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    • 专利摘要:
    PURPOSE: A herbal extract obtained by extracting Persicae Semen in alcohol or a mixed solvent of alcohol and water is provided. Therefore, the extract can be effectively used in prevention and treatment of various diseases related with growth hormone because it has high growth hormone releasing activity. CONSTITUTION: Alcohol or a mixed solvent of alcohol and water is added to Persicae Semen, hydrolyzed with a 0.1 to 2N acid or base solution, neutralized and then extracted with alcohol or a mixed solvent of alcohol and water. The alcohol or a mixed solvent of alcohol and water is selected from the group consisting of 10 to 100% ethyl alcohol, 10 to 100% methyl alcohol and spirit. The Persicae Semen has an LD50 value of 222.5g±7.5g/kg as measured by assays against mice.
    公开号:KR20030004042A
    申请号:KR1020020035770
    申请日:2002-06-25
    公开日:2003-01-14
    发明作者:김정숙；하혜경；정대영；이제현；김진숙
    申请人:한국 한의학 연구원；
    IPC主号:
    专利说明:

    Extract of herb for promoting release of growth hormone}
    [2] The present invention relates to a method for preparing Persicae Semen extract, to extracts obtained by the method and to pharmaceutical compositions and health foods comprising the same, more specifically to the addition of alcohol or a mixed solvent of alcohol and water To hydrolyzed and neutralized with an acid or a base solution, and to extract the extract of the phosphorus extracted by the addition of alcohol or a mixed solvent of alcohol and water, to the extract and the pharmaceutical composition and health food comprising the same as an active ingredient It is about.
    [3] Growth hormone (GH) is a protein secreted by the pituitary gland of animals and is a peptide hormone consisting of 191 amino acids. These growth hormones have been isolated from animals for a long time and have been used mainly for the treatment of dwarfism patients. They have various effects because they inhibit protein loss and delay physiological aging in older adults or the elderly. It can be used to treat diseases (Horber, FF and Haymond, MW, J. Clin. Invest ., 1990, 86, 265-272).
    [4] Continuous administration of growth hormone increases body weight, muscle, and fat weight to prevent aging-related diseases. Even when administered temporarily, plasma insulin and IGF-1 (Insulin-like growth factor-1) It is known to increase the concentration and decrease the protein concentration in urine (Kohrt, WM et al., Med. Sci. Sports Exerc ., 1992, 24, 832-837; Zachweija, JJ et al., Am. J. Physiol , 1994, 266 (Endocrinol. Metab. 29), E840-844), as well as increasing the weight and size of the liver and improving the synthesis of proteins in terminal tissues such as muscle, bone and connective tissue (Kaiser, FE). et al., J. Am. Geriatr. Soc ., 1991, 39, 235-240; Marcus, R. et al., J. Clin. Endocrino. Metab ., 1990, 70, 519-527).
    [5] Until now, the effects of simultaneous growth hormone releasing hormone (GHRH) and arginine on children and adults with congenital hypopituitarism have been shown to be effective. Growth hormone response was higher in children than in adults, and in patients with growth hormone deficiency than in many patients with pituitary hormone deficiency, and in patients with residual vascular components of the pituitary gland Higher results were obtained. Responsiveness of growth hormone to GHRH and arginine tends to decrease with age ( J. Clin. Endocrinol. Metab ., 2001, 86 (4), 1574-79), and endogenous growth hormone secretagogues. Ghrelin, a ligand, was effective when administered to growth hormone deficient rats ( Diabetes , 2001, 50 (2), 227-32). The administration of alrondron (alendronate) increases bone mineral density in postmenopausal women with postmenopausal osteoporosis because it maintains a relatively higher rate of formation than when arendronate is administered alone. The effect of MK-677 and arendronate on IGF-1 levels was increased in both MK-677 alone or in combination with arendronate and increased IGF-1 levels and alleviated decreased bone formation. The bone mineral density of the neck increased while the bone mineral density of the lumbar spine and hip did not increase. The metabolic effect of growth hormone through MK-677 reduced the direct inhibitory effect of arendronate on bone formation ( J. Clin. Endocrinol. Metab ., 2001, 86 (3), 1116-25).
    [6] In the past, growth hormone was mainly extracted from the cadaveric pituitary gland, but in the process it was found to cause the disease Creutzfeldt-Jakob disease in patients receiving growth hormone. As such, there is a risk that other diseases may be introduced into the patient during the separation process, and growth hormone isolated from the carcass has been limited in its use. With the development of genetic engineering, recombinant growth hormone produced using microorganisms can be produced and supplied in large quantities, and the risk of infection of other diseases is low, so the protein preparation including recombinant growth hormone occupies most of the market for dwarfism. The method of administering the recombinant recombinant growth hormone into the body is made by intramuscular injection using a syringe because the growth hormone itself is a huge protein and cannot be orally administered. As such, although recombinant growth hormone can be mass produced, attempts to develop oral preparations have been actively conducted in recent years due to the high cost required for a certain treatment period and the inability to use injection because it is impossible to orally administer. It is becoming.
    [7] The basic requirement for growth hormone preparations for oral administration is to have high absorption rate in the body and to have biological activity that can selectively stimulate growth hormone secretion. In order to have a high absorption rate in the body by oral administration, the smaller the molecular weight is advantageous, and to perform the physiological activity in the body smoothly, it must have a special physical and chemical properties.
    [8] Representatives that have been studied as substances with these basic requirements and have been developed up to the clinical trial stage include organically synthesized L-692, 429 ( Horm. Res. , 1993, 40, 109-115) and L-163, 191 ( PNAS). , 1995, 92, 7001-7005), and the peptide compounds GHRP-2 ( Endocrinology , 1994, 140, R9-R13), GHRP-1 ( J. Neuroendocrinology , 1994, 6, 185) and hexarelin (Hexarelin: His -D-2-methyl-Trp-Ala-Trp-D-Phe-Lys-NH2). The materials are newly researched and developed using the basic structure of GHRP-6 (His-D-Trp-Ala-Trp-D-Phe-Lys-NH2), a peptide compound commonly known in the early 1980s.
    [9] Recently, the approach of developing a new material has been in the spotlight by using a chemical library to search for a large amount of candidates, and in addition to the peptide library composed of short peptides and the natural world, Natural libraries consisting of many existing materials are also used to develop these new materials. Developing new drugs using such a wide variety of libraries is considerably more efficient than traditional approaches in terms of the time and cost of development, so developed pharmaceutical companies have long been active in developing new drugs using their libraries. come. Specifically, L-163, 191, and hexarelin may be referred to as substances developed by this approach.
    [10] Originally it varies from animal to animal, but in humans there is 44 amino acids of GHRH. The regulation of growth hormone secretion in the human body is a hormone called GHRH which promotes the secretion of growth hormone and somatostatin which inhibits the growth hormone secretion. It has been known to act to secrete
    [11] GHRP-6 is a useful synthetic peptide that is less effective than GHRH in biological activity but is highly compressed in terms of molecular weight size ( Journal of Endocrinology , 1995, 10 (1), 1-9). Since the development of GHRP-6, many researchers have reported that GHRP-6 does not act on the same receptors as GHRH, and the possibility of the presence of new receptors via these synthetic growth hormone secretagogues. Has been raised ( Endocrinology , 1989, 124, 2791). This possibility is demonstrated by the discovery of the cDNA sequence of growth hormone secretagogue receptors present in the pituitary archromediate and hypothalamus of pigs and humans by Merck researchers. ( Science , 1996, 273, 974-977). This fact proved the synergism between the previously found growth hormone secretagogue and the synthetic compound growth hormone secretagogue, and is more interested in the identification of substances that exist in the body and are expected to play the same role as GHRH. This situation is increasing.
    [12] L-692, 429 ( Science , 1993, 260, 1640-1643), developed by Merck, USA, has been shown to selectively secrete growth hormone, and has been shown to have an effective body absorption rate even in animal testing. However, the results of human experiments, unlike that of animals had a problem that the absorption is not very high when administered orally. Therefore, L-163, 191, which improved these problems, was developed. This is a compound that increased oral absorption rate, which was the biggest problem of the growth hormone secretagogues, which has been developed previously. Clinical trials are underway ( PNAS , 1995, 92, 7001-7005; Endocrinology , 1996, 137, 5284-5289). The results of these studies suggest that substances that stimulate the pituitary gland to secrete growth hormone may be useful clinically as growth hormone replacements, while developing new growth hormone secretagogues is not easy. Shows.
    [13] As part of such research, researches on finding substances that promote growth hormone secretion using natural extracts that are harmless to the human body are being conducted. Examples include alcoholic extracts of Astragalus root and 7-hydroxy-4'-methoxy-isoflavone, formononetin and stigmas-4-en-6β-ol-3-one (stigmast-4-en-6β-ol-3-one) and the like have been confirmed by the present inventors to stimulate growth hormone secretion (Korean Patent Nos. 257840 and 251519; Kim, JS and Kim, CS, Kor. J. Pharmacogn ., 1997, 28 (2), 75-79; Kim, JS et al., Kor. J. Pharmacogn. , 1996, 27 (4), 336-341), Regarding the growth hormone secretagogue composition containing one or more extracts selected from the group consisting of extracts from the ground, extracts from the dead, extracts from lilies, and gilyeong extracts (Korean Patent Application No. 1998-0053921), growth hormone secretion stimulating factor from natural herbal medicines Method and extract of water-soluble fast extract (Korean Patent Application No. 1998-014589) and glycoalkaloids A buffer saponin H1 growth hormone secretion stimulators of (asperosaponin H1) has been reported studies on (Republic of Korea Patent Application No. 1998-014590 No.).
    [14] Peach Kernel (Persicae Semen) is the seed of Prunuspersica Batsch or Prunus davidiana Maximowicz ( Rosaceae ). Seeds are used as herbal medicine and are called Persicae Semen.
    [15] Dried phosphorus contains at least 0.5% amigdalin (C 20 H 27 NO 11 ; 457.44) when quantified. The components of the phosphorus include amigdaline, emulsin, fatty oil, and the like. In addition, the chemical composition of the phosphorus is Prunin, (+)-Catechin ((+)-Catechin), Choline, Guanidine, Methylguanidine, (3R) -12'-Apo- β-carotene-3,12′-diol ((3R) -12′-Apo-β-carotene-3,12′-diol), (3S, 5R, 6S, 15Z) -5,6-epoxy-5, 6-dihydro-12'-apo-β-carotene-3 ((3S, 5R, 6S, 15Z) -5,6-Epoxy-5,6-dihydro-12'-apo-β-carotene-3) , 12'-diol, (3S, 5R, 6S, 9Z, 13Z ')-5,6-epoxy-5,6-dihydro-12'-apo-β-carotene-3,12'-diol (12 '-diol, (3S, 5R, 6S, 9Z, 13Z')-5,6-Epoxy-5,6-dihydro-12'-apo-β-carotene-3,12'-diol), (3S, 5R , 8R, all-E) -5,8-epoxy-5,8-dihydro2'-apo-β-carotene-3,12'-diol ((3S, 5R, 8R, all-E) -5, 8-Epoxy-5,8-dihydro12'-apo-β-carotene-3,12'-diol), (3S, 5R, 8S, all-E) -5,8-epoxy-5,8-dihydro12 '-Apo-β-carotene-3,12'-diol ((3S, 5R, 8S, all-E) -5,8-Epoxy-5,8-dihydro12'-apo-β-carotene-3,12' -diol), 9-cis-β-Carotene, Cryptoxanthin, Persicaxanthin, Zeaxanthin anthin (all E-form)), α-Carotene, β-Carotene (β-Carotene (all-trans)), Gibberellin A32, Gibberellin A5 (Gibberellin A5), Xanthanol, (-)-Epicatechin-3-O-gallate, (-)-Epicatechin-3-O-gallate, Afzelin, Astragalin, Hesperetin5- Hesperetin-5-O-β-D-glucopyranoside, Hyperside, Isoquercitrin, Camperol-3-O-β-D-Lu Kaempferol-3-O-β-D-rutinoside, Multiflorin A, Multiflorin B, Multinoside A, Quercetin-3-O-β-D -Quercetin-3-O-β-D-diglucopyranoside, Quercetin-3-O-galactopyranosyl-7-O-Diglucopyranoside (Quercetin-3-O-galactopyronosyl-7 -O-diglucopyranoside), quercetin-3-O-glucopyranosyl-7-O-diglucopyranoside (Quercetin-3-O-glucopyranosyl -7-O-diglucopyranoside), quercitrin, rutin ( Rutin), Trifolin, Mevalonic acid, Benzaldehyde, Caffeic acid, Chlorogenic acid, dl-Mandelic acid, Arginine , Creatine, Guanidinoacetic acid, γ-Guanidinobutyramide, γ-Guanidinobutyric acid, γ-Guanidino-Gropionic Acid (γ-guanidinopropionic acid), isoxanthanol, tomentosin, xantatin, xanthatin, xanthinin, xanthumin, campesterol, daucosterol ), Stigmasterol, δ-5-avenasterol and the like.
    [16] Doin's medicinal effect is combined with anti-inflammatory oral colloidal drugs in menstrual pain, dysmenorrhea, and bruises in Chinese medicine. In the private sector, dried constipated peach buds are used to boil, and sweat leaves are put in bath water. Doin has little smell and tastes bitter and oily.
    [17] Prescriptions for the use of Toin are Gyejiointang, Gumi-Shinhwan, Doinseunggi-tang, Dointang, and Dokbyeongtang. And Tong Kyung-hwan. Applicable conditions include stool secretion, shooter, postpartum back pain, postpartum swelling, upper limit, upper limit congestion, small abdominal pain, childhood ecchymosis, ten-degree sieve, premenorrhea, five o It is used for symptoms such as bleeding, drowsiness pain, gongga, blood value, and high blood pressure. Doin's numerical methods include peeling, Doinni, Dointang, Sutang, Chodoin, and Tangpo.
    [18] The effects of doin from recent scientific research have shown that they have a protective effect on UV-induced cytotoxicity of normal human keratinocytes and fibroblasts ( Archives of Phamacial Research , 2000, 23 (4), 396-400) reported to contain inhibitors for mutagens (Yakugaku Zasshi-J, Pharmaceutical Society of Japan, 1992, 112 (12), 934-9), hypocholesterolemic mice Have been reported to lower cholesterol levels ( Kor. J. Pharmacogn ., 1990, 21 (2), 153-157) and scavenging oxygen radicals (Yakugaku Zasshi-J. Pharmaceutical Society of Japan , 1985, 105 (9), 895-901).
    [19] To this end, known to have a variety of efficacy has been prepared in the form of a mixture with other herbal medicines to date. There is a metastasis inhibiting composition (Registration No. 291630), a composition for treating gynecological diseases and female obesity (Registration No. 232671), and a method for separating and purifying isoflavone (US 5679806), cherry Bioflavonoids are applied to COX inhibitors and anti-inflammatory agents (US 194469), allergy inhibitors (US 6180106), anti- analgesic anti-inflammatory agents (US 5908628), and the like. It also increases blood pressure circulation (US 5942233), and is used as an antioxidant (WO 039249A1) in addition to dietary supplements (WO 103714A1). Other applications include hair regrowth, phosphatase inhibitors (JP 103293A2), and health teas (JP 8073369A2). It is a compounding agent that is applied to rheumatism, burns and wound healing.
    [20] Accordingly, the present inventors have completed the present invention by studying the physiological activity of the extract of the cabbage, revealing that the cabbage extract acts as a growth hormone secretagogue to increase the production of growth hormone.
    [21] It is an object of the present invention to provide a method for preparing a seed extract having growth hormone secretagogue activity, a seed extract obtained by the method, and a pharmaceutical composition and health food containing the same as an active ingredient.
    [1] 1 is a graph showing the growth hormone was measured at regular intervals from the tail vein after administering the extract of the present invention to the jugular vein of the rat.
    [22] In order to achieve the above object, the present invention provides a method for producing a phosphorus extract.
    [23] In addition, the present invention provides a phosphorus extract having growth hormone secretagogue activity prepared by the above method.
    [24] The present invention also provides a pharmaceutical composition for promoting growth hormone secretion containing the extract as an active ingredient.
    [25] In addition, the present invention provides a growth hormone secretion promoting health food containing the extract as an active ingredient.
    [26] Hereinafter, the present invention will be described in detail.
    [27] The present invention provides a method for producing a phosphorus extract.
    [28] Specifically, the present invention provides a method of preparing the phosphorus extract by adding alcohol or a mixed solvent of alcohol and water and immersing the phosphorus at 5 to 40 ° C. The aqueous alcohol solution may be selected from the group consisting of 10 to 100% ethyl alcohol (ethyl alcohol), 10 to 100% methyl alcohol (methyl alcohol) and spirits.
    [29] In another aspect, the present invention provides a method for preparing a causal extract by hydrolyzing and neutralizing the phosphorus with an acid or a base, and then adding an alcohol or a mixed solvent of alcohol and water. The acid or base is a concentration of 0.1 to 2 N Preferably, the acid may be selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid and the base may be selected from the group consisting of sodium hydroxide and potassium hydroxide. According to this method, the bound compound of the plant, which has been bound to various substituents such as sugar or other alkyl groups, is hydrolyzed, and the phosphorus extract is included as a compound of the free form of the plant. The aqueous alcohol solution may be selected from the group consisting of 10 to 100% ethyl alcohol (ethyl alcohol), 10 to 100% methyl alcohol (methyl alcohol) and spirits.
    [30] In addition, the present invention provides a phosphorus extract having growth hormone secretagogue activity prepared by the above method.
    [31] The present inventors confirmed whether the extract of the present invention has growth hormone secretagogue activity. First, pituitary cells were removed from Sprague-Dawley rats, 4-5 weeks of age, under aseptic manipulation, gently washed and crushed to separate pituitary cells. The pituitary extract of the present invention was added to the isolated pituitary cells, and in the comparison group, rat rat growth hormone secretion factor (rat GRF, Bachem Co., Torrance, CA, USA), which is a reference substance for growth hormone secretion, was added. Incubated at ℃. After incubation, the supernatant was separated by centrifugation, and the amount of growth hormone was measured using a rat growth hormone RIA kit.
    [32] As a result, the extract of the present invention acts on the pituitary cells to promote the secretion of growth hormone, and it was confirmed that the growth hormone has a superior secretory promoting effect than the rat growth hormone secretion factor used as a comparison group (see Table 1 ). .
    [33] In addition, growth hormone secretion experiments using animal models, the extract was found to be a potent increase in growth hormone in vitro ( in vitro ) as well as in vivo (see Table 2 ).
    [34] The present invention also provides a pharmaceutical composition for promoting growth hormone secretion containing the extract as an active ingredient.
    [35] The pharmaceutical composition for promoting growth hormone secretion of the present invention contains the extract of Doin as an active ingredient. The causal extract can be administered orally or parenterally during clinical administration and can be used in the form of a general pharmaceutical formulation. In other words, the extract of the present invention can be administered in a variety of oral and parenteral formulations in actual clinical administration, when formulated, diluents such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants commonly used, or Formulated using excipients. Solid formulations for oral administration include tablets, pills, powders, granules, and capsules, and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose and gelatin, etc. Mix is prepared. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral administration include suspensions, 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. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. As the non-aqueous solvent and the suspension solvent, 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, glycerol and gelatin may be used.
    [36] Dosage units may contain, for example, one, two, three or four times the individual dosage, or they may contain 1/2, 1/3 or 1/4 times. The individual dosage preferably contains an amount in which the effective drug is administered at one time, which usually corresponds to all, 1/2, 1/3 or 1/4 times the daily dose.
    [37] The effective dose of the cabbage extract is 0.1 to 40 mg / kg, preferably 0.4 to 4 mg / kg, and may be administered 1-6 times a day.
    [38] Toxicity experiments of oral, intraperitoneal, and subcutaneous injections of the doin extract of the present invention were performed in mice, and 50% lethal dose (LD 50 ) by intraperitoneal and subcutaneous injection toxicity tests was at least water extract 222.5 ± It turned out to be a safe material that is greater than 7.5 g / kg.
    [39] In addition, the present invention provides a growth hormone secretion promoting healthy food containing the extract as an active ingredient.
    [40] In the present specification, the health food is a food that improves the function of the general food by adding a phosphorus extract to the general food, and the phosphorus extract may be added to the general food, or may be prepared by encapsulation, powdering, and suspension. When ingested, it brings a specific effect on health, and unlike the general medicine because the food is a raw material has the advantage that there is no side effect that can occur during long-term use of the drug.
    [41] When the extract of the present invention is used as a food additive, the extract of the phosphorus may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient can be suitably determined according to the purpose of use (prevention, health or therapeutic treatment). Generally, the causal extract may be added in an amount of 0.0001 to 10% by weight, preferably 0.1 to 5% by weight based on the raw materials in the manufacture of the food or beverage. However, in the case of prolonged ingestion for health and hygiene purposes or for health control purposes, the amount may be below the above range. In addition, the health food of the present invention contains an extract of isotope within the toxic range measured upon use in the pharmaceutical composition.
    [42] There is no particular limitation on the kind of food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes. Specifically, the health food containing the extract of the phosphorus is a health food and favorite products such as juice, tea, jelly, juice made with the main component of the phosphorus, and folk remedies for the purpose of dysmenorrhea, bruise pain, etc. Can be. In addition, it can be used in various oriental medicine prescriptions such as Gyejidointang, Doinseunggitang, Dointang, and Tongkyeonghwan.
    [43] Hereinafter, the present invention will be described in detail by way of examples.
    [44] However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.
    [45] Example 1 Preparation of Doin Extract
    [46] 50 g of the dried fruit powder was added to 500 ml of an aqueous 70% methanol solution and stirred at room temperature for 2 days. The solution obtained through the stirring process was filtered through a filter paper, concentrated under reduced pressure to remove methanol, and then lyophilized to remove moisture to prepare a extract of the present invention.
    [47] Example 2 Preparation of Doin Extract 2
    [48] 0.1 g of phosphorus dried powder was added to 1 ml of 1 N HCl aqueous solution and reacted at 100 ° C. for 2 hours. After hydrolysis for 2 hours and neutralized, the reaction solution was lyophilized to remove moisture, and then, the same extract as in Example 1 was prepared.
    [49] Preparation Example 1 Preparation of Injection Solution
    [50] Injection solution containing 10 mg of the active ingredient was prepared by the following method.
    [51] 100 g of 1 g of phosphorus extract, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.
    [52] The components of the injection solution are as follows.
    [53] Ginseng extract 1 g
    [54] 0.6 g sodium chloride
    [55] 0.1 g of ascorbic acid
    [56] Distilled Water Determination
    [57] Preparation Example 2 Preparation of Syrup
    [58] Syrup containing Doin extract of the present invention as an active ingredient 2% (weight / volume) is prepared by the following method.
    [59] Cedar extract, saccharin and sugar were dissolved in 80 g of warm water. After the solution was cooled, a solution consisting of glycerin, spices, ethanol, sorbic acid and distilled water was prepared and mixed therein. Water was added to this mixture to 100 ml.
    [60] The components of the syrup are as follows.
    [61] Ginseng extract 2 g
    [62] Saccharin 0.8 g
    [63] 25.4 g per
    [64] Glycerin 8.0 g
    [65] 0.04 g of spices
    [66] Ethanol 4.0 g
    [67] 0.4 g of sorbic acid
    [68] Distilled Water Determination
    [69] Preparation Example 3 Manufacturing Method
    [70] A tablet containing 15 mg of active ingredient is prepared by the following method.
    [71] It was mixed with 250 g of cabbage extract, 175.9 g of lactose, 180 g of potato starch, and 32 g of colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into a tablet.
    [72] The components of the tablet are as follows.
    [73] 250 g of cabbage extract
    [74] Lactose 175.9 g
    [75] 180 g potato starch
    [76] 32 g of colloidal silicic acid
    [77] 10% gelatin solution
    [78] Potato Starch 160 g
    [79] 50 g of talc
    [80] 5 g of magnesium stearate
    [81] Preparation Example 4 Manufacturing Method of Food and Beverage
    [82] The present inventors prepared a food or beverage composition containing the extract as an active ingredient as follows.
    [83] <4-1> Preparation of Biscuits
    [84] Force Class 1 88 kg
    [85] Gravity First Class 76.4 ㎏
    [86] 16.5 kg per white
    [87] 2.5 kg of salt
    [88] 2.7 kg of glucose
    [89] Palm shortening 40.5 kg
    [90] 5.3 kg of ammo
    [91] Medium kg 0.6 kg
    [92] 0.55 kg sodium bisulfite
    [93] Rice flour 5.0 kg
    [94] Vitamin B1 0.003 kg
    [95] 0.003 kg of vitamin B2
    [96] Milk Flavor 0.16 ㎏
    [97] 71.1 kg of water
    [98] Whole milk powder 4 kg
    [99] Substitute powder 1 kg
    [100] 0.1 kg of calcium phosphate
    [101] Spray salt 1 kg
    [102] 25 kg of spray oil
    [103] 0.1-0.5 kg of cabbage extract
    [104] Biscuits were prepared using conventional methods with the above composition and content.
    [105] <4-2> Preparation of Beverage
    [106] 522 mg of honey
    [107] Chioctosanamide 5 mg
    [108] Nicotinamide 10 mg
    [109] Riboflavin Sodium Hydrochloride 3 mg
    [110] Pyridoxine hydrochloride 2 mg
    [111] Inositol 30 mg
    [112] Orthoic acid 50 mg
    [113] Peach Extract 0.48-1.28 mg
    [114] 200 ml of water
    [115] With the above composition and content, drinks were prepared using conventional methods.
    [116] Experimental Example 1 Growth Hormone Secretion Activity Test Using Doin Extract
    [117] <1-1> Pituitary Cell Culture
    [118] The pituitary gland was removed from Sprague-Dawley rats at 4-5 weeks of age under aseptic manipulation, followed by collagenase (Gibco co., Grand Island, NY) and hyaluronidase (Sigma co). , St. Louis, Mo.) rat rat pituitary cells were isolated. The isolated pituitary cells were suspended in Dulbecco's Modified Eagle's medium containing 10% horse serum, 2.5% fetal bovine serum (FBS), and antibiotics. Incubated at 37 ° C., 5% CO 2 ).
    [119] <1-2> Growth hormone secretion promoting activity experiment
    [120] The pituitary cells isolated in Experimental Example <1-1> were cultured and then suspended in a test solution. Medicinal herbs were added to pituitary cells at a concentration of 1 mg / ml, and rat growth hormone secretion factors (rat GRF, Bachem Co., Torrance, CA, USA), which are reference substances for growth hormone secretion, were 300 nM and 500 nM. , 1,000 nM was added and the test solution was added to a total volume of 1 ml and incubated at 37 ° C. for 15 minutes. After incubation, the supernatant was separated by centrifugation, and stored at -70 ° C to measure growth hormone.
    [121] Growth hormone measurement was performed using a rat growth hormone RIA kit (rat GH RIA kit, Amersham, Buckinghamshire, England). The growth hormone contained in a constant amount of 125 I-labeled growth hormone and the test substance stored in the above was tested by different reactions of the growth hormone-antibody complex (GH) by a competitive response to an anti-GH antibody. -Ab complex) was formed and precipitated by addition of a second antibody, and the radioactivity of 125 I was measured from the precipitate. The results are shown in Table 1 below.
    [122] Added concentrationGrowth hormone concentration (ng / ml) Control023.7 Comparative group * 300 nM38.2 500 nM57.7 1,000 nM146.6 Doin1 mg medicine / ml272.8
    [123] The rat growth hormone secretion factor was used as a comparison group.
    [124] As a result, as can be seen in Table 1 , the extract of the present invention acts on the pituitary cells to promote the secretion of growth hormone, and the growth hormone secretion effect is superior to the rat growth hormone secretion factor used as a comparison group It was confirmed.
    [125] Experimental Example 2 Growth Hormone Induction Rat
    [126] 8-week-old SD rats were anesthetized at a concentration of 50 mg / kg of pentobarbital, and then, the extract was dissolved in physiological saline at a concentration of 1 mg / kg of dry medicine, and administered to the jugular vein of the rat. It was. After bleeding from the tail jugular vein at regular intervals, the secreted growth hormone was measured using the RIA kit. Measurement of the concentration of growth hormone in the blood in the same manner as in Experimental Example 1 Rat growth hormone RIA kit (rat GH RIA kit, Amersham, Buckinghamshire, England) was performed. A certain amount125Growth hormones contained in the I-labeled growth hormone and the test substance stored above are different amounts of growth hormone-antibody complex (GH-Ab complex) by a competitive reaction to the anti-GH antibody. And precipitated by adding a second antibody to the precipitate125The radioactivity of I was measured and the corresponding concentration was calculated using a standard curve.
    [127] MinutesControl group (ng / ml)Doin (ng / ml) 03.6676.694 103.2994.314 202.8158.549 303.3659.110 452.5664.103 602.5933.209 903.27016.302 1202.2671.561
    [128] As shown in Table 2 above, the doin extract of the present invention was administered to the jugular vein of the rat at a concentration of 1 mg / kg of dry medicinal herbs using an animal, and the growth hormone of the separated plasma was measured by collecting blood at regular intervals. As a result, compared with the control group, the drug showed an increase of about 2.5 times after 30 minutes of drug administration, and an increase of about 5 times after 90 minutes, and the growth hormone secretion ability of the cabbage extract was measured within 2 hours ( FIG. 1 ).
    [129] Experimental Example 3 Oral Acute Toxicity in Rats
    [130] Acute toxicity test was performed using 6-week-old specific pathogen-free (SPF) SD rats. Two animals per group were suspended in a 0.5% methylcellulose solution in each of the extracts of the present invention and administered once orally at a dose of 1 g / kg / ml. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed. Necropsy was performed to observe abdominal and thoracic organ abnormalities.
    [131] As a result, no significant clinical symptoms or dead animals were noted in all animals treated with the test substance, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings. As a result, all of the extracts of the present invention do not show a toxic change up to 3 g extract / kg in mice, and the minimum lethal dose (LD 50 ) of oral administration was determined to be a safe substance with an extract of 222.5 ± 7.5 g / kg or more.
    [132] As described above, the extract of the present invention exhibits a very high growth hormone secretagogue activity, and the raw material of the extract is harmless to the human body, which has been used for a long time, and has good absorption rate, thereby treating various diseases related to growth hormone. And can be usefully used for prevention.
    权利要求:
    Claims (6)
    [1" claim-type="Currently amended] A process for producing a causal extract, which is extracted by adding alcohol or a mixed solvent of alcohol and water to Persicae Semen, or hydrolyzing and neutralizing with an acid or base solution, and then adding a mixed solvent of alcohol or alcohol and water.
    [2" claim-type="Currently amended] The method of claim 1, wherein the alcohol or a mixed solvent of alcohol and water is selected from the group consisting of 10 to 100% ethyl alcohol, 10 to 100% methyl alcohol and spirits Method of Preparation of Extracts.
    [3" claim-type="Currently amended] The method of claim 1, wherein the acid or base solution is a method for producing a sterilized extract, characterized in that the concentration of 0.1 to 2 N.
    [4" claim-type="Currently amended] Extract of doujinshi prepared by the method of any one of claims 1 to 3.
    [5" claim-type="Currently amended] A pharmaceutical composition for promoting growth hormone secretion comprising the extract of Doin in accordance with claim 4 as an active ingredient.
    [6" claim-type="Currently amended] Growth hormone secretion promoting health food containing the extract of claim 4 as an active ingredient.
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    同族专利:
    公开号 | 公开日
    KR100518686B1|2005-11-08|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    法律状态:
    2001-06-26|Priority to KR20010036534
    2001-06-26|Priority to KR1020010036534
    2002-06-25|Application filed by 한국 한의학 연구원
    2003-01-14|Publication of KR20030004042A
    2005-11-08|Application granted
    2005-11-08|Publication of KR100518686B1
    优先权:
    申请号 | 申请日 | 专利标题
    KR20010036534|2001-06-26|
    KR1020010036534|2001-06-26|
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