KR20230092754A - Composition for treating infection of feline infectious peritonitis comprising chlortetracycline - Google Patents

Abstract

The present invention relates to a composition for the treatment of feline infectious peritonitis virus (FIPV) infection containing chlortetracycline, and more particularly, to a composition for preventing or treating FIPV infection containing chlortetracycline, which has a very excellent ability to inhibit FIPV enzyme activity and plaque formation. It relates to a veterinary composition, a feed additive, a health food composition, and a method for treating FIPV infection by administering the same.
The composition of the present invention comprising chlortetracycline or a veterinarily acceptable salt thereof inhibits the enzymatic activity of feline infectious peritonitis virus and has a very high activity to inhibit the virus from proliferating intracellularly and forming plaques, thereby preventing feline infectious disease. It can be very useful for preventing peritonitis or developing a therapeutic agent.

Description

Composition for treating feline infectious peritonitis comprising chlortetracycline {Composition for treating infection of feline infectious peritonitis comprising chlortetracycline}

The present invention relates to a composition for the treatment of feline infectious peritonitis virus (FIPV) infection containing chlortetracycline, and more particularly, to a composition for preventing or treating FIPV infection containing chlortetracycline, which has a very excellent ability to inhibit FIPV enzyme activity and plaque formation. It relates to a veterinary composition, a feed additive, a health food composition, and a method for treating FIPV infection by administering the same.

Feline infectious peritonitis occurs mainly in young feline animals before one year of age, and the induction rate is about 17%, but the fatality rate reaches almost 100%, making it the most fatal and difficult to treat cat disease.

Feline infectious peritonitis (FIP) is caused by Feline Infectious Peritonitis Virus (FIPV), which binds strongly to cells and tissues, causing lesions in the collecting duct or barrier of the kidneys and contaminated food or bedding. , characterized by transmission through urine or feces. Infected individuals are initially asymptomatic for a considerable period of time and act as disease vectors, resulting in a high risk of transmission that can spread the virus for a long period of time. At the time of onset, clinical symptoms such as anorexia, fever, jaundice, diarrhea, weight loss, neurological symptoms, and uveitis are displayed, and ascites gradually fills up and dyspnea occurs. It's hard. In addition, it is a very fatal disease with a very high mortality rate and no proper treatment method is known once it develops. The effectiveness of preventive vaccines is still controversial.

Currently, there are no approved vaccines or effective antiviral therapies for the treatment of FIP. Thus, there is a significant need for the development of compounds to treat FIP in cats.

Accordingly, as a result of intensive research to discover substances capable of preventing or treating FIP, the inventors of the present invention found that chlortetracycline exhibits excellent FIPV enzyme activity inhibitory ability and plaque formation inhibitory ability, and completed the present invention. .

Therefore, an object of the present invention is to provide a veterinary composition, animal feed additive, or health food composition for preventing or treating feline infectious peritonitis (FIP), which includes chlortetracycline or a veterinarily acceptable salt thereof as an active ingredient. is to provide

Another object of the present invention is to provide a method for treating feline infectious peritonitis, comprising administering chlortetracycline or a veterinarily acceptable salt thereof to a non-human mammal.

Another object of the present invention is to provide an antiviral composition against feline infectious peritonitis virus comprising chlortetracycline or a veterinarily acceptable salt thereof.

In order to achieve the above object of the present invention, the present invention provides a veterinary composition for preventing or treating feline infectious peritonitis (FIP) comprising chlortetracycline or a veterinarily acceptable salt thereof as an active ingredient, animals It provides a feed additive or health food composition.

In order to achieve another object of the present invention, the present invention provides a method for treating feline infectious peritonitis, comprising administering chlortetracycline or a veterinarily acceptable salt thereof to a non-human mammal.

In order to achieve another object of the present invention, the present invention provides an antiviral composition against feline infectious peritonitis virus comprising chlortetracycline or a veterinarily acceptable salt thereof.

Hereinafter, the present invention will be described in detail.

The present invention provides a veterinary composition for preventing or treating feline infectious peritonitis (FIP) comprising chlortetracycline or a veterinarily acceptable salt thereof as an active ingredient.

Chlortetracycline, a substance used for the treatment of respiratory diseases, is a tetracycline-based antibacterial agent having the structure of Formula 1 below, and is a broad-spectrum antibacterial agent that acts on Gram-negative, Gram-positive, chlamydia, rickettsiae, and mycoplasma. Chlortetracycline acts on the bacterial 30S ribosome to block aminoacyl-tRNA binding and consequently inhibits protein synthesis, thereby inhibiting bacterial growth. Neomycin is an aminoglycoside-based antibacterial agent that binds to the ribosome 30S subunit of bacteria and blocks protein synthesis. am. Loperamide is a piperidine derivative, similar in chemical structure to pethidine, an opioid, and exhibits antidiarrheal action by inhibiting intestinal peristalsis.

[Formula 1]

The composition of the present invention is characterized by enhancing resistance to FIPV or inhibiting the activity and proliferation of FIPV so as to reduce the risk of infection by FIPV and reduce the induction rate even when in contact with the virus.

As used herein, the term "treatment" or "treating" refers to the application or administration of a therapeutic agent, that is, the present chlortetracycline (alone or in combination with other veterinary agents) to a mammal, or feline infectious peritonitis, cat Treatment, cure, alleviation, alleviation, alteration, resolution, amelioration, enhancement of, or feline infectious peritonitis, feline infectious peritonitis in a mammal at risk of developing feline infectious peritonitis or symptoms of infectious peritonitis (e.g., diagnosis or ex vivo application) It is defined as the application or administration of a therapeutic agent to a mammal, isolated tissue or cell line for the purpose of influencing the symptoms of or the likelihood of developing feline infectious peritonitis. Such treatment can be tailored and modified based on knowledge obtained, inter alia, from the field of genomic pharmacology.

As used herein, the term "prevent" or "prevention" means that, if nothing has happened, there is no development of the disorder or disease, or if there has already been the development of the disorder or disease, there is no further development of the disorder or disease. Also contemplated is one's ability to prevent some or all of the symptoms associated with a disorder or disease.

As used herein, the terms "mammal", "individual" or "subject" refer to mammals other than humans. For example, non-human mammals include livestock and pets such as ovine, bovine, porcine, canine, feline and murine mammals. Preferably the mammal, subject or individual may be a cat.

As used herein, the terms "effective amount", "veterinarily effective amount" and "therapeutically effective amount" refer to a non-toxic sufficient amount of an agent to provide a desired biological result. The result may be a reduction and/or attenuation of a signal, symptom or cause of a disease, or any other desired change in a biological system. An appropriate therapeutic amount for any individual can be determined by one skilled in the art using routine experimentation.

As used herein, the term "veterinarily acceptable" means that it does not interfere with the biological activity or properties of the compound and is relatively non-toxic, that is, the material does not have an undesirable biological effect or any component of the composition it contains. Refers to a substance, such as a carrier or diluent, that can be administered to an individual without interacting in a detrimental way with the component.

As used herein, the term "veterinarily acceptable salt" refers to a salt of a compound to be administered prepared from non-toxic acids including inorganic acids, organic acids, solvates, hydrates or clathrates thereof that are acceptable for veterinary medicine. indicate Examples of such inorganic acids are hydrochloric acid, hydrobromic acid, iodic acid, nitric acid, sulfuric acid, phosphoric acid, acetic acid, hexafluorophosphoric acid, citric acid, gluconic acid, benzoic acid, propionic acid, butyric acid, sulfosalicylic acid, maleic acid, lauric acid, malic acid, fumaric acid. , succinic acid, tartaric acid, amsonic acid, palmic acid, p-toluenesulfonic acid and mesylic acid. Suitable organic acids may be selected, for example, from the aliphatic, aromatic, carboxylic and sulfonic classes of organic acids, such as formic acid, acetic acid, propionic acid, succinic acid, camphorsulfonic acid, citric acid, fumaric acid, gluconic acid, isethionic acid, Lactic acid, malic acid, mucous acid, tartaric acid, para-toluenesulfonic acid, glycolic acid, glucuronic acid, maleic acid, furoic acid, glutamic acid, benzoic acid, atranilic acid, salicylic acid, phenylacetic acid, mandelic acid, emphonic acid (pam acid), Examples include methanesulfonic acid, ethanesulfonic acid, pantothenic acid, benzenesulfonic acid (besylate), stearic acid, sulfanilic acid, alginic acid, and galacturonic acid. Moreover, veterinarily acceptable salts include, but are not limited to, alkaline earth metal salts (eg calcium or magnesium), alkali metal salts (eg sodium-dependent or potassium) and ammonium salts. includes

As used herein, the term "veterinarily acceptable carrier" means a liquid or solid filler related to transport or transport so that chlortetracycline can perform its intended function in or to a mammal within the present invention. , a veterinarily acceptable substance, composition or carrier such as a stabilizer, dispersant, suspending agent, diluent, additive, thickening agent, solvent or encapsulating material. Typically, these components are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in terms of being compatible with other ingredients of formulas including compounds useful within the present invention and not injurious to mammals. Some examples of materials that can serve as veterinary acceptable carriers include: sugars such as lactose, glucose and sucrose used in veterinary formulations; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethylcellulose and cellulose acetate; tragacanth powder; malt; gelatin; talc; additives such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; Surfactants; alginic acid; pyrogen-free water; isotonic solution; Ringer's solution; ethyl alcohol; phosphate buffer solution; and other non-toxic compatible substances. In addition, as used in the present invention, "veterinary acceptable carrier" is compatible with the activity of compounds useful within the present invention, and is physiologically acceptable to mammals in part or in whole coating, antiviral and antibacterial agents and delayed absorption. including my back Supplementary active compounds may also be incorporated into the compositions. Further "veterinarily acceptable carriers" may further include veterinarily acceptable salts of compounds useful within the present invention. Other additional ingredients that may be included in veterinary compositions used in the practice of the present invention are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co. , 1985, Easton, PA).

As used herein, the term "composition" or "veterinary composition" refers within the present invention to a mixture of chlortetracycline and a veterinarily acceptable carrier. The veterinary composition facilitates administration of chlortetracycline to a mammal or subject. A variety of techniques exist in the art for administering compounds including, but not limited to, intravenous, oral, aerosol, parenteral, ocular, pulmonary and topical administration.

The veterinary composition according to the present invention may further include suitable excipients and diluents commonly used in the preparation of veterinary compositions.

The veterinary dosage forms of the present invention may be used in the form of their veterinarily acceptable salts, and may also be used alone or in combination with other veterinarily active compounds as well as in suitable combinations.

Excipients and diluents that may be included in the veterinary composition according to the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium Silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin, sorbitan mono stearate, polysorbate 60, methylparaben, propylparaben and mineral oil.

The veterinary composition according to the present invention may further include fillers, anti-agglomerating agents, lubricants, wetting agents, spices, emulsifiers, preservatives, etc. Or it may be formulated using a method well known in the art to provide delayed release, and the dosage form may be a powder, granule, tablet, capsule, suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, It may be in the form of a suppository, a sterile injection solution, or a sterile external preparation.

The veterinary composition according to the present invention may vary depending on the age, sex, and weight of the animal, but may be administered in an amount of 0.1 to 100 mg/kg once to several times a day, and the administration period may also be sustained for several weeks to several months. there is. In addition, it may increase or decrease according to the route of administration, severity of disease, gender, weight, age, etc. Accordingly, the dosage is not intended to limit the scope of the present invention in any way.

In addition, the present invention provides an animal feed additive useful for preventing and improving FIP containing chlortetracycline or a veterinarily acceptable salt thereof as an active ingredient and a feed containing the same.

The above composition for animal feed additives may be in the form of 20 to 90% high concentration or powder or granular form.

The animal feed additive of the present invention contains organic acids such as citric acid, fumaric acid, adipic acid, lactic acid and malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polyphosphate), polyphenol, catechin, Any one or more than one of natural antioxidants such as alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid may be further included.

The animal feed additive of the present invention and the feed containing the same are supplementary ingredients, such as amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal enzymes, and live microbial agents, such as grains, for example, crushed or crushed. cooked wheat, oats, barley, corn and rice; vegetable protein feeds such as those based on rape, soybean and sunflower; animal protein feed such as blood meal, meat meal, bone meal and fish meal; A dry ingredient consisting of sugar and dairy products, such as various powdered milk and whey powder, and dry additives are all mixed, and then a liquid component and a component that becomes liquid after heating, that is, a lipid, such as an animal product optionally liquefied by heating In addition to main ingredients such as fat and vegetable fat, it can be used with substances such as nutritional supplements, digestion and absorption enhancers, growth promoters, and disease preventive agents.

The animal feed additive may be administered alone to the animal in combination with other feed additives in an edible carrier.

In addition, the animal feed additives can be easily administered as a top dressing or directly mixed with animal feed, separately from feed, in a separate oral formulation, by injection or transdermally, or in combination with other ingredients. Typically, a single daily dose or divided daily doses may be used, as is well known in the art.

When the animal feed additive is administered separately from the animal feed, as is well known in the art, the dosage form of the extract can be prepared as an immediate release or sustained release formulation by combining them with a non-toxic pharmaceutically acceptable edible carrier. Such edible carriers can be solid or liquid, for example corn starch, lactose, sucrose, corn flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the dosage form of the extract may be tablets, capsules, powders, troches or sugar-containing tablets, or top dressing in a microdispersible form. When a liquid carrier is used, it may be a soft gelatin capsule, or a dosage form of a syrup or liquid suspension, emulsion or solution. In addition, the dosage form may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like. In addition, they may contain other substances useful for the improvement and prevention of diseases caused by viruses.

In addition, the animal feed included in the animal feed additive according to the present invention may be any protein-containing organic flour commonly used to meet the dietary needs of animals. These protein-containing grains usually consist mainly of corn, soybean flour or corn/soybean flour mixes.

The animal feed additive may be used by adding it to the animal feed by dipping, spraying or mixing.

In the animal feed formulation method including the extract according to the present invention, the animal feed additive is incorporated into the animal feed in an amount of about 1 g to 100 g per 1 kg of feed on a dry weight basis.

Further, after the feed mixture is thoroughly mixed, a hard, granular or granular material is obtained depending on the grinding degree of the components. It is supplied as a mash or formed into desired discrete shapes for further processing and packaging. At this time, in order to prevent separation during storage, it is preferable to add water to the animal feed and then to subject it to a conventional pelletization, expansion, or extrusion process. Excess water may be dried off.

The present invention also provides a mammalian health food composition for preventing or improving feline infectious peritonitis comprising chlortetracycline or a veterinarily acceptable salt thereof as an active ingredient.

The health food composition of the present invention may be tablets, capsules, pills, powders or liquids for mammals, and may be consumed by itself or added to various foods or beverages at an amount of 0.01 to 100% by weight.

The present invention also provides a method for treating feline infectious peritonitis, comprising administering chlortetracycline or a veterinarily acceptable salt thereof to a non-human mammal.

The present invention also provides an antiviral composition against feline infectious peritonitis virus comprising chlortetracycline or a veterinarily usable salt thereof.

The antiviral composition according to the present invention may be a quasi-drug composition.

The quasi-drug composition of the present invention is not limited thereto, but is preferably disinfectant cleaner, shower foam, gargreen, wet tissue, detergent soap, hand wash, humidifier filler, bowel pad, disinfectant spray, mask, ointment, patch, or filter filling It can be the first, which is used to remove and disinfect the feline infectious peritonitis virus, and can be used for the purpose of preventing mammals from being infected with the feline infectious peritonitis virus and for the purpose of phase reproduction.

The composition of the present invention comprising chlortetracycline or a veterinarily acceptable salt thereof inhibits the enzymatic activity of feline infectious peritonitis virus and has a very high activity to inhibit the virus from proliferating intracellularly and forming plaques, thereby preventing feline infectious disease. It can be very useful for preventing peritonitis or developing a therapeutic agent.

1 is a diagram showing that the enzyme activity of FIPV virus is significantly inhibited by chlortetracycline and tetracycline.
Figure 2 is a graph showing plaques (plaque) formed by viral growth after infection with feline infectious peritonitis virus in a cat cell line (A), and quantification thereof (B).
3 is a diagram showing the results of blood analysis of normal cats subcutaneously administered with 50 mg/kg of chlorotetracycline, twice a day, for a total of 5 days (3a: 2 years old, 3b: 8 months old)
Figure 4 shows X-rays taken before subcutaneous administration of chlorotetracycline at 20 mg/kg twice a day for a total of 5 days to cats with feline infectious peritonitis, 1 week after administration and 2 weeks after administration, and visually observed clinical findings. is a result
Figure 5 shows WBC, Globulin and A/G ratio obtained by obtaining blood before, 1 week after administration and 2 weeks after administration of chlorotetracycline at 20 mg/kg twice a day, subcutaneously for a total of 5 days in cats with feline infectious peritonitis. This is the result of analyzing the values.

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto.

Example 1: Chlortetracycline's ability to inhibit FIPV enzyme activity

After obtaining the sequence of the gene expressing the feline infectious peritonitis virus proteolytic enzyme from NCBI, the sequence was chemically synthesized and cloned into a pBT vector. Using this plasmid, the FIPV enzyme was isolated from Escherichia coli, and the FIPV enzyme reaction was performed using this plasmid.

The cleavage enzyme reaction was performed under conditions of 20 nM Tris-HCL (pH 7.5), 200 mM NaCl, 0.2 mM EDTA, 0.2 mM DTT, and 1% DMSO, and was reacted for 4 hours with a mixture of 2 μM protein and 50 μM fluorescent substrate.

The results for this are shown in Figure 1.

As shown in Figure 1, the IC50 of chlortetracycline's ability to inhibit FIPV enzyme activity was about 17 μM, but tetracycline was 80 μM, confirming that chlorotetracycline was more than four times effective in inhibiting enzyme activity.

Example 2: Inhibiting ability of chlortetracycline to form viral plaques

Negative control, positive control, and samples treated with chlortetracycline were infected with 100-fold diluted supernatant obtained after viral infection.

In this experiment, Feline Enteric Peritonitis Virus (FEPV), which can be used as a model for feline infectious peritonitis virus (FIPV), was used for safety reasons. After incubating these samples in a 37 degree incubator for 1 hour, 2xMEM overlay media and 0.6% agar were mixed in a 1:1 ratio and dispensed at 1.5 ml per well.

After 3 days, it was fixed with 4% paraformaldehyde for 1 hour. Thereafter, crystal violet was treated and stained, and plaque formation was observed and counted.

The results for this are shown in Figure 2.

As shown in FIG. 2 , it was confirmed that plaque formation due to virus propagation was reduced in a concentration-dependent manner in the cat cell line treated with chlortetracycline.

Example 3: Safety evaluation of chlortetracycline

To evaluate the safety of chlorotetracycline, a formulation containing chlortetracycline at a concentration of 50mg/ml was prepared, and 50mg/kg was administered twice a day (12 hours apart) for 5 days to normal subject cats (n=2, 8 months old). , 2 years of age) was administered subcutaneously (SC). After administration was completed, blood was obtained from each animal and blood analysis was performed.

The results for this are shown in Figures 3a and 3b.

As shown in FIGS. 3A and 3B , it was confirmed that no particular abnormality was observed as a result of blood analysis of each animal administered with chlorotetracycline.

Example 4: Efficacy evaluation of chlortetracycline

To evaluate the treatment effect of feline infectious peritonitis virus infection of chlorotetracycline, feline infectious peritonitis patients (8-9 months old, female, 3.3 kg) were given tetrachlorocycline at 20 mg/kg twice a day (12 hours apart) for a total of It was administered subcutaneously for 5 days, and clinical findings, blood analysis, and X-ray results were observed for 2 weeks after the end of administration.

As a result, radiological observation showed that pleural effusion and ascites abnormalities were not found for 2 weeks after administration (FIG. 4), and it was confirmed that the symptoms of feline infectious peritonitis were improved based on blood levels (FIG. 5).

The composition of the present invention comprising chlortetracycline or a veterinarily acceptable salt thereof inhibits the enzymatic activity of feline infectious peritonitis virus and has a very high activity to inhibit the virus from proliferating intracellularly and forming plaques, thereby preventing feline infectious disease. It can be very useful for preventing peritonitis or developing a therapeutic agent, so industrial applicability is very high.

Claims (13)

A veterinary composition for preventing or treating feline infectious peritonitis (FIP), comprising chlortetracycline or a veterinarily acceptable salt thereof as an active ingredient.
The veterinary composition according to claim 1, wherein the veterinary subject is a cat.
The veterinary composition according to claim 1, wherein the veterinary composition is for oral, intravenous, intraperitoneal, intramuscular or subcutaneous administration.
An animal feed additive for preventing or treating infectious peritonitis in cats, comprising chlortetracycline or a veterinarily acceptable salt thereof as an active ingredient.
An animal feed comprising the animal feed additive according to claim 4.
The feed according to claim 5, characterized in that the feed is for cats.
A mammalian health food composition for preventing or improving feline infectious peritonitis, comprising chlortetracycline or a veterinarily acceptable salt thereof as an active ingredient.
The health food composition according to claim 7, wherein the mammal is a cat.
A method for treating feline infectious peritonitis, comprising administering chlortetracycline or a veterinarily acceptable salt thereof to a mammal other than humans.
10. The method of claim 9, wherein the mammal is a cat.
An antiviral composition against feline infectious peritonitis virus comprising chlortetracycline or a veterinarily acceptable salt thereof.
The antiviral composition according to claim 11, wherein the antiviral composition is a quasi-drug composition.
[Claim 13] The antiviral composition according to claim 12, wherein the quasi-drug is selected from the group consisting of disinfectant cleaner, shower foam, wet tissue, detergent soap, humidifier filler, toilet pad, and disinfectant spray.

Images