KR20210044382A - Composition for preventing or treating osteoporosis comprising a milk derived exosome - Google Patents

Abstract

The present invention relates to a composition for preventing, treating or alleviating osteoporosis comprising exosomes derived from bovine colostrum as an active component. The exosome can inhibit the differentiation of osteoclasts, and thus can increase the bone density and bone mass percentage of the femur. In addition, since it is possible to change the intestinal microflora increased or decreased by the osteoporosis, the composition can be usefully used as a pharmaceutical or health functional food for the prevention, treatment or alleviation of osteoporosis.

Description

Composition for preventing or treating osteoporosis containing milk-derived exosomes {COMPOSITION FOR PREVENTING OR TREATING OSTEOPOROSIS COMPRISING A MILK DERIVED EXOSOME}

The present invention relates to a composition for preventing or treating osteoporosis comprising milk-derived exosomes, in particular, exosomes isolated from cow colostrum as an active ingredient.

Osteoporosis is the most common degenerative disease in developed countries as the number of older adults increases, and it may occur in young people depending on nutritional status and medications. Since degenerative diseases lower the quality of life due to depression, etc., prevention through proper exercise and nutrition is important.

Osteoporosis is known to be a skeletal disorder with reduced bone strength, and the prevalence of osteoporosis is high in women aged 50 to 69 years. In particular, osteoporotic fractures are most common among women and the elderly after menopause, and are a major cause of global health consumption. Although many biological, nutritional and behavioral factors contribute to the osteoporotic process, bone mass loss, structural deterioration and fall frequency eventually increase. Dairy supplements are widely recommended for the prevention of osteoporosis and subsequent fractures, because of the various functional gradients, including milk-based protein, casein phosphopeptide, and lactoferrin, which are beneficial for bone health. In particular, colostrum has been extensively studied as milk that is rich in nutrients produced in the mammary gland during the end of pregnancy and contains immune, developmental and tissue repair factors. Several studies have shown that colostrum consumption is beneficial for bone health and is being studied as an important functional substance for bone health.

Exosomes are membrane vesicles (30-150 nm) of intracellular origin. These extracellular vesicles (EV) mediate cell-to-cell interactions by the transfer of various proteins, lipids and RNAs between different cell types. Exosomes have been detected in all biological fluids (blood, milk, saliva, urine, etc.) and are considered a new factor in cell-cell interaction. They are emerging as important biomarkers of human disease. According to some studies, proteins present in milk-derived exosomes have biological roles including immune regulation, cell growth and repair. Interestingly, exosomal mRNA can be transferred from mother to infant via breastfeeding, providing immune-related functions. Orally administered bovine milk-derived exosomes improved experimental arthritis symptoms in mice. Therefore, milk-derived exosomes have a beneficial effect and a wide range of functions not only for the elderly but also for infants. Fortunately, cow milk contains large amounts of exosomes in both mature milk and colostrum, suggesting that exosomes such as mRNA and protein and their contents can be a good source of nutrients for research to develop functional supplements.

Although the relationship between milk and osteoporosis is still controversial, developing preventive substances from milk to prevent osteoporosis can contribute to social and medical cost savings, so milk-derived extracellular vesicles are advantageous in inhibiting osteoclast differentiation. There is a need for research on whether it can play a role and whether it can improve bone health in osteoporotic mice.

The present invention is to provide a pharmaceutical composition for preventing or treating osteoporosis, including exosomes isolated from bovine colostrum as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

The present invention provides a pharmaceutical composition for preventing or treating osteoporosis comprising exosomes isolated from bovine colostrum as an active ingredient.

The exosomes can inhibit the differentiation of osteoclasts.

The exosomes may increase the bone density and percentage of bone mass of the femur.

The exosome may again reduce the intestinal microbes increased by the osteoporosis, and increase the intestinal microbes reduced by the osteoporosis again.

The increased gut microbes may belong to a permeation's cuticle (firmicutes) or di-less Perry bacterium belongs to (deferribacteres), wherein the reduce intestinal microorganisms are Lactobacillus bacteria (Lactobacillus) or the thin steroid (Bacteroides).

The daily dosage of the composition may be 0.1 mg/kg to 10 mg/kg.

In one embodiment of the present invention, it provides a health functional food for preventing or improving osteoporosis comprising exosomes isolated from bovine colostrum as an active ingredient.

The present invention relates to a composition for preventing, treating or improving osteoporosis comprising exosomes isolated from bovine colostrum as an active ingredient, wherein the exosomes can inhibit the differentiation of osteoclasts, and increase the bone density and bone mass percentage of the femur I can make it. In addition, since it can change the intestinal microflora increased or decreased by the osteoporosis, it can be usefully used as a medicine or health functional food for preventing, treating or improving osteoporosis.

1 shows the effect of BCE treatment in vitro and in vivo: (A) Representative images of TRAP-positive cells in culture. As indicated by TRAP staining (multinucleated and expanded cells = TRAP-positive cells), differentiation stimulators (50 ng/ml of RANKL and 15 ng/ml of M- CSF) was added. (B) Schematic diagram of the experimental design using mice. Using the method described in Thiele et al., 2014 with slight modifications, the sustained-release pellet (7.5 mg prednisolone) was placed subcutaneously in the shoulder area of the rat to induce osteoporosis through glucocorticoid treatment. (C), (D) In Shame group mice, in H-BCE group mice supplied with high concentration of BCE (1.5 mg/Kg), in PDS (osteoporosis-induced) group mice, low concentration of BCE (0.15 mg/Kg) In PDS+L-BCE group mice fed with BCE (1.5mg/Kg) and in PDS+H-BCE group mice fed with high concentration of BCE (1.5mg/Kg), bone density and bone mass percentage of the left femur by X-ray micro-CT (D) Analysis. Results presented are the mean ± standard deviation (SD) of 4 mice/group. (E) Representative micro-CT images of the trabeculae of the femur for each group at a pixel size of 17.57 μm. Asterisks indicate significant differences between groups (*p <0.05, ** p <0.001).
Figure 2 shows the changes in the intestinal microflora induced by BCE uptake in mice with osteoporosis: (A) Two-dimensional main coordinate analysis based on unweighted Unifrac with clusters for each location subjected to different treatments (representative three groups) (PCoA). (B) Comparison of alpha diversity indices between the three groups. The Shannon index and Chao-1 were used to measure the diversity of the intestinal microbial community. (C) Cluster bar graph analysis showed cluster composition and species abundance in the three groups. (D) The composition of microorganisms in the genus by heat map analysis in three groups at the genus level. (E), (F) The abundance of Lactobacillus and Bacteriodes was divided into three representative groups. Asterisks indicate significant differences between groups (*p <0.05, ** p <0.001).

Osteoporosis is a systemic skeletal disease characterized by low bone mass and microstructural deterioration of bone tissue, resulting in increased bone fragility and susceptibility to fracture. In an aging society where life expectancy is increasing, the incidence of osteoporosis also increases rapidly. It has been suggested that inadequate nutritional intake can negatively affect bone metabolism. Recently, many studies have been made on the functionality of milk-derived exosomes, which play an important role in cell-cell interactions. However, there are only a few reports on the effect on the proliferation and differentiation of osteoblasts in vitro. Accordingly, the present inventors determined whether bovine colostrum-derived exosomes promote anti-osteoporosis in vitro and in vivo. In Raw264.7 cells treated with exosomes undergoing differentiation into osteoclasts, TRAP (tatarate resistant acidic phosphatase) stained cells were significantly inhibited. After oral administration of exosomes for 2 months, the present inventors induced osteoporosis in mice using glucocorticoid pellets. Interestingly, the bone mineral density (BMD) of the exosome-fed mouse group was significantly improved over that of the glucocorticoid-induced osteoporosis group without exosomes. In addition, Lactobacillus decreased in the intestinal microbial community of osteoporosis-induced mice, but the intestinal microbial community was effectively reversed by ingestion of exosomes.

Hereinafter, the present invention will be described in detail.

Pharmaceutical composition for preventing or treating osteoporosis

The present invention provides a pharmaceutical composition for preventing or treating osteoporosis comprising exosomes isolated from bovine colostrum as an active ingredient.

The pharmaceutical composition for preventing or improving osteoporosis according to the present invention contains exosomes isolated from bovine colostrum as an active ingredient.

As used herein, the term "bovine colostrum" refers to breast milk produced from a mother cow during the first about 24-72 hours after the cow's childbirth.

The term "exosomes" herein refers to membrane vesicles (30-150 nm) of intracellular origin, mediating cell-to-cell interactions by the transfer of various proteins, lipids and RNAs between different cell types, and from all biological fluids. It can be separated, characterized in that the exosomes are separated from bovine colostrum.

Specifically, the exosome may inhibit the differentiation of osteoclasts, and may increase the bone density and bone mass percentage of the femur.

On the other hand, if osteoporosis occurs, the permeation cuticle's (firmicutes) or di Perry tumefaciens less intestinal bacteria belonging to the (deferribacteres) can be increased, Lactobacillus bacteria (Lactobacillus) or the thin steroid intestinal microorganisms belonging to the (Bacteroides) can be reduced There, the exosomes can again reduce the intestinal microbes increased by the osteoporosis, and increase the intestinal microbes reduced by the osteoporosis again.

"Osteoporosis" in the present specification is a systemic skeletal disease characterized by low bone mass and microstructural deterioration of bone tissue, postmenopausal osteoporosis, which is caused by increased bone resorption due to activation of osteoclasts, and osteoblast function as aging It can be classified into senile osteoporosis, etc., in which bone formation decreases due to this decrease. Preferably, the osteoporosis may be caused by activation of osteoclasts.

The pharmaceutical composition for preventing or treating osteoporosis according to the present invention is in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. It may be formulated and used, and may include an appropriate carrier, excipient, or diluent commonly used in the preparation of a pharmaceutical composition for formulation.

Examples of the carrier or excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, undecided And various compounds or mixtures including vaginal cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, it can be prepared using diluents or excipients such as commonly used fillers, weight agents, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration may be prepared by mixing at least one excipient, such as starch, calcium bonate, sucrose or lactose, gelatin, and the like with the exosomes. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used.

Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous agents, suspensions, emulsions, lyophilized formulations, and suppositories. As the non-aqueous solvent and suspension, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerol gelatin, and the like can be used.

The preferred dosage of the pharmaceutical composition for preventing or treating osteoporosis according to the present invention varies depending on the patient's condition, weight, degree of disease, drug form, route and duration of administration, but may be appropriately selected by those skilled in the art. However, for a desirable effect, it can be administered at 0.0001 to 2,000 mg/kg per day, preferably at 0.1 mg/kg to 10 mg/kg, and more preferably at 1.0 mg/kg to 10 mg/kg. I can. Administration may be administered once a day, or may be divided several times. However, the scope of the present invention is not limited by the dosage.

The pharmaceutical composition for preventing or treating osteoporosis according to the present invention can be administered to mammals such as mice, mice, livestock, and humans by various routes. All modes of administration can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Health functional food for preventing or improving osteoporosis

In addition, the present invention provides a health functional food for preventing or improving osteoporosis comprising exosomes isolated from bovine colostrum as an active ingredient:

The health functional food for preventing or improving osteoporosis according to the present invention contains exosomes isolated from bovine colostrum as an active ingredient, "exosomes isolated from bovine colostrum (or exosomes derived from bovine colostrum)" and "osteoporosis" Since the above has been described, redundant description will be omitted.

In the health functional food for preventing or improving osteoporosis according to the present invention, when using exosomes (BCE) isolated from bovine colostrum as an additive for a health functional food, it can be added as it is or used with other foods or food ingredients. , It can be appropriately used according to a conventional method. The mixing amount of the active ingredient can be appropriately determined according to each purpose of use, such as prevention, health or treatment.

Formulations of the health functional food may be in the form of powders, granules, pills, tablets, capsules, as well as general foods or beverages.

The type of food is not particularly limited, and examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , Various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and may include all foods in the usual sense.

In general, when preparing food or beverage, exosomes (BCE) separated from bovine colostrum may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on 100 parts by weight of raw materials. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be less than or equal to the above range, and the present invention has no problem in terms of safety in terms of using natural products. Can also be used.

Among the health functional foods according to the present invention, the beverage may contain various flavoring agents or natural carbohydrates as an additional component, like a conventional beverage. The natural carbohydrates described above may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the beverage according to the present invention.

In addition to the above, the health functional food for preventing or improving osteoporosis according to the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers. It may contain an agent, a preservative, a glycerin, an alcohol, a carbonating agent used in carbonated beverages. In addition, the composition for preventing or improving osteoporosis of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These ingredients may be used independently or in combination. The ratio of these additives is not limited, but it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food of the present invention.

Accordingly, the present invention relates to a composition for preventing, treating or improving osteoporosis comprising exosomes isolated from bovine colostrum as an active ingredient, wherein the exosomes can inhibit the differentiation of osteoclasts, and bone density and bone mass percentage of the femur Can increase. In addition, since it can change the intestinal microflora increased or decreased by the osteoporosis, it can be usefully used as a medicine or health functional food for preventing, treating or improving osteoporosis.

According to the present invention, exosomes isolated from bovine colostrum may be potential candidates for preventing osteoporosis, improving bone remodeling, and inhibiting bone resorption. The protective effect of milk exosomes on osteoporosis was first demonstrated in vivo, and exosomes isolated from bovine colostrum can be used as prophylicsis by preventing the onset of osteoporosis. Thus, it can provide a promising alternative to designing new strategies for nutritional management of age-related bone complications.

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention. However, the following examples are provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1: Preparation of bovine colostrum-derived exosomes (BCE)

Using the method described in Maburutse et al., 2017, exosomes were isolated from bovine colostrum (BCE, exosomes derived from bovine colostrum). In summary, the colostrum was sequentially centrifuged to degrease the colostrum, the cell debris was pelleted, and then a whey fraction was prepared, followed by ultracentrifugation at 100,000 g. The supernatant was collected and ultracentrifuged for 90 minutes at 135,000 g to pellet the exosomes. The pellet was then resuspended in PBS, filtered through 0.22 μm and frozen for later use.

Example 2: Confirmation of osteoclast differentiation inhibitory ability of bovine colostrum-derived exosomes (BCE)

The reduction in bone density in bone diseases is caused by the promotion of differentiation and the function of osteoclasts. Thus, reversing the fatal effects of increased osteoclast formation and maturation is important for maintaining normal bone function. RANKL plays a crucial role in inducing osteoclast formation from hematopoietic cells of the monocyte-macrophage lineage, and RANKL-induced differentiation into osteoclasts forms an F-actin loop structure on the bone surface to perform bone resorption. It is characterized by the formation of a pre-stage multinucleated giant cell. To test whether BCE inhibits osteoclast formation, 35 ng/ml of RANKL (R&D system, USA) and 15 ng/ml of macrophage-colony-stimulating factor ( M-CSF, R&D system, USA) were stimulated with RAW 264.7 cells and tested with a diagnostic acidic phosphatase kit (Sigma, Castle Hill, Australia). TRAP positive multinucleated cells (≥ 3 nuclei), considered osteoclasts, were imaged and counted using light microscopy. 1A, RANKL and M-CSF stimulate the differentiation of RAW264.7 into TRAP-positive multinucleated osteoclasts in the vehicle-treated control, whereas the number of osteoclasts was significantly reduced in the presence of BCE. (Figure 1A). In particular, high concentration of BCE (150 ng/ml) strongly inhibited osteoclast formation. These results suggest that BCE can favorably affect bone diseases by inhibiting differentiation into osteoclasts, and that milk-derived exosomes play a role in preventing bone resorption by diet. Interestingly, more than 95% of RAW264.7 cells survived for 4 days in the medium containing BCE.

Example 3: cow colostrum origin Exosomes (BCE) Bone density of the femur and Bone mass Determine whether to increase the percentage

Because milk plays an important role in the growth of newborns, the biologically active components of milk can directly affect bone metabolism. The exosomes present in whey milk have many miRNAs and appear to have various activities that are beneficial to the host. After evaluating the effect of BCE on osteoclast differentiation in vitro, we investigated whether BCE could be beneficial for osteoporosis in an animal model. To study the effect of BCE consumption on the progression of osteoporosis, slow-release pellets (7.5 mg prednisolone, Innovative Research of America, Sarasota, FL, USA) were placed in the shoulders of rats using the method described in Thiele et al, 2014. By placing subcutaneously at the site, osteoporosis was induced through glucocorticoid treatment. The induction of osteoporosis was confirmed by comparing the Sham group who received only surgical treatment without pellets. Mouse (Sham) and osteoporotic (PDS) group mice were fed a normal diet without ingestion of exosomes. Before the pellet is implanted, the mice to be tested are BCE at a dose of 1500 μg/kg/day (PDS + H-BCE)- or 150 μg/kg/day (PDS + L-BCE) by oral gavage for 8 weeks. Received. To investigate the biological effects of BCE on bone health and intestinal microflora, Sham mice fed high concentrations of BCE (H-BCE) were included in animal experiments. All mice were scanned under anesthesia 4 weeks after the pellet was implanted (FIG. 1B ). Osteoporosis is associated with loss of tissue in the cross-sectional area of the bone as well as damage to the microstructure of the bone. In particular, the microstructure and biomechanical properties of the femur provide clear clues explaining the effect of the compound on osteoporosis. 1C and 1D, femoral bone mineral density (BMD) and percent bone mass (PBV) were significantly lower in the glucocorticoid-induced osteoporosis (PDS) group compared to the Sham group (p<0.05). Osteoporotic mice fed a high concentration of colostrum-derived exosomes (PBS + H-BCE) showed higher BMD and PBV compared to osteoporotic mice not fed BCE (PDS). These results indicate that BCE intake significantly (at least partially) prevents glucocorticoid-induced osteoporosis (p <0.01). In micro-CT images of the cancellous bone of the femur for each group (Fig. 1E), even the mice that were chemically induced to have osteoporosis, the BCE-supplied mice showed significantly higher bone density. In the present invention, there was no significant difference between the Sham group and the high concentration BCE-supplied group.

Example 4: Confirmation of whether bovine colostrum-derived exosomes (BCE) change intestinal microflora

Intestinal microbes play an important role in human health. According to a number of studies, it has been reported that intestinal microflora is associated with physiological bone metabolism and bone disease. To test the hypothesis that the anti-osteoporosis activity of BCE is related to rebalancing in the gut microbiota, we analyzed fecal microbial community diversity using UniFrac analysis to compare the degree of microbial phylogenetic similarity (beta diversity) between groups. . Unweighted weighted UniFrac two-dimensional principle coordinate analysis (PCoA) plot shows clusters of moving sites with three groups at the phylum level from PDS group (red circle) to PDS + H-BCE group (green circle) (Fig. 2A). . The alpha diversity of gut microbiota was analyzed based on the diversity index of Chao1 and Shannon; The pattern of these indices tends to recover from the PDS group (red bar) to the PDS + H-BCE group (green bar). Cluster bar plot analysis showed the cluster composition in the three groups, Firmicutes by osteoporosis. And Deferribacteres increased and Bacteroidetes decreased (FIG. 2C). Increased amounts of Firmicutes and Deferribacteres in the PDS group were effectively reduced by BCE. A representative heat map was constructed by selecting the top 30 abundant genera from three groups (Sham, PDS and PDS + H-BCE) (Fig. 2D). Based on the heat map, the relative abundance of Lactobacillus and Bacteroides in the PDS group was relatively decreased compared to that of the Sham group, but the decrease was significantly reversed by BCE uptake (FIGS. 2E and 2F ).

Hereinafter, an example of the preparation of a composition comprising exosomes (BCE) isolated from bovine colostrum of the present invention is described, but the present invention is not intended to limit it, but is intended to be described in detail.

Formulation Example 1: Preparation of powder

20 mg of exosomes (BCE) isolated from bovine colostrum

100 mg lactose hydrate

10 mg of talc

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2: Preparation of tablets

10 mg of exosomes (BCE) isolated from bovine colostrum

100 mg corn starch

100mg lactose hydrate

2mg magnesium stearate

After mixing the above ingredients, tablets were prepared by tableting according to a conventional tablet preparation method.

Formulation Example 3: Preparation of capsules

10 mg of exosomes (BCE) isolated from bovine colostrum

Microcrystalline cellulose 3 mg

Lactose Hydrate 14.8 mg

Magnesium stearate 0.2 mg

After mixing the above ingredients, the capsules were prepared by filling them into gelatin capsules according to a conventional capsule preparation method.

Formulation Example 4: Preparation of injection

10 mg of exosomes (BCE) isolated from bovine colostrum

180mg mannitol

2974 mg of sterile distilled water for injection

Sodium monohydrogen phosphate 26 mg

After mixing the above ingredients, it was prepared in the amount of the above ingredients per ampoule (2 mL) according to a conventional method for preparing injections.

Formulation Example 5: Preparation of liquid formulation

10 mg of exosomes (BCE) isolated from bovine colostrum

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

Lemon flavor appropriate amount

The above ingredients are dissolved by adding each ingredient to purified water according to a conventional manufacturing method, added an appropriate amount of lemon flavor, adjusted to 100 mL by adding purified water, sterilized, and filled in a brown bottle to prepare a liquid formulation.

Formulation Example 6: Preparation of health functional food

10 mg of exosomes (BCE) isolated from bovine colostrum

The right amount of vitamin mixture

Vitamin A acetate 70 ㎍

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

Vitamin B ₁₂ 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

Nicotinic acid amide 1.7 mg

Folic acid 50 ㎍

Calcium pantothenate 0.5 mg

Mineral mixture Appropriate amount

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium Phosphate Monobasic 15 mg

Dicalcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for the health functional food, but it is also possible to arbitrarily modify the composition, and the above ingredients are mixed according to the general health functional food manufacturing method. Then, granules are prepared, and can be used in the manufacture of health functional foods according to a conventional method.

Formulation Example 7: Preparation of health drink

10 mg of exosomes (BCE) isolated from bovine colostrum

Vitamin C 15 g

100 g of vitamin E (powder)

19.75 g of iron lactate

Zinc oxide 3.5 g

3.5 g of nicotinic acid amide

0.2 g of vitamin A

0.25 g of vitamin B ₁

0.3 g of vitamin B ₂

Purified water quantification

After mixing the above ingredients according to a conventional health drink manufacturing method, the resulting solution is stirred and heated at 85°C for about 1 hour, and the resulting solution is filtered and obtained in a sterilized 2 liter container, sealed and sterilized, and then stored in a refrigerator. It is used in the manufacture of health drink composition.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the purpose of use.

The above description of the present invention is for illustrative purposes only, and those of ordinary skill in the art to which the present invention pertains will be able to understand that other specific forms can be easily modified without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and are not limiting.

Claims (12)

A pharmaceutical composition for preventing or treating osteoporosis comprising exosomes isolated from bovine colostrum as an active ingredient.
The method of claim 1,
The exosome is characterized in that to inhibit the differentiation of osteoclasts, osteoporosis prevention or treatment pharmaceutical composition.
The method of claim 1,
The exosome is characterized in that to increase the bone density and bone mass percentage of the femur, osteoporosis prevention or treatment pharmaceutical composition.
The method of claim 1,
The exosome is characterized in that to again reduce the intestinal microbes increased by the osteoporosis, and again increase the intestinal microbes reduced by the osteoporosis, a pharmaceutical composition for preventing or treating osteoporosis.
The method of claim 4,
Intestinal microflora of the increased permeation cuticle's (firmicutes) or di Perry tumefaciens less belongs to (deferribacteres), wherein the reduce intestinal microorganisms are Lactobacillus bacteria (Lactobacillus) or night, prevention of osteoporosis, characterized in that belonging to the steroid (Bacteroides) Or a pharmaceutical composition for treatment.
The method of claim 1,
The daily dosage of the composition is 0.1 mg/kg to 10 mg/kg, characterized in that, a pharmaceutical composition for preventing or treating osteoporosis.
Health functional food for preventing or improving osteoporosis containing exosomes isolated from bovine colostrum as an active ingredient.
The method of claim 7,
The exosome is a health functional food for preventing or improving osteoporosis, characterized in that it inhibits the differentiation of osteoclasts.
The method of claim 7,
The exosome is a health functional food for preventing or improving osteoporosis, characterized in that to increase the bone density and bone mass percentage of the femur.
The method of claim 7,
The exosome is characterized in that to again reduce the intestinal microorganisms increased by the osteoporosis, and again increase the intestinal microorganisms reduced by the osteoporosis, osteoporosis prevention or improvement health functional food.
The method of claim 10,
Intestinal microflora of the increased permeation cuticle's (firmicutes) or di Perry tumefaciens less belongs to (deferribacteres), wherein the reduce intestinal microorganisms are Lactobacillus bacteria (Lactobacillus) or night, prevention of osteoporosis, characterized in that belonging to the steroid (Bacteroides) Or health functional food for improvement.
The method of claim 7,
The daily dosage of the composition is 0.1 mg/kg to 10 mg/kg, characterized in that the health functional food for preventing or improving osteoporosis.

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