KR101484611B1 - Composition for improving milk protein synthesis containing at least vitamin A, vitamin C and their analogs, feed additive comprising the same and method for improving milk protein synthesis using the same - Google Patents

Abstract

 The present invention relates to a pharmaceutical composition comprising at least one of vitamin A, vitamin C and analogs thereof, preferably at least one of vitamin A, vitamin C and their analogs, A composition for promoting synthesis. Accordingly, the present invention can be applied as a feed additive to increase the milk protein of the milk obtained from the livestock fed the milk, thereby improving the milk quality. In addition, the content of vitamin A and vitamin C in the meat and milk of livestock to which the present invention is applied can be increased.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for enhancing milk protein synthesis comprising at least one of vitamin A, vitamin C and their analogues, a feed composition containing the same, and a method for enhancing milk protein synthesis using the same. C and their analogs, feed additive comprising the same and method for improving milk protein synthesis using the same}

The present invention relates to a composition for promoting synthesis of a milk protein, a method for producing the same, and a method for enhancing the synthesis of milk protein using the same.

Vitamins are not a component of the tissue or an energy source, but they are essential factors that regulate tissue synthesis and energy production and act as coenzyme. Because these vitamins are consumed through livestock products, they are often used as feed additives to increase the amount of vitamins contained in livestock products. In addition to the purpose of increasing the amount of vitamins contained in livestock products, the vitamins also contribute to the health of the cattle and improve the quality of the final product.

Vitamin A is known to have the effect of decreasing mastitis generation, decreasing somatic cell number, and improving fertility in cattle. In addition, vitamin C promotes the decomposition of stress hormones (cortisol), and it has effects such as high temperature stress in summer, overcoming of labor and environmental change stress, improvement of disease resistance, activation of lipid synthetic hormone (GPDH) have.

On the other hand, milk protein is a protein derived from milk. Milk protein has many amino acids which are insufficient in vegetable protein, such as lysine and methionine, which are essential amino acids. Although many attempts have been made to increase the content of such a milk protein in the past, there has been no effective method for increasing the content of milk protein yet.

The present invention provides a composition for increasing milk protein. In addition, a problem to be solved by the present invention is to provide a method for raising milk protein in an animal producing milk.

In order to solve the above-mentioned problems, the present invention relates to a nanosomes containing at least one of vitamin A, vitamin C and their analogs, preferably at least one of vitamin A, vitamin C and their analogues, As an active ingredient, a composition for enhancing milk protein synthesis.

The nanosome may be prepared by a mixture comprising a phospholipid and water, and the phospholipid may be any one selected from the group consisting of lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and dipalmitoylphosphaticholine.

The present invention also provides a feed additive for enhancing milk protein synthesis comprising the composition according to the present invention.

The present invention also encompasses nanosomes containing at least one of vitamin A, vitamin C and their analogs, preferably at least one of vitamin A, vitamin C and their analogs as an active ingredient A feed composition for enhancing milk protein synthesis.

The nanosome may be prepared by a mixture comprising a phospholipid and water, and the phospholipid may be any one selected from the group consisting of lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and dipalmitoylphosphaticholine.

The present invention relates to a method of enhancing the synthesis of milk proteins in mammary cells through nanosomes containing at least one of the active ingredients of at least one of vitamin A, vitamin C and their analogues, preferably vitamin A, vitamin C and their analogs . As a result, the milk protein content of milk obtained from livestock is increased, and milk quality can be improved. In addition, the nano-encapsulation of the active ingredient increases the absorption rate of the active ingredient, thereby promoting the physiological activity of the livestock and increasing the content of vitamin A and vitamin C in the meat or milk using livestock, thereby producing high quality meat and milk.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments of the invention and, together with the description of the invention, It should not be construed as limited.
FIG. 1 is a graph of the cell viability test result of the nanosome according to the present invention.
FIG. 2 is a graph showing the results of experiments on the expression level of bCSNB in the nanosome according to the present invention.
FIG. 3 is a graph showing the results of experiments on the expression level of bCSNK in the nanosome according to the present invention.

Hereinafter, the present invention will be described in detail. The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms and the inventor may appropriately define the concept of the term in order to best describe its invention It should be construed as meaning and concept consistent with the technical idea of the present invention. Therefore, the constitution described in the embodiments described herein is merely the most preferred embodiment, and does not represent all of the technical ideas of the present invention, so that various equivalents and variations And the like.

The present invention relates to a pharmaceutical composition comprising at least one of vitamin A, vitamin C and analogs thereof, preferably at least one of vitamin A, vitamin C and their analogs, A composition for promoting synthesis is provided.

In the present invention, milk protein is exemplified by milk protein as milk protein, and the composition according to the present invention can enhance the synthesis of milk protein, and such composition can be applied to feed additives to be fed to livestock. The feed additive according to the present invention is a feed additive to be applied to livestock producing dairy products such as dairy cattle, goat, goat and goat. The feed additive according to the present invention enhances the milk protein content of milk obtained from livestock .

The present inventors have found that the viability of mammary gland cells is improved by nano-somatization of at least one or more active ingredients of vitamin A, vitamin C and their analogues, preferably vitamin A, vitamin C and their analogs, And the amount of expression of the gene was found to increase. Based on this, it is possible to enhance the synthesis of milk protein by using nanosomes containing at least one of vitamin A, vitamin C and their analogues, preferably at least one of vitamin A, vitamin C and their analogues, And the present invention has been completed based on the fact that it can be used for a purpose. That is, the present invention relates to a composition using nanosomes containing at least one or more of vitamin A, vitamin C and their analogues, preferably at least one of the above-mentioned vitamin A, vitamin C and their analogs, and a feed additive comprising the same And the feed additive was applied to the livestock producing milk to further enhance the milk protein in the milk obtained from the livestock.

In addition, vitamin A or vitamin C is also increased in livestock meat and milk to which the feed additive comprising the composition according to the present invention is applied.

The composition according to the present invention comprises at least one of vitamin A, vitamin C and analogs thereof, at least one of vitamin A, vitamin C and their analogs. That is, vitamin A and vitamin C may be included or may be included in some modified forms which are derivatives of these vitamins. For example, it can be included as a vitamin ester, a vitamin ester salt and the like, more specifically, retinyl palmitate, retinyl acetate, retinyl propionate, vitamin C magnesium phosphate (ascorbyl 2-phosphate magnesium salt) Corn-2-phosphate 6-palmitate, and the like. The most suitable form of vitamin A in the present invention is vitamin A acetate, and the most suitable form of vitamin C is ascorbic acid.

The active ingredient is preferably used in an amount of 5 to 20% by weight, more preferably 5 to 15% by weight, and more preferably 10% by weight, based on the total weight of the mixture for preparing nanosome But not limited to,

The nanosomes are prepared by a mixture comprising a phospholipid, and water. Or a solvent other than the water for dissolving the phospholipid may be further added according to the properties of the phospholipid. For example, all kinds of solvents capable of dissolving phospholipids such as ethanol may be added, One, ethanol is preferred.

Nanosomes containing the active ingredient may be prepared by a mixture comprising phospholipids, active ingredients, water, and the like. Or a solvent other than the water for dissolving the phospholipid may be further added according to the properties of the phospholipid. For example, any kind of solvent capable of dissolving phospholipids such as ethanol may be added. Preferably, ethanol May be added and are not limited to these kinds.

Nanosome is a substance expressed in various ways by nanosome, micelle, liposome and the like, and is not limited to this name. Is a thermodynamically stable and homogeneous structure made up of amphiphilic, for example, low molecular weight materials having both a hydrophilic group and a hydrophobic group at the same time. The amphipathic substance in the aqueous solution is composed of a hydrophilic group of the amphipathic substance externally and a hydrophobic group of the amphiphilic substance to form a layer and form a myoshell as a spherical association. These my shells can be represented as capsules and their names can vary. In addition, the layer by the phospholipid may be a bilayer or a multilayer as well as a single layer, and the number of such phospholipid layers is not limited. Mycels or capsules made with such amphiphilic phospholipids have an average particle size of nanoparticle size, for example from 1 to 1000 nm.

The active ingredient may be contained in the center of the my shell or between the phospholipid layer and the layer. In addition to increasing the uptake rate of the active ingredient through the nano-somatization of these active ingredients, the present invention further enhances milk protein synthesis through nano-somatization of at least one of the active ingredients of vitamin A, vitamin C and their analogs .

In the present invention, the microshell having an average particle diameter of 1 to 500 nm is preferable, and the microshell is more preferably an average particle diameter of 10 to 400 nm. It is further preferable that the average shell diameter of the shell is 100 to 300 nm. This is because, in the case of the average particle size of the my shell, not only the stability of the my shell particle but also the ability of the active substance to penetrate and the bio-absorption can be increased.

As the phospholipid contained in the nanosome, various phospholipids known in the art can be used. Examples of the phospholipid include phospholipids such as lanolin such as egg yolk lecithin, soybean lecithin and hydrogenated lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, Choline, etc. may be used. The phospholipid is preferably 2 to 10% by weight based on the total weight of the mixture for preparing the nanosome.

The present invention may further comprise a polyol for stabilizing the nanosome with an emulsion stabilizing adjuvant in a mixture for making the nanosome. As the polyol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, methylpropanediol, isoprene glycol, pentylene glycol, erythritol, xylitol, sorbitol and the like can be used. Propylene glycol, and the like are more preferable. Such a polyol is preferably used in an amount of 2 to 15% by weight, and more preferably 5 to 10% by weight, based on the total weight of the mixture for preparing the nanosome containing the active ingredient.

The present invention may further comprise a surfactant for stabilizing the nanosome in the mixture for making the nanosome. As such a surfactant, any of those known in the art may be used, and for example, an anionic surfactant, a cationic surfactant, a positive surfactant or a nonionic surfactant may be used, and preferably an anionic surfactant Or a nonionic surfactant is used. Specific examples of anionic surfactants include alkyl acyl glutamates, alkyl phosphates, alkyl lactylates, dialkyl phosphates, alkyl lactates and trialkyl phosphates. Specific examples of nonionic surfactants include alkoxylated alkyl ethers, alkoxylated alkyl esters, alkyl polyglycosides, polyglyceryl esters and sugar esters. More preferably, the surfactant is diacetyl phosphate. The content of the surfactant is preferably 0.5 to 1.5% by weight, more preferably 0.8 to 1.0% by weight, based on the total weight of the water-containing mixture for preparing the nanosome containing the active ingredient.

The nanosome of the present invention can be extruded through a filter having a straight passage or a curved passage or homogenized through a homogenizer to have a reduced uniform size distribution and the prepared solution can be dewatered or lyophilized have.

In addition, the present invention relates to a method for producing a nano-scale skin by passing an active ingredient according to the present invention through a high pressure homogenizer or the like by passing a mixture containing at least one active ingredient of vitamin A, vitamin C and analogues thereof, To provide a method of making cotton.

The present invention also encompasses nanosomes containing at least one of vitamin A, vitamin C and their analogs, preferably at least one of vitamin A, vitamin C and their analogs as an active ingredient A method of enhancing milk protein using a composition comprising: That is, the present invention provides a method for increasing the content of milk protein in milk obtained by administering an effective amount of at least one of vitamin A, vitamin C, and their analogs to an animal (i.e., a livestock producing milk such as cow, goat and chlorine) .

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

≪ Preparation of nanosome containing active ingredient >

Example 1

Lecithin (Lipoid S from Pharmachem) and ethanol are mixed and dissolved, and vitamin A, the active ingredient, is mixed. After that, water was added and stirred until completely dissolved to prepare a mixture. The respective contents in the mixture were 5% by weight of lecithin, 10% by weight of ethanol, 10% by weight of active ingredient and 75% by weight of water. After that, the nanotubes were processed by using the MN400 of MICRONOX Co., Ltd., and the pressure was cycled three times at 1000 bar.

Example 2

The nanosomes were prepared in the same manner as in Example 1, except that vitamin C was used instead of vitamin A as an active ingredient.

<Bovine Derived Mammary Viability Test>

The cell line was a bovine-derived mammary gland cell line, MAC-T cell line.

The medium composition of MAC-T cells was DMEM medium containing 100 units / ml of penicillin, 100 ug / ml of streptomycin, 5 ug / ml of insulin, 50 ug / ml of gentamicin, 1 ug / ml of hydrocortisone and 10% of FBS.

This experiment was conducted to measure cell viability. Yellow tetrazolium salt MTT (MTT) was reduced from living cells to water-insoluble formazan crystals. The crystals were dissolved in DMSO And the amount of the generated crystal was measured by a spectroscopic method. As a result, as the number of living cells increases, the number of these crystals increases. Therefore, the viability of cells can be easily measured by this method.

First, MAC-T cells were plated on a 12-well plate at 2 x 10 &lt; ^{4 &} gt ;. Experimental Example 1-1) Vitamin A 100 mg / ml, Experimental Example 1-2) Vitamin C 100 mg / ml, Experimental Example 1-3) Example 1 Nanosome containing 100 mg / ml of vitamin A and Experimental Example 1- 4) Nanosomes containing 100 mg / ml of vitamin C of Example 2 were treated in each cell. MTT solution was then added to each well at a volume of 10% and placed in a 37 ° C CO ₂ incubator for 2 to 3 hours. Then, the medium was removed, and 500 μl of DMSO was added thereto. After thoroughly mixing with pipetting, absorbance was measured at 570 nm.

The results on cell viability are shown in Fig. 1, a MAC-T cell treated with Vitamin A nanosome or a Vitamin C nanosome containing a vitamin C contains a nanosome containing vitamin A or a nanosome containing vitamin C (MAC-T) cells (control), which are not treated with the control cells, that is, the viability of the mammary gland cells.

&Lt; Experimental Expression of mRNA Expression Related to Milk Protein Synthesis in MAC-T Cells &

Experimental Example  2-1

Example 1, which is a nanosome containing vitamin A in MAC-T cells, is reacted for 24 hours. Then, RNA is extracted by chloroform extraction and cDNA is synthesized using reverse transcriptase. The reaction was carried out at 25 ° C for 10 minutes, then at 48 ° C for 120 minutes, then at 95 ° C for 5 minutes.

Then, real-time PCR is performed using Primer and SYBR corresponding to the indicator. The reaction conditions are annealing at 95 ° C for 40 cycles and at 60 ° C for 1 minute.

The primers used were as follows.

Experimental Example  2-2

In Experimental Example 2-2, Example 2, which is a nanosome containing vitamin A, was reacted, instead of reacting Example 1, which is a nanosome containing vitamin A.

Experimental Example  2-3

The same was done except that the nanosome containing the vitamin A or vitamin C was reacted.

 The expression level of bCSNB, which is one of the synthetic genes of β-casein in milk protein, is shown in FIG. 2, and the expression level of bCSNK, which is one of synthetic genes of κ-Caxein, is shown in FIG. 2 and 3, nanosome containing vitamin A and nanosome containing vitamin C are caused to express more genes for synthesizing milk protein.

Claims (9)

Vitamin A, vitamin C or a mixture thereof as an active ingredient,
Wherein the active ingredient is present in an amount of 5 to 20 wt% based on the total weight of the mixture for nanosome preparation. delete The method according to claim 1,
Wherein the nanosome is produced by a mixture comprising a phospholipid and water. The method of claim 3,
Wherein the phospholipid is at least one selected from the group consisting of lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and dipalmitoylphosphaticholine. A feed additive for enhancing milk protein containing nanosomes according to any one of claims 1 and 3 to 4. Vitamin A, vitamin C or a mixture thereof as an active ingredient,
Wherein the active ingredient is administered to the animal in an effective amount of nanosome comprising from 5 to 20% by weight based on the total weight of the mixture for nanosome preparation. delete The method according to claim 6,
Wherein said nanosome is produced by a mixture comprising a phospholipid and water. 9. The method of claim 8,
Wherein the phospholipid is at least one selected from the group consisting of lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and dipalmitoylphosphaticholine.

Images