ANTI-INFLUENZA VIRUS COMPOSITION, COMPOSITION FOR TREATING RESPIRATORY DISEASES, AND ANTI-AGING COMPOSITION, COMPRISING DARK GINSENG EXTRACT

Abstract

The present invention relates to an anti-influenza virus composition, a composition for treating respiratory diseases, and an anti-aging composition, each of which comprises a dark ginseng extract.

Claims

1. A method for preparing a black ginseng extract enriched with ginsenosides Rk1 and Rg5, the method comprising: a step for steaming ginseng to prepare black ginseng; a step for extracting the prepared black ginseng with a solvent; and a step for maturing the obtained black ginseng extract.

2. The method for preparing a black ginseng extract of claim 1, wherein the steaming is performed at 70° C. to 120° C. for 3 to 12 times.

3. The method for preparing a black ginseng extract of claim 1, wherein the steaming is performed under the condition of 2 hours or more per steaming.

4. The method for preparing a black ginseng extract of claim 1, wherein the step for maturing the black ginseng extract is performed at 80° C. or higher.

5. The method for preparing a black ginseng extract of claim 1, wherein the step for maturing the black ginseng extract is performed for 3 hours or more.

6. The method for preparing a black ginseng extract of claim 1, wherein the total content of ginsenosides Rk1 and Rg5 contained in the black ginseng extract is 20 parts by weight to 90 parts by weight, based on 100 parts by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5, and Rh1(s).

7. A composition for preventing, inhibiting, or treating a disease caused by an influenza virus, the composition comprising a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is 20 parts by weight to 90 parts by weight, based on 100 parts by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5, and Rh1(s).

8. The composition for preventing, inhibiting, or treating a disease caused by an influenza virus of claim 7, wherein the black ginseng extract further comprises acidic polysaccharides and polyphenols.

9. The composition for preventing, inhibiting, or treating a disease caused by an influenza virus of claim 7, wherein the influenza virus is an influenza A virus.

10. The composition for preventing, inhibiting, or treating a disease caused by an influenza virus of claim 9, wherein the influenza virus is H1N1.

11. The composition for preventing, inhibiting, or treating a disease caused by an influenza virus of claim 7, wherein the composition is a food or pharmaceutical composition.

12. The composition for preventing, inhibiting, or treating a disease caused by an influenza virus of claim 7, wherein the disease is any one or more selected from the group consisting of cold, flu, cough, sneezing, runny nose, myalgia, pharyngolaryngitis, nasal obstruction, laryngitis, sore throat, hoarseness, headache, sinus pain, rhinitis, pharyngitis, bronchitis, asthma, fever, dyspnea, whole body lethargy, and chills.

13. A composition for preventing, inhibiting, or treating a respiratory disease, the composition comprising a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is 20 parts by weight to 90 parts by weight, based on 100 parts by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3 (s), Rk1, Rg5, and Rh1(s).

14. The composition for preventing, inhibiting, or treating a respiratory disease of claim 13, wherein the black ginseng extract further comprises acidic polysaccharides and polyphenols.

15. The composition for preventing, inhibiting, or treating a respiratory disease of claim 13, wherein the respiratory disease is selected from the group consisting of asthma, chronic obstructive pulmonary disease, bronchitis, pharyngitis, laryngitis, rhinitis, sinusitis, and pneumonia.

16. The composition for preventing, inhibiting, or treating a respiratory disease of claim 15, wherein the respiratory disease is caused by fine dust or ultrafine dust.

17. An anti-aging composition comprising a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is 20 parts by weight to 90 parts by weight, based on 100 parts by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5, and Rh1(s).

18. The anti-aging composition of claim 17, wherein the black ginseng extract further comprises acidic polysaccharides and polyphenols.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0117] FIGS. 1a to 1c show changes in ginsenosides according to the steaming time during the preparation of the black ginseng.

[0118] FIGS. 2a to 2c show changes in ginsenosides according to the maturing temperature and the maturing time of the black ginseng concentrate.

[0119] FIG. 3 is a graph showing the fatality rate after the virus infection of the standardized black ginseng concentrate treatment group.

[0120] FIG. 4 shows the degree of lung tissue damage after the virus infection of the standardized black ginseng concentrate treatment group.

[0121] FIGS. 5a to 5c are graphs showing the degree of immune activation factor production after the virus infection of the standardized black ginseng concentrate treatment group.

BEST MODE FOR CARRYING OUT THE INVENTION

[0122] Hereinafter, the present invention will be described in detail through Preparation Examples and Experimental examples.

[0123] However, the following Preparation Examples and Experimental examples merely illustrate the present invention, and the contents of the present application are not limited by the following Examples and Experimental examples.

Preparation Example 1. Preparation of Black Ginseng

[0124] 1-1. Preparation of Red Ginseng (One Steaming-Drying Black Ginseng)

[0125] Four-year-old fresh ginseng roots were placed into a screw washer (Samgak fmc, South Korea) and washed once for 3 minutes to remove soil or foreign substances. The washed fresh ginseng roots were placed in a steamer and first steamed at 98° C. for 2 hours, excluding the preheating time. The steamer was cooled to a temperature of 55° C. Then, the steamed fresh ginseng roots were taken out of the steamer. The internal temperature of a hot air dryer was maintained at 55° C., and then, the steamed fresh ginseng roots were added and dried for about 18 hours. The dried red ginseng was moved to a sun-drying place (a drying facility under natural light) and dried for over a month. Through the above process, 930 kg of red ginseng (one steaming-drying) was prepared.

[0126] 1-2. Preparation of Nine Times Steaming-Nine Times Drying Black Ginseng

[0127] 930 kg of red ginseng prepared in Preparation Example 1-1 above was steamed under the same steaming and drying conditions as in Preparation Example 1-1. The above-described steaming and drying processes were repeatedly performed and were referred to as two times to nine times steaming-drying depending on the number of repetitions. As the steaming and drying were repeated, the color gradually became closer to black. Through the above process, 853.8 kg of nine times steaming-nine times drying black ginseng was prepared. When preparing the black ginseng extract of Preparation Example 2 or the black ginseng concentrate of Preparation Example 3, nine times steaming-nine times drying black ginseng was used as a raw material.

Preparation Example 2. Preparation of Black Ginseng Extract

[0128] Each of 284.6 kg of the red ginseng prepared in Preparation Example 1-1 and 284.6 kg of the black ginseng prepared in Preparation Example 1-2 was taken and pulverized. Then, 70% (v/v) aqueous ethanol solution corresponding to 6 times the weight of the pulverized product was added, and the first extraction was performed for 6 hours at 82° C. The extraction residue generated from the first extraction was subjected to the second extraction under the same conditions as in the first extraction. 50% (v/v) aqueous ethanol solution corresponding to 6 times the weight of the extraction residue generated from the second extraction was added, and the third extraction was performed for 6 hours at 85° C. 30% (v/v) aqueous ethanol solution corresponding to 6 times the weight of the extraction residue generated from the third extraction was added, and the fourth extraction was performed for 6 hours at 90° C. Purified water corresponding to 6 times the weight of the extraction residue generated from the fourth extraction was added, and the fifth extraction was performed for 6 hours at 95° C.

Preparation Example 3. Preparation of Black Ginseng Concentrate

[0129] The first to fourth red ginseng extracts and the first to fourth black ginseng extracts obtained in Preparation Example 2 each were filtered to separate the extracts from the extraction residues. For each raw material, only the separated extract was placed in a concentrator and concentrated at 55° C. in a vacuum concentrator for 4 hours. 231 kg of 70 Brix % red ginseng concentrate was obtained, and 212 kg of 70 Brix % black ginseng concentrate was obtained. The obtained red ginseng concentrate and black ginseng concentrate were used as samples for the following component analysis and influenza A virus experiment.

Experimental Example 1. Confirmation of Changes in Ginsenosides Content in Black Ginseng Concentrate According to Steaming Time

[0130] Nine times steaming-drying was carried out in the same manner as in the steaming conditions of Preparation Example 1-1 except that the steaming time was changed to 1 hour, 2 hours, and 3 hours. The prepared red ginseng was extracted and concentrated according to Preparation Examples 2 and 3. The ginsenosides content was measured in the same manner as in Experimental Example 3 below.

[0131] When the steaming time was set to 1 hour, it was confirmed that ginsenosides Rk1 and Rg5 did not exist even after nine times steaming-drying as shown in FIG. 1a. Meanwhile, when the steaming time was set to 2 hours, the content of ginsenosides Rk1 and Rg5 was significantly increased compared to the black ginseng concentrate prepared with the steaming time of 1 hour (FIG. 1b). In addition, when the steaming time was set to 3 hours, the content of ginsenosides Rk1 and Rg5 was increased compared to the case where the steaming time was set to 2 hours (FIG. 1c).

Experimental Example 2. Confirmation of Changes in Ginsenosides Content in Black Ginseng Concentrate According to Maturing Temperature or Maturing Time

[0132] The black ginseng concentrate obtained in Preparation Example 3 was placed in a 2 ml vial and matured in a water bath for 8 hours at 70° C., 80° C., and 90° C., respectively, and the changes in ginsenosides content over time were measured. The ginsenosides content was measured in the same manner as in Experimental Example 3.

[0133] As a result, when maturing the black ginseng concentrate at 70° C., the ginsenosides content did not increase even after 8 hours of maturing (FIG. 2a). Meanwhile, when maturing the black ginseng concentrate at 70° C., the content of ginsenosides Rk1 and Rg5 increased over time. Even at 90° C., the content of ginsenosides Rk1 and Rg5 increased over time as in the case of maturing at 80° C.

[0134] In addition, as can be seen from FIGS. 2b to 2c, the content of ginsenosides Rk1 and Rg5 was changed according to the maturing time. The content of ginsenosides Rk1 and Rg5 increased in the black ginseng concentrate matured under the condition of 80° C. or higher for 3 hours or more.

Experimental Example 3. Analysis of Ginsenoside Components in Black Ginseng Concentrate

[0135] The analysis of ginsenoside components present in the red ginseng concentrate or the black ginseng concentrate obtained in Preparation Example 3 was performed using HPLC. As a sample, a solution obtained by diluting the black ginseng concentrate obtained in Preparation Example 3 in methanol at a ratio of 1/30, followed by filtration through a 0.45 μm filter, was used. HPLC 1260 (DAD) (Agilent, USA) was used. VENUSIL XBP C18 column (4.6 MM×250 MM, 5.0 μm) was used, and as mobile phases, LC-grade water was used as solvent A and acetonitrile as solvent B. The column temperature was maintained at 35° C. The gradient conditions were as follows: 0-3.8 min, a flow rate of 1 ml/min, hold at 30% B; 3.81-4.5 min, a flow rate of 0.5 ml/min, hold at 30% B; 4.51-6 min, a flow rate of 1.2 ml/min, 30% B increased to 42% B; 7.5-13 min, a flow rate of 0.5 ml/min, hold at 42% B; until 18 min, a flow rate of 0.5 ml/min, increased to 47.4% B; 18.1-25 min, a flow rate of 1 ml/min, increased to 55% B; 30-32 min, a flow rate of 1 ml/min, increased to 60% B; 42-50 min, a flow rate of 1 ml/min, 60% B increased to 90% B; and 50.1-53 min, a flow rate of 1 ml/min, hold at 30% B. The results of the component analysis are shown below (unit: mg/g).

TABLE-US-00001 TABLE 1 Ginsenoside content Rg3 Rh1 Rb1 Rb2 Rc Rd Re Rg1 (S) Rk1 Rg5 (S) Red 4.91 2.21 3.23 1.75 3.74 1.29 0.42 0.12 0.46 0.43 ginseng Black 0.83 0.00 0.00 0.00 1.34 0.00 4.12 4.75 4.54 0.94 ginseng

[0136] As a result of analyzing ginsenosides in the black ginseng concentrate, it was confirmed that 0.832 mg/g of Rb1, 1.157 mg/g of Rf, 1.008 mg/g of Rg2(S), 0.265 mg/g of Rg2(R), 0.941 mg/g of Rh1(S), 0.416 mg/g of Rh1(R), 0.274 mg/g of Rg6, 0.777 mg/g of F4, 0.862 mg/g of Rk3, 1.714 mg/g of Rh4, 4.122 mg/g of Rg3(S), 1.184 mg/g of Rg3(R), 4.747 mg/g of Rk1, and 4.539 mg/g of Rg5 existed.

[0137] When comparing the ginsenosides content contained in the black ginseng concentrate and the red ginseng concentrate, Rk1 and Rg5 increased about 40 times and about 10 times, respectively, compared to those in the red ginseng concentrate.

Experimental Example 4. Analysis of Acidic Polysaccharide Components in Black Ginseng Concentrate

[0138] The analysis of acidic polysaccharide components present in the red ginseng concentrate or the black ginseng concentrate obtained in Preparation Example 3 was performed through the carbazole-sulfuric acid colorimetric method. A sample obtained by diluting 300 mg of each concentrate obtained in Preparation Example 3 above in 10 ml of distilled water was used. The diluted solution was heated in boiling water at 90° C. for 3 hours, cooled, and centrifuged (3000 rpm, 10 minutes). 1 ml of the supernatant was taken, and 4 ml of ethanol was added to form a white precipitate. To obtain the white precipitate, the supernatant was removed after centrifugation (3000 rpm, 10 minutes), 4 ml of distilled water was added to dissolve the white precipitate, and 1 ml of 1:4 mixture of n-butanol and CHCl.sub.3 was added, stirred, and then centrifuged again (3000 rpm, 10 minutes). 4 ml of water extract, which is a supernatant herein, was taken, and sonication was performed.

[0139] 20 μl of the sonication-completed solution, 80 μl of distilled water, 50 μl of 0.1% carbazole ethanol reagent (prepared with 0.125 g of carbazole/100 ml of anhydrous ethanol), and 600 μl of sulfuric acid, were added to a 2 ml tube and stirred. Then, 200 μl each was placed in a 96-well plate. The absorbance was measured at 530 nm, and the content was calculated using a calibration curve.

[0140] For standard products, galacturonic acid was dissolved in distilled water to prepare concentrations of 1000, 500, 250, 125 mg/L. 20 μl of the standard solution, 80 μl of distilled water, 50 μl of 0.1% carbazole ethanol reagent (prepared with 0.125 g of carbazole/100 ml of anhydrous ethanol), and 600 μl of sulfuric acid were added to a 2 ml tube and stirred. Then, 200 μl each was placed in a 96-well plate. The absorbance was measured at 530 nm to make a calibration curve, and the concentration was measured. The acidic polysaccharides content is shown below (unit: mg/g).

TABLE-US-00002 TABLE 2 Acidic polysaccharides Red ginseng 0.37 Black ginseng 2.63

[0141] As shown in Table 2, the black ginseng concentrate contained a large amount of acidic polysaccharides compared to the red ginseng concentrate.

Experimental Example 5. Analysis of Polyphenol Components in Black Ginseng Concentrate

[0142] The polyphenols content present in the red ginseng concentrate and the black ginseng concentrate obtained in Preparation Example 3 was measured using a Microplate Reader (Powerwave XS, BioTek, USA). 2 g of sodium carbonate (Sigma 223484, CAS No. 497-19-8) is taken in a 100 mL constant volume flask. Distilled water was added to adjust the total volume to 100 mL to prepare a 2% sodium carbonate reagent. Folin-Ciocalteu's phenol reagent (Sigma F9252-1L) and distilled water were mixed at a ratio of 1:1 to prepare 50% Folin-Ciocalteu's phenol reagent, which was wrapped with aluminum foil to prevent light from being transmitted.

[0143] As for the test solutions, the red ginseng concentrate and the black ginseng concentrate obtained in Preparation Example 3 each were diluted with distilled water at a ratio of 1:1. 0.1 mL of each solution was taken and mixed with 0.1 mL of 50% Folin-Ciocalteu's phenol reagent and 2 mL of 2% sodium carbonate. Then, the mixture was allowed to stand in a dark place for 30 minutes, and the absorbance was measured at 750 nm. The standard solution was prepared by taking 0.4 g of gallic acid (Sigma G7384, CAS No. 149-91-7) in a 100 mL constant volume flask, adding distilled water to adjust the total volume to 100 mL, and diluting the solution to concentrations of 31.25 ppm, 62.5 ppm, 125 ppm, 250 ppm, and 500 ppm. The standard solution was mixed with the reagents in the same manner as the test solutions. Then, the mixture was allowed to stand in a dark place for 30 minutes, and the absorbance was measured at 750 nm.

[0144] After absorbance measurement, a calibration curve was prepared with the absorbance of the standard solution as the abscissa axis and the concentration of the standard solution as the ordinate axis. The total polyphenol content of the red ginseng concentrate and the black ginseng concentrate obtained in Preparation Example 3 was calculated using Equation 1 below.

Total polyphenol content (mg/mL)=(A×B×C)/D  [Equation 1]

[0145] A: Total amount of test solution (mL), B: Dilution factor, C: Total polyphenol concentration in the test solution (mg/mL), D: Collected sample amount (mL)

[0146] The polyphenols content is shown below (unit: mg/g).

TABLE-US-00003 TABLE 3 Polyphenols Red ginseng 10.4 Black ginseng 20.8

[0147] As shown in Table 3, the black ginseng concentrate contained a large amount of polyphenols compared to the red ginseng concentrate.

Summary of Experimental Examples 3 to 5

[0148] The black ginseng concentrate prepared in Preparation Example 3 had ginsenosides Rk1 and Rg5, acidic polysaccharides, and polyphenols content higher than that of the red ginseng concentrate prepared in Preparation Example 3. Accordingly, the black ginseng concentrate of Preparation Example 3 was named as a standardized black ginseng concentrate. The above standardized black ginseng concentrate was used for measuring antiviral activity.

Experimental Example 6. Measurement of Antiviral Activity of Black Ginseng Concentrate

[0149] To measure the effect of inhibiting a new influenza virus of the standardized black ginseng concentrate prepared in Preparation Example above, an experiment was performed as follows. For each experimental group, six 6-week-old BALB/c mice (female) purchased from Samtaco Co., Ltd. were used as a group in the experiment.

[0150] The red ginseng concentrate or the standardized black ginseng concentrate were administered to mice every day at a concentration of 10 mg/Kg/day for 14 days. In addition to mice administered with the above red ginseng concentrate or the standardized black ginseng concentrate, to mice to be used as negative or positive controls, 30 μl of the new influenza virus (A/California/04/2009(H1N1)) was nasally inoculated, respectively. The red ginseng concentrate or the standardized black ginseng concentrate were administered to mice administered with the red ginseng concentrate or the standardized black ginseng concentrate for a further week after the influenza virus inoculation. The group not inoculated with the influenza virus was regarded as the normal group. Among the mice infected with the influenza virus, the sample-untreated group was set as a negative control group, and the group treated with the H1N1 influenza virus drug Tamiflu among the mice infected with the influenza virus was set as a positive control group. All groups were observed for the survival/fatality of mice for 14 days after the infection. To identify the mechanism of antiviral immunity effect of black ginseng, whether lung tissue was damaged five days after the infection was checked, and immune indicators (GM-CSF: granulocyte-macrophage colony-stimulating factor, IFN-γ: interferon-gamma, and IL-10: interleukin-10) were measured 1, 3, 5, and 7 days after the infection.

[0151] 6-1. Assessment of Fatality after Virus Infection

[0152] The fatality rate after the virus infection is shown in FIG. 3 and Table 4.

TABLE-US-00004 TABLE 4 Dosage Fatality Samples (mg/kg/day) (%) Negative control — 100 Positive control (Tamiflu) 2 0 Red Ginseng Concentrate 10 50 Black Ginseng Concentrate 10 0

[0153] As shown in FIG. 3 and Table 4, the negative control group showed 100% fatality after the virus infection. Meanwhile, the fatality rate of the positive control group treated with the antiviral drug Tamiflu was 0%. In the red ginseng concentrate-administered group, a total of 3 mice out of 6 mice used in the experiment died from the virus infection, resulting in a 50% fatality rate. However, a total of six mice in the standardized black ginseng concentrate-administered group all survived. Thus, the fatality rate was 0%, the same as that of the positive control group. Therefore, the protective effect of the standardized black ginseng concentrate against the H1N1 influenza virus was significantly superior to that of the red ginseng concentrate.

[0154] 6-2. Assessment of Lung Tissue Damage Degree after Virus Infection

[0155] The cause of death of mice infected with the H1N1 influenza virus means loss of respiratory function caused by tissue damage due to the accumulation of immune cells in the lung tissue 5 days after the infection, as in the lung tissue of the negative control group or the red ginseng concentrate-administered group shown in FIG. 4. The standardized black ginseng concentrate-administered group showed relatively low lung tissue damage degree compared to the red ginseng concentrate-administered group. The standardized black ginseng concentrate was shown to be superior to the red ginseng concentrate in inhibiting or ameliorating lung tissue damage caused by the H1N1 virus infection.

[0156] 6-3. Assessment of Immune Activator Production Degree after Virus Infection

[0157] The effective treatment step for the H1N1 influenza virus is that immune cells proliferate at the initial stage of infection and produce a large number of immune activation factors to effectively remove the virus, and at the late stage of infection, immune cells produce immune inhibition factors to restore the host to a normal state.

[0158] As a result of this experiment, there was no change in the production of immune activation factors in the normal group, as indicated by the dotted line in the graph. Meanwhile, in the standardized black ginseng concentrate-administered group, the immune cell proliferation factor (GM-CSF) was highly expressed on day 1 of infection, the immune activation factor (IFN-γ) was highly expressed on day 3 of infection, and the immune inhibition factor (IL-10) was highly expressed on day 7 of infection. Therefore, the standardized black ginseng concentrate was found to exert an antiviral effect by increasing the proliferation and activity of immune cells at the initial stage of infection to kill the virus and then normalizing the raised immune system at the late stage of infection (see FIGS. 5a to 5c). In addition, most of these immune activation factors showed higher activity compared to the red ginseng concentrate-administered group, indicating that the standardized black ginseng concentrate had higher therapeutic and preventive effects on the H1N1 virus infection than the red ginseng concentrate.

[0159] This is considered to be because the standardized black ginseng concentrate has a higher content of ginsenosides Rk1, Rg5, acidic polysaccharides, and polyphenols than the red ginseng concentrate.

Experimental Example 7. Measurement of the Effectiveness of Black Ginseng Concentrate on Respiratory Health

[0160] To measure the effectiveness of the standardized black ginseng concentrate prepared in Preparation Example above on respiratory health, an experiment was performed as follows. For each experimental group, six 6- to 8-week-old C57BL/6 mice (male) were used as a group in the experiment.

[0161] The red ginseng concentrate or the standardized black ginseng concentrate were administered to mice every day at a concentration of 10 or 250 mg/Kg/day for 14 days. 50 μl of ultrafine dust (SRM2975 10 mg/ml) was administered intratracheally for three days every day to mice to be used as the negative control group as well as mice administered with the red ginseng concentrate or the standardized black ginseng concentrate. The red ginseng concentrate or the standardized black ginseng concentrate were administered to mice administered with the red ginseng concentrate or the standardized black ginseng concentrate for a further week from the day of ultrafine dust administration. The group to which ultrafine dust was not administered was regarded as the normal group. All groups were euthanized 4 to 6 days after the initial injection of ultrafine dust. Then, the weight of the lungs and the number of immune (inflammatory) cells were measured to determine the anti-inflammatory effect of the black ginseng. A histopathological examination of the lungs was performed to determine the effect of reducing ultrafine dust accumulation in the respiratory lung of the black ginseng. In addition, to investigate the mechanism of respiratory health function against ultrafine dust of the black ginseng, immune indicator factors (GM-CSF: granulocyte-macrophage colony-stimulating factor, TNF: tumor necrosis factor, IL-1beta: interleukin-1beta, IL-6, IL-2, & IL-10) were measured.

Experimental Example 8. Measurement of Anti-Aging Function of Black Ginseng Concentrate

[0162] An experiment was performed to measure the effectiveness of the standardized black ginseng concentrate prepared in Preparation Example above on anti-aging function, as follows. For each experimental group, three 18-month-old C57BL/6 mice (male) were used as a group in the experiment.

[0163] The red ginseng concentrate or the standardized black ginseng concentrate were administered to mice every day at a concentration of 300 mg/Kg for 14 days. All groups were injected every day for 28 days and then euthanized. To confirm the anti-aging effect of black ginseng, the expression of genes and proteins, such as aging-related indicators p15INK4b, p16INK4a, p21, p27, p38, p53, CDK1, CDK2, mechanistic target of rapamycin (mTOR), sirtuin 1 (SIRT1), and β-galactosidase (β-gal) staining were measured from liver tissues and hepatocytes isolated from liver tissues.

Preparation Example 4. Preparation of Foods

[0164] 4-1. Preparation of Wheat Flour Foods

[0165] 0.5 to 5.0 parts by weight of the black ginseng extract of the present invention was added to wheat flour, and this mixture was used to prepare bread, cakes, cookies, crackers, and noodles.

[0166] 4-2. Preparation of Soups and Gravies

[0167] 0.2 to 5.0 parts by weight of the black ginseng extract of the present invention was added to soups and gravies to prepare meat-processed products for health improvement, noodle-style soups, and gravies.

[0168] 4-3. Preparation of Ground Beef

[0169] 10 parts by weight of the black ginseng extract of the present invention was added to ground beef to prepare ground beef for health improvement.

[0170] 4-4. Preparation of Dairy Products

[0171] 5 to 10 parts by weight of the black ginseng extract of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

[0172] 4-5. Preparation of Dry Cereal

[0173] Brown rice, barley, glutinous rice, and adlay were pregelatinized by a conventional method, dried, roasted, and prepared into powders by a pulverizer to have a particle size of 60 mesh.

[0174] Black beans, black sesame, and perilla seeds were also steamed and dried by a conventional method, roasted, and prepared into powders by a pulverizer to have a particle size of 60 mesh.

[0175] The black ginseng extract of the present invention was concentrated under reduced pressure, sprayed, and dried by a hot air dryer. The resulting dry product was prepared into powders by a pulverizer to have a particle size of 60 mesh.

[0176] The thus-prepared grains, seeds and nuts, and the black ginseng extract of the present invention were mixed in the following ratio to prepare dry cereal:

[0177] grains (brown rice (30 parts by weight), adlay (15 parts by weight), and barley (20 parts by weight)), seeds and nuts (perilla seeds (7 parts by weight), black beans (8 parts by weight), black sesame (7 parts by weight), the black ginseng extract of the present invention (3 parts by weight), Ganoderma lucidum (0.5 part by weight), and Rehmannia glutinosa (0.5 part by weight).

[0178] 4-6. Preparation of Health Drinks

[0179] 5 g of the black ginseng extract of the present invention was homogeneously mixed with minor ingredients, such as high fructose corn syrup (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), and water (75%). The mixture was sterilized instantly and packaged in small containers, such as glass bottles, pet bottles, etc., to prepare health drinks.

[0180] 4-7. Preparation of Vegetable Juice

[0181] 5 g of the black ginseng extract of the present invention was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice.

[0182] 4-8. Preparation of Fruit Juice

[0183] 1 g of the black ginseng extract of the present invention was added to 1,000 ml of apple or grape juice to prepare fruit juice.

Preparation Example 5. Preparation of Pharmaceutical Compositions

[0184] 5-1. Preparation of Powders

[0185] The black ginseng extract of the present invention 2 g

[0186] Lactose 1 g

[0187] The above ingredients were mixed and filled in an airtight pouch to prepare powders.

[0188] 5-2. Preparation of Tablets

[0189] The black ginseng extract of the present invention 100 mg

[0190] Corn starch 100 mg

[0191] Lactose 100 mg

[0192] Magnesium stearate 2 mg

[0193] The above noted ingredients were mixed and tabletted according to a conventional tablet preparation method to provide tablets.

[0194] 5-3. Preparation of Capsules

[0195] The black ginseng extract of the present invention 100 mg

[0196] Corn starch 100 mg

[0197] Lactose 100 mg

[0198] Magnesium stearate 2 mg

[0199] The above ingredients were mixed and filled in a gelatin capsule according to a conventional method for preparing capsules to prepare capsules.

[0200] 5-4. Preparation of pills

[0201] The black ginseng extract of the present invention 1 g

[0202] Lactose 1.5 g

[0203] Glyserine 1 g

[0204] Xylitol 0.5 g

[0205] The above ingredients were mixed and prepared into a pill according to a conventional method in such a manner that one pill has a weight of 4 g.

[0206] 5-5. Preparation of Granules

[0207] The black ginseng extract of the present invention 150 mg

[0208] Soybean extract 50 mg

[0209] Glucose 200 mg

[0210] Starch 600 mg

[0211] The above ingredients were mixed and 100 mg of 30% ethanol was added thereto, followed by drying at 60° C. After formation of granules, the granules were filled into packaging.