CN112293576A - Application of chlorogenic acid in preparation of medicine or feed additive for preventing and/or treating canine parvovirus disease and canine distemper - Google Patents
Application of chlorogenic acid in preparation of medicine or feed additive for preventing and/or treating canine parvovirus disease and canine distemper Download PDFInfo
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- CN112293576A CN112293576A CN202011192544.4A CN202011192544A CN112293576A CN 112293576 A CN112293576 A CN 112293576A CN 202011192544 A CN202011192544 A CN 202011192544A CN 112293576 A CN112293576 A CN 112293576A
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- PZIRUHCJZBGLDY-UHFFFAOYSA-N Caffeoylquinic acid Natural products CC(CCC(=O)C(C)C1C(=O)CC2C3CC(O)C4CC(O)CCC4(C)C3CCC12C)C(=O)O PZIRUHCJZBGLDY-UHFFFAOYSA-N 0.000 title claims abstract description 62
- CWVRJTMFETXNAD-KLZCAUPSSA-N Neochlorogenin-saeure Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-KLZCAUPSSA-N 0.000 title claims abstract description 62
- CWVRJTMFETXNAD-JUHZACGLSA-N chlorogenic acid Chemical compound O[C@@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-JUHZACGLSA-N 0.000 title claims abstract description 62
- 229940074393 chlorogenic acid Drugs 0.000 title claims abstract description 62
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Classifications
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- A—HUMAN NECESSITIES
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- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/111—Aromatic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/40—Feeding-stuffs specially adapted for particular animals for carnivorous animals, e.g. cats or dogs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/216—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acids having aromatic rings, e.g. benactizyne, clofibrate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
- A61K36/355—Lonicera (honeysuckle)
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/46—Eucommiaceae (Eucommia family), e.g. hardy rubber tree
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Natural Medicines & Medicinal Plants (AREA)
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- Veterinary Medicine (AREA)
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- Animal Behavior & Ethology (AREA)
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Abstract
The invention discloses application of chlorogenic acid in preparing a medicament or feed additive for preventing and/or treating canine parvovirus disease and canine distemper. Experimental results show that after the medicine prepared by taking chlorogenic acid as the only active ingredient is applied, the body temperature, spirit, appetite and excrement state of the puppy suffering from the canine parvovirus disease are obviously improved, the cure rate of the medicine on the canine parvovirus disease is 67%, the significant efficiency is 33%, and the total effective rate is 100%; after the medicine prepared by taking chlorogenic acid as the only active ingredient is applied, the body temperature, spirit, appetite and excrement state of dogs suffering from canine distemper are obviously improved, the cure rate of the medicine on the canine distemper is 58%, the obvious effect rate is 25%, and the total effective rate is 83%. Chlorogenic acid as the only active ingredient has good application prospect in preparing medicines or feed additives for preventing and/or treating parvovirus diseases and canine distemper.
Description
Technical Field
The invention belongs to the field of pharmacy, and particularly relates to application of chlorogenic acid serving as a unique active ingredient in preparation of a medicine or feed additive for preventing and/or treating canine parvovirus disease and canine distemper.
Background
With the vigorous development of the pet industry, the number of dog breeders has been increasing dramatically. In recent years, canine parvovirus disease and the abuse of canine distemper have caused great harm to the canine industry and canine breeders. The high morbidity and mortality rate makes canine parvovirus disease and canine distemper terrible diseases threatening the health of canines.
Canine Parvovirus (CPV) belongs to the family of Parvoviridae (Parvoviridae), the subfamily of parvoviruses (Parvovirinae), the genus Protoparvovirus (Protoparvovirus), and includes other viruses such as Feline Parvovirus (FPV) and Mink Enteritis Virus (MEV). Canine Parvovirus disease (CP) is a virulent infectious disease caused by Canine Parvovirus, and has the greatest harm to 2-6 month-old puppies, and vomiting, diarrhea and hemorrhagic enteritis are main clinical manifestations.
Canine Distemper (CD) is a highly contagious disease caused by Canine Distemper Viruses (CDV), and has strong infectivity and a death rate of over 80 percent. The body temperature of the dogs at the early stage of canine distemper symptom is as high as 39.5-41 ℃, the dogs have inappetence, depression, watery secretion flowing out of eyes and nose, sneezing and diarrhea. The body temperature rise, cough, purulent nasal discharge and purulent eye droppings occur again within 2-14 days later, and meanwhile, the secondary gastrointestinal diseases, vomiting, diarrhea and inappetence are caused. Mental depression and lethargy. Typical neurological symptoms, white foam and convulsion, appear in the later period of canine distemper, and the death rate is as high as 80%.
At present, no particularly effective medicine is available for preventing and treating canine parvovirus diseases and canine distemper, immunization is mainly considered, and generally inactivated vaccines and attenuated vaccines with lower production cost are used. The main anti-canine parvovirus vaccine used in China is CPV-2b type, but the vaccine is proved to have no complete immune effect on puppies; the vaccine for resisting canine distemper virus is a CDV-11 strain or a combined vaccine, can only play a certain role in preventing canine distemper and cannot be used for treating the canine distemper virus.
Therefore, there is a high necessity to develop a drug capable of effectively treating canine parvovirus disease and canine distemper. At present, no report on the use of chlorogenic acid as the only active ingredient in the preparation of medicaments for preventing and/or treating canine parvovirus disease and canine distemper exists.
Disclosure of Invention
The invention aims to provide application of chlorogenic acid serving as a unique active ingredient in preparation of medicines or feed additives for preventing and/or treating canine parvovirus diseases and canine distemper.
The invention provides application of chlorogenic acid as a unique active ingredient in preparing a medicament or feed additive for preventing and/or treating parvovirus diseases and/or distemper.
The invention also provides application of the traditional Chinese medicine extract in preparing a medicine or feed additive for preventing and/or treating parvovirus diseases and/or distemper diseases, wherein the content of chlorogenic acid in the traditional Chinese medicine extract is 20.00-100.00 wt.%.
Further, the content of chlorogenic acid in the traditional Chinese medicine extract is 21.36 wt% -98.00 wt%.
Further, the traditional Chinese medicine extract is an eucommia ulmoides leaf extract or a honeysuckle extract.
Further, the preparation method of the eucommia ulmoides leaf extract or the honeysuckle extract comprises the following steps: decocting folium Eucommiae or flos Lonicerae in water, filtering, concentrating the filtrate, and drying.
Further, the mass ratio of the eucommia ulmoides leaves or the honeysuckle to water is 1: 10-1: 15; the decoction time is 0.5-1.0 hour, and the decoction times are 2-3 times.
Further, after the concentration, the method also comprises a purification step, wherein the purification method is one or more than two of precipitation, extraction, chromatographic column adsorption and desorption and crystallization;
preferably, the purification method comprises the steps of precipitation, extraction and crystallization in sequence, or the steps of precipitation, chromatographic column adsorption and desorption and crystallization in sequence.
Further, a reagent adopted by the precipitation is ethanol, a reagent adopted by the extraction is ethyl acetate, a reagent adopted by the crystallization is ethyl acetate, and macroporous adsorption resin adopted by the chromatographic column is HPD-100 or D-101.
Further, the parvovirus disease is canine parvovirus disease or chicken parvovirus disease, preferably canine parvovirus disease;
or the distemper is canine distemper.
Further, each unit preparation of the medicine or the feed additive contains 10mg-100mg of chlorogenic acid;
and/or the medicine or feed additive is an injection or a solid preparation.
Experimental results show that after the medicine prepared by taking chlorogenic acid as the only active ingredient is applied, the body temperature, spirit, appetite and excrement state of the puppy suffering from the canine parvovirus disease are obviously improved, the cure rate of the medicine on the canine parvovirus disease is 67%, the significant efficiency is 33%, and the total effective rate is 100%; after the medicine prepared by taking chlorogenic acid as the only active ingredient is applied, the body temperature, spirit, appetite and excrement state of dogs suffering from canine distemper are obviously improved, the cure rate of the medicine on the canine distemper is 58%, the obvious effect rate is 25%, and the total effective rate is 83%. The chlorogenic acid has a remarkable effect of treating canine parvovirus diseases and canine distemper, can be used for treating canine parvovirus diseases and canine distemper by being used alone, and is high in effective rate. Chlorogenic acid as the only active ingredient has good application prospect in preparing medicines or feed additives for preventing and/or treating canine parvovirus disease and canine distemper.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Detailed Description
The raw materials and equipment used in the invention are known products and are obtained by purchasing commercial products.
Example one preparation method of folium Eucommiae extract of the present invention
The medicinal materials are as follows: eucommia ulmoides leaf with chlorogenic acid content of 3.58 wt.%.
The preparation method comprises the following steps: decocting folium cortex eucommiae with 10 times of water for two times, each time for 0.5 hour, filtering decoction, combining filtrates, concentrating to 1/10 of the volume of a stock solution, stirring, adding 5 times of ethanol of the volume of a concentrated solution, standing, filtering, concentrating the filtrate, recovering the ethanol, and performing spray drying to obtain the folium cortex eucommiae extract, wherein the content of chlorogenic acid is 21.36-26.75 wt.%.
Example II preparation of extract of eucommia ulmoides Oliver of the present invention
The medicinal materials are as follows: eucommia ulmoides leaf with chlorogenic acid content of 3.58 wt.%.
The preparation method comprises the following steps: decocting folium cortex eucommiae with 10 times of water for two times, each time for 0.5 hour, filtering decoction, combining filtrates, concentrating to 1/10 of the volume of a stock solution, stirring, adding 5 times of ethanol of the volume of a concentrated solution, standing, filtering, concentrating the filtrate under reduced pressure, recovering ethanol, concentrating to a relative density of 1.05-1.15(25 ℃), extracting for 5 times with 15 times of ethyl acetate of the volume of the concentrated solution, retaining an ethyl acetate phase, concentrating, and volatilizing at low temperature to obtain an folium cortex eucommiae extract, wherein the content of chlorogenic acid is 55.81-63.47 wt.%.
Example III preparation of eucommia ulmoides leaf extract of the present invention
The medicinal materials are as follows: eucommia ulmoides leaf with chlorogenic acid content of 3.58 wt.%.
The preparation method comprises the following steps: decocting folium Eucommiae in 10 times of water for 0.5 hr twice, filtering the decoctions, mixing the filtrates, concentrating to 1/10 volume of the stock solution, stirring, adding 5 times of ethanol, standing, filtering, concentrating the filtrate under reduced pressure, recovering ethanol, concentrating to relative density of 1.05-1.15(25 deg.C), extracting with 15 times of ethyl acetate for 5 times, retaining ethyl acetate phase, concentrating, standing at 0-10 deg.C for 12 hr for crystallization, filtering, and drying the crystal at low temperature to obtain folium Eucommiae extract with chlorogenic acid content of 80.24-86.09 wt.%.
Example four preparation of extract of eucommia ulmoides leaves of the present invention
The medicinal materials are as follows: eucommia ulmoides leaf with chlorogenic acid content of 3.58 wt.%.
The preparation method comprises the following steps: decocting folium Eucommiae with 10 times of water for 0.5 hr twice, filtering the decoctions, mixing the filtrates, concentrating to 1/10 volume of the stock solution, stirring, adding 5 times of ethanol, standing, filtering, concentrating the filtrate under reduced pressure, recovering ethanol, concentrating to relative density of 1.05-1.15(25 deg.C), adding 15 times of ethyl acetate, extracting for 5 times, retaining ethyl acetate phase, concentrating, standing at 0-10 deg.C for 12 hr for crystallization, filtering, dissolving with ethyl acetate at 70 deg.C, standing at 0-10 deg.C for 12 hr for recrystallization, filtering, and drying at low temperature to obtain folium Eucommiae extract with chlorogenic acid content of not less than 98 wt.%.
Example five preparation methods of honeysuckle extract of the invention
The medicinal materials are as follows: honeysuckle flower, chlorogenic acid content is 2.88%.
The preparation method comprises the following steps: taking honeysuckle, adding 15 times of water by weight, decocting for three times, each time for 0.5 hour, filtering decoction, combining filtrates, concentrating to 1/10 of the volume of a stock solution, stirring, adding 8 times of ethanol of the volume of a concentrated solution, standing, filtering, concentrating the filtrate, recovering the ethanol, and performing spray drying to obtain the honeysuckle extract, wherein the content of chlorogenic acid is 15.78-20.52 wt.%.
EXAMPLE six preparation method of honeysuckle extract of the present invention
The medicinal materials are as follows: honeysuckle flower, chlorogenic acid content is 2.88%.
The preparation method comprises the following steps: taking honeysuckle, adding 15 times of water for decocting for three times, each time for 0.5 hour, filtering decoction, combining filtrates, concentrating to 1/10 of the volume of a stock solution, stirring, adding 8 times of ethanol of the volume of a concentrated solution, standing, filtering, concentrating the filtrate, recovering ethanol, adsorbing by using an HPL-100 macroporous adsorption resin chromatographic column, eluting by using 60% ethanol, collecting eluent, concentrating, recovering ethanol, and performing spray drying to obtain the honeysuckle extract, wherein the content of chlorogenic acid is 43.21-48.77 wt.%.
EXAMPLE seven preparation method of honeysuckle extract of the invention
The medicinal materials are as follows: honeysuckle flower, chlorogenic acid content is 2.88%.
The preparation method comprises the following steps: decocting honeysuckle flower with 15 times of water for three times, each time for 0.5 hour, filtering decoction, combining filtrates, concentrating to 1/10 of the volume of a stock solution, stirring, adding 8 times of ethanol of the volume of a concentrated solution, standing, filtering, concentrating the filtrate, recovering ethanol, adsorbing by using an HPL-100 macroporous adsorption resin chromatographic column, eluting with 60% ethanol, collecting eluate, concentrating, recovering ethanol, adsorbing the concentrated solution by using a D-101 macroporous adsorption resin chromatographic column, sequentially and gradiently eluting with 10%, 30% and 50% ethanol, collecting 50% eluate, concentrating, recovering ethanol, and spray drying to obtain the honeysuckle flower extract, wherein the content of chlorogenic acid is 72.35-80.13 wt.%.
EXAMPLE VIII preparation of the honeysuckle extract of the present invention
The medicinal materials are as follows: honeysuckle flower, chlorogenic acid content is 2.88%.
The preparation method comprises the following steps: decocting flos Lonicerae with 15 times of water for three times, each for 0.5 hr, filtering the decoction, mixing the filtrates, concentrating to 1/10 volume of the stock solution, stirring, adding 8 times of ethanol, standing, filtering, concentrating the filtrate, recovering ethanol, adsorbing with HPL-100 macroporous adsorbent resin chromatographic column, eluting with 60% ethanol, collecting eluate, concentrating, recovering ethanol, adsorbing the concentrate with D-101 macroporous adsorbent resin chromatographic column, gradually eluting with 10%, 30% and 50% ethanol, collecting 50% eluate, concentrating, recovering ethanol, concentrating to obtain extract, dissolving with ethyl acetate at 60 deg.C, cooling to room temperature, crystallizing, filtering, and drying to obtain flos Lonicerae extract with chlorogenic acid content of not less than 95 wt.%.
Example nine preparation methods of the drug powder injection of the invention
Prescription one
The preparation method comprises the following steps: weighing folium Eucommiae extract and glucose according to the prescription, dissolving with injectable water, filtering, spray drying, aseptically packaging into 1000 bottles.
EXAMPLE Ten preparation methods of the pharmaceutical powder injection of the present invention
Prescription two
The preparation method comprises the following steps: weighing folium Eucommiae extract, glucose and sodium chloride according to the prescription, dissolving with injectable water, filtering, spray drying, aseptically packaging into 1000 bottles.
EXAMPLE eleven preparation method III of the pharmaceutical powder injection of the present invention
Prescription three
The preparation method comprises the following steps: weighing folium Eucommiae extract and mannitol according to the prescription, dissolving in injectable water, filtering, spray drying, aseptically packaging into 1000 bottles.
Example twelve preparation methods of the drug powder injection of the invention
Prescription four
The preparation method comprises the following steps: weighing folium Eucommiae extract, and packaging into 1000 bottles.
Thirteen embodiment preparation method five of the drug powder injection
Prescription five
The preparation method comprises the following steps: weighing folium Eucommiae extract, glucose, and sodium bisulfite according to the prescription, dissolving with injectable water, filtering, and bottling into 1000 bottles.
Example fourteen preparation methods of the drug powder injection of the invention
Prescription 6
The preparation method comprises the following steps: weighing folium Eucommiae extract and vitamin C according to the prescription, dissolving in injectable water, filtering, and bottling into 1000 bottles.
Fifteen process for preparing the medicament powder for injection of the invention
Prescription 7
The preparation method comprises the following steps: weighing flos Lonicerae extract, glucose, and sodium bisulfite according to the prescription, dissolving with injectable water, filtering, and bottling into 1000 bottles.
The beneficial effects of the present invention are demonstrated by the following experimental examples.
Experimental example 1 curative Effect of chlorogenic acid on Canine parvovirus disease
1. Test materials
1.1 medicaments
Chlorogenic acid, 99.2 wt.% in content, was provided by the scientific and technological limited, chapter nine, Sichuan. Before use, the chlorogenic acid is dissolved by normal saline to prepare about 15mg per 1 mL.
Physiological saline was manufactured by Sichuan Koran pharmaceutical Co.
1.2 animals
12 Beagle dogs are used as both male and female (female is not pregnant), are 5-6 months old and have 6.0-7.0 kg of body weight, and are provided by dog farms of laboratory animal institute of medical academy of sciences of Sichuan province.
1.3 viruses
Canine parvovirus, provided by Qingdao animal and plant quarantine office.
2. Test method
2.1 model building
After 6 days, the dog parvovirus challenge is carried out on healthy and non-immunized dogs, and the clinical symptoms of the typical dog parvovirus disease appear: the body temperature is raised to 40.1-41.0 ℃, the mental depression, the loss of appetite, the vomiting, the diarrhea, the bloody stool and the like are detected by adopting a hemagglutination test, a hemagglutination inhibition test and a canine parvovirus rapid diagnosis test paper respectively, and the canine parvovirus disease is confirmed to be suffered.
2.2 animal administration
12 Beagle dogs suffering from canine parvovirus disease are randomly divided into 2 groups, and 6 dogs in each group are respectively a chlorogenic acid medicament treatment group and a negative control group.
The chlorogenic acid drug treatment group is injected with chlorogenic acid physiological saline solution at 2mg/kg muscle once a day for 10 days; the negative control group was injected intramuscularly with the same volume of saline.
2.3 clinical symptom Observation
Body temperature was measured 1 time in the morning and evening each day, and dog's defecation, mental status, food intake and water intake were observed and recorded. The clinical symptoms, the disease course progression and the treatment effect of the sick dogs in each group are judged and compared. Analyzing and collating the results, classifying the healed and effective patients as effective cases, and counting the effective rate.
2.4 criteria for therapeutic efficacy
(1) The clinical symptoms after the healing use are as follows: body temperature, spirit and appetite return to normal, bloody stool stops, stool forms, urine is normal and the like.
(2) The clinical symptoms after the effective use are as follows: the body temperature, the spirit and the appetite return to normal, but the feces are not formed or the body temperature and the spirit return to normal, but the appetite is poor, the feces do not form, and the like.
(3) The clinical symptoms are not improved or worsened and die after the ineffective medication.
3. Test results
TABLE 1 Observation of clinical symptoms at day 10 after dosing
Note: "/" indicates death.
TABLE 2 curative effective rate of chlorogenic acid drug treatment group
Compared with the negative control group, Beagle dogs with canine parvovirus disease have obvious improvement on body temperature, spirit, appetite and fecal state after intramuscular injection of chlorogenic acid solution.
In the treatment of Beagle dogs suffering from the artificial hair dye canine parvovirus disease, through the treatment statistics of administering chlorogenic acid through intramuscular injection for 10 days, 4 dogs are healed, 2 dogs are effective, none of the animals die, the healing rate is 67%, the significant efficiency is 33%, and the total effective rate is 100%, so that the injection taking the chlorogenic acid as the only active ingredient has good clinical application prospect in treating the canine parvovirus disease.
Experimental example 2 curative effect test of chlorogenic acid on canine distemper
1. Test materials
1.1 medicaments
Chlorogenic acid, 99.2 wt.% in content, was provided by the scientific and technological limited, chapter nine, Sichuan. Before use, the chlorogenic acid is dissolved by normal saline to prepare about 15mg per 1 mL.
Physiological saline was supplied by Sichuan Konlun pharmaceutical Co.
Canine pentaserum, provided by the military veterinary institute of military medical academy of sciences.
1.2 animals
36 sick dogs with confirmed canine distemper provided by a pet hospital in Kyowa west are male and female half, the age is 1-2 years, and the weight is 12-14 kg,
2. test method
2.1 model building
36 diagnosed dogs were randomized into 3 groups of 12 dogs each: and observing and recording physiological indexes and clinical performances of the sick dogs in a positive drug group, a chlorogenic acid drug treatment group and a negative control group.
2.2 animal administration
Dosing was performed according to experimental groups:
(1) positive drug group: injecting quintuplet serum into muscle, 1 ml/kg/time, once daily, and continuously administering for 15 days;
(2) chlorogenic acid drug group: intramuscular injection of chlorogenic acid in normal saline solution at a dose of 2 mg/kg/time, once a day, for 15 days;
(3) negative control group: the same volume of physiological saline as that of the group (2) was injected intramuscularly once a day for 15 days.
2.3 clinical symptom Observation
Body temperature was measured 1 time in the morning and evening each day, and dog's defecation, mental status, food intake and water intake were observed and recorded. The clinical symptoms, the disease course progression and the treatment effect of the sick dogs in each group are judged and compared. Analyzing and collating the results, classifying the healed and effective patients as effective cases, and counting the effective rate.
2.4 criteria for therapeutic efficacy
(1) The clinical symptoms after the healing use are as follows: the body temperature of the sick dog is reduced to 38.5 ℃ or below, clinical symptoms such as listlessness, diarrhea and the like do not appear any more, and the appetite is recovered to be normal.
(2) The clinical symptoms after the effective use are as follows: body temperature, spirit, appetite and 2 or more items in the stool are normal.
(3) Clinical symptoms after ineffective medication: those who do not improve or worsen, die.
3. Test results
TABLE 3 Observation of clinical symptoms at day 15 after dosing
Note: "/" indicates death.
TABLE 4 therapeutic Effect of the respective administration groups
Compared with a negative control group, the positive drug and the chlorogenic acid can obviously improve the body temperature, spirit, appetite and fecal state of a sick dog suffering from canine distemper.
From the table above, in the treatment of the canine distemper, 3 dogs are cured, 4 dogs are effective, 5 dogs are ineffective, the cure rate is 25%, the effective rate is 33%, and the total effective rate is 58% by 15-day treatment of the positive medicament through intramuscular injection; counting the treatment effect of chlorogenic acid for 15 days, wherein 7 patients are cured, 3 patients are effective, 2 patients are ineffective, the cure rate is 58%, the effective rate is 25%, and the total effective rate is 83%; none of the negative controls were effective. Compared with a negative control group, the positive medicine and the chlorogenic acid obviously improve the body temperature, the spirit, the appetite and the excrement state of a sick dog suffering from canine distemper; the injection taking chlorogenic acid as the only active ingredient has better treatment effect on canine distemper compared with the quintuplet serum which is a positive medicament, and has good clinical application prospect.
In conclusion, the invention provides the application of chlorogenic acid as a unique active ingredient in preparing a medicament or a feed additive for preventing and/or treating canine parvovirus disease and canine distemper. Experimental results show that after the medicine prepared by taking chlorogenic acid as the only active ingredient is applied, the body temperature, spirit, appetite and excrement state of the puppy suffering from the canine parvovirus disease are obviously improved, the cure rate of the medicine on the canine parvovirus disease is 67%, the significant efficiency is 33%, and the total effective rate is 100%; after the medicine prepared by taking chlorogenic acid as the only active ingredient is applied, the body temperature, spirit, appetite and excrement state of dogs suffering from canine distemper are obviously improved, the cure rate of the medicine on the canine distemper is 58%, the obvious effect rate is 25%, and the total effective rate is 83%. The chlorogenic acid has a remarkable effect of treating canine parvovirus diseases and canine distemper, can be used for treating canine parvovirus diseases and canine distemper by being used alone, and is high in effective rate. Chlorogenic acid as the only active ingredient has good application prospect in preparing medicines or feed additives for preventing and/or treating canine parvovirus disease and canine distemper.
Claims (10)
1. Application of chlorogenic acid as a sole active ingredient in preparing a medicament or feed additive for preventing and/or treating parvovirus diseases and/or distemper diseases.
2. Use of a traditional Chinese medicine extract in preparation of a medicament or feed additive for preventing and/or treating parvovirus diseases and/or distemper diseases, wherein the content of chlorogenic acid in the traditional Chinese medicine extract is 20.00-100.00 wt.%.
3. Use according to claim 2, characterized in that: the content of chlorogenic acid in the Chinese medicinal extract is 21.36-98.00 wt.%.
4. Use according to claim 2 or 3, characterized in that: the Chinese medicinal extract is folium Eucommiae extract or flos Lonicerae extract.
5. Use according to claim 4, characterized in that: the preparation method of the eucommia ulmoides leaf extract or the honeysuckle extract comprises the following steps: decocting folium Eucommiae or flos Lonicerae in water, filtering, concentrating the filtrate, and drying.
6. Use according to claim 5, characterized in that: the mass ratio of the eucommia ulmoides leaves or the honeysuckle to water is 1: 10-1: 15; the decoction time is 0.5-1.0 hour, and the decoction times are 2-3 times.
7. Use according to claim 6, characterized in that: after the concentration, the method also comprises a purification step, wherein the purification method is one or more than two of precipitation, extraction, chromatographic column adsorption and desorption and crystallization;
preferably, the purification method comprises the steps of precipitation, extraction and crystallization in sequence, or the steps of precipitation, chromatographic column adsorption and desorption and crystallization in sequence.
8. Use according to claim 7, characterized in that: the reagent adopted by the precipitation is ethanol, the reagent adopted by the extraction is ethyl acetate, the reagent adopted by the crystallization is ethyl acetate, and the macroporous adsorption resin adopted by the chromatographic column is HPD-100 or D-101.
9. Use according to any one of claims 1 to 8, characterized in that: the parvovirus disease is canine parvovirus disease or chicken parvovirus disease, preferably canine parvovirus disease;
or the distemper is canine distemper.
10. Use according to any one of claims 1 to 8, characterized in that: each unit preparation of the medicine or the feed additive contains 10mg-100mg of chlorogenic acid;
and/or the medicine or feed additive is an injection or a solid preparation.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102512547A (en) * | 2011-12-14 | 2012-06-27 | 赵兴友 | Traditional Chinese medicine composition for treating canine distemper virus and preparation method thereof |
| CN104873634A (en) * | 2015-06-11 | 2015-09-02 | 刘洋 | Traditional Chinese medicine type dog food additive for preventing canine distemper of puppies as well as preparation method and use method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102512547A (en) * | 2011-12-14 | 2012-06-27 | 赵兴友 | Traditional Chinese medicine composition for treating canine distemper virus and preparation method thereof |
| CN104873634A (en) * | 2015-06-11 | 2015-09-02 | 刘洋 | Traditional Chinese medicine type dog food additive for preventing canine distemper of puppies as well as preparation method and use method thereof |
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| 杨岸奇;李朝晖;张善文;: "异绿原酸在畜牧生产的应用前景", 湖南饲料 * |
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