CN110898061B

CN110898061B - An antibacterial agent suitable for oral administration to animals

Info

Publication number: CN110898061B
Application number: CN201911195283.9A
Authority: CN
Inventors: 王永东; 操基元; 祝诗发; 黄志鹏; 程稳
Original assignee: Guang Zhou Yuan Tu Biological Ang Chemical Technology Co ltd; GENIFARM (GUANGZHOU) TECHNOLOGY Inc
Current assignee: Guang Zhou Yuan Tu Biological Ang Chemical Technology Co ltd; GENIFARM (GUANGZHOU) TECHNOLOGY Inc
Priority date: 2018-12-19
Filing date: 2019-11-28
Publication date: 2022-04-15
Anticipated expiration: 2039-11-28
Also published as: CN110898061A; CN114767679A

Abstract

The invention discloses an antibacterial agent suitable for oral administration to animals, which comprises quinolone medicaments or pharmaceutically acceptable salts thereof and tannin or pharmaceutically acceptable salts thereof. The invention also discloses a preparation method and application of the antibacterial product, and a pharmaceutical preparation containing the antibacterial product. The research of the invention finds that the combination of the quinolone medicaments and the tannin eliminates the unpleasant odor and taste of the quinolone medicaments, and solves the problem that the odor and taste of the existing quinolone medicaments bring food refusal and drink refusal to animals sensitive to smell and/or taste during administration; meanwhile, pharmacodynamic test results show that the product has an obvious synergistic bacteriostatic action on sensitive bacteria, can obviously reduce the minimum bacteriostatic concentration of quinolone drugs, reduces the use amount of the quinolone drugs, and has a good application prospect.

Description

An antibacterial agent suitable for oral administration to animals

Technical Field

The invention belongs to the technical field of medicines. And more particularly to an antibacterial agent suitable for oral administration to animals.

Background

Various domestic animals, including horses, cattle, sheep, pigs, etc., have a sensitive set of sensory organs (smell, taste, hearing, vision). All domestic animals rely on these sense organs to adapt to the external complex environmental conditions to obtain survival and breed offspring. The livestock can distinguish palatability of feed and select drinking water by using sense of smell and taste (instinct of sense of smell of livestock and its application in animal husbandry, Luweimin, zoology of livestock, 1 st 1989). The vision of the pigs in the domestic animals is poor, the sight distance is short, the range is small, and therefore the identification and judgment of the pigs on things mainly depends on the sense of smell and the sense of hearing. The pig has long nose tube and wide smelling area, the total area of smelling mucosa villi is large, the olfactory nerves with well-developed distribution are distributed, and the recognition capability of the smell is about 7-8 times larger than that of a human (the application of the smell of the pig in pig raising production, Tanyuan, Guangdong animal husbandry veterinary science and technology, volume 19, No. 4 in 1994, P35). Pregnant sows, lactating sows and breeding boars are especially sensitive to smell for ensuring the reproduction of offspring due to their biological instincts. Good feeding behavior is the basis for maintaining growth and development of animal organism and improving production performance, and smell sense and taste sense are two most important sense senses in animal feeding process. The food intake of animals is closely related to the sense of smell and taste, and the sense of smell is the sense of the animal body generated by receiving the volatile substances in the long distance of the food. The smell sense can judge the preference of the animal body to food and make a decision on whether to eat or not by only the smell of the animal body without contacting the food. In the actual animal husbandry production, the sense of smell and taste are very important for animals with self-feeding ability, animals with blindness, free-range animals and livestock, and the survival rate, the body health, the breeding and the like of the animals are directly determined. For large-scale cultivation, the optimal mode of group administration is oral administration, namely administration by mixed drinking or mixed feeding, and the oral administration has the characteristics of reducing animal stress, reducing labor intensity, quickly preventing and treating group animal diseases and the like.

Quinolone (Quinolone) is also called pyrrone or pyridone acid, and is an artificially synthesized antibacterial drug containing a Quinolone structure. Quinolones target bacterial deoxyribonucleic acid (DNA), interfering with DNA gyrase, further causing irreversible damage to chromosomes, and preventing bacterial cells from dividing. The medicine is not affected by plasmid conducted drug resistance, so that the medicine has no cross drug resistance with many antibacterial drugs, has the characteristics of wide antibacterial spectrum, strong antibacterial activity, no cross drug resistance with other antibacterial drugs, small toxic and side effects and the like, is widely applied to livestock breeding, aquatic product breeding and other breeding industries, and is used for disease control in breeding of chicken, ducks, geese, pigs, cows, sheep, fish, shrimps, crabs and the like.

However, quinolone drugs have an unpleasant odor and taste. The odor and taste can cause an aversive response in animals, in which case, olfactory and gustatory sensitive animals can produce food and drink refusals, and thus quinolone drugs are difficult to apply to the treatment of bacterial infections in such animals by oral administration. The choice of food by the animal through smell and taste is the basis for achieving self-protection. This behaviour is considered to be a defense mechanism against toxic substances, which plays a crucial role during the long-term evolution of animals (HernessMS, gilbertson t.a. annurevphysiol,1999,61: 873-. The unpleasant odor and taste of the quinolone drugs are considered to be harmful to animal organisms by olfactory or gustatory sensitive animals, so that the feed intake or water intake of the quinolone drugs can be rapidly reduced by simple mixed feeding or mixed drinking administration, even complete food refusal and water refusal are avoided, the intake of the quinolone drugs is influenced, and the purpose of treating the animals suffering from bacterial or mycoplasma infection cannot be achieved by oral administration.

To overcome the animal's problem of food refusal, korean patent application No. 20020089783 discloses inclusion of enrofloxacin by β -cyclodextrin, addition of taste masking agents such as crispy rice, aspartame, etc.; chinese patent No. 200610049243.X adopts gelatin encapsulation technology to mask taste and prepare the medicinal additive; chinese patent No. 200510050039.5 discloses an enrofloxacin microcapsule and a preparation method thereof, wherein enrofloxacin microcapsule is prepared by coating material, so as to achieve the purpose of taste masking; chinese patent application No. 200810091851.6 discloses a method for improving the palatability of enrofloxacin by an envelope microcapsule technique. However, after the patent products are added into the feed, when the patent products are in contact with water or are broken when the patent products are in contact with saliva or chewed in the oral cavity of an animal, the unpleasant taste of the main drug is easy to release, so that the ingestion of the animal is reduced, even the animal refuses to eat, and the purpose of oral administration is difficult to achieve.

Disclosure of Invention

The invention aims to solve the technical problem that the existing quinolone drugs give unpleasant feelings to animals due to the odor and taste of the drugs when the drugs are taken, so that the animals refuse to eat and drink. According to the invention, researches show that the unpleasant odor and taste of the quinolone medicaments can be eliminated by using the quinolone medicaments and the tannin in a combined manner, so that the problems are solved, the minimum antibacterial concentration of the quinolone medicaments can be reduced, and the using amount of the quinolone medicaments is reduced.

It is an object of the present invention to provide an antibacterial agent suitable for oral administration to an animal.

The above purpose of the invention is realized by the following technical scheme:

the research of the invention finds that the carbostyril drug and the tannin have strong bitter taste and strong astringent taste, and the combined use of the carbostyril drug and the tannin can avoid unpleasant feeling when the carbostyril drug and the tannin are used separately, and simultaneously can play a remarkable synergistic effect, reduce the minimum inhibitory concentration of the carbostyril drug and reduce the use amount of the carbostyril drug, thereby providing a new animal antibacterial product.

An antimicrobial product suitable for oral administration to an animal comprising the following components:

(i) quinolones (Quinolones) or pharmaceutically acceptable salts thereof;

(ii) tannins (Tannins) or pharmaceutically acceptable salts thereof;

wherein, the quinolone refers to a compound containing a quinolone structure, and the compound has the following structure:

R¹and R⁵The dotted line between represents R¹And R⁵Are independent groups or they are linked to form a ring;

R¹represents alkyl, cycloalkyl or phenyl;

R²represents a hydrogen atom or a halogen;

R³represents halogen;

R⁴represents a hydrogen atom, an alkyl group,

A five-membered heterocyclic group or a six-membered heterocyclic group;

R⁵represents a hydrogen atom, a halogen, a cyano group, an alkyl group, or an alkoxy group.

When R is⁵，R¹When the compounds are connected to form a ring, the compounds and carbon atoms and nitrogen atoms connected with the compounds form a six-membered heterocyclic ring, and the six-membered heterocyclic ring contains one or more N, O, S atoms;

one or more hydrogen atoms on the alkyl, cycloalkyl, alkoxy, five-membered heterocyclic group, six-membered heterocyclic group or phenyl can be substituted by a substituent which is independently selected from alkyl, alkoxy, acyl, halogen, cyano, amino, nitro, hydroxyl;

the term "alkyl" refers to a straight or branched chain saturated hydrocarbon group containing only carbon and hydrogen atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, including but not limited to methyl (-CH)₃) Ethyl (-CH)₂CH₃) Is rightPropyl (-CH)₂CH₂CH₃) Isopropyl group (-CH (CH)₃)₂) N-butyl (-CH)₂CH₂CH₂CH₃) Isobutyl (-CH)₂CH(CH₃)₂) Sec-butyl (-CH (CH)₃)CH₂CH₃) Tert-butyl (-C (CH)₃)₃) N-pentyl (-CH)₂CH₂CH₂CH₂CH₃) 2-pentyl (-CH (CH)₃)CH₂CH₂CH₃) 3-pentyl (-CH (CH)₂CH₃)₂) 2-methyl-2-butyl (-C (CH)₃)₂CH₂CH₃) 3-methyl-2-butyl (-CH (CH)₃)CH(CH₃)₂) 3-methyl-1-butyl (-CH)₂CH₂CH(CH₃)₂) 2-methyl-1-butyl (-CH)₂CH(CH₃)CH₂CH₃) N-hexyl (-CH)₂CH₂CH₂CH₂CH₂CH₃) 2-hexyl (-CH (CH)₃)CH₂CH₂CH₂CH₃) 3-hexyl (-CH (CH)₂CH₃)(CH₂CH₂CH₃) 2-methyl-2-pentyl (-C (CH))₃)₂CH₂CH₂CH₃) 3-methyl-2-pentyl (-CH (CH)₃)CH(CH₃)CH₂CH₃) 4-methyl-2-pentyl (-CH (CH)₃)CH₂CH(CH₃)₂) 3-methyl-3-pentyl (-C (CH)₃)(CH₂CH₃)₂) 2-methyl-3-pentyl (-CH (CH)₂CH₃)CH(CH₃)₂) 2, 3-dimethyl-2-butyl (-C (CH)₃)₂CH(CH₃)₂) 3, 3-dimethyl-2-butyl (-CH (CH)₃)C(CH₃)₃) N-heptyl, n-octyl, and the like.

The term "alkoxy" means an alkyl group attached to the rest of the molecule through an oxygen atom, wherein the alkyl group has the meaning as described herein;

the term "cycloalkyl" refers to a cyclic organic group containing only two atoms of carbon and hydrogen, preferably 3 to 10 carbon atoms, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, and the like;

the term "halogen" refers to fluorine, chlorine, bromine, iodine;

the term "five-membered heterocyclic group" means a group formed of a cyclic organic compound composed of carbon atoms and non-carbon atoms, the sum of the number of carbon atoms and the number of non-carbon atoms being 5. The five-membered heterocyclic group may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more, and may be independently selected from halogen, hydroxy, amino, alkoxy, alkyl, cyano, aryl, heterocyclic group.

The term "six-membered heterocyclic group" means a group formed of a cyclic organic compound composed of carbon atoms and non-carbon atoms, the sum of the number of carbon atoms and the number of non-carbon atoms being 6. The six-membered heterocyclic group may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more, and may be independently selected from halogen, hydroxy, amino, alkoxy, alkyl, cyano, aryl, heterocyclic group.

The term "aryl" refers to substituted or unsubstituted planar ring systems possessing conjugation, as is common, e.g., benzene rings, naphthalene rings, furans, pyrroles, indoles, thiophenes, imidazoles, pyrazoles, oxazoles, and the like.

One or more CH of the above-mentioned alkyl, cycloalkyl or alkoxy radicals₂A group may be replaced by an O or S atom, a CH ≡ CH-group and/or a C ≡ C-group, or wherein one or more hydrogen atoms of the alkyl, cycloalkyl or alkoxy groups may be replaced by halogen, hydroxy, carbonyl, substituted or unsubstituted aryl, C3-C10 cycloalkyl.

Preferably, the quinolone of the present invention is preferably a compound having the structure:

x is independently selected from carbon atom, nitrogen atom and oxygen atom; y is independently selected from carbon atoms and nitrogen atoms.

Wherein: r¹Represents alkyl, cyclopropyl or phenyl;

R²represents a hydrogen atom or a fluorine atom;

R⁴represents a hydrogen atom, a methyl group,

A five-membered heterocyclic group or a six-membered heterocyclic group;

R⁵represents a hydrogen atom, a fluorine atom, a cyano group, an alkyl group, or an alkoxy group.

One or more hydrogen atoms on the alkyl, cyclopropyl, alkoxy, five-membered heterocyclic group, six-membered heterocyclic group or phenyl can be substituted by a substituent which is independently selected from alkyl, alkoxy, acyl, halogen, cyano, amino, nitro and hydroxyl.

More preferably, the substituents in the quinolone are preferably as follows:

R¹is selected from C_1-3Alkyl, cyclopropyl or phenyl;

R²selected from a hydrogen atom or a fluorine atom;

R⁴selected from the following structures:

R⁵selected from hydrogen atom, fluorine atom, cyano group, C_1-3Alkyl radical, C_1-3An alkoxy group.

Said C is_1-3Alkyl, cyclopropyl, C_1-3One or more hydrogen atoms on the alkoxy group, the five-membered heterocyclic group, the six-membered heterocyclic group or the phenyl group may be substituted by a substituent independently selected from the group consisting of alkyl, alkoxy, acyl, halogen, cyano, amino, nitro, hydroxyl.

As a preferable mode, the quinolone is preferably Enrofloxacin Enrofloxacin, Marbofloxacin, Difloxacin Difloxacin, Sarafloxacin, Ciprofloxacin Ciprofloxacin, Danofloxacin, Flumequine, Benofloxacin, Norfloxacin, Ofloxacin, Lomefloxacin Lomefloxacin, Pefloxacin Pefloxacin, Orbifloxacin, Ibafloxacin, Pradofloxacin.

As a more preferable embodiment, the above quinolone is more preferably: enrofloxacin, Marbofloxacin, Difloxacin, Sarafloxacin, Ciprofloxacin, Danofloxacin, Flumequine, Benofloxacin, Norfloxacin, Ofloxacin Ofloxacin, Lomefloxacin, Pefloxacin.

As a more preferable mode, the above quinolone is more preferably: enrofloxacin, Marbofloxacin, Difloxacin, Sarafloxacin, Ciprofloxacin, Danofloxacin, Flumequine, Orbifloxacin.

In a preferred embodiment, the quinolone is Enrofloxacin.

In a preferred embodiment, the quinolone is Danofloxacin.

In a preferred embodiment, the quinolone is Sarafloxacin.

In a preferred embodiment, the quinolone is Orbifloxacin.

Preferably, in the antibacterial product, the weight ratio of the components (i) and (ii) is preferably 5: 95-95: 5.

As a preferable mode, the weight ratio of the components (i) and (ii) is more preferably 15:85 to 85: 15.

As a more preferred embodiment, the weight ratio of the components (i) and (ii) is more preferably 25:75 to 75: 25.

Preferably, the antibacterial product suitable for oral administration to an animal is prepared by uniformly mixing components (i) and (ii).

The preparation method of the antibacterial product for oral administration to animals is that (i) quinolone or pharmaceutically acceptable salt thereof and (ii) tannin or pharmaceutically acceptable salt thereof are uniformly mixed by a chemical method or a physical method.

As an alternative preferred solution, the antimicrobial product can also be prepared by the following steps:

(1) (ii) forming (i) the quinolone or a pharmaceutically acceptable salt thereof directly or into a solution or suspension;

(2) (iii) forming the (ii) tannin or a pharmaceutically acceptable salt thereof directly or as a solution or suspension; (ii) a

(3) And, at least one of quinolone or tannin is solution or suspension, then mix (1) and (2), filter, collect the precipitate;

(4) and (5) drying the precipitate to obtain an antibacterial product.

As another alternative, the antibacterial product may be prepared by directly and homogeneously mixing (i) the quinolone or a pharmaceutically acceptable salt thereof and (ii) the tannin or a pharmaceutically acceptable salt thereof.

The direct and uniform mixing in the present application refers to a mixing mode without the participation of a solvent, and specifically, the preparation of the antibacterial product can be realized by the following steps:

(1) weighing quinolone or pharmaceutically acceptable salt thereof;

(2) weighing tannin or pharmaceutically acceptable salt thereof;

(3) uniformly mixing quinolone or pharmaceutically acceptable salt thereof and tannin or pharmaceutically acceptable salt thereof to obtain the antibacterial product.

The uniform mixing is usually carried out by pulverizing the quinolone or the pharmaceutically acceptable salt thereof, the tannin or the pharmaceutically acceptable salt thereof, sieving, and then mixing.

After the uniform mixing, the antibacterial product can be better formed by a granulation method. The granulation is preferably wet granulation, and the granules are dried after granulation to obtain the antibacterial product.

The crushing and sieving can be completed by adopting crushing equipment and sieving equipment which are commonly used in the field, no special requirements are required on the process, and the crushing and sieving aims are both to realize more uniform mixing of the crushing equipment and the sieving equipment.

The quinolone or tannin can be formed into solution or suspension, and solvent, and optionally inorganic acid or organic acid can be added to increase its solubility.

The solvent refers to any solvent that can dissolve the quinolone and tannin, including but not limited to: water, methanol, ethanol, isopropanol, acetone, ethyl acetate, isopropyl acetate, methyl tert-butyl ether, acetonitrile, N-dimethylformamide, dichloromethane, trichloromethane, dichloroethane and mixtures of such solvents.

The inorganic or organic acid may be hydrochloric acid, sulfuric acid, phosphoric acid, formic acid, acetic acid, propionic acid, butyric acid, methanesulfonic acid, lactic acid, benzoic acid, fumaric acid, malic acid, arginine, glutamic acid, aspartic acid, tartaric acid, etc.

Based on the method, the invention provides an antibacterial product prepared by the method.

The antibacterial product can be used as a broad-spectrum antibacterial drug and can be used for preparing drugs for resisting animal bacterial infection. The method eliminates the unpleasant odor and taste of the quinolone drugs while reducing the dosage of the quinolone drugs, and solves the problem that the animal refuses to eat and drink due to the unpleasant feeling brought to the animal by the odor and taste of the drugs when the existing quinolone drugs are taken.

The animal is an olfactory and/or gustatory sensitive animal. The olfactory and/or gustatory sensitive animals include, but are not limited to, pigs, cattle, sheep, horses, dogs, cats. Preferably pig, cattle, dog.

The invention provides a pharmaceutical preparation, which comprises the antibacterial product and one or more pharmaceutically acceptable auxiliary materials.

The pharmaceutically acceptable auxiliary materials are selected from: at least one of solvents, solubilizers, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integration agents, permeation enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, antifoaming agents, thickeners, encapsulation agents, humectants, absorbents, diluents, flocculants and deflocculants, filter aids, release retardants, and combinations thereof.

The antibacterial product can also be prepared into suspension, dry suspension, granules, powder, premix, pills, tablets, effervescent agents, solution and the like.

The administration mode of the antibacterial product to animals can be drinking water administration or mixed feeding administration.

The invention has the following beneficial effects:

according to the antibacterial product, the quinolone medicaments and the tannin are combined, so that the prepared product is stable in finished product, good in dispersity, free of unpleasant odor and taste, and good in antibacterial effect and pharmacokinetic characteristic, has a remarkable synergistic effect, maintains the same antibacterial effect while reducing the using dose of the quinolone medicaments, and therefore reduces the risk of side effects or adverse reactions of antibiotics; meanwhile, the drug combination is slow-release and controlled-release, easy to absorb and good in treatment effect.

The antibacterial product for animals also enables animal groups to take medicines by a mixed feeding or drinking water administration method, reduces the labor intensity of individual administration, is convenient and quick, and avoids the problem of insufficient medicine intake caused by the decrease of sick appetite of animals.

Drawings

FIG. 1 is a thermogravimetric analysis of tannin.

FIG. 2 is a thermogravimetric analysis of ciprofloxacin.

FIG. 3 is a thermogravimetric analysis of the product of example 12.

FIG. 4 is a graph showing the effect of the product of example 12 on the growth of Staphylococcus aureus or Escherichia coli.

FIG. 5 shows the effect of products 31A, 31B, 31C, 31D of example 31 on the growth of E.coli.

Detailed Description

The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Unless otherwise indicated, reagents and raw auxiliary materials used in the following examples are commercially available.

Example 1

The enrofloxacin and the tannin are crushed, sieved and mixed according to the mass portion of 5:95 to prepare the antibacterial product for animals.

Example 2

The danofloxacin mesylate and the tannin are crushed, sieved and mixed according to the weight portion of 10:90, and then are granulated by a wet method and dried to prepare the antibacterial product for animals by a conventional method.

Example 3

The ciprofloxacin hydrochloride and the tannin are crushed, sieved and mixed according to the mass portion of 20:80, and the antibacterial product for animals is prepared by a conventional method.

Example 4

The marbofloxacin and the tannin are crushed, sieved and mixed according to the mass ratio of 30:70, and then are granulated by a wet method and dried to prepare the antibacterial product for animals by a conventional method.

Example 5

Putting 50 parts by weight of sarafloxacin in 1500 parts of water, stirring, and dropwise adding hydrochloric acid until the sarafloxacin is completely dissolved; dissolving 50 parts by weight of tannin in 500 parts by weight of water, slowly adding the tannin into the solution, stirring, reacting for 1 hour at 50 ℃, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 6

Dissolving 50 parts by weight of difloxacin hydrochloride in 2000 parts of water, dissolving 50 parts by weight of tannin in 500 parts of water, combining the two parts of solution, stirring uniformly, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 7

Putting 50 parts by weight of sarafloxacin into 2000 parts of water, dissolving 50 parts by weight of tannin into 500 parts of water, combining the sarafloxacin and the tannin, stirring uniformly, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 8

Dissolving 60 parts by weight of enrofloxacin in 2000 parts by weight of dichloromethane, dissolving 40 parts by weight of tannin in 800 parts by weight of water, combining the two parts of solution, stirring uniformly, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 9

Placing 80 parts by weight of orbifloxacin into 2000 parts of ethanol, heating and stirring until the orbifloxacin is dissolved, dissolving 20 parts by weight of tannin into 400 parts of water, combining the two parts of solution, stirring uniformly, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 10

The marbofloxacin hydrochloride and the tannin are crushed, sieved and mixed according to the mass ratio of 95:5, and then are granulated by a wet method and dried to prepare the antibacterial product for animals by a conventional method.

Example 11

Putting 50 parts by weight of marbofloxacin into 200 parts of water, adding hydrochloric acid until the marbofloxacin is completely dissolved, dissolving 50 parts by weight of tannin into 500 parts of water, combining the two parts of solution, uniformly stirring, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 12

Putting 10 parts by weight of ciprofloxacin into 400 parts of water, dissolving 15 parts by weight of tannin into 200 parts of water, combining the ciprofloxacin and the tannin, uniformly stirring, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 13

10 parts of norfloxacin nicotinate is placed in 400 parts of trichloromethane, 15 parts of tannin is dissolved in 200 parts of water, the mixture is slowly added into the solution, stirred, kept at 50 ℃ for reaction for 1 hour, filtered, and the filter cake is dried to obtain the antibacterial product for animals.

Example 14

Dissolving 80 parts by weight of danofloxacin mesylate in 800 parts by weight of water, dissolving 20 parts by weight of tannin in 200 parts by weight of water, combining the two parts of solution, stirring uniformly, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 15

Putting 25 parts by weight of dinofloxacin into 1000 parts of water, dissolving 75 parts by weight of tannin into 750 parts of water, slowly adding the tannin solution into the dinofloxacin, stirring for 10 minutes, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 16

85 parts by weight of lomefloxacin is placed in 1500 parts by weight of water, and aspartic acid is added and stirred until completely dissolved. Dissolving 15 parts by weight of tannin in 150 parts by weight of water, slowly adding the tannin solution into the lomefloxacin solution, stirring for 60 minutes, filtering, and drying a filter cake to obtain the antibacterial product for animals.

Example 17

The enrofloxacin and the tannin are uniformly mixed according to the mass part of 50:50, and the antibacterial product for animals is prepared.

Example 18

Uniformly mixing the ebafloxacin and the tannin according to the mass ratio of 75:25 to prepare the antibacterial product for animals.

Example 19

Healthy pregnant sows 90 (10 weeks gestation), were randomly and evenly divided into 3 groups, and the administration mode: the medicine is administered by drinking water. Setting a blank control group: the drinking water is clear water without any substances; enrofloxacin group: adding enrofloxacin into drinking water, wherein the concentration of enrofloxacin is 0.1g/L (100 ppm); example 1 group of the present invention: the antibacterial product for animals prepared in the invention example 1 is added into drinking water, and the concentration is 0.1g/L (100ppm) according to enrofloxacin.

3 days of pre-experiment (groups without drug addition), 3 groups were recorded for water intake, essentially identical with no difference. On day four, the addition was observed for 12 hours and the water intake was recorded for each group as shown in table 1:

TABLE 1

The experimental results show that: pregnant sows did not receive drinking water containing enrofloxacin at all, while the addition of the product of example 1 to the drinking water had little effect on the water intake of pregnant sows.

Example 20

90 lactating sows (4 weeks postpartum) were randomly and evenly divided into 3 groups, and the administration mode was: the medicine is administered by drinking water. Setting a blank control group: the drinking water is clear water without any substances; danofloxacin mesylate: adding danofloxacin hydrochloride into the drinking water, wherein the concentration of the danofloxacin hydrochloride is 0.05g/L (50 ppm); example 2 group of the present invention: the antibacterial product for animals prepared in example 2 of the present invention was added to drinking water at a concentration of 0.05g/L (50ppm) in terms of danofloxacin.

3 days of pre-experiment (groups without drug addition), 3 groups were recorded for water intake, essentially identical with no difference. On day four, the addition was observed for 12 hours and the water intake was recorded for each group as shown in table 2:

TABLE 2

The experimental results show that: the lactating sows are not fond of drinking water containing danofloxacin mesylate, and the addition of the product of example 2 to the drinking water has little effect on the water intake of the lactating sows.

Example 21

Average weight of 50kg of 36 pigs with bacterial diarrhea, randomly and evenly divided into 3 groups, administration mode: the medicine is administered by drinking water. Setting a blank control group: the drinking water is clear water without any substances; ciprofloxacin hydrochloride group: ciprofloxacin hydrochloride is added into drinking water, and the concentration is 0.05g/L (50ppm) according to ciprofloxacin; example 3 groups of the present invention: the antibacterial product for animals prepared in the invention example 3 was added to drinking water, and the concentration was 0.05g/L (50ppm) in terms of ciprofloxacin.

3 days of pre-experiment (groups without drug addition), 3 groups were recorded for water intake, essentially identical with no difference. On day four, the addition was observed for 12 hours and the water intake was recorded for each group as shown in table 3:

TABLE 3

The experimental results show that: the sick pigs do not like drinking the drinking water containing ciprofloxacin hydrochloride, but the addition of the product in the embodiment 3 to the drinking water hardly influences the drinking water amount of the sick pigs, and can obviously treat the diarrhea of the sick pigs.

Example 22

120 sows at 3 weeks gestation were randomly and evenly divided into 3 groups, and the administration mode was: the medicine is administered by drinking water. Setting a blank control group: the drinking water is clear water without any substances; marbofloxacin group: marbofloxacin is added into drinking water, and the concentration is 0.1g/L (100ppm) according to the marbofloxacin; example 4 groups of the present invention: the antibacterial product for animals prepared in the invention example 4 is added into drinking water, and the concentration is 0.1g/L (100ppm) in terms of marbofloxacin.

3 days of pre-experiment (groups without drug addition), 3 groups were recorded for water intake, essentially identical with no difference. On day four, the addition was observed for 12 hours and the water intake was recorded for each group as shown in table 4:

TABLE 4

The experimental results show that: pigs do not like drinking water containing marbofloxacin, whereas the addition of the product of example 4 to the drinking water has no effect on its water intake.

Example 23

120 healthy backup sows with the average weight of 80kg are randomly and averagely divided into 3 groups, and the administration mode is as follows: mixed feeding administration is carried out. Setting a blank control group: the feed is not added with any medicine; sarafloxacin hydrochloride group: the feed is added with sarafloxacin hydrochloride, and the concentration is 0.1g/kg according to sarafloxacin; example 5 groups of the present invention: the antibacterial product for animals prepared in the embodiment 5 of the invention is added into the feed, and the concentration is 0.1g/kg according to the sarafloxacin.

3 days from pre-experiment (groups without drug addition), the food intake was recorded for 3 groups, essentially identical with no difference. On day four, the feed was started to add the drug and the feed intake was recorded for each group as shown in table 5:

TABLE 5

Injecting: the feed intake is 24 hours data, and the blank feed is restored after 24 hours in view of healthy growth of the pigs.

The experimental results show that: pigs do not like to eat feed containing sarafloxacin hydrochloride, but the addition of the product of example 5 to the feed had little effect on their feed intake.

Example 24

30 healthy English Duroc boars with the average weight of 75kg are randomly and evenly divided into 3 groups, and the administration mode is as follows: the medicine is administered by drinking water. Setting a blank control group: the drinking water is clear water without any substances; difloxacin hydrochloride group: adding difloxacin hydrochloride into the drinking water, wherein the concentration of the difloxacin hydrochloride is 0.1g/L (100 ppm); example 6 group of the present invention: the antibacterial product for animals prepared in the invention example 6 is added into drinking water, and the concentration of the antibacterial product is 0.1g/L (100ppm) in terms of difloxacin.

3 days of pre-experiment (groups without drug addition), 3 groups were recorded for water intake, essentially identical with no difference. On day four, the additions were observed for 12 hours and the water intake was recorded for each group as shown in table 6:

TABLE 6

The experimental results show that: duroc boars do not like drinking water containing difloxacin hydrochloride, but adding the product of example 6 to the drinking water had little effect on the drinking water volume.

Example 25

90 fattening pigs with average weight of 35kg and developing bacterial diarrhea were randomly and evenly divided into 3 groups, and the administration mode was as follows: the medicine is administered by drinking water. Setting a blank control group: the drinking water is clear water without any substances; group of sarafloxacin: adding sarafloxacin into drinking water, wherein the concentration of the sarafloxacin is 0.1g/L (100 ppm); example 7 groups of the present invention: the antibacterial product for animals prepared in the invention example 7 is added into drinking water, and the concentration is 0.1g/L (100ppm) in terms of sarafloxacin.

3 days of pre-experiment (groups without drug addition), 3 groups were recorded for water intake, essentially identical with no difference. On day four, the additions were observed for 12 hours and the water intake was recorded for each group as shown in table 7:

TABLE 7

The experimental results show that: the sick fattening pigs do not like drinking water containing sarafloxacin, but the drinking water amount is hardly influenced by adding the product in the example 7 into the drinking water, and the diarrhea of the sick pigs can be obviously treated.

Example 26

90 pigs with an average weight of 20kg with bacterial diarrhea were randomly and evenly divided into 3 groups, and the administration mode: mixed feeding administration is carried out. Setting a blank control group: the feed is not added with any medicine; enrofloxacin group: enrofloxacin is added into the feed, and the concentration is 0.1g/kg according to the enrofloxacin; example 8 groups of the present invention: the antibacterial product for animals prepared in the embodiment 8 of the invention is added into the feed, and the concentration is 0.1g/kg according to enrofloxacin.

3 days from pre-experiment (groups without drug addition), the food intake was recorded for 3 groups, essentially identical with no difference. On day four, the feed was started to add the drug and the feed intake was recorded for each group as shown in table 8:

TABLE 8

The experimental results show that: the pigs are not fond to drink the feed containing the enrofloxacin, but the feed added with the product of the example 8 has little influence on the feed intake, and can obviously treat diarrhea of sick pigs.

Example 27

24 cows with acute mastitis, clinical symptoms: palpation of the breast was associated with mild fever, pain, and swelling. The milk has floccule or clot, the milk becomes thin, the pH value is alkaline, the somatic cell count and the chloride content are increased, and the milk is randomly and evenly divided into 4 groups, and the administration mode is as follows: the medicine is administered by drinking water. Blank control group: no medicine is added into the drinking water; injection control group: intramuscular injection, namely taking the commercially available marbofloxacin injection, and carrying out intramuscular injection for 1 time every day and continuously using for 5 days according to 2.5mg (calculated by marbofloxacin) per kilogram of body weight; drinking water control group: taking marbofloxacin powder to prepare drinking water containing marbofloxacin with the concentration of 0.025g/L (25ppm) for free drinking; example 10 groups of the present invention: the antibacterial product for animals prepared in the invention example 10 was added to drinking water at a concentration of 0.025g/L (25ppm) in terms of marbofloxacin, and was allowed to drink freely.

The water intake and treatment effect of each group was observed and recorded as shown in table 9:

TABLE 9

The experimental results show that: the product of the example 10 is added into drinking water, the water consumption of the sick cows is equivalent to that of a blank control group and an injection control group, and is far higher than that of a drinking water control group; and the treatment effect is far better than that of a drinking water control group.

Example 28

30 cases of canine yellow dysentery in a certain canine farm, the sick animals excrete white, off-white or yellow-white stink and loose stool, and the colibacillus infection is diagnosed by a laboratory. Randomized, averaged into 3 groups, dosing regimen: the medicine is administered by drinking water. Setting a blank control group: the drinking water is clear water without any substances; norfloxacin nicotinate group: norfloxacin nicotinate is added into drinking water, and the concentration is 0.05g/L (50ppm) according to norfloxacin; example 13 groups of the present invention: the antibacterial product for animals prepared in the invention example 13 was added to drinking water, and the concentration was 0.05g/L (50ppm) in terms of norfloxacin.

3 days of pre-experiment (groups without drug addition), 3 groups were recorded for water intake, essentially identical with no difference. On day four, the additions were observed for 12 hours and the water intake was recorded for each group as shown in table 10:

watch 10

The experimental results show that: the addition of the product of example 13 to drinking water did not reduce the water intake of olfactory sensitive sick dogs and had a significant therapeutic effect on their diarrhea.

Example 29

Thermogravimetric analysis of the product of the invention and a commercial control

1. Apparatus and method

The instrument comprises the following steps: NETZSCH TG 209F1

The method comprises the following steps: 500-N2-600-air, 27 ℃/10.0(K/min)/500 DEG C

2. Results

(1) Thermogravimetric analysis of tannins (fig. 1):

(2) thermogravimetric analysis of ciprofloxacin (FIG. 2)

(3) Thermogravimetric analysis of antibacterial product of example 12 (FIG. 3)

By comprehensively comparing three thermogravimetric analysis graphs of tannin, ciprofloxacin and the antibacterial product of example 12, the thermogravimetric analysis graph of the product of example 12 is obviously changed, the DTG curve between 225 ℃ and 275 ℃ is smoother, and especially the DTG peak shapes of the ciprofloxacin are completely different at 333 ℃ and around 415 ℃. It is demonstrated that there is a certain force between the tannin and ciprofloxacin in the antibacterial product of example 12, and the force is not simply superimposed.

Example 30

In vitro antimicrobial Activity assay for antimicrobial products

Taking the antibacterial product of example 12 as an example, the Minimum Inhibitory Concentration (MIC) of the product against escherichia coli 1.2385 and staphylococcus aureus 1.2465 was measured and used to evaluate the antibacterial activity of the antibacterial product of the present invention.

1. Experimental methods

(1) Reagent preparation

Example 12 preparation of sample solution: 0.0010g of the powder of example 12 sample was weighed out and dissolved in 1mL of DMSO solution, and shaken by hand until the example 12 sample was completely dissolved in DMSO.

Preparing a ciprofloxacin sample solution: 0.0010g of powder of the ciprofloxacin sample is weighed and dissolved in 1mL of DMSO solution, and the solution is shaken manually until the ciprofloxacin sample is completely dissolved in the DMSO solution.

Preparation of 1M NaOH solution: 40.01g of NaOH powder was weighed out and made up to 1L with distilled water.

Preparation of nutrient broth liquid culture medium: weighing 10.0g of peptone, 3.0g of beef extract and 5.0g of NaCl, diluting to 1L with distilled water, adjusting pH to 7.0 with NaOH, and sterilizing at 120 deg.C under high pressure for 20 min.

Preparation of nutrient broth agar culture medium: weighing 10.0g of peptone, 3.0g of beef extract, 5.0g of NaCl and 15.0g of agar powder, diluting to 1L with distilled water, adjusting pH to 7.0 with NaOH, and sterilizing at 120 deg.C under high temperature and high pressure for 20 min.

(2) The Minimum Inhibitory Concentrations (MIC) of the sample of example 12 and the ciprofloxacin sample against E.coli and S.aureus, respectively, were determined by the two-fold dilution method.

Selecting the lowest working concentration of a sample with the relative survival rate of bacteria lower than 10% in a preliminary experiment as a minimum inhibition concentration area, setting six working concentrations by a factor of 2, preparing a nutrient broth agar culture medium plate with corresponding working concentrations, and planting the nutrient broth agar culture medium plate with 5 multiplied by 10⁵The bacteria were plated on a plate and incubated at 37 ℃ for 18h, and then the bacteriostatic effect was judged by colony counting (FIG. 4).

2. Results of the experiment

The results are shown in FIG. 4 and Table 11:

the minimum inhibitory concentration of the sample of example 12 to Escherichia coli is 0.0032 mug/mL, and when the working concentration of the sample of example 12 is 0.0032 mug/mL, no colony grows in 3 experiments; when the minimum inhibitory concentration of the ciprofloxacin sample to escherichia coli is 0.0064 mug/mL, and the working concentration of the ciprofloxacin sample is 0.0064 mug/mL, no bacterial colony grows in 3 experiments.

In example 12, when the minimum inhibitory concentration of the sample on staphylococcus aureus is 0.16 mug/mL and the working concentration of the ciprofloxacin sample is 0.16 mug/mL, no colony grows in 3 experiments; when the minimum inhibitory concentration of the ciprofloxacin sample to staphylococcus aureus is 0.32 mug/mL, and the working concentration of the ciprofloxacin sample is 0.32 mug/mL, no colony grows in 3 experiments.

Wherein the staphylococcus aureus and the escherichia coli which are not added with the medicine group grow vigorously and can not be counted; no colonies were observed in the group with the addition of pure medium.

TABLE 11 EXAMPLE 12 minimum inhibitory concentrations of samples and ciprofloxacin samples against E.coli and S.aureus

The experimental results show that: compared with the single use of the ciprofloxacin, the product of the example 12 formed by combining 40 percent (by weight) of ciprofloxacin and 60 percent (by weight) of tannin can reduce the minimum inhibitory concentration to escherichia coli and staphylococcus aureus, thereby reducing the use dosage of the medicament under the condition of not reducing the medicament effect and reducing the risk of side effects or adverse reactions.

Example 31

In vitro antibacterial activity assay of quinolone and tannin combined antibacterial product

The same procedures as in example 6 were repeated except for replacing the difloxacin hydrochloride in example 6 with ciprofloxacin, wherein the mass ratio of ciprofloxacin to tannin was 20/80, to obtain a product 31A; replacing the difloxacin hydrochloride in the example 6 with levofloxacin, wherein the mass ratio of ciprofloxacin to tannin is 20/80, and carrying out the rest operations in the same way as the example 6 to obtain a product 31B; the same procedures as in example 6 were repeated except for using sarafloxacin to replace the difloxacin hydrochloride in example 6, wherein the mass ratio of ciprofloxacin to tannin is 20/80, thereby obtaining a product 31C; replacing the difloxacin hydrochloride in the example 6 with orbifloxacin, wherein the mass ratio of ciprofloxacin to tannin is 20/80, and performing the rest operations in the same way as the example 6 to obtain a product 31D; the in vitro antibacterial activity of the 4 products was determined

The determination of the Minimum Inhibitory Concentration (MIC) of the product on Escherichia coli is used for evaluating the antibacterial activity of the product. The Minimum Inhibitory Concentration (MIC) was measured by the double dilution method, and a Fractional Inhibitory Concentration (FIC) index was calculated.

Preliminarily determining the minimum inhibitory concentration region of the product to Escherichia coli (figure 5), selecting the minimum working concentration of the product with relative survival rate of bacteria lower than 10% in the preliminary experiment as the minimum inhibitory concentration region, and advancing by a factor of 2Setting three working concentrations, designing two working concentrations backwards, preparing nutrient broth agar culture medium plate with corresponding working concentrations, and planting 5 × 10⁵The bacteria are cultured on a flat plate at a constant temperature of 37 ℃ for 18h, and then the bacteriostatic effect is judged by a colony counting method.

The results are shown in Table 12: when the minimum inhibitory concentration of ciprofloxacin to escherichia coli is 0.0064 mug/mL and the working concentration of ciprofloxacin is 0.0064 mug/mL, no bacterial colony grows in 3 experiments; when the minimum inhibitory concentration of the product 31A to escherichia coli is 0.008 mug/mL and the working concentration of the product 31A is 0.008 mug/mL, no colony grows in 3 tests; when the minimum inhibitory concentration of the levofloxacin on escherichia coli is 0.032 mug/mL and the working concentration of the levofloxacin is 0.032 mug/mL, no bacterial colony grows in 3 experiments; when the minimum inhibitory concentration of the product 31B to escherichia coli is 0.04 mug/mL and the working concentration of the product 31B is 0.04 mug/mL, no colony grows in 3 experiments; when the minimum inhibitory concentration of the sarafloxacin to the escherichia coli is 0.032 mug/mL and the working concentration of the levofloxacin is 0.032 mug/mL, no bacterial colony grows in 3 experiments; when the minimum inhibitory concentration of the product 31C to escherichia coli is 0.08 mug/mL and the working concentration of the product 31C is 0.08 mug/mL, no colony grows in 3 experiments; when the minimum inhibitory concentration of orbifloxacin to escherichia coli is 0.08 mug/mL and the working concentration of orbifloxacin is 0.08 mug/mL, no colony grows in 3 experiments; when the minimum inhibitory concentration of the product 31D to escherichia coli is 0.2 mug/mL and the working concentration of the product 31D is 0.2 mug/mL, no colony grows in 3 experiments; the Escherichia coli can still grow when the working concentration of the tannin sample is 1000 mug/mL, and colonies grow in 3 experiments, so that the lowest bacteriostatic concentration of the tannin sample can exceed 15.6 ten thousand times of that of the ciprofloxacin sample. Wherein the flat escherichia coli without the medicine group grows vigorously and cannot be counted; no colonies were observed in the group with the addition of pure medium.

TABLE 12 minimum inhibitory concentration and FIC of samples on E.coli

The experimental results show that:

the minimum inhibitory concentration of product 31A was 0.008 μ g/mL, MIC according to FIC_{A combined action}/MIC_{A acts alone}+MIC_{Combined action of B}/MIC_{B acts alone}The calculation result shows that the two components in the product 31A show synergistic effect.

The minimum inhibitory concentration of the product 31B is 0.04 mug/mL. According to FIC ═ MIC_{A combined action}/MIC_{A acts alone}+MIC_{Combined action of B}/MIC_{B acts alone}The calculation result shows that the two components in the product 31B show synergistic effect.

The minimum inhibitory concentration of the product 31C is 0.08 mu g/mL. According to FIC ═ MIC_{A combined action}/MIC_{A acts alone}+MIC_{Combined action of B}/MIC_{B acts alone}The calculation result shows that the two components in the product 31B show synergistic effect.

The minimum inhibitory concentration of product 31D was 0.2. mu.g/mL. According to FIC ═ MIC_{A combined action}/MIC_{A acts alone}+MIC_{Combined action of B}/MIC_{B acts alone}The calculation result shows that the two components in the product 31B show synergistic effect.

The products 31A, 31B, 31C and 31D have synergistic effect, and can reduce the using dosage of the quinolone medicaments and maintain the close antibacterial action.

Example 32

The operation was performed in the same manner as in example 7 except that neruoxacin was used in place of sarafloxacin in example 7, wherein the mass ratio of neruoxacin to tannin was 20/80, to obtain product 32, and the in vitro antibacterial activity of the product was measured.

The determination of the Minimum Inhibitory Concentration (MIC) of product 32 against staphylococcus aureus was used to evaluate the antibacterial activity of the product. The Minimum Inhibitory Concentration (MIC) was measured by the double dilution method, and a Fractional Inhibitory Concentration (FIC) index was calculated.

Preliminary determination of the product against Staphylococcus aureusSelecting the lowest working concentration of the product with the relative survival rate of bacteria lower than 10% in the preliminary experiment as the lowest bacteriostatic concentration area, setting three working concentrations forwards by multiple of 2, designing two working concentrations backwards, preparing nutrient broth agar culture medium plates with corresponding working concentrations, and planting the nutrient broth agar culture medium plates with 5 multiplied by 10⁵The bacteria are cultured on a flat plate at a constant temperature of 37 ℃ for 18h, and then the bacteriostatic effect is judged by a colony counting method.

The results are shown in Table 13: the minimum inhibitory concentration of the nervoxacin to staphylococcus aureus is 0.05 mu g/mL, and the minimum inhibitory concentration of the product 16 to staphylococcus aureus is 0.05 mu g/mL; the minimum inhibitory concentration of the tannin sample to staphylococcus aureus is 250 mug/mL. Wherein the flat escherichia coli without the medicine group grows vigorously and cannot be counted; no colonies were observed in the group with the addition of pure medium.

TABLE 13 minimum inhibitory concentration and FIC for samples against Staphylococcus aureus

The experimental results show that:

the minimum inhibitory concentration of the nemonoxacin sample to staphylococcus aureus is 0.05 mu g/mL, the minimum inhibitory concentration of the tannin sample to staphylococcus aureus is 250 mu g/mL, and the inhibitory action is extremely low.

The minimum inhibitory concentration of product 32 was 0.1 μ g/mL. According to FIC ═ MIC_{A combined action}/MIC_{A acts alone}+MIC_{Combined action of B}/MIC_{B acts alone}The calculations show that the two ingredients in product 32 exhibit a synergistic effect.

The product 32 has a remarkable synergistic effect, and can reduce the using dosage of quinolone medicaments and maintain a close antibacterial effect.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. Use of a composition for the manufacture of a medicament for improving the odor or taste, for oral administration to an animal, against bacterial infections, wherein said composition comprises the following components:

(i) a quinolone drug or a pharmaceutically acceptable salt thereof;

(ii) tannin or a pharmaceutically acceptable salt thereof;

the quinolone drugs are selected from enrofloxacin, danofloxacin, marbofloxacin, sarafloxacin, difloxacin, orbifloxacin, dinofloxacin, lomefloxacin or ebafloxacin;

the animal is pig, cattle, sheep, horse, dog or cat;

the improved odor or taste is a reduction or elimination of odor and/or bitterness of a quinolone drug;

the weight ratio of the components (i) and (ii) is 5:95 to 95: 5.

2. The use according to claim 1, wherein said quinolone drug is enrofloxacin.

3. The use according to claim 1, wherein said quinolone drug is danofloxacin.

4. The use according to claim 1, wherein the quinolone drug is sarafloxacin.

5. The use according to claim 1, characterized in that the quinolone drug is orbifloxacin.

6. Use according to claim 1, wherein the weight ratio of components (i) and (ii) is from 15:85 to 85: 15.

7. Use according to claim 1, wherein the weight ratio of components (i) and (ii) is from 25:75 to 75: 25.