A prodrug of a hexahydro-î²-acid compound, feed composition thereof, and use thereof

Abstract

The present invention discloses the prodrugs of hexahydro-β-acid compounds, feed composition thereof, and use thereof, and a prodrug of hexahydro-β-acid compound of formula (I), or solvates thereof, or feed-acceptable salts thereof, wherein, R1 is selected from a substituted or unsubstituted C1-C2 alkyl , and each of R2 and R3 is independently selected from H and a linear or branched C2-C4 carbonyl, wherein the C2-C4 carbonyl is substituted or unsubstituted. Discovered herein is that the the prodrug of a hexahydro-β-acid compound from esterification by aliphatic acid, exhibit stability at high temperature, to overcome the problem resulting from the degradation of hexahydro-β-acid compound in a high-temperature pelleting process. Furthermore, discovered herein is that, both propionate and butyrate, the prodrugs of a hexahydro-β-acid compound from esterification by aliphatic acid, and feed-acceptable salt thereof and solvate thereof, are stable in high-temperature feed processing and achieve effects substantially equal to the hexahydro-β-acid compound in farm breeding.

Description

Precursor compound of hexahydro-β-acid component compound, feed composition and application thereof Technical field:

[0001] The present application relates to the field of animal feed additives, and in particular to a precursor compound of a hexahydro-β-acid component compound and a feed composition containing a precursor compound of the hexahydro-β-acid component compound and applications thereof. Background technology:

[0002] Hop acids are organic acids derived from hops, including α- and β-acids. They have bactericidal, antibacterial, or metabolite-modifying biological effects and are used in animal husbandry as an antibiotic alternative in animal feed. β-acids possess stronger antimicrobial activity. Hop acids have poor stability and solubility. In the past, they were typically ground and added to animal feed, or prepared as a 1% potassium salt solution and sprayed or mixed into the feed. These methods of use are extremely inconvenient in animal husbandry. Studies have suggested that di-, tetra-, or hexa-hydrogenated hop acids could alter their activity, stability, solubility, or other physical properties. Unfortunately, studies have recently reported that hexa-hydrogenated β-acids and their metal salts (hexahydro-β-acids) are heat-labile. Furthermore, when added to feed and stored at room temperature, the hexahydro-β-acids rapidly degrade, resulting in a decrease in their effective content, making them unsuitable for use as feed additives. The inventors discovered in their research that the main components of hexahydro-β-acid are hexahydrolupulone, hexahydrocolupulone, and hexahydro-lupulone. Hexahydro-β-acid is unstable in feed at room temperature because this instability causes a decrease in its content. The inventors further discovered that hexahydro-lupulone or a combination of hexahydro-lupulone and hexahydro-lupulone, when directly used as a mixed feed in animal husbandry, has similar or even superior efficacy in improving animal production performance to hexahydro-β-acid. However, when hexahydro-lupulone and hexahydro-lupulone are used in finished feed, the content of their active ingredients decreases during storage, impacting their use in animal husbandry.

[0003] In view of this, this application is hereby filed.

[0004] Summary of the invention:

[0005] The purpose of the present application includes providing a precursor compound of a hexahydro-β-acid component compound.

[0006] The present application also aims to provide a feed composition comprising a precursor compound of a hexahydro-β-acid component compound.

[0007] The purpose of the present application also includes providing the use of a precursor compound of a hexahydro-β-acid component compound and a feed composition thereof in the preparation of an animal feed additive.

[0008] The purpose of the present application also includes providing a precursor compound of a hexahydro-β-acid component compound and a feed composition thereof for use in preparing animal feed.

[0009] The present application also aims to provide a method for improving animal production performance.

[0010] In order to achieve at least one purpose of this application, the following technical solutions are adopted:

[0011] On the one hand, the present application provides a precursor compound of a hexahydro-β-acid component compound having a structure as shown in formula (I), or a solvate and a feed-acceptable salt thereof, wherein R1 is a substituted or unsubstituted straight-chain or branched alkyl group; R2 or R3 is independently selected from H or a substituted or unsubstituted straight-chain or branched fatty carbonyl group.

[0012]

[0013] On the other hand, the present application also provides a feed composition, which comprises a precursor compound of a hexahydro-β-acid component compound having a structure as shown in formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable excipients.

[0014] On the other hand, the present application also provides a precursor compound of a hexahydro-β-acid component compound having a structure as shown in formula (I), or a solvate thereof and a feed-acceptable salt thereof, and a feed composition comprising a precursor compound of a hexahydro-β-acid component compound having a structure as shown in formula (I), or a solvate thereof and a feed-acceptable salt thereof in the preparation of an animal feed additive.

[0015] On the other hand, the present application also provides a precursor compound of a hexahydro-β-acid component compound having a structure as shown in formula (I), or a solvate thereof and a feed-acceptable salt thereof, and a feed composition comprising a precursor compound of a hexahydro-β-acid component compound having a structure as shown in formula (I), or a solvate thereof and a feed-acceptable salt thereof in the preparation of animal feed.

[0016] On the other hand, the present application also provides a method for improving animal production performance, comprising: feeding the animal with a precursor compound of the hexahydro-β-acid component compound of the present application, or a solvate thereof and a feed-acceptable salt thereof; or, feeding the animal with a feed composition of the present application; or, feeding the animal with a feed containing the feed composition of the present application.

[0017] Compared with the existing technology, the beneficial effects of this application include:

[0018] The present invention has discovered that the fatty acid esterified precursor of the hexahydro-β-acid component compound exhibits excellent stability under high-temperature conditions, overcoming the problem of degradation of the active ingredient of the hexahydro-β-acid component compound during high-temperature pelleting during feed processing. Furthermore, the present invention has discovered that the hexahydro-β-acid component compound propionate or butyrate, and their feed-acceptable salts or solvates, in the fatty acid esterified precursor of the hexahydro-β-acid component compound can withstand the high-temperature process of feed processing and, when used in aquaculture, exhibit substantially equivalent effects to the hexahydro-β-acid component compound.

[0019] Any embodiment of any aspect of the present application may be combined with other embodiments as long as there is no contradiction between them. DETAILED DESCRIPTION

[0020] The above content only summarizes certain aspects of the present application, but is not limited to these aspects. The above content and other aspects will be described in more detail and completely below.

[0021] Further details of this application.

[0022] Certain embodiments of the present application will now be described in detail, examples of which are illustrated by the accompanying structural and chemical formulae. This application is intended to encompass all alternatives, modifications, and equivalent technical solutions, all of which are included within the scope of this application as defined by the claims. Furthermore, certain technical features of the present application are described separately in multiple independent embodiments for clarity, but may also be provided in combination in a single embodiment or in any suitable sub-combination.

[0023] Compound

[0024] The compound involved in the present invention is a precursor compound of a hexahydro-β-acid component compound with a structure shown as formula (I).

[0025]

[0026] wherein R1 is a substituted or unsubstituted straight or branched alkyl group; and R2 and R3 are independently selected from H and a substituted or unsubstituted straight or branched aliphatic carbonyl group.

[0027] Further, R1 is selected from substituted or unsubstituted C1-C2 alkyl; R2 and R3 are independently selected from H and substituted or unsubstituted linear or branched C3-C4 carbonyl.

[0028] The "hexahydro-β-acid component compound" referred to in the present invention refers to the main components contained in hexahydro-β-acid, specifically hexahydrolupulone (I1-1), hexahydrolupulone (I1-2) and hexahydrolupulone (I1-3), and the specific structure is shown in the following formula:

[0029]

[0030] Generally, "substituted" means that one or more substitutable hydrogen atoms in a given structure are replaced by a specified substituent. A substituted group may have a substituent at each substitutable position of the group. When more than one position in a given structure can be substituted by one or more substituents of a specified group, the substituents may be the same or different at each position.

[0031] In the present invention, "C a -C b "Alkyl" means a straight chain or branched saturated alkyl group containing a to b carbon atoms, such as methyl, ethyl, propyl, isopropyl, ..., such as "C3-C4 alkyl" means a straight chain or branched saturated alkyl group containing 3 to 4 carbon atoms. "C a -C b The term "carbonyl" refers to a straight-chain or branched aliphatic carbonyl group containing a to b carbon atoms, such as C(=O)CH2CH3, C(=O)(CH2)2CH3, C(=O)CH2(CH3)2, ...

[0032] In some embodiments, R1 in the precursor compound of the hexahydro-β-acid component compound represented by formula (I) is CH3, and R2 and R3 are independently selected from H and substituted or unsubstituted linear or branched C3-C4 carbonyl.

[0033] Furthermore, R2 and R3 are independently selected from H or unsubstituted straight-chain C3-C4 carbonyl, and are preferably not H at the same time.

[0034] In some embodiments, R3 is H, and R2 is an unsubstituted linear C3-C4 carbonyl group.

[0035] In other embodiments, R2 and R3 are independently selected from unsubstituted straight-chain C3-C4 carbonyl groups, and preferably R2 and R3 are selected from the same unsubstituted straight-chain C3-C4 carbonyl group.

[0036] In some embodiments, R1 in the precursor compound of the hexahydro-β-acid component compound represented by formula (I) is CH2CH3, and R2 and R3 are independently selected from H and substituted or unsubstituted linear or branched C3-C4 carbonyl.

[0037] Furthermore, R2 and R3 are independently selected from H or unsubstituted straight-chain C3-C4 carbonyl, and are preferably not H at the same time.

[0038] In some embodiments, R3 is H, and R2 is an unsubstituted linear C3-C4 carbonyl group.

[0039] In other embodiments, R2 and R3 are independently selected from unsubstituted straight-chain C3-C4 carbonyl groups, and preferably R2 and R3 are selected from the same unsubstituted straight-chain C3-C4 carbonyl group.

[0040] In some specific embodiments, the precursor compound of the hexahydro-β-acid component compound of the present invention includes:

[0041]

[0042]

[0043] Preparation and purification of compounds

[0044] The preparation of the precursor compound of the hexahydro-β-acid component compound represented by formula (I) of the present invention comprises the following process:

[0045] (1) Using commercial hop extract as raw material, β-acid is obtained by organic solvent extraction and separation;

[0046] (2) the β-acid is subjected to hydrogenation reduction to prepare hexahydro-β-acid;

[0047] (3) The hexahydro-β-acid is subjected to recrystallization or chromatographic separation to prepare different hexahydro-β-acid component compounds, wherein the hexahydro-β-acid component compounds are specifically hexahydro-lupulone (I1-1), hexahydro-lupulone (I1-2) and hexahydro-lupulone (I1-3);

[0048] (4) The hexahydro-β-acid component compound is combined with a substituted or unsubstituted straight-chain or branched fatty acid to undergo an esterification reaction to obtain a precursor compound of the hexahydro-β-acid component compound, wherein the esterification reaction includes combining with one chemical equivalent or two chemical equivalents of a substituted or unsubstituted straight-chain or branched fatty acid.

[0049] The hexahydro-β-acid component compound has an asymmetric center and can exist in the form of racemates, racemates, stereoisomers, geometric isomers, tautomers, single enantiomers, single diastereomers, and diastereomers. It should be understood that the precursor compounds of the hexahydro-β-acid component compound obtained by the above step (4) are included in this application. The hexahydro-β-acid component compound can be obtained by chemists with ordinary skills from plant raw materials through semi-synthesis or total synthesis, and can also be obtained through commercial channels.

[0050] In some embodiments, the present invention provides a preparation process of a precursor compound of a hexahydro-β-acid component compound that further involves a separation, purification, or recrystallization process of the reaction product. The reaction product can be obtained as a crude product from the reaction system by a desolvation method. In order to obtain a solid substance with higher chemical purity and lower impurity content, the crude product is dissolved, crystallized, precipitated, or recrystallized and separated in an alcohol solvent, an alcohol-water mixed solvent, or other organic solvent that can be used for product recrystallization under suitable conditions such as temperature, light, and mechanical vibration to obtain a precursor compound of a hexahydro-β-acid component compound with a certain crystalline state. The precursor compound of the hexahydro-β-acid component compound with a certain crystalline state is a precursor compound of the hexahydro-β-acid component compound or a solvate of a precursor compound of the hexahydro-β-acid component compound. The solvate of the precursor compound of the hexahydro-β-acid component compound can be selected from a hydrate of the precursor compound of the hexahydro-β-acid component compound or an ethanolate of the precursor compound of the hexahydro-β-acid component compound.

[0051] As used herein, a "solvate" refers to a eutectic association formed when a compound of the present invention and solvent molecules come into contact, resulting from the binding of stoichiometric or non-stoichiometric amounts of solvent molecules by non-covalent intermolecular forces, caused by external and internal factors. Solvents that form solvates include, but are not limited to, water, acetone, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, isopropanol, and the like. A "hydrate" refers to an association or crystal formed when the solvent molecule is water, i.e., a compound bound by non-covalent intermolecular forces to stoichiometric or non-stoichiometric amounts of water.

[0052] The precursor compounds of the hexahydro-β-acid component compounds provided herein can be prepared by salting out to obtain a solid material with higher chemical purity and lower impurity content. The salting out method utilizes the principles of acid-base neutralization, acid-base coordination, or acid-base chelation to precipitate a salt of an amino acid derivative with a corresponding organic base, inorganic base, organic acid, or inorganic acid to obtain a feed-acceptable salt. The inorganic acid includes, but is not limited to, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, nitric acid, or a combination 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, magnesium hydroxide, or calcium hydroxide.

[0053] Feed-acceptable salts are salts of precursor compounds of the hexahydro-β-acid component compounds of the present invention formed with organic bases, inorganic bases, organic acids, or inorganic acids that are non-toxic to animals. The term "feed-acceptable" means that the substance or composition must be chemically or toxicologically suitable for use in feed or for the intended animal.

[0054] In some embodiments, post-treatment of the precursor compound of the hexahydro-β-acid component compound of the present invention further involves forming an acid-base coordination salt and / or an acid-base chelate salt with an inorganic acid or an organic acid during salting-out precipitation, wherein the organic acid includes but is not limited to acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, malic acid, 2-hydroxypropionic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, glucuronic acid, galacturonic acid, citric acid, tartaric acid, aspartic acid, glutamic acid, benzoic acid, p-toluic acid, cinnamic acid, p-toluenesulfonic acid, benzenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, trifluoromethanesulfonic acid, or a combination thereof.

[0055] Application of compounds

[0056] The precursor compound of the hexahydro-β-acid component compound provided by the present invention and its solvate or feed-acceptable salt are used in the preparation of animal feed additives.

[0057] The "animal" referred to in the present invention refers to a human or farmed animal that cannot synthesize organic matter from inorganic matter and can only use organic matter as food to carry out life activities such as eating, digestion, absorption, breathing, circulation, excretion, sensation, movement and reproduction. "Farmed animals" include poultry, livestock, aquaculture animals and other animals that are legally captured and artificially raised, including pets, such as cats and dogs. The term "livestock" refers to any of pigs, cattle, horses, goats, sheep, deer and many useful rodents. The term "poultry" includes, for example, chickens, ducks, geese, quails, pigeons, etc. The term "aquaculture animals" includes, for example, fish, shrimp, turtles, tortoises, etc.

[0058] The precursor compound of the hexahydro-β-acid component compound provided by the present invention and its solvate or feed-acceptable salt are used to prepare a non-nutritional additive for improving the production performance of animals at various growth stages. The animals can be selected from livestock, poultry, aquaculture animals or pets at various growth stages.

[0059] Furthermore, the livestock include but are not limited to pigs, cattle, sheep, horses, rabbits, minks or donkeys, the poultry include but are not limited to chickens, turkeys, ducks, geese, quails or pigeons, the aquaculture animals include but are not limited to fish, shrimp, turtles, crabs, tortoises, bullfrogs, eels or loaches, and the pets include but are not limited to dogs or cats of various subspecies.

[0060] In one embodiment, the precursor compound of the hexahydro-β-acid component compound provided by the present invention and its solvate or feed-acceptable salt are used to prepare a feed additive for improving the production performance of pigs, which has an improving effect on the feed intake, average daily weight gain or feed conversion rate of pigs.

[0061] In another embodiment, the feed additive prepared using the precursor compound of the hexahydro-β-acid component compound provided by the present invention and its solvate or feed-acceptable salt can significantly improve the production performance of broilers or laying hens.

[0062] In another embodiment, the precursor compound of the hexahydro-β-acid component compound provided by the present invention and its solvate or feed-acceptable salt is used to prepare a feed additive for improving the production performance of fish.

[0063] Feed composition involved in this application

[0064] A stable feed composition comprising at least one of a precursor compound of a hexahydro-β-acid component compound represented by formula (I), or a solvate thereof and a feed-acceptable salt thereof, and an optional feed-acceptable adjuvant, wherein the feed-acceptable adjuvant is selected from one or a combination of feed-acceptable carriers, diluents, excipients, and solvents.

[0065] The "composition" referred to in this application refers to a compound group containing one or more compounds as active ingredients.

[0066] The term "comprising" as used herein is an open-ended expression, encompassing the contents explicitly stated herein, but not excluding other aspects. However, it should be noted that the feed composition provided herein does not contain hexahydrolupulone or its salts or esters. In one or more embodiments, in addition to the precursor compound of the hexahydro-β-acid component compound represented by formula (I), no other hexahydro-β-acid component compounds or their salts or esters are contained (except for a small amount of other hexahydro-β-acid component compounds that are unavoidable as impurities).

[0067] As used herein, a "stable feedable composition" is a composition that has sufficient stability to allow production and maintains the integrity of the compounds for a sufficient period of time to be consumed by an animal for the purposes detailed herein.

[0068] The "carrier" involved in this application refers to a feedable substance that can carry active ingredients, improve their dispersibility, and has good chemical stability and adsorption properties, and is an organic carrier and an inorganic carrier. The organic carrier is a material rich in crude fiber, including but not limited to corn flour, corn cob flour, wheat bran, rice husk powder, defatted rice bran, bran, corn stalk powder or peanut shell powder. The inorganic carrier is a mineral, mainly divided into calcium salts and silicon oxides, used for the preparation of trace element premixes, including but not limited to calcium carbonate, silicates, vermiculite, zeolite or sepiolite.

[0069] The "diluent" referred to in this application refers to a substance that evenly distributes the additive raw materials in the material, diluting high-concentration additive raw materials into a low-concentration premix or premix. It can separate trace ingredients from each other, reduce the interaction between active ingredients, and increase the stability of the active ingredients without affecting the physicochemical properties of the relevant substances. It includes organic diluents and inorganic diluents. Organic diluents include but are not limited to corn flour, degermed corn flour, dextrose (glucose), sucrose, coarse wheat flour with bran, roasted soybean flour, corn gluten meal, etc.; inorganic diluents include but are not limited to limestone, monocalcium phosphate, shell powder, kaolin (white clay), salt or sodium sulfate.

[0070] The excipients include one or more selected from the group consisting of a wetting agent that induces the inherent viscosity of the substance, a binder that binds the substance together, a disintegrant that breaks the entire sheet of the substance into many small particles, a retention agent that reduces the friction between particles, and an anti-sticking agent that prevents the material from sticking, including but not limited to magnesium stearate, talc, vegetable oil, magnesium lauryl sulfate, starch, starch slurry, water, inorganic salts, dextrin or powdered sugar.

[0071] The “solvent” referred to in this application refers to the solvent required to dissolve or disperse a solid, including but not limited to water, glycerol or ethanol.

[0072] In some embodiments, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is a precursor compound of hexahydrolupulone, or a solvate thereof, or a feed-acceptable salt thereof.

[0073] In one embodiment, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is hexahydrolupulone monopropyl ester as shown in formula (I2-1), or a solvate thereof, or a feed-acceptable salt thereof.

[0074] In one embodiment, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is hexahydrolupulone monobutyl ester as shown in formula (I2-2), or a solvate thereof, or a feed-acceptable salt thereof.

[0075] In one embodiment, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is hexahydrolupulone dipropyl ester as shown in formula (I2-5), or a solvate thereof, or a feed-acceptable salt thereof.

[0076] In one embodiment, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is hexahydrolupulone dibutyl ester having a structure as shown in formula (I2-6), or a solvate thereof, or a feed-acceptable salt thereof.

[0077] In some embodiments, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is a precursor compound of hexahydrolupulone, or a solvate thereof, or a feed-acceptable salt thereof.

[0078] In one embodiment, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is hexahydrolupulone monopropyl ester as shown in formula (I2-3), or a solvate thereof, or a feed-acceptable salt thereof.

[0079] In one embodiment, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is hexahydro-β-lupulone monobutyl ester as shown in formula (I2-4), or a solvate thereof, or a feed-acceptable salt thereof.

[0080] In one embodiment, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is hexahydro-β-lupulone dipropyl ester as shown in formula (I2-7), or a solvate thereof, or a feed-acceptable salt thereof.

[0081] In one embodiment, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is hexahydro-β-lupulone dibutyl ester as shown in formula (I2-8), or a solvate thereof, or a feed-acceptable salt thereof.

[0082] In some embodiments, the feed composition comprises a hexahydro-β-acid component compound whose precursor compound is a hexahydrolupulone precursor compound, or a solvate thereof, or a feed acceptable salt thereof, in combination with a hexahydrolupulone precursor compound, or a solvate thereof, or a feed acceptable salt thereof.

[0083] Specifically, the combination of the hexahydrolupulone precursor compound and the hexahydrolupulone precursor compound includes but is not limited to a combination of hexahydrolupulone monopropyl ester and hexahydrolupulone monopropyl ester, a combination of hexahydrolupulone monobutyl ester and hexahydrolupulone monobutyl ester, a combination of hexahydrolupulone dipropyl ester and hexahydrolupulone mono- and di-esters, and a combination of hexahydrolupulone dibutyl ester and hexahydrolupulone dibutyl ester.

[0084] In some embodiments, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is a hexahydro-lupulone precursor compound, or a solvate thereof, or a feed-acceptable salt thereof, and when combined with a hexahydro-lupulone precursor compound, or a solvate thereof, or a feed-acceptable salt thereof, the mass of the hexahydro-lupulone precursor compound in the combination is 1 part, and the mass of the hexahydro-lupulone precursor compound is no more than 0.5 part and not less than 0.01 part.

[0085] In some embodiments, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is a hexahydro-lupulone precursor compound, or a solvate thereof, or a feed-acceptable salt thereof, and when combined with a hexahydro-lupulone precursor compound, or a solvate thereof, or a feed-acceptable salt thereof, the mass of the hexahydro-lupulone precursor compound in the combination is 1 part, and the mass of the hexahydro-lupulone precursor compound is 0.5 parts.

[0086] In some embodiments, the precursor compound of the hexahydro-β-acid component compound contained in the feed composition is a hexahydro-lupulone precursor compound, or a solvate thereof, or a feed-acceptable salt thereof, and when combined with a hexahydro-lupulone precursor compound, or a solvate thereof, or a feed-acceptable salt thereof, the mass of the hexahydro-lupulone precursor compound in the combination is 1 part, and the mass of the hexahydro-lupulone precursor compound is 0.25 parts.

[0087] In some embodiments, the above-mentioned feeding composition further comprises additional animal feed additives and / or animal feed raw materials.

[0088] The animal feed additive is a nutritional feed additive, a general feed additive or a medicinal feed additive.

[0089] The nutritional feed additives mentioned above refer to small or trace substances added to compound feed to balance feed nutrients, improve feed utilization, and directly exert nutritional effects on animals, including but not limited to amino acids, amino acid salts and their analogs, vitamins and vitamin-like substances, mineral elements and their complexes (chelates), microbial enzyme preparations or non-protein nitrogen.

[0090] The general feed additives mentioned above are also called non-nutritional additives, which refer to some non-nutritional substances added to feed to improve feed utilization, ensure feed quality and quality, and benefit animal health or metabolism, including but not limited to growth promoters, anthelmintics and health care agents, flavorings and attractants, feed conditioners, feed modulators, feed storage agents or Chinese herbal additives.

[0091] In some embodiments, the additional animal feed additives contained in the feeding composition are one or more of nutritional feed additives, general feed additives and medicinal feed additives.

[0092] More specifically, the non-nutritional additive is a growth promoter, including butyric acid, calcium butyrate, sodium butyrate, tannic acid, p-thymol, p-thymol ester, p-thymol salt, 2-hydroxybenzoic acid, benzoic acid or calcium benzoate, zinc oxide, zinc sulfate or zinc chloride.

[0093] In one embodiment, the non-nutritive additive is calcium butyrate.

[0094] In another embodiment, the non-nutritive additive is tannic acid.

[0095] More specifically, the medicinal feed additives include but are not limited to veterinary drug premixes that have the effects of preventing animal diseases and promoting animal growth and can be added to feed for a long term and mixed with carriers or diluents.

[0096] More specifically, the medicinal feed additive is a feed antibiotic, and the feed antibiotic includes but is not limited to polymyxin, salinomycin, avilamycin, bacitracin, virginiamycin, nosiheptide, flavomycin, enramycin, bethromycin, olaquinoxaline, oxytetracycline or chlortetracycline.

[0097] In some embodiments, the animal feed raw materials are grains and their processed products, oilseeds and their processed products, leguminous crop seeds and their processed products, tubers, roots and their processed products, other seeds, fruit products and their processed products, forage, roughage and their processed products, other plants, algae and their processed products, dairy products and their by-products, terrestrial animal products and their by-products, fish, other aquatic organisms and their by-products, minerals, microbial fermentation products and by-products, other feed raw materials and other feeding substances.

[0098] Use of feeding composition

[0099] The present application relates to the use of a stable feed composition comprising a precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable adjuvants.

[0100] In some embodiments, the above-mentioned stable feed composition comprising a precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable excipients is used to prepare an animal feed additive.

[0101] Furthermore, the animal feed additive is a feed additive that improves animal production performance, including but not limited to livestock feed additives, poultry feed additives, aquaculture animal feed additives or pet feed additives.

[0102] Specifically, the stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable adjuvants is used to prepare a livestock feed additive, wherein the livestock include but are not limited to pigs, cattle, sheep, horses, rabbits, minks, etc. at various growth stages.

[0103] Specifically, a stable feed composition comprising a precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and feed-acceptable adjuvants is used to prepare a poultry feed additive, wherein the poultry includes but is not limited to chickens, ducks, geese, pigeons, etc. at various growth stages.

[0104] Specifically, the above-mentioned stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable adjuvants is used to prepare aquaculture animal feed additives, wherein the aquaculture animals include but are not limited to fish, shrimp, crab, turtle, eel, etc. at various growth stages.

[0105] Specifically, the stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable excipients is used to prepare a pet feed additive, wherein the pet includes but is not limited to artificially raised dogs or cats.

[0106] In some embodiments, the animal feed additive prepared from the stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable excipients is a premix, a composite premix, an aqueous solution, or a granule.

[0107] In some embodiments, the stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable adjuvants is used to prepare animal feed.

[0108] The feed involved in this application refers to products that are industrially processed and produced for animal consumption.

[0109] Furthermore, the animal feed prepared using the stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable adjuvants is livestock feed, poultry feed, aquaculture animal feed, or pet feed.

[0110] Specifically, the stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable adjuvants is used to prepare livestock feed, wherein the livestock include but are not limited to pigs, cattle, sheep, horses, rabbits, minks, etc. at various growth stages.

[0111] Specifically, the stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable adjuvants is used to prepare poultry feed, wherein the poultry includes but is not limited to chickens, ducks, geese, pigeons, etc. at various growth stages.

[0112] Specifically, the above-mentioned stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or a solvate of the precursor compound and at least one feed-acceptable salt and optional feed-acceptable adjuvants is used to prepare feed for aquaculture animals, wherein the aquaculture animals include but are not limited to fish, shrimp, crab, turtle, eel, etc. at various growth stages.

[0113] Specifically, a stable feed composition comprising a precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable excipients is used to prepare pet feed, wherein the pets include but are not limited to artificially raised dogs or cats.

[0114] In some embodiments, the feed prepared from the stable feed composition comprising at least one of the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or a solvate thereof and a feed-acceptable salt, and optional feed-acceptable adjuvants is a single feed, a concentrated feed, a compound feed, a compound premix, or a concentrate supplement.

[0115] Specifically, the compound feed is a complete compound feed.

[0116] Methods for improving production performance of farmed animals

[0117] In some feeding embodiments, farmers administer the stable feeding composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and optional feed-acceptable adjuvants, or an animal feed additive prepared from the feeding composition, to animals together with feed, which can significantly improve the production performance of the animals.

[0118] Optionally, the feed composition is a feed additive premix, a feed additive compound premix, a granule or an aqueous solution, and is administered to animals together with feed.

[0119] In one embodiment, the feeding composition is a feed additive premix.

[0120] In one embodiment, the feed composition is a feed additive compound premix.

[0121] In some embodiments, the feed additive is in the form of a premix, a compound premix, a granule or an aqueous solution, which is mixed with animal feed and then consumed by the animal.

[0122] The animals are livestock, poultry, aquaculture animals or pets.

[0123] Specifically, the livestock include but are not limited to pigs, cattle, sheep, horses, rabbits, minks, etc. at all growth stages; the poultry include but are not limited to chickens, ducks, geese, pigeons, etc. at all growth stages; the aquaculture animals include but are not limited to fish, shrimps, crabs, turtles, eels, etc. at all growth stages; the pets include but are not limited to artificially raised dogs or cats.

[0124] In one embodiment, farmers administer a feed additive comprising at least one of a precursor compound of the hexahydro-β-acid component compound represented by formula (I), or a solvate thereof and a feed-acceptable salt thereof, together with feed to weaned pigs, thereby significantly improving the average daily weight gain rate and feed conversion rate of the weaned pigs.

[0125] In one embodiment, farmers administer the stable feed composition comprising the precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and feed-acceptable adjuvants, or the animal feed additive prepared from the feed composition, to broilers together with feed, thereby significantly reducing the feed-to-meat ratio of the broilers and improving the feed conversion rate.

[0126] In one embodiment, a farmer administers a stable feed composition comprising a precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and feed-acceptable excipients, or an animal feed additive prepared from the feed composition, to fish along with feed.

[0127] In one embodiment, a farmer administers a stable feed composition comprising a precursor compound of the hexahydro-β-acid component compound represented by formula (I), or at least one of its solvates and feed-acceptable salts, and feed-acceptable excipients, or an animal feed additive prepared from the feed composition, to a puppy along with feed.

[0128] In other feeding embodiments, farmers feed animals an animal feed prepared from a stable feed composition comprising a precursor compound of the hexahydro-β-acid component compound represented by formula (I), or a solvate thereof and at least one feed-acceptable salt, and feed-acceptable adjuvants, which can significantly improve the production performance of the animals.

[0129] Optionally, the feed composition is concentrated feed, compound feed, compound premix or concentrate supplement, and is directly given to animals as animal feed.

[0130] The feed additive premix refers to a uniform mixture composed mainly of any two or more types of nutritious feed additives among mineral trace elements, vitamins, microorganisms, and amino acids, and the precursor compound of the hexahydro-β-acid component compound provided in this application or other feed additives, carriers and (or) diluents prepared in a certain proportion, wherein the content of the nutritious feed additive can meet the basic nutritional needs of the specific physiological stage of the animal to which it is applicable, and the addition amount in the compound feed, concentrate supplement or animal drinking water is not less than 0.1% and not more than 10%.

[0131] The concentrated feed refers to feed mainly containing protein, minerals and feed additives prepared in a certain proportion.

[0132] The compound feed is a feed prepared by combining a variety of feed raw materials and feed additives in a certain proportion according to the nutritional needs of the farmed animals.

[0133] The concentrate supplement refers to a feed prepared by combining a variety of feed raw materials and feed additives in a certain proportion in order to supplement the nutrition of herbivorous animals.

[0134] In one embodiment, the feeding composition is a complete feed.

[0135] The embodiments of the present application will be described in detail below with reference to the examples, but it will be understood by those skilled in the art that the following examples are merely illustrative of the present application and should not be considered as limiting the scope of the present application. In the examples, if specific conditions are not specified, the conditions are carried out according to conventional conditions or manufacturer recommendations. The reagents or instruments used are not specified by the manufacturer and are conventional products that can be purchased commercially.

[0136] Example A Semi-synthetic Preparation of Precursor Compounds of Hexahydro-β-Acid Component Compounds

[0137] Those skilled in the art will recognize that other methods for preparing the hexahydro-β-acid component compounds of the present application are considered to be within the scope of the present application. For example, the synthesis of the non-exemplified hexahydro-β-acid component compounds according to the present application can be successfully completed by those skilled in the art through modification methods, such as appropriate protection of interfering groups, by utilizing other reagents or making some conventional modifications to the reaction conditions.

[0138] Preparation of A1β-acid

[0139] In a 5L beaker, add 1kg of hop extract, 3L of pure water, and 1L of ethanol in sequence. Stir to dissolve the extract. Add 0.5L of KOH solution dropwise to adjust the solution pH to 13. Let stand and filter to remove insoluble matter. Pass CO2 gas through the filtrate to adjust the pH to 8.5. Cool and let stand for 2 hours, then filter to obtain the crude product.

[0140] The crude product was dissolved in 200 mL of n-hexane and washed with water (150 mL × 3). The organic phase was concentrated to obtain a paste, and then 300 mL of KOH lye was added to adjust the pH to 12.5 for dissolution. The product was washed with n-hexane (200 mL × 3). The organic phase was discarded, and the aqueous phase was acidified to adjust the pH to 8.5. The product was then extracted with n-hexane (200 mL × 3). The organic layers were combined and dried over sodium sulfate pentahydrate. The organic layers were removed by concentration under reduced pressure to obtain 400 g of crystalline β-acid. HPLC content analysis results showed that the main components of the β-acid crystals were synlupione, galupione, and lupulone.

[0141] Preparation of A2 hexahydro-β-acid

[0142] Dissolve 50 g of β-acid in 300 mL of 95% ethanol, add 1.7 g of 10% Pd / C, replace the atmosphere with hydrogen, and stir at room temperature overnight. Monitor by HPLC until the reaction is complete. Filter the reaction mixture, and concentrate the filtrate to yield 46 g of hexahydro-β-acid crystals.

[0143] Isolation and purification of the main components of A3 hexahydro-β-acid

[0144] A3.1 Preparation of Hexahydrolupulone

[0145]

[0146] Hexahydro-β-acid was recrystallized from n-hexane to obtain hexahydrolupulone crystals with a purity of 98.6%. 1 HNMR (500MHz, DMSO-d6): δ (ppm) 3.94-4.01 (m, 1H), 2.39 (t, 2H), 1.78-1.82 (m, 4H), 1.47-1.51 (m, 1H) ,1.28-1.35(m,4H),1.11(q,6H),0.95-1.01(m,4H),0.91(d,18H); LC-MS(ESI,pos.ion)m / z:407[M+H] + .

[0147] A3.2 Preparation of Hexahydrolupulone

[0148]

[0149] Hexahydro-β-acid was separated by preparative chromatography to obtain hexahydrolupulone with a purity of 99.1%. 1HNMR (500MHz, DMSO-d6): δ (ppm) 3.86-3.90 (m, 1H), 2.39 (t, 2H), 1.80-1.83 (m, 4H), 1.59-1.60 (m, 2H) ,1.47-1.52(m,5H),1.11(d,3H),0.90-0.96(m,7H),0.86(d,18H); LC-MS(ESI,pos.ion)m / z:421[M+H] + .

[0150] A3.3 Preparation of Hexahydrolupulone

[0151]

[0152] Hexahydro-β-acid was separated by preparative chromatography to obtain hexahydrolupulone with a purity of 98.9%. 1 HNMR (500MHz, DMSO-d6): δ (ppm) 2.82-2.84 (m, 2H), 2.37-2.40 (m, 2H), 2.00-2.05 (m, 1H), 1.80-1.88 (m, 4H), 1.77-1.80 ( m,1H),1.34-1.51(q,2H),1.29-1.33(m,2H),0.90-0.96(m,4H),0.86-0.90(m,24H); LC-MS(ESI,pos.ion)m / z:421[M+H] + .

[0153] Preparation of precursor compounds of A4 hexahydro-β-acid component compounds

[0154] A4.1 Preparation of Hexahydrolupulone Monoethyl Ester

[0155]

[0156] 2.0 g (4.92 mmol, 1.0 eq) of hexahydrolupulone was dissolved in 10 mL of dichloromethane, and 0.59 g (5.90 mmol, 1.2 eq) of triethylamine was added. The mixture was stirred and cooled to -5°C to 0°C. 0.47 g (5.90 mmol, 1.2 eq) of acetyl chloride was dissolved in 2 mL of dichloromethane and slowly added dropwise to the reaction system. After completion of the addition, the reaction system was warmed to room temperature and monitored by thin-layer chromatography until the reaction reached endpoint. 5 mL of pure water was added to the reaction system, stirred, and the organic phase was separated. The organic phase was washed with water (15 mL x 3), dried over sodium sulfate pentahydrate, and concentrated under reduced pressure to remove the organic solvent to obtain 1.35 g of the oily product in a yield of 61.3%.

[0157] A4.2 Preparation of Hexahydrolupulone Monopropyl Ester

[0158]

[0159] 2.0 g (4.92 mmol, 1.0 eq) of hexahydrolupulone was dissolved in 10 mL of dichloromethane, and 0.48 g (5.90 mmol, 1.2 eq) of triethylamine was added. The mixture was stirred and cooled to -5°C to 0°C. 0.55 g (5.90 mmol, 1.2 eq) of propionyl chloride was dissolved in 3 mL of dichloromethane and slowly added dropwise to the reaction system. After completion of the addition, the reaction system was warmed to room temperature and monitored by thin-layer chromatography until the reaction reached endpoint. 5 mL of pure water was added to the reaction system, stirred, and the organic phase was separated. The organic phase was washed with water (15 mL x 3), dried over sodium sulfate pentahydrate, and concentrated under reduced pressure to remove the organic solvent to obtain 1.52 g of the oily product in a yield of 66.8%.

[0160] A4.3 Preparation of Hexahydrolupulone Monobutyl Ester

[0161]

[0162] 2.0 g (4.92 mmol, 1.0 eq) of hexahydrolupulone was dissolved in 10 mL of dichloromethane, and 0.60 g (5.90 mmol, 1.2 eq) of triethylamine was added. The mixture was stirred and cooled to -5°C to 0°C. 0.62 g (5.90 mmol, 1.2 eq) of butyryl chloride was dissolved in 3 mL of dichloromethane and slowly added dropwise to the reaction system. After completion of the addition, the reaction system was warmed to room temperature and monitored by thin-layer chromatography until the reaction reached endpoint. 5 mL of pure water was added to the reaction system, stirred, and the organic phase was separated. The organic phase was washed with water (15 mL x 3), dried over sodium sulfate pentahydrate, and concentrated under reduced pressure to remove the organic solvent to obtain 1.29 g of the oily product in a yield of 55.1%.

[0163] A4.4 Preparation of Hexahydrolupulone Dipropyl Ester

[0164]

[0165] 2.5 g (6.15 mmol, 1.0 eq) of hexahydrolupulone was placed in a 50 mL dry reaction vessel, purged with nitrogen to remove oxygen and moisture, and then 20 mL of n-heptane and 0.1 g (0.82 mmol, 0.13 eq) of 4-lutidine were added sequentially. The reaction system was cooled to 0°C, and 1.37 g (14.76 mmol, 2.4 eq) of propionyl chloride and 1.49 g (14.76 mmol, 2.4 eq) of triethylamine were added dropwise. After the additions were completed, the reaction system was stirred and monitored by thin-layer chromatography until the reaction was complete. The reaction solution was filtered to remove triethylamine hydrochloride and then concentrated under reduced pressure to remove the solvent to obtain a crude yellow oil. The product was separated by silica gel column chromatography (dichloromethane / methanol (w / w) = 10 / 1) to obtain 1.39 g of hexahydrolupulone dipropyl ester in a yield of 43.7%.

[0166] A4.5 Preparation of Hexahydrolupulone Dibutyl Ester

[0167]

[0168] 2.5 g (6.15 mmol, 1.0 eq) of hexahydrolupulone was placed in a 50 mL dry reaction vessel, purged with nitrogen to remove oxygen and moisture, and then 20 mL of n-heptane and 0.1 g (0.82 mmol, 0.13 eq) of 4-lutidine were added sequentially. The reaction system was cooled to 0°C, and 1.57 g (14.76 mmol, 2.4 eq) of butyryl chloride and 1.49 g (14.76 mmol, 2.4 eq) of triethylamine were added dropwise. After the additions were completed, the reaction system was stirred continuously and monitored by thin-layer chromatography until the reaction was complete. The reaction solution was filtered to remove triethylamine hydrochloride and then concentrated under reduced pressure to remove the solvent to obtain a crude yellow oil. The product was separated by silica gel column chromatography (dichloromethane / methanol (w / w) = 10 / 1) to obtain 1.59 g of hexahydrolupulone dibutyl ester in a yield of 47.2%.

[0169] A4.6 Preparation of Hexahydrolupulone Monopropyl Ester

[0170]

[0171] Dissolve 2.5 g (5.94 mmol, 1.0 eq) of hexahydrolupulone in 15 mL of dichloromethane, add 0.72 g (7.13 mmol, 1.2 eq) of triethylamine, and stir. Cool to -5°C to 0°C. Dissolve 0.66 g (7.13 mmol, 1.2 eq) of propionyl chloride in 6 mL of dichloromethane and slowly add dropwise to the reaction system. After completion of the addition, warm the reaction system to room temperature and monitor the reaction by thin-layer chromatography until endpoint. Add 5 mL of pure water to the reaction system, stir, and separate the organic phase. The organic phase is washed with water (20 mL x 3), dried over sodium sulfate pentahydrate, and concentrated under reduced pressure to remove the organic solvent, yielding 1.66 g of the oily product in a yield of 58.5%.

[0172] A4.7 Preparation of Hexahydrolupulone Monobutyl Ester

[0173]

[0174] Dissolve 2.5g (5.94mmol, 1.0eq) of hexahydrolupulone in 15mL of dichloromethane, add 0.72g (7.13mmol, 1.2eq) of triethylamine, and stir. Cool to -5°C to 0°C. Dissolve 0.76g (7.13mmol, 1.2eq) of butyryl chloride in 8mL of dichloromethane and slowly add dropwise to the reaction system. After completion of the addition, warm the reaction system to room temperature and monitor the reaction by thin-layer chromatography until endpoint. Add 6mL of pure water to the reaction system, stir, and separate the organic phase. The organic phase is washed with water (30mL x 3), dried over sodium sulfate pentahydrate, and concentrated under reduced pressure to remove the organic solvent to yield 1.45g of the oily product in a yield of 49.6%.

[0175] A4.8 Preparation of Hexahydrolupulone Dipropyl Ester

[0176]

[0177] 2.5 g (5.94 mmol, 1.0 eq) of hexahydrolupulone was placed in a 50 mL dry reaction vessel, purged with nitrogen to remove oxygen and moisture, and then 25 mL of n-heptane and 0.15 g (0.82 mmol, 0.14 eq) of 4-lutidine were added sequentially. The reaction system was cooled to 0°C, and 1.32 g (14.26 mmol, 2.4 eq) of propionyl chloride and 1.44 g (14.26 mmol, 2.4 eq) of triethylamine were added dropwise. After the additions were complete, the reaction system was stirred continuously and monitored by thin-layer chromatography until the reaction was complete. The reaction solution was filtered to remove triethylamine hydrochloride and then concentrated under reduced pressure to remove the solvent to obtain a crude yellow oil. The product was separated by silica gel column chromatography (dichloromethane / methanol (w / w) = 10 / 1) to obtain 1.42 g of hexahydrolupulone dipropyl ester in a yield of 44.9%.

[0178] A4.9 Preparation of Hexahydrolupulone Dibutyl Ester

[0179]

[0180] 2.5 g (5.94 mmol, 1.0 eq) of hexahydrolupulone was placed in a 50 mL dry reaction vessel, purged with nitrogen to remove oxygen and moisture, and then 25 mL of n-heptane and 0.1 g (0.82 mmol, 0.14 eq) of 4-lutidine were added sequentially. The reaction system was cooled to 0°C, and 1.52 g (14.26 mmol, 2.4 eq) of butyryl chloride and 1.44 g (14.26 mmol, 2.4 eq) of triethylamine were added dropwise. After the additions were complete, the reaction system was stirred and monitored by thin-layer chromatography until the reaction was complete. The reaction solution was filtered to remove triethylamine hydrochloride and then concentrated under reduced pressure to remove the solvent to obtain a crude yellow oil. The product was separated by silica gel column chromatography (dichloromethane / methanol (w / w) = 10 / 1) to obtain 1.72 g of hexahydrolupulone dibutyl ester in a yield of 51.7%.

[0181] A4.10 Preparation of Hexahydrolupulone Monodecyl Ester

[0182]

[0183] 2.5 g (6.15 mmol, 1.0 eq) of hexahydrolupulone was dissolved in 10 mL of dichloromethane, and 0.75 g (7.38 mmol, 1.2 eq) of triethylamine was added with stirring. The mixture was cooled to -5°C to 0°C. 1.41 g (7.38 mmol, 1.2 eq) of decanoyl chloride was dissolved in 8 mL of dichloromethane and slowly added dropwise to the reaction system. After completion of the addition, the reaction system was warmed to room temperature and monitored by thin-layer chromatography until the reaction reached endpoint. 5 mL of pure water was added to the reaction system, stirred, and the organic phase was separated. The organic phase was washed with water (15 mL x 3), dried over sodium sulfate pentahydrate, decolorized over silica gel, and concentrated under reduced pressure to remove the organic solvent to obtain 1.87 g of the oily product in a yield of 54.2%.

[0184] A4.11 Preparation of Hexahydrolupulone Monolaurate

[0185]

[0186] 2.5 g (6.15 mmol, 1.0 eq) of hexahydrolupulone was dissolved in 10 mL of dichloromethane, and 0.75 g (7.38 mmol, 1.2 eq) of triethylamine was added with stirring. The mixture was cooled to -5°C to 0°C. 1.61 g (7.38 mmol, 1.2 eq) of lauroyl chloride was dissolved in 8 mL of dichloromethane and slowly added dropwise to the reaction system. After completion of the addition, the reaction system was warmed to room temperature and monitored by thin-layer chromatography until the reaction was complete. 5 mL of pure water was added to the reaction system, stirred, and the organic phase was separated. The organic phase was washed with water (15 mL x 3), dried over sodium sulfate pentahydrate, and concentrated under reduced pressure to remove the organic solvent. The resulting crude product was separated by silica gel column chromatography (dichloromethane / methanol (w / w) = 10 / 0.8) to obtain 2.00 g of the oily product in a yield of 55.3%.

[0187] Note: The preparation method of hexahydrolupulone monopentyl ester and hexahydrolupulone monohexyl ester is the same as that of hexahydrolupulone monobutyl ester.

[0188] Example B Preparation of a feed composition of a precursor compound of a hexahydro-β-acid component compound

[0189] The feed composition of the present invention contains a precursor compound of the hexahydro-β-acid component compound at a content of greater than or equal to 0.00001%. The content can be adjusted based on the growth stages of different animals or the usability of feed industry products (e.g., feed additives, feed additive raw materials, etc.). The content can also be adjusted based on the specific weights of other nutrients and non-nutrients in different feed formulations to meet the needs of the feed formulation. The feed composition of the present invention will be explained below using a basic granular premix as an example. Any substitution or addition of similar formulations or formulation components without synergistic effects is considered consistent with the present invention.

[0190] Preparation method of feed composition: put raw materials and auxiliary materials into a mixing unit and mix them evenly, and put them into a granulator with a mass fraction of 1.3% hydroxypropyl methylcellulose aqueous solution (binder) at a ratio of 100:35, start mixing and cutting for 3-5 minutes, and after granulation, enter the fluidized bed for drying and pass through a 16-mesh screen after 30 minutes.

[0191] Raw materials: Precursor compound of the hexahydro-β-acid component compound prepared in Example A.

[0192] Auxiliary material (carrier): corn starch.

[0193] Product formula: as described in Table 1.

[0194] Table 1 Formula of mixed granular feed additive

[0195]

[0196]

[0197] Example C Study on the Thermal and Light Stability of the Precursor Compounds of the Hexahydro-β-Acid Component Compound

[0198] 1. Test materials

[0199] Experimental instruments and reagents: drug stability test chamber, high performance liquid chromatography (HPLC); methanol (chromatographic grade), phosphoric acid (analytical grade).

[0200] Test samples: hexahydrolupulone raw material, hexahydrolupulone monoethyl ester raw material, hexahydrolupulone monopropyl ester raw material, hexahydrolupulone monobutyl ester raw material.

[0201] 2 Experimental steps

[0202] 2.1 Sample preparation and experimental methods

[0203] Preparation of 1% premix sample of the test sample: Weigh 1g of the test sample raw material, add 99g of corn cob powder, and mix in a blender to obtain a 1% premix sample of the test sample for testing.

[0204] Stability test at 60°C: Samples of the test product and its 1% premix were placed in a Petri dish, spread into a thin layer ≤5 mm, and incubated at 60°C. Samples were taken on the 5th and 10th days for HPLC analysis. Each sample was sampled in duplicate. The test results are shown in Table 2.

[0205] Stability test under light: The test sample and its 1% premix were placed in a Petri dish and spread into a thin layer ≤5 mm. The samples were then placed in a light environment at 4500 Lx. Samples were taken for HPLC analysis on the 5th and 10th day. Each sample was sampled in triplicate. The test results are shown in Table 2.

[0206] 2.2 Preparation of standard solution

[0207] Accurately weigh 25.0 mg of the test sample, add an appropriate amount of methanol, and ultrasonically dissolve. Dilute to 25 mL in a volumetric flask to prepare a working stock solution. Take an appropriate amount of the working stock solution and dilute it with methanol to concentrations of 100 ppm, 500 ppm, and 1000 ppm, respectively. Filter through a 0.22 μm organic filter membrane, and analyze by HPLC. Verify the linearity between the sample concentration and the HPLC peak area response value, and plot a standard curve.

[0208] 2.3 Preparation of test solution

[0209] Accurately weigh 25.0 mg of the test sample and 2.0000 g of 1% test sample premix placed under different environmental conditions, accurately add 25 mL of methanol and ultrasonically dissolve for 10 min. After filtering through a 0.22 μm organic filter membrane, the samples were detected and analyzed by HPLC.

[0210] 2.4 HPLC detection conditions

[0211] Chromatographic column: Waters Symmetry C18 column (250 mm*4.6 mm, 5 μm); mobile phase: 0.02% phosphoric acid: methanol = 5:95 (v:v); detection wavelength: 235 nm; column temperature: 25°C; injection volume: 10 μL; flow rate: 1 ml / min.

[0212] 3 Experimental results

[0213] The results in Table 2 show that the active ingredient content of a 1% premix of hexahydrolupulone varied by more than 5% during the 10-day test period of the 60°C high-temperature stability test. In the light stability test, the active ingredient content varied by more than 15% on day 10. Furthermore, the active ingredient content of the monoethyl, monopropyl, and monobutyl esters of hexahydrolupulone, whether in the raw material or in the premix, did not vary by more than 5% during both the high-temperature and light tests. This indicates that short-chain fatty acid ester derivatives of hexahydrolupulone possess both thermal and light stability characteristics that meet the requirements for feed additives.

[0214] Table 2 Results of research on factors affecting the stability of hexahydrolupulone ester derivatives and their 1% premix

[0215]

[0216]

[0217] Example D Study on the Long-term Stability of the Precursor Compound of the Hexahydro-β-Acid Component Compound in Feed

[0218] 1. Test materials

[0219] Experimental instruments and reagents: drug stability test chamber, high performance liquid chromatography (HPLC); methanol (chromatographic grade), phosphoric acid (analytical grade), n-hexane (chromatographic grade), isopropanol (chromatographic grade).

[0220] Test samples: Hexahydrolupulone raw material, hexahydrolupulone monoethyl ester raw material, hexahydrolupulone monopropyl ester raw material, hexahydrolupulone monobutyl ester raw material, hexahydrolupulone monopentyl ester raw material (provided by the Chemical Department of Guangzhou Yingsaite Co., Ltd.), hexahydrolupulone monohexyl ester raw material (provided by the Chemical Department of Guangzhou Yingsaite Co., Ltd.), hexahydrolupulone monolaurate raw material, hexahydrolupulone monophosphate disodium salt (provided by the Chemical Department of Guangzhou Yingsaite Co., Ltd.), hexahydrolupulone monophosphate diethyl ester (provided by the Chemical Department of Guangzhou Yingsaite Co., Ltd.).

[0221] Feed: Master Zhu's premixed feed (Master Zhu's Good Milk Sutra 5.33% piglet compound premixed feed A: Master Zhu's Good Milk Sutra 2.67% piglet compound premixed feed B = 2:1 crushed and mixed), Zhengda suckling piglet starter feed, Inset broiler feed, Guangdong Kebang 4% premixed feed.

[0222] 2 Experimental steps

[0223] 2.1 Sample preparation and experimental methods

[0224] Preparation of 1% premix sample of the test sample: weigh 1g of the test sample raw material, add 99g of corn cob powder, and mix in a blender to obtain a 1% premix sample of the test sample.

[0225] Preparation of 500ppm test sample feed: dilute the 1% premix sample of the test sample with Master Zhu's premix feed step by step to 5000ppm and 500ppm test sample feed sample for testing.

[0226] Preparation of 100ppm test sample feed: dilute 1% premix sample of the test sample with Zhengda suckling pig creep feed step by step to 1000ppm and 100ppm test sample feed sample for testing.

[0227] Preparation of 100ppm test sample feed: 1% premix sample of the test sample was diluted step by step with broiler feed provided by Nanxiong to 1000ppm and 100ppm test sample feed samples for testing.

[0228] Preparation of 2000ppm test sample feed: dilute the 1% premix sample of the test sample with Guangdong Kebang 4% premix feed in stages to 5000ppm and 2000ppm test sample feed samples for testing.

[0229] Experimental Method: Three parallel batches of the above-mentioned test feed samples were tested at 25°C ± 2°C and RH 60% ± 10%. Samples were collected for HPLC analysis on days 5, 10, 15, 30, 60, and 90. The test results are shown in Table 3.

[0230] 2.2 Preparation of standard solution

[0231] Accurately weigh 25.0 mg of the test sample, dissolve it in an appropriate amount of methanol or n-hexane by ultrasonication, and dilute to 50 mL in a volumetric flask to prepare a working stock solution with a mass concentration of 500 ppm. Take an appropriate amount of the working stock solution and dilute it with methanol or n-hexane to concentrations of 10 ppm, 25 ppm, 50 ppm, 100 ppm, and 250 ppm, respectively. Filter through a 0.22 μm organic filter membrane, and analyze by HPLC. Verify the linearity between the sample concentration and the HPLC peak area response value, and plot a standard curve.

[0232] 2.3 Preparation of test solution

[0233] Accurately weigh 5.0000 g of the test feed samples taken at different sampling times, accurately add 25 mL of methanol or n-hexane and ultrasonically dissolve for 10 min, filter through a 0.22 μm organic filter membrane, and then analyze by HPLC.

[0234] 2.4 HPLC detection conditions

[0235] Reversed-phase chromatography: instrument: Waters E2695 PDA detector; mobile phase: 0.02% phosphoric acid: methanol = 5:95 (v:v); chromatographic column: Waters Symmetry C18 (250 mm*4.6 mm, 5 μm); detection wavelength: 235 nm; column temperature: 25°C; injection volume: 10 μL; flow rate: 1 ml / min.

[0236] Normal phase chromatography: instrument: Shimadzu LC-14C / SPD-15C; chromatographic column: SuperSil NH2 column, 5 μm 250 mm × 4.6 mm; mobile phase: n-hexane:isopropanol = 60:40 (v:v); detection wavelength: 235 nm; column temperature: 25°C; sample volume: 20 μL; flow rate: 1 ml / min.

[0237] 3 Experimental results

[0238] As can be seen from the results in Table 3, during the test period, the content of hexahydrolupulone in the feed decreased significantly after 30 days, and the content of the active ingredient of the phosphate series compounds of hexahydrolupulone in the feed decreased rapidly and significantly in the early stage of the test. However, the fatty acid ester derivatives of hexahydrolupulone can maintain good stability in the feed under room temperature conditions. Among them, hexahydrolupulone monoethyl ester, hexahydrolupulone monopentyl ester and hexahydrolupulone monohexyl ester have poor stability in some feeds.

[0239] Table 3 Results of the study on the room temperature stability of hexahydrolupulone ester derivatives in feed

[0240]

[0241]

[0242] Example E Effect of Hexahydro-β-Acid Component Precursor Compounds on Pig Production Performance

[0243] A total of 480 "Du Changda" lean hybrid piglets of similar weight at 67 days of age were randomly divided into 16 treatment groups, with 3 replicates in each group and 10 pigs in each replicate, half male and half female. The pig pens and equipment were disinfected before the experiment. The pigs were housed in separate pens under the same feeding and management conditions during the experimental period. During the experiment, the experimental pigs had free access to food and water and were fed twice a day. The experimental groups were the control group (group 1) and experimental groups 2 to 16. Among them, the control group was given only the basal diet, and experimental groups 2 to 16 were given a diet supplemented with a hexahydro-β-acid component compound or a precursor compound of a hexahydro-β-acid component compound in the form of a mixture on the basis of the basal diet, and the concentration of the active ingredient of each additive in experimental groups 2 to 16 was 5 ppm or 25 ppm, as shown in Table 4.

[0244] No additional antioxidants or growth promoters were added to the experimental groups during the entire feeding process. The experimental period was 14 days. Taking each replicate as a unit, the animals were weighed on the 14th day without water or feed for 12 hours. The average daily feed intake (ADFI, g / d*head), average daily weight gain (ADG, g / d*head) and feed-to-meat ratio (FCR) of each experimental group were calculated. The calculation formula is as follows:

[0245] Average daily feed intake = (total amount of ingredients - amount of leftover feed) / (number of experimental days × number of pigs per replicate);

[0246] Average daily weight gain = (average weight at the end of the experiment - average weight at the beginning of the experiment) / number of experimental days;

[0247] Feed-to-meat ratio = average daily feed intake / average daily weight gain.

[0248] The test results are shown in Table 4.

[0249] Table 4 Effects of hexahydro-β-acid component compounds and their precursor compounds on the production performance of pigs

[0250]

[0251]

[0252] From the results in Table 4, it can be seen that this experiment compared and evaluated the effects of the test products on the production performance of the experimental pigs in terms of feed intake, weight gain and feed conversion rate. The ester derivatives of hexahydrolupulone or hexahydrolupulone increased the feed intake of the experimental pigs to varying degrees compared with the blank group or the hexahydrolupulone group or the hexahydrolupulone group. Moreover, at the same dosage, the effects of the monoethyl ester, monopropyl ester or monobutyl ester of hexahydrolupulone or hexahydrolupulone on the feed-to-meat ratio of the experimental pigs were equivalent to or better than those of hexahydrolupulone or hexahydrolupulone, respectively. However, the effects of the monohexyl ester, monodecyl ester and monolauric ester of hexahydrolupulone were not as significant as those of the monoethyl ester and monobutyl ester. The possible reason is that these ester compounds were not completely hydrolyzed or the hydrolysis rate was too slow after entering the animal intestine.

[0253] Example F Effect of Hexahydro-β-Acid Component Precursor Compounds on Broiler Performance

[0254] The experiment used a single-factor randomized design. A total of 540 one-day-old, similar-weight, three-yellow-feathered broilers with an average weight of 50g were randomly divided into six treatment groups, each with six replicates, half male and half female, and 15 three-yellow-feathered broilers per replicate. The chicken coop and equipment were disinfected before the experiment. During the experimental period, the chickens were caged in the same coop under the same feeding and management conditions. The basal diet consisted mainly of corn-soybean meal, and no other antioxidants or growth promoters were added during the entire feeding process. The experimental groups were the control group (Group 1) and experimental groups 2 to 6. Among them, the control group was given only the basal diet, while experimental groups 2 to 6 were given diets supplemented with hexahydrolupulone, hexahydrolupulone monoethyl ester, and hexahydrolupulone monobutyl ester, respectively, on top of the basal diet, as shown in Table 5.

[0255] The experimental period lasted 30 days. The experimental chickens had free access to water and food and were fed twice daily. Each replicate was weighed at 31 days of age (without feeding for 12 hours, but with water), and the feed consumption of the experimental chickens was recorded. The average daily feed intake (ADFI, g / day per bird), average daily weight gain (ADG, g / day per bird), and feed-to-cooking ratio (FCR) of the experimental chickens in each group were calculated using the following formula:

[0256] Feed to weight ratio (FCR) = average daily feed intake / average daily gain.

[0257] The test results are shown in Table 5.

[0258] Table 5 Study on the application effect of hexahydro-β-acid component compound precursor compounds in broiler feed

[0259] Test group test sample ADFI (g / d*) ADG (g / d*) FCR1 group -49.55 23.14 2.142 group Hexahydrolupulone 20ppm 49.68 25.19 1.97 group Hexahydrolupulone monoethyl ester 2ppm 49.13 25.67 2.024 group Hexahydrolupulone monoethyl ester 20ppm 50.06 27.36 1.95 group Hexahydrolupulone monobutyl ester 2ppm 48.32 24.65 1.96 group Hexahydrolupulone monobutyl ester 20ppm 49.95 25.88 1.93

[0260] As shown in Table 5, this study compared and evaluated the effects of the test products on the production performance of experimental chickens in terms of feed intake, weight gain, and feed conversion rate. The monoethyl and monobutyl esters of hexahydrolupulone showed equivalent effects on improving the production performance of broiler chickens as hexahydrolupulone. Industrial Applicability

[0261] In the present application, the precursor compound of the hexahydro-β-acid component compound hexahydrolupulone and / or hexahydrolupulone has an effect of improving animal production performance that is basically equivalent to hexahydrolupulone and / or hexahydrolupulone. When used as a functional ingredient of the feed composition, it can not only avoid the problems of thermal stability and long-term storage stability of the hexahydro-β-acid component compound during the feed pelleting process, but also have the effect of improving the production performance of the fed animal that is equivalent to that of the hexahydro-β-acid component compound.

Claims

1. A precursor compound of a hexahydro-β-acid component compound having a structure as shown in formula (I), or a solvate of the precursor compound and a feed acceptable salt thereof: in, R 1 is selected from substituted or unsubstituted C 1 -C 2 Alkyl; R 2 or R 3 independently selected from H, substituted or unsubstituted straight or branched chain C 2 -C 4 Carbonyl.

2. The precursor compound of the hexahydro-β-acid component compound according to claim 1, It is characterized in that The R 2 or R 3 independently selected from H or C(=O)CH 2 CH 3 , and they are not H at the same time.

3. The precursor compound of the hexahydro-β-acid component compound according to claim 1, It is characterized in that The R 2 or R 3 independently selected from H or C(═O)(CH 2 ) 2 CH 3 , and they are not H at the same time.

4. The precursor compound of the hexahydro-β-acid component compound according to claim 1, It is characterized in that The precursor compound has one of the following structures:

5. A feeding composition, It is characterized in that The feed composition comprises a precursor compound of the hexahydro-β-acid component compound according to any one of claims 1 to 4, or at least one of a solvate and a feed-acceptable salt of the precursor compound, and optionally a feed-acceptable auxiliary material.

6. The feed composition according to claim 5, It is characterized in that The precursor compound of the hexahydro-β-acid component compound is any compound represented by formula (I2-1) or formula (I2-2):

7. The feeding composition according to claim 5, It is characterized in that The precursor compound of the hexahydro-β-acid component compound is any compound represented by formula (I2-3) or formula (I2-4):

8. The feed composition according to claim 5, It is characterized in that The precursor compound of the hexahydro-β-acid component compound is any compound represented by formula (I2-1) or formula (I2-3):

9. The feed composition according to claim 5, It is characterized in that The precursor compound of the hexahydro-β-acid component compound is any compound represented by formula (I2-2) or formula (I2-4):

10. The feed composition according to claim 5, It is characterized in that The feeding composition may further comprise additional animal feed additives and / or animal feed raw materials.

11. The feed composition according to claim 10, It is characterized in that The additional animal feed additives include one or more selected from the group consisting of nutritional feed additives, non-nutritional additives and medicinal feed additives.

12. Use of the precursor compound of the hexahydro-β-acid component compound according to any one of claims 1 to 4 or the feeding composition according to any one of claims 5 to 11 in the preparation of an animal feed additive.

13. Use of the precursor compound of the hexahydro-β-acid component compound according to any one of claims 1 to 4 or the feeding composition according to any one of claims 5 to 11 in the preparation of animal feed.

14. A method for improving animal production performance, It is characterized in that The method comprises: feeding an animal with the feeding composition according to any one of claims 5 to 11; or feeding an animal with a feed comprising the feeding composition according to any one of claims 5 to 11.

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