KR101777040B1

KR101777040B1 - the GamiSibjeondaebo composite with the funtion of the treatment of scull fracture

Info

Publication number: KR101777040B1
Application number: KR1020160018420A
Authority: KR
Inventors: 김동희; 최학주; 심부용; 박지원; 이효정; 주인환
Original assignee: 대전대학교 산학협력단
Priority date: 2016-02-17
Filing date: 2016-02-17
Publication date: 2017-09-08
Also published as: KR20170096758A

Abstract

The present invention relates to a natural composition having a function of restoring fracture, and more particularly, to a natural composition having an effect for restoration of skull fracture including ginseng, alopecia, gypsum, licorice and the like.
In order to solve the above problems and needs,
The present invention provides an anticancer composition which is effective for recovering a skull fracture extracted from a mixture of ginseng, red ginseng root, ginseng root, licorice root, peony root, peony root, Angelica gigas,
The present invention also relates to a method for producing ginseng, which comprises adding 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of licorice, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, , 80-120 parts by weight of Angelica gigas Nakai, 80-120 parts by weight of Hwanggi, 80-120 parts by weight of broth, 120-180 parts by weight of natural copper, 120-180 parts by weight of MB, 120-180 parts by weight of skeleton bone, Which is effective for recovery, is provided.
In addition, the present invention provides a pharmaceutical composition comprising a potent anti-hypertensive agent which is effective for restoring the skull fracture.

Description

[0001] The present invention relates to a composition for the treatment of fractures of the skull,

The present invention relates to a natural composition having a function of restoring fracture, and more particularly, to a natural composition having an effect for restoration of skull fracture including ginseng, alopecia, gypsum, licorice and the like.

Fractures are those that can cause pain, swelling, soft tissue damage, and bruises due to internal bleeding, which means that the bones are cracked or cracked. Anyone, regardless of age, race or economic background, is vulnerable to fractures. Fractures are most often caused by physical trauma, such as a car accident, physical abuse, or severe falls. However, the lower the bone mineral content, the greater the likelihood that a person will fracture.

For example, damage associated with osteoporosis is a significant medical challenge for elderly people with increased bone fragility, especially because the risk of falls due to accidents is greater. Fractured hips, wrists, and spine are among the most common injuries associated with osteoporosis. In particular, hip fracture is extremely uncomfortable and costly, and in women, it also correlates with high mortality and morbidity.

In most cases, fractures are treated with fixation with a pad, plaster cast, or fixture, while restricting activity for an extended period of time. The effect of fracture treatment on patients and their families is significant. The direct costs of fracture treatment usually include hospital costs and physiotherapy costs. In the United States in 1995, medical expenses related to the treatment of forearm fractures amount to $ 385 million (Ray et al., J. Bone Miner Res., 12 (1): 24-25 (1997)). Indirect costs are more difficult to estimate.

Fractures usually limit productivity and, in some cases, reduce opportunities for money and the ability to care for family members. Fractures also affect the quality of life and pride of the patient. For example, survivors of hip fractures have reported a 52% reduction in quality of life after 12 months of fracture (Tosteson et al., Osteoporos Int., 12 (12): 1042-49 (2001) .

The prior art and prior art described above are still toxic to the liver and kidney and are insufficient for the treatment of skull fracture recovery.

The present invention seeks to provide a composition that is remarkably effective in the recovery of skull fracture without toxicity to the liver and kidney.

The present invention also provides a composition having significant efficacy in the recovery of skull fracture which is effective for antioxidant and anti-inflammation.

In order to solve the above problems and needs,

The present invention provides an anticancer composition which is effective for recovering a skull fracture extracted from a mixture of ginseng, red ginseng root, ginseng root, licorice root, peony root, peony root, Angelica gigas,

The present invention also relates to a method for producing ginseng, which comprises adding 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of licorice, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, , 80-120 parts by weight of Angelica gigas Nakai, 80-120 parts by weight of Hwanggi, 80-120 parts by weight of broth, 120-180 parts by weight of natural copper, 120-180 parts by weight of MB, 120-180 parts by weight of skeleton bone, Which is effective for recovery, is provided.

In addition, the present invention provides a pharmaceutical composition comprising a potent anti-hypertensive agent which is effective for restoring the skull fracture.

The present invention relates to a composition for preventing and treating skull fracture, which is effective for restoration of skull fracture according to the present invention, although it has no toxicity to liver and kidney.

In addition, the anticancer composition having the effect of restoring the skull fracture according to the present invention has a remarkable effect on antioxidant and anti-inflammation.

FIG. 1 is a photograph of the X-ray photograph taken at intervals of 4 weeks from the 0th to 8th weeks of the cranial fracture, which proves the effect of the composition according to the present invention on the union of the skull fracture.
FIG. 2 is a photograph of a micro-CT image obtained after the experiment to confirm the effect of administration of the composition according to the present invention on the union of the skull fracture and then converting the image into 3D.
FIG. 3 is a graph showing bone mineral density (BMD) measured by 3D micro-CT analysis of skulls according to administration of the composition according to the present invention and calculating BMD (bone mineral density).
FIG. 4 is a graph showing bone area obtained by calculating the ratio of bone volume to BV by analyzing the skull with 3D micro-CT by administration of the composition according to the present invention.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition which is effective for restoration of skull fracture extracted from a mixture of ginseng, white liquor, gyeongryeong, licorice, sorghum, peony, celeste, angelica, hwanggi, broiler, natural copper,

The present invention is referred to as GamiSibjeondaebo-Tang (hereinafter referred to as GST ) composition.

Preferably, the present invention is a ginseng composition comprising 80 to 120 parts by weight of potash, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of licorice, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, , 80-120 parts by weight of Angelica gigas, 80-120 parts by weight of broilers, 120-180 parts by weight of natural broth, 120-180 parts by weight of alum husks, 120-180 parts by weight of bone marrow, Thereby providing a composition that is effective in restoring skull fracture.

In the present invention, the above raw materials are mixed with the above-mentioned weight units, and 500-2000 parts by weight of purified water is mixed with 100 parts by weight of the whole raw materials to obtain an extract.

In addition, the present invention relates to a process for producing a concentrate by concentrating 100 parts by weight of a raw material mixed with the above materials in an amount of 1000 to 2000 ml of 70 to 90% alcohol (alcohol) and refluxing for 2 to 5 hours, filtering the resulting mixture by rotary evaporation using a rotary vacuum evaporator can do.

The ginseng, white ginseng, ginseng, licorice, sorghum, peony, celeste, angelica, and hwanggi according to the present invention are used as common medicinal materials.

The above broiler system of the present invention is a stem skin of Cinnamomum cassia Presl (camphora and Lauraceae), or it is removed as it is. Broiler chickens may have different origins and different qualities depending on the area. In particular, Vietnam has divided broilers into five classes. In addition, since broiler chickens have different years of harvesting and processing methods, the product standard is relatively comparable in terms of varieties (邊 계, 官, 판, 계, 계, and 桂) many.

The above natural copper of the present invention is a sulfurized mineral pyrite. It is calculated as hexagonal crystals, and some of them are produced as ellipsoids with high roundness due to erosion by water. This implies that the origin of the medicinal material is two mixtures: weathered residues at the site and transported sediments. The hexagonal and ellipsoidal surface of the particles is changed to iron oxide by weathering, but yellowish white pyrite is observed in the inside of the crushed metal. Outer surface is bright and pale yellow with metallic luster but yellowish brown to brown with no metallic luster. There is a bar pattern, and the mark of the bar is black to red to reddish brown. It is heavy and the vagina is hard and hard, but it is a little weak and easily broken. The cut surface is yellowish white and metallic. Or the cut surface is brown, you can see a bright white star point. Hardness 6.0 to 6.5, specific gravity: 4.9 to 5.2. There is a slight peculiar smell and sour taste.

The other names of the above-mentioned mushrooms of the present invention are night skins, mixed skins, and mixed skins. Albizzia julibrissin, a silkworm, a leguminous plant, is the dried shell of Durazz. Silkworm trees are planted in various places. Between summer and autumn, the skin is peeled and dried in the sun. The taste is sweet and the quality is plain. It acts on the heart (heart), necrosis (脾经), menopause (肺经). It stabilizes the mind, promotes blood circulation, calms the swelling, stops the pain, and connects the muscles and bones. Amnesia, insomnia, pulmonary abscess, swelling, and fracture. 6 ~ 9g a day in the form of gangjejeol powder and eat. When you use it as an external medicine, put it on the base (medicine) agent with powder.

The above-mentioned bone-repairing tool of the present invention refers to the root stem of Drynaria fortunei Smith in Korea. In China, it was the same as our country, and in Japan, it was not treated as a process medicine.

The above-mentioned pharmaceutical composition means a drug which is effective for restoration of skull fracture, and it is meant that the present invention is comprised of a tamoxifen preparation.

The present invention relates to a process for preparing a mixture of 8 g of ginseng, 8 g of liquorice, 8 g of licorice, 8 g of licorice, 8 g of peony root, 8 g of peony root, 8 g of ginseng, 8 g of Angelica gigas, 8 g of Hwanggi, 8 g of broth, 12 g of natural copper, 1000 ml) was added and the mixture was refluxed for 3 hours. The filtrate was collected and concentrated under reduced pressure on a rotary vacuum evaporator.

The concentrated solution was lyophilized with a freeze dryer to obtain 4.6 g of powder. The obtained powder was stored in an ultra-low temperature freezer (-80 ° C.), and a composition (GST or GST extract) diluted in distilled water at a necessary concentration was prepared And the following experimental results were obtained.

1. Animal test method

SD rats (310-360 g) of 11-week-old male animals were supplied from Gyeonggi-do, Korea and fed with solid feed (Purina) and water until the day of the experiment. 55 ± 15%, 12 hours-12 hours (light-dark cycle) for 2 weeks. This experiment was conducted in accordance with the animal ethics code of the Daejeon National Animal Experimental Ethics Committee (Animal Use Ethics Committee Approval No. DJUARB 2015-022).

2. Effect on liver function

(1) To measure AST and ALT activity in the serum, blood was collected using the cardiac puncture method after the end of the experiment. Blood was solidified at room temperature for 30 minutes, centrifuged at 3,000 rpm for 15 minutes, and the serum was separated and analyzed by the Department of Clinical Pathology, Daejeon, Korea. AST and ALT activities were measured by a biochemical automatic analyzer using the principle of JSCC UV method.

(2) Aspartate Aminotransferase ( AST ) content

The mean AST of the liver function test was 164.0 ± 42.9 U / L in the normal group, 115.6 ± 25.5 U / L in the control group, 160.0 ± 34.0 U / L in the GST 200 ㎎ / Kg, and 143.3 ± 45.7 U / L, respectively, which showed no difference compared to the normal group.

(3) Alanine Aminotransferase (ALT) content

The ALT was measured as 50.3 ± 4.0 U / L in the normal group, 39.2 ± 13.7 U / L in the control group, 26.0 ± 8.9 U / L in the GST 200 ㎎ / ㎏ group and 400 ㎎ / ㎏ was 23.3 ± 11.5 U / L, which showed no difference compared to the normal group.

3. Impact on renal function

(1) To measure creatinine and BUN activity in the serum, blood was collected using the cardiac puncture method after the end of the experiment. Blood was solidified at room temperature for 30 minutes, centrifuged at 3,000 rpm for 15 minutes, and the serum was separated and analyzed by the Department of Clinical Pathology, Daejeon, Korea. The content of creatinine was measured by a biochemical automatic analyzer using the principles of Creatinine Jaffe Method and the content of BUN using the principle of Kinetic UV assay for urea / urea nitrogen

(2) Creatinine ( Cr ) content

The creatinine level was 0.5 ± 0.1 ㎎ / ㎗, 0.6 ± 0.1 ㎎ / ㎗ in control group, 0.5 ± 0.1 ㎎ / ㎗ in GST 200 ㎎ / ㎏ group and 400 ㎎ / ㎏ group showed 0.4 ± 0.1 ㎎ / ㎗ and showed no difference compared to normal group.

(3) Blood urea nitrogen (BUN) content

The mean BUN was 23.3 ± 2.7 ㎎ / ㎗ in the normal group, 20.5 ± 4.9 ㎎ / ㎗ in the control group, 20.4 ± 3.0 ㎎ / ㎗ in the GST 200 ㎎ / ㎏ group and 400 ㎎ / ㎏ group showed 23.0 ± 3.7 ㎎ / ㎗ and showed no difference compared with normal group.

4. Effect on antioxidant efficacy

(1) Antioxidant efficacy measurement

1) Measurement of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability

① Free radical scavenging activity test is a method using stable free radical DPPH. The GST extract was diluted to a final concentration of 1, 10, 100, 1,000 (㎍ / ml), and 150 μl of 0.2 mM DPPH solution dissolved in ethanol and 100 μl of GST extract were mixed to obtain 37 Lt; 0 > C for 30 minutes. After the reaction, the absorbance was measured at 517 nm. The control solution of the sample solution was added with distilled water, and ethanol was added to the DPPH solution as a control. DPPH free radical scavenging rate was calculated according to the following [Table 1].

Erase rate (%) = ( Absorbance of the control group - Absorbance of the sample added group ) × 100 Absorbance of control group

② Effect on DPPH radical scavenging ability

The DPPH scavenging rate of GST extracts was 0.3 ± 1.8% at the concentration of 1 ㎍ / ㎖, 1.7 ± 1.3% at the concentration of 10 ㎍ / ㎖, 4.5 ± 1.0% at the concentration of 100 ㎍ / ㎖ and 1,000 ㎍ / 41.3 ± 1.8%, indicating that the radical scavenging ability was increased in a concentration-dependent manner

(2) Measurement of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging ability

The ABTS assay method was modified for 96 well plates. GST extract was diluted to a final concentration of 1, 10, 100, 1,000 (㎍ / ㎖) and ABTS solution was diluted with 7.4 mM ABTS (2,2azinobis- (3-ethylbenzothiazoline-6-sulfonic acid) And 2.6 mM potassium persulphate were prepared and allowed to stand in a dark place for one day to form a cation (ABTS +). The absorbance was measured at 734 nm and the absorbance was diluted to 1.5 or less. 150 μl of the diluted ABTS + 5 μl each of the extracts were mixed, reacted at room temperature for 10 minutes, and the absorbance was measured at 734 nm. The antioxidant capacity of the control group was determined by using percentage of ABTS radical scavenging ability. (Table 2)

Erase rate (%) = (1- (Absorbance of the sample-added group) ) × 100 Absorbance of control group

② Effect on ABTS radical scavenging ability

The ABTS scavenging rate of GST extract was 1.9 ± 1.0% at the concentration of 1 ㎍ / ㎖, 2.9 ± 1.2% at the concentration of 10 ㎍ / ㎖, 8.1 ± 1.4% at the concentration of 100 ㎍ / ㎖, 69.1 ± 2.5%, indicating that the radical scavenging ability was increased in a concentration-dependent manner

5. Effect on anti-inflammatory

(1) Anti-inflammatory measure

1) Nitric oxide (NO) measurement

① NO concentration was measured by using griess reagent system. RAW 264.7 cells were plated at 1.5 × 10 ⁵ cells / well in a 96-well plate and cultured for 24 hours. After the culture, the cells were replaced with fresh culture medium. GST extracts were treated with 1, 10, 100 (㎍ / ㎖) and 1 ㎍ / ㎖ of LPS, respectively. 50 μl of N1 buffer was added to each well and reacted at room temperature for 10 min. Then, 50 μl of N2 buffer was added to each well and reacted for 10 min. After the reaction, the absorbance was measured at 540 nm. The concentration of NO in the culture medium was determined using the standard curve of concentration of nitrite standard.

② Effect on production of nitric oxide (NO)

The NO production of RAW 264.7 cells was 34.4 ± 2.7% when the control group was 100.0 ± 3.7%, 101.6 ± 14.6% and 10 μg / ㎖ at the concentration of 1 ㎍ / ㎖, respectively (**: p <0.01, *: p <0.05) at the concentration of 100 and 100 ㎍ / ㎖, respectively Lt; / RTI &

2) Measurement of cytokine production

1) Inflammatory cytokines were cultured in a 12-well plate at a density of 2 × 10 ⁵ cells / well in a 12-well plate and replaced with fresh culture medium. GST extracts were added at 1, 10, and 100 μg / ), Treated at a concentration of 1 μg / ml of LPS, and cultured for 24 hours in a cell culture incubator (37 ° C., 5% CO ₂ ). After centrifugation, IL-1β, IL-6 and TNF-α were measured with Luminex as a supernatant.

② Effect on IL-1β production

The amount of IL-1β produced by RAW 264.7 cells was 48.0 ± 6.2 pg / ㎖ in the control group and 12.6 ± 1.0 pg / ㎖ in the normal group, and 43.3 ± 12.8 pg / ml at the concentration of 1 ㎍ / (*: P <0.001) at concentrations of 10 and 100 μg / ml compared to the control group, which was 28.1 ± 4.4 pg / ml at a concentration of 10 μg / ml and 29.2 ± 7.1 pg / 0.05) in the control group.

③ Effect on IL-6 production

The amount of IL-6 produced by RAW 264.7 cells was 1269.3 ± 8.9 pg / ㎖ in the control group, 200.6 ± 71.3 pg / ㎖ in the normal group, and 1169.0 ± 300.3 pg / ml at the concentration of 1 ㎍ / , shown as 10 ㎍ / ㎖ 991.0 ± 12.6 pg / ㎖ in concentration, 935.3 ± 127.2 pg / ㎖ at 100 ㎍ / ㎖ concentration, with significance at a concentration of 10, 100 (㎍ / ㎖) than the control group (*: p < 0.05) in the control group.

④ Influence on TNF-α production

The amount of TNF-α produced by RAW 264.7 cells was 6451.3 ± 557.6 pg / ㎖ in the control group and 573.0 ± 52.3 pg / ㎖ in the normal group, and 6151.8 ± 779.6 pg / ㎖ at the concentration of 1 ㎍ / , 6053.8 ± 187.0 pg / ml at a concentration of 10 μg / ml, and 6041.8 ± 72.5 pg / ml at a concentration of 100 μg / ml.

5 . Effect on skull fracture

(1) Causing fracture of skull and sample treatment

The 11-week-old SD rats were anesthetized with anesthetics (0.5 ml of ketamine + 0.1 ml of rumen) to the 13-week-old rats after 2 weeks of stabilization, and the surrounding skulls were cleaned and fixed to the operating table. The skull of the fixed rat was opened using a mass, and a fracture of 8 ㎜ in diameter was implanted into the skull using Trephine Bur. The experimental group was divided into four groups: normal control group, distilled water control group, GST administration group at 200 ㎎ / ㎏ and GST administration group at 400 ㎎ / ㎏. Were orally administered at a dose of 200 [mu] L each. During the course of the experiment, a free diet was given. The GST dose was calculated based on a single intake of 60 kg of body weight per adult, 310 g of rat body weight, and skull fracture The oral administration was performed for 8 weeks in total, based on the 0th point.

(2) Influence on fusion of skull fracture

1) X-ray filming

To determine the effect of GST on skull fracture union, X-ray was taken at intervals of 4 weeks from 0 to 8 weeks, which caused skull fracture. As a result, in the control group fractures were caused from 0 to 8 weeks of the skull fracture The bone margins were clear and union was not performed.

On the other hand, as GST 200 and 400 ㎎ / ㎏ treated group, the margins were blurred and the outline of the bone became clearer as the experiment proceeded at the 8th week, and the fusion process of the fracture was proceeding (FIG. 1).

2) CT shooting

To confirm the effect of GST on the union of the skull fracture, micro-CT scan after the end of the experiment showed that the union of the induced site of the skull fracture in the control group was 8 ㎜, In GST 200 and 400 ㎎ / ㎏ administration group, bone unification process was faster than control group at 8th week. In particular, it was confirmed that the bone union was rapidly proceeding in the 200 mg / kg administration group as compared with 400 mg / kg (FIG. 2).

3) Bone density measurement

After the end of the experiment, the skulls were analyzed by 3D micro-CT and the bone density was calculated by calculating the ratio to BMD (Bone Mineral Density). As a result, the control group showed 0.59 ± 0.01 ㎎ / ㎠, whereas GST 200 ㎎ / 0.67 ± 0.05 ㎎ / ㎠ for GST and 0.64 ± 0.02 ㎎ / ㎠ for GST 400 ㎎ / ㎏ administration group, respectively (Fig. 3).

4) Bone area Measure

After the end of the experiment, the skulls were analyzed by 3D micro-CT and the bone area was measured by calculating the ratio to the BV (bone volume). As a result, the control group showed a result of 3.67 ± 0.70 U ^ 3, whereas the GST 200 mg / ( P <0.01, *: p <0.05) in the group treated with GST 400 ㎎ / ㎏ and 0.4.99 ± 0.71 U ^ 3 in the group treated with GST 4).

Thus, the present invention provides a tamoxifen preparation having remarkable efficacy for restoring skull fracture without toxicity to liver (kidney) and kidney (kidney kidney).

The present invention is very useful in an industry for producing, processing, distributing, selling, and researching functional foods and medicines for recovering and treating skull fractures using herbal medicines and herbal medicines.

Claims

Ginseng, white liquor, gyeongryeong, licorice, sorghum, peony, celestial ginseng, angelica, hwanggi, broiler, natural copper,
80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of licorice, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, 80 to 120 parts by weight of ginseng, And 120 to 180 parts by weight of bone marrow, 120 to 180 parts by weight of wild birds, 80 to 120 parts by weight of wild birds, 80 to 120 parts by weight of birds, 120 to 180 parts by weight of natural copper, (I).

delete

A pharmaceutical composition comprising the dermal decontamination composition for improving skull fracture according to claim 1.