%, be stirred to dissolving, aseptic filtration, collect filtrate, obtain product.
In the present invention, preferred scheme is the rotating speed of described stirring is 100-500r/min, and the filter membrane that described aseptic filtration adopts is 0.1-0.45 μm.
Immunomodulator for duck flavivirus immunotherapy preparation of the present invention is as the application of nasal solutions.
In the present invention, also provide a kind of application of immunomodulator as injection utilizing duck flavivirus immunotherapy of the present invention.
Compared with prior art, the invention has the advantages that: adopt the immunomodulator for duck flavivirus immunotherapy of the present invention, can use between period of disease and can improve immunity of organisms and some specific cytokine fast, suppress copying of virus, thus improve laying ducks autoimmunity, to antiviral infringement; In addition, the preparation method of this immunomodulator is simple, easy to operate; This immunomodulator as nasal drop and injection etc., can be widely used.
Detailed description of the invention
In all embodiments of the present invention, this immunomodulator being used for duck flavivirus immunotherapy is prepared according to following method: weighed Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter according to formula ratio, then mass percent Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter being added formula ratio is in the sodium-chloride water solution of 0.9%, be stirred to dissolving, aseptic filtration, the rotating speed of described stirring is 200r/min, the filter membrane that described aseptic filtration adopts is 0.25 μm, collect filtrate, obtain product.
Embodiment 1
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sterile NaCl aqueous solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is: the combination of TLR2 agonist, TLR4 agonist, TLR3 agonist and TLR7 agonist, the concentration of described Toll-like receptor agonist is 0.05 μ g/ml (wherein, the concentration of the concentration of TLR2 agonist to be the concentration of 0.01 μ g/ml, TLR4 agonist be 0.01 μ g/ml, TLR3 agonist is the concentration of 0.02 μ g/ml, TLR7 agonist is 0.01 μ g/ml); Described NOD sample receptor stimulating agent is the combination for NOD1 receptor stimulating agent and NOD2 receptor stimulating agent, the concentration of described NOD sample receptor stimulating agent is 0.01 μ g/ml (wherein, the concentration of NOD1 receptor stimulating agent is the concentration of 0.006 μ g/ml, NOD2 receptor stimulating agent is 0.004 μ g/ml); Described slow releasing agent is the combination of PVPK and carbomer, and the percentage by volume of described PVPK is 0.05%, and the percentage by volume of described sodium alginate is 0.01%.
Embodiment 2
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sterile NaCl aqueous solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is the combination of TLR2 agonist, TLR4 agonist, TLR8 agonist and TLR9 agonist, the concentration of described Toll-like receptor agonist is 0.5 μ g/ml (wherein, the concentration of the concentration of TLR2 agonist to be the concentration of 0.2 μ g/ml, TLR4 agonist be 0.1 μ g/ml, TLR8 agonist is the concentration of 0.1 μ g/ml, TLR9 agonist is 0.1 μ g/ml); Described NOD sample receptor stimulating agent is the combination of NOD1 receptor stimulating agent and NOD2 receptor stimulating agent, the concentration of described NOD sample receptor stimulating agent is 0.1 μ g/ml (wherein, the concentration of NOD1 receptor stimulating agent is the concentration of 0.05 μ g/ml, NOD2 receptor stimulating agent is 0.05 μ g/ml); Described slow releasing agent is the combination of PVPK and carbomer, and the percentage by volume of described PVPK is 0.5%, and the percentage by volume of described sodium alginate is 0.1%.
Embodiment 3
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sodium-chloride water solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is the combination of TLR2 agonist, TLR4 agonist, TLR3 agonist and TLR9 agonist, the concentration of described Toll-like receptor agonist is 0.2 μ g/ml (wherein, the concentration of the concentration of TLR2 agonist to be the concentration of 0.01 μ g/ml, TLR4 agonist be 0.07 μ g/ml, TLR3 agonist is the concentration of 0.06 μ g/ml, TLR9 agonist is 0.06 μ g/ml); Described NOD sample receptor stimulating agent is the combination of NOD1 receptor stimulating agent and NOD2 receptor stimulating agent, the concentration of described NOD sample receptor stimulating agent is 0.05 μ g/ml (wherein, the concentration of NOD1 receptor stimulating agent is the concentration of 0.04 μ g/ml, NOD2 receptor stimulating agent is 0.01 μ g/ml); Described slow releasing agent is the combination of PVPK and carbomer, and the percentage by volume of described PVPK is 0.3%, and the percentage by volume of described sodium alginate is 0.06%.
Embodiment 4
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sodium-chloride water solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is the combination of TLR2 agonist, TLR4 agonist, TLR3 agonist and TLR7 agonist, the concentration of Toll-like receptor agonist is 0.001 μ g/ml (wherein, the concentration of the concentration of TLR2 agonist to be the concentration of 0.0002 μ g/ml, TLR4 agonist be 0.0003 μ g/ml, TLR3 agonist is the concentration of 0.0004 μ g/ml, TLR7 agonist is 0.0001 μ g/ml); Described NOD sample receptor stimulating agent is the combination of NOD1 receptor stimulating agent (0.008 μ g/ml) and NOD2 receptor stimulating agent (0.002 μ g/ml), the concentration of described NOD sample receptor stimulating agent is 0.01 μ g/ml (wherein, the concentration of NOD1 receptor stimulating agent is the concentration of 0.008 μ g/ml, NOD2 receptor stimulating agent is 0.002 μ g/ml); Described slow releasing agent is the combination of PVPK and carbomer, and the percentage by volume of described PVPK is 0.01%, and the percentage by volume of described sodium alginate is 0.1%.
Embodiment 5
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sodium-chloride water solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is the combination of TLR2 agonist, TLR4 agonist, TLR8 agonist and TLR9 agonist, the concentration of Toll-like receptor agonist is 0.01 μ g/ml (wherein, the concentration of the concentration of TLR2 agonist to be the concentration of 0.003 μ g/ml, TLR4 agonist be 0.004 μ g/ml, TLR8 agonist is the concentration of 0.001 μ g/ml, TLR9 agonist is 0.002 μ g/ml); Described NOD sample receptor stimulating agent is the combination of NOD1 receptor stimulating agent and NOD2 receptor stimulating agent, the concentration of described NOD sample receptor stimulating agent is 0.001 μ g/ml (wherein, the concentration of NOD1 receptor stimulating agent is the concentration of 0.0002 μ g/ml, NOD2 receptor stimulating agent is 0.008 μ g/ml); Described slow releasing agent is the combination of PVPK and carbomer, and the percentage by volume of described PVPK is 1%, and the percentage by volume of described sodium alginate is 0.01%.
Embodiment 6
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sodium-chloride water solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is the combination of TLR2 agonist, TLR4 agonist, TLR3 agonist and TLR9 agonist, the concentration of Toll-like receptor agonist is 0.008 μ g/ml (wherein, the concentration of the concentration of TLR2 agonist to be the concentration of 0.002 μ g/ml, TLR4 agonist be 0.0003 μ g/ml, TLR3 agonist is the concentration of 0.002 μ g/ml, TLR9 agonist is 0.001 μ g/ml); Described NOD sample receptor stimulating agent is the combination of NOD1 receptor stimulating agent and NOD2 receptor stimulating agent, the concentration of described NOD sample receptor stimulating agent is 0.006 μ g/ml (wherein, the concentration of NOD1 receptor stimulating agent is the concentration of 0.002 μ g/ml, NOD1 receptor stimulating agent is 0.004 μ g/ml); Described slow releasing agent is the combination of PVPK and carbomer, and the percentage by volume of described PVPK is 0.5%, and the percentage by volume of described sodium alginate is 0.06%.
Experimental example 1
Laying ducks field, Guangdong cultivation laying ducks 1500, egg-laying peak can reach 9.5 one-tenth.To enter behind peak period 2 days, this batch of laying ducks have 5-8 only to occur every day the mental status is depressed, inappetence, egg drop reduction.This situation of self-discovery is after 5 days, and full group's laying ducks egg production obviously declines, and falls to 4 one-tenth eggs by original 9.5 one-tenth eggs, and feed intake sharply declines 30% of the feed intake reducing to original.Pathology analyses visible ovary hyperemia in various degree, and part laying ducks ovary blackening is downright bad; Adopt ovary tissue and carry out laboratory polymerase chain formula reaction diagnosis, detect and be diagnosed as duck flavivirus.
The immunomodulator being used for duck flavivirus immunotherapy of embodiment 1-3 is applied as nasal drop, the immunomodulator being used for duck flavivirus immunotherapy of embodiment 4-6 is applied as injection; This group of laying ducks are divided into first group and second group, every group each 750, are then divided into three groups by every group, often organize each 250 (being respectively first group to the 6th group, corresponding with embodiment 1-6 respectively); Wherein first group is carried out collunarium (corresponding 3rd group of corresponding first group of embodiment 1, corresponding second group of embodiment 2, embodiment 3) with above-mentioned immunomodulator, and every day 1 time, every 1-2 drips at every turn; Second group of above-mentioned immunomodulator of group carries out injecting (corresponding 4th group of embodiment 4, corresponding 5th group of embodiment 5, corresponding 6th group of embodiment 6), every day 1 time, each every only 0.5 milliliter.Result of use is observed every 4 hours.
Result: injection group (namely second group, corresponding embodiment 4-6) use after second day feed intake start to rise, the rising of collunarium group (namely first group, corresponding embodiment 1-3) feed intake, but time on slightly sky similar to injection group.Continuous use is after three days, and feed intake returns to original 70%, and egg production maintains about 4 one-tenth and do not change.Continuous Tracking is after one week, and feed intake returns to normally, and egg production returns to 8 one-tenth.
Embodiment 7-13
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described immunomodulator is that solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sterile NaCl aqueous solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is the one or more kinds of combinations in TLR1 agonist, TLR2 agonist, TLR3 agonist, TLR4 agonist, TLR5 agonist, TLR6 agonist, TLR7 agonist, TLR8 agonist, TLR9 agonist, TLR10 agonist, TLR11 agonist, TLR12 agonist and TLR13 agonist; Described NOD sample receptor stimulating agent is the one or more kinds of combinations in NOD1 receptor stimulating agent, NOD2 receptor stimulating agent, NLRP3 receptor stimulating agent, NLRC4 receptor stimulating agent, NALP1 receptor stimulating agent and NLRP6 receptor stimulating agent; Described slow releasing agent is the one or more kinds of combinations in carbomer, PVPK, cyclodextrin and sodium alginate.
The concrete component of the Toll-like receptor agonist in described solute, NOD sample receptor stimulating agent and corrosion inhibiter and consumption see the following form 1, and (wherein the unit of Toll-like receptor agonist, NOD sample receptor stimulating agent is μ g/ml, the unit of slow releasing agent is percentage by volume, and "--" expression is not added):
Table 1: the solute formula table implementing the immunomodulator of 7-13
Experimental example 2
Laying ducks field, Guangzhou cultivation laying ducks 1400, egg-laying peak can reach 9.5 one-tenth.To enter behind peak period 2 days, this batch of laying ducks have 5-8 only to occur every day the mental status is depressed, inappetence, egg drop reduction.This situation of self-discovery is after 5 days, and full group's laying ducks egg production obviously declines, and falls to 4 one-tenth eggs by original 9.5 one-tenth eggs, and feed intake sharply declines 30% of the feed intake reducing to original.Pathology analyses visible ovary hyperemia in various degree, and part laying ducks ovary blackening is downright bad; Adopt ovary tissue and carry out laboratory polymerase chain formula reaction diagnosis, detect and be diagnosed as duck flavivirus.
The immunomodulator being used for duck flavivirus immunotherapy of embodiment 7-10 is applied as nasal drop, the immunomodulator being used for duck flavivirus immunotherapy of embodiment 11-13 is applied as injection; This group of laying ducks are divided into seven groups, are respectively the 7th group to the 13 group, often organize each 200 (corresponding with embodiment 7-13 respectively); Wherein the 7th group to the tenth group is carried out collunarium (corresponding 7th group of embodiment 7, corresponding 8th group of embodiment 8, corresponding 9th group of embodiment 9, corresponding tenth group of embodiment 10) with above-mentioned immunomodulator, and every day 1 time, every 1-2 drips at every turn; 11 group to the 13 group is carried out injecting (corresponding 11 group of embodiment 11, corresponding 12 group of embodiment 12, corresponding 13 group of embodiment 13) with above-mentioned immunomodulator, every day 1 time, each every only 0.5 milliliter.Result of use is observed every 4 hours.
Result: injection group (corresponding embodiment 11-13) use after the 3rd day feed intake start to rise, collunarium group (corresponding embodiment 7-10) feed intake rises similar to injection group but on the time two days a little later.Continuous use is after five days, and feed intake returns to original 70%, and egg production maintains about 4 one-tenth and do not change.Continuous Tracking is after 10 days, and feed intake returns to normally, and egg production returns to 8 one-tenth.
Embodiment 14-20
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described immunomodulator is that solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sterile NaCl aqueous solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is the one or more kinds of combinations in TLR1 agonist, TLR2 agonist, TLR3 agonist, TLR4 agonist, TLR5 agonist, TLR6 agonist, TLR7 agonist, TLR8 agonist, TLR9 agonist, TLR10 agonist, TLR11 agonist, TLR12 agonist and TLR13 agonist; Described NOD sample receptor stimulating agent is the one or more kinds of combinations in NOD1 receptor stimulating agent, NOD2 receptor stimulating agent, NLRP3 receptor stimulating agent, NLRC4 receptor stimulating agent, NALP1 receptor stimulating agent and NLRP6 receptor stimulating agent; Described slow releasing agent is the one or more kinds of combinations in carbomer, PVPK, cyclodextrin and sodium alginate.
The concrete component of the Toll-like receptor agonist in described solute, NOD sample receptor stimulating agent and corrosion inhibiter and consumption see the following form 2, and (wherein the unit of Toll-like receptor agonist, NOD sample receptor stimulating agent is μ g/ml, the unit of slow releasing agent is percentage by volume, and "--" expression is not added):
Table 2: the solute formula table implementing the immunomodulator of 14-20
Experimental example 3
Laying ducks field, Guangzhou cultivation laying ducks 1400, egg-laying peak can reach 9.5 one-tenth.To enter behind peak period 2 days, this batch of laying ducks have 5-8 only to occur every day the mental status is depressed, inappetence, egg drop reduction.This situation of self-discovery is after 5 days, and full group's laying ducks egg production obviously declines, and falls to 4 one-tenth eggs by original 9.5 one-tenth eggs, and feed intake sharply declines 30% of the feed intake reducing to original.Pathology analyses visible ovary hyperemia in various degree, and part laying ducks ovary blackening is downright bad; Adopt ovary tissue and carry out laboratory polymerase chain formula reaction diagnosis, detect and be diagnosed as duck flavivirus.
The immunomodulator being used for duck flavivirus immunotherapy of embodiment 14-16 is applied as nasal drop, the immunomodulator being used for duck flavivirus immunotherapy of embodiment 17-20 is applied as injection; This group of laying ducks are divided into seven groups, are respectively the 14 group to the 20 group, often organize each 200 (corresponding with embodiment 14-20 respectively); Wherein the 14 group to the 16 group is carried out collunarium (corresponding 14 group of embodiment 14, corresponding 15 group of embodiment 15, corresponding 16 group of embodiment 16) with above-mentioned immunomodulator, and every day 1 time, every 1-2 drips at every turn; 17 group to the 20 group is carried out injecting (corresponding 17 group of embodiment 17, corresponding 18 group of embodiment 18, corresponding 19 group of embodiment 19, corresponding 20 group of embodiment 19) with above-mentioned immunomodulator, every day 1 time, each every only 0.5 milliliter.Result of use is observed every 4 hours.
Result: injection group (corresponding embodiment 17-20) use after the 4th day feed intake start to rise, collunarium group (corresponding embodiment 14-16) feed intake rises similar to injection group but on the time 2 days a little later.Continuous use is after 4 days, and feed intake returns to original 70%, and egg production maintains about 4 one-tenth and do not change.Continuous Tracking is after 9 weeks, and feed intake returns to normally, and egg production returns to 8 one-tenth.
Embodiment 21-27
For an immunomodulator for duck flavivirus immunotherapy, described immunomodulator is solution, and described immunomodulator is that solution comprises solute and solvent, wherein, described solution to be mass percent be 0.9% sterile NaCl aqueous solution; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter; Described Toll-like receptor agonist is the one or more kinds of combinations in TLR1 agonist, TLR2 agonist, TLR3 agonist, TLR4 agonist, TLR5 agonist, TLR6 agonist, TLR7 agonist, TLR8 agonist, TLR9 agonist, TLR10 agonist, TLR11 agonist, TLR12 agonist and TLR13 agonist; Described NOD sample receptor stimulating agent is the one or more kinds of combinations in NOD1 receptor stimulating agent, NOD2 receptor stimulating agent, NLRP3 receptor stimulating agent, NLRC4 receptor stimulating agent, NALP1 receptor stimulating agent and NLRP6 receptor stimulating agent; Described slow releasing agent is the one or more kinds of combinations in carbomer, PVPK, cyclodextrin and sodium alginate.
The concrete component of the Toll-like receptor agonist in described solute, NOD sample receptor stimulating agent and corrosion inhibiter and consumption see the following form 3, and (wherein the unit of Toll-like receptor agonist, NOD sample receptor stimulating agent is μ g/ml, the unit of slow releasing agent is percentage by volume, and "--" expression is not added):
Table 3: the solute formula table implementing the immunomodulator of 21-27
Experimental example 4
Laying ducks field, Guangzhou cultivation laying ducks 1400, egg-laying peak can reach 9.5 one-tenth.To enter behind peak period 2 days, this batch of laying ducks have 5-8 only to occur every day the mental status is depressed, inappetence, egg drop reduction.This situation of self-discovery is after 5 days, and full group's laying ducks egg production obviously declines, and falls to 4 one-tenth eggs by original 9.5 one-tenth eggs, and feed intake sharply declines 30% of the feed intake reducing to original.Pathology analyses visible ovary hyperemia in various degree, and part laying ducks ovary blackening is downright bad; Adopt ovary tissue and carry out laboratory polymerase chain formula reaction diagnosis, detect and be diagnosed as duck flavivirus.
The immunomodulator being used for duck flavivirus immunotherapy of embodiment 21-23 is applied as nasal drop, the immunomodulator being used for duck flavivirus immunotherapy of embodiment 24-27 is applied as injection; This group of laying ducks are divided into seven groups, are respectively the 21 group to the 27 group, often organize each 200 (corresponding with embodiment 21-27 respectively); Wherein the 21 group to the 23 group is carried out collunarium (corresponding 21 group of embodiment 21, corresponding 22 group of embodiment 22, corresponding 23 group of embodiment 23) with above-mentioned immunomodulator, and every day 1 time, every 1-2 drips at every turn; 24 group to the 27 group is carried out injecting (corresponding 24 group of embodiment 24, corresponding 25 group of embodiment 25, corresponding 26 group of embodiment 26, corresponding 27 group of embodiment 27) with above-mentioned immunomodulator, every day 1 time, each every only 0.5 milliliter.Result of use is observed every 4 hours.
Result: injection group (corresponding embodiment 24-27) use after the 4th day feed intake start to rise, collunarium group (corresponding embodiment 21-23) feed intake rises similar to injection group but on the time 3 days a little later.Continuous use is after 6 days, and feed intake returns to original 70%, and egg production maintains about 4 one-tenth and do not change.Continuous Tracking is after 10 days, and feed intake returns to normally, and egg production returns to 8 one-tenth.
Above-mentioned embodiment is only the preferred embodiment of the present invention; can not limit the scope of protection of the invention with this, change and the replacement of any unsubstantiality that those skilled in the art does on basis of the present invention all belong to the present invention's scope required for protection.
Claims (10)
1. for an immunomodulator for duck flavivirus immunotherapy, it is characterized in that: described immunomodulator is solution, and described solution comprises solute and solvent, wherein, the sodium-chloride water solution of described solution to be mass percent be 0.7-1%; Described solute is the mixture of Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter;
Described Toll-like receptor agonist is the one or more kinds of combinations in TLR1 agonist, TLR2 agonist, TLR3 agonist, TLR4 agonist, TLR5 agonist, TLR6 agonist, TLR7 agonist, TLR8 agonist, TLR9 agonist, TLR10 agonist, TLR11 agonist, TLR12 agonist and TLR13 agonist, and the concentration of described Toll-like receptor agonist is 0.001-20 μ g/ml;
Described NOD sample receptor stimulating agent is the one or more kinds of combinations in NOD1 receptor stimulating agent, NOD2 receptor stimulating agent, NLRP3 receptor stimulating agent, NLRC4 receptor stimulating agent, NALP1 receptor stimulating agent and NLRP6 receptor stimulating agent, and the concentration of described NOD sample receptor stimulating agent is 0.001-15 μ g/ml;
Described slow releasing agent is the one or more kinds of combinations in carbomer, PVPK, cyclodextrin and sodium alginate, and the percentage by volume of described slow releasing agent is 0.01-1.5%.
2. the immunomodulator for duck flavivirus immunotherapy according to claim 1, it is characterized in that: the concentration of described Toll-like receptor agonist is 0.05-0.5 μ g/ml, the concentration of described NOD sample receptor stimulating agent is 0.01-0.1 μ g/ml, described slow releasing agent is the combination of PVPK and sodium alginate, the percentage by volume of described PVPK is 0.05-0.5%, and the percentage by volume of described sodium alginate is 0.01-1%.
3. the immunomodulator for duck flavivirus immunotherapy according to claim 2, is characterized in that: described NOD sample receptor stimulating agent is the combination of NOD1 receptor stimulating agent and NOD2 receptor stimulating agent; Described Toll-like receptor agonist is:
The combination of TLR2 agonist, TLR4 agonist, TLR3 agonist and TLR7 agonist;
Or, the combination of TLR2 agonist, TLR4 agonist, TLR8 agonist and TLR9 agonist;
Or, the combination of TLR2 agonist, TLR4 agonist, TLR-3 agonist and TLR9 agonist.
4. the immunomodulator for duck flavivirus immunotherapy according to claim 1, it is characterized in that: the concentration of described Toll-like receptor agonist is 0.001-0.01 μ g/ml, the concentration of described NOD sample receptor stimulating agent is 0.001-0.01 μ g/ml, described slow releasing agent is the combination of PVPK and carbomer, the percentage by volume of described PVPK is 0.01-1%, and the percentage by volume of described sodium alginate is 0.01-0.1%.
5. the immunomodulator for duck flavivirus immunotherapy according to claim 4, is characterized in that: described NOD sample receptor stimulating agent is the combination of NOD1 receptor stimulating agent and NOD2 receptor stimulating agent; Described Toll-like receptor agonist is:
The combination of TLR2 agonist, TLR4 agonist, TLR3 agonist and TLR7 agonist;
Or, the combination of TLR2 agonist, TLR4 agonist, TLR8 agonist and TLR9 agonist;
Or, the combination of TLR2 agonist, TLR4 agonist, TLR-3 agonist and TLR9 agonist.
6. the immunomodulator for duck flavivirus immunotherapy according to any one of claim 1-5: it is characterized in that: described mass percent be 0.9% sodium-chloride water solution be sterile NaCl aqueous solution.
7. the preparation method of the immunomodulator for duck flavivirus immunotherapy according to claim 1, it is characterized in that: weighed Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter according to formula ratio, then mass percent Toll-like receptor agonist, NOD sample receptor stimulating agent and corrosion inhibiter being added formula ratio is in the sodium-chloride water solution of 0.9%, be stirred to dissolving, aseptic filtration, collect filtrate, obtain product.
8. the preparation method of the immunomodulator for duck flavivirus immunotherapy according to claim 7, is characterized in that: the rotating speed of described stirring is 100-500r/min, and the filter membrane that described aseptic filtration adopts is 0.1-0.45 μm.
9. according to the immunomodulator for duck flavivirus immunotherapy of any one of claim 1-5 as the application of nasal solutions.
10. according to the immunomodulator for duck flavivirus immunotherapy of any one of claim 1-5 as the application of injection.
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| CN103608036A (en) * | 2011-06-19 | 2014-02-26 | 瓦克西尼私人有限公司 | Vaccine adjuvant composition comprising inulin particles |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103608036A (en) * | 2011-06-19 | 2014-02-26 | 瓦克西尼私人有限公司 | Vaccine adjuvant composition comprising inulin particles |
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