KR20170132566A - A veterinary composition comprising sustained release antibiotics formulation and a manufacturing method for the same - Google Patents

Abstract

A sustained release veterinary composition for prevention or treatment of a disease caused by bacteria and a manufacturing method thereof of the present invention, can provide a method for manufacturing a veterinary composition comprising antibiotics capable of mass-produced in a relatively simple manner by chemically bonding an antibiotic active ingredient to a peroxidized fatty acid, and a veterinary composition which can be used as sustained-release injections. The veterinary composition may have drug persistence of 60 hours or greater after injection into an animal subject to be applied in a relatively simple manner to diseases caused by bacteria, and to reduce the labor required for treatment. In addition, it is possible to reduce the stress which an animal can receive in the process to improve a growth rate, feed efficiency and the like as well as an effect of treating bacterial diseases due to medicines as compared with a conventional treatment method.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a sustained-release veterinary composition of antibiotics and a method for producing the same.

The present invention relates to a veterinary composition comprising an antibiotic component which can be used as a sustained-release injection.

The sustained-release injection formulation refers to an injection formulation that is formulated so that the drug can be continuously (uniformly) released (sustained release) while maintaining the biological activity in the body at the time of injection.

Conventionally, conventional methods for producing such sustained-release formulations are known as a coacervation method, a melt injection method, a spray drying method, a solvent evaporation method, and the like. Among these methods, a solvent evaporation method classified into a dual emulsion evaporation method (W / O / W emulsion) and a single emulsion evaporation method (O / W emulsion) is most widely used. However, such multi- , There is a disadvantage in that when the active ingredient drug is dissolved in an organic solvent or water, there are disadvantages such as changes in the physical properties of the drug, loss of stability, and low sealing efficiency while forming microspheres.

Various attempts have been made to increase the rate of drug encapsulation of such sustained-release formulations or to produce microparticles which are easy to manufacture and have relatively low initial over-release, but are still unsatisfactory.

(1983) 62-74), microspheres were prepared by double emulsion evaporation method, and then the active ingredient, vaccine antigen, was loaded through micropores of microspheres And then the micropores are blocked by heating at a temperature higher than the glass transition temperature (Tg) of the biodegradable polymer. However, such a preparation method has a serious disadvantage that the drug can be denatured by heat. In addition, the addition of a protein trapping agent significantly increased drug encapsulation efficiency (~97%), but the total encapsulated amount of active material (1.4 ~ 1.8%) was significantly lower. This method has a disadvantage in that the physiologically active substance can be denatured by heat, and there is an improvement in that it is difficult to encapsulate the drug in a large dose. In addition, there is a problem that the manufacturing process is somewhat complicated.

Quinolone antibiotics are antibiotics that work extensively on Gram-positive and Gram-negative bacteria.

(9-fluoro-2,3-dihydro-3-methyl-10- (4-methyl-1-piperazinyl) -7-oxo- (4,2,1) benzo-adiazine-6-carboxylic acid) is the most recently developed third-generation fluoroquinolone antibiotic and has a broad spectrum. It has a spectrum of gram-positive and gram- It works extensively. In particular, it acts effectively against Gram-negative bacteria and functions to slow the production of strains by acting on two target protein DNAs, gyrase and topoisomerase IV, of bacteria.

Quinolone drugs are prepared in colloidal form and injected into the animal. In general, the suspending agent should have properties such as therapeutic efficacy, physical and chemical stability of the pharmaceutical ingredient, and durability of the pharmaceutical preparation. However, currently available quinolone pharmaceutical preparations have a disadvantage of requiring frequent administration because of their short duration, There is a problem that the sedimentation speed of the particles is so fast that it is difficult to reproduce.

Prior art relating to a suspension injecting agent is a medicine containing fluoroquinolones using an ammonium compound (WO 2008-025380) or an aqueous composition using nicotinamide as an adjuvant (Korean Patent Registration No. 10-0106603). There is also stabilization of oily suspensions comprising hydrophobic silica (WO 2009-065514).

It is an object of the present invention to provide a veterinary composition for an antibiotic which can be used as a sustained-release injection, and specifically to provide a veterinary composition which can be used as a sustained-release formulation for administration of an antibiotic component capable of binding to peroxide.

In order to accomplish the above object, the present invention provides a method of preparing a sustained veterinary composition for preventing or treating diseases caused by bacteria in an animal, which comprises adding an oxidized unsaturated fatty acid to a peroxide value of not less than 300 meq / Preparing a peroxyated oil to prepare a mixture for supporting an antibiotic by mixing the peroxyated oil, which is an oil containing the oil, with a stabilized oil containing a saturated oil; And an antibiotic agent containing the sustained-release antibiotic-bearing oil in which the antibiotic component is chemically bonded to the peroxide oil by mixing the antibiotic-containing mixture with the antibiotic component containing a carboxyl group, an amine group or both in the molecule, And a binding step of preparing the pharmaceutical preparation.

Wherein the peroxide oil preparation step comprises: an oxidation step of oxidizing an oil containing an unsaturated fatty acid to produce a peroxide oil having a peroxide value of 300 meq / kg or more; And a stabilization step of mixing a stabilized oil containing a saturated oil with the peroxide oil to prepare a mixture for supporting antibiotics.

The oxidation of the oxidation step may be carried out by an air injection method of injecting oxygen-containing air into the oil containing the unsaturated fatty acid.

The oil containing the unsaturated fatty acid may have a polyunsaturated fatty acid content of 50 wt% or more based on the total oil.

The oil containing the unsaturated fatty acid may be any one selected from the group consisting of a combination of a combination thereof, such as a toothpaste oil, a kiwi seed oil, a perilla seed oil, a flaxseed oil, and combinations thereof.

The oxidation step may be carried out at an oxidation temperature of 70 to 120 ° C.

The antibiotic component may include any one selected from the group consisting of marbloxacin, danofloxacin, enrofloxacin, their respective veterinarily acceptable salts, and combinations thereof as an active ingredient.

In the stabilization step, the stabilizing oil may be applied at 50 to 99% by volume based on the sum of the peroxide oil and the stabilizing oil.

The content of the antibiotic component contained in the sustained release antibiotic-supporting oil may be less than 300 mg / mL.

The composition for preparing a sustained-release preparation for the prevention or treatment of diseases caused by bacteria in an animal according to another embodiment of the present invention is a peroxide oil having a peroxide value of 300 meq / kg or more; A stabilizing oil containing a saturated oil; And an antibiotic active ingredient comprising a carboxyl group, an amine group or both in the molecule.

A sustained release formulation of antibiotics comprising a stabilized oil containing a sustained-release antibiotic-bearing oil and a saturated oil comprising an antibiotic component chemically bonded to a chain of fatty acids according to another embodiment of the present invention .

The sustained-release formulation of the antibiotic may contain up to 300 mg / mL of the antibiotic component and may have a viscosity of less than 280 cP (20 ° C).

The sustained-release preparation of the antibiotic may be one having a drug persistence of 60 hours or more after the injection.

A method of treating an animal other than a human according to another embodiment of the present invention includes a step of injecting a sustained veterinary composition for prevention or treatment of a disease caused by the above-described bacteria.

Hereinafter, the present invention will be described in more detail.

In this specification, the sustained-release preparation is based on an oxidized oil complex formed by combining an antibiotic component and a peroxide oil, and has a high amount of encapsulated drug in the oil phase and a high encapsulation efficiency. After the administration, Or an agent exhibiting a drug release behavior that continuously releases a drug for a predetermined period in such a manner that an antibiotic component is absorbed into capillary blood vessels due to the breakdown of the binding between the antibiotic component and the peroxide.

The term " treatment " as used herein means to alleviate or eliminate a disease (clinical symptom) caused by a bacterium by administration of the composition of the present invention. For example, Or amelioration or alleviation of symptoms or conditions such as fever, inflammation, coughing, diarrhea, reduction of symptoms, stabilization of symptoms, non-proliferation, suppression of symptoms, delay or delay of symptom progression, And may include a decline in symptom development.

In describing the present invention, the singular forms of the present invention are not limited to singular and plural, except for the case where it is obvious that the singular form is singular in the context of the present invention. do.

Or " and / or "are used interchangeably and the description of" A or B "or" A and / or B " Can be interpreted as including meaning.

The term "veterinarily acceptable salt thereof" or "its salts ", as used herein in connection with a particular compound, refers to a compound that possesses the biological effectiveness and properties of parent compounds, Quot; means salts which, when administered, are not deleterious to the biologically or otherwise. For example, the above "veterinarily acceptable salt thereof" or "a salt thereof" may be a base addition salt of a specific compound and may be prepared from inorganic and organic bases. Salts derived from inorganic bases may include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines; Substituted amines including naturally occurring substituted amines; And organic amines such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, The compounds of the present invention can be used in the form of hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamine, theobromine, purine, piperazine, piperidine, Lt; RTI ID = 0.0 > of Cyclic < / RTI > In addition, carboxylic acid amides, including other carboxylic acid derivatives such as carboxamides, lower alkyl carboxamides, di (lower alkyl) carboxamides, and the like, may also be useful in the practice of the invention have.

A method for preparing a sustained veterinary composition for the prevention or treatment of a disease caused by bacteria in an animal other than a human according to an embodiment of the present invention is an oil containing an unsaturated fatty acid oxidized to have a peroxide value of 300 meq / Preparing a peroxide oil to prepare a mixture for supporting antibiotics by mixing the peroxide oil with a stabilizing oil containing a saturated oil; And an antibiotic agent containing the sustained-release antibiotic-bearing oil in which the antibiotic component is chemically bonded to the peroxide oil by mixing the antibiotic-containing mixture with the antibiotic component containing a carboxyl group, an amine group or both in the molecule, And a binding step of preparing the pharmaceutical preparation.

Wherein the peroxide oil preparation step comprises: an oxidation step of oxidizing an oil containing an unsaturated fatty acid to produce a peroxide oil having a peroxide value of 300 meq / kg or more; And a stabilization step of mixing a stabilizing oil containing a saturated oil with the peroxide oil to prepare a mixture for supporting antibiotics. At this time, the oxidation of the oil containing the unsaturated fatty acid and the mixing of the stabilizing oil may be carried out in the order of oxidation after mixing in a different order.

The oxidation step is a step of activating an unsaturated fatty acid so as to be chemically bonded to a functional group such as a carboxyl group, an amine group or a ketone group contained in the molecule of the antibiotic component in a subsequent binding step to the oil containing the unsaturated fatty acid, carbon - to a double bond of an unsaturated fatty acid comprising a carbon-carbon double bond reacts with oxygen (O ₂₎ hydrogen ^{peroxide-group-is} the process of forming (e.g., -OOH, -OO, -OH, etc.). Such peroxygenated oils can form ester bonds and / or amide bonds by binding to the functional groups contained in the molecule of the above antibiotic component, such as a peroxide group or a carboxyl group of a fatty acid.

At this time, the activation of the unsaturated fatty acid may proceed in the course of oxidation, and a method of supplying oxygen to the unsaturated fatty acid may be applied. Specifically, a method of supplying air containing oxygen to the oil containing the unsaturated fatty acid may be applied (Air sparging, O ₂ sparging).

The oil to be oxidized may be an oil containing an unsaturated fatty acid. Specifically, the oil may include an unsaturated fatty acid having at least one double bond in the molecule such as linoleic acid, linolenic acid, oleic acid, methacrylic acid, arachidonic acid, And an unsaturated fatty acid having an even number of carbon atoms having 12 to 20 carbon atoms can be applied. Specific examples of the unsaturated fatty acids include oleic acid having two double bonds in the molecule, and the like. The unsaturated fatty acids include those having two or more double bonds in the molecule, such as linoleic acid having two double bonds in the molecule and linolenic acid having three double bonds in the molecule The application of an oil containing a polyunsaturated fatty acid is advantageous in that it can delay the mass release of the initial drug.

Specifically, the oil containing the unsaturated fatty acid can be applied so that the content of the polyunsaturated fatty acid is 50% by weight or more based on the total oil. More specifically, in the oil containing the unsaturated fatty acid, the sum of the content of linoleic acid and the content of linolenic acid may be 50% by weight or more based on the total oil, and the sum of the content of linoleic acid and the content of linolenic acid may be Based on the total oil, and the content of the phenolic acid is 50% by weight or more based on the whole oil.

In particular, when an oil containing at least 50% by weight of an unsaturated fatty acid having three or more double bonds in the molecule is applied as an oil containing an unsaturated fatty acid in the oxidation step, the sustained release properties of the antibiotic A well-controlled western-direction antibiotic-bearing oil can be produced.

The oxidation process may be performed at a relatively high temperature by heating the oil containing the unsaturated fatty acid. Specifically, the oxidation process may be carried out at an oxidation temperature of 70 to 120 ° C, Lt; RTI ID = 0.0 > 85-105 C. < / RTI >

Specifically, when flaxseed oil is used as the oil containing the unsaturated fatty acid, the oxidation process is improved by injecting air containing oxygen or oxygen while maintaining the oxidation temperature of 95 to 105 ° C. .

The time for maintaining the oxidation temperature in the oxidation step may be, for example, 0.5 to 15 hours, may be for 2 to 8 hours, and may be controlled by measuring the peroxide change in the oil containing the unsaturated fatty acid .

Peroxidized oils that have undergone this oxidation step can have a peroxide value of greater than 300 meq / kg, greater than 400 meq / kg, and greater than 450 meq / kg. The peroxide value of the peroxide oil may be from 300 meq / kg to 1500 meq / kg, from 300 meq / kg to 1200 meq / kg, and from 400 meq / kg to 1100 meq / kg.

The peroxide value of the oxidized peroxide can be used as an index showing how much the chemical bond between the active ingredient of the antibiotic and the oil component proceeds at a later stage. However, when the peroxide value is high, the viscosity of the oil may be too high Which may lead to restrictions on the use of the veterinary composition of the present invention. That is, the administration of the veterinary composition of the present invention does not have too high a viscosity to be carried out using a conventional syringe, and at the same time, it contains a high content of antibiotic active ingredient per unit volume (or unit weight) It is necessary to have such a peroxide value as mentioned above.

The peroxide value is a criterion for confirming whether the double bond in the unsaturated fatty acid is sufficient for binding the antibiotic component, and the oil containing the unsaturated fatty acid is sufficiently oxidized, and can be measured by a normal peroxide measurement method.

Specifically, 1 g of the sample was dissolved in 25 mL of a mixed solution of acetic acid / chloroform (3: 2), and 1 mL of the saturated KI solution prepared was added thereto. After thorough mixing, the mixture was allowed to stand in a dark place for 10 minutes, And the mixture is shaken. The solution is titrated with a 0.01 N Na ₂ SO ₃ .5H ₂ O solution and then blanked separately. The corrected value (indicator: starch solution) is then calculated.

Peroxide value (unit: meq / kg) = {(a-b) x f) / S x 10

a: The amount of consumption of _{0.01N Na 2 SO 3 · 5H 2} O solution of the test _{(mL) b: 0.01N Na 2} SO of the blank test is carried out ₃ · 5H ₂ O consumption of solution _{(mL) f: 0.01N Na 2} SO 3 · 5H ₂ O Potency of solution S: Weight of sample (g)

When the natural oil derived oil is applied to the oil containing the unsaturated fatty acid, any one selected from the group consisting of, for example, a toothpaste oil, a kiwi seed oil, a perilla seed oil, a flaxseed oil and a combination thereof may be applied. These oils are relatively high in the content of the lanolin acid, so that the increase of the peroxide of the oil is comparatively easy due to the oxidation method described above, and the sustained-release characteristics of the sustained-release antibiotic-supported oil produced thereafter are relatively good.

Particularly, in the case of olive oil having a large amount of monounsaturated fatty acids or the content of polyunsaturated fatty acids, the content of linolenic acid is relatively higher than that of cottonseed oil, which is mostly linoleic acid, There is also an advantage that the amount of the antibiotic component that is initially released during the production of the sustained-release antibiotic-supported oil is relatively small. However, if the peroxide value is excessively high, the viscosity may become too high. However, it is possible to control the viscosity and the like of the content of the stabilized oil to be mixed in the stabilization step, meq / kg to 1,500 meq / kg.

Wherein the stabilizing step comprises mixing the stabilized oil containing the saturated oil with the peroxide oil to prepare an antibiotic-supporting mixture in which the peroxide oil is stabilized, minimizing unintended reaction by the peroxide group contained in the peroxide oil , Which helps to induce chemical binding with antibiotic components added later. The stabilizing oil may block or delay the progress of the reaction of the peroxide with the fatty acid and other components before the chemical bonding reaction between the antibiotic component added in the binding step and the peroxide group of the peroxide oil proceeds, The viscosity of the sustained-release antibiotic-bearing oil can also be controlled.

As the stabilizing oil, a saturated fatty acid, a neutral fat, etc. may be applied. Specifically, a fatty acid which is not oxidized by oxygen and is connected to a single bond (C16: 0, C1: 0) For example, medium chain triglycerides (Miglyol) extracted from milk fat or coconut oil may be utilized.

In the stabilization step, the content of the stabilizing oil may be 50 to 99% by volume, 60 to 99% by volume, 70 to 97% by volume, or 80 to 99% by volume when the sum of the peroxide- 97% by volume. In this case, the unstable chemical reaction which may be caused by the peroxide group of the peroxide can be sufficiently blocked or delayed by the above-mentioned amount of the stabilizing oil, and the sustained-release antibiotic- Can be controlled.

The combining step is a step of preparing a sustained-release preparation of an antibiotic containing a sustained-release antibiotic-supporting oil prepared by mixing the antibiotic-supporting mixture with an antibiotic component and chemically bonding the antibiotic component to the peroxide oil.

The antibiotic component (antibiotic active ingredient) contains a carboxyl group, an amine group or both functional groups in the molecule, and these functional groups are present in an active state in a chain of peroxide-containing oil contained in the mixture for supporting antibiotics Which can be chemically combined with the peroxide group.

As the antibiotic component, 6-fluoro-2,3-dihydro-3-methyl-10- (4-methyl-1-piperazinyl) -7-oxo-7H-pyridol (3,2,1-ij (4,2,1) benzoxadiazin-6 carboxylic acid, danofloxacin, enrofloxacin, their respective veterinarily acceptable salts, and combinations thereof. These components include carboxyl groups, amine groups, or both functional groups in the molecule. In the binding step, a bond such as a covalent bond is generated by the chemical reaction between the peroxide group of the peroxide and the functional group in the binding step. A sustained-release antibiotic-bearing oil (a sustained-release antibiotic-bearing fatty acid-containing oil) containing the antibiotic component suspended in a chemical bond may be formed.

At this time, it is considered that the binding includes a case where the antibiotic component and the fatty acid contained in the peroxide oil are covalently bonded to each other, but the physical property of the antibiotic component in which the antibiotic component is located in the space formed by the bonds between the fatty acids It is not meant to exclude other forms of chemical bonding, such as ionic or hydrogen bonding, and is not meant to exclude the presence of the antibiotic A component that is recognized to be bioequivalent to the active ingredient is slowly released, so that a binding that can detect a certain level or more in the plasma of the animal to which the sustained-release veterinary composition is applied suffices.

The content of the antibiotic component contained in the sustained-release antibiotic-bearing oil may be less than 300 mg / mL, may be 280 mg / mL or less, and may be 80 to 260 mg / mL. When the content of the antibiotic component contained in the sustained-release antibiotic-supporting oil is 300 mg / mL or more, it may not be easy to release the antibiotic component in the sustained-release antibiotic-supported oil itself in vivo .

The content of the peroxide oil contained in the sustained release preparation may be 300 mg or less based on 1 mL of the sustained release preparation, and may be 150 mg or less. If the content of the peroxide oil is excessive, the viscosity of the sustained-release antibiotic-coated oil itself may increase, which may make it difficult to use as an injection,

The sustained-release veterinary composition may have a viscosity of less than 280 cP (20 캜, centipoise), may be less than 230 cP (20 캜), less than 200 cP (20 캜), and may range from 100 to 200 cP. If the viscosity of the sustained-release veterinary composition is greater than 280 cP, release of the active ingredient from the sustained-release veterinary composition after intramuscular injection may be impaired.

The use of the sustained-release veterinary composition may be applied to the extent that biological equivalence with the antibiotic carried on the sustained-release antibiotic-bearing oil is recognized and may be used for the prevention or treatment of bacterial diseases. The bacterial disease may be bacterial pneumonia or bacterial enteritis. Specifically, the bacterial disease may be pleural pneumonia, Pasteurella pneumonia, Glucorhosis, or Escherichia coli. In addition, the bacterium that causes the bacterial disease is Actinobacillus ( Actinobacillus < RTI ID = 0.0 > pleuropneumoniae , Pasteurella but are not limited to, E. coli , C. multocida , Haemophilus parasuis , and E. coli .

The sustained-release veterinary composition for the prevention or treatment of a disease caused by a bacterium produced by the above-mentioned method is characterized in that it comprises a sustained-release antibiotic-bearing oil comprising an antibiotic component chemically bonded to a chain of a fatty acid, The effective ingredient of the antibiotic can be continuously released for more than 3 days or more than 5 days, including the sustained-release preparation containing the antibiotic including the oil, and the continuous treatment or prevention effect can be obtained while reducing the number of times of administration .

The description of the kinds, contents, characteristics, uses, etc. of the antibiotic component contained in the sustained-release preparation of the antibiotic is the same as that of the above-described preparation method, and thus description thereof will be omitted.

The sustained-release preparation of the antibiotic may be used as a injectable preparation, and may have drug persistence (a property of continuously releasing the drug over an effective concentration) for 60 hours or more after injection by intramuscular injection or the like, and 80 Drug persistence over time, and may have drug persistence above 120.

The composition for the preparation of a sustained-release preparation for the prevention or treatment of diseases caused by bacteria in an animal according to another embodiment of the present invention is a peroxide oil having a peroxide value of 300 meq / kg or more; A stabilizing oil containing a saturated oil; And an antibiotic active ingredient comprising a carboxyl group, an amine group or both in the molecule. Detailed description of the oil containing the unsaturated fatty acid applied to the production of the peroxide oil, or the peroxide oil, the stabilizing oil, the antibiotic active ingredient, etc. prepared therefrom will be omitted.

The sustained-release veterinary composition for the prevention or treatment of diseases caused by bacteria in an animal according to another embodiment of the present invention comprises a sustained-release antibiotic-bearing oil comprising an antibiotic component chemically bonded to a chain of a fatty acid, A sustained release formulation of an antibiotic comprising a stabilizing oil containing a saturated oil.

The sustained-release preparation of the antibiotic may include a fluoride-quinolone antibiotic component-bearing oil, and a saturated oil. The fluoride-quinolone antibiotic component is chemically bonded to the peroxide-containing oil by covalent bonding to form the fluorinated quinolone antibiotic Component-bearing oil can be constituted.

Explanation of the peroxide value (unsaturation degree) of the peroxide oil, application amount of peroxide oil and saturated oil, content and content of the antibiotic component, viscosity of the sustained-release preparation of the antibiotic, duration in vivo , The description thereof will be omitted.

The veterinary composition may additionally comprise a pharmaceutical (or veterinarily) acceptable carrier in addition to the sustained release formulations described above.

The veterinary composition comprises a pharmaceutically effective amount of an antibiotic-extended formulation. As used herein, the term "pharmaceutically effective amount" refers to the amount of active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, an animal, as contemplated by a researcher, veterinarian, physician or other clinician, Lt; RTI ID = 0.0 > a < / RTI > disease or disorder. Effective dose levels include the type of disease and severity of the subject (animal), age, sex, type of animal, activity of the drug, sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, Factors and factors well known in the medical arts.

The carrier is usually used at the time of formulation, and may be selected from lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone , Cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.

The veterinary composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutical (or veterinarily) acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

In the present invention, the sustained-release preparation and / or the veterinary composition can be administered orally or parenterally, and parenteral administration is preferred. In the case of parenteral administration, it can be administered by intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection, transdermal administration, etc. and muscle injection is preferable.

The appropriate dosage of the sustained release formulation and / or the veterinary composition according to the present invention may vary depending on factors such as the formulation method, the mode of administration, the age, weight, sex, pathological condition, food, administration time, route of administration, And responsiveness. ≪ / RTI >

The sustained-release preparation and / or the veterinary composition according to the present invention can be prepared by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those skilled in the art. Or by formulating it into a unit dose form or by inserting it into a multi-dose container. Here, the formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The sustained release formulation and / or the veterinary composition is an antibiotic composition and may also be used as an injection. The sustained-release preparation and the veterinary composition containing the sustained-release preparation may be used for administration of domestic animals such as pigs, cows, horses, sheep, or poultry.

The method for treating an animal other than a human according to another embodiment of the present invention includes a step of injecting a sustained veterinary composition for prevention or treatment of a disease caused by the above-described bacteria. The description of the sustained-release veterinary composition is as described above. When the method is used to prevent or treat a bacterial disease, the method of injecting the composition once or twice at intervals of 5 days to 7 days Injection, etc.) can provide the antibiotic component to a therapeutic or prophylactic object, and it is possible to perform continuous and continuous antibiotic treatment in a simple manner in comparison with the existing method of daily injecting, and also to reduce the stress on the injection of the animal The amount of feed intake, the amount of gain, and the like can be minimized.

The present invention relates to a sustained-release veterinary composition for the prevention or treatment of diseases caused by bacteria and a method for producing the same, which comprises a method of producing a veterinary composition containing antibiotics of a sustained- Composition can be provided. Such a veterinary composition can have a drug persistence of 60 hours or longer after injection into a target animal, so that it can be applied relatively easily to a disease caused by a bacterium, and the labor required for treatment can be reduced. In addition, it is possible to reduce the stress that an animal can receive in the course of treatment, so that it is possible to improve the growth rate, feed efficiency, and the like, as well as the therapeutic effect of a bacterial disease caused by a drug.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the results of measurement of oxidation degree by oil in Experimental Example 1, which is an embodiment of the present invention,
FIG. 2 is a graph showing drug release behavior according to the change in peroxide value of flaxseed oil in Experimental Example 3 among the examples in which peroxide level per oil was measured according to the present invention.
FIG. 3 is a graph showing the results of in vitro drug release behavior measurement of an oil blend formulation in Experimental Example 1, which tested the in vitro drug release behavior of the present invention. FIG.
FIG. 4 is a graph showing the average concentration of flocosarin in the plasma among the results of analysis of the blood concentration of Marbofloxacin by time in the examples in which the in vivo drug activity and release behavior using pigs were tested in the present invention.
5 to 7 are graphs showing PK / PD analysis results of Tests 1 to 3 in Examples of the present invention, respectively.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

< Example : By oil Peroxidation  Measurement>

Experimental Example  1: Oxidation measurement by oil

Take 1 g of the sample and dissolve in 25 mL of a mixture of acetic acid and chloroform (3: 2). Add 1 mL of the prepared saturated KI solution, mix well, place in a dark place for 10 minutes, add 30 mL of distilled water and shake. The mixture was titrated with 0.01N Na ₂ SO ₃ .5H ₂ O solution, and the peroxide value was calculated using the following equation. Separately, a blank test was performed and corrected. Starch solution was used as an indicator.

Peroxide value (meq / kg) = {(a-b) x f) / S x 10

_{a: 0.01N Na 2 SO 3 ·} 5H 2 O solution of consumption (mL), b: 0.01N Na 2 SO 3 · 5H 2 O consumption of the solution of the blank test (mL), f: 0.01N Na 2 SO 3 · 5H ₂ O solution, S: Weight of sample (g)

As a sample, 100 ml of flaxseed oil, olive oil and cottonseed oil were directly injected into the oil at a rate of 1 L of high purity oxygen (95%) at 100 ° C and reacted for 8 hours or more to prepare a peroxide per hour oil sample The peroxide value was measured by the method of Fig. The contents of fatty acids contained in each oil are summarized in Table 1 below (% by weight).

vegetable oil Saturated fatty acid Monounsaturated fatty acid Polyunsaturated fatty acid Oleic acid Linolenic acid Linoleic acid Flaxseed oil 6-9 10-22 56-71 12-18 10-22 Cottonseed oil 25.9 17.8 One 54 19 Olive oil 14 72.0 1.5 9-20 -

1, in the case of olive oil, the degree of peroxidation did not increase even after prolonged oxidation, but in case of flaxseed oil, oxidation rapidly increased with time. When oxygen was injected under the same conditions, Respectively.

In olive oil, the content of monounsaturated fatty acids with one double bond in the molecular chain is 72%, which makes it difficult to increase the degree of peroxidation. In the case of cottonseed oil in which the content of linoleic acid (ω-6) with two double bonds in the molecule is 54% In the case of flaxseed oil, which contains 56-71% of linolenic acid (ω-3) with three double bonds in the molecule, this structural difference requires more time than other oils to show an intended constant degree of peroxidation. And the oxidation degree is increased. The degree of peroxidation of the peroxidized oils was varied according to the degree of unsaturation of the oil. In the next step, it affects the reaction with antibiotics such as magok flocasin, which is considered to be an important factor in determining the release characteristics.

Experimental Example  2: Confirmation of stability of oil formulation

The peroxide value of flaxseed oil (peroxide value: 978.24 mEq / kg) and cottonseed oil (peroxide value: 835.66 mEq / kg) were measured for about one month. The peroxide value of peroxide oil was maintained for more than one month within the error range, As a result, it can be seen that even when left at room temperature, the sustained release rate of the oil formulation is not changed.

Experimental Example  3: Estimation of Release Behavior with Change in Peroxide Value of Flaxseed Oil

The results of measurement of the release behavior according to the change in the peroxide value of linseed oil are shown in FIG. Flaxseed oil was prepared by mixing 5% (v / v) of microglycol. The higher the peroxide value, the lower the initial release, but the drug release over time was more efficiently controlled than the similarly tested cottonseed oil (data not shown for cottonseed).

Samples with a peroxide value of greater than or equal to about 400 mEq / kg had an initial release of less than about 10% and were found to release about 60% of the total drug within 3 days (72 hours). This shows that it has suitable characteristics as a slow release agent and it is a result of showing that the release rate can be controlled by the peroxide value of flaxseed oil. In particular, a sample with a peroxide value of greater than 1000 mEq / kg was found to release about 20% of the total drug within 3 days with a controlled maximum release rate.

< Example : In vitro drug release behavior test>

Example  One: Mabo Flocker Combined  Preparation of oxidized oil complex

Peroxide oil was prepared by adding high purity oxygen (95 v / v%) at 100 ℃. At this time, 1 liter of oxygen was added to 100 ml of flax seed oil, and the injection time was 8 hours or less. 9.5 ml of Miglyol medium chain triglyceride (5:95, v / v%) was added to 0.5 ml of the thus prepared peroxide oil (1277.9 (± 5.4) mEq / kg) and uniformly stirred to obtain a first mixed solution.

To the first mixed solution, 1 g of Mabofroxacin (Zhejiang Guobang Pharmaceutical Co., Ltd. .. micronized) was added to prepare a second mixed solution having a concentration of 100 mg / ml. The mixture was stirred for 6 hours by a magnetic stirrer to induce an ester reaction, A veterinary composition in the form of a sustained release formulation for injection of antibiotics was obtained.

Example  2: Mabo Flocker Combined  Preparation of oxidized oil complex

A veterinary composition, which is a sustained-release preparation for injection of maltodextroxin antibiotics, was obtained in the same manner as in Example 1, except that cottonseed oil was used instead of flaxseed oil in Example 1 above.

Comparative Example  One: Unoxidized  Manufacture using oil complex

A veterinary composition in the form of a sustained release formulation for injection was prepared in the same manner as in Example except for the use of unoxidized flax seed oil.

Comparative Example  2: Saturated oil  Manufacture using

1 g of Mabofroxacin (Zhejiang Guobang Pharmaceutical Co., Ltd .. micronized) was added to 10 ml of medium chain triglyceride (Miglyol) to prepare a mixed solution. The mixed solution was stirred for 6 hours with a magnetic stirrer to prepare a sustained- A medical composition was obtained.

The components and contents of the above Examples 1 and 2, Comparative Examples 1 and 2 are summarized in Table 2 below.

division Unsaturated oil (whether oxidized) Saturated oil Magoblockocaine Example 1 Flaxseed oil (oxidation) Miglyol 100 g / ml Example 2 Cottonseed oil (oxidation) Miglyol 100 g / ml Comparative Example 1 - Miglyol 100 g / ml Comparative Example 2 Flaxseed oil (non-oxidized) Miglyol ± 100 g / ml

Experimental Example  1: Measurement of in vivo drug release behavior of veterinary composition

The veterinary composition of Example 1, Example 2, Comparative Example 1, and Comparative Example 2 prepared above was placed in 30 ml of PBS (phosphate buffer saline, pH 7.4) in a total volume of 4 mg / ml, Lt; RTI ID = 0.0 &gt; C &lt; / RTI &gt; The collected samples were diluted 100-fold and placed in a disposable cuvette, and the absorbance at a wavelength of 335 nm was measured using a UVvis meter.

The absorbance value was converted into the concentration of the released drug, and the cumulative percentage of the released drug was calculated for each time period of each sample. The oil formulation prepared in Comparative Examples 1 and 2 was used as a control for this experiment. The measurement results are shown in Fig.

Referring to the results of FIG. 3, the drug was slowly released in Examples 1 and 2, and the results are shown in comparison with Comparative Example 1 and Comparative Example 2. In addition, in the case of Comparative Example 1, Comparative Example 2 and Example 2, the initial release of the drug was found to be higher than that in Example 1, and in the case of Example 1, the initial release amount was also effectively controlled. It was confirmed that the emission was effectively controlled. These results demonstrate that an oil-based sustained release formulation can be produced that can control the rate of drug release according to the degree of peroxidation of flaxseed oil and continuously release the drug over a period of three days or more. On the other hand, in the case of an oil in which the content of linolenic acid in the unsaturated oil is less than 50% (ω-3) (olive oil, cottonseed oil, etc.), the drug release over time is controlled to a certain extent, .

< Example : in vivo  Drug activity and release behavior test>

Manufacturing example  1: Sustainable Magoblockocaine  Preparation of veterinary compositions comprising the agent

Each of the peroxygenated unsaturated oils described in Table 3 below was prepared (Sparging) by oxidizing flaxseed oil at 100 ° C with air for about 6 hours to have a peroxide value of 460 meq / kg or more.

A part of saturated triglyceride, saturated triglyceride, was added to the peroxidized unsaturated oil and stirred. Then, marbophloxacin was slowly added while stirring, and then the remaining amount of trichlseride was added to prepare Tests 1 to 3. The contents of oils and antibiotics used in the manufacture of Tests 1 to 3, respectively, are shown in Table 3 below.

division Unsaturated oil application amount
(Degree of peroxidation, meq / kg) Saturated oil application amount
(mg / mL) Marbloxacin content
(mg / mL) Viscosity
(20 &lt; 0 &gt; C, cP) Test Approximately 1 150 mg / mL
(Over 1,000) Miglyol about 850 160 mg / ml 167.4 cP Test 2 150 mg / mL
(500) Miglyol about 850 250 mg / ml 182.4 cP Test Approx. 3 150 mg / mL
(500) Miglyol about 850 300 mg / ml 283.7 cP

(1) The above Tests 1 to 3 were each subjected to a content test after preparation and storage, and Test 1 was 152.277, 152.232, 152.843 and 152.451 mg / ml, respectively, and Test 2 was 246.561, 245.863, 254.074, 248.833 mg / ml, and test 3 were analyzed to contain active ingredients of 301.338, 299.167, 299.024, and 299.843 mg / ml, respectively.

2. Saturated oil is prepared by mixing approximately 80% by volume of the saturated oil application amount with peroxylated unsaturated oil, and administering the remaining amount after the reaction is carried out by administering malablockocin, wherein the total volume of the composition Was adjusted to be 1,000 mL.)

Example : Using pigs in vivo  Drug activity and release behavior test

<Experimental Method>

(Preparation of experimental animals) Nine pigs (Male, 5 ~ 6 weeks old, average weight 10 ~ 15 kg) were purchased from Hanabi Co., Ltd., and the experiment was applied after refining in an animal room for 2 weeks. The environment for breeding was applied to the environment of animal breeding room set at the temperature of 22 ± 3 ℃, relative humidity of 55 ± 5%, ventilation frequency of 15 times / hour, illumination of 150 ~ 300 Lux and illumination cycle of 12 hours.

(Methods and Dosage of Disclosed Substances) The method and dose were as shown in Table 4 below.

Test group Method of administration and dosage Dose Number of animals Test Approximately 1 1 mL of this agent per 16 kg of pig weight
Intramuscular injection once a day As Marbofloxacin
10 mg / kg body weight 3 two Test 2 1 mL of this agent per 25 kg of pig weight
Intramuscular injection once a day As Marbofloxacin
10 mg / kg body weight 3 two Test Approx. 3 1 mL of this agent per 30 kg of pig weight
Intramuscular injection once a day As Marbofloxacin
10 mg / kg body weight 3 two

Blood was collected from the jugular vein of pigs at 1 h, 3 h, 6 h, 12 h, 24 h, 2 days, 3 days, 5 days, 7 days and 10 days after administration of test substance mL, and then solidified at room temperature. The coagulated blood was centrifuged at 3000 rpm for 10 minutes to separate the serum and stored at -80 ° C until analysis.

(Quantitative Bioassay Test) Quantitative Bioassay was performed to determine the concentration of Marbofloxacin in serum. On the day of the test, the stored serum was dissolved at room temperature, and E. coli MIC was performed using ATCC 25922.

(Serum dilution) 0.05 mL of cation-adjusted MH medium was dispensed into all wells except for the first well of the prepared microplate. After 0.1 mL of serum was dispensed into the first well, 0.05 mL of the broth was mixed with broth that had been dispensed in the next well and diluted 2-fold in stair-step, diluted 1/1024 times.

(Strain dilution) E. coli ATCC 25922 was cultured on a 5% sheep blood agar and cultured in a 5% CO ₂ incubator at 37 ° C for 18-24 hours. Four to six single colonies were inoculated into cation-adjusted MH medium and cultured in a 37 ° C shaker incubator (180 rpm) for 2 to 6 hours. The turbidity of the cultured bacteria was determined by the method of MacFarland No. &lt; RTI ID = 0.0 &gt; 0.5 and adjusted to about 1.5 10 ⁸ CFU / mL, and diluted 100 times with the medium to prepare an inoculum of about 1.5 10 ⁶ CFU / mL.

(Inoculated and cultured) 0.05 mL of the bacterial solution adjusted to about 1.510 ⁶ CFU / mL in each well diluted with 2-fold of the serum, and cultured at 37 ° C. for 20 to 24 hours in an incubator, Respectively.

Pharmacokinetic analysis Pharmacokinetic analysis was analyzed using the program of BA-Calc 2007 1.0.0 (Food and Drug Administration).

(MIC and MBC) The MIC test was conducted in accordance with Clinical and Laboratory Standards Institute Document, M31-A3 (2008).

(Published strain) The bacteria used in the MIC test of this experiment are shown in Table 5.

Strain name Separation source Actinobacillus pleuropneumoniae pig Pasteurella multocida pig Haemophilus parasuis pig E. coli pig

(Preparation of Test Substance and Disclosed Concentration) 1 mL of each test substance is mixed with 9 mL of 95% EtOH (0.22 μm filtered) to prepare stock solution, and diluted with sterilized distilled water to 1 mg / mL of marbofloxacin effective ingredient Respectively. The published concentrations of the test substances are shown in Table 6.

Test substance menstruum Diluting solvent Published strain badge Published concentration range
(? μg / mL) Test Approximately 1
Test 2
Test Approx. 3 95% EtOH Sterilization
Distilled water A. pleuropneumoniae BHI ^a 0.001 to 3.75 ^c P. multocida CAMHB ^b 0.001 to 3.75 ^c H. parasuis BHI ^a 0.001 to 3.75 ^c E. coli CAMHB 0.001 to 3.75 ^c

^a: BHI containing 0.01% NAD, ^b: Cation-adjusted Muller Hiton broth, ^c : Marbolfoxacin concentration

Each bacterial strain was plated on an appropriate solid medium and cultured in a 5% CO ₂ incubator at 37 ° C for 18-24 hours. Four to six single colonies were then inoculated into the liquid medium shown in Table 7 below, , 5% CO ₂ incubator or 37 ° C shaking incubator (150 rpm) for 2 to 6 hours. The turbidity of the cultured bacteria was determined by the method of MacFarland No. &lt; RTI ID = 0.0 &gt; 0.5 and adjusted to about 1.5 10 ⁸ CFU / mL, and diluted 100 times with the medium to prepare an inoculum of about 1.5 10 ⁶ CFU / mL.

Strain name Culture medium Culture conditions Solid medium Liquid medium A. pleuropneumoniae Cocholate agar BHI ^a 37 ° C, 5% CO ₂ P. multocida 5% sheep blood agar CAMHB ^b 37 ℃ H. parasuis Cocholate agar BHI ^a 37 ° C, 5% CO ₂ E. coli 5% sheep blood agar CAMHB 37 ℃

^a : Brain heart infusion broth, ^b : cation-adjusted MH broth

(Bacterium inoculation and culture) A sterile 96-well microplate (F bottom, SPL) was prepared in advance and the name of the strain was written. Eight repeated wells were used for each test substance concentration group. In other words, 0.05 mL of each medium was dispensed into the prepared microplate. 0.05 mL of the test substance was mixed with 0.05 mL of the test substance in the wells of the highest concentration group, and 0.05 mL of the test substance was diluted with the broth and the azeotropy 2 in the wells of the next concentration group. . 0.05 mL of the bacterial solution adjusted to 1.510 ⁶ CFU / mL was dispensed into each well and cultured at 37 ° C in a 5% CO ₂ incubator or at 37 ° C for 18-24 hours.

(MIC [Minimum inhibitory concentration]). After 18-24 hours, the growth of the bacteria at the concentration was visually observed.

(MBC [Minimum bactericidal concentration] determination) After the MIC test, 0.05 mL was taken from the wells that did not grow germs, and the cells were streaked onto a chocolate agar containing no antibiotic and incubated at 37 ° C in a 5% CO ₂ incubator And cultured for 18-24 hours. MBC concentration was determined as 99.9% reduction in bacterial count compared to the inoculated bacteria. The MIC value of E. coli ATCC 25922 used as the QC of the MIC test for Marbolfoxacin was 0.0156 μg / mL, which was in accordance with the CLSI criteria (MIC range: 0.008-0.03 μg / mL). All MIC results were therefore considered valid.

<Experimental Results>

The pigs from each experimental group were administered the veterinary compositions 1 to 3 above and their doses are shown in Table 8 below.

Test group Average weight (kg) Average Dose (mL) Uniqueness during vaccination Test Approximately 1 18.3 ± 0.5 1.14 + 0.03 Injection was successful and there was no objection to injection. Test 2 18.4 ± 1.1 0.74 + 0.04 Relatively difficult to inject but no objection Test Approx. 3 18.2 ± 1.3 0.61 + 0.04 Injection was difficult, objection was reversed, and injection site hardness was observed for 2 days

(Quantitative bioassay validation and detection concentration measurement results) E. coli The concentration of marbofloxacin MIC for ATCC 25922 was 0.0156 μg / mL. (CLSI standard: 0.008 to 0.03 μg / mL).

(Results of analysis of blood marbofloxacin concentration) As a result of MIC test with isolated porcine serum collected over time, the concentration of Marbofloxacin in blood was confirmed as shown in Table 9 and FIG.

product name individual
number Blood concentration (μg / mL) 1h 6h 12h 24h 48h 72h 120h 168h 240h Test Approximately 1 One 0.998 3.994 1.997 1.997 0.499 0.250 0.062 - - 2 0.499 1.997 1.997 1.997 0.998 0.125 0.031 - - 3 0.998 1.997 1.997 1.997 0.499 0.125 0.031 - - Mean 0.832 2.662 1.997 1.997 0.666 0.166 0.042 - - SD 0.288 1.153 0.000 0.000 0.288 0.072 0.018 - - Test 2 4 0.998 1.997 1.997 1.997 0.499 0.250 0.031 - - 5 0.998 1.997 1.997 1.997 0.499 0.125 0.031 - - 6 0.499 0.998 1.997 1.997 0.499 0.125 0.031 - - Mean 0.832 1.664 1.997 1.997 0.499 0.166 0.031 - - SD 0.288 0.576 0.000 0.000 0.000 0.072 0.000 - - Test Approx. 3 7 0.998 1.997 1.997 0.998 0.499 0.125 - - - 8 0.499 0.998 0.998 0.998 0.499 0.125 - - - 9 0.499 3.994 3.994 3.994 0.499 0.125 - - - Mean 0.666 2.330 2.330 1.997 0.499 0.125 - - - SD 0.288 1.525 1.525 1.729 0.000 0.000 - - -

-: Not detected (lower than 0.0156 μg / mL)

Referring to the results of FIG. 4 and Table 9 above, the highest blood concentrations of test 1, test 2 and test 3 were 2.662, 1.997, and 2.330 μg / mL, respectively, at 6 hours or 12 hours after administration . Tests 1 and 2 showed marbofloxacin levels of 0.042 and 0.031 μg / mL in the blood until day 5 after administration, whereas testbone 3 was found to be marbofloxacin at 0.125 μg / mL until day 3 after administration.

(The results of comparative pharmacokinetic analysis of each drug, see Table 10) The highest blood level of test 1 (160 mg of marbophloxacin per 1 mL of injection) was observed at 6.0 ± 0.0 hours after administration, and the marbofloxacin concentration was 2.662 ± 1.153 μg / mL, and the half-life was 19.0 ± 4.3 h. The blood volume (AUC _{( inf )} ) was 94.016 ± 8.227 μg · h / mL.

Test drug 2 (Injection 1 mL per mabo floc reaper 250 mg) after administration was reached peak serum concentration of 1.997 ± 0.000 μg / mL to 8.0 ± 3.5 hours, the half-life was 16.6 ± 0.1 h, volume blood distribution (AUC _{( inf )} was found to be 85.030 ± 5.781 μg · h / mL.

The highest blood level of test compound 3 (300 mg of magoblocracin per injection) was 2.330 ± 1.525 μg / mL, 6.0 ± 0.0 hours after administration, and the half-life was 14.0 ± 3.9 h. The AUC (inf)) were 85.197 ± 52.205 μg · h / mL, respectively.

Pharmacokinetic parameter ^a unit Test Approximately 1
(MB 160 mg / mL) Test 2
(MB 250 mg / mL) Test Approx. 3
(MB 300 mg / mL) Lambda z h ^-1 4.333 + 1.155 4.667 ± 0.577 3.667 ± 1.155 AUC _(last) μg / mL 94.016 ± 8.227 84.282 ± 5.786 85.197 ± 52.205 AUC _{( inf )} μg / mL 95.233 + - 8.868 85.030 ± 5.781 87.725 ± 51.527 C _max μg / mL 2.662 ± 1.153 1.997 ± 0.000 2.330 1.525 T _max h 6.0 ± 0.0 8.0 ± 3.5 6.0 ± 0.0 t _1/2 h 19.0 ± 4.3 16.6 ± 0.1 14.0 ± 3.9 CL _{( inf )} / F L / h / kg 0.106 0.010 0.118 ± 0.008 0.139 + 0.066 Vz _(terminal) / F L / kg 2.870 + 0.446 2.830 + - 0.213 3.042 ± 1.883

Lambda z = elimination rate constant, AUC last = area under the serum concentration-time curve from time zero to time of last measurable concentration, AUC inf = area under the serum concentration-time curve from time zero to infinity, C max = peak concentration , T _max = time to peak concentration, t _1/2 = elimination half-life, CL _inf / F = Apparent total clearance of the drug from serum after extravascular administration, Vz _(terminal) after extravascular administration.

(MIC and MBC test results) The results of MIC and MBC of marbrochloxacin on porcine respiratory and gastrointestinal disease causative agents are shown in Table 11 below (MIC distribution pattern of the Dopyridazine DDS preparation on pig respiratory and digestive disease agents) and Table 12 And the concentration of MBC in the DDS preparation for the digestive tract disease agent).

Bacterium Number of isolates with MIC of (μg / mL) MIC ₅₀ MIC ₉₀ 0.015 0.023 0.029 0.047 0.059 0.094 0.117 0.188 0.234 0.357 A. pleruopneumoniae
(n = 4) - - - - - - One 3 - - 0.188 0.188 P. multocida (n = 4) One 3 - - - - - - - - 0.023 0.023 H. parasuis (n = 3) - - - - - - - 3 - - 0.188 0.188 E. coli (n = 4) - 4 - - - - - - - - 0.023 0.023

Bacterium Early
Number of inoculated bacteria (CFU / mL) MIC
(μg / mL) MBC
(μg / mL) Number of bacteria at MBC concentration
CFU / mL Bacterial clearance
(%) A. plueropneumoniae  (n = 4) 4.7 × 10 ⁵ 0.117 0.188 114 99.97 0.188 0.234 0 100 0.188 0.234 0 100 0.188 0.188 0 100 P. multocida
(n = 4) 2.3 × 10 ⁵ 0.023 0.029 0 100 0.015 0.023 2 99.998 0.023 0.029 0 100 0.015 0.015 2 99.998 H. parasuis
(n = 3) 1.0 × 10 ⁵ 0.188 0.188 0 100 0.188 0.188 2 99.998 0.188 0.188 0 100 E. coli
(n = 4) 1.0 × 10 ⁵ 0.023 0.023 0 100 0.023 0.023 26 99.97 0.023 0.029 0 100 0.015 0.023 0 100

(Pharmacokinetics / pharmacodynamics, PK / PD analysis)

One) Test drug  1 1 mL  medium Magoblockocaine 160 mg )

The results of PK / PD analysis for Test 1 are shown in Table 13 and FIG.

PK-PD integration * A. pleuropneumoniae P. multocida H. parasuis E. coli MIC: 0.188 [mu] g / mL MIC: 0.023 μg / mL MIC: 0.188 [mu] g / mL MIC: 0.023 μg / mL MBC: 0.234 占 퐂 / mL MBC: 0.029 μg / mL MBC: 0.188 占 퐂 / mL MBC: 0.029 μg / mL C _max / MIC ₉₀ 14.16 ± 6.13 115.77 ± 50.13 14.16 ± 6.13 115.77 ± 50.13 C _max / MBC 11.38 + - 4.93 91.82 + - 39.76 14.16 ± 6.13 91.82 + - 39.76

_0-/MIC (h)AUC

_0- / MIC (h) 550.52 ± 47.17 4140.57 + - 385.57 550.52 ± 47.17 4140.57 + - 385.57 AUC

_0- / MBC (h) 406.98 +/- 37.90 3283.90 + - 305.80 550.52 ± 47.17 3283.90 + - 305.80 T > MIC (h) 71.66 ± 13.5 138.48 ± 10.21 71.66 ± 13.5 138.48 ± 10.21 T > MBC (h) 63.89 + - 9.98 130.88 ± 12.79 71.66 ± 13.5 130.88 ± 12.79

_{* C max / MIC = ratio of} in vivo C max to MIC, C max / MBC = ratio of in vivo C max to MBC, AUC / MIC = ratio of in vivo AUC to MIC, AUC / MBC = ratio of in vivo AUC to MBC, T> MIC = total time that marbofloxacin concentration exceeded MIC, T> MBC = total time that marbofloxacin concentration exceeded MBC

Referring to FIG. 5 and Table 13, the test marbofloxacin concentration in the blood after a single dose of about 1 to a pig muscles pleuropneumeniae A., P. multocida, H. parasius, E. coli maintained above the MIC ₉₀ of the bacteria (C _max ) was 14.16 ± 6.13 times, 115.77 ± 50.13 times, and 14.16 times higher than the MIC ₉₀ of each bacterium, respectively. The maximum blood concentration (C _max ) was 71.66 ± 13.51 h, 138.48 ± 10.21 h, 71.66 ± 13.5 h and 138.48 ± 10.21 h, ± 6.13 times and 115.77 ± 50.13 times, respectively. The maximum duration of blood marbofloxacin was 63.89 ± 9.98 h, 130.88 ± 12.79 h, 71.66 ± 13.5 h and 130.88 ± 12.79 h, respectively, and the highest blood concentration (C _max ) 4.93 times, 91.82 ± 39.76 times, 14.16 ± 6.13 times and 91.82 ± 39.76 times, respectively.

2) Test drug  2 1 mL  medium Magoblockocaine 250 mg )

The results of PK / PD analysis for test 2 are shown in Table 14 and FIG. 6 below.

PK-PD integration * A. pleuropneumoniae P. multocida H. parasuis E. coli MIC: 0.188 [mu] g / mL MIC: 0.023 μg / mL MIC: 0.188 [mu] g / mL MIC: 0.023 μg / mL MBC: 0.234 占 퐂 / mL MBC: 0.029 μg / mL MBC: 0.188 占 퐂 / mL MBC: 0.029 μg / mL C _max / MIC ₉₀ 10.62 ± 0.00 86.83 ± 0.00 10.62 ± 0.00 86.83 ± 0.00 C _max / MBC 8.53 ± 0.00 68.86 ± 0.00 10.62 ± 0.00 68.86 ± 0.00

_0-/MIC (h)AUC

_0- / MIC (h) 452.29 ± 30.75 3696.94 + - 251.37 452.29 ± 30.75 3696.94 + - 251.37 AUC

_0- / MBC (h) 363.38 ± 24.71 2932.06 ± 199.36 452.29 ± 30.75 2932.06 ± 199.36 T > MIC (h) 70.47 ± 12.64 132.54 + 0.21 70.47 ± 12.64 132.54 + 0.21 T > MBC (h) 62.87 + - 10.28 123.27 ± 0.15 70.47 ± 12.64 123.27 ± 0.15

_{* C max / MIC = ratio of} in vivo C max to MIC, C max / MBC = ratio of in vivo C max to MBC, AUC / MIC = ratio of in vivo AUC to MIC, AUC / MBC = ratio of in vivo AUC to MBC, T> MIC = total time that marbofloxacin concentration exceeded MIC, T> MBC = total time that marbofloxacin concentration exceeded MBC

Referring to Table 14, and 6 above, the test marbofloxacin concentration in the blood after a single dose of about 2 in the pig muscles pleuropneumeniae A., P. multocida, H. parasius, the E. coli bacteria above the MIC ₉₀ The maximum blood concentration (C _max ) was 10.62 ± 0.00 times, 86.83 ± 0.00 times higher than the MIC ₉₀ of each bacterium, 10.62 ± 0.00 times and 86.83 ± 0.00 times higher than the control group. The maximum duration of blood marbofloxacin concentration was 62.87 ± 10.28 h, 123.27 ± 0.15 h, 70.47 ± 12.64 h and 123.27 ± 0.15 h, respectively, and the highest blood concentration (C _max ) 0.008 times, 68.86 ± 0.00 times, 10.62 ± 0.00 times, and 68.86 ± 0.00 times, respectively.

3) Test drug  3 1 mL  medium Magoblockocaine 300 mg )

The results of PK / PD analysis for test 3 are shown in Table 15 and FIG. 7 below.

PK-PD integration * A. pleuropneumoniae P. multocida H. parasuis E. coli MIC: 0.188 [mu] g / mL MIC: 0.023 μg / mL MIC: 0.188 [mu] g / mL MIC: 0.023 μg / mL MBC: 0.234 占 퐂 / mL MBC: 0.029 μg / mL MBC: 0.188 占 퐂 / mL MBC: 0.029 μg / mL C _max / MIC ₉₀ 12.39 8.11 101.29 + - 66.32 12.39 8.11 101.29 + - 66.32 C _max / MBC 9.96 + - 6.52 80.33 + - 52.60 12.39 8.11 80.33 + - 52.60

_0-/MIC (h)AUC

_0- / MIC (h) 466.62 + - 274.08 3814.15 + - 2240.31 466.62 + - 274.08 3814.15 + - 2240.31 AUC

_0- / MBC (h) 374.90 ± 220.20 3025.01 占 1776.80 466.62 + - 274.08 3025.01 占 1776.80 T > MIC (h) 63.66 ± 1.13 111.05 + 0.07 63.66 ± 1.13 111.05 + 0.07 T > MBC (h) 56.65 ± 0.06 108.78 ± 0.05 63.66 ± 1.13 108.78 ± 0.05

_{* C max / MIC = ratio of} in vivo C max to MIC, C max / MBC = ratio of in vivo C max to MBC, AUC / MIC = ratio of in vivo AUC to MIC, AUC / MBC = ratio of in vivo AUC to MBC, T> MIC = total time that marbofloxacin concentration exceeded MIC, T> MBC = total time that marbofloxacin concentration exceeded MBC

Referring to Table 15 and Figure 7, the test marbofloxacin concentration in the blood after about 3 in one administration to the pig muscles pleuropneumeniae A., P. multocida, H. parasius, E. coli maintained above the MIC ₉₀ of the bacteria (C _max ) was 12.39 ± 8.11 times, 101.29 ± 66.32 times, and 12.39 ± 12.39 times higher than the MIC ₉₀ of each bacterium, respectively. ± 8.11 times and 101.29 ± 66.32 times, respectively. Time at which the blood marbofloxacin concentration maintained higher than the MBC for each bacteria 56.65 ± 0.06 h, 108.78 ± 0.05 h, 63.66 ± 1.13 h and 108.78 were ± 0.05 h, peak serum concentration (C _max) is 9.96 than the MIC ₉₀ of each bacterial ± 6.52 times, 80.33 ± 52.60 times, 12.39 ± 8.11 times, and 80.33 ± 52.60 times, respectively

Above, peak blood concentrations were reached from 6 hours to 12 hours after intramuscular injection of test 1 (MB 160 mg / mL), test 2 (MB 250 mg / mL) and test 3 (MB 300 mg / mL) The serum levels of marbofloxacin were 2.662 ± 1.153, 1.997 ± 0.000 and 2.330 ± 1.525 μg / mL, respectively. Marbofloxacin was detected as 0.042 and 1.052 μg / mL in the blood until the 5th day after the administration of test 1 (MB 160 mg / mL) and test 2 (MB 250 mg / mL). However, the test substance 3 (MB 300 mg / mL) was found to have a concentration of 0.125 μg / mL of marbofloxacin on the third day after administration. These results suggest that the test drug 3 (MB 300 mg / mL) is highly viscous and is not absorbed properly.

The plasma half-lives of test 1 (MB 160 mg / mL) and test 2 (MB 250 mg / mL) were 19.0 ± 4.3 h and 16.6 ± 0.1 h, respectively, ). (AUC _{( inf )} ) of test 1, test 2 and test 3 were 95.233 ± 8.868, 85.030 ± 5.781 and 87.725 ± 51.527 μg / mL, respectively, Respectively.

The highest concentrations of marbofloxacin in the blood of test 1 (MB 160 mg / mL), test 2 (MB 250 mg / mL) and test 3 (MB 300 mg / mL) were 2.663 ± 1.153, 1.997 ± 0.000, 2.313 ± 1.525 μg / mL, respectively, which were 14.16 ± 6.13 times, 10.62 ± 0.00 times, and 12.39 ± 8.11 times higher than the MIC ₉₀ of A. pleuropneumoniae , respectively, and the time to maintain higher than MIC ₉₀ was 71.66 ± 13.51 h, 70.47 ± 12.64 h and 63.66 占 1.13 h. In addition, A. 11.38 ± 4.93 times, 8.53 ± 0.00 times, and 9.96 ± 6.52 times higher than MBC of pleuropneumoniae , respectively. The time to maintain higher than MBC was 63.89 ± 9.98 h, 62.87 ± 10.28 h and 56.65 ± 0.06 h, respectively.

The highest concentration of marbofloxacin in blood of test 1 (MB 160 mg / mL), test 2 (MB 250 mg / mL) and test drug 3 (MB 300 mg / mL) was higher than that of P. multocida and E. coli MIC ₉₀ 115.77 ± 50.13 times, 86.83 ± 0.00 times, and 101.29 ± 66.32 times, respectively, and the concentrations were higher than MIC ₉₀ for 138.48 ± 10.21 h, 132.54 ± 0.21 h and 111.05 ± 0.07 h, respectively. In addition, the highest blood concentrations of each test substance were 91.82 ± 39.76, 68.86 ± 0.00, and 80.33 ± 52.60 times higher than those of MBC, respectively. h.

Test drug 1 (MB 160 mg / mL) , test product 2 (MB 250 mg / mL) and test product 3 (MB 300 mg / mL) peak serum concentration of 14.16 ± 6.13 respectively, than the MIC ₉₀ and MBC of H. parasuis in And 10.62 ± 0.00 times 12.39 ± 8.11 times higher than those of MIC ₉₀ and MBC, respectively. The time to stay higher than MIC ₉₀ and MBC was 71.66 ± 13.5 h, 70.47 ± 12.64 h and 63.66 ± 1.13 h, respectively.

These results suggest that drug 1 (MB 160 mg / mL) and test drug 2 (MB 250 mg / mL) have similar drug persistence.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (12)

Preparing a peroxide oil for preparing a mixture for supporting antibiotics by mixing the peroxide oil, which is an oil containing an unsaturated fatty acid oxidized so that the peroxide value is not less than 300 meq / kg, with the stabilized oil containing the saturated oil; And
A mixture of the mixture for supporting antibiotics and an antibiotic component containing a carboxyl group, an amine group or both in a molecule, and a sustained-release form of an antibiotic containing the sustained-release antibiotic-supporting oil in which the antibiotic component is chemically bonded to the peroxide oil A method for the preparation of a sustained-release veterinary composition for the prevention or treatment of a disease caused by bacteria in an animal, comprising the step of preparing a preparation. The method according to claim 1,
Wherein the peroxide oil preparation step comprises: an oxidation step of oxidizing an oil containing an unsaturated fatty acid to produce a peroxide oil having a peroxide value of 300 meq / kg or more; And a stabilizing step of mixing a stabilizing oil containing a saturated oil with the peroxide oil to prepare a mixture for supporting antibiotics. 3. The method of claim 2,
Wherein the oil containing the unsaturated fatty acid has a polyunsaturated fatty acid content of at least 50% by weight, based on the total oil, of the composition. 3. The method of claim 2,
Wherein the oxidation step is carried out at an oxidation temperature of 70 to &lt; RTI ID = 0.0 &gt; 120 C. &lt; / RTI &gt; The method according to claim 1,
Wherein the antibiotic component comprises any one selected from the group consisting of marbophloxacin, danofloxacin, enrofloxacin, their respective veterinarily acceptable salts, and combinations thereof as an active ingredient. Wherein the method comprises the steps of: 3. The method of claim 2,
Wherein said stabilizing oil is applied in an amount of 50 to 99% by volume based on the sum of said peroxide oil and said stabilizing oil. The method according to claim 1,
Wherein the content of the antibiotic component contained in the sustained-release antibiotic-supporting oil is less than 300 mg / mL, for the prevention or treatment of a disease caused by a bacterium. Peroxide oil having a peroxide value of 300 meq / kg or more; A stabilizing oil containing a saturated oil; And an antibiotic active ingredient comprising a carboxyl group, an amine group, or both in the molecule. The composition for the preparation of a sustained release preparation for the prevention or treatment of a disease caused by an animal bacterium. Prevention of diseases caused by bacteria in an animal, comprising a sustained release formulation of antibiotics comprising a stabilized oil containing a sustained-release antibiotic-bearing oil and an antibiotic-bearing oil chemically bonded to a chain of fatty acids, A sustained-release veterinary composition for treatment. 10. The method of claim 9,
The sustained-release veterinary composition for preventing or treating a disease caused by a bacterium, wherein the sustained-release preparation of the antibiotic contains the antibiotic component at 300 mg / mL or lower and has a viscosity of 280 cP (20 캜) or lower. 10. The method of claim 9,
The sustained-release veterinary composition for the prevention or treatment of a disease caused by a bacterium, wherein the sustained-release preparation of the antibiotic is a injectable preparation and has a drug persistence of 60 hours or more after the injection. A method of treating an animal other than a human, comprising the step of injecting a sustained veterinary composition for the prevention or treatment of a disease caused by the bacterium according to claim 1.

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