Food composition for preventing or alleviating osteoporosis, containing propolis bioconversion extract as active component

Abstract

The present invention relates to a food composition for preventing or alleviating osteoporosis, the food composition containing a propolis bioconversion extract as an active component. The present invention can provide a food composition for preventing or alleviating osteoporosis, wherein the food composition can be easily and simply prepared and has an excellent osteoporosis prevention effect. In addition, the present invention can provide a food composition for preventing or alleviating osteoporosis, wherein side effects such as weight gain or headaches due to long-term ingestion can be minimized.

Description

Food composition for preventing or improving osteoporosis containing propolis bioconversion extract as an active ingredient

[0001] The present invention relates to a food composition for preventing or improving osteoporosis, comprising a propolis bioconversion extract as an active ingredient.

[0002]

[0003] Various plant-derived chemical substances are attracting attention as useful materials for health functional foods because they exhibit various physiological activities, including anti-osteoporosis effects, inhibition of osteoclast differentiation, and cytoprotection.

[0004] Meanwhile, osteoporosis, which is increasing not only in Korea but also worldwide due to aging, lack of exercise, and smoking, is a disease that causes serious risks such as multiple fractures and hip fractures, and significantly reduces the quality of life in old age. Therefore, the development of bioactive substances to prevent and treat osteoporosis is required.

[0005] In this regard, Korean Patent Publication No. 10-2004-0088976 discloses a composition for preventing and improving osteoporosis, comprising (a) an alkaline aqueous solution for mineral supplementation comprising beef bones, squid bones, red algae, organic acid, and purified water; (b) a herbal composition; (c) dried pollack; (d) black beans; (e) shiitake mushrooms; (f) casein phosphopeptide; and (g) DHA (Docosahexaenoic acid).

[0006] However, the technology disclosed in the above document utilizes an alkaline aqueous solution, so the manufacturing method is complex and the efficacy is not excellent.

[0007] Additionally, there is a problem that long-term use can cause side effects such as weight gain, headaches, and depression.

[0008]

[0009] (Patent Document 0001) Korean Patent Publication No. 10-2004-0088976

[0010]

[0011] The present invention is intended to solve the problems of the above-mentioned prior art, and aims to provide a food composition for preventing or improving osteoporosis that can be easily and simply manufactured and has an excellent osteoporosis prevention effect.

[0012] In addition, the present invention aims to provide a food composition for preventing or improving osteoporosis that can minimize side effects such as weight gain and headache when consumed for a long period of time.

[0013]

[0014] In order to achieve the above purpose, the present invention provides a food composition for preventing or improving osteoporosis, which contains a propolis bioconversion extract as an active ingredient.

[0015] In one embodiment of the present invention, the propolis bioconversion extract is prepared by mixing propolis and alcohol, extracting by heating, and then concentrating under reduced pressure;

[0016] A step of producing a fermented product by inoculating the above extract with Lactifrantibacillus plantarum; and

[0017] It is characterized by being manufactured through a step of adding ethyl acetate to the above fermented product, centrifuging, and then concentrating the ethyl acetate layer under reduced pressure to obtain a solid extract.

[0018] In one embodiment of the present invention, the propolis is characterized in that it is Australian propolis.

[0019] In one embodiment of the present invention, the composition is characterized in that it further comprises a propolis extract.

[0020] In one embodiment of the present invention, the weight ratio of the propolis bioconversion extract and the propolis extract is characterized by being 70 to 90:10 to 30.

[0021]

[0022] The present invention can provide a food composition for preventing or improving osteoporosis that can be easily and simply manufactured and has an excellent osteoporosis prevention effect.

[0023] In addition, the present invention can provide a food composition for preventing or improving osteoporosis that can minimize side effects such as weight gain and headache when consumed for a long period of time.

[0024]

[0025] Figure 1 shows the TRAP activity of the propolis bioconversion extract of the present invention.

[0026] Figure 2 shows the TRAP staining characteristics of the propolis bioconversion extract of the present invention: A control, B untreated sample, C propolis extract treated sample, D propolis bioconversion extract treated sample.

[0027]

[0028] The present invention will be described in detail below based on examples. Terms, examples, and the like used herein are merely illustrative examples to more specifically explain the invention and aid understanding by those skilled in the art. The scope of the invention should not be construed as being limited thereto.

[0029] Technical and scientific terms used in the present invention, unless otherwise defined, have the meaning commonly understood by a person of ordinary skill in the art to which this invention belongs.

[0030]

[0031] The present invention relates to a food composition for preventing or improving osteoporosis, comprising a propolis bioconversion extract as an active ingredient.

[0032]

[0033] The propolis bioconversion extract of the present invention can be used to prevent or improve osteoporosis.

[0034] The above propolis bioconversion extract exhibits high peroxyl radical scavenging activity and reducing capacity, effectively reduces intracellular oxidative stress, exhibits excellent intracellular antioxidant activity, and has high osteoclast differentiation inhibition activity, making it effective in preventing and improving osteoporosis.

[0035] Bioconversion refers to a technology that converts existing materials by utilizing the functions of living organisms, including microorganisms, or biocatalysts such as enzymes.

[0036] The present invention can produce a propolis bioconversion extract by bioconverting a propolis extract using an enzyme produced by a strain.

[0037]

[0038] The above propolis bioconversion extract is prepared by mixing propolis and alcohol, heating and extracting them, and then concentrating under reduced pressure;

[0039] A step of producing a fermented product by inoculating the above extract with Lactifrantibacillus plantarum; and

[0040] It can be manufactured through a step of adding ethyl acetate to the above fermented product, centrifuging, and then concentrating the ethyl acetate layer under reduced pressure to obtain a solid extract.

[0041]

[0042] The step of manufacturing the above extract can be performed by mixing propolis with alcohol such as ethanol, methanol, alcohol, or fermented alcohol, heating the mixture to extract it, and then concentrating it under reduced pressure to manufacture the extract.

[0043] The above heat extraction can be performed by adding 100 to 5,000 parts by weight of alcohol to 100 parts by weight of propolis and heating at 40 to 90°C for 1 to 20 hours.

[0044] Propolis extract can be obtained by heating, extracting, and then concentrating under reduced pressure.

[0045] At this time, it is preferable that the propolis is Australian propolis.

[0046]

[0047] The step of manufacturing the above fermented product can be performed by inoculating the above extract with Lactiplantibacillus plantarum and fermenting it to manufacture a propolis fermented product.

[0048] After mixing the above extract, ethanol and polyethylene glycol, 0.5 to 10% of Lactiplantibacillus plantarum can be inoculated and fermentation can be performed at 10 to 60°C for 5 to 100 hours.

[0049]

[0050] The step of obtaining the above solid extract can be performed by adding ethyl acetate to the above fermented product, centrifuging, and then concentrating the ethyl acetate layer under reduced pressure to obtain the solid extract.

[0051] Ethyl acetate can be added to the above fermented product, allowed to stand at room temperature for 1 to 100 minutes, and then centrifuged for 1 to 30 minutes to remove solid residue.

[0052] After that, the ethyl acetate layer can be concentrated under reduced pressure to obtain a solid extract.

[0053]

[0054] The present invention also relates to a propolis bioconversion extract and a food composition for preventing or improving osteoporosis comprising a propolis extract as an active ingredient.

[0055] At this time, the weight ratio of the propolis bioconversion extract and the propolis extract is preferably 70 to 90:10 to 30, and when the weight ratio satisfies the above numerical range, the osteoporosis prevention effect can be maximized.

[0056]

[0057] The above propolis extract can be prepared by mixing propolis with alcohol such as ethanol, methanol, alcohol, or fermented alcohol, heating the mixture to extract it, and then concentrating it under reduced pressure.

[0058] The above heat extraction can be performed by adding 100 to 5,000 parts by weight of alcohol to 100 parts by weight of propolis and heating at 40 to 90°C for 1 to 20 hours.

[0059] Propolis extract can be obtained by heating, extracting, and then concentrating under reduced pressure.

[0060]

[0061] The present invention also relates to a food composition for preventing or improving osteoporosis, comprising a propolis bioconversion extract and a propolis additional bioconversion extract as active ingredients.

[0062] At this time, the weight ratio of the propolis bioconversion extract and the propolis additional bioconversion extract is preferably 70 to 90:10 to 30, and when the weight ratio satisfies the above numerical range, the osteoporosis prevention effect can be maximized.

[0063]

[0064] The above propolis additional bioconversion extract is a step of producing a first fermented product by inoculating propolis with Lactifrantibacillus plantarum;

[0065] A step of preparing an extract by mixing the first fermentation product and alcohol, heating and extracting, and then concentrating under reduced pressure;

[0066] A step of producing a second fermented product by inoculating the above extract with Lactifrantibacillus plantarum; and

[0067] It can be manufactured through a step of adding ethyl acetate to the second fermentation product, centrifuging, and then concentrating the ethyl acetate layer under reduced pressure to obtain a solid extract.

[0068] The step of manufacturing the above first fermented product can be performed by mixing propolis, ethanol, and polyethylene glycol, inoculating 0.5 to 10% of Lactiplantibacillus plantarum, and fermenting at 10 to 60°C for 5 to 100 hours.

[0069] After the fermentation process is completed, the first fermented product can be obtained by concentrating under reduced pressure.

[0070] Additionally, the step of preparing an extract, the step of preparing a second fermentation product, and the step of obtaining a solid extract can be performed in the same manner as the process for preparing a propolis bioconversion extract.

[0071]

[0072] The present invention also relates to a food composition for preventing or improving osteoporosis, comprising a propolis bioconversion extract, a propolis extract, and a propolis additional bioconversion extract as active ingredients.

[0073] At this time, the weight ratio of the propolis bioconversion extract, propolis extract, and propolis additional bioconversion extract is preferably 100:5 to 15:10 to 30, and when the weight ratio satisfies the above numerical range, the osteoporosis prevention effect can be maximized.

[0074]

[0075] The food of the present invention may be in the form of various foods, candies, beverages, gums, teas, vitamin complexes, health functional foods, etc., and may be provided in the form of powders, granules, tablets, capsules, beverages, etc.

[0076]

[0077] The present invention also relates to a pharmaceutical composition for preventing or treating osteoporosis, comprising a propolis bioconversion extract as an active ingredient.

[0078] The above pharmaceutical composition may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

[0079] The carriers, excipients and diluents may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0080] The pharmaceutical composition of the present invention can be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions.

[0081]

[0082] The present invention is described in detail through the following examples. The following examples are merely illustrative for the implementation of the present invention, and the scope of the present invention is not limited to the following examples.

[0083]

[0084] (Example 1) Preparation of propolis extract

[0085] A propolis extract was prepared by mixing 100 parts by weight of Australian propolis and 500 parts by weight of 95% ethanol, heating and extracting at 50°C for 10 hours, and then concentrating under reduced pressure.

[0086]

[0087] (Example 2) Preparation of propolis bioconversion extract

[0088] An extract was prepared by mixing 100 parts by weight of Australian propolis and 500 parts by weight of 95% ethanol, heating and extracting at 50°C for 10 hours, and then concentrating under reduced pressure.

[0089] After mixing the above extract, ethanol and polyethylene glycol, 1.5% of Lactiplantibacillus plantarum (owned by Megalab Co., Ltd.) derived from kimchi was inoculated and fermented at 30°C for 48 hours to produce a fermented product.

[0090] Ethyl acetate was added to the above fermented product, allowed to stand at room temperature for 10 minutes, and then centrifuged for 5 minutes to remove solid residue.

[0091] Afterwards, the ethyl acetate layer was concentrated under reduced pressure to prepare a propolis bioconversion extract.

[0092]

[0093] (Example 3) Preparation of propolis additional bioconversion extract

[0094] After mixing Australian propolis, ethanol, and polyethylene glycol, 1.5% of Lactiplantibacillus plantarum (owned by Megalab Co., Ltd., derived from kimchi) was inoculated and fermented at 30°C for 48 hours. After the fermentation process was completed, the first fermented product was produced by concentration under reduced pressure.

[0095] After mixing 100 parts by weight of the first fermentation product and 500 parts by weight of 95% ethanol, the mixture was heated and extracted at 50°C for 10 hours and then concentrated under reduced pressure to prepare an extract.

[0096] After mixing the above extract, ethanol and polyethylene glycol, 1.5% of Lactiplantibacillus plantarum (owned by Megalab Co., Ltd.) derived from kimchi was inoculated and fermented at 30°C for 48 hours to produce a second fermented product.

[0097] Ethyl acetate was added to the above second fermentation product, allowed to stand at room temperature for 10 minutes, and then centrifuged for 5 minutes to remove solid residue.

[0098] Afterwards, the ethyl acetate layer was concentrated under reduced pressure to prepare a propolis additional bioconversion extract.

[0099]

[0100] (Example 4) Total polyphenol content

[0101] To determine the total polyphenol content, 5 ml of the extract sample was collected and diluted with 5 ml of 10% ethanol. A 0.25 ml portion of each sample was then added to 95% ethanol containing 1% HCl and 4.55 ml of 2% HCl. After reacting for 15 minutes, the absorbance value of each sample was measured at 280 nm using a spectrophotometer.

[0102] Total polyphenol content was determined using a curve for gallic acid as a standard.

[0103] The total polyphenol content of the propolis extract of Example 1 was 0.48 mM / L, and the total polyphenol content of the propolis bioconversion extract of Example 2 was 0.52 mM / L, confirming that the polyphenol content increased through fermentation.

[0104]

[0105] (Example 5) TRAP activity of propolis extract

[0106] Tartrate resistant acid phosphatase (TRAP) activity was measured by seeding 1 × 10 RAW 264.7 cells in a 96-well plate. 4 The cells were pre-cultured for 24 hours at a number of cells / well, and the samples (propolis extract) were treated at various concentrations with differentiation-inducing medium [50 ng / mL receptor activator of nuclear factor-κB ligand (RANKL)].

[0107] Samples were added every two days when the medium was changed and cultured for a total of five days.

[0108] After removing the media, the cells were washed once with 50 μl of PBS (pH 7.4), and then 100 μl of 50 mM citrate buffer containing 10 mM sodium tartrate and 5 mM ρ-nitrophenylphosphate was dispensed, followed by reaction at 37°C for 1 hour.

[0109] After the reaction, the reaction was stopped with 0.1 N NaOH and the absorbance was measured at 405 nm.

[0110]

[0111] The results of measuring the activity of tartrate resistant acid phosphatase (TRAP), which is expressed specifically in osteoclasts, after differentiation of RAW 264.7 cells for 5 days by RANKL, an osteoclast differentiation inducer, using propolis extract are presented in Figure 1.

[0112] When the TRAP activity of the untreated sample (only RANKL treatment) was taken as 100%, the TRAP activity was 62.8% when 0.005% of the propolis extract of Example 1 was treated, and the TRAP activity was 49.8% when 0.005% of the propolis bioconversion extract of Example 2 was treated.

[0113] Propolis bioconversion extract, which is produced by fermenting or bioconverting propolis extract, can effectively inhibit the activity of TRAP induced by RANKL.

[0114] These results suggest that propolis bioconversion extract may have a positive effect on bone health by effectively inhibiting osteoclast formation and TRAP activity, thereby hindering bone resorption.

[0115]

[0116] (Example 6) TRAP staining of propolis extract

[0117] RAW 264.7 cells were seeded at 1×10 in a 24-well plate. 5After 24 hours of pre-incubation with the number of cells / well, the sample (propolis extract) was treated with differentiation-inducing medium [50 ng / mL receptor activator of nuclear factor-κB ligand (RANKL)] at various concentrations.

[0118] Samples were added every two days when the medium was changed and cultured for a total of five days.

[0119] After removing the media, the cells were washed once with 500 μl of PBS (pH 7.4), fixed with 4% formaldehyde for 10 minutes, and then fixed with ethanol / acetone (1:1) for 1 minute.

[0120] After washing twice with distilled water, TRAP staining was performed using leukocyte acid phosphatase kit 387-A (Sigma).

[0121] After staining, TRAP-positive multinucleated cells with three or more nuclei were counted using an optical microscope and used as an indicator of osteoclast production.

[0122]

[0123] The results of TRAP staining of propolis extract samples are presented in Figure 2.

[0124] As a result of identifying TRAP-positive multinucleated osteoclasts, propolis bioconversion extract (D) was found to reduce osteoclast formation at a concentration of 0.005%.

[0125]

[0126] (Example 7) TRAP activity of propolis mixed extract

[0127] Table 1 below shows the TRAP activity according to the mixture of propolis extracts.

[0128]

[0129] TRAP activity (%) RANKL only treatment 100 Propolis extract of Example 1 62.8 Propolis bioconversion extract of Example 2 49.8 Example 1 + Example 2 (20:80) 45.3 Example 1 + Example 2 (40:60) 60.4 Example 1 + Example 2 (5:95) 58.6 Example 3 + Example 2 (20:80) 43.9 Example 3 + Example 2 (40:60) 55.7 Example 3 + Example 2 (5:95) 57.1 Example 1 + Example 2 + Example 3 (10:100:20) 40.8 Example 1 + Example 2 + Example 3 (10:100:5) 56.6 Example 1 + Example 2 + Example 3 (10:100:35) 59.5

[0130]

[0131] As can be seen from the above table, the propolis bioconversion extract and the propolis extract can be used in combination, and when the weight ratio of the propolis bioconversion extract and the propolis extract is 70~90:10~30, the osteoporosis prevention effect is maximized.

[0132] In addition, the propolis bioconversion extract and the propolis additional bioconversion extract can be used in combination, and it can be seen that the osteoporosis prevention effect is maximized when the weight ratio of the propolis bioconversion extract and the propolis additional bioconversion extract is 70~90:10~30.

[0133] In addition, propolis bioconversion extract, propolis extract, and propolis additional bioconversion extract can be used in combination, and when the weight ratio of propolis bioconversion extract, propolis extract, and propolis additional bioconversion extract is 100:5~15:10~30, the osteoporosis prevention effect can be maximized.

[0134]

[0135] The present invention can provide a food composition for preventing or improving osteoporosis that can be easily and simply manufactured and has an excellent osteoporosis prevention effect.

[0136] In addition, the present invention can provide a food composition for preventing or improving osteoporosis that can minimize side effects such as weight gain and headache when consumed for a long period of time.

Claims

1. A food composition for preventing or improving osteoporosis, comprising a propolis bioconversion extract as an active ingredient.

2. In paragraph 1, The above propolis bioconversion extract is A step of preparing an extract by mixing propolis and alcohol, extracting by heating, and then concentrating under reduced pressure; A step of producing a fermented product by inoculating the above extract with Lactifrantibacillus plantarum; and A food composition for preventing or improving osteoporosis, characterized in that it is manufactured through a step of adding ethyl acetate to the above fermented product, centrifuging the product, and then concentrating the ethyl acetate layer under reduced pressure to obtain a solid extract.

3. In paragraph 2, A food composition for preventing or improving osteoporosis, characterized in that the above propolis is Australian propolis.

4. In paragraph 1, A food composition for preventing or improving osteoporosis, characterized in that it further comprises a propolis extract.

5. In paragraph 4, A food composition for preventing or improving osteoporosis, characterized in that the weight ratio of the propolis bioconversion extract and the propolis extract is 70 to 90:10 to 30.

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