CN108976151A - Lauroyl arginine ethyl ester derivative and purposes as animal antibacterial agent - Google Patents

Abstract

The present invention relates to a kind of novel veterinary antibacterial agents, lauroyl arginine ethyl ester ion-pair compound derivative is prepared more particularly to by lauroyl arginine ethyl ester LAE and organic acid reaction, the derivative is as poultry aquatic livestock antibacterial agent, it can prevent and treat the duckling Riemerella anatipestifer disease as caused by riemerella anatipestifer, the peritonitis disease as caused by Escherichia coli and staphylococcus aureus can be treated, the fish septicemia caused by Aeromonas sobria can be prevented and treated, and the antibacterial agent for animals relative to former LAE, it prevents and treats effect and is significantly improved.Prepared derivative in human body and animal body can safe disposal, toxic side effect is small, avoid the generation of the environment drug-fast bacteria caused by being discharged into environment due to remaining antibacterial agent, while reaching effective antibacterial effect to environment negatively affect it is small.

Description

Lauroyl arginine ethyl ester derivative and purposes as animal antibacterial agent

The present patent application requires Chinese invention patent application 201711071006.8, entitled " lauroyl arginine The priority application of the purposes of ethyl ester and its derivative as antibacterial agent for animals ".

Technical field

The present invention relates to a kind of ion-pair compound derivative of lauroyl arginine ethyl ester and preparation method and this spread out Biology is in the application for treating and preventing animal pathogen disease.The especially purposes of derivative preparation animal antibacterial agent.

Background technique

Riemerella anatipestifer disease is caused by riemerella anatipestifer (Riemerellaanatipestifer, RA) A kind of contagious disease, also known as duck infectious serositis, duck septicemia, pest of duck syndrome, Pasteurella anatipestipestifer disease etc..It is more The duckling of 1-8 week old is seen, wherein the duckling of 2-4 week old is most susceptible.In acute or chronic septicemia, it is mainly shown as in clinical aspect The secretion of eye and nose increases, breathes, coughing, diarrhea, incoordination and neck tremble, and neck skew occur in a small number of chronic cases Etc. symptoms.There is arthritis with fibrinous pericarditis, perihepatitis, air bag inflammation, meningitis and some cases in lesion for spy Sign, often causes the large quantities of morbidities and death of duckling.The disease incidence of the disease is up to 90% or more, the death rate and morbidity duck age in days, bacterium Strain virulence, distress factor are related, reach as high as 75%.In addition to 1~8 week old duck can be caused dead, it can also cause fallopian tubal Inflammation, causes adult duck laying rate decline, and growth retardation causes great economic loss to raiser.It is reported for the first time from nineteen eighty-two Since road, this disease has become a kind of most common bacterial disease for endangering meat duck aquaculture.Under field conditions (factors), this disease is throughout the year all There is generation.The wounds through respiratory tract, alimentary canal or skin such as main feed, drinking-water, dust, the droplet for passing through pollution are (especially Sufficient web portion skin) enter in poultry body and causes disease.The duck of different cultivars for example Beijing duck, cherry valley duck, the high duck of Di, teal, Kind duck, Mule duck, sheldrake etc. can infection morbidities.In July, 2012 takes out 48 duckeries in Jiangsu Province to August at random It looks into, wherein there are the appearance of doubtful Riemerella anatipestifer disease case in 45 duckeries, or even has 12 duckeries during selective examination Unexpectedly the disease is broken out twice.In addition, surrounding duck can also break out the disease in succession, thus if the disease occurs for a certain duckery It can be seen that the seriousness that the disease causes damages to duck culturing industry.

Staphylococcus aureus disease refers to the common name for causing many animals various disease or sick type by staphylococcus aureus. Pathogenic staphylococcus can cause various domestic birds and animals to be fallen ill, and young age livestock and poultry are most susceptible to this disease, mostly through alimentary canal sense Dye, chicken also can have diarrhea, enteritis, hepatic necrosis etc. by respiratory tract infection, common sympton.Often cause two class diseases, Yi Leishi Purulent disease mainly causes mammitis, arthritis, wound infection and septicemia of animal etc.；Another kind of is toxin disease, It can be caused Poisoning enteritis and the Toxin shock syndrom of people etc. of animal by the feed that pathogenic bacteria pollute.Staphylococcus aureus Pathogenicity power depend primarily on its generation virulence virulence factor, mainly include plasma-coagulase, enterotoxin, heat-resisting nucleic acid Enzyme, hemotoxin and leukocidin etc..

Colibacillosis refers to the common name for causing many animals various disease or sick type by enteropathogenic E. Coli.Pathogenicity The non-pathogenic Escherichia coli normally to live away from home in Escherichia coli and people and animals' enteron aisle are in form, staining reaction, cultural character and biochemistry Reaction etc. is not different, but antigen construct is different.Enteropathogenic E. Coli can cause various domestic birds and animals to be fallen ill, such as pigs and cattle Sheep eared pheasant rabbit etc., young age livestock and poultry most susceptible disease, mostly infects through alimentary canal, chicken can also pass through respiratory tract infection.Wherein Radix Polygalae Crotalarioidis Bacillosis examines performance also difference in facing for piglet, can be divided into yellow dysentery according to the difference in the growth period of piglet and pathogen serotype Type, dysentery characterized by white mucous stool type and edema type.When yellow dysentery occurs for piglet, 90% or more of ordinary wave and a brood of piglet, case fatality rate is high, and what is had is reachable 100%；The disease incidence of dysentery characterized by white mucous stool is 30%~80%；Oedema disease incidence is 10%~35%.The colibacillosis of chick is usual Show as acute septic, yolk peritonitis, ophthalmia, air bag inflammation Swollen head syndrome etc..Disease incidence is sick up to 30%~60% Dead rate is up to 100%.

Aeromonas sobria is Gram-negative, amphimicrobion, is normally present in various water environments, soil environment In, and be a kind of infecting both domestic animals and human pathogenic bacteria.Aeromonas sobria can generate a variety of virulence factors such as bacteriolysin, ectoenzyme, can Cause aquatic livestock to suffer from septicemia, to cause the death of aquatic livestock, seriously affects the economic base of aquaculture.In addition, Pathogenic bacteria are also possible to be infected by aquatic products to people, and it is food poisoning or sepsis that patient, which will appear the symptoms such as diarrhea even to develop, Disease.

In addition to improving rearing conditions, applying antibiotic is the major measure for preventing and treating beasts, birds and aquatic products bacteriosis. However, in recent years, such as aureomycin, terramycin, tetracycline, chloramphenicol common antibiotics exist because of its disease-resistant, somatotrophic effect It is used without restraint in livestock and poultry breeding industry.According to statistics, the annual antibiotic feedstock capacity in China is about 210,000 tons, wherein having 9.7 ten thousand The antibiotic of ton is used for livestock and poultry breeding industry, accounts for the 46.1% of total growth.Inappropriate antibiotic usage, drug quality are without guarantor Barrier, incomplete supervision and not rigorous medication regulation result in abuse of antibiotics aggravation, to not produce many mistakes still not And serious problems, such as bacterial drug resistance generation, animal body immunity function decline, meat products medicament residue, directly Endanger the health of the mankind.There is research to claim, about 75% antibiotic is can not partially to be remained by human body or animal body Absorption And Metabolism In vivo, antibiotic residue is able to detect that in the live chickens or chicken in jelly tissue for having 20%-50%；Remaining antibiotic also can be with Excreta enter in environment, some meetings are directly entered river and influence the safe drinking water of downstream resident, China's many cities along the river The poultry industries antibiosis such as terramycin, tetracycline, fortimicin, Amoxicillin, aureomycin was all detected in resident's tap water in city The residual of element.There is research to carry out antibiotic detection to 1000, Shanghai children's urine in recent years, is detected in 58% urine sample To a variety of veterinary antibiotics (such as tylosin, duomycin and Enrofloxacin) only used in aquaculture.It is prior It is that these remaining antibiotic can carry out natural selection to the microorganism in environment or induce its gene mutation, has drug resistance The bacteria living of property gets off and continues to breed more drug-fast bacterias.In addition, bacterium can also pass through engagement, conversion, transduction, swivel base Etc. modes the antibiotic resistance gene transfer of oneself is not of the same race to other, belong to microorganism be allowed to obtain drug resistance.This allows for drug resistance Bacterium is enriched in the environment, and contaminated soil, water source easily cause disease if being contacted by people or livestock and accelerate drug resistance The diffusion of bacterium.Other than violation addition antibiotic or antibacterial agent, veterinary drug and feed addictive mainly include preservative, dust-proof Agent, antioxidant, antiprotozoal agent etc..These substances will be brought because of long-time service or improper use to livestock and poultry and health of human body Harm.

Therefore, in aquatic products, herding, the cultivation of poultry, especially in birds (such as chicken and duck) cultivation, there is an urgent need to one kind Can quick sterilization, and degradable, noresidue antibacterial agent in vivo both can effectively prevent disease in aquaculture and occurred and climing Prolong, moreover it is possible to avoid similar influence of the antibiotic residue to human body and environment.

Lauroyl arginine ethyl ester (Ethyl lauroylarginate, LAE) is one kind by fatty acid and binary amino acid Organic matter made of condensation is white hygroscopic solid, and chemical property is stablized in the range of pH3~7,50~58 DEG C of fusing point, should At a temperature of the LAE of 247g be dispersed in the water of 1kg, its distribution coefficient in water and in oil is greater than 10, that is, is primarily present in water Xiang Zhong.The study found that lauroyl arginine ethyl ester LAE have strong antibacterial, bio-toxicity is low, internal metabolic effects are good, with The high feature of Environmental compatibility.And wherein most representative feature is the metabolism noresidue of lauroyl arginine ethyl ester, correlation is ground Natural metabolism can quickly be carried out in human body and animal body by studying carefully display lauroyl arginine ethyl ester, generate lauric acid and arginine, Further it is metabolized as ornithine, urea, carbon dioxide and water.It is generated all in lauroyl arginine ethyl ester metabolic process Metabolite and final product be all it is nontoxic, with the metabolite phase of the food of humans and animals daily ingestion in vivo Together.

For example, Chinese patent application CN201710056593, entitled " a kind of fruit and vegetable preserving antistaling agent and its preparation Methods and applications " are disclosed using lauroyl arginine ethyl ester hydrochloride and soluble metyl hydroxybenzoate as the composition of main active As fruit and vegetable preserving antistaling agent, it can effectively inhibit the bacterial growth for causing fruits and vegetables rotten.However, the invention middle and high concentration The independent fungistatic effect of soluble metyl hydroxybenzoate (2000 μ g/ml) is better than the LAE (1000 μ g/ml) of low concentration, this is because it has There is phenolic hydroxyl structure, anti-microbial property is much better than benzoic acid, sorbic acid, therefore under the premise of guaranteeing antiseptic property, this method It explicitly points out and replaces LAE using soluble metyl hydroxybenzoate, facilitate the dosage cost for reducing preservative.

Chinese patent application CN201510748675, it is entitled " using lauroyl arginine ethyl ester inhibit alcohol hair The method of ferment contaminating microorganisms " discloses the method for inhibiting alcoholic fermentation contaminating microorganisms using lauroyl arginine ethyl ester, should Method includes that LAE and its salt compounds are added in the fermentation liquid of saccharomyces cerevisiae with the concentration lower than 50 μ g/ml, can be effective The growth of lactic acid bacteria inhibiting, and control the growth of other contaminating microorganisms.However, bacteriostatic agent minimal effect to a certain extent The growth of saccharomycete, and alcohol output is caused to reduce by 0.6%.

Chinese patent application CN201610466729, entitled " a kind of mild baby child's hair washing shower bubble " are open The characteristics of a kind of mild baby child has one's hair wash shower bubble, is directed to baby child's hair and skin quality, select cocounut oil acyl disodium glutamate, Cocoamidopropyl betaine and sulfonic acid hydroxypropyl acrylate lauryl glucoside cross-linked polymer sodium compounding are used as surfactant system, Select camellia seed oil, alpha-glucans oligosaccharides/inulin compound as conditioning ingredients, Flos Chrysanthemi Indici extract and lauroyl arginine second Ester HCl compounding is used as protective system, and each raw material cooperates with each other in the invention, and cleaning effect is good, mild non-stimulated.

Chinese patent application CN201280073013, entitled " antimicrobial of synergistic effect " be open pass through by A effective amount of N- α-long alkanoyl binary amino acid alkyl ester salt is combined with fatty acid monoglyceride provides the anti-of synergistic effect Microbial composite generates more effective antimicrobial and food preservative.Meanwhile Chinese patent application The composition of CN200810131638, entitled " microbicide composition " open methylisothiazolinone and LAE are used In the purposes for preparing antimicrobial and food preservative.However, this method is related to a variety of antipathogenic compositions including LAE, The independent bacteriostasis of LAE is not studied.Meanwhile the invention only only teach the composition for daily product, detergent, The purposes such as wound care compositions, varieties of food items, all kinds of medical cleaning products do not teach how to use single LAE ingredient In the purposes of the antibiotic property feed of poultry aquatic products.

Chinese patent application CN201280027864, entitled " the makeup sun-screening agent with improved water resistance Or dermatology's sun-screening agent " purposes that a kind of LAE is used to prepare makeup sun-screening agent or dermatology's sun-screening agent, said preparation are disclosed It further include -10 stearate of emulsifier polyglycerol other than UV lightscreening agent.

As the immediate prior art, Chinese patent application CN200580051259, entitled " including cation The first public LAE of the protective system of surfactant " and its hydrochloride are for the purposes in protective system, in food, cosmetics The middle system of addition comprising 0.2g/kg LAE is to play antisepsis.The invention research antifungal mechanism of LAE, and mention How LAE is used for the application of the antisepsises such as food, cosmetics out, therefore food safety office of the U.S. to be in the approval moon in 2005 Osmanthus acyl arginine ethyl ester is used for food preservative；Food safety office of European Union, Australia and New Zealand also all have approved within 2012 Lauroyl arginine ethyl ester is used for food preservative.Meanwhile it being put forward for the first time the application in terms of cosmetics in view of the invention, it is subsequent In research find lauroyl arginine ethyl ester can be used for oral care in terms of product (such as US20100330136A1, EP2361606A2, EP231603A2), the formation of dental plaque in oral cavity can be effectively suppressed in such as mouthwash, toothpaste, it with gargle Other chemical components in water are compatible and chemical property is stablized；The change that lauroyl arginine ethyl ester can be used for having local treatment effect In cosmetic, these cosmetics have characteristic below: antibacterial effect, and low toxicity without sensitization, does not stimulate skin.Currently, Researcher is developing clean hand cleanser and the bacteriostatic agent for skin surface.

Summary of the invention

In conclusion existing invention is not taught how using single lauroyl arginine ethyl ester (LAE) derivative The purposes of antibiotic property drug as poultry aquatic products, also not publicly lauroyl arginine ethyl ester (LAE) derivative as antibacterial The suitable concentration of property drug.Therefore, the present invention utilizes lauroyl arginine ethyl ester and its derivative or its hydrate (preferably LAE) As the fungistatic effect of preservative, on the basis of first patent application of the LAE of the applicant as antibacterial agent for animals, develop New LAE derivative is no longer limited to selection and is traditionally suitable for LAE wherein breaking through the conventional thought for derivative exploitation Acid, alkali, salt/ester convenient form, or to LEA carry out acid, alkali, salt or esterified group processing, but creativeness selection A kind of sulfonate groups of the antibacterial synergistic effect that LAE can be enhanced, the energy matter that can significantly supplement the nutrients again simultaneously, and by the two New derivative, i.e. ion-pair compound are combined by intermolecular strong ionic bond unconventionally, to improve significantly LAE derivative is in the purposes as antibacterial agent for animals.

First purpose of the invention is to provide the lauroyl arginine ethyl ester (LAE) with the structural formula as shown in following formula (I) Derivative:

Wherein,

X is the organic acid RCOO with bacteriostasis, energy effect^-；It is selected from salicylic acid, formic acid, acetic acid, diacetic acid, Propionic acid, propionic acid, butyric acid, lactic acid, benzoic acid, sorbic acid, fumaric acid, citric acid, tartaric acid, malic acid, phosphoric acid, oxalic acid or carbonic acid It is arbitrary one or more of；

R₁It is that the linear saturated fatty acids group containing 8-14 carbon atom or the straight chain containing 8-14 carbon atom are oxygen-containing Acid groups.

R₂It is the straight chain fatty acid groups containing 1-18 carbon atom or the Branched fatty acidic group containing 1-18 carbon atom Group or the aromatic group containing 1-18 carbon atom, or the straight chained alkyl containing 1-4 carbon atom.

R₃It is one kind of having structure.

The range of n is 0-4.

In one embodiment, the RCOO^-Selected from salicylic acid, formic acid, acetic acid, diacetic acid, propionic acid, propionic acid, butyric acid, Lactic acid, benzoic acid, sorbic acid, fumaric acid, citric acid, tartaric acid, malic acid, phosphoric acid, oxalic acid or carbonic acid acid group.

In another embodiment, organic acid RCOO^-Selected from niacin, tartaric acid, oxalic acid acid group.

In a specific embodiment, the present invention provides the lauroyl essence ammonia with the structural formula as shown in following formula (III) Acetoacetic ester (LAE) organic acid ion is to compound:

RCOO^-For the acid group of acid as described below: being selected from salicylic acid, formic acid, acetic acid, diacetic acid, propionic acid, propionic acid, butyric acid, cream Acid, benzoic acid, sorbic acid, fumaric acid, citric acid, tartaric acid, malic acid, phosphoric acid, oxalic acid or carbonic acid.

In another embodiment, organic acid RCOO^-Selected from niacin, tartaric acid, oxalic acid acid group.

Second purpose of the invention is to provide above-mentioned LAE derivative and is used to prepare the cause for treating or preventing poultry, aquatic livestock The purposes of the drug of germ infection.

In one embodiment, the drug is for treating or preventing duckling Infective Gram-negative bacteria Drug, and the drug is administered with the dosage that every kg body weight gives 4-40mg formula (I) or (III) compound represented. In a preferred embodiment, which gives the agent of 8-16mg formula (I) or (III) compound represented with every kg body weight Amount is administered, and the Pathogenic gram-negative bacterium is riemerella anatipestifer.In the most preferred embodiment, the drug with Every kg body weight gives 16mg formula (I) or the dosage of (III) compound represented is administered to treat in duckling infection pest of duck The disease of Mo Shi bacillus, or give the dosage of 8mg formula (I) or (III) compound represented and be administered to prevent duckling infection The disease of riemerella anatipestifer.

In another embodiment, the drug is the pathogenic Gram for treating or preventing poultry, aquatic livestock The drug of negative bacterial diseases, and the drug gives 0.625-10mg formula (I) or (III) compound represented with every kg body weight Dosage be administered.In a preferred embodiment, the dosage is 2.5-10mg, the Pathogenic gram-negative bacterium It is inner Mo Shi bacillus, Escherichia coli, Pasteurella, salmonella, haemophilus, brucella or Aeromonas sobria.

In other embodiments, the drug is the gram-positive bacteria for treating or preventing poultry, aquatic livestock The drug of disease, and the drug with every kg body weight give the dosage of 2.5-25mg formula (I) or (III) compound represented into Row application.In preferred embodiments, the gram-positive bacteria is staphylococcus aureus, streptococcus, erysipelas bacillus, branch Bacillus, bacillus anthracis, and the drug gives 2.5,10 or 25mg formula (I) or (III) compound represented with every kg body weight Dosage be administered.

Also in other embodiments, the drug is the drug for treating grice diarrhoea, and the drug is with every thousand Gram weight gives 10-50mg formula (I) or the dosage of (III) compound represented is administered piglet.In preferred embodiments, The drug is administered piglet with the dosage that every kg body weight gives 30-50mg formula (I) or (III) compound represented, and Obtain the effect better than the treatment grice diarrhoea with dosage antibiotic.In the most preferred embodiment, the drug is with every thousand Gram weight gives 50mg formula (I) or the dosage of (III) compound represented is administered piglet, and obtains better than with dosage antibiosis The effect of the treatment grice diarrhoea of element.

In another embodiment, the dosage form of the drug comprising compound represented includes granule, solution, hangs Floating agent, pulvis, capsule, finish, paste.

In any of the above-described embodiment, the poultry, aquatic livestock include: chicken and duck goose turkey quail pigeon, pig, cattle and sheep Horse camel cat and dog and fish shrimp crab etc., the pathogenic bacteria include but is not limited to yellowish-white dysentery, asthma, erysipelas disease, oedema, Clostridium property enteritis, Proliferative Enteritis, tuberculosis, pasteurellosis, bacillus anthracis disease, salmonellosis.

The invention also provides derivatives shown in formula (I) or (III) in preparation for changing the cell membrane pole of microorganism Application in the drug of property.

In a preferred embodiment, the compound represented can kill pathogenic microorganisms in 30min, and The appearance of drug-fast bacteria is not caused.In another preferred embodiment, the compound represented can be stimulated and be caused a disease for continuous 30 days Appearance of the microorganism without causing drug-fast bacteria.In a more preferred embodiment, the compound represented is dynamic to normal lactation Object cytotoxicity is low.Do not cause erythrocyte hemolysis under minimum bactericidal concentration.

In another embodiment, the compound represented on healthy duckling survival rate without influence, to healthy young Duck body weight increase is without influence, to healthy duckling internal organs nontoxicity, and can reduce scorching in the animal body risen due to bacterium infection Inflammation factor IL-1 β and/or IL-1 β protein level.

Third purpose of the present invention is to provide the lauroyl arginine ethyl ester derivative as shown in formula (III) or its hydrate Or pharmaceutically acceptable salt, for changing the polar method of cell membrane of microorganism, step includes:

(1) invasive organism solution is diluted to OD600=0.05, cell membrane polar dies is then added；

(2) derivative solution shown in formula (I) or (III) is added until final concentration of 16 μ g/ml, sufficiently reacts；

(3) using the light of 622nm wavelength as exciting light, the fluorescent value at 670nm is measured；

(4) fluorescent value for passing through each sample of flow cytomery, obtains bacterial cell membrane by ratio calculated The degree of depolarising.

4th purpose of the invention is to provide the system of lauroyl arginine ethyl ester (LAE) derivative as shown in formula (III) Preparation Method, step include:

(1) formula (II) compound represented is dissolved by heating, acylate solution is then added；

(2) mixing is sufficiently stirred, and under heating conditions, passes through lauroyl essence shown in following reaction equations raw formula (III) Propylhomoserin ethyl ester ion-pair compound；

(3) sufficiently after reaction, cooling room temperature is dried in vacuo, after purification to prepare lauroyl essence ammonia shown in formula (II) Acetoacetic ester organic acid ion is to compound.

In one embodiment, organic acid RCOO^-Acid group selected from following acid: salicylic acid, formic acid, acetic acid, double second Sour sodium, propionic acid, butyric acid, lactic acid, benzoic acid, sorbic acid, fumaric acid, citric acid, tartaric acid, malic acid, phosphoric acid, oxalic acid or carbon Acid；And selected from niacin, pantothenic acid, folic acid, the ascorbic acid having for animal extra-nutrition energy.In a preferred embodiment In, the organic acid is selected from the acid group of niacin, tartaric acid, oxalic acid, carbonic acid.

In another embodiment, the organic acid is added in methanol solution, and suitable NaOH is added, be stirred at room temperature Until white solid is precipitated, filter and is washed with methanol point, acylate is obtained.

The beneficial effects of the present invention are:

Research has shown that, niacin belongs to B family vitamin, and niacin participates in HypercholesterolemicRats, the oxidation process of tissue respiration and The process of carbohydrate anaerobic decomposition.Mainly by plant, Microbe synthesis, (certain animals can also realize that itself is closed by tryptophan At, but synthetic quantity is far from enough needed for itself), in animal body can not mass storage but be that growth of animal institute is indispensable Substance.Ruminant can be provided by the microorganism in cud, and poultry and pig then rely on feed to carry out external source supplement.Niacin Deficiency disease has following performance: poultry shows feather rareness and fluffy matt skin and the short and thick leg of foot dermatitis bone are bent；Domestic animal Show dermatitis, loss of appetite and syntexis.According to the catalogue of feed additive varieties of China 2013 editions, niacin belongs to vitamins addition Agent can be used for various cultivated animals, be conducive to the healthy growth of cultivated animals.

Tartaric acid is to be present in a kind of organic acid in various plants and the main organic acid in grape wine, is usually used in Food addition plays antioxidant.Oxalic acid is also a kind of organic acid being present in many green plantss, such as spinach, red In addition potato, taro and rheum officinale etc. also contain a large amount of oxalic acid in cocoa bean.Therefore the present invention wishes that LAE is nontoxic very with these Ion pair is synthesized to the organic acid for being conducive to animal health, to promote the disease-resistant or growth promotion function of LAE.

By experiment, select has certain bacteriostasis, plays nutrition (such as niacin) or energy (oxalic acid) effect the present invention Or as organic acid needed for biological metabolism, single ion-pair compound is generated by condensation reaction and LAE, list can be changed The physicochemical properties such as body molecular melting, stability, can both play respective activity, also can produce synergistic effect, to improve biology Availability further promotes curative effect of medication；It readily penetrates through lipophilic cell membrane again simultaneously, improves and absorb.

Present invention demonstrates that derivative shown in formula (III) relative to LAE compound shown in formula (I), also has as follows Advantage:

1, derivative shown in formula of the invention (III) will not generate inhibition to the antibacterial activity of the former LAE of single component Effect, is beneficial to antibacterial activity on the contrary.Wherein, niacin ion pair derivative produces staphylococcus aureus compared to LAE Significant fungistatic effect.

2, derivative successive administration shown in formula of the invention (III) does not induce the medicament-resistant mutation of pathogenic microorganisms for 30 days, And low relative to commercially available antibiotic induction bacterial resistance ability, the derivative is resistance to relative to LAE induction staphylococcus aureus The reduced capability of medicine.

3, derivative shown in formula of the invention (III) does not cause Haemolysis under minimum bactericidal concentration.

4, derivative shown in formula of the invention (III) is able to ascend the duckling survival for having infected riemerella anatipestifer bacillus Rate, reduces the risk of duckling infection riemerella anatipestifer bacillus, and spreads out on healthy duckling survival rate without influence, and compared to LAE The effect of biology, dosage is lower, and therapeutic effect is more preferable

5, derivative shown in formula of the invention (III) is significantly lower than the production of LAE as the effective dosage ranges of drug The upper dosage limit range 40mg/kg bw needed

6, derivative shown in formula of the invention (III) can restore the inflammation due to caused by bacterium infection in animal body Factor IL-1 β, TNF-α protein level raising.

7, it is demonstrated experimentally that administration is discontinued for 24 hours after 3 months, the heart, liver, spleen, lung, kidney, small intestine, stomach, the muscle of administration group animal Tissue drug residue meets requirement of the European Union to non-regulation Residual Veterinary Medicines.

8, it is demonstrated experimentally that derivative shown in the formula (III) can prevent and treat the bacteriosis of aquatic livestock, additionally it is possible to Promote the nutrient growth of poultry aquatic livestock, in which: anti-infective experimental result is shown, is eaten not containing derivative shown in formula (III) The test fish of object feed is all dead, eat the Tilapia mossambica survival rate containing derivative shown in formula (III) up to 91.67 ± 2.89,88.33 ± 7.63 survival rates higher than LAE；For Tilapia mossambica, can be improved body weight increase rate 3.52%, 6.91%, 4.64% and specific growth rate 7.94%, 13.08%, 6.54%, feed coefficient 4.76%, 5.56%, 2.38% is reduced, it is high In the body weight increase rate 3.03%, 6.23%, 3.88% and specific growth rate 12.32% of LAE, feed coefficient 3.15% is reduced.

9, it is demonstrated experimentally that compared with the gentamicin with dosage, formula (III) compound represented can be controlled more effectively Grice diarrhoea is treated, and avoids the use of antibiotic, reduces the residual of the antibiotic in livestock.

10, lauroyl arginine ethyl ester derivative in human body and animal body can safe disposal, toxic side effect is small, avoids The generation of environment drug-fast bacteria caused by being discharged into environment due to remaining antibacterial agent, while reaching effective antibacterial effect Environment is negatively affected small.

Detailed description of the invention

The ESI mass spectrogram of the cationic B+ molecular ion peak of Fig. 1: LAE ion-pair compound；

The ESI mass spectrogram of the anion A- molecular ion peak of Fig. 2: LAE niacin ion-pair compound；

The peak shape and chemical shift figure of Fig. 3: LAE 1H-NMR；

Fig. 4: the 1H-NMR of niacin peak shape and chemical shift figure；

The peak shape and chemical shift figure of the 1H-NMR of Fig. 5: LAE niacin ion pair；

The ESI mass spectrogram of the anion A- molecular ion peak of Fig. 6: LAE tartaric acid ion-pair compound.

Specific embodiment

In conjunction with following specific embodiments and attached drawing, the present invention is described in further detail, protection content of the invention It is not limited to following embodiment.Without departing from the spirit and scope of the invention, those skilled in the art it is conceivable that change Change and advantage is all included in the present invention, and using appended claims as protection scope.Implement process of the invention, Condition, reagent, experimental method etc. are among the general principles and common general knowledge in the art in addition to what is specifically mentioned below, There are no special restrictions to content by the present invention.

Embodiment one: lauroyl arginine ethyl ester hydrochloride synthesizes the preparation method of ion-pair compound with niacin

Naotin (being purchased from uncommon love (Shanghai) the chemical conversion industry Development Co., Ltd of ladder) 2.0g is dissolved in 50mL water, is configured to Niacin sodium-salt aqueous solution (A)；Lauroyl arginine ethyl ester hydrochloride 6.8g is dissolved in 40mL water, is heated to 90 DEG C, until the moon Osmanthus acyl arginine ethyl ester hydrochloride all dissolves, and lauroyl arginine ethyl ester hydrochloride aqueous solution (B) is made；Under the conditions of 90 DEG C Niacin sodium-salt aqueous solution (A) is slowly added into lauroyl arginine ethyl ester hydrochloride aqueous solution (B), is stirred continuously, reaction 2 Hour, it is cooled to room temperature, filters, precipitating is sufficiently washed with pure water, precipitate 60 DEG C of vacuum drying to get niacin ion pair chemical combination Object 7.6g.

The analysis of embodiment two lauroyl arginine ethyl ester niacin ion-pair compound molecular formula, molecular weight

By mass spectrum,¹H-NMR、¹³The obtained compound molecule formula of C-NMR Spectrum Analysis are as follows:

1. mass spectrum (ESI) is analyzed

Cationic B⁺The m/z=385.3 of molecular ion peak, referring to Fig. 1；

Mass Spectrometer Method ESI+ is 124.2, referring to fig. 2.Then ESI- is 122.2, i.e. anion A^-The m/z=of molecular ion peak 122.2。

Cationic calculated value is 507.4 in niacin ion-pair compound, measured value and theoretical calculation.

2.NMR analysis

By lauroyl arginine ethyl ester hydrochloride (referring to Fig. 3), niacin¹H-NMR (referring to fig. 4) and ion-pair compound 's¹H-NMR (referring to Fig. 5) compares.Since LAE ion-pair compound is in salification process, the moon in the ion-pair compound The peak shape of osmanthus acyl arginine ethyl ester and chemical shift variation are little, but all hydrogen on niacin have change in displacement, and wave spectrum is special Compared with sign is with original inorganic acid salt (i.e. LAE hydrochloride), soda acid two parts space length is more nearly, and is had an impact, thus its with Former LAE and its hydrochloride are compared, and are generated corresponding change, are not the two-part superposition of simple soda acid, such as in pure washing When washing precipitating, dissolubility has changed, this illustrates to produce between all protons of lauroyl arginine ethyl ester and niacin strong Interaction, and stable single compound structure is formd by strong ionic bond.

Embodiment three: lauroyl arginine ethyl ester hydrochloride synthesizes the preparation method of ion-pair compound with tartaric acid

Tartaric acid (being purchased from uncommon love (Shanghai) the chemical conversion industry Development Co., Ltd of ladder) 2.0g is dissolved in 50mL methanol, is added The NaOH of equivalent is stirred at room temperature until white solid is precipitated, and suction filtration is simultaneously washed with 30mL methanol in three times, obtains sodium tartrate Salt.Sodiotartrate is dissolved in 50mL water, is configured to sodium tartrate saline solution (A)；By lauroyl arginine ethyl ester hydrochloride 5.6g is dissolved in 40mL water, is heated to 90 DEG C, until lauroyl arginine ethyl ester hydrochloride all dissolves, lauroyl essence ammonia is made Acid ethyl ester hydrochloride saline solution (B)；Sodium tartrate saline solution (A) is slowly added into lauroyl arginine under the conditions of 90 DEG C It in ethyl ester saline solution (B), is stirred continuously, reacts 2 hours, be cooled to room temperature, filter, it is heavy sufficiently to be washed with pure water It forms sediment, precipitates 60 DEG C of vacuum drying to get tartaric acid ion-pair compound 6.3g.

The analysis of example IV lauroyl arginine ethyl ester tartaric acid ion-pair compound molecular weight

Mass spectrum (ESI) analyzes cation B⁺The m/z=385.3 of molecular ion peak (referring to Fig. 1)

Anion A^-The m/z=149.0 of molecular ion peak (referring to Fig. 6)

Cationic calculated value is 534.3 in niacin ion-pair compound, measured value and theoretical calculation.

Embodiment five: lauroyl arginine ethyl ester hydrochloride synthesizes the preparation method of ion-pair compound with oxalic acid

1.0g (is purchased from and explores Co., Ltd) by oxalic acid to be dissolved in 50mL methanol, the NaOH of equivalent is added, and is stirred at room temperature straight To white solid is precipitated, filter and is washed in three times with 30mL methanol, oxalic acid sodium salt is obtained.Oxalic acid sodium salt is dissolved in 50mL water, It is configured to oxalic acid sodium-salt aqueous solution (A)；Lauroyl arginine ethyl ester hydrochloride 4.7g is dissolved in 40mL water, is heated to 90 DEG C, Until lauroyl arginine ethyl ester hydrochloride all dissolves, lauroyl arginine ethyl ester hydrochloride aqueous solution (B) is made；At 90 DEG C Under the conditions of oxalic acid sodium-salt aqueous solution (A) is slowly added into lauroyl arginine ethyl ester hydrochloride aqueous solution (B), constantly stir Mix, react 2 hours, be cooled to room temperature, filter, precipitating is sufficiently washed with pure water, precipitate 60 DEG C be dried in vacuo to get oxalic acid from Son is to compound 5.0g.

According to the method for embodiment two, NMR analysis and ESI analysis are carried out, the results showed that the wave spectrum of the ion-pair compound Feature is not the two-part superposition of simple soda acid, and soda acid two parts space length is close, is had an impact, spectral data and original LAE and its hydrochloride are compared, and generate corresponding change, such as in pure water washing precipitating, dissolubility has changed, this says Strong interaction is produced between all protons and oxalic acid of bright lauroyl arginine ethyl ester, and is formed by strong ionic bond Stable single compound structure.

Embodiment six: lauroyl arginine ethyl ester hydrochloride synthesizes the preparation method of ion-pair compound with carbonic acid

1.0g (is purchased from and explores Co., Ltd) by sodium carbonate to be dissolved in 50mL water, is configured to aqueous sodium carbonate (A)；By the moon Osmanthus acyl arginine ethyl ester hydrochloride 4.0g is dissolved in 40mL water, is heated to 90 DEG C, until lauroyl arginine ethyl ester hydrochloride is complete Portion's dissolution, is made lauroyl arginine ethyl ester hydrochloride aqueous solution (B)；It is under the conditions of 90 DEG C that aqueous sodium carbonate (A) is slow It is added in lauroyl arginine ethyl ester hydrochloride aqueous solution (B), is stirred continuously, react 2 hours, be cooled to room temperature, filter, use Pure water sufficiently washs precipitating, precipitates 60 DEG C of vacuum drying to get carbonate ions to compound 4.0g.

According to the method for embodiment two, NMR analysis and ESI analysis are carried out, the results showed that the wave spectrum of the ion-pair compound Feature is not the two-part superposition of simple soda acid, and soda acid two parts space length is close, is had an impact, spectral data and original LAE and its hydrochloride are compared, and generate corresponding change, this illustrates to produce between all protons of lauroyl arginine ethyl ester and carbonic acid Strong interaction has been given birth to, and has formd stable single compound structure by strong ionic bond.

Embodiment seven: the measurement of the external minimal inhibitory concentration of lauroyl arginine ethyl ester ion-pair compound (MIC)

Principle and purpose: micro broth dilution method, drug and bacterium according to as defined in CLSI are incubated for for 24 hours altogether in 96 orifice plates Afterwards, the repressed minimum drug concentration of bacterial growth is the minimal inhibitory concentration of the medicine.

Method: lauroyl arginine ethyl ester hydrochloride (LAE) and above-mentioned prepared lauroyl arginine ethyl ester is organic Acid ion to respectively use (TSB) doubling dilution of trypticase soy broth at various concentration, drug and bacterium in 96 orifice plates In mix and be incubated for, separately set abacterial blank control culture medium C K1 and add the culture medium C K2 and not of LAE (1000 μ g/ml) The bacterium normal growth control medium CK3 of drug containing.96 orifice plates are put into be incubated in 37 DEG C of incubators and measure each hole afterwards for 24 hours Absorption photometric value at 625nm.With blank control OD₆₂₅It is worth consistent hole and is considered as bacterium without obvious growth.Bacterium is without obvious growth Drug minimum concentration be LAE to the minimal inhibitory concentration MIC (Minimal Inhibitory Concentration) of bacterium.

The comparison of antibacterial activity of the prepared a variety of LAE derivatives (ion-pair compound) relative to former LAE compound As a result as shown in table 1 below.

The in-vitro antibacterial effect of 1 LAE of table and its ion-pair compound to three kinds of bacteriums

Compare	Escherichia coli	Staphylococcus aureus	Pest of duck Li Moshi bacillus
				LAE	16	8	16
LAE niacin ion pair	16	4	16
				LAE tartaric acid ion pair	16	8	16
LAE oxalic ion pair	8	8	16
				LAE carbonate ions pair	16	16	8

Interpretation of result:

(1) ion-pair compound is most of keeps identical antibacterial activity to Escherichia coli, and especially oxalic ion is to change The antibacterial activity for closing object rises；

(2) ion-pair compound is most of keeps identical antibacterial activity to staphylococcus aureus, and carbonate ions are to change The antibacterial activity decline of object is closed, the antibacterial activity of niacin ion-pair compound significantly rises；

(3) ion-pair compound it is most of to pest of duck Li Moshi bacillus keep identical antibacterial activity, especially carbonic acid from Son obviously rises the antibacterial activity of compound.

Conclusion: the ion-pair compound of LAE derivative will not generate inhibition to the antibacterial activity of the former LAE of single component Effect, is beneficial to antibacterial activity on the contrary.Wherein, niacin ion-pair compound produces staphylococcus aureus significant antibacterial Effect.

Embodiment eight: lauroyl arginine ethyl ester ion-pair compound induces bacterial resistance experiment

Principle and purpose: bacterium can be induced to generate the drug-fast bacteria to the drug under subinhibitory concentration, by long-term The bacterium of subinhibitory concentration drug culture is transferred in fresh pastille culture medium detect the drug-induced bacterium generate it is resistance to The ability of medicine mutation.

Method: the bacterium of logarithmic phase and LAE organic acid ion are mixed in teat glass to solution, and tampon sealing is placed in 37 DEG C, the constant-temperature table culture of 220rpm for 24 hours.Later every the MIC of inspection drug for 24 hours, and muddy bacterium solution will be turned out The bacterium solution of highest drug concentration (the highest drug concentration for being lower than MIC) is added to again with the ratio of 1:100 containing different medicines In the fresh culture of object concentration, 37 DEG C, 220rpm culture for 24 hours, continuing 30 days chloramphenicol is positive controls.After calculating 30 days Ratio of the drug to bacterium MIC and initial MIC.

The ratio of the initial MIC of MIC/ of 2 LAE of table and its ion-pair compound continued stimulus bacterium after 30 days

Compare	Pest of duck Li Moshi bacillus	Escherichia coli	Staphylococcus aureus
				LAE	1	1	2
LAE niacin ion pair	1	1	1
				LAE tartaric acid ion pair	1	1	1
LAE oxalic ion pair	1	1	1
				LAE carbonate ions pair	1	1	1
Florfenicol	8	16	1
				Chloramphenicol	6	12	12
Ampicillin	8	8	2

The results show that LAE organic acid ion can't make bacterium generate tolerance, pest of duck Li Moshi stimulation bacterium 30 days Bacillus and Escherichia coli also do not generate tolerance to LAE, and staphylococcus aureus only generates faint tolerance to LAE single component and supports It is anti-.And the ability that Florfenicol, chloramphenicol, ampicillin induce bacterial resistance is very strong, minimal inhibitory concentration rises within 30 days It is differed to initial 6-16 times.Therefore it is low relative to commercially available antibiotic to induce bacterial resistance ability for LAE and its ion pair, LAE from Son is to the reduced capability for inducing staphylococcus aureus resistance relative to LAE.

Embodiment nine: bacteriosis control efficiency of the lauroyl arginine ethyl ester niacin ion-pair compound to poultry

Oral medication effect of the 1.LAE niacin ion pair to infection riemerella anatipestifer bacillus duckling

Principle and purpose: after duckling bacterial infection, the survival condition of LAE niacin ion pair observation animal is taken orally, LAE is studied Niacin ion pair is to the therapeutic effect for having suffered from bacteriosis poultry.

Method: duckling 50 is only randomly divided into 5 groups, every group 10.By riemerella anatipestifer with 4 × 10⁶The amount of CFU is subcutaneous Injection inoculation is set as infected group four groups of duckling legs, and another group without any processing to be set as blank group.4 infected groups after 12h Respectively in the form of stomach-filling to the LAE aqueous solution of various dose or water blanks control group, upon administration 1h, 7h, 12h, For 24 hours, 48h, 72h, 96h carry out observing and recording death condition respectively, and 96h animal survival condition is shown in Table 3-1, LAE therapeutic effect It is shown in Table 3-2.

Table 3-1: oral therapeutic effect of the LAE niacin ion pair to the duckling for having infected pest of duck Lee Mo Shi bacillus

Table 3-2: the therapeutic effect of oral LAE (referring to earlier application 201711071006.8)

Note: test group is for infection pathogen and to various concentration LAE or the animal of its niacin ion pair；

Blank group is the animal not in contact with pathogenic bacteria；

Control group is infection pathogen but the animal of non-administration LAE or its niacin ion pair.

As the result is shown:

(1) the duckling death rate with Riemerella anatipestifer disease of not oral LAE or its ion pair derivative is 100%；

(2) the illness duckling death rate that 4 milligrams per kilogram of weight LAE niacin ion pairs are taken orally after illness drops to 80%, and The duckling death rate that LAE is administered still is 100%；

(3) taking orally the illness duckling death rate of 8 milligrams per kilogram of weight LAE niacin ion pairs is only 30%, and LAE is administered The duckling death rate be still 60%；

(4) taking orally 16 milligrams per kilogram of weight LAE niacin ion pairs makes the death rate be reduced to 20%, and the young bird of LAE is administered The duck death rate is still 40%；

Conclusion: oral LAE niacin ion pair can treat the Riemerella anatipestifer disease of duckling, and compared to LAEization The effect of object is closed, dosage is lower, and therapeutic effect is more preferable.

Control efficiency of the 2.LAE niacin ion pair to duckling infection riemerella anatipestifer bacillus

Principle and purpose: before bacterial infection, duckling takes orally LAE ion pair, continues oral LAE ion pair daily after infection, Animal survival condition is observed, LAE ion pair is studied as antibacterial agent and protects poultry not by the effect of bacterial invasion.

Method: duckling 50 is only randomly divided into 5 groups, every group 10.Wherein 2 groups are done in the form of stomach-filling to water blanks In addition non-administered group is done experiment group in the form of stomach-filling to LAE niacin ion pair aqueous solution for 3 groups.8h is by duck after being administered for the first time Mo Shi bacillus is in epidemic disease with 4 × 10⁶The amount inoculated with subcutaneous injections of CFU is in three groups of experimental groups and the duckling of one group of non-administration control group Leg is infected group, and another 1 group of non-administered group is without any processing to be uninfected by group (i.e. bacterial infection is not administered as sky yet It is white).Respectively after infection 12h, for 24 hours, 48h to being uninfected by group and infection non-administered group in the form of stomach-filling to water blanks, The LAE niacin ion pair aqueous solution of various concentration is given in 1,2,3 group of experiment respectively.Upon administration 1h, 7h, 12h, for 24 hours, 48h, 72h, 96h carry out observing and recording death condition respectively, and the survival condition of 96h the results are shown in Table 4-1.LAE preventive effect is shown in Table 4- 2。

Table 4-1: oral LAE ion pair derivative resists the control efficiency of pest of duck Lee Mo Shi bacillus infection to duckling.

Table 4-2: oral LAE resists the control efficiency of pest of duck Lee Mo Shi bacillus infection (referring to earlier application to duckling 201711071006.8)

Note: test group is for infection pathogen and to the animal of various concentration LAE or derivatives thereof ion pair；

Blank group is the animal not in contact with pathogenic bacteria and non-administration or its niacin ion pair；

Control group is the animal of infection pathogen but non-administration or its niacin ion pair.

Interpretation of result:

(1) the duckling death rate of the infection pathogen of not oral LAE or its niacin ion pair derivative is 100%；

(2) the illness duckling death rate of oral 4 milligrams per kilogram of weight LAE niacin ion pairs drops to 50% before infection；

(3) the illness duckling death rate of oral 8 milligrams per kilogram of weight LAE niacin ion pairs is only 10% before infection, and The duckling death rate that LAE is administered still is 30%；

(4) the illness duckling death rate of oral 16 milligrams per kilogram of weight LAE is only 20% before infection, not as good as oral 8 milli The control efficiency of grams per kilogram weight LAE niacin ion pair；

(5) the illness duckling death rate of oral 40 milligrams per kilogram of weight LAE niacin ion pairs is still 10% before infection, and The duckling death rate that LAE is administered also is 10%；

Conclusion: it is administered after taking orally LAE niacin ion pair before duckling bacterial infection compared to infection, this intake mode energy It is enough to improve the survival rate of duckling again, and relative to LAE compound, 8 milligrams are the maximum control efficiency for having obtained the latter.Cause The LAE that this oral dose is 4-40mg/kg bw can effectively prevent duckling and infect pest of duck Lee Mo Shi bacillus.Continued administration energy Disease preventing and treating effect is played to a greater degree.10 ducklings of solvent control group are all dead in 96h, and administration group is tied in experiment Still there are 9 survivals when beam, survival rate reaches 90%.In view of 4-8 days treatment cycles and LAE treat the cost of solvent, because It has been the upper limit that production needs that this, which recommends the dosage range of LAE niacin ion pair 10-20mg/kg bw, hence it is evident that lower than LAE's Produce the upper dosage limit range 40mg/kg bw needed.

Therapeutic effect of the 3.LAE niacin ion pair administration to infection Gram-negative bacteria, positive bacteria mouse.

Principle and purpose: orally ingestible and curative effect can be played, is the high requirement to antibacterial agent, therefore we use mouse Domestic animal is represented, respectively with Gram-negative bacteria Escherichia coli come infecting mouse, drug is assessed by oral LAE ion pair later To the therapeutic effect of animal.

Method: 50 Balb/c mouse are randomly divided into 5 groups, every group 10.By the bacterium of logarithmic phase with 10⁸The amount of CFU is infused It is mapped to the abdominal cavity of 4 groups of mouse, another group without any processing to be set as being uninfected by group.1 infected group takes orally the physiology of 0.5ml after 1h Salt water, the other three infected group take orally the LAE ion pair solution of various concentration respectively.Mouse survival situation after counting for 24 hours, obtains To the result of table 5-1, table 6.The Gram-negative bacteria Escherichia coli therapeutic effect of LAE is as shown in table 5-2.

Table 5-1 takes orally the therapeutic effect of LAE niacin ion pair Derivatives In Mice abdominal cavity infection Escherichia coli

Table 5-2: oral LAE is to the therapeutic effect of Murine Model of Intraperitoneal Infection Escherichia coli (referring to earlier application 201711071006.8)

Note:

Test group is the animal of infection pathogen and oral various concentration LAE or its niacin ion pair；

Blank group is the animal not in contact with pathogenic bacteria；

Control group is infection pathogen but non-administered group.

Interpretation of result:

(1) pathogenic bacteria clearance rate in Mice Body is by 0.625 milligrams per kilogram of weight LAE niacin ion pair of administration after infecting 25.6%, effect is better than the LAE with dosage, and the latter's clearance rate is only 21.9%；

(2) pathogenic bacteria clearance rate in Mice Body is by 2.5 milligrams per kilogram of weight LAE niacin ion pairs of administration after infecting 51.2%, effect is better than the LAE with dosage, and the latter's clearance rate is only 44.21%；

(3) pathogenic bacteria clearance rate in Mice Body is by 10 milligrams per kilogram of weight LAE niacin ion pairs of administration after infecting 41.6%, effect is better than the LAE with dosage, and the latter's clearance rate is only 33.95%；

Conclusion: LAE niacin ion pair is administered after mouse infection bacterium compared to administration LAE compound, to intracorporal cause Germ elimination effect is more preferable, and dosage is lower.

According to above-mentioned identical method, it is (golden yellow to infection gram-positive bacteria individually to test the administration of LAE niacin ion pair Staphylococcus) mouse therapeutic effect, the results are shown in Table 6-1.The Gram-negative bacteria Escherichia coli therapeutic effect such as table 6-2 of LAE It is shown.

Table 6-1 takes orally the therapeutic effect of LAE niacin ion pair Derivatives In Mice abdominal cavity infection staphylococcus aureus

Table 6-2 takes orally LAE to the therapeutic effect of Murine Model of Intraperitoneal Infection staphylococcus aureus

Note:

Test group is the animal of infection pathogen and oral various concentration LAE niacin ion pair；

Blank group is the animal not in contact with pathogenic bacteria；

Control group is infection pathogen but non-administered group.

Interpretation of result:

(1) pathogenic bacteria clearance rate in Mice Body is by 2.5 milligrams per kilogram of weight LAE niacin ion pairs of administration after infecting 62.7%, effect is better than the LAE with dosage, and the latter's clearance rate is only 61.5%；

(2) pathogenic bacteria clearance rate in Mice Body is by 10 milligrams per kilogram of weight LAE niacin ion pairs of administration after infecting 60.8%, effect is better than the LAE with dosage, and the latter's clearance rate is only 50.2%；

(3) pathogenic bacteria clearance rate in Mice Body is by 25 milligrams per kilogram of weight LAE niacin ion pairs of administration after infecting 67.2%, effect is better than the LAE with dosage, and the latter's clearance rate is only 61%.

Embodiment ten: lauroyl arginine ethyl ester ion-pair compound is used for the disease of mammal as feed addictive Prevention

By the effective component of the LAE niacin ion pair according to feed total weight, directly it incite somebody to action this with 0.01%~1.0% Product are added in mouse feed；Or this product and carrier are mixed and made into pre-mixing agent；Or with other feed addictives or feedstuff It is mixed and made into premix, concentrate feed form feeding mouse.

6 week old mouse 50 is selected, is randomly divided into 5 processing groups by principle similar in weight.Control group fed corn is small Wheat soybean meal based diets, test group feeding corn wheat bean pulp type and the LAE cigarette for adding feed total weight 0.01%, 0.1%, 1% Acid ion pair, after raising 7 days, disease resistance of the measurement mouse to Escherichia coli.It the results are shown in Table 7-1.Using LAE compound as feeding The feeding effect of feed additives is shown in Table 7-2.

Table 7-1: disease prevention and cure of the feed addition LAE niacin ion pair derivative to mammal

Table 7-2: the feed addition LAE disease prevention and cure to mammal (referring to earlier application 201711071006.8)

Note:

Test group is the feed for contacting pathogenic bacteria and adding various dose LAE or its niacin ion pair；

Blank group is not in contact with pathogenic bacteria；

Control group is the feed for contacting pathogenic bacteria but being not added with LAE or its niacin ion pair

Interpretation of result:

(1) mortality of animals for not launching LAE or the infection pathogen of its niacin ion pair derivative feed is 100%；

(2) contain the feed of 0.01% niacin ion pair derivative in feed, animal survival rate is 60%, is higher than equal conditions The feeding effect 50% of lower LAE；

(3) contain the feed of 0.1% niacin ion pair derivative in feed, animal survival rate is 80%, is higher than equal conditions The feeding effect 70% of lower LAE；

(4) contain the feed of 1% niacin ion pair derivative in feed, animal survival rate is 90%, is higher than under equal conditions The feeding effect 80% of LAE；

Conclusion: from upper table as it can be seen that the mouse survival rate of all test groups compares control group significantly rises, wherein most preferably Group is 3 groups of test, and survival rate 90%, the control group experimental animal for being not added with LAE niacin ion pair is all dead, illustrates LAE Niacin ion pair has preventive and therapeutic effect to mammalian diseases as feed addictive, and relative to LAE compound as feed Additive has better control efficiency.In view for the treatment of cycle and the cost of LAE niacin ion pair therapeutic agent, LAE niacin Ion pair is that 0.01-1%, preferably 0.1-1% can effectively prevent morbidity as the effective concentration of drug or feed addictive, Meet the needs of production.

11: LAE niacin ion pair of embodiment is used for the nutrient growth of mammal as nutrient energy additive

According to the concentration scope of application for the LAE niacin ion pair that embodiment five determines, in the identical base of each group Diet Formula On plinth, 1,2,3 group of test is respectively made an addition to LAE ion pair in perfect compound feed with 0.3%, 0.6%, 0.9% mass ratio, Control group does not add any drug.40 6 week old Balb/c mouse are randomly divided into 4 groups, and each group the 1st is weighed when formal test starts It weight weighed mouse weight after 15 days again.As a result as shown in table 8-1.The result such as table of LAE promotion nutrient growth effect Shown in 8-2.

Table 8-1: influence of the feed addition LAE ion pair derivative to mammalian growth

Table 8-2: the feed addition LAE influence to mammalian growth (referring to earlier application 201711071006.8)

Note:

Test group is to add the feed of various dose LAE and its ion pair；

Control group is the feed for being not added with LAE or its ion pair

Interpretation of result:

(1) the animal average daily gain for not launching LAE or its niacin ion pair derivative feed is almost consistent, and is same Floor level in group test；

(2) daily gain for launching the animal of 0.3%, 0.6%, 0.9% niacin ion pair derivative feed is slightly above thrown Put the level of the LAE feed of same ratio；

It (3) is that 2 groups of levels are apparently higher than 1,3 group in two kinds of feeding trials in terms of average individual net gain in 15 days.

It is indicated above that adding 0.6%LAE niacin ion pair in mouse feed with control group ratio during test, significantly mentioning The average weight gain 12.5% (and the weight gain level of LAE only 9.64%) of high mouse.Therefore, feed addition LAE niacin ion pair energy Enough promote the growth of mammal.

12: LAE niacin ion pair of embodiment is used for the disease prevention and cure of duckling as feed addictive

It is 0.01%~1.0% by the effective component of the LAE niacin ion pair according to feed total weight, directly will This product is added in duckling feed；Or this product and carrier are mixed and made into pre-mixing agent；Or it is former with other feed addictives or feed Material is mixed and made into premix, concentrate feed form feeding duckling.

14 age in days cherry ducklings 50 are selected, are randomly divided into 5 processing groups by principle similar in weight.Each processing sets 1 A repetition (column), 10, every column (half male and half female).Control group fed corn bean pulp type daily ration, test group feed corn bean pulp type simultaneously The LAE niacin ion pair of feed total weight 0.01%, 0.1%, 1% is added, after raising 7 days, measures duckling to Mo Shi in pest of duck Bacillosis disease resistance.It the results are shown in Table 9-1.LAE compound, which is fed, as feed addictive the results are shown in Table 9-2.

Table 9-1: disease prevention and cure effect of the feed addition LAE ion pair derivative to duckling:

Table 9-2: feed addition LAE to the disease prevention and cure effect of duckling (referring to earlier application 201711071006.8)

Note:

Test group is to add the feed of various dose LAE and its niacin ion pair；

Blank group is not in contact with pathogenic bacteria；

Control group is the feed for being not added with LAE or its niacin ion pair

Interpretation of result:

(1) mortality of animals for not launching LAE or the infection pathogen of its niacin ion pair derivative feed is 100%；

(2) contain the feed of 0.01% niacin ion pair derivative in feed, animal survival rate is 70%, same adding conditional The survival rate of the nursing of lower LAE is 60%；

(3) contain the feed of 0.1% niacin ion pair derivative in feed, animal survival rate is 90%, LAE under equal conditions Nursing survival rate be 80%；

(4) contain the feed of 1% niacin ion pair derivative in feed, animal survival rate is LAE under 90%, with equal conditions Feeding effect it is identical；

Conclusion: from upper table as it can be seen that the duckling survival rate of all test groups compares control group and significantly rises, wherein most preferably Group is 3 groups of test, and survival rate 90%, the control group experimental animal for being not added with LAE niacin ion pair is all dead, illustrates LAE Niacin ion pair has preventive and therapeutic effect to duckling disease as feed addictive, it is contemplated that treatment cycle and LAE niacin ion To the cost of therapeutic agent, therefore energy when LAE niacin ion pair as the effective concentration of drug or feed addictive is 0.01-1% Morbidity is effectively prevented, the needs of production are met.

The disease prevention and nutrition of embodiment 13, LAE niacin ion pair as nutrient energy additive for duckling are raw It is long

According to the concentration scope of application for the LAE niacin ion pair that embodiment seven determines, in the identical base of each group Diet Formula On plinth, 1,2,3 group of test is respectively made an addition to LAE ion pair in perfect compound feed with 0.3%, 0.6%, 0.9% mass ratio, Control group does not add any drug.The 1st day weight of groups of animals is weighed when on-test, is weighed weight after 15 days again and is measured Each group is to Riemerella anatipestifer disease disease resistance.As a result as shown in table 10-1.The feeding effect of LAE is shown in Table 10-2.

Table 10-1: disease prevention and cure and nutrient growth effect of the feed addition LAE niacin ion pair derivative to duckling

Table 10-2: feed addition LAE to the disease prevention and cure of duckling and nutrient growth effect (referring to earlier application 201711071006.8)

Note:

Test group is to add not same amount LAE or the feed of its niacin ion pair；

Control group is the feed for being not added with LAE or its niacin ion pair

Interpretation of result:

(1) the animal suffering from disease rate for not launching LAE or the infection pathogen of its niacin ion pair derivative feed is 100%；

(2) containing the feed of 0.3%, 0.6%, 0.9% niacin ion pair derivative in feed, animal suffering from disease rate is 20%, It is identical as the feeding effect of LAE under equal conditions；

(3) the animal average daily gain for not launching LAE or its niacin ion pair derivative feed is almost consistent, and is same Floor level in group test；

(4) daily gain for launching the animal of 0.3%, 0.6%, 0.9% niacin ion pair derivative feed is slightly above thrown Put the level of the LAE feed of same ratio；

It (5) is that 2 groups of levels are apparently higher than 1,3 group in two kinds of feeding trials in terms of average individual net gain in 15 days.

Conclusion: it is indicated above that adding the LAE ion of 0.3%-0.9% in duckling feed with control group ratio during test To the equal average weight gain that can promote duckling, wherein 0.6%LAE ion pair effect of gain is most obvious, significantly mentions compared to control group The average weight gain 11.5% of high duckling.Therefore, it is dead as caused by Riemerella anatipestifer disease both to have can be reduced duckling for LAE ion pair It dies, and duckling growth performance can be improved, promote its nutrient growth.

In view of the cost of growth cycle and LAE ion pair feed addictive, therefore LAE ion pair is as feed addition The effective concentration of agent can effectively prevent morbidity when being 0.3-0.9%, preferably 0.6-0.9%, most preferably 0.6%, meet production It needs.

Disease prevention and battalion of 14: the LAE niacin ion pair of embodiment as nutrient energy additive for aquatic livestock Health is long

Selecting weight is that the 240 tail Tilapia mossambicas of 40.9 ± 0.18g are divided into 4 processing groups, adds different graded levels respectively LAE niacin ion pair, every group of 3 repetitions, every 20 tail fish of repetition.By 8 weeks growth tests, to evaluate LAE niacin ion To the influence to Growth Op Tilapia performance.It raises simultaneously, to probe into LAE niacin ion pair to aquatic livestock bacteria resistance disease Ability carries out Aeromonas sobria infection containing LAE niacin ion pair 8 weeks Tilapia mossambicas of feed for feeding, with Rofe after infection The survival rate of fish embodies its premunition.It the results are shown in Table 11-1.The effect for feeding LAE is shown in Table 11-2.

Influence of the table 11-1 feed addition LAE niacin ion pair derivative to Tilapia mossambica nutrient growth

Influence of the table 11-2 feed addition LAE to Tilapia mossambica nutrient growth (referring to earlier application 201711071006.8)

Note:

Test group is to add the feed of various dose LAE or its niacin ion pair；

Control group is the feed for being not added with LAE or its niacin ion pair

Interpretation of result:

(1) animal survival rate for not launching LAE or the infection pathogen of its niacin ion pair derivative feed is 0%；

(2) contain the feed of 0.01% niacin ion pair derivative in feed, animal survival rate is 63.66%, and opposite increasing Rate is 250.1% again, is above the nursing survival rate and body weight increase rate of LAE under equal conditions；

(3) contain the feed of 0.1% niacin ion pair derivative in feed, animal survival rate is 80%, and body weight increase rate It is 258.3%, is above the nursing survival rate and body weight increase rate of LAE under equal conditions；

(4) contain the feed of 1% niacin ion pair derivative in feed, animal survival rate is 63.66%, and body weight increase rate It is 250.1%, is above the nursing survival rate and body weight increase rate of LAE under equal conditions；

It (5) is that 2 groups of levels are apparently higher than 1,3 group in two kinds of feeding trials in terms of average individual net gain in 8 weeks.Knot By: adding LAE niacin ion pair in the feed of Tilapia mossambica can be improved 3.52%, 6.91%, 4.64% and of body weight increase rate Specific growth rate 7.94%, 13.08%, 6.54% reduces feed coefficient 4.76%, 5.56%, 2.38%, promotes Tilapia mossambica Growth.

(6) although in two groups of tests, the survival rate highest of 1% dosage group, growth performance is not highest, but 0.1% Dosage group.From the point of view of the practice for the treatment of, a certain range of dosage of drug, which increases, can increase effect of curing the disease, but too high dose Administration will affect the growth of host sometimes, traces it to its cause and still requires study, it may be possible to which too high dose has secondary make to nutrient growth With.

(7) anti-infective experimental result is shown, the test fish for eating the not ion pair of niacin containing LAE feed is all dead, is eaten Tilapia mossambica survival rate containing LAE niacin ion pair is respectively 63.66%, 80%, 91.67%.Illustrate to add 0.01%-1% LAE ion pair is able to ascend the anti-infection ability of fish really.

In view of the cost of growth cycle and LAE niacin ion pair feed addictive, therefore LAE niacin ion pair conduct The effective concentration of feed addictive can effectively prevent morbidity when being 0.01-1%, preferably 0.1-1%, most preferably 0.1%, meet life The needs of production.

Embodiment 15, LAE ion pair are compared with antibiotic, LAE are to the therapeutic effect of diarrhea piglet

Applicant entrusts the large-scale pig farm in Henan Province, is administered SY to 118 piglets for diarrhea occur on its farm The experiment of (i.e. LAE), LAE ion pair.

Another setting gentamicin administration group is as parallel control.

Daily early (7:00), in (12:00), late (18:00) 3 times, perfusion is carried out to sick pig.

Evaluation criterion: diarrhea is not necessarily to the animal of medication after curing, and is calculated as the animal of remaining survival, i.e. test stops；

Invalid dead animal, is calculated as dead animal after piglet medication, i.e. test stops.

As a result as shown in table 12.

Table 12

As shown above, using the piglet of gentamicin treatment, average cure rate is 77.5%.However, the dosage is Considerably beyond the dosage of Conventional antibiotic, imply that, in order to obtain the cure rate, the intracorporal antibiotic residue of piglet will be shown It writes exceeded.

The piglet treated using LAE, the cure rate of minimum dose 10mg/kg are 72.2%, and maximum dose 50mg/kg's controls More rate is 78.57%, and considerably beyond the cure rate of the antibiotic of same dose, (cure rate of the gentamicin of 10mg/kg is not Display), this shows LAE on treatment grice diarrhoea, and compared to gentamicin, the cure rate of identical dosage is higher, and Because of its good biological metabolism characteristic, it will indicate the residual that any hazardous drugs are not present in piglet body, to substitute antibiosis for animals Element provides good direction.

It is in progress in the test for the treatment of diarrhea piglet as LAE ion pair.But according to embodiment nine to embodiment ten The LAE ion-pair compound unanimously shown in four is better than the test result of LAE compound in antibiotic property, and inventor is expected LAE ion-pair compound will be better than the therapeutic effect of LAE compound in the result for the treatment of diarrhea piglet, this imply that LAE ion Antibiotic can equally be replaced to compound and be widely used in agricultural production.

Claims (21)

1. lauroyl arginine ethyl ester (LAE) derivative with the structural formula as shown in following formula (I):

Wherein,

X is organic acid RCOO^-, it is selected from salicylic acid, formic acid, acetic acid, diacetic acid, propionic acid, propionic acid, butyric acid, lactic acid, benzoic acid, mountain The arbitrary one or more of pears acid, fumaric acid, citric acid, tartaric acid, malic acid, phosphoric acid, oxalic acid or carbonic acid；

R₁It is the linear saturated fatty acids group containing 8-14 carbon atom or the oxygen-containing acidic group of straight chain containing 8-14 carbon atom Group；

R₂It is straight chain fatty acid groups containing 1-18 carbon atom or the Branched fatty acid groups containing 1-18 carbon atom or contains There are the aromatic group of 1-18 carbon atom, or the straight chained alkyl containing 1-4 carbon atom；

R₃It is one kind of having structure:

-NH₃

The range of n is 0-4.

2. derivative as described in claim 1, wherein the derivative is the lauroyl with structural formula shown in following (III) Arginine ethyl ester organic acid ion is to compound:

Wherein, RCOO^-For the acid group of acid as described below: being selected from salicylic acid, formic acid, acetic acid, diacetic acid, propionic acid, propionic acid, butyric acid, cream Acid, benzoic acid, sorbic acid, fumaric acid, citric acid, tartaric acid, malic acid, phosphoric acid, oxalic acid or carbonic acid or niacin, tartaric acid, grass Acid.

3. derivative as claimed in claim 1 or 2 is used to prepare the pathogenic infection for treating or preventing poultry, aquatic livestock The purposes of drug.

4. purposes as claimed in claim 3, wherein the drug is for treating or preventing duckling Infective gram The drug of negative bacterium, and the drug with every kg body weight give the dosage of derivative shown in 4-40mg formula (I) or (III) into Row application.

5. purposes as claimed in claim 4, wherein being given with every kg body weight derivative shown in 8-16mg formula (I) or (III) The dosage of object is administered, and the Pathogenic gram-negative bacterium is riemerella anatipestifer.

6. purposes as claimed in claim 5, wherein giving derivative shown in 16mg formula (I) or (III) with every kg body weight Dosage be administered, with treat duckling infection riemerella anatipestifer bacillus disease；Or it gives shown in 8mg formula (I) or (III) The dosage of derivative is administered, to prevent the disease of duckling infection riemerella anatipestifer bacillus.

7. purposes as claimed in claim 3, wherein the drug is for treating or preventing the pathogenic of poultry, aquatic livestock The drug of Gram negative bacterial disease, and derivative shown in 0.625-10mg formula (I) or (III) is given with every kg body weight Dosage be administered.

8. purposes as claimed in claim 7, wherein the dosage is 2.5-10mg, in the Pathogenic gram-negative bacterium is Mo Shi bacillus, Escherichia coli, Pasteurella, salmonella, haemophilus, brucella or Aeromonas sobria.

9. purposes as claimed in claim 3, wherein the drug is the gram for treating or preventing poultry, aquatic livestock The drug of positive bacterial diseases, and the drug gives derivative shown in 2.5-25mg formula (I) or (III) with every kg body weight Dosage is administered.

10. purposes as claimed in claim 9, wherein the gram-positive bacteria is staphylococcus aureus, streptococcus, erysipelas Bacillus, mycobacteria, bacillus anthracis, and the drug is given shown in 2.5,10 or 25mg formula (I) or (III) with every kg body weight The dosage of derivative is administered.

11. such as the described in any item purposes of claim 4-10, wherein the poultry, aquatic livestock include: chicken and duck goose turkey quail Quail pigeon, pig, cattle and sheep horse camel cat and dog and fish shrimp crab, disease caused by the pathogenic bacteria includes yellowish-white dysentery, asthma, erysipelas Disease, oedema, clostridium property enteritis, Proliferative Enteritis, tuberculosis, pasteurellosis, bacillus anthracis disease, salmonellosis.

12. purposes as claimed in claim 3, wherein the drug is the drug for treating grice diarrhoea, and the drug Piglet is administered with the dosage that every kg body weight gives derivative shown in 10-50mg formula (I) or (III).

13. purposes as claimed in claim 12, wherein the drug gives 30-50mg formula (I) or (III) with every kg body weight Shown in the dosage of derivative be administered piglet.

14. purposes as claimed in claim 12, wherein the drug gives 50mg formula (I) or (III) institute with every kg body weight The dosage of the derivative shown is administered piglet.

15. derivative as claimed in claim 1 or 2 or such as described in any item purposes of claim 4-10, wherein be suitble to formula (I) or the pharmaceutical dosage form of derivative shown in (III) includes granule, solution, suspending agent, pulvis, capsule, finish, paste.

16. derivative as claimed in claim 1 or 2 is in preparation for changing answering in the polar drug of cell membrane of microorganism With.

17. the derivative as shown in formula (I) in claims 1 or 2 or (III) is for changing the polar side of cell membrane of microorganism Method, step include:

(1) invasive organism solution is diluted to OD600=0.05, cell membrane polar dies is then added；

(2) derivative solution shown in formula (I) or (III) is added until final concentration of 16 μ g/ml, sufficiently reacts；

(3) using the light of 622nm wavelength as exciting light, the fluorescent value at 670nm is measured；

(4) fluorescent value for passing through each sample of flow cytomery, show that bacterial cell membrane goes to pole by ratio calculated The degree of change.

18. the preparation method of lauroyl arginine ethyl ester (LAE) derivative shown in formula (III), step include:

(1) formula (II) compound represented is dissolved by heating, acylate solution is then added；

(2) under heating conditions, lauroyl arginine ethyl ester ion pair chemical combination shown in formula (III) is generated by following reaction equations Object；

19. method as claimed in claim 18, wherein the organic acid RCOO^-Acid group selected from following acid: salicylic acid, first Acid, acetic acid, sodium Diacetate, propionic acid, butyric acid, lactic acid, benzoic acid, sorbic acid, fumaric acid, citric acid, tartaric acid, malic acid, phosphorus Acid, oxalic acid or carbonic acid；And selected from the acid group with the niacin for animal extra-nutrition energy, pantothenic acid, folic acid, ascorbic acid.

20. method as claimed in claim 19, wherein the organic acid is selected from niacin, tartaric acid, oxalic acid, carbonic acid.

21. wherein the organic acid is added in methanol solution, and adding such as described in any item methods of claim 17-20 Enter suitable NaOH, be stirred at room temperature until white solid is precipitated, suction filtration is simultaneously washed with methanol point, obtains acylate.