KR20140081013A - (Composition comprising the extract of crude drug complex for promoting bone growth) - Google Patents

Abstract

The present invention relates to a composition comprising a crude drug complex extract for promoting the growth of bone as an active ingredient and, more specifically, to a composition for promoting growth of bone, which contains the crude drug complex extract consisting of red ginseng, atractylodes koidzmi, eleutherococcus senticosus, cornus cervi parvum, eucommia ulmoides oilver, astragali radix, hoehen, cortex fraxini, angelicae gigantis radix, and radix glycyrrhizae. The crude drug complex extract effectively promotes the growth of bone, thereby being utilized for promoting the growth of children.

Description

The present invention relates to a composition for promoting bone growth comprising an extract of crude drug complex as an active ingredient,

The present invention relates to a composition for promoting bone growth comprising an extract of crude drug complex as an active ingredient, and more particularly, to a composition for promoting bone growth comprising an extract of a crude drug complex made of Rhizoma, Agaricus, To a bone growth promoting composition capable of promoting bone growth of children and adolescents in growing period.

The growth of children and adolescents due to economic growth and income level is of great interest. The increase in the level of nutrition and the improvement of the living environment and lifestyle, the average height and weight of puberty It is gradually improving. As the advantages of such height increase, various methods for promoting growth are emerging.

Growth involves not only an increase in body length and weight, but also an increase in the anatomical and morphological size of the organs of the body. However, in general, when measuring the growth of children and adolescents, the size of the kidney is mainly used as an index. The increase of the kidney is the skeletal metabolism including the synthesis of the cartilage tissue by the nutrition and the growth hormone, the growth of the skeletal length and the skeletal tissue proliferation. Especially, the growth hormone which is a specific substance that promotes the growth of the body, Length is one of the important substances that affect growth and growth of muscles and organs of all tissues.

Factors affecting growth include normal childhood growth and inherited factors. In particular, the genetic factor is 23%, which is determined by the remaining 77% (31%), exercise (20%), environment (16%) and others (10%). Therefore, in order to promote growth, nutrition of balance, proper exercise, and environmental improvement are required, but nutrition of balance is pointed as the most important factor. In particular, irregular eating habits and uncomplicated eating and frequent fast food and fast food intake are leading to overweight and childbearing children, and underweight children due to poor eating, irregular eating times and poor nutrient intake also increase due to social problems (Lee, KH et al., Korean J Community Nutrition, 8, 3-14, 2001).

In order to balance these effects, growth - promoting health functional foods are emerging, but the effect has not reached a satisfactory level due to the combination of various raw materials which are merely useful for growth.

Therefore, the inventors of the present invention have found that it is possible to improve the osteoblast differentiation ability through the herbal composition extract which does not affect the body metabolism of the recipient while promoting the growth, escaping from the simple combination of the ingredients having growth promoting effect, Intestine length, growth hormone, and IGF-I, and thus completed the present invention.

Accordingly, an object of the present invention is to provide a composition for promoting bone growth comprising an extract of crude drug complex as an active ingredient.

It is also an object of the present invention to provide a health functional food which can help promote growth.

In order to attain the above object, the food showing the growth promoting effect of the present invention may be one or more selected from the group consisting of 1 to 20 drops of water, 10 to 30 drops of visa apricot, 5 to 20 drops of antler, 5 to 20 drops of antler, 20 to 60 drops of antler, 5 to 20, Angelica keiskei (capsule) of 5 to 20, and Licorice (licorice) 1 to 10 weight ratio (w / w) as an active ingredient.

In addition, the food showing the growth promoting effect according to another comparative example of the present invention is characterized in that it contains an alcohol extract or a hot-water extract containing 100 parts by weight (w / w) of red ginseng as an active ingredient.

As described above, since the composition of the present invention has an effect of promoting bone growth, it is very useful as a functional food and a health food of children / adolescents during growing period.

1 is a graph showing the cytotoxicity of osteoblasts according to the concentration of the composition.
2 is a graph showing the activity of basic alkaline phosphatase according to the concentration of the composition.
A: Red ginseng extract diet;
B: Ingredients extracted with herbal medicine composite extract;
C: Red ginseng extract: crude drug complex extract = 1: 1 dietary group;
D: Red ginseng extract: crude drug complex extract = 2: 1 dietary group; And
E: Red ginseng extract: crude drug composite extract = 3: 1 diet group.
*: P < 0.05
**: P < 0.01
3 is a graph showing the secretion promoting activity of growth hormone (GH) on the composition.
Figure 4 is a graph showing the secretion promoting activity of IGF-I on the composition.
**: P < 0.01

The active ingredients of the composition of the present invention are hot-water extracts of herbal medicines composed of Paeonia, Rhodophyta, Deer, Dioptera, Hwanggi, Baekbokryeong, Dhamma, Angelica gigantosa and Licorice. Each herbal medicine component constituting the herbal medicine extract has the following effects I have.

Atractylodes macrocephala Koidzumi has been reported to increase the weight gain and endurance of mice, to increase phagocytosis, and to promote cellular immune function. It also has a pharmacological action such as controlling the intestinal (intestinal) inhibitory action and excitatory action, anti-ulcer and liver function protecting action, immune function hyperactivation action, vasodilating action, diuretic action and hypoglycemic action,

 It is believed that Eleutherococcus senticosus (Rupr. & Maxim.) Maxim., Acanthopanax senticosus (Rupr. &Amp; Maxim.) Harms.) Has a pharmacological effect such as antiinflammatory, analgesic, antistress, immunosuppressive, I have.

 Cornus cervi parvum has been used for the purpose of gangjeong and tannage and has pharmacological actions such as growth promoting action, hematopoietic action, protein synthesis promoting action, cholesterol lowering action and immunity increasing action.

 Eucommia ulmoides Oliver is an antioxidant, anti-fatigue, glucosidase inhibitor, vasodilator, anti-diabetic, analgesic and neuralgia, protein synthesis and lipid metabolism promoting action, Absorption and prevention of bone mineral loss, and a significant effect on osteogenesis.

 Astragali Radix is a medicine for the treatment of physical weakness, sweating, swelling, stomachache, abdominal pain, kidney damage, ear ache, 虛 喘, (痛 痛), food tasteless, lack of appetite, hemorrhoids, purpose (赤)), school illness (疾 虫), abortion, uterine bleeding, wife manslaughter, menstrual irregularities,祛痰 软), headache, deep mulching (肺 pulmonary heartburn), dysentery and pediatric diseases have pharmacological effects.

 Hoelen, Poria cocos (FR.) WOLF) has pharmacological actions such as diuretic action, hypoglycemic action, increase in the contractility of the heart, and immunity enhancement.

 Cortex Fraxini contains volatile essential oils, glycosides Hesperidin (C28H34O15), citric acid, etc. It releases loose aggregates and strengthens the function of the spleen, so that only the abdominal cavity, trimming, vomiting, nausea, indigestion, Symptoms, thinning of the stool is treated. In addition, the essential oil component has a pharmacological action such as digestive stimulation, digestive stimulation, germane, anti-ulcer, antidiarrheal secretion, coronary artery, hypertension, hyperalgesia, promotion of bile secretion, inhibition of uterine smooth muscle and antibacterial action.

 Angelicae Gigantis Radix (Root of Angelica gigas Nakai (Umbelliferae)) is an antioxidant that acts on the blood and hematopoietic system, on the uterus, on the cardiovascular system, on the immune system, on the analgesic action, Antimicrobial action, liver function-protecting action, anti-cancer action and hematopoietic action.

 Radix Glycyrrhizae contains glycyrrhizin (C42H66O18), other liquiritin (C21H22O9), and asparagine. It harmonizes the toxicity of all drugs and makes them appear more effective. It regulates the heat and scent of the books, Good communication and strong muscles and bones. Jinhae genitour, muscle relaxation, diuretic, anti-inflammatory and suppresses peptic ulcers. It also has pharmacological actions such as detoxification, hepatitis, urticaria, dermatitis, and eczema.

The composition for promoting bone growth of the present invention may be used in combination with the above-mentioned effective amount of 1 to 20, 10 to 30, 30 to 20, 5 to 20, 2 to 5, 20 to 60, 5 to 20, 5 to 20, (W / w) of licorice and 5 to 20 of licorice, and 1 to 10 weight ratio (w / w) of licorice, and then extracting with hot water as an active ingredient.

The method for obtaining the crude drug composite material extract of the present invention is as follows.

Distilled water, especially tertiary distilled water, containing no impurities is added to the herbal medicine mixture, and the extract is extracted from 80 to 100 hours for 2 hours to 4 hours.

If the temperature of the hot water extraction is lower than 80 or the time is shorter than 2 hours, the active ingredient is not eluted well. If the temperature is higher than 100 or the time exceeds 4 hours, the active ingredient may be decomposed. It is preferable to extract hot water in the range of time. As the extraction method, hot water extraction, cold extraction, reflux cooling extraction or ultrasonic extraction are preferable, and more preferably, the extract of the present invention can be obtained by heating under reflux and concentration under reduced pressure. The herbal composition extract of Bacillus thuringiensis, Bacillus thuringiensis, Deer, Deer, Hwanggi, Baekbokryeong, Dermis, Angelica and Licorice was named I09.

The herbal medicine composite extract of the present invention can be used for various purposes by extracting hot water, extracting the extract by filtration, or lyophilizing the concentrated extract.

The composition of the present invention can be prepared in the form of a food added to various foods and exhibiting a bone growth promoting effect. For example, there are various foods, beverages, tea, candies, gums, vitamin complexes, health functional foods and the like, and they are formulated in the form of powders, capsules, powders, tablets, .

In general, the composition of the present invention preferably contains 0.01 to 99.9%, more preferably 0.1 to 50%, most preferably 1 to 25%, and most preferably 100 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 mg / ml.

The food of the present invention promotes the growth of the skeleton effectively without affecting the metabolism in the body, and thus can be widely utilized for promoting bone growth of infants and adolescents.

Hereinafter, the present invention will be described in more detail by way of examples.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

< Example  1 to 2> crude drug composite extract ( I09 ) And red ginseng extract

500 g of white liquorice, thorns acne, antler, duchung, hwanggi, Baekbokryeong, duck, ginseng and licorice

The weight was measured so as to have the composition and content ratio as shown in Table 1. Distilled water corresponding to 1 part of the weight of the sample was added, heated and refluxed for 4 hours, and filtered under reduced pressure to separate the extract and the residue. The extract (hereinafter referred to as I09) obtained by concentration under reduced pressure using a rotary evaporator under reduced pressure was used as a sample in the following experiment.

The red ginseng extract was extracted at 55-65 hours for 10 hours by adding 8 times (w / w) of 70% ethanol (30% water) to 100% hot water extract. The extracts were each concentrated at 40-45 to be 65 Brix.

Furtherance ingredient
Example 1
Example 2
Weight (g)
content(%)
Weight (g)
content(%)
Red ginseng
50
100
0
0
Boiled
0
0
5
10
Visible
0
0
8
16
velvet
0
0
4
8
Mushrooms
0
0
4
8
Hwanggi
0
0
15
30
Baekbokryung
0
0
4
8
dermis
0
0
4
8
Angelica
0
0
4
8
licorice
0
0
2
4

< Example  3 to 5> crude drug complex extract ( I09 ) And red ginseng extract

The compositions of the composition ratios shown in Table 2 below were prepared by mixing Examples 1 and 2 as follows. In order to compare the growth effect of the present invention, saline was used as a control group.

Furtherance Red ginseng extract
Herbal medicine composite (I09)
Example 3
Weight (g)
30
30
Weight portion
One
One
Example 4
Weight (g)
40
20
Weight portion
2
One
Example 5
Weight (g)
45
15
Weight portion
3
One

< Experimental Example  1> Measurement of osteoblast activity

<1-1> Culture of osteoblast

Human osteoblast-like cell line MG-63 (Seoul National University College of Medicine, Seoul, Korea) was purchased and used. DMEM containing 10% fetal bovine serum (Dulbeccos modified Eagles medium, and cultured in a 37% incubator containing 5% CO ₂ . MG-63 was added to a 24-well plate at 210 ³ cells / well and cultured for 24 hours.

<1-2> Cytotoxicity measurement

After completion of the culture, CCK-8 (Dojindo laboratory, Japan) assay was used to measure the toxicity of the composition. That is, MG-63 cells were seeded at a density of 510 ³ cells / well in a 96-well plate. After incubation in an incubator at 37 and 5% CO ₂ , 10 different concentrations of

The control and the compositions of Examples 1 to 2 and Examples 3 to 5 were treated at a concentration of 62.5, 125, 250, 500 / and cultured for 48 hours. Cell viability was determined by using CCK-8 kit for 48 h culture time-normal cells.

The experimental results are shown in Fig. As can be seen from Fig. 1, it was confirmed that there is no or very little cytotoxicity at the above concentrations.

< Experimental Example  2> Measurement of basic phosphatase activity

Alkaline phosphatase (ALP) is an enzyme that binds to the cell membrane of osteoblast cells and is used as a labeling enzyme for osteoblasts. Specifically, the basic phosphatase activity test is performed using p-nitrophenyl phosphate : a method of calculating the concentration of basic phosphatase by measuring the amount of p-nitrophenol, which is a hydrolysis product, by using a basic phosphatase as a catalyst for the hydrolysis reaction of pNPP to be.

MG-63 osteoclasts were seeded at a density of 110 ⁵ cells / well in a 6-well plate and cultured in a 37 ° C CO ₂ incubator for 24 hours. The medium was removed and 10% fetal bovine serum (FBS) % New antibiotic (penicillin-streptomycin) was added to the culture medium. After culturing for 2 days, the culture was removed and the cells were treated with 0.1% Triton X-100 (Triton X-100). The cell suspension was centrifuged at 3,000 rpm for 3 to 10 minutes, and the supernatant was taken for ALP activity measurement. ALP was measured by adding 1M Trizma base buffer (pH 10) 150, 5 mM MgCl ₂ 30 and 5 mM pNPP 30 to cell lysate 90 in a 96-well plate and reacting at 37 for 1 hour to measure the amount of pNPP in a microplate reader ), Colorimetric determination was made at 405 nm, and the enzyme activity per unit protein content was measured by dividing by the amount of protein. Protein content was measured by Bradford method using bovine serum albumin (BSA), and the enzyme activity was expressed as a percentage of the control.

The experimental results are shown in Fig. As can be seen from FIG. 2, the activity values of ALP vary depending on the composition ratio of red ginseng extract and crude drug composite extract (see Examples 3 to 5 and Table 2) at a concentration of 250 / , The ratio of ALP activity was 123.3 when the composition ratio of red ginseng extract and crude drug complex extract was 3: 1, 143.9 when 2: 1, 123.3 when 1: 1, and 111.6 when 0: , And when it is 1: 0, it is 103.9.

From this, it can be deduced that the ALP activity is significantly increased when the composition ratio of red ginseng extract and crude drug composite extract is 1: 1 to 2: 1. In other words, when the red ginseng extract and crude drug complex extract were mixed at a composition ratio of 1: 1 to 2: 1 compared to the case where the red ginseng extract and the crude drug composite extract were separately added, .

These results indicate that the composition of the extract of red ginseng extract and crude drug complex promotes osteoblast differentiation because the basic phosphatase is a biomarker that is expressed in differentiation of osteoblasts.

< Experimental Example  3> Growth promoting efficacy analysis

<3-1> Preparation of test animal models

Three-week-old ICR mice (male) were purchased and adapted for 5 days at a constant temperature (25) and humidity (505%).

<3-2> Test group selection and sample treatment

In order to examine the growth promoting effect by the addition of the composition, the composition of each of the untreated control group and red ginseng extract and herbal medicine composite extract (Examples 1 to 2 and Example 4) was changed to 0.8% (w / w) Mixed diets were fed for 5 weeks. Eight mice were used in each test group.

<3-3> Analysis of weight gain and feed intake

As shown in the following Table 3, the weight gain and feed intake of the control group, the red ginseng extract and the herbal composition extract group were significantly improved compared to the control group, and the intake of the red ginseng extract: crude drug extract = 2: 1 Respectively.

Test group
Initial weight (g)
Final weight (g)
Weight gain (g / week)
Feed intake (g / day)
Control group
15.62.8
37.20.5
4.10.1
4.50.9
Red ginseng extract
16.11.3
40.74.4
4.80.8
4.95.0
Herbal medicine composite extract
15.51.3 ^*
41.62.3 ^**
5.10.5 ^*
4.71.7
Red ginseng extract: crude drug complex extract = 2: 1
15.41.2 ^*
37.03.9
4.40.6
5.85.5 ^*

^* : P < 0.05

^** : P < 0.01

<3-4> Analysis of growth effect of iliac bone

Each test group was sacrificed for 5 weeks after feeding for 5 weeks. Eight of each group were sacrificed for 5 weeks. The right iliac bone was removed and the length of the iliac bone was measured using a caliper. The results are shown in Table 4 below.

As shown in Table 4, the red ginseng extract and crude drug composite extract (Examples 1 to 2 and Table 1) and the red ginseng extract and crude drug composite extract mixture (Examples 3 to 5 and Table 2) treatment groups were significantly higher than those of the control group .

Growth rate (%) = 100 + (length growth control group length growth) / control length growth 100

division
Length of iliac bone (mm)
Growth rate (%)
Control group
16.310.05
100
Red ginseng extract
16.920.13 ^*
103.8
Herbal medicine composite extract
16.910.28 ^*
103.7
Red ginseng extract: crude drug complex extract = 2: 1
16.880.03 ^**
103.5

^* : P < 0.05

^** : P < 0.01

<3-5> Analysis of GH and IGF-I in blood

Rats were anesthetized and sacrificed after 5 weeks of feeding the samples in each test group. Blood was collected and centrifuged at 3,000 rpm for 12 minutes to separate the serum. Separated serum growth hormone (GH) and IGF-I content were measured using an OCTEIA Rat / Mouse IGF-I kit (Immunoenzymometric assay: IEMA).

The experimental results are shown in FIGS. 3 and 4. FIG. As can be seen from FIG. 3, the concentration of red ginseng extract: crude drug complex extract = 2: 1 was higher than that of the control group.

As shown in FIG. 4, the IGF-I concentration in the blood was significantly increased in the experimental group as compared with the control group.

Claims (4)

A composition for accelerating bone growth, comprising as an active ingredient an extract of a herbal medicine composite comprising red ginseng extract and alfalfa, astragalus, antler, japonica, hwanggi, baekbokryeong, dermis, ginseng and licorice.
The composition for promoting bone growth according to claim 1, wherein the composition for promoting bone growth comprises 1 to 20 parts by weight of total weight, 10 to 30 parts by weight of viscose, 5 to 20 parts by weight of antler, 5 to 20 parts by weight, 20 to 60 parts by weight of Huang, 5 to 20 parts by weight, 20, Angelica gigas 5-20, Licorice 1 ~ 10%.
[3] The composition for promoting bone growth according to claim 1, wherein the composition ratio of the red ginseng extract and the crude drug composite extract is 1: 1 to 2: 1.
A health functional food for promoting bone growth comprising the composition of any one of claims 1 to 3.

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