KR20090093646A - Composition comprising the mixed extract of Carthami Semen, Rehmanninae Rhizoma Preparat and Amomi Fructus for preventing and treating osteoporosis - Google Patents

Abstract

A pharmaceutical composition for preventing and treating osteoporosis, which contains mixed herbal extract of Carthami Semen, Rehmanninae Rhizoma Preparat, and Amomi fructus is provided to increase the viability of MG-63 cell which is osteoblast cell line, MG-63 cell and increase estradiol and osteocalcin. A pharmaceutical composition for preventing and treating osteoporosis comprises mixed herbal extract of Carthami Semen, Rehmanninae Rhizoma Preparat and Amomi fructus as an active ingredient. The mixed herbal extract is isolated using water, low alcohol of 1-4 carbon atoms or their mixture solvent. The mixed herbal extract of Carthami Semen, Rehmanninae Rhizoma Preparat and Amomi fructus is contained in the weight ratio of 1-10:1-5:1-5.

Description

Composition comprising the mixed extract of Carthami Semen, Rehmanninae Rhizoma Preparat and Amomi Fructus for preventing and treating osteoporosis}

The present invention relates to a pharmaceutical composition for the prevention and treatment of osteoporosis diseases, and a dietary supplement for the prevention and improvement, containing a mixed herbal extract of safflower, succinate and sine as an active ingredient.

Roodman GD et al., Advances in bone biology: the osteoclast, Endocrinol Rev. 17 , 308-332, 1996

Document 2 Avioli LV. et al., The osteoporotic syndrome; Detection, prevention & treatment. 3rd ed. Wielis, 1993

[Ref. 3] Kwon, Jae-Hee et al., Bone Mineral Density in Postmenopausal Women: Comparison between Natural Menopause Group and Surgical Menopause Group, Korean Society of Obstetrics and Gynecology, 2000

[Ref. 4] Osteoporosis (osteoporosis), The Korean Society of Bone Metabolism, The Latest Medical History, p17-63, 1991

[Document 5] Seoil Il et al., Intuitive Herbology, Daegu Haany University Press, p295, 2004,

[6] Seoil Il et al., Intuitive Herbology, Daegu Haany University Press, p435, 2004

[Document 7] Seoil Il et al., Intuitive Herbology, Daegu Haany University Press, p174, 2004

[Reference 8] Oh K. et al ., Effect of Rehmaniae glutinosa Libosh extracts on bone metabolism, Clinica Chimaica Acta. 334, 185-195, 2003

Document 9 Dontas I. et al . Protective effect of plant extract from Onobrychis ebenoides on ovariectomy-induced bone loss in rats. Maturitas. 53, 234-242, 2006

Document 10 Suzuki N. et al . , A possible role of estrone produced in adipose tissues in modulating postmenopausal bone density, Maturitas. 22, 9-12, 1995

Srivastava AK, et. al ., Development and application of a serum C-telopeptide and osteocalcin assay to measure bone turnover in an ovariectomized rat model, Calcif Tissue Int. 66 (6), 435-442, 2000

Osteoporosis is a decrease in bone matrix, so the mass of the bone itself is very low, and many small holes, such as rough pumice or sponge, are broken and soft and easily broken. Osteoporosis is divided into primary and secondary osteoporosis. Primary osteoporosis is classified into postmenopausal type 1 and senile osteoporosis. Type 1 osteoporosis is a major cause of post-menopausal estrogen deficiency, which leads to increased bone resorption, resulting in higher calcium in the blood and lower parathyroid hormone secretion, resulting in lower intestinal calcium absorption. It occurs mainly in older women. The mechanism of development is the production of cytokines such as IL-1 (Interleukin-1) and IL-6 in osteoblasts due to estrogen deficiency, and bone loss occurs by activating osteoclasts. In this case, bone loss occurs mainly in the bone, so fractures occur frequently in the spine and wrist. Type 2 osteoporosis is osteoporosis that occurs in men and women aged 65 and older. The mechanism of development is the reduction of 1, 25 (OH) ₂ D ₃ production in the kidneys, which leads to decreased intestinal calcium absorption and increased parathyroid hormone secretion, resulting in loss of cortical bone. And osteoporosis due to increased osteoblast formation (Roodman GD et al., Advances in bone biology: the osteoclast, Endocrinol Rev. 17, 308-332, 1996). In addition to the normal physiological bone loss caused by type 1 and type 2, most of the osteoporosis patients are diseases caused by a combination of genetic factors, environmental factors such as wrong lifestyle, and hormone mismatch. Secondary osteoporosis, on the other hand, refers to osteoporosis caused by other diseases or drugs, and pathological causes include osteoporosis caused by various endocrine diseases (Avioli LV. Et al., The osteoporotic syndrome; Detection, prevention & treatment. ed.Welis, 1993)

Bone metabolism consists of repetition of bone absorption and bone formation, and bone mass is determined by these two processes. However, when menopausal deficiency occurs due to menopause, osteoblasts and osteoclasts, which are involved in bone remodeling, directly and indirectly affect bone loss, causing bone loss. Increased bone resorption compared to bone formation causes osteoporosis, which increases the risk of fracture

There are no clear causes of osteoporosis, but increased number of deliveries, decreased activity, estradiol deficiency, vitamin D deficiency, hormonal disorders, fluoride deficiency, reflex dystrophy, genetic causes, senile Causes, lack of calcium intake or decreased absorption in the intestine. These factors act alone or in combination to induce osteoporosis by enhancing bone resorption or weakening bone formation. ).

Fractures caused by osteoporosis are common in the elderly after 65 years of age and are caused by traumatic mild trauma. Fractures occur mainly in the cervical and electronic regions of the femur. Unlike fractures, the morbidity and mortality rate is high and difficult to treat. In addition, the mortality rate within 6 months after the fracture reaches about 30%, and even if it is recovered, it is difficult to recover to the state before the fracture. According to the report, the incidence of fractures is about 15% in elderly people older than 60 years. Of these, about 40% are fractures caused by osteoporosis, whereas in 60-70 years old women, 60% of fractures are fractures caused by osteoporosis. In women with reduced bone mineral density, the risk of fracture is up to two times higher than in women with normal bone mineral density (Korean Society of Bone Metabolism, Osteoporosis, Medical History, p17-63, 1991). In addition, according to the Health Insurance Review & Assessment Service in the first half of 2000, femur fractures accounted for the highest proportion among Korean hospitalized patients. The risk of dying from osteoporosis is 30%, more than three times that of breast cancer, and is emerging as an important health problem not only in Korea but in many developed countries.

Current treatments for osteoporosis require changes in calcium, vitamin D intake, exercise and lifestyle changes. Medications include estradiol, calcitonin, bisphosphonates, etc. And bone stimulators include fluoride, parathyroid hormone, and the like. However, with the exception of some drugs, nothing is known as a breakthrough yet. Therefore, there is an urgent need for drug treatments that primarily inhibit loss of bone density and secondaryly inhibit bone resorption.

Honghwaja (Carthami Semen) is to dry the flowers and seeds of safflower (Carthamus tinctorius L.) in the Asteraceae, the efficacy of safflower (紅花) and was almost the same, in addition to the active hyeolsan Results (活血散結) and hwalhyeol detoxification (活血解毒It has been shown to be effective in the treatment of changchang and changchang (seobuil et al., Plain herbology, Daegu Haany University Press, p295, 2004,).

Rehmanninae Rhizoma Preparat is the root and stem of the perennial grass of Rehmannia glutinosa (Gaertner) Libosch. Or R. glutinosa Libosch. Var. Hueichingensis (Chao et Schih) It is steamed and dry, but tastes sweet and bitter and warm. It replenishes blood and replenishes jeong (精: basic substance for life and activity). It is effective in treating dizziness and blackening the head. However, Sukji-hwang is easy to purify the wet sand because of its discoloration, and it is easy to refine the wet sand, and the combination of sine reduces viscosity and avoids gastrointestinal disorders. et al., Intuitive Herbology, Daegu Haany University Press, p435, 2004).

Amomi Fructus is a dried fruit of the perennial herb of Amomum villosum LOUR. And A. longiligulare TLWU. Or A. Xanthioides Wall. It is collected when the fruit matures, or dried at low temperatures. Sine is hot and weak in flavor, and has aroma. The spicy taste is acidic, the warm nature is intact, and the aroma is hygroscopic. (I) applies to all symptoms which are coarse or gastric in gastric and spleen. In addition, the cause of the sign is that it has the effect of hydration and gastrointestinal nourishment, and the gastrointestinal tract is inconvenient due to the gastrointestinal tract and vomiting and diarrhea due to indigestion. It has the effect of treating seafaring regurgitation, and it treats thoracic swelling pain. , Gestation, gestation, etc. (Western Il et al., Intuitive Herbology, Daegu Haany University Press, p174, 2004).

However, none of the above documents teaches or discloses that a mixed extract of safflower, sujikuhwang and sinus can be used as a composition for preventing and treating osteoporosis diseases.

Accordingly, the present inventors have conducted research to develop an effective therapeutic agent for osteoporosis, and the mixed herbal extract of safflower, succinate and sine increases the survival rate of osteoblast-like cells, osteoblast-like cells, as well as causes of osteoporosis-induced osteoporosis. The present invention was completed by confirming the effect of increasing the reduced bone (vaginal) amount, estradiol and osteocalcin (phosphorus).

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of osteoporosis disease containing a mixed herbal extract of safflower (Carthami Semen), Sukjiwang (Rehmanninae Rhizoma Preparat) and sine (Amomi Fructus) as an active ingredient .

The present invention also provides a health functional food for the prevention and improvement of osteoporosis disease containing a mixed herbal extract of safflower (Carthamin Semen), Sukjihwang (Rehmanninae Rhizoma Preparat) and Sine (Amomi Fructus) as an active ingredient.

The mixed herbal extract as defined herein includes an extract available in a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, preferably an extract available in water.

Preferred weight blending ratios of the mixed herbals defined herein include safflower (1-10): sucrose sulfur (1-5): sinusoids (1-5), and more preferably safflower (1-6). : Sukji sulfur (1 to 3): characterized in that it comprises the weight ratio (w / w) of the sign (1 to 3).

Osteoporosis diseases as defined herein include type 1 osteoporosis (postmenopausal osteoporosis), type 2 osteoporosis (senile osteoporosis), idiopathic osteoporosis, endocrine and osteoporosis, digestive and osteoporosis, malignant and osteoporosis, drinking and osteoporosis, smoking And osteoporosis and one or more diseases selected from the group consisting of drugs and osteoporosis, preferably type 1 osteoporosis (postmenopausal osteoporosis).

Hereinafter, the present invention will be described in detail.

The mixed herbal extract of the present invention can be obtained as follows.

 The mixed herbal extract of the present invention washes the dried herbal ingredients crushed safflower, succinate sulfur and sine with water to a volume of water of about 1 to 50 times, preferably about 5 to 15 times the weight of the herbal ingredients, C1 to C4. Lower alcohols or mixed solvents thereof, preferably with water, hot water extraction, cold immersion at 0 to 120 ° C., preferably at 20 to 100 ° C. for about 1 hour to 1 day, preferably 2 hours to 5 hours Extraction method such as extraction, reflux cooling extraction, ultrasonic extraction or supercritical extraction, preferably extracted by hot water extraction method, the extract is filtered and then cooled and the filtrate can be lyophilized to obtain the mixed herbal extract of the present invention .

The present invention provides a pharmaceutical composition for the prevention and treatment of osteoporosis diseases containing the mixed herbal extract obtained by the above method as an active ingredient.

The composition for the prevention and treatment of osteoporosis diseases of the present invention, the extract is contained in an amount of 0.1 to 50% by weight based on the total weight of the composition.

However, the composition as described above is not necessarily limited thereto, and may vary according to the condition of the patient and the type and extent of the disease.

The composition comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Compositions comprising extracts according to the invention are formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Carriers, excipients and diluents which may be used in combination with the extract, and which may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin , Calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. Administration may be administered once a day or may be divided several times. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

In addition, it provides a health functional food for the prevention and improvement of osteoporosis disease containing the mixed herbal extract of the present invention as an active ingredient.

      The composition comprising the extract of the present invention can be used in various ways, such as drugs, food and beverages for the prevention and improvement of osteoporosis diseases. Foods to which the extract of the present invention may be added include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

Since the extract itself of the present invention has little toxicity and side effects, it is a drug that can be used safely even when taken for long periods of time.

       The extract of the present invention may be added to food or beverages for the purpose of preventing and improving osteoporosis disease. At this time, the amount of the extract in the food or beverage is generally added to the health food composition of the present invention to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 10 g based on 100 ml, preferably Can be added in a ratio of 0.3 to 1 g.

In addition to containing the extract as an essential ingredient in the indicated proportions, the health beverage composition of the present invention has no particular limitation on the liquid component, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like. Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

      In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

 The mixed herbal extracts of safflower, succinate and sine of the present invention not only increase the survival rate of MG-63 cells, osteoblasts, but also decrease the amount of bone (vaginal), estradiol and osteocalcin, which cause osteoporosis. Since it shows an increasing effect, it can be usefully used in the pharmaceutical composition and health functional food for the prevention and treatment of osteoporosis disease.

1 is a diagram showing the effect of increasing the survival rate of MG-63 cells by CRA,

Figure 2 is a diagram showing the effect of increasing the spinal bone mineral density (spinal BMD) reduced in osteoporosis-induced animals by CRA,

3 is a diagram showing the effect of increasing the bone mineral density (femoral BMD) of the femur reduced in osteoporosis-induced animals by CRA,

4 is a diagram showing the effect of increasing the bone mineral density (Tibial and Fibular BMD) of the tibia and fibula decreased in osteoporosis-induced animals by CRA,

5 is a diagram showing the effect of increasing the concentration of estradiol in the serum reduced in osteoporosis-induced animals by CRA,

Figure 6 is a diagram showing the effect of increasing the concentration of osteocalcin in the serum reduced in osteoporosis-induced animals by CRA.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, the contents of the present invention is not limited by the following Examples, Reference Examples and Experimental Examples.

Example 1 Preparation of Mixed Extract of Safflower, Sukji Sulfur and Sine

Wash 50g of safflower, 25g of Sukjihwang and 25g of sine which were purchased from Kyungdong Market ( http://www.kyungdongmart.com ) and wash them cleanly and place them in an electric brewing machine (Daewoong brewing machine, DWP5000M) and add 1.3ℓ of distilled water. Extracted. The extract was filtered through a filter paper (Advantec, 71101903), and the filtrate was concentrated under reduced pressure with a rotary vacuum distillation machine (EYELA, A-1000s) and lyophilized to obtain 15 g (yield: 15%) of a mixed extract of safflower, sperm sulphide and sine. . (Hereinafter referred to as "CRA")

Reference Example 1. Preparation of Experimental Materials

Fetal bovine serum (FBS) was purchased from Lonza co., Ltd., and Streptomycin, penicillin and DMEM medium were from Gibco co., Ltd. It was purchased and used.

Reference Example 2. Preparation of Laboratory Animals

The experimental animals were purchased from Hyochang Science, and 6-week-old Sprague-Dawley male rats (200-250 g) were used, and the feed and water were freely consumed. 2 ℃, relative humidity was maintained at 53 ± 3%, the contrast was turned on and off every 12 hours.

Reference Example 1. Culture of MG-63 Cells

MG-63 cells, which have the characteristics of human osteoblast-like cells purchased from the Korean Cell Line Research Foundation, were 10% FBS (heat inactivated fetal bovine serum) and 100 ug / ml strepto Cultured in an incubator with 5% CO _2, 100% humidity, 37 ° C using DMEM medium added with mycin (streptomycin) and 100 U / ㎖ penicillin (MG-63) The cells used cells in exponential growth phase.

Reference Example 2 Induction of Osteoporosis in Rats by Ovariectomy

2-1. anesthesia

General anesthesia was performed by intraperitoneal injection of 50 mg pentobarbital sodium (Entobal, Hallym Pharm.) Per kg.

2-1. Ovariectomy

The ovarian removal group uses 10% povidone iodine according to the usual preoperative procedures to clean and separate the surgical site and the surrounding area, and then cut the skin, muscles, and peritoneum of the lower abdomen to expose both ovaries, and then remove the ovaries and remove the resected site. After ligation with No. 4 silk, each layer of peritoneum, muscle, and skin was sealed with No. 3 silk (hereinafter referred to as 'OR').

On the other hand, the false ovarian removal group exposed both ovaries in the same manner as above, and returned to the abdominal cavity without resection of the ovaries, and then sutured each layer of the peritoneum, muscle, and skin with No. 3 silk (hereinafter referred to as SHAM). Naming).

The ovarian removal group and the false ovary removal group were used as experimental animals of Experimental Examples 2 to 3 after 3 weeks of surgery.

Reference Example 3. Statistical processing

Statistical significance was tested by one-way & two way ANOVA and post-hoc LSD of SPSS program (SPSS Inc., version 11.01).

Experimental Example 1 MG-63 Cells Survival rate measurement

In order to determine the effect of CRA prepared by the method of Example 1 on the survival rate of MG-63 cells, MTT assay ([3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide]) (Oh K. et al ., Effect of Rehmaniae glutinosa Libosh extracts on bone metabolism, Clinica Chimaica Acta, 334, 185-195, 2003).

MTT assay is based on mitochondrial dehydrogenase activity in living cells. The water-soluble tetrazolium salt MTT (yellow) is reduced to formazan crystals (purple). (formazan crystal) production is increased, the absorbance is measured high.

MG-63 cells cultured by the method of Reference Example 1 were placed in a 12-well plate, and the concentration of CRA dissolved in a control group of phosphate buffered saline (PBS) or phosphate buffered saline (PBS) (0, 0.01 mg / ml, 0.03 mg / Ml, 0.1 mg / ml) and incubated for 48 hours. 50 μl of MTT (2 mg / ml, Promega, 223613) was added to the culture solution and reacted for 2 hours. The reaction solution was centrifuged at 1000 rpm for 5 minutes to remove the supernatant. Then, 100 μl of DMSO (Dimethylsulfoxide, Sigma, C1619) was added to dissolve the formazan crystal in a plate shaker for 5 minutes, and then absorbed at 540 nm using a microplate reader (TECAN, I112904). The survival rate of MG-63 cells by CRA was calculated based on the average absorbance value of the control group as 100.

As a result, as shown in Figure 1, the CRA treatment group increased the survival rate of MG-63 cells at all treatment concentrations than the control group, especially CRA treatment group significantly at the concentration of 0.03mg / ㎖ and 0.1mg / ㎖ It was an increase. Through this, it was confirmed that CRA increases the survival rate of MG-63 cells (see Fig. 1).

Experimental Example 2 Measurement of Bone Mineral Density in Osteoporosis-induced Rats

In order to determine the effect of the CRA prepared by the method of Example 1 on the bone mineral density of the osteoporosis-induced rats by the method of Reference Example 2, the experiment was performed as follows using the method described in the literature (Dontas I et al . Protective effect of plant extract from Onobrychis ebenoides on ovariectomy-induced bone loss in rats. Maturitas. 53, 234-242, 2006)

Three weeks after the operation in the method of Reference Example 2, 0.9% physiological saline (positive control) in SHAM (10 mice / group) and 0.9% physiological saline (negative control) or CRA in OR daily weight (Kg) 100 mg / dose was administered orally for 3 weeks, followed by anesthesia using pentobarbital sodium (Entobal, Hallym Pharm.), Followed by dual energy radiation absorption of spine, femur, tibia and fibula in each experimental animal group. Bone mineral density was measured using a dual energy X-ray absorptiometry (PIXImus, Lunar).

As a result, as shown in Figure 2 to 4, in the CRA group, the bone density of the spine, femur, tibia and fibula increased compared to the negative control group, especially in the CRA group, the bone density of the femur, tibia and fibula was significantly higher than that of the negative control group. Showed an increase. Through this, it can be seen that CRA increases the amount of reduced bone mass, which is the cause of osteoporosis (see Figs. 2 to 4).

Experimental Example  3. Measurement of Hormone and Protein Content in Serum of Osteoporosis-Induced Rats

In order to determine the effect of the CRA prepared by the method of Example 1 on the serum content of the serum of the osteoporosis-induced rats by the method of Reference Example 2, the experiment was carried out as follows using the method described in the literature ((Suzuki N. et al . , A possible role of estrone produced in adipose tissues in modulating postmenopausal bone density, Maturitas, 22, 9-12, 1995; Srivastava AK, et al ., Development and application of a serum C-telopeptide and osteocalcin assay to measure bone turnover in an ovariectomized rat model, Calcif Tissue Int. 66 (6), 435-442, 2000).

Three weeks after the operation in the method of Reference Example 2, 0.9% physiological saline (positive control) in SHAM (10 mice / group) and 0.9% physiological saline (negative control) or CRA in OR daily weight (Kg) 100mg of sugar was orally administered for 3 weeks, and then anesthetized using pentobarbital sodium (Entobal, Hallym Pharmaceutical), and blood was collected from the subclavian vein of each experimental animal group before the procedure of Experimental Example 2 above. The blood was centrifuged at 3,000 rpm for 15 minutes to separate serum. Estradiol content in serum was used with Cota-A-Count estradiol RIA kit (DPC, TKE 21). Osteocalcin was used as OSTEOCALCIN MYRIA RIA kit (Techno genetics, 16099436).

As a result, as shown in Fig. 5 and 6, the CRA administration group was found to increase significantly the content of estradiol and osteocalcin in the serum compared to the negative control group. Through this, it was found that CRA increases the reduced estradiol and osteocalcin, which causes the osteoporosis (see FIGS. 5 to 6).

Through the above results, the CRA of the present invention increases the survival rate of osteoblasts and decreases the amount of bone density, which is one of the causes of osteoporosis, not only to suppress the loss of bone density, but also to reduce the cause of another osteoporosis. Increasing estradiol and osteocalcin was also found to inhibit bone resorption.

Examples of the formulation of the composition comprising the extract of the present invention will be described below, but the present invention is not intended to be limited thereto but merely to be described in detail.

Formulation Example 1 Preparation of Powder

300 mg CRA of Example 1

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

300 mg CRA of Example 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

300 mg CRA of Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

300 mg CRA of Example 1

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

300 mg CRA of Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve, the lemon flavor is added appropriately, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled into a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Food

CRA 1000 mg of Example 1

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

0.2 μg of vitamin B ₁₂

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 7 Preparation of Healthy Drink

CRA 300 mg of Example 1

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

0.25 g of vitamin B ₁

0.3 g of vitamin B ₂

Water quantification

After mixing the above components in accordance with a conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

Claims (6)

A pharmaceutical composition for the prevention and treatment of osteoporosis diseases containing a mixed herbal extract of safflower (Carthami Semen), Sukjiwang (Rehmanninae Rhizoma Preparat) and sinus (Amomi Fructus) as an active ingredient. The pharmaceutical composition of claim 1, wherein the extract is an extract available in a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The pharmaceutical composition according to claim 1, wherein the weight ratio of the mixed herbal medicine includes a weight ratio (w / w) of safflower (1-10): sucrose sulfur (1-5): sinus (1-5). , The method of claim 1, wherein the osteoporosis disease is osteoporosis type 1 (postmenopausal osteoporosis), type 2 osteoporosis (senile osteoporosis), idiopathic osteoporosis, endocrine and osteoporosis, digestive and osteoporosis, malignant and osteoporosis, drinking and drinking Pharmaceutical composition, characterized in that it comprises one or more diseases selected from the group consisting of osteoporosis, smoking and osteoporosis and drugs and osteoporosis.      A health functional food for the prevention and improvement of osteoporosis diseases containing a mixed herbal extract of safflower (Carthami Semen), Rehmanninae Rhizoma Preparat and Amomi Fructus as an active ingredient. A dietary supplement according to claim 5 which is a powder, granule, tablet, capsule or beverage.