KR100284657B1 - Osteoporosis treatment using Astragalus extract as an active ingredient - Google Patents

Abstract

The present invention relates to the use of Astragali Radix extract as a therapeutic agent for osteoporosis. Specifically, the present invention relates to a therapeutic agent for osteoporosis comprising the extract of sulfuric acid, a natural herbal medicine as a lower alcohol, adding water and then partially purified with hexane, as an active ingredient, which is caused by various causes such as aging or menopause. It can be effectively used to prevent and treat osteoporosis, which has no side effects.

Description

Osteoporosis treatment using Astragalus extract as an active ingredient

The present invention relates to the use of Astragali Radix extract as a therapeutic agent for osteoporosis.

More specifically, the present invention relates to a therapeutic agent for osteoporosis, which comprises the extract of sulfur, a natural medicinal herb, as a lower alcohol and fractionated with n-hexane, as an active ingredient, which is caused by various causes such as aging or menopause. It can be effectively used to prevent and treat osteoporosis without side effects.

Bone plays a role in preserving bone mass and structure necessary as a physical support of the body, and plays an important role in maintaining blood concentrations of calcium, such as calcium (Ca ²⁺ ) storage. In order to perform this function, the bone always needs to be in harmony with the decomposition and remodeling action, and in this process, the bone absorption and the bone formation are in a dynamic state. When such an equilibrium relationship between bone absorption and bone formation is broken, bone absorption may be relatively higher than bone formation, resulting in osteoporosis in which bone density or amount of bone is reduced and bone strength cannot be maintained.

Osteoporosis is a disease in which the amount of bone is reduced overall and the bone is structurally weakened, so that fractures and morphological deformations are easily caused by small impacts. The patient first shows a decrease in the space and cortex thickness occupied by the bone source (Reilly , DT and Burstein, AH, 1994, J. Bone Joint Surg., 56, 1001-1022).

Osteoporosis can be divided into various types according to its type and cause, including the type I osteoporosis caused by menopause and the type II osteoporosis caused by aging (Dempster. DW and Lindsay, R., 1993, Lancet, 341, 797-801). Although many studies have not been conducted on the causes of osteoporosis, type I and type II osteoporosis are caused by endocrine disorders in many cases and in addition to calcium, phosphate and vitamin D due to reduced absorption of diseases, malnutrition or calcium ions. It is known to result from a deficiency (Sambrook, P. et al., 1993, N. Engl. J. Med., 328, 1747-1752; Marie, PJ et al., 1989, J. Clin. Endocrinol. Metab. , 69, 272-279; Krane, SM and Holick, MF, 1994, in Harrison's Principles of Internal Medicine, vol. 2, pp 2172-2183).

In addition, in osteoporosis patients, a decrease in the concentration of IGF-1 (Insulin-like growth factor) in plasma is observed, which correlates with Bone Mineral Density (BMD) and is a hormone important for muscle metabolism (thyroids). And parathyroid PTH, cortisol and estrogen, and vitamin D3 (Johansson, AG et al., 1992, J. Int. Med., 232, 447-452; Abbasi). , AA et al., 1993, J. Am. Genetrics Soc., 41, 975-982). In addition, much research is currently being conducted on the relationship between hormonal changes and bone density.

Since Type I and Type II osteoporosis are diseases that occur over decades, it is not enough to take some simple drugs for a short time to treat them. Until now, estrogen or testosterone has been generally used as a therapeutic agent for osteoporosis, and calcium, phosphate, fluoride, ipriflavone, vitamin D, and calcitonin have been reported to be used as treatments. In case of greater side effects, it is necessary to develop a new osteoporosis treatment that has no side effects.

In addition, until now, the osteoporosis treatment agent that inhibits excessive bone resorption by inhibiting the function of osteoclasts has been mainly developed. However, suppressing bone resorption alone makes it difficult to recover the risk of fracture due to the reduction of bone mass. Drugs should be developed to facilitate this.

On the other hand, osteoporosis is a chronic disease, so it is necessary to develop a number of easy test methods in a preventive way. Since bone tissue is a hardened petrified tissue, there are limitations in the method of analysis. However, the following methods have been developed as an initial diagnosis of osteoporosis. Specifically, single dual-photon absorptiometry, quantitative computed tomography, X-ray based dual energy densitometry, and neutron activity analysis of total calcium Methods (neutron activation analysis), etc. (Joffe, I. and Epstein, S., 1991, Seminars Arthr. Rheum., 20 (4), 256-272). However, only one measurement of bone density does not determine the deformation of the bone, there is a disadvantage that can be known to decrease the bone mass only measured at intervals of time over several consecutive years.

Another method to diagnose osteoporosis that is accurate and sensitive but requires a lot of skill is histology of bone deformation using hematoxylin and eosin staining (Cameron, JR and Sorenson, J., 1963, Science, 142, 230). In addition, blood or urine tests can be diagnosed indirectly. Physiological changes can be examined through various biochemical clinical tests that examine the distribution of cells in the blood and quantitative analysis of hormones.

Astragali Radix (Astragalus membranaceus Bunge) is a herb that belongs to the legume (Leguminosae), and according to the explanation of Ishijin of the New Farm, the color is yellow and is named after the long term to protect the human body. In Korea, Astragalus roots have been used a lot as a medicine after ginseng in the past, and Astragalus has been used as a tonic, diuretic, and antiperspirant. In particular, oriental medicine is used for hundreds of prescriptions such as Bogiikgitang, Hwanggigunjungtang, Sipjeondaebotang, Kamimidaebotang, Palbohoechuntang, and Cheongseoikgitang.

In addition to Korea, Astragalus, China (A. chinensis L .; A complanatus R. Br .; A. sinicus; A. chrysopterus), Inner Mongolia (A. membranaceus var. Mongholicus), Japan (A. aitosensis; Hedysarum iwawoki Hara; A. It is distributed in adsurgens Pallas Subsp.fujisanensis Kita; A. polybotrys Hands-Mazz.), Egypt (A. spinosus Vahl; A. trigonus), Bulgaria (A. onobrychis), etc. It is known that a lot is produced.

To date, triterpenoid glycoside strains and isoflavonoid strains among flavonoids have been mainly reported as components. Specifically, polysaccharides isolated from Astragalus have been reported to improve the function of T cells in cancer patients, and triterpenoids and flavonoids are known to have antioxidant, anti-inflammatory and blood pressure lowering activities (Huaping , L., Yau, ZO and Bo, G., 1994, Chin.H. Plast.Surg. And Burns, 3 (102), 138-141; Geng, CS, 1986, Chin.J. Mod. Dev.Trad Med., 6 (1), 62-64); Zhang, Y. D. et al., 1992, Acta Pharm. Sin., 27 (6), 401-406; Harborne, J. B. and Baxter, H., 1993, Phytochemistry Dictionary, Taylor & Francis, London, Washington DC., Pp 673; Tang, W. and Eisenbrand, G., 1992, Chinese Drugs of Plant Origin, Springer-Verlag, Berlin, Heidelberg, New York, pp. 191).

In addition, the present inventors have already described the 2-benzendicarboxylic acid diisononylester, β-sitosterol, 3, in the fraction obtained by partially purifying the alcohol extract of the Astragalus root with n-hexane. -o-β-D-galactopyranosyl-β-sitosterol (3-o-β-D-galactopyranosyl-β-sitosterol), stigmas-4-en-6β-ol-3-one (stigmast-4- en-6β-ol-3-one), 7-hydroxy-4'-methoxy-isoflavone and 9Z, 12Z-octadecadienoic acid (9Z, 12Z octadecadienoic acid) (Kim, JS and Kim, CS, 1997, Kor. J. Pharmacogn., 28 (2): 75-79; Kim, JS et al., 1996, Kor. J. Pharmacogn., 27 (4): 336-341).

In particular, among the above components, 7-hydroxy-4'-methoxy-isoflavone (formononetin; formononetin) and stigmas-4-ene-6β-ol-3-one, and the organic extracts containing them are growth hormones It has been reported to increase the production of growth hormone (GH) by acting as a secretory stimulating factor. Growth hormone is secreted by the animal's pituitary gland, which inhibits protein loss and delays physiological aging in older adults or the elderly, which can be used for a variety of diseases. The weight of the increase, etc. can prevent diseases caused by aging. In addition, growth hormone has been reported to increase the weight and size of the liver and improve protein synthesis in terminal tissues such as muscle, bone and connective tissue (Kaiser, FE et al., 1991, J. Am. Geriatr. Soc , 39, 235-240; Marcus, R. et al., 1990, J. Clin. Endocrinol. Getab., 70, 519-527).

Therefore, the present inventors continue to search for a substance that prevents or treats osteoporosis from various traditional Chinese medicines. As a result, fractions of Astragali Radix extracted with ethanol and partially purified by n-hexane and components included therein The present invention was completed by morphometrically confirming that they can be used for the treatment of type I and type II osteoporosis by increasing the area of bone trabecular.

It is an object of the present invention to provide a pharmaceutical composition comprising Astragali Radix extract as an active ingredient as an agent for preventing and treating osteoporosis.

In order to achieve the above object, the present invention provides a pharmaceutical composition for treating osteoporosis, comprising as an active ingredient the extract of Astragali Radix with a lower alcohol and partially purified by an organic solvent.

At this time, the Astragalus extract is preferably prepared using ethanol as the lower alcohol, n-hexane, ethyl acetate or n-butanol as the organic solvent, 7-hydroxy-4'-methoxy-isoflavone and its Derivatives or stigmas-4-ene-6β-ol-3-ones and derivatives thereof and the like. Osteoporosis also includes Type I osteoporosis and Type II osteoporosis and the like.

Hereinafter, the present invention will be described in detail.

The present invention provides a use of a pharmaceutical composition comprising as an active ingredient Astragali Radix extract, a compound purified therefrom, and derivatives thereof, as an active ingredient, for the prevention and treatment of osteoporosis.

The present invention extracts sulfuric acid with a lower alcohol, suspended with distilled water and then fractionated again by adding an organic solvent to prepare a sulfuric acid extract. At this time, it is preferable to use ethanol as the lower alcohol, and it is preferable to use n-hexane, ethyl acetate, n-butanol and the like as the organic solvent. In particular, it is more preferable to use n-hexane as the organic solvent.

In addition, the sulfur group is extracted with a lower alcohol, suspended with distilled water and then extracted by adding n-hexane as an organic solvent and then repeated by chromatography to carry out 7-hydroxy-4'-methoxy-isoflavone of the formula (1) Or stigmas-4-en-6β-ol-3-one of formula (2).

For reference, the Astragalus extract and 7-hydroxy-4'-methoxy-isoflavone and stigmas-4-ene-6β-ol-3-one have been previously described by the present inventors as a growth hormone secretion stimulating factor. It has been confirmed.

The present invention provides a pharmaceutical composition comprising 7-hydroxy-4'-methoxy-isoflavone and derivatives thereof or Stigmas-4-en-6β-ol-3-one and derivatives thereof as an active ingredient for the prevention of osteoporosis. And therapeutic agents.

In order to investigate the osteoporosis treatment effects of the Astragalus extract and the compounds contained therein, the ovaries were extracted from the animal models having type I and type II osteoporosis, respectively, using rats and gene mutations that became type I animal models. Animals developed as Type II animal models that promote aging and osteoporosis are selected and tested.

In general, there are various methods for studying the therapeutic and prophylactic effects of osteoporosis such as biochemical, morphological, and physiological research methods, but the most easily accessible is morphological method. In addition, morphological studies can be divided into histopathological analysis, morphometric analysis, and immunohistochemical analysis by optical microscope.

The present invention investigates the improvement of osteoporosis by measuring bone density by analyzing loss of bone trabecualar in experimental animals by morphometric measurement using an image analyzer. As a result, as shown in Table 4, the Astragalus extract is significantly effective for type I and type II osteoporosis and preferably for type I.

Osteoporosis is a pathophysiological disease in which many complex processes occur at the same time. Therefore, it is difficult to analyze the pathogenesis and current state as one indicator. Therefore, it is necessary to evaluate it using various indicators. Therefore, the present invention is to investigate the change in erythrocytes, hemoglobin, hematogrit and the like by the total blood cell test method when administering astragalus extract or the like to the ovarian extract animal and the weight change of the animal. As a result, it was confirmed that there was no significant change in total blood cell count and body weight due to the administration of Astragalus extract.

Therefore, it can be seen that the pharmaceutical composition comprising the Astragalus extract of the present invention as an active ingredient is very useful as an agent for preventing and treating osteoporosis.

In order to use the compound of the present invention prepared by the above process as a therapeutic agent, it may be prepared by a known method in the pharmaceutical field, and may be a carrier or excipient (pharmaceutically), itself or pharmaceutically acceptable. It may be prepared and used in the form of a powder, granules, tablets, capsules or injections by mixing with diluents and the like.

The human dose of the active ingredient according to the present invention is appropriately selected according to the absorption of the active ingredient in the body, the rate of water activation and excretion, the age, sex and condition of the patient, the severity of the disease to be treated, but generally 1 30-200 mg per day is recommended, but 50-150 mg is preferred. Therefore, the unit dosage form of the present invention is prepared to have an amount of 20-50 mg of the active substance of the present invention in consideration of the above-mentioned effective amount range. Preferably it is formulated to contain 10-30 mg. The unit dosage form thus formulated may be administered several times at regular intervals or by using a specialized dosing method according to the judgment of an expert who monitors or observes the administration of the drug as needed and the needs of the individual. It may be administered once to three times a day.

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

The following examples illustrate the invention, but the scope of the invention is not limited by the examples.

Example 1 Preparation of Astragalus Extract

Astragalus roots grown in Sindong-eup, Jeongseon-gun, Gangwon-do, South Korea were directly purchased and evaluated. Powdered yellow roots were extracted and 80% aqueous ethanol was added and extracted four times at room temperature one week apart (A). The ethanol extract was concentrated under reduced pressure, dried, and distilled water was added thereto, followed by systematic separation in the order of n-hexane (B), ethyl acetate (C), and n-butanol (D), and these were divided into a partition layer and a water layer (E), respectively. Distilled water was added to the residue, and the mixture was left at room temperature for 2 days and then extracted again. Another method was extracted for 2.5 hours at 90 ℃ with water cut finely cut off the Astragalus root (F). Double n-hexane fractions (40 g) were subjected to primary silica gel column chromatography (fixed phase: silica gel 60 (Merck No. 7734); size 1300 * 5.0 cm; mobile phase: chloroform: acetone = 9: 1 → 1: 1 → 100% Ethanol; flow rate 3.8 ml / min) and pre-separated into 25 subfractions. The 15th subfraction was subjected to secondary silica gel column chromatography (fixed phase: silica gel 60 (Merck No. 9385); size 63.0 * 2.0 cm; mobile phase: chloroform: methanol = 9: 1; flow rate 3.0 ml / min → 1.0 ml / Min → 3.0 ml / min) to obtain a somewhat purified fractions and collected and concentrated under reduced pressure at about 40 ℃. When this was stored at 4 ° C, white crystals precipitated. This was filtered under reduced pressure to separate the mother liquor from the yellow liquor, and the white crystals were still yellow. Therefore, the resultant column was still yellow. Third column chromatography (fixed phase: silica gel 60 (Merck No. 9385); : Methanol = 96: 4) was further performed. From this can be recognized by absorbance at 254 nm and 356 nm and placed on a TLC plate (mobile phase: methylene chloride: ethyl acetate = 7: 3) sprayed with anisealdehyde-sulfuric acid reagent and not heated to 7-hydroxy- 4'-methoxy-isoflavone can be obtained.

<Example 2> Preparation of Astragalus Extract

The n-hexane fraction (40 g) of the sulfur group obtained by the same procedure as in Example 1 was subjected to primary silica gel column chromatography (fixed phase: silicic acid 60 (Merck No. 7734); size 1300 * 5.0 cm; mobile phase: chloroform: acetone = 9: 1 → 1: 1 → 100% ethanol; flow rate 3.8 ml / min) and separated into 25 small fractions. The 15th subfraction was subjected to secondary silica gel column chromatography (fixed phase: silica gel 60 (Merck No. 9385); size 63.0 * 2.0 cm; mobile phase: chloroform: methanol = 9: 1; flow rate 3.0 ml / min → 1.0 ml /Min→3.0 ml / min) to obtain a somewhat purified fraction, which was collected and concentrated under reduced pressure in a water bath of about 40 ℃. To the mixture was added 2-3 drops of chloroform, which were dissolved and stored at room temperature and 4 ° C for 24 hours to precipitate white crystals. This was washed with methanol to separate the mother liquor from the yellow liquid, and the white crystals were dissolved in chloroform to repeat the above process to obtain an amorphous powder. It is placed on a TLC plate (mobile phase: methylene chloride: ethyl acetate = 7: 3) and sprayed with an anisealdehyde-sulfuric acid reagent and heated to turn yellow, then turn into scarlet. -All-3-one was obtained.

Reference Example 1 Analysis of Growth Hormone Secretion Stimulation by Using Astragalus Extract

In the present invention, each of the Astragalus extract fractions obtained in Example 1 was examined for growth hormone secretion stimulatory activity to confirm the characteristics of each fraction. To investigate the activity, rat pituitary cells were aliquoted at a concentration of 1.5 × 10 cells / ml in 24-well plates and incubated for 5 days. Next, 100 μl of each fraction was dissolved in 1 ml of dimethylsulfooxide (DMSO), added to each well, incubated for 15 minutes, and the growth hormone concentration in the medium was measured by a double antibody RIA method (Tanaka, T. et al., 1983, J. Clin.Endocrinol., 56 (1), 18-20; Wroblewski, VJ et al., 1991, Endocrinology, 129 (1), 465-474). At this time, the standard curve for growth hormone was examined at a concentration ranging from 10 ng / ml to 121 pg / ml, and rat growth hormone including ¹²⁵ I labeled growth hormone, rat growth hormone monkey antiserum, and rat growth hormone as a kit. Assay kits (NIDDK, Harbor-UCLA Medical Center, CA, USA) were used. Next, monkey antibody IgG (CAPPEL) to goat antibody IgG and polyethylene glycol (PEG) at a final concentration of 6% were added as a secondary antibody to precipitate an immunocomplex and measure radioactivity contained in the precipitate.

The activity to secrete growth hormone secretion of each fraction of the Astragalus extract investigated by the above process is shown in Table 1 as the radioactivity contained in the growth hormone, from which fraction B, fraction C and fraction D is Bonferonni multiple comparison method (Bonferonni multiple In comparison, P <0.01), the growth hormone secretion stimulation activity was superior to the reference.

Growth Hormone Secretion Stimulation Activity Using Astragalus Extract Fraction Concentration (mg / ml) Growth hormone (ng / ml: mean ± SE; n = 4) standard - 4.75 ± 0.55 A 2.7 6.19 ± 0.98 B 33.5 119.40 ± 27.16 C 21.5 107.90 ± 28.23 D 9.3 112.86 ± 7.39 E 3.8 18.22 ± 11.48 F - 4.52 ± 0.62

Example 3 Screening of Animals for Investigation of Osteoporosis Treatment Effect of Astragalus Extract

The present invention is a type I and type II osteoporosis model among the various animal models that cause osteoporosis. SAM P6 mice were selected to test the effect of Astragalus extract.

Specifically, in the type I osteoporosis animals, Sprague-Dawley rats weighing 250-300 g at 10 weeks of age received from the Korea Research Institute of Chemistry (Daejeon) underwent surgery to remove bilateral ovaries under general anesthesia and aseptic manipulation. Then used. At this time, the comparison group (Sham group) was used to undergo the above surgical procedure except for ovarian extraction, and these animals were administered for 9 weeks from 11 weeks to 20 weeks after the recovery period of 1 week. In addition, type II osteoporosis animals used a SAM P6 mutant strain that promotes aging and osteoporosis among SAM mouse models developed by Takeda et al. This is known to decrease bone density from 10 weeks of age, and the comparison group used SAM R1 mice (Kim Jung-sook et al., 1994, The Korean Society for Applied Drugs, 5, 23-35; Takeda, T. et al., 1994 , in The SAM model of senescence, pp 15-22, Elsevier Science BV; Suda, T., 1994, in The SAM model of senescence, Takeda, T. (ed), pp 47-52, Elsevier Science BV).

<Example 4> Dosing effect investigation of Astragalus extract

The present invention, in order to investigate the osteoporosis treatment effect of the Astragalus extract, extract extracted with 80% aqueous ethanol, concentrated under reduced pressure and dried under reduced pressure to the experimental animals selected in Example 3 and then partially purified by n-hexane again Was continuously administered and the changes in bone strain and body weight change of the animals were measured in the following Examples. Specifically, animals that induce type I osteoporosis are divided into a control group and an administration group.In the case of type I, the control group (n = 11) receives a 10% Tween 80 solution corresponding to 1 mg / kg, and the administration group has an astragalus extract (n = 10). ) Was intraperitoneally injected with a solution of 10% Tween 80 at a dose of 1 mg / kg once daily for 9 weeks, followed by blood collection at 0, 7, 14, 21 and 30 days for total blood count (CBC test). , complete blood cell counts (RBC, HGB, HCT) were performed and after 9 weeks (20 weeks of age), tibias were separated and stored in 4% formalin solution (10% dilution) to determine bone mass.

As a result, as shown in Table 2, the preoperative average body weights of the Type I control group and the administration group were 261.28 ± 6.95 g and 243.67 ± 10.48 g. After 9 weeks, the control group was 388.24 ± 26.37 g and the administration group was 379.72 ± 20.5 g. As the experiment continued, there was no significant difference in body weight between the two groups.

Dosing period Control Administration group 10 (before surgery) 261.28 ± 6.95 243.67 ± 10.48 11 (before dose) 275.25 ± 10.16 259.60 ± 7.62 12 (1 week of administration) 291.70 ± 13.33 273.19 ± 8.89 14 (3 weeks) 339.41 ± 17.17 312.97 ± 17.03 16 (dose 5 weeks) 343.74 ± 21.68 339.78 ± 17.06 18 (dose 9 weeks) 366.96 ± 23.39 351.92 ± 18.18

Example 5 Total Blood Cell Test (CBC Test)

In order to investigate the effect of the Astragalus extract in the experimental animals, a total blood cell test was performed. Osteoporosis is a pathological and physiological disease in which many complex processes occur simultaneously. Therefore, as one indicator, it is difficult to analyze the mechanism and current state of the disease. Therefore, it should be evaluated using various indicators. Specifically, leukocytes (WBC), lymphocytes (lymphocytes, LY), monocytes (monocytes, MO), granulocytes (GR), Levels of red blood cells (RBC), hemoglobin (Hb) and hematocrit (Hmato) were measured. As a result, the amount of erythrocytes, hemoglobin, hematogrit, and the like changed by administration of Astragalus extract in ovarian isolated mice with type I osteoporosis is shown in Table 3.

Preoperative Control group (after 9 weeks of administration) Administration group (after 9 weeks of administration) leukocyte 13.800 ± 1.821 9.760 ± 1.931 15.660 ± 1.840 Lymphocytes 10.883 ± 1.684 7.260 ± 1.616 11.940 ± 1.495 Monocytes 0.567 ± 0.092 0.900 ± 0.084 0.950 ± 0.155 Granulocyte 1.750 ± 0.363 1.620 ± 0.297 2.125 ± 0.485 Red blood cells 6.986 ± 0.369 7.194 ± 0.212 7.434 ± 0.213 hemoglobin 14.744 ± 0.324 14.720 ± 0.361 14.960 ± 0.183 Hematocrit 40.888 ± 2.221 40.420 ± 1.665 42.780 ± 0.455

<Example 6> Density measurement of bone

In the present invention, bone density was measured in the experimental animals to investigate the osteoporosis treatment effect of the Astragalus extract. Specifically, in the present invention, the loss of bone alcohol was analyzed by the morphometric method using an image analyzer (Vidas image analysis system, Zeiss Co., Germany) (Cressent, M. et al., 1981, Proc. Soc. Exper. Biol. Med., 166, 92-95; Durbridge, TC et al., 1990, Calcified tissue internat., 47, 383-387; Mann, DR et al., 1990, J. Clin.Endocrinol. Metab., 71, 105-110).

(3-1) organization treatment

For histopathological observation of bone, bone tissue was harvested and fixed in 10% formalin solution and left to demineralized solution (De-cal rapid solution, National diagnostics, U.S.A.) for 1-2 days. Next, the area to be observed in the bone tissue was cut with a surgical knife, followed by stepwise dehydration from 70% to 100% alcohol and acetone. Bone tissue embedded in paraffin was cut to 5 μm with a microtome and stained with hematoxylin eosin (H & E).

(3-2) tissue image analysis

The degree of loss of bone trabecular due to osteoporosis was directly analyzed using a morphometric method using an image analyzer. Specifically, the image analyzer was digitalized and the area of each bone strain was quantitatively calculated automatically on the computer screen along the outline. Specifically, in the proximal part of the tibia, the individual areas of all bone fragments in the rectangle with a reference area of 0.196304 × 10 ⁿ μm ² whose horizontal side length is about 2/3 of the length of the growth plate in the proximal part of the tibia were obtained, and then The total area of the bone strains was determined by multiplying the number.

The measurement results are shown in Table 4. The results were analyzed using the ^Systoma® program (SYSTAT inc., Evanston, Ill, USA), which was defined as meaning less than P <0.05 in Bonferroni multiple comparison analysis. . The results were found to be significantly different in both the Fisher method in addition to the Bonferroni method.

Control group (unit: × 10 ⁵ μm ² ) Administration group (Unit: × 10 ⁵ μm ² ) 1 (11 objects) 290914.15 ± 39592.87 505310.72 ± 73989.07 2 (10 objects) 314159.23 ± 35433.63 539677.56 ± 72438.73

As described above, the Astragalus extract of the present invention effectively increases only bone density without a change in body weight and has no side effects as a natural herbal medicine, so that the pharmaceutical composition having the active ingredient thereof is very useful as an agent for preventing and treating osteoporosis. Specifically, osteoporosis caused by various causes such as aging or menopause can be treated with minimal side effects due to long-term administration, and thus it is expected to be widely used in the future.

Claims (4)

A therapeutic agent for osteoporosis comprising Astragali Radix as an active ingredient, which is extracted with a lower alcohol and partially purified by an organic solvent. According to claim 1, Astragalus extract is prepared using ethanol as a lower alcohol, n-hexane, ethyl acetate or n-butanol as an organic solvent, preferably n-hexane as an organic solvent. Osteoporosis treatment agent. 3. The extract according to claim 1, wherein the Astragalus extract comprises 7-hydroxy-4′-methoxy-isoflavone and derivatives thereof or stigmas-4-en-6β-ol-3-one and derivatives thereof. An agent for treating osteoporosis, characterized in that. The therapeutic agent for osteoporosis according to claim 1, wherein the osteoporosis comprises type I osteoporosis and type II osteoporosis.