Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
  • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
  • C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/12—Antidiarrhoeals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
  • C07K2317/11—Immunoglobulins specific features characterized by their source of isolation or production isolated from eggs
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
  • C07K2317/23—Immunoglobulins specific features characterized by taxonomic origin from birds

Definitions

• the present invention relates to a method for administering an egg produced by a bird immunized with a parvovirus or an antigen derived from a parvovirus, an egg-treated product thereof, an anti-parvovirus infection composition and a pet food containing the same, and an anti-parvovirus antibody.
• the present invention relates to a method for treating or preventing Parpovirus infection.
• Diarrhea is one of the most common diseases in humans, livestock and pets, but can cause dehydration and other complications in young individuals, especially in the suckling period, and can be severe.
• the causes of diarrhea are divided into those that are transmitted by viruses and bacteria and those that are not transmitted by the environment, stress, and diet.Infectious diarrhea is transmitted to other individuals and spreads throughout the population. Often causes enormous damage.
• the major pathogens of infectious diarrhea include viruses and bacteria. These pathogens develop when they enter the intestinal tract orally with food or beverages and colonize and grow on the intestinal mucosa. It is not uncommon for two or more pathogens to become infected together and make the condition more serious. Parvovirus infection is one of the problems that has become a problem with such viral infections.
• Parvovirus infections can cause severe symptoms such as a rapid rise in body temperature, diarrhea, vomiting, and loss of appetite.
• the white blood cell count (WBC) also decreases sharply.
• the canine parpovirus is a very resistant virus that survives for several months to several years in a normal environment, and there are many cases of spontaneous onset. Especially when bred as a group, they can be violent, and death from this infectious disease at breeder pet shops has become a major problem.
• Vaccines against parvovirus infection vaccines have had problems with their protective effects for some time. Intestinal sensation It is important to produce IgA antibodies to protect against infectious diseases.Inactivated vaccines currently used produce IgG antibodies when injected intramuscularly, but produce IgA antibodies in the intestinal tract. Since the amount is small, it is difficult to obtain the desired effect. On the other hand, when a live vaccine is administered orally, a large amount of IgA antibody is produced and a protective effect is observed, so the live vaccine is mainly used for viral diarrhea, but its effect is not sufficient. I can't say. In addition, vaccines cannot prevent the onset of infection due to childhood until vaccine administration can be started, and thus are not useful in countermeasures against the onset of childhood.
• chicken egg antibodies are known as antibodies suitable for oral administration.
• Pharmaceutical compositions using hen egg antibodies include the 987P, K88 and K99 antigens of enterotoxigenic Escherichia coli causing porcine colibacillosis, and the K99 antigens of enterotoxigenic Escherichia coli causing cholesterol colibacillosis.
• a polyclonal antibody specific to the antigen, obtained by immunizing a chicken with any one or more antigens and recovering the antibody from at least the portion of the egg produced by the immunized chicken that contains the yolk Oral prophylactic and therapeutic agents for colibacillosis are known (see Patent No. 2034005).
• compositions for swine epidemic diarrhea virus infection milk or milk components thereof collected from cattle immunized with swine epidemic diarrhea virus, and egg yolk or egg collected from chickens immunized with the virus
• a pharmaceutical composition containing an antibody is known (see JP-A-10-265393).
• an anti-parvovirus infection composition containing an antibody obtained from egg yolk of a chicken immunized with canine parvovirus is known (Japanese Patent Application Laid-Open No. -259462).
• An object of the present invention is to effectively prevent the onset of parvovirus infection in pets, And to provide an anti-parpovirus infection composition and pet food that is effective for the prevention and treatment of parvovirus infection, effectively improving symptoms such as diarrhea, vomiting and weight loss specific to parvovirus infection. is there.
• the present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, an egg obtained by immunizing birds with an antigen derived from parvovirus or parvovirus or a processed product thereof, which has a high neutralizing antibody titer
• the present inventors have found that the above-mentioned problems can be solved by the present invention, and have completed the present invention.
• the anti-parpovirus antibody contained in eggs obtained by immunizing birds using the parvovirus as an antigen or in a processed product thereof eliminates the pathogenicity by attaching to the pathogen and neutralizing the pathogen. Alternatively, it may abolish the pathogenicity by attaching to the cell adhesion factor of the pathogen and preventing the pathogen from attaching to intestinal cells.
• the present invention includes the following inventions.
• An anti-Parpovirus composition comprising the egg or the processed egg product according to any one of (1) to (3).
• a pet food comprising the egg or the processed egg product according to any one of (1) to (3).
• a method for treating or preventing parvovirus infection by administering to a non-human animal an anti-parvovirus antibody obtained from eggs produced by birds immunized with parvovirus or an antigen derived from parvovirus.
• immunization with an antigen derived from parvovirus or parvovirus is performed.
• birds include, but are not limited to, chickens and quail. From the viewpoint of mass production of antibodies, it is preferable to use chickens, particularly egg-laying species.
• examples of parvovirus strains that can be used for immunization of birds include CPK-2, Cp83016, Kawazoe, and the like. In particular, it is preferable to use CPK-2.
• CPK-2 Cp83016
• These parvovirus strains can be obtained from the Ministry of Agriculture, Forestry and Fisheries Animal Health Laboratory. By immunizing birds with the parvovirus strain as described above, eggs and processed egg products having a high neutralizing antibody titer against parvovirus can be obtained.
• an antigen derived from parvovirus means a peptide composed of a part of a component of parvovirus, and includes, for example, a parvovirus capsid, a nucleocapsid, and a capsomere. Also included are those consisting of parts. In the present invention, it is preferable to use such an antigen derived from parvovirus.
• Parvovirus and antigens derived therefrom are contained in virus lysates and the like, and both purified and unpurified ones can be used, but it is preferable to use purified ones.
• ordinary methods such as ammonium sulfate precipitation, column chromatography, and electrophoresis can be used alone or in combination of two or more.
• the column chromatography may use one kind of column, or may combine two or more kinds of columns.
• ion exchange chromatography, gel filtration chromatography, reverse phase chromatography, affinity column chromatography, etc. are used.
• adjuvants such as Freund's complete adjuvant (FCA) and Freund's incomplete adjuvant (FIA) can be used as necessary.
• FCA Freund's complete adjuvant
• FIA Freund's incomplete adjuvant
• Immunization is mainly performed by intravenous, subcutaneous, intramuscular, or intraperitoneal injection, but it can also be performed by oral administration, nasal administration, eye drops, or the like.
• the immunization interval is not particularly limited, and immunization is performed 1 to 10 times at intervals of several days to several weeks. Usually, within a few weeks of the first immunization, antibodies specifically reacting with the administered antigen are obtained in eggs, particularly yolk.
• the amount of antigen to be inoculated is the amount of protein, and 0.01 to:!
• the antibody titer in the egg can be measured by enzyme-linked immunosorbent assay (ELISA), radioimmunoassay, agglutination
• ELISA enzyme-linked immunosorbent assay
• the antibody titer can be measured using a body method or a neutralization reaction or the like, and the transition of the antibody titer can be tracked by measuring the antibody titer at intervals of about two weeks after immunization. Usually, a high antibody titer can be obtained for about 4 months. If the antibody titer decreases after immunization, the antibody titer can be kept high by performing additional immunization at appropriate intervals.
• the processed egg product is not particularly limited as long as it contains an antibody against Parpovirus used for immunization of birds as an antigen, and includes, for example, whole eggs, egg yolk and egg white of immunized eggs, these egg solutions and Solutions and egg extracts extracted with propanol and chloroform are included. It is preferable to include an egg yolk component from the viewpoint of the amount of the antibody contained. Powdered products such as spray-drying and freeze-drying methods are also included. Egg yolk is obtained by removing yolk lipid components from the yolk by a method using organic solvents such as hydroxypropyl methylcellulose phthalate, polyethylene glycol, dextran sulfate, propanol, ethanol, and hexane.
• organic solvents such as hydroxypropyl methylcellulose phthalate, polyethylene glycol, dextran sulfate, propanol, ethanol, and hexane.
• a whole egg which is spray-dried is referred to as a whole egg powder, and also includes a paste or liquid made from such a powder.
• the treated product includes the antibody itself purified from eggs by a known method such as ammonium sulfate salting out, sodium sulfate salting out, low-temperature ethanol precipitation, ion exchange chromatography, gel filtration, and affinity chromatography.
• an antibody obtained from an egg produced by a bird immunized with an antigen is used as a chicken.
• the total antibody titer is 256 times or more, preferably 25600 times or more, more preferably 51200 times or more, and still more preferably 1,638,400 to 13,107,200 times. It is measured based on the following neutralization test.
• CPE cytopathic effect
• Infection titer of parvovirus in the present invention was shown as 50% cell infectious dose (TCID 5 ). This calculation is based on the following method. Dilute the parvovirus serially 10-fold and add 5 wells per dilution to a 96-well plate for cell culture. Thereto was added an equal volume of suspension of FL74 cells are cultured in C0 2 incubator one 37 ° C. After 7 days, to confirm the presence or absence of CPE of each hole, TCID 5 from the dilution factor. calculate.
• Eggs and egg processed products produced by birds immunized with an antigen derived from parvovirus or parpovirus of the present invention have excellent preventive and therapeutic effects on parvovirus infection, and beagle dogs of 3 months old was administered at 2.0 g, preferably 1.5 g, and more preferably l.Og for 16 days per day as whole egg powder per animal, and parvovirus Y-1 strain was administered at 2 g / animal on the third day of administration.
• X 10 6 TCID 5 Characterized in that the beagle does not develop diarrhea when infected.
• the dose of the anti-parvovirus antibody in the above description is a dose when administered to a beagle dog having a standard weight of 3 months, that is, a beagle dog weighing about 2.5 to 3.5 kg.
• diarrhea has the meaning commonly used in the art, that is, a condition in which liquid or semisolid stool is abnormally repeatedly excreted from the intestine.
• Parvovirus Y-1 strain is available from the Ministry of Agriculture, Forestry and Fisheries Animal and Veterinary Laboratory.
• a 3-month-old beagle dog (body weight: 2.5 to 3.5 kg) is dissolved in a physiological saline solution as a whole egg powder with the egg or the processed egg product of the present invention, and three times a day (total egg powder 2.0 g / Daily, preferably 1.5 g / day, more preferably l. Og / day) daily by oral gavage.
• Test substance administration First day on day 3 1 hour after administration 10 6 TCID 5 .
• the antibody is administered for 14 days from that day, and the onset of diarrhea is observed.
• a control group a group to which no antibody is administered is established, and it is confirmed that diarrhea develops.
• the present invention also relates to an anti-parvovirus infection composition
• an anti-parvovirus infection composition is A composition that has the effect of ameliorating and treating symptoms such as diarrhea and vomiting in infected animals, and a composition that has the effect of preventing the infection of animals that have not been infected with parvovirus with parvovirus. I do.
• the anti-parvovirus infectious disease composition of the present invention contains the egg or the processed egg product of the present invention in an amount of usually 0.001 to 100% by mass, preferably 0.01 to 10% by mass, as the mass of the whole egg powder. Include so that it becomes a ratio.
• the egg or egg processed product of the present invention contains an anti-parvovirus antibody and has a high anti-parpovirus infectious disease activity, so that it works effectively even when the amount is relatively small, and the production cost and egg This is advantageous from the viewpoint of reducing the risk of allergy and calorie.
• an oral preparation such as a tablet, a granule, a powder, a capsule, a liquid, or the like, as it is or together with a conventional additive, is prepared by a usual preparation method.
• Additives include, for example, excipients, binders, disintegrants, lubricants, antioxidants, coloring agents, and flavoring agents, and are used as needed.
• it can be coated with a known retarder or the like.
• Excipients include, for example, sodium carboxymethylcellulose, agar, light caffeic anhydride, gelatin, crystalline cellulose, sorbitol, talc, dextrin, starch, lactose, sucrose, glucose, mannitol, magnesium aluminate metasilicate, phosphoric acid Hydrogen calcium or the like can be used.
• binders include gum arabic, sodium alginate, ethanol, ethylethylenolose, casein sodium, canolepoxymethylcellulose sodium, agar, purified water, gelatin, starch, tragacanth, lactose, and hydroxycellenol.
• Disintegrants include, for example, carboxymethylcellulose, sodium carboxymethinolecellulose, canolepox methoxymethinolese / rerose calcium, crystalline cenorellose, starch, hydroxypropyl starch and the like.
• examples of the lubricant include stearic acid, calcium stearate, magnesium stearate, talc, hydrogenated oil, sucrose fatty acid ester, and waxes.
• antioxidants examples include tocoprol, estenol gallate, dibutylhydroxytonolene (BHT), butynoledroxyanisonole (BHA), and ascorbic acid.
• BHT dibutylhydroxytonolene
• BHA butynoledroxyanisonole
• ascorbic acid examples include tocoprol, estenol gallate, dibutylhydroxytonolene (BHT), butynoledroxyanisonole (BHA), and ascorbic acid.
• Antacids sodium bicarbonate, magnesium carbonate, precipitated calcium carbonate, synthetic hydrotalcite, etc.
• gastric mucosal protective agents synthetic aluminum silicate, sucralfate, sodium copper chlorophyllin, etc.
• the dose of the anti-parvovirus infection composition of the present invention varies depending on conditions such as the animal to be administered, the condition of the animal, and the age of the animal. Is administered so as to be 0.005 to 70 g, preferably 0.005 to 7 g.
• the animal to which the egg or the processed egg product of the present invention and the anti-parvovirus infectious disease composition containing the same are given are not particularly limited as long as they can cause an infectious disease by parvovirus, and include, for example, mammals, Birds and the like. In the present invention, it is suitably used for mammals such as dogs, cats, humans, pests, stags, pomas, sheep, goats, and the like, and birds such as chickens and quail.
• the present invention relates to a pet food containing the egg or the processed egg product of the present invention.
• the treatment and prevention of parvovirus infection can be performed effectively by feeding the pets with the eggs or the processed food containing the eggs of the present invention. That is, the present invention also relates to a pet food for preventing and treating parvovirus infection.
• the eggs or egg processed product of the present invention as the mass of the whole egg powder, usually, from 0.001 to 100 weight 0/0, preferably a rate of 0.01 to 10 wt% Including.
• the pet food raw materials other than the eggs or processed eggs of the present invention include various components such as meat, seafood, vegetables, cereals, starch, oils and fats, fish meal, dietary fiber, dairy products, vitamins, minerals, and moisture. It is prepared and is in the form of raw, semi-living, dried, etc., but most of them are generally manufactured by mixing, stirring and molding the above raw materials.
• the pet food of the present invention can be produced by mixing the egg, the processed egg product or the anti-parvovirus antibody of the present invention at the stage of mixing the raw materials, or by adding and stirring at the final stage. . If heat treatment at a high temperature is included in the pet food manufacturing process, mix the egg, egg processed product or anti-parvovirus antibody of the present invention after the heat treatment to prevent inactivation of the anti-parvovirus antibody. Is preferred.
• the pet food of the present invention also includes pet cookies, biscuits, snacks, gums, nutrients, and the like. It can be produced by blending the egg, processed egg product or anti-parpovirus antibody of the present invention.
• the dosage of the pet food of the present invention varies depending on various conditions such as the animal to be administered, the condition of the animal, the age, etc., and is usually 0.005 per day of the animal's body weight per kg of the whole egg powder per day. 7070 g, preferably 0.005-7 g.
• the egg, egg processed product or pet food containing an anti-parvovirus antibody of the present invention is advantageous in terms of safety, palatability, and ease of administration. .
• the animal to which the egg, the processed egg or the pet food containing the anti-parvovirus antibody of the present invention is to be fed is not particularly limited as long as the animal can cause an infectious disease due to parvovirus, and examples thereof include mammals and birds. Is mentioned. It is suitably used for dogs, cats, araidama, pandas, mink, ferrets, weasels, wokami, scallops and foxes. In particular, pet food for dogs and cats is advantageous in terms of safety, palatability, and ease of administration.
• a probiotic agent can be added together with the egg, processed egg product or anti-parvovirus antibody of the present invention.
• symptoms such as vomiting and weight loss associated with diarrhea can be effectively reduced.
• probiotic live bacteria, yeasts and fungi, which can be manufactured by conventional methods in the art.
• the bacterium used as a probiotic agent preferably used in the present invention is not particularly limited, but includes, for example, lactic acid bacteria such as bacteria belonging to the genus Lactobacillus and Streptococcus, bacteria belonging to the genus Clostridium, and the genus Bifidobacterium. And other useful intestinal bacteria such as Enterococcus, Ratatococcus, and Pachilus.
• Yeasts as probiotic agents include those belonging to the genera Candida and Pichia.
• Fungi as probiotic agents are not particularly limited, and include, for example, fungi belonging to the genera Aspergillus and Saccharomyces.
• These probiotic agents may be used alone or in combination of two or more.
• a useful enteric bacterium as a probiotic agent, specifically, a bacterium belonging to the genus Bifidobacterium.
• the mixing ratio of the probiotic agent is not particularly limited, but usually, the ratio of 10 to 10 11 probiotic agents to whole egg powder lg, preferably Is used at a ratio of 10 4 to 10 9 viable bacterial agents to the whole egg powder lg.
• This description includes part or all of the contents as disclosed in the description of Japanese Patent Application No. 2003-1504825, which is a priority document of the present application.
• the canine parvovirus CPK-2 strain was cultured in cat kidney cells (CRFK), and virus particles were collected by ultracentrifugation. This virus, 10 9 TCID 5 .
• the first immunization was carried out by emulsifying a solution containing the compound and Freund's adjuvant and injecting lml into the left and right pectoral muscles of a 12-week-old hen.
• a second immunization was performed 6 weeks later. Two weeks after the second immunization, the titer of the eggs laid by the chicken as measured by the neutralization reaction of the anti-Palpovirus antibody was 1,638,400-fold. This antibody titer lasted for 4 months.
• the eggs were collected and spray-dried to produce whole egg powder.
• the whole egg powder was diluted 8-fold, and the supernatant extracted with chloroform was used as a sample for neutralizing antibody titer measurement. In addition, this 8 times was also considered in the highest dilution factor at the time of judgment.
• This sample was serially diluted two-fold to an appropriate multiple with the diluent. That in each dilution, the infectivity titer 2 00 TCID 5.
• An equal volume of the virus antigen adjusted to / mL was added and mixed well. After placing this mixture at 37 ° C for 1 hour, 4 wells were added per dilution to a 96-well plate for cell culture.
• the canine parvovirus strain Cp83016 was cultured in cat kidney cells (CRFK), and virus particles were collected by ultracentrifugation. This virus 10 9 TCID 5 . And a Freund's adjuvant were subjected to a first immunization by injecting lml each into the left and right pectoral muscles of a 12-week-old hen. Similarly, a second immunization was performed 6 weeks later. Two weeks after the second immunization, the eggs laid by the chickens were collected and spray-dried to produce whole egg powder. When the neutralizing antibody titer was measured in the same manner as in Example 1, the neutralizing antibody titer of this whole egg powder was 51200 times.
• the canine parpovirus CPK-2 strain was cultured in cat kidney cells (CRFK), and virus particles were collected by ultracentrifugation. This virus 10 9 TCID 5 .
• the first immunization was carried out by emulsifying a solution containing the compound and Freund's adjuvant and injecting lml into the left and right pectoral muscles of a 12-week-old hen.
• a second immunization was performed 6 weeks later.
• the neutralizing antibody titer measured by the neutralization reaction of the anti-parvovirus antibody in the yolk fluid of the eggs laid by the chicken was 1,638,400-fold. This antibody titer lasted for 4 months.
• the eggs were collected and spray-dried to make a whole egg powder.
• the neutralizing antibody titer of this powder was 1,638,400-fold.
• the eggs and processed egg products of the present invention specifically act on parvovirus pathogens, so they exhibit an extremely superior preventive effect on animal parvovirus infection compared to vaccines, and also provide parvovirus infection It can reduce symptoms such as diarrhea, vomiting, and weight loss, which are specific to the disease. Furthermore, the eggs and processed eggs of the present invention have a very high neutralizing antibody titer against parvovirus and can prevent diarrhea caused by parvovirus infection very effectively. It can be used as a preventive drug, a therapeutic drug and an animal food with an extremely excellent anti-parvovirus infection ability.

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Citations (4)

• 2003
  • 2003-05-30 JP JP2003154825A patent/JP2004352693A/ja active Pending
• 2004
  • 2004-05-20 WO PCT/JP2004/007232 patent/WO2004105792A1/ja active Application Filing
  • 2004-05-28 TW TW093115407A patent/TW200509963A/zh unknown

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