CN108218739B

CN108218739B - Butyrylglutamic acid derivative, and composition and application thereof

Info

Publication number: CN108218739B
Application number: CN201711397106.XA
Authority: CN
Inventors: 彭险峰
Original assignee: Guangzhou Insighter Biotechnology Co Ltd
Current assignee: Guangzhou Insighter Biotechnology Co Ltd
Priority date: 2017-12-21
Filing date: 2017-12-21
Publication date: 2020-05-01
Anticipated expiration: 2037-12-21
Also published as: CN108218739A

Abstract

The invention discloses a butyrylglutamic acid derivative, a composition and application thereof. The butyrylglutamic acid derivative, or racemate, stereoisomer, geometric isomer, tautomer, solvate or feed acceptable salt thereof can be applied to preparation of novel feed additives and preparation of feeds. The invention also discloses a feeding composition of the butyrylglutamic acid derivative, or a racemate, a stereoisomer, a geometric isomer, a tautomer, a solvate or a feed acceptable salt thereof. The butyrylglutamic acid derivative provided by the invention has the improvement effects of increasing the animal weight gain rate, reducing the feed conversion ratio, controlling the diarrhea rate and other animal production performances when being applied to the animal breeding industry, and can be used as an effective and safe novel feed additive.

Description

Butyrylglutamic acid derivative, and composition and application thereof

The technical field is as follows:

the invention belongs to the field of animal feed additives, and particularly relates to a butyrylglutamic acid derivative, a composition containing the butyrylglutamic acid derivative, and application of the butyrylglutamic acid derivative and the composition containing the butyrylglutamic acid derivative in preparation of an animal feed additive or feed.

Background art:

the feed is a product for animals after industrial processing and manufacturing, and is a main animal food in modern intensive breeding industry. In the process of animal breeding, farmers desire that the feed products eaten by animals can be absorbed and utilized by the animals to the maximum extent, and the production performance of the animals is improved, so that the aim of improving the economic benefit of breeding is fulfilled. The feed additive with corresponding effect is added into the feed product, so that the utilization rate of the feed product can be effectively improved.

The feed additive is a small amount or trace substances added in the processes of processing, preparing and using the feed, and comprises nutritional feed additives and common feed additives. A nutritive feed additive is a small amount of trace substances, including feed-class amino acids, vitamins, minerals, trace elements, enzyme preparations, non-protein nitrogen, etc, which are added in feed for supplementing the nutrients of feed. The general feed additive refers to a small amount or trace amount of substances which are added into the feed to ensure or improve the quality of the feed and improve the utilization rate of the feed. The common feed additives commonly used in the field at present and having the functions of efficiently and stably improving the feed utilization rate and improving the animal production performance mainly comprise a high-dose copper agent, a high-dose zinc agent, feed antibiotics, a chemical synthetic antibacterial agent and the like, but the substances have large side effects in the breeding industry after long-term use, such as the defects of liver and kidney toxicity, growth inhibition, kidney function damage, urinary tract disorder, teratogenesis, mutagenesis, drug resistance generation, drug residue, environment pollution and the like of animals. The search for new feed additives that are effective and safe in order to safeguard the health of animals and to improve the productivity of the breeding industry is an urgent problem to be solved in the art.

The invention content is as follows:

based on the butyrylglutamic acid derivative, racemate, stereoisomer, geometric isomer, tautomer, solvate or feed acceptable salt thereof, the butyrylglutamic acid derivative and related substances thereof can be used for improving the production performance of animals and can be applied to the preparation of animal feed additives or animal feeds; the invention also provides a feeding composition containing the butyrylglutamic acid derivative or racemate, the stereoisomer, geometric isomer, tautomer, solvate or feed acceptable salt thereof, and the composition can be used for preparing animal feed additives or preparing animal feeds or being used as the animal feeds in animal breeding industry.

In one aspect, the invention provides a butyrylglutamic acid derivative with a structure shown as a formula (I), or a racemate, a stereoisomer, a geometric isomer, a tautomer, a solvate or a feed acceptable salt thereof:

wherein, the substituent groups A and B in the formula are respectively H or alkyl, and the alkyl is n-propyl, isopropyl or C₅-C₂₀Straight or branched alkyl, C₁-C₈Straight or branched haloalkyl or C₃-C₇A cycloalkyl group.

In some embodiments, the substituent groups a and B of the butyrylglutamic acid derivative are both alkyl.

In some embodiments, the substituent groups a and B of the butyrylglutamic acid derivative are not both H.

In another aspect, the invention provides an application of the butyrylglutamic acid derivative provided by the invention, or a racemate, a stereoisomer, a geometric isomer, a tautomer, a solvate or a feed acceptable salt thereof in preparing an animal feed additive.

In some embodiments, the farmed animals to which the animal feed additive is applied are selected from livestock, poultry, aquaculture animals or pets.

In another aspect, the invention also provides an application of the butyrylglutamic acid derivative provided by the invention, or a racemate, a stereoisomer, a geometric isomer, a tautomer, a solvate or a feed acceptable salt thereof in preparing animal feed.

In another aspect, the invention further provides a feeding composition, which comprises at least one of the butyrylglutamic acid derivatives provided by the invention, or racemate, stereoisomer, geometric isomer, tautomer, solvate or feed acceptable salt thereof, and a feeding auxiliary material.

Optionally, the feedable adjuvant is selected from a feedable carrier, a diluent, an excipient, a vehicle or a combination thereof.

In some embodiments, the feed composition further comprises feed materials and or additional animal feed additives.

Optionally, the additional animal feed additive is selected from a nutritional feed additive, a general feed additive or a pharmaceutical feed additive.

In another aspect, the invention provides a method for improving the production performance of farmed animals.

In some embodiments, the method comprises administering the butyrylglutamic acid derivatives provided herein, or racemates, stereoisomers, geometric isomers, tautomers, solvates or feed acceptable salts thereof, to a farm animal.

In some embodiments, the method comprises administering to a farm animal a feeding composition provided by the present invention comprising a butyrylglutamic acid derivative provided by the present invention, or a racemate, a stereoisomer, a geometric isomer, a tautomer, a solvate, or a feed-acceptable salt thereof.

In another aspect, the present invention also provides a method for preparing, separating and purifying the butyrylglutamic acid derivative or its racemate, stereoisomer, geometric isomer, tautomer, solvate or feed-acceptable salt.

The invention has the beneficial effects that:

the invention discovers that the butyrylglutamic acid and the derivative thereof have the effects of improving the production performance such as increasing the average daily gain, reducing the feed conversion ratio and the like on livestock, poultry or aquaculture animals, and can also effectively prevent and treat the diarrhea state of the livestock, the poultry or pets. The butyrylglutamic acid prodrug ester compound provided by the invention is found to have more remarkable effect on improving the production performance of animals in screening butyrylglutamic acid derivatives which can be used as novel feed additives, probably because the increase of the oil-water distribution coefficient of the butyrylglutamic acid prodrug ester compound improves the absorption performance of cultured animals on the butyrylglutamic acid derivatives, and meanwhile, the butyrylglutamic acid derivative can be used as a miniature storage base to improve the bioavailability by slowly hydrolyzing the butyrylglutamic acid derivative in the animal body for slowly releasing the butyrylglutamic acid.

Any embodiment of any aspect of the invention may be combined with other embodiments as long as there is no conflict between them. Furthermore, in any embodiment of any aspect of the present invention, any technical feature may be applied to that technical feature in other embodiments as long as there is no contradiction therebetween.

The foregoing merely summarizes certain aspects of the invention and is not intended to be limiting. The foregoing and other aspects are more fully described below.

Further details of the invention are described.

Reference will now be made in detail to certain embodiments of the invention, examples of which are illustrated by the accompanying structural and chemical formulas. The invention is intended to cover alternatives, modifications and equivalents, which may be included within the scope of the invention as defined by the appended claims. Furthermore, certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment or in any suitable subcombination.

A compound is provided.

The structure of the butyrylglutamic acid derivative provided by the invention is shown as the formula (I):

in the formula, the substituent groups A and B are respectively H or alkyl, and the alkyl is n-propyl, isopropyl or C₅-C₂₀Straight or branched alkyl, C₁-C₈Straight or branched haloalkyl or C₃-C₇A cycloalkyl group.

In general, the term "substituted" means that one or more hydrogen atoms in a given structure is replaced with a particular substituent. Unless otherwise indicated, a substituted group may have one or more substituents substituted at each substitutable position of the group. When more than one position in a given formula can be substituted with one or more substituents selected from a particular group, the substituents may be substituted at each position, identically or differently.

Some of the chemical groups involved in the present invention express: "C₅-C₂₀Straight-chain or branched alkyl "represents a straight-chain or branched alkyl group having 5 to 20 carbon atoms; "C₁-C₈Straight-chain or branched haloalkyl "represents a straight-chain or branched alkyl hydrocarbon radical containing from 1 to 8 carbon atoms, in which one or more hydrogen atoms are replaced by a halogen selected from fluorine, chlorine, bromine or iodine; "C₃-C₇Cycloalkyl "stands for cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

The preparation method of the butyrylglutamic acid derivative comprises the following steps: step 1, preparing glutamate from glutamic acid and alcohol in an organic reaction solvent under the catalysis of thionyl chloride or concentrated sulfuric acid; and 2, carrying out amidation reaction on the glutamate and butyryl chloride under the alkaline condition of a dry organic reaction solvent system or carrying out amidation reaction on the glutamate and butyric anhydride under the acidic condition to generate corresponding butyryl glutamate.

In some embodiments, step 1 of the method for preparing butyrylglutamic acid derivatives is to perform an amidation reaction of glutamic acid and butyrylchloride under the alkaline condition of a dry organic reaction solvent system to generate butyrylglutamic acid diester, and step 2 of the method for preparing butyrylglutamic acid derivatives is to perform an amidation reaction of glutamic acid and alcohol in an organic reaction solvent system under the catalysis of thionyl chloride to generate glutamate.

Optionally, the alcohol is n-propanol, isopropanol, C₅-C₂₀Straight or branched chain alkanol, C₁-C₈Straight or branched halogenoalkanols or C₃-C₇Cycloalkanol, the organic reaction solvent involved is a polar organic solvent.

Further, the polar organic solvent includes an alcoholic organic solvent or an aprotic organic solvent.

In particular, the organic reaction solvent involved in step 1 includes, but is not limited to, the short-chain fatty alcohol, methyl tertiary butyl ether, tetrahydrofuran, N-methylpyrrolidone, toluene, dichloromethane, N-dimethylformamide or N, N-dimethylacetamide involved.

In some embodiments, the organic reaction vehicle involved in the acylation of glutamate with butyryl chloride includes, but is not limited to, tetrahydrofuran, N-methylpyrrolidone, toluene, methylene chloride, dimethyl sulfoxide, N-dimethylformamide, or N, N-dimethylacetamide.

In some embodiments, step 1 of the method for preparing butyrylglutamic acid derivatives is to prepare glutamic acid monoester with alcohol in an organic reaction medium under catalysis of concentrated sulfuric acid; step 2, carrying out amidation reaction on the glutamate and butyric anhydride under an acidic condition to generate corresponding butyrylglutamic acid monoester.

Optionally, the alcohol is n-propanol, isopropanol, C₅-C₂₀Straight or branched chain alkanol, C₁-C₈Straight or branched halogenoalkanols or C₃-C₇Cycloalkanol, toThe organic reaction solvent is polar organic solvent.

Specifically, the organic reaction solvent involved in the reaction of step 1 to form the ester includes, but is not limited to, short-chain fatty alcohol, methyl tertiary butyl ether, tetrahydrofuran, N-methylpyrrolidone, toluene, dichloromethane, N-dimethylformamide or N, N-dimethylacetamide.

Optionally, the acidic condition is an organic acid or an inorganic acid.

Further, the organic acid includes, but is not limited to, formic acid, acetic acid, propionic acid, butyric acid, trifluoroacetic acid, methanesulfonic acid, or ethanesulfonic acid.

Further, the organic acid includes, but is not limited to, hydrochloric acid, sulfuric acid, phosphoric acid, or nitric acid.

In other embodiments, the method for preparing the butyrylglutamic acid derivative involves a change from glutamic acid to butyrylglutamic acid monoester according to the following formula (II):

a is n-propyl, isopropyl or C₅-C₂₀Straight or branched alkyl, C₁-C₈Straight or branched haloalkyl or C₃-C₇A cycloalkyl group.

In other embodiments, the method for preparing the butyrylglutamic acid derivative involves a course of change from glutamic acid to butyrylglutamic acid monoester as follows:

In some embodiments, the present invention also provides a preparation method of butyrylglutamic acid derivative as butyrylglutamic acid mixed ester, the preparation method comprises the steps of 1, preparing glutamic acid monoester by glutamic acid and alcohol in organic reaction solvent under the catalysis of concentrated sulfuric acid; step 2, glutamic acid monoester and alcohol in an organic reaction solvent are catalyzed by thionyl chloride to generate glutamic acid mixed ester; and 3, carrying out amidation reaction on the glutamate and butyryl chloride under the alkaline condition of a dry organic reaction solvent system or carrying out amidation reaction on the glutamate and butyric anhydride under the acidic condition to generate corresponding butyryl glutamate.

The "mixed ester" according to the present invention means an ester compound produced by esterification of a carboxyl group of a polybasic fatty acid with a different alcohol.

Specifically, the organic reaction solvent involved in the reaction to form the ester in step 1 includes, but is not limited to, short-chain fatty alcohol, methyl tert-butyl ether, tetrahydrofuran, N-methylpyrrolidone, toluene, dichloromethane, N-dimethylformamide or N, N-dimethylacetamide, and the organic reaction solvent involved in the reaction of glutamate with butyryl chloride to produce an acylation reaction includes, but is not limited to, tetrahydrofuran, N-methylpyrrolidone, toluene, dichloromethane, dimethyl sulfoxide, N-dimethylformamide or N, N-dimethylacetamide.

Optionally, the acidic condition is an organic acid or an inorganic acid.

In some embodiments, the glutamic acid is a chiral compound, and the glutamic acid of the present invention is selected from L- (-) -glutamic acid (structure formula (IV)), D- (+) -glutamic acid (structure formula (V)), or DL- (±) glutamic acid as racemate, and the reaction of the L- (-) -glutamic acid and butyryl chloride gives a stereoisomer or racemate of a butyrylglutamic acid derivative having a chiral center.

In some embodiments, the chiral stereoisomer of glutamic acid and the stereoisomer of butyrylglutamic acid derivative can undergo a stereoconfigurational transformation under suitable conditions, such as a stereoconfigurational interconversion of glutamic acid or butyrylglutamic acid derivative, as follows:

when the related alcohol reacts with glutamic acid and the like to generate the corresponding butyrylglutamic acid derivative with a rigid structure, different geometric isomer products can be generated during the reaction of the reaction substrate.

The stereoisomers, geometric isomers and tautomers described above are also included in the practice of the present invention.

In some embodiments, the present invention provides a process for preparing a butyrylglutamic acid derivative, further involving a process for separating, purifying or recrystallizing the reaction product. The reaction product can be used for obtaining crude products from the reaction system by a solvent removal method. In order to obtain a solid substance with higher chemical purity and lower impurity content, the crude product is dissolved, crystallized or precipitated or recrystallized and separated in an alcohol solvent, an alcohol-water mixed solvent or other organic solvents which can be used for product recrystallization under the conditions of proper temperature, illumination, mechanical vibration and the like to obtain the butyrylglutamic acid derivative with a certain crystal form state. The butyrylglutamic acid derivative having a certain crystal form state is a crystal of the butyrylglutamic acid derivative or a solvate of the butyrylglutamic acid derivative. The solvate of the butyrylglutamic acid derivative may be selected from a hydrate of the butyrylglutamic acid derivative or an ethanolate of the butyrylglutamic acid derivative.

The term "solvate" as used herein refers to a eutectic association formed by binding a stoichiometric or non-stoichiometric amount of solvent molecules by non-covalent intermolecular forces due to external and internal conditions during the contact between the compound of the present invention and the solvent molecules. Solvents that form solvates include, but are not limited to, water, acetone, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, isopropanol, and the like. "hydrate" refers to an association or crystal of solvent molecules with water, i.e., a compound that binds a stoichiometric or non-stoichiometric amount of water by non-covalent intermolecular forces.

The preparation of the butyrylglutamic acid derivative provided by the invention can be used for obtaining solid substances with higher chemical purity and lower impurity content and can be processed after salting out. The salting-out method is a process of performing salt formation and precipitation on the butyrylglutamic acid derivative and corresponding organic base, inorganic base, organic acid or inorganic acid by utilizing the principles of an acid-base neutralization method, an acid-base coordination method or an acid-base chelation method to obtain a salt acceptable for the feed.

Specifically, the feed acceptable salt is a salt formed by the butyrylglutamic acid derivative and an organic base, an inorganic base, an organic acid or an inorganic acid which is nontoxic to animals. By "feed acceptable" is meant that the substance or composition must be chemically or toxicologically compatible in connection with the constituent feed or the edible farmed animals.

In some embodiments, the butyroglutamic acid derivative is a di-or mixed ester of butyroglutamic acid, and the post-treatment salting-out precipitation process forms an acid-base complex salt and or an acid-base chelate salt with an inorganic or organic acid, including but not limited to acetate, maleate, succinate, mandelate, fumarate, malonate, malate, 2-hydroxypropionate, pyruvate, oxalate, glycolate, salicylate, glucuronate, galacturonate, citrate, tartrate, aspartate, glutamate, benzoate, p-methylbenzoate, cinnamate, p-toluenesulfonate, benzenesulfonate, methanesulfonate, ethanesulfonate, trifluoromethanesulfonate, or a combination thereof; the inorganic acid includes, but is not limited to, hydrochloride, hydrobromide, phosphate, sulfate, nitrate, or combinations thereof.

In some embodiments, the butyrylglutamic acid derivative is a monoester of butyrylglutamic acid, and the post-treatment salting-out precipitation process forms an acid-base coordination salt and/or an acid-base chelate salt with an organic or inorganic acid, or forms an acid salt with an organic or inorganic base. The organic acid includes, but is not limited to, acetate, maleate, succinate, mandelate, fumarate, malonate, malate, 2-hydroxypropionate, pyruvate, oxalate, glycolate, salicylate, glucuronate, galacturonate, citrate, tartrate, aspartate, glutamate, benzoate, p-methylbenzoate, cinnamate, p-toluenesulfonate, benzenesulfonate, methanesulfonate, ethanesulfonate, trifluoromethanesulfonate or a combination thereof; the inorganic acid includes, but is not limited to, hydrochloride, hydrobromide, phosphate, sulfate, nitrate, or combinations thereof. The organic base includes but is not limited to ammonia or triethylamine. The inorganic base includes, but is not limited to, sodium hydroxide, potassium hydroxide, or calcium hydroxide.

The butyrylglutamic acid derivative provided by the invention, or racemate, stereoisomer, geometric isomer, tautomer, solvate or feed acceptable salt thereof has one of the following structures, but is not limited to the following structure:

the invention relates to application of a butyrylglutamic acid derivative.

The butyrylglutamic acid is easily soluble acylated glutamic acid, and the invention finds that the butyrylglutamic acid can obviously improve the average daily gain, reduce the feed conversion rate and effectively prevent and treat diarrhea symptoms of animals when being singly used for animal breeding in the breeding test of the animals for the first time. The inventor finds out in the metabolic dynamics research that the metabolic pathway of butyrylglutamic acid in an animal body is mainly to be hydrolyzed into butyric acid and glutamic acid without discharging the protoform out of the body. Butyric acid is one of the components of the excreta of the organism, and glutamic acid is an important substance of protein metabolism in the organism to participate in a plurality of important chemical reactions, so that it can be seen that butyrylglutamic acid can be used as a safe and effective novel animal feed additive.

The butyrylglutamic acid derivative is a prodrug compound of butyrylglutamic acid. The "prodrug compound" represents a butyrylglutamic acid which is converted into water-soluble butyrylglutamic acid in vivo, and such conversion is influenced by the prodrug compound in the gastrointestinal tract, blood or tissue fluid in vivo via an enzyme or the pH value in vivo. The prodrug compound of butyrylglutamic acid provided by the invention can obviously improve the oil-water distribution coefficient of the butyrylglutamic acid derivative, increase the lipid solubility, and improve the absorption performance of organisms, thereby improving the bioavailability.

Therefore, the butyrylglutamic acid derivative can be used in the preparation of an animal feed additive or animal feed.

The term "animal" as used herein means a human or a cultured animal which is incapable of synthesizing organic matter from inorganic matter and capable of having life activities such as feeding, digestion, absorption, respiration, circulation, excretion, sensation, exercise, and reproduction by using only organic matter as a food. "farmed animals" include poultry, livestock, aquaculture animals, and other animals that are artificially reared to be legally caught, including pets, such as cats and dogs. The term "livestock" is, for example, any of pigs, cattle, horses, goats, sheep, deer, and many useful rodents. The term "poultry" is intended to include, for example, chickens, ducks, geese, quail, pigeons and the like. The term "aquaculture animal" includes, for example, fish, shrimp, turtles, and the like.

The present invention also provides a method for improving the productivity of farmed animals, which comprises mixing butyrylglutamic acid derivatives, or racemates, stereoisomers, geometric isomers, tautomers, solvates or feed acceptable salts thereof with animal feed, and then feeding the animals.

In some breeding schemes, the butyrylglutamic acid derivative is fed to poultry, so that the utilization rate of feed can be improved, the feed conversion ratio coefficient is obviously reduced, and the diarrhea rate of the poultry can be effectively controlled.

Optionally, the butyrylglutamic acid derivative is fed to poultry such as chickens, ducks, geese or pigeons for eating, the utilization rate of poultry feed is improved, the feed conversion ratio coefficient is obviously reduced, and the diarrhea rate of the poultry can be effectively controlled.

In some breeding schemes, the butyrylglutamic acid derivative is fed to livestock, so that the weight gain of the livestock can be obviously improved, the feed conversion rate of the livestock can be obviously reduced, and the feed conversion rate of the livestock can be improved.

Optionally, the butyrylglutamic acid derivative can be eaten by monogastric or ruminant domestic animals such as pigs, cows and sheep, and can effectively improve the conversion rate of the domestic animal feed.

In some breeding programs, the butyrylglutamic acid derivatives are used in combination with aquaculture feed, and the feed factor of the aquatic animals is improved to various degrees.

Optionally, the butyrylglutamic acid derivative is matched with fish feed, shrimp feed and the like, so that the feed coefficient is reduced to different degrees.

In some breeding schemes, the butyrylglutamic acid derivative can be used together with pet food such as cat food and dog food, so that the effect of regulating the gastrointestinal function of pets such as cats and dogs is achieved, and the diarrhea symptom of the pets caused by dyspepsia is effectively relieved.

The invention relates to a feed additive, which is a small amount or trace substance added in the processes of processing, manufacturing and using feed and is divided into nutritional feed additives and common feed additives which are also called non-nutritional feed additives. The nutritive feed additive is a small amount or trace substances added into compound feed to balance feed nutrients, improve feed utilization rate and directly exert a nutritive effect on animals, and comprises vitamins, trace elements, amino acids, small peptides and non-protein nitrogen. The general feed additive is also called non-nutritive additive, and refers to some non-nutritive substances which are added into the feed for improving the utilization rate of the feed, ensuring the quality and quality of the feed and being beneficial to the health or metabolism of animals, and comprises growth promoters, insect repellent health care agents, feed modifying agents, feed modulating agents, feed storing agents and Chinese herbal medicine additives.

The feed is a product which is industrially processed and manufactured and is eaten by animals.

The invention provides a feeding composition, which comprises one or more of butyrylglutamic acid derivatives provided by the invention, or racemates, stereoisomers, geometric isomers, tautomers, solvates or feed-acceptable salts thereof and a feeding auxiliary material.

The composition refers to a compound set comprising one or more compounds as active ingredients.

The term "comprising" as used herein is open-ended, meaning that the meaning of the term is included, but not exclusive of other aspects.

Optionally, the feedable auxiliary materials comprise feed additives or common carriers for feeds, binders, anti-caking agents, stabilizers, emulsifiers, diluents, solvents or combinations thereof.

The "carrier" of the present invention refers to a feedable material capable of carrying an active ingredient, improving its dispersibility, and having good chemical stability and adsorptivity, and is divided into an organic carrier and an inorganic carrier. The organic carrier is a material containing much crude fiber, and includes but is not limited to corn flour, corn cob powder, wheat bran, rice hull powder, defatted rice bran, unite bran, corn straw powder, peanut hull powder and the like. The inorganic carrier is generally mineral substance, mainly divided into calcium salt and silicon oxide, and is used for preparing microelement premix including but not limited to calcium carbonate, silicate, vermiculite, zeolite, sepiolite and the like.

The diluent is a substance which uniformly distributes additive raw materials in a material, dilutes the additive raw materials with high concentration into premix or premix with low concentration, can separate trace components from each other, and reduces the mutual reaction among active components so as to increase the stability of the active components without influencing the physicochemical properties of the related substances. The diluent is classified into organic diluents and inorganic diluents, and common organic diluents include, but are not limited to, corn flour, degermed corn flour, dextrose (glucose), sucrose, bran-bearing raw wheat flour, parched soybean flour, wheat middling, corn gluten meal, etc., and common inorganic diluents include, but are not limited to, limestone, monocalcium phosphate, shell powder, kaolin (kaolin), salt and sodium sulfate.

Adjuvants contemplated by the present invention include, but are not limited to, binders, wetting agents, disintegrants, lubricants, antioxidants, preservatives.

The term "vehicle" as used herein refers to the solvent required to dissolve or disperse the solid, including but not limited to water, ethanol, glycerol, and the like.

Further, the feed composition comprises an additional animal feed additive selected from the group consisting of a nutritional feed additive, a general feed additive, and a pharmaceutical feed additive.

Specifically, the nutritional feed additives include, but are not limited to, amino acids, amino acid salts and analogs thereof, vitamins and retinoids, mineral elements and complexes thereof, microbial enzyme preparations or nonprotein nitrogen; the general feed additives include, but are not limited to, growth promoters, insect repellent health agents, flavoring and feeding promoting agents, feed conditioners, feed storage agents and herbal additives; the medicinal feed additive includes, but is not limited to, veterinary drug premix substances which have the effects of preventing animal diseases and promoting animal growth and can be added into feed for long-term use and mixed with carriers or diluents.

Furthermore, the feed composition can comprise feed raw materials, and the feed raw materials are selected from feed substances, such as animals, plants, microorganisms or minerals, which are not feed additives and can be used for processing and preparing feed.

In some embodiments, the feed composition is an additive premix, concentrate, compound feed, or concentrate supplement.

The feed additive premix feed is a uniform mixture which is prepared by taking two or more nutritional feed additives of mineral trace elements, vitamins, microorganisms and amino acids as main materials and butyrylglutamic acid derivatives or other feed additives, carriers and/or diluents provided by the invention according to a certain proportion, wherein the content of the nutritional feed additives can meet the basic nutritional requirements of the animals at a specific physiological stage, and the additive amount in compound feed, concentrate supplement or animal drinking water is not less than 0.1% and not more than 10%.

The animal feed may be prepared according to methods well known in the art for different animals and may for example comprise one or more of the following ingredients: corn, sorghum, wheat, barley, soy, pea, fish, dairy, fats and oils, vitamins and minerals, and the like.

The concentrated feed is prepared by mainly mixing protein, mineral substances and feed additives according to a certain proportion.

The compound feed is prepared by mixing a plurality of feed raw materials and feed additives according to a certain proportion according to the nutritional requirements of bred animals.

The concentrated feed supplement is a feed prepared from various feed raw materials and feed additives according to a certain proportion for supplementing the nutrition of the herbivore.

The invention also relates to a preparation process of the feed composition, which comprises the steps of weighing raw materials and auxiliary materials, mixing by a mixing unit, granulating, inspecting quality and packaging.

Drawings

Fig. 1 is a flow diagram of a process for preparing a mixed pellet feed additive, indicating key control points.

The specific implementation mode is as follows:

in order to make the objects, technical solutions and advantages of the present invention more apparent, the compounds, compositions and applications of the present invention are further described in detail by examples below. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Firstly, preparing a butyrylglutamic acid derivative:

example 1 Butyrylglutamic acid

200.34g (1.36mol,1eq) of L-glutamic acid and 850mL (136.00g,3.40mol,2.5eq) of 4M NaOH aqueous solution are sequentially added into a 2L three-neck flask, stirred at room temperature for 0.5 hour, added with 215.44g (1.36mol,1eq.) of butyric anhydride dropwise, added with 4M NaOH aqueous solution dropwise to adjust the pH to 8-9, and monitored by TLC until the reaction is finished. And dropwise adding concentrated hydrochloric acid into the reaction solution to adjust the pH to 2-3, extracting with ethyl acetate (1L multiplied by 3), and combining organic phases. Washing the organic phase with saturated brine (1L multiplied by 3), drying with anhydrous sodium sulfate, concentrating under reduced pressure to obtain a crude product which is a colorless transparent liquid, cooling to 0 ℃ for crystallization, performing suction filtration, washing filter residues with n-heptane (100mL multiplied by 3), and performing vacuum drying at 40 ℃ overnight to obtain a product (butyrylglutamic acid) which is a white solid, wherein the yield is 237.97g and is 79.86%. LC-MS (ESI, neg. ion) M/z:216[ M-H]^-；¹HNMR(400MHz,d₆-DMSO):δ(ppm)0.85(t,3H),1.51(m,2H),1.76(m,1H),1.95(m,1H),2.09(t,2H),2.26(t,2H),4.19(td,1H),8.02(d,1H),12.34(s,2H)。

Example 2N-butyryl-L-glutamic acid diethyl ester

Step 1: synthesis of L-glutamic acid diethyl ester hydrochloride

To a 1000mL single-neck flask was added 320mL (5.48mol, 40eq) of absolute ethanol, cooled to-10 ℃ and 28mL (378.92mmol, 2.8eq) of thionyl chloride was slowly added dropwise to the reaction system. After the completion of the dropwise addition, the reaction mixture was stirred at a constant temperature of-10 ℃ for 1 hour, and then 20.08g (136.48mmol, 1eq) of L-glutamic acid was added to the reaction system. The reaction was stirred at room temperature for 2 hours, then warmed to 80 ℃ for reaction, and monitored by TLC to the end. The reaction solution was concentrated under reduced pressure to remove the solvent, cooled in a refrigerator for crystallization, washed with ethyl acetate (150 mL. times.3), separated by suction filtration, and dried to give the product (L-glutamic acid diethyl ester hydrochloride) as a white solid (25.44 g), with a yield of 77.76%. LC-MS (ESI, pos.ion) M/z 204[ M-Cl]⁺；¹HNMR(400MHz,D₂O):δ(ppm)1.20(t,3H),1.25(t,3H),2.19(m,2H),2.58(m,2H),4.12(m,3H),4.24(q,2H).

Step 2: synthesis of N-butyryl-L-glutamic acid diethyl ester

25.44g (106.13mmol,1eq) of L-glutamic acid diethyl ester hydrochloride is weighed into a 500mL single-neck flask, added with 150mL of dichloromethane for dissolution, added with 26.93g (266.13mmol,2.5eq) of triethylamine, cooled to 0 ℃, then dissolved with 17.07g (160.21mmol,1.5eq) of butyryl chloride in 50mL of dichloromethane, slowly added dropwise to make the temperature of the reaction system about 0 ℃, stirred at 0 ℃ for 1 hour after the dropwise addition is finished, and then reacted at room temperature for 2 hours to stop the reaction. The reaction system was quenched by adding 100mL of water, the organic phase was separated, the aqueous phase was extracted with methylene chloride (100 mL. times.2), the organic phases were combined, the organic phase was washed successively with water (300 mL. times.3) and saturated brine (300 mL. times.3), dried over anhydrous sodium sulfate for 30 minutes, and concentrated under reduced pressure to give N-butyryl-L-glutamic acid diethyl ester as a colorless liquid, a yield of 16.00g, and a yield of 55.36%. LC-MS (ESI, pos.ion) M/z 274.2[ M + H ]]⁺；¹HNMR(400MHz,CDCl₃):δ(ppm)0.97(t,3H),1.26(t,3H),1.30(t,3H),1.68(m,2H),2.00(m,2H),2.21(m,3H),2.40(m,2H),4.14(q,2H),4.21(q,2H),4.62(td,1H),6.19(d,1H)。

Example 3N-butyryl-L-glutamic acid di-N-butyl ester

Step 1: synthesis of L-glutamic acid di-n-butyl ester hydrochloride

403.03g (5.44mol, 40eq) of n-butanol was added to a 1000mL single-neck flask, cooled to-10 ℃ and 45.28g (380.62mmol, 2.8eq) of thionyl chloride was slowly added dropwise to the reaction system. After the completion of the dropwise addition, the reaction mixture was stirred at a constant temperature of-10 ℃ for 1 hour, and then 20.00g (135.93mmol, 1eq) of L-glutamic acid was added to the reaction system. The reaction was stirred at room temperature for 2 hours, then warmed to 85 ℃ for reaction, and monitored by TLC to the end. The reaction solution was concentrated under reduced pressure to remove the solvent, cooled in a refrigerator to crystallize, washed with ethyl acetate (150 mL. times.3), separated by suction filtration, and dried to give the product (L-glutamic acid di-n-butyl ester hydrochloride) as a white solid (22.06 g), with a yield of 62.33%. LC-MS (ESI, pos.ion) M/z 260.1[ M-Cl ]]⁺。

Step 2: synthesis of N-butyryl-L-glutamic acid di-N-butyl ester

22.00g (74.37mmol,1eq) of L-glutamic acid di-n-butyl ester hydrochloride was weighed in a 500mL single-neck flask, dissolved by adding 150mL of dichloromethane, added with 18.81g (185.93mmol,2.5eq) of triethylamine, cooled to 0 ℃, then 11.89g (111.56mmol,1.5eq) of butyryl chloride was dissolved in 50mL of dichloromethane, slowly added dropwise so that the temperature of the reaction system was around 0 ℃, after the addition was completed, stirred at 0 ℃ for 1 hour, then reacted at room temperature for 2 hours, and the reaction was stopped. The reaction system was quenched by adding 100mL of water, the organic phase was separated, the aqueous phase was extracted with dichloromethane (100 mL. times.2), the organic phases were combined, the organic phase was washed successively with water (300 mL. times.3) and saturated brine (300 mL. times.3), dried over anhydrous sodium sulfate for 30 minutes, and concentrated under reduced pressure to give N-butyryl-L-glutamic acid di-N-butyl ester as a colorless liquid, yield 14.11g, yield 57.58%. LC-MS (ESI, pos.ion) M/z 330.3[ M + H ]]⁺。

Example 4N-butyryl-D-glutamic acid di-N-butyl ester

The procedure was as in example 2.

The glutamic acid used in step 1 was D-glutamic acid, and the reaction product was a white solid, with a yield of 20.27g and a yield of 57.28%. LC-MS (ESI, pos.ion) M/z 260.0[ M-Cl ]]⁺。

The reaction product of step 2, N-butyryl-D-glutamic acid di-N-butyl ester, was a colorless liquid, yield 14.48g, 59.11%. LC-MS (ESI, pos.ion) M/z 330.4[ M + H ]]⁺。

Example 5N-butyryl-L-glutamic acid di-N-hexyl ester

Step 1: synthesis of L-glutamic acid di-n-hexyl ester hydrochloride

41.67g (407.8mmol, 3.0eq) of hexanol and 400mL of toluene are added into a 1000mL single-neck bottle in sequence, cooled to minus 10 ℃ and reversely reactedTo the reaction system, 45.28g (380.62mmol, 2.8eq) of thionyl chloride was slowly added dropwise. After the completion of the dropwise addition, the reaction mixture was stirred at a constant temperature of-10 ℃ for 1.5 hours, and then 20.00g (135.93mmol, 1eq) of L-glutamic acid was added to the reaction system. The reaction was stirred at room temperature for 2 hours, then warmed to 85 ℃ for reaction, and monitored by TLC to the end. The reaction solution was concentrated under reduced pressure to remove the solvent, cooled in a refrigerator for crystallization, washed with ethyl acetate (150mL × 3), filtered with suction, and dried to give 32.88g of a white solid (L-glutamic acid di-n-hexyl ester hydrochloride) with a yield of 68.74%. LC-MS (ESI, pos.ion) M/z 316.4[ M-Cl]⁺。

Step 2: synthesis of N-butyryl-L-glutamic acid di-N-hexyl ester

22.00g (62.52mmol,1.0eq) of L-glutamic acid di-n-hexyl ester hydrochloride is weighed into a 1000mL single-mouth bottle, 200mL of dichloromethane is added for dissolution, 15.82g (156.29mmol,2.5eq) of triethylamine is added for dissolution, the solution is cooled to 0 ℃, then 9.99g (93.77mmol,1.5eq) of butyryl chloride is dissolved in 50mL of dichloromethane, the solution is slowly dripped to make the temperature of the reaction system about 0 ℃, after the dripping is finished, the solution is stirred at 0 ℃ for 1 hour, then the reaction is carried out at room temperature, and TLC monitors the reaction until the reaction end point. 100mL of water was added to the reaction system to quench the reaction, the organic phase was separated, the aqueous phase was extracted with dichloromethane (100 mL. times.2), the organic phases were combined, the organic phase was washed successively with water (400 mL. times.3) and saturated brine (400 mL. times.3), dried over anhydrous sodium sulfate for 30 minutes, and concentrated under reduced pressure to give N-butyryl-L-glutamic acid di-N-hexyl ester as a colorless liquid, yield 14.98g, yield 62.17%. LC-MS (ESI, pos.ion) M/z 386.6[ M + H [ ]]⁺。

Example 6 bis-N-octyl N-butyryl-L-glutamate

The preparation method is the same as example 4.

The raw materials sequentially charged in the step 1 are 53.11g of n-octanol (407.8mmol, 3.0eq), 420mL of toluene, 45.28g of thionyl chloride (380.62mmol, 2.8eq) and 20.00g of L-glutamic acid (135.93mmol, 1eq), and the product L-glutamic acid di-n-octyl ester hydrochloride is gray solid, the yield is 38.76g and the yield is 69.89%. LC-MS (ESI, pos.ion) m/z:372.5[M-Cl]⁺。

The raw materials sequentially fed in the step 2 are 22.00g (53.92mmol,1.0eq) of L-glutamic acid di-N-octyl ester hydrochloride, 300mL of dichloromethane, 13.64g (134.80mmol,2.5eq) of triethylamine and 8.62g (80.88mmol,1.5eq) of butyryl chloride, and the product N-butyryl-L-glutamic acid di-N-octyl ester is light yellow oily matter, the yield is 16.96g, and the yield is 71.20%. LC-MS (ESI, pos.ion) M/z 442.4[ M-H ]]⁺。

Example 7N-butyryl-L-glutamic acid dilaurate

The preparation method is the same as example 4.

The raw materials sequentially put into the step 1 are 53.19g (285.46mmol, 2.1eq) of lauryl alcohol, 420mL of tetrahydrofuran, 45.28g (380.62mmol, 2.8eq) of thionyl chloride and 20.00g (135.93mmol, 1eq) of L-glutamic acid, and the product of the L-glutamic acid dilaurate hydrochloride is a white solid, the yield is 41.48g, and the yield is 58.65%. LC-MS (ESI, pos.ion) M/z 484.9[ M-Cl]⁺。

The raw materials sequentially charged in step 2 were L-glutamic acid dilaurate hydrochloride 22.00g (42.29mmol,1.0eq), 300mL of dichloromethane, triethylamine 10.70g (105.72mmol,2.5eq), and 6.76g (63.43mmol,1.5eq) of butyryl chloride, and the product N-butyryl-L-glutamic acid dilaurate was a yellow oil with a yield of 12.80g and a yield of 54.67%. LC-MS (ESI, pos.ion) M/z 554.9[ M-H]⁺。

Example 8N-butyryl-L-glutamic acid dipalmityl ester

The preparation method is the same as example 4.

The raw materials sequentially charged in the step 1 are 79.75g of palmitol (328.96mmol, 2.1eq), 450mL of tetrahydrofuran, 45.28g of thionyl chloride (380.62mmol, 2.8eq) and 20.00g of L-glutamic acid (135.93mmol, 1.0eq), and the product L-glutamic acid dipalmitate hydrochloride is a light yellow solid, the yield is 60.06g and 63.51%. LC-MS (ESI, pos.ion) m/z of 583.1[ 2 ]M-Cl]⁺。

The raw materials sequentially charged in step 2 are 22.00g (34.79mmol,1.0eq) of L-glutamic acid dipalmityl ester hydrochloride, 300mL of dichloromethane, 8.80g (86.96mmol,2.5eq) of triethylamine and 5.56g (52.18mmol,1.5eq) of butyryl chloride, the product is a yellow solid, the yield of N-butyryl-L-glutamic acid dipalmityl ester is 11.27g, and the yield is 48.63%. LC-MS (ESI, pos.ion) M/z 667.1[ M-H]⁺。

Example 9N-butyryl-L-glutamic acid monoethyl ester

Step 1: preparation of L-glutamic acid monoethyl ester

50.10g (340.52mmol,1.0eq) of L-glutamic acid is dissolved in 473.4g (10.28mol,30.2eq) of ethanol, cooled to 0 ℃, 46.00g (469.01mmol,1.4eq) of concentrated sulfuric acid is added dropwise, after the dropwise addition is finished, the mixture is naturally cooled to room temperature for reaction overnight, and the TLC monitors the reaction to the end point. Cooling the reaction liquid to 0-5 ℃, dropwise adding triethylamine to the pH value of 7-8, continuously stirring at room temperature for 2 hours after dropwise adding, performing suction filtration, recrystallizing a filter cake with an ethanol water solution (ethanol/water (v: v) ═ 4:1, 250mL), performing suction filtration, and performing vacuum drying on filter residues at 40 ℃ overnight to obtain a product, namely the ethyl L-glutamate, which is a white solid, wherein the yield is 43.00g and 72.08%. LC-MS (ESI, pos.ion) M/z 176[ M + H ]]⁺；¹HNMR(400MHz,CDCl₃):δ(ppm)1.16(t,3H),2.07(m,2H),2.47(m,2H)),3.68(t,1H),4.14(q,2H),4.08(q,2H)。

Step 2: preparation of N-butyryl-L-glutamic acid monoethyl ester

2.01g (11.47mmol,1.0eq) of L-glutamic acid monoethyl ester was added to 10mL of glacial acetic acid, and stirred to dissolve, 2.20g (13.91mmol,1.2eq) of butyric anhydride was further added, and the reaction was stirred at room temperature overnight, and monitored by TLC until the end point. The reaction mixture was concentrated under reduced pressure to remove the reaction solvent to give a pale yellow oil, and the product, ethyl N-butyryl-L-glutamate, was subjected to reverse silica gel column chromatography (methanol/water (v: v) ═ 4:1) to give a colorless oil, and the yield was 1.98g and 70.36%. LC-MS (ESI, neg. ion) M/z 244.4[ M-H [ ]]^-。

Example 10N-butyryl-L-glutamic acid monobutyl ester

The preparation method is the same as example 8.

The raw materials sequentially fed in the step 1 are 50.10g (340.52mmol,1.0eq) of L-glutamic acid, 640.0g (8.51mol,25.0eq) of n-butanol and 46.00g (469.01mmol,1.4eq) of concentrated sulfuric acid, the product L-mono butyl glutamate is white solid, the yield is 60.0g, and the yield is 73.10%. LC-MS (ESI, pos.ion) M/z 204.1[ M + H ]]⁺。

The starting materials sequentially charged in step 2 were 20mL of glacial acetic acid, 5.01g (24.60mmol,1.0eq) of L-glutamic acid monobutyl ester and 4.67g (29.52mmol,1.2eq) of butyric anhydride, and the product N-butyryl-L-glutamic acid monobutyl ester was a colorless oil by reverse silica gel column chromatography during the work-up (methanol/water (v: v) ═ 9:2), with a yield of 4.49g and a yield of 66.77%. LC-MS (ESI, neg. ion) M/z 272.2[ M-H [)]^-。

Example 11 Monooctyl N-butyryl-L-glutamate

The preparation method is the same as example 8.

The raw materials sequentially fed in the step 1 comprise 300mL of N, N-dimethylformamide, 50.10g (340.52mmol,1.0eq) of L-glutamic acid, 53.21g (408.62mmol,1.2eq) of N-octanol and 46.80g (476.72mmol,1.4eq) of concentrated sulfuric acid, and the product L-monooctyl glutamate is a white solid, the yield is 61.41g, and the yield is 69.54%. LC-MS (ESI, pos.ion) M/z 260.3[ M + H ]]⁺。

The raw materials sequentially charged in step 2 were 20mL of glacial acetic acid, 5.01g (19.28mmol,1.0eq) of monooctyl L-glutamate and 3.66g (23.14mmol,1.2eq) of butyric anhydride, and the product N-butyryl-monooctyl-glutamate was a white solid by reverse silica gel column chromatography (methanol/water (v: v) ═ 5:1) in the post-treatment, with a yield of 3.54g and a yield of 55.70%. LC-MS (ESI, neg. ion) M/z 328.2[ M-H ]]^-。

Example 12N-butyryl-L-glutamic acid monolauryl ester

The preparation method is the same as example 8.

The raw materials sequentially fed in the step 1 comprise 300mL of N, N-dimethylformamide, 50.00g (339.84mmol,1.0eq) of L-glutamic acid, 75.99g (407.80mmol,1.2eq) of lauryl alcohol and 46.7g (475.8mmol,1.4eq) of concentrated sulfuric acid, and the product L-glutamic acid monolaurate is light yellow solid, the yield is 62.5g, and the yield is 58.3%. LC-MS (ESI, pos.ion) M/z 316.2[ M + H ]]⁺。

The raw materials sequentially added in the step 2 are 20mL of glacial acetic acid, 5.00g (15.85mmol,1.0eq) of L-monolauryl glutamate and 3.01g (19.02mmol,1.2eq) of butyric anhydride, and the product N-butyryl-L-monolauryl glutamate is white solid by reverse silica gel column chromatography (methanol/water (v: v) ═ 5:1) in the post-treatment, the yield is 4.12g, and the yield is 67.40%. LC-MS (ESI, neg. ion) M/z 384.3[ M-H [)]^-。

Example 13N-butyryl-L-glutamic acid monopalmityl ester

The preparation method is the same as example 8.

The raw materials sequentially charged in the step 1 are 300mL of N, N-dimethylformamide, 50.00g (339.84mmol,1.0eq) of L-glutamic acid, 98.87g (407.80mmol,1.2eq) of palmityl alcohol and 46.7g (475.8mmol,1.4eq) of concentrated sulfuric acid, and the product L-monopalmityl glutamate is a light yellow solid, the yield is 76.0g, and the yield is 60.2%. LC-MS (ESI, pos.ion) M/z 372.3[ M + H [ ]]⁺。

The raw materials sequentially charged in step 2 were glacial acetic acid 30mL, L-glutamic acid monopalmitate 5.00g (13.46mmol,1.0eq) and butyric anhydride 2.55g (16.15mmol,1.2eq), and the product of reverse silica gel column chromatography (methanol/water (v: v) ═ 6:1) in the post-treatment was a white solid, the yield of N-butyryl-L-glutamic acid monopalmitate was 3.1g, and the yield was 52.1%. LC-MS (ESI, neg. ion) M/z 440.4[ M-H [)]^-。

The preparation method of the feed composition comprises the following steps:

the butyrylglutamic acid derivatives of examples 1 to 13 were mixed with corresponding adjuvants to prepare corresponding mixed type pellet feed additives.

1. Material

Raw materials: any one of the butyrylglutamic acid derivatives, butyrylglutamic acid, butyric acid and glutamic acid of examples 1 to 13;

carrier: corn starch;

adhesive: 1.3% hydroxypropyl methylcellulose in water.

2. Product formula

TABLE 1 formulation of mixed type pellet feed additive of butyrylglutamic acid derivatives

3. Production process

The production process of the mixed type pellet feed additive according to the present invention will be described with reference to fig. 1.

Raw material supply: the product is qualified and has a purity of more than or equal to 99% according to the inspection of the product control department by research and development center of Guangzhou Ensaite Biotechnology Limited.

Auxiliary material purchase: auxiliary materials are purchased from qualified suppliers, sampled after the auxiliary materials are delivered to a factory, and can be warehoused and stored for later use after being checked to be qualified, and the process is a key control point and the quality of the auxiliary materials must be strictly controlled.

Material production and weighing: according to the proportion of the product formula, the raw materials and the auxiliary material carriers are sequentially weighed and rechecked, and the generated wastes (packaging bags) are uniformly stored and treated.

Mixing: the raw materials and the auxiliary materials are put into a mixing unit to be uniformly mixed, the step is a key control point, the mixing time must be strictly controlled, and the mixing uniformity is regularly verified. The mixer is equipped with a pulse dust collector to reduce dust.

And (3) granulating: putting the product obtained by mixing the raw materials and the auxiliary materials and a 1.3% hydroxypropyl methyl cellulose aqueous solution into a granulator according to the mass ratio of 100:35, starting a mixing and cutting knife to operate for 3-5min, drying the granulated material in a fluidized bed, and sieving the granulated material with a 16-mesh sieve after 30 min.

Packaging and inspecting: weighing and packaging according to the packaging specification, storing in a finished product bin, identifying information such as production batches and inspection states, and at least sampling and inspecting two samples in each production batch to a laboratory for delivery inspection and sample reservation. Can leave the factory after being qualified. Thus obtaining the mixed type pellet feed additive.

Cleaning production equipment: after each batch of products is produced, the production area must be cleaned. When the production varieties are replaced, the carrier operation and impurity removal must be carried out on the production equipment, so that the cross contamination is prevented.

II, animal breeding test:

example A Effect of different feed additives in broiler feed

The test adopts single-factor random design, 480 three yellow-feathered broilers with age of 22 days, average weight of 145g and similar weight are selected and randomly divided into 4 treatment groups, each group has 6 repetitions, each group has half of a male parent, and each repetition has 20 yellow-feathered broilers. The chicken coop and the appliances were sterilized before the test. In the test period, cage culture is carried out in the same chicken house under the same feeding management condition. The basic daily ration mainly comprises corn-soybean meal, and other antioxidant ingredients and growth promoters are not additionally added in the whole feeding process. Each test group is a control group and 1-3 test groups. Wherein the control group only gives basic ration, and the test 1-3 groups respectively add 2000ppm of the mixed type pellet feed additive control 1 (glutamic acid), the control 2 (butyric acid) and the product 1 (butyrylglutamic acid) in the basic ration. The test period is 20 days, and the test chicken freely drinks and takes food and feeds for 2 times a day. Weighing each repetition at 42 days of age (stopping feed for 12h without stopping water), counting the feed consumption of the test chickens, and calculating the Average Daily Feed Intake (ADFI), Average Daily Gain (ADG) and Feed Conversion Ratio (FCR) of each group of test chickens. Statistical analysis is carried out on test data by adopting SPSS18 software, single-factor analysis of variance (ANOVA) is carried out on the data, if the difference between treatments is obvious, multiple comparison is carried out by using a Duncan's method, and the significance level is 0.05. The test results are expressed as "mean. + -. standard error" and are shown in Table 3. From the results, the average daily feed intake and the average daily gain of the test chickens in the test 1 group and the test 2 group are improved to different degrees, but the feed-meat ratio is not obviously reduced; the average daily feed intake of the test chickens in the test group 3 is similar to that of the control group, the average daily gain is improved, and the feed-meat ratio is reduced by 5.26%.

TABLE 3 influence of different feed additives on broiler productivity results

Test article

Number of trials

ADFI(g)

ADG(g)

FCR

Control group

-

20*6

623.00±9.24

205.00±4.80

3.04±0.05

Test 1 group

Reference

1

20*6

678.50±9.14

226.33±2.86

3.00±0.03

Test 2 groups

Control

2

20*6

663.83±10.60

220.00±3.18

3.02±0.01

Test 3 groups

Product

1

20*6

623.50±13.63

216.33±2.24

2.88±0.02

Example B application Effect of butyrylglutamic acid derivatives in broiler feed

The test adopts single-factor random design, 1680 three yellow-feathered broilers with age of 22 days, average weight of 153g and similar weight are selected and randomly divided into 14 treatment groups, each group has 6 repetitions, each half of the male and female groups has 20 repetitions. The chicken coop and the appliances were sterilized before the test. In the test period, cage culture is carried out in the same chicken house under the same feeding management condition. The basic daily ration mainly comprises corn-soybean meal, and other antioxidant ingredients and growth promoters are not additionally added in the whole feeding process. Each test group is a control group and 1-13 test groups. Wherein the control group only gives basic ration, and the test 1-13 groups respectively add 2000ppm of the mixed type pellet feed additive product 1-13 of the butyrylglutamic acid derivative provided by the invention in the basic ration. The test period is 20 days, and the test chicken freely drinks and takes food and feeds for 2 times a day. Counting the diarrhea rate of the test chicken by taking each repetition as a unit; weighing at 42 days of age (stopping feed for 12h without stopping water), counting the feed consumption of test chickens, and calculating the Average Daily Feed Intake (ADFI), Average Daily Gain (ADG) and Feed Conversion Ratio (FCR) of each group of test chickens. Statistical analysis is carried out on test data by adopting SPSS18 software, single-factor analysis of variance (ANOVA) is carried out on the data, if the difference between treatments is obvious, multiple comparison is carried out by using a Duncan's method, and the significance level is 0.05. The test results are expressed as "mean. + -. standard error" and are shown in Table 4. From the results, the butyrylglutamic acid product can improve the feed intake of the experimental chicken but is not significant compared with a blank control group, and the butyrylglutamic acid derivative has no influence on the average daily feed intake of the experimental chicken; the butyrylglutamic acid and the derivative thereof can obviously improve the average daily gain of the test chicken and reduce the feed conversion ratio by 4.6-13.7% on the premise of not influencing the feed intake of the test chicken, and simultaneously the diarrhea rate of the test chicken is effectively controlled.

TABLE 4 influence of butyrylglutamic acid derivatives on broiler productivity

Example C application study of butyrylglutamic acid derivatives in pig feed

390 big and large triple lean piglets with similar body weight of 65 days old are randomly divided into 13 treatment groups, each group is 3 in repetition, each half of male and female is 10 in each group. Pigsty and utensils were sterilized before the test. In the test period, the pigs are housed in different columns under the same feeding and management conditions in the same pigpen. During the test period, test pigs were fed free food and water 2 times a day. Each test group is a control group and a test group 1-12. Wherein, the control group only gives the basic ration, and the test 1-12 groups respectively give the mixed type granular feed additive products 1-13 which respectively add 2500ppm of the butyrylglutamic acid derivative provided by the invention in the basic ration. In the whole feeding process, other antioxidant ingredients and growth promoters are not additionally added in each test group. The test period is 28 days, and the production performance of the test pigs is counted by taking each repetition as a unit, namely Average Daily Feed Intake (ADFI), Average Daily Gain (ADG), Feed Conversion Ratio (FCR) and diarrhea rate. The test results are shown in table 5. Statistical analysis is carried out on test data by adopting SPSS18 software, single-factor analysis of variance (ANOVA) is carried out on the data, if the difference between treatments is obvious, multiple comparison is carried out by using a Duncan's method, and the significance level is 0.05. The test results are expressed as "mean. + -. standard error" and are shown in Table 5. From the results, it is understood that the feed intake of the test pigs was not significantly affected in the test group to which the butyrylglutamic acid derivative was administered, but the average daily gain was significantly increased, the feed-meat ratio was decreased by 7.6% to 12.5%, and the diarrhea rate was decreased by 10.0% to 30.9%.

TABLE 5 influence of butyrylglutamic acid derivatives on pig Productivity

Test article

Number of trials

ADFI(kg)

ADG(kg)

FCR

Rate of diarrhea/%)

Control group

-

10*3

11.87±0.32

4.51±0.08

2.63±0.02

32.33±1.45

Test 1 group

Product 2

10*3

12.73±0.33

5.23±0.06

2.43±0.03

27.00±0.58

Test 2 groups

Product 3

10*3

12.40±0.26

5.23±0.12

2.37±0.01

26.00±0.58

Test 3 groups

Product 4

10*3

12.77±0.20

5.42±0.09

2.36±0.01

25.00±0.58

Test 4 groups

Product 5

10*3

12.53±0.32

5.42±0.16

2.31±0.01

23.00±0.58

Test 5 groups

Product 6

10*3

12.53±0.27

5.45±0.15

2.30±0.02

22.00±0.58

Test 6 groups

Product 7

10*3

12.43±0.43

5.40±0.21

2.30±0.01

19.33±0.33

Test 7 groups

Product 8

10*3

12.500.38

5.40±0.19

2.31±0.01

19.00±0.58

Test 8 groups

Product 9

10*3

12.70±0.26

5.16±0.13

2.46±0.02

27.00±0.58

Test 9 groups

Product 10

10*3

12.60±0.66

5.19±0.25

2.43±0.01

26.00±0.58

Test 10 groups

Product 11

10*3

12.33±0.34

5.22±0.12

2.42±0.01

24.00±0.58

Test 11 groups

Product 12

10*3

12.27±0.22

5.24±0.09

2.36±0.02

23.00±0.58

Test 12 groups

Product 13

10*3

12.47±0.61

5.33±0.26

2.34±0.01

22.00±0.58

Example D application Effect of Butyrylglutamic acid derivative in meat Duck feed

The experiment adopts single-factor random design, 1560 cherry valley meat ducks with the age of 20 days, the average weight of 1300g and the similar weight are selected and randomly divided into 13 treatment groups, each group has 6 repetitions, each half of the male and female ducks has 20 repetitions. The duck shed and the apparatus were sterilized before the test. In the experimental period, cage culture is carried out under the same feeding and management conditions in the same duck shed. The basic daily ration mainly comprises corn-cottonseed meal, and other antioxidant ingredients and growth promoters are not additionally added in the whole feeding process. Each test group is a control group and 1-12 test groups. Wherein the control group only gives basic ration, and the test 1-12 groups respectively add 2000ppm of the mixed type pellet feed additive product of the butyrylglutamic acid derivative provided by the invention in the basic ration, wherein the mixed type pellet feed additive product is 2-13. The test period is 28 days, the test ducks drink water and feed freely, and the feed is carried out for 2 times a day. Weighing each repetition as a unit at the age of 48 days (stopping feed for 12h without stopping water), counting the feed consumption of the test ducks, and calculating the Average Daily Feed Intake (ADFI), Average Daily Gain (ADG) and Feed Conversion Ratio (FCR) of each group of test ducks. Statistical analysis is carried out on test data by adopting SPSS18 software, single-factor analysis of variance (ANOVA) is carried out on the data, if the difference between treatments is obvious, multiple comparison is carried out by using a Duncan's method, and the significance level is 0.05. The test results are expressed as "mean. + -. standard error" and are shown in Table 6. From the results, the change of the production performance of the meat duck living under the same feeding condition by the butyrylglutamic acid derivative is very obvious compared with the control group, the change is shown in the aspects of obvious improvement of average daily gain and obvious reduction of feed conversion ratio, and the conversion rate of the feed can be obviously improved by adding the butyrylglutamic acid derivative into the basic ration of the meat duck.

TABLE 6 influence of butyrylglutamic acid derivatives on meat duck productivity

Example E use of butyrylglutamic acid derivatives in aquatic feed

(1) Test materials

The fish used for the test: the test fish is grass carp, the current year fingerling, and a Dafeng fingerling farm in Huizhou, Guangdong province. Healthy and active grass carp seeds with consistent specificationIn a large net cage (4X 2X 1.5 m)³) The culture is carried out for 4 weeks before the culture is carried out for formal culture tests, and the experimental system is a small floating net cage (the specification is 1.1 multiplied by 1.1 m)³) Each small net cage is provided with an inflation head, and 24 hours of air inflation is carried out every day. The small net cage and the temporary culture net cage are placed in a test field with a height of 3500m²The depth of the pond water is 1.5m, and the pond water is fully aerated bottom water. During the test, the black carp 624 hungry for 1d is randomly divided into 13 groups, each group is provided with 4 repeats, 12 fishes are repeatedly placed in each repeat, the whole fish is weighed and then randomly placed in 52 net cages to feed different test feeds respectively.

Test feed: the experimental feeds were prepared according to the formulation of table 7, and different test groups were added with the same amount of butyrylglutamic acid derivatives according to table 8. The feed raw materials are subjected to superfine grinding and then are prepared into buoyancy expanded feed with the particle size of 3mm by a Jiangsu shepherd expansion machine set, the mold stripping temperature is 130 ℃, 3% of soybean oil is sprayed outside by oil spraying equipment, and the feed is sealed and stored in a shade place for later use.

Table 7 grass carp feed formulation and chemical composition (% wt.) for testing

Composition of raw materials	Content (%)	Composition of raw materials	Content (%)
				Fish meal	9.0	Soybean oil	3.0
Sausage casing powder	3.0	Phosphatide rapeseed meal	9.0
				Bean pulp	12.0	Wheat gluten	4.0
Rapeseed meal	12.0	Blood cell powder	2.0
				Monosodium glutamate protein	3.0	Vc-phosphoric acid ester	0.1
Wheat middling	12.6	Calcium dihydrogen phosphate	1.8
				Flour	17.0	Choline chloride	0.2
Bentonite clay	0.70	Multi-dimensional	0.1
				Rice bran	10.0	Micro-mineral premix	0.5

TABLE 8 growth promotion test grouping of butyrylglutamic acid derivatives

(2) Test method

And (3) test management: the experiment adopts artificial food restriction feeding, the feeding amount is adjusted once per week, the feeding level (according to the initial weight) of each group is completely consistent, and the feeding is carried out twice per day (7:30 and 15: 00). The test period was 8 weeks. The water quality is monitored regularly during the test period, the water temperature is 26.88 +/-3.08 ℃ and DO is kept in the whole cultivation process>5.0mg O L^-1pH 7.8, ammonia nitrogen<0.50mg N L^-1Nitrite nitrogen<0.05mg N L^-1。

And (3) parameter statistics: during the test, after the feeding is stopped for 1d, the net cages are weighed integrally, and the weight gain rate (WG,%), the Feed Coefficient (FCR) and the survival rate (SR,%) are calculated. The calculation formula is as follows:

weight gain (WG,%) 100 × (average end weight-average initial weight)/average initial weight;

feed Factor (FCR) feed intake/fish body weight gain;

survival (SR,%) 100 x number of fish at the end of the test/number of fish at the beginning of the test.

(3) Test results

The results of the growth promotion test of butyrylglutamic acid derivatives are shown in Table 9. The results show that the test group added with the butyrylglutamic acid derivative is superior to the blank control group without the butyrylglutamic acid derivative in weight gain and feed coefficient, and has obvious growth promoting effect.

TABLE 9 application test results of butyrylglutamic acid derivatives in aquatic feed

The above-described embodiments are merely illustrative of some of the various ways in which the invention may be practiced and are intended to be covered by other embodiments. Accordingly, the embodiments of the present invention will be described by way of illustration, but not construed to limit the scope of the invention, and modifications made within the scope and spirit of the invention or equivalents added to the claims are possible.

Claims

1. A butyrylglutamic acid derivative having a structure represented by the following general formula, or a racemate, a tautomer or a feed-acceptable salt thereof:

the substituent groups A and B in the formula are respectively H or alkyl, and the alkyl is n-propyl, isopropyl or C₅-C₂₀Straight or branched alkyl, C₁-C₈Straight or branched haloalkyl, or C₃-C₇Cycloalkyl, the radicals A and B not both being H.

2. The butyrylglutamic acid derivative according to claim 1, or a racemate, a tautomer or a feed-acceptable salt thereof, wherein the substituent groups a and B are both alkyl groups.

3. The butyrylglutamic acid derivative according to claim 1, or a racemate, a tautomer or a feed-acceptable salt thereof, wherein the substituent groups a and B are not H at the same time.

4. The butyrylglutamic acid derivative of claim 1, or a racemate, a tautomer or a feed-acceptable salt thereof, having one of the following structures:

5. use of the butyrylglutamic acid derivative according to any one of claims 1 to 4, or a racemate, a tautomer or a feed acceptable salt thereof, for the preparation of an animal feed additive.

6. The use of claim 5, wherein the animal comprises poultry, livestock, aquaculture animals or pets.

7. Use of the butyrylglutamic acid derivative according to any one of claims 1 to 4, or a racemate, a tautomer or a feed acceptable salt thereof, for the preparation of an animal feed.

8. A feeding composition comprising the butyrylglutamic acid derivative according to any one of claims 1 to 4, or at least one of racemate, tautomer, or a feed-acceptable salt thereof, and a feedable adjuvant.

9. The feed composition of claim 8, further comprising a feed ingredient.

10. The feed composition as claimed in claim 8 or claim 9 further comprising an additional animal feed additive selected from a nutritional feed additive, a general feed additive or a pharmaceutical feed additive.