KR100497945B1 - Extract of herb for promoting release of growth hormone - Google Patents

Abstract

The present invention relates to a method for producing a tree extract (Akebiae Caulis), to a tree extract obtained by the method and to pharmaceutical compositions and health foods comprising the same, more specifically to the barrel to add alcohol or a mixed solvent of alcohol and water or To hydrolyze and neutralize with an acid or base solution, and to extract the neck extract extracted by the addition of alcohol or a mixed solvent of alcohol and water, the neck extract obtained by the method and a pharmaceutical composition and health food comprising the same as an active ingredient It is about. Throat extract of the present invention shows a strong growth hormone secretagogue activity and can be used as a growth hormone secretagogue drugs and foods because it is secured as a natural drug.

Description

Extract of herb for promoting release of growth hormone}

The present invention relates to a method for producing a tree extract (Akebiae Caulis), to a tree extract obtained by the method and to pharmaceutical compositions and health foods comprising the same, more specifically to the barrel to add alcohol or a mixed solvent of alcohol and water or To hydrolyze and neutralize with an acid or base solution, and to extract the neck extract extracted by the addition of alcohol or a mixed solvent of alcohol and water, the neck extract obtained by the method and a pharmaceutical composition and health food comprising the same as an active ingredient It is about.

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).

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).

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 growth hormone deficiency was higher in patients with growth hormone deficiency than in many types of pituitary hormone deficiency, and patients with residual vascular components of the pituitary gland Obtained higher results. 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).

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.

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.

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.

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.

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

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.

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, which is a compound that increased the oral absorption rate, which is the biggest problem of the growth hormone secretagogues that have been developed previously, and has been reported to have an absorption rate of more than 60% when orally administered to dogs. 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.

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.).

The keg (Akebiae Caulis) is a stem of the Akebia quinata Decaisne or other cognates (vines and Lardizabalaceae ) and has been cut off horizontally. The herbal medicine uses the same vegetation and stems of creeping vines, and is called a wooden barrel (Akebiae Caulis, Akebiae Lignum).

The constituents of the tree trunk are saponins (saponins), which contain a large number of akeboside saponins and pericarp saponins. In addition, the other chemicals contained in the barrel include Cyanidin-3-p-coumaroylglucopyranoside, β-Sitosterol, Orenolic acid, 3-α-Akebonoic acid, Akebonoic acid, Arjunoic acid, Quinatic acid, Mesembryanthemoidigenic acid, Headrazinine 3-O-β-D-Xylopyrronosyl- (1-3) -α-L-arabinopyronoside (Hedragenin-3-O-β-D-xylopyranosyl- (1-3) -α- L-arabinopyranoside), orenolic acid-3-O-β-D-glucopyranosyl (1-2) -α-L-arabinopyranoside (Oleanolic acid-3-O-β-D-glucopyranosyl (1 -2) -α-L-arabinopyranoside), noarjunolic acid-28-O-β-D-xylpyranosyl- (1-3) -α-L-α-L-rhamnopyranosyl- (1-4 ) -β-D-glucopyranosyl- (1-6) -β-D-glucopyranoside (norarjunolic acid-28-O-β-D-xylopyranosyl- (1-3) -α-L-α -L-rhamnopyranosyl- (1-4) -β-D-glucopyranosyl- (1-6) -β-D-glucopyranos ide), norarjunolic acid, quillaic acid, akebia saponin D, eleutheroside K, quinatoside A, quinatoside B (Quinatoside B), Quinatoside C, Quinatoside D, Pericarp saponin K, Pericarp saponin C, Pericarp saponin G G), Akebia saponin B, Akebia saponin A, Akebia saponin C, Akeboside Stk, Akeboside Stj Akeboside Sth, Hederasaponin B, Akebia saponin G, Akebia saponin F, Percarp saponin J2, Percarp saponin J2 Percarp saponin J3, Akebia saponin E, Stigmasterol, and the like.

Its medicinal effect is used as diuretic for edema, nephritis, urethritis and also for dysmenorrhea. The neck is almost odorless and tastes slightly awkward

The effects of neck pain from recent scientific studies have reported cytotoxic effects on human cancer cell lines ( Kor. J. Phamacogn ., 1999, 30 (2), 105-110).

As such, the barrel known to have various effects has been manufactured in the form of a mixture with other herbal medicines. Herbal medicine for preventing and treating liver cancer (Registration No. 239879), composition for treating gynecological diseases and women's obesity (Registration No. 232671), diabetes treatment and herbal medicine-derived dietary fiber (Registration No. 053436), etc. Developed. In addition, tripepene derivatives are synthesized and patented for use in nephritis (US 5885992), and are used as anti-inflammatory, anti-cancer, cosmetics, etc. as a combination, and as a therapeutic agent for chronic hyperosteogeny (CN 1133731A).

Thus, the present inventors have completed the present invention by studying the physiological activity of the extract of the neck, revealing that the extract acts as a growth hormone secretagogue to increase the production of growth hormone.

It is an object of the present invention to provide a method for preparing a throat extract having growth hormone secretagogue activity, a throat extract obtained by the method and a pharmaceutical composition and health food containing the same as an active ingredient.

In order to achieve the above object, the present invention provides a method for producing a barrel extract.

The present invention also provides a neck extract having growth hormone secretagogue activity prepared by the above method.

In addition, the present invention provides a pharmaceutical composition for promoting growth hormone secretion containing the neck extract as an active ingredient.

In addition, the present invention provides a growth hormone secretion promoting health food containing the throat extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a method for preparing a throat extract.

Specifically, the present invention provides a method for preparing the neck extract by adding an alcohol or a mixed solvent of alcohol and water and dipping 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.

In another aspect, the present invention provides a method for preparing a neck extract by hydrolyzing and neutralizing the neck with an acid or a base, and then adding an alcohol or a mixed solvent of alcohol and water. At this time, the acid or base is preferably in a concentration of 0.1 to 2 N, 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. have. 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 keg extract is included as a free form of the compound 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.

The present invention also provides a neck extract having growth hormone secretagogue activity prepared by the above method.

The present inventors confirmed whether the extract of the present invention has a 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. Throat extract of the present invention was added to the pituitary cells isolated from the above, and after the rat growth hormone secretion factor (rat GRF, Bachem Co., Torrance, CA, USA), which is a reference substance for growth hormone secretion, was added 37 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.

As a result, the throat extract of the present invention acted on the pituitary cells to promote the secretion of growth hormone, and it was confirmed that the growth hormone was much better in promoting the secretion than the rat growth hormone secretion factor used as a comparison group (see Table 1 ). .

In addition, growth hormone secretion experiments using animal models, neck extract was found to increase the growth hormone strongly in vitro ( in vitro ) as well as in vivo (see Table 2 ).

In addition, the present invention provides a pharmaceutical composition for promoting growth hormone secretion containing the neck extract as an active ingredient.

The growth hormone secretagogue pharmaceutical composition of the present invention contains the throat extract as an active ingredient. The neck extract can be administered orally or parenterally during clinical administration and can be used in the form of a general pharmaceutical formulation. That is, the neck extract of the present invention may be administered in various oral and parenteral dosage forms in actual clinical administration, and when formulated, diluents such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants that are commonly used, or Formulated using excipients. Solid preparations 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 in the neck extract. 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.

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.

The effective dose of the throat extract is 0.1-40 mg / kg, preferably 0.4-4 mg / kg, and can be administered 1-6 times a day.

Toxicity experiments of oral administration, intraperitoneal administration, and subcutaneous injection of the throat extract of the present invention to mice were conducted. As a result, 50% lethal dose (LD ₅₀ ) by intraperitoneal administration and subcutaneous injection toxicity test was at least 25.95 ±. It has been found to be a safe substance of more than 2.89 g / kg.

In addition, the present invention provides a growth hormone secretion-promoting health food containing a throat extract as an active ingredient.

In the present specification, the health food is a food that improves the function of the general food by adding a neck extract to the general food, and the neck 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.

When using the wood extract of the present invention as a food additive, the wood extract is added as it is or used with other foods or food ingredients, it can be suitably used according to a conventional method. The blending amount of the active ingredient can be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). In general, the keg extract may be added in an amount of 0.0001 to 10% by weight, preferably 0.1 to 5% by weight relative to the raw material 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 a throat extract within the toxic range measured upon use in the pharmaceutical composition.

There is no particular limitation on the kind of food. Examples of foods to which the neck extract can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinks, teas, drinks Alcoholic beverages and vitamin complexes. Specifically, the health food containing the extract of the neck is a health food and favorite products such as juice, tea, jelly, juice made mainly of the neck, and folk remedies for the purpose of edema, nephritis, urethritis, etc. Can be mentioned.

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

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example 1 Preparation of Bark Extract 1

50 g of the dry wood powder was added to 500 ml of 70% aqueous methanol solution and stirred for 2 days at room temperature. 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 neck extract of the present invention.

Example 2 Preparation of Bark Extract 2

0.1 g of dry wood powder was placed in 1 ml of 1 N HCl aqueous solution and reacted at 100 ° C for 2 hours. After neutralizing the hydrolyzate for 2 hours, the reaction solution was lyophilized to remove moisture, and then the extract of the present invention was prepared in the same manner as in Example 1.

Preparation Example 1 Preparation of Injection Solution

Injection solution containing 10 mg of the active ingredient was prepared by the following method.

1 g of a wooden barrel extract, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.

The components of the injection solution are as follows.

1 g of throat extract

0.6 g sodium chloride

0.1 g of ascorbic acid

Distilled Water Determination

Preparation Example 2 Preparation of Syrup

Syrup containing the neck extract of the present invention as an active ingredient 2% (weight / volume) is prepared by the following method.

Barrel 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.

The components of the syrup are as follows.

2 g of throat extract

Saccharin 0.8 g

25.4 g per

Glycerin 8.0 g

0.04 g of spices

Ethanol 4.0 g

0.4 g of sorbic acid

Distilled Water Determination

Preparation Example 3 Manufacturing Method

A tablet containing 15 mg of active ingredient is prepared by the following method.

It was mixed with 250 g of a tree 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.

The components of the tablet are as follows.

250 g throat extract

Lactose 175.9 g

180 g potato starch

32 g of colloidal silicic acid

10% gelatin solution

Potato Starch 160 g

50 g of talc

5 g of magnesium stearate

Preparation Example 4 Manufacturing Method of Food and Beverage

The present inventors prepared a food or beverage composition containing the throat extract as an active ingredient as follows.

<4-1> Preparation of Biscuits

Force Class 1 88 kg

Gravity First Class 76.4 ㎏

16.5 kg per white

2.5 kg of salt

2.7 kg of glucose

Palm shortening 40.5 kg

5.3 kg of ammo

Medium kg 0.6 kg

0.55 kg sodium bisulfite

Rice flour 5.0 kg

Vitamin B1 0.003 kg

0.003 kg of vitamin B2

Milk Flavor 0.16 ㎏

71.1 kg of water

Whole milk powder 4 kg

Substitute powder 1 kg

0.1 kg of calcium phosphate

Spray salt 1 kg

25 kg of spray oil

Neck barrel extract 0.1-0.5 kg

Biscuits were prepared using conventional methods with the above composition and content.

<4-2> Preparation of Beverage

522 mg of honey

Chioctosanamide 5 mg

Nicotinamide 10 mg

Riboflavin Sodium Hydrochloride 3 mg

Pyridoxine hydrochloride 2 mg

Inositol 30 mg

Orthoic acid 50 mg

Neck Bark Extract 0.48-1.28 mg

200 ml of water

With the above composition and content, drinks were prepared using conventional methods.

Experimental Example 1 Growth Hormone Secretion Activity Test

<1-1> Pituitary Cell Culture

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 ).

<1-2> Growth hormone secretion promoting activity experiment

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.

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.

Added concentration Growth hormone concentration (ng / ml) Control 0 23.7 Comparative group ^* 300 nM 38.2 500 nM 57.7 1,000 nM 146.6 Neck 1 mg medicine / ml 271.5

The rat growth hormone secretion factor was used as a comparison group.

As a result, as shown in Table 1 , the neck 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.

Experimental Example 2 Growth Hormone Induction Rat

Eight weeks-old Sprague-Dawley rats were anesthetized at a concentration of 50 mg / kg of pentobarbital, and the neck extract was dissolved in physiological saline at a concentration of 1 mg / kg of dry medicine and administered to the rat jugular vein. Physiological saline was administered. 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. 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 then the concentration was calculated using a standard curve by measuring the radioactivity of ¹²⁵ I from the precipitate.

Minutes Control group (ng / ml) Neck (ng / ml) 0 5.588 9.542 6.195 10 3.882 6.021 5.344 20 4.236 8.803 11.282 30 7.265 28.054 15.779 45 4.448 5.326 5.651 60 3.015 3.535 18.724

As shown in Table 2 above, after administering the neck extract of the present invention to the jugular vein of the rat at a concentration of 1 mg / kg of dry medicinal herbs using the animal, blood growth hormone was separated from the tail vein at regular intervals to measure the growth hormone of the plasma. As a result, compared with the control group, the drug increased about 2-3 times after 30 minutes of drug administration, and the growth hormone secretion promoting ability of the neck extract was measured within 1 hour ( FIG. 1 ).

Experimental Example 3 Oral Acute Toxicity in Rats

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, respectively, in the neck extract 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, and hematological and hematological examinations were performed.

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 throat extract of the present invention did not show a toxicity change up to 12.98 g extract / kg in rats, and the minimum lethal dose (LD ₅₀ ) of oral administration was determined to be a safe substance.

As described above, the neck extract of the present invention exhibits a very high growth hormone secretagogue activity, and since the neck is a raw material of the extract, which has been used for a long time, is harmless to humans and has good absorption rate, thereby treating various diseases related to growth hormone. And can be usefully used for prevention.

1 is a graph showing the growth hormone measured at regular intervals from the tail vein after administering the neck extract of the present invention to the jugular vein of the rat.

Claims (6)

It contains a tree extract as an active ingredient and promotes the secretion of growth hormone to treat or prevent diseases caused by growth hormone deficiency selected from the group consisting of arthritis, aging, Alzheimer's disease, dwarfism, obesity, osteoporosis and ulcers. Pharmaceutical compositions. The method of claim 1, wherein the extract is extracted by adding alcohol or a mixed solvent of alcohol and water to the barrel (Akebiae Caulis), or by hydrolyzing and neutralizing with an acid or base solution and then adding a mixed solvent of alcohol or alcohol and water Pharmaceutical composition, characterized in that. The method of claim 2, 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. Pharmaceutical compositions. The pharmaceutical composition according to claim 2, wherein the acid or base solution is at a concentration of 0.1 to 2 N. delete delete