KR20050005633A - Lonicera japonica extract having activity of growth hormone secretagogue, its process method, and application - Google Patents

Abstract

PURPOSE: Provided are the Lonicera japonica extract having activity of growth hormone secretagogue, a method for preparing the same by extracting Lonicera japonica with water, alcohol or their mixture, and the application thereof to cosmetics, remedies and foods for enhancing growth hormone secretion. CONSTITUTION: The composition inducing and/or enhancing growth hormone secretion is characterized by containing, as an active ingredient, 0.0001-10 wt.% of the extract of Lonicera japonica. The extract contains at least one active ingredient selected from the group consisting of loniceroside A, B, C, hederagenin derivative, lonicerin, loganin and iridoid glycosides including vogeloside and Epi-vogeloside.

Description

Phosphorus extract with excellent growth hormone secretion activity, preparation method and use thereof {Lonicera japonica extract having activity of growth hormone secretagogue, its process method, and application}

The present invention relates to a method for producing an extract of Indonesia ( Lonicera japonica ), an extract obtained by the method and a composition comprising the same as an active ingredient, more specifically, by adding water, alcohol or a mixed solvent of alcohol and water It relates to a method of preparing the extract of Indongcho extract, Indongcho extract obtained by the above method and a pharmaceutical composition, cosmetic composition or health food composition comprising the same as an active ingredient.

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 concentration and decrease 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). In addition, the effects of sex hormones have been applied in various fields, such as improving immunity by stimulating hormone secretion (J. immunol. 2001, Mar, 15; 166 (6): 4195), and results that growth hormone improves bone density in adults. (Endocrine 2001 Feb; 14 (1): 63-6), reports that promoting growth hormone secretion improves the effects of sleep at bedtime (US Pat. No. 6,313,133), and reports a close relationship between hormones and heart disease (J. clin. endocrinol. metab. 2000. 86 (5 ), 1871-3), has been reported cholesterol-lowering, improved wrinkle removal, cellulite removal, kidney function, improve sexual performance, improve exercise capacity, such as blood pressure, improve memory. (growth young with HGH by Dr. Ronald Klatz)

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 deficiency of many types of pituitary hormone than in adults, 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 secretion. Ghrelin, a ligand, was effective when administered to growth hormone deficient mice ( Diabetes , 2001, 50 (2), 227-32), active growth hormone secretagogue MK-677, and a potent bone resorption inhibitor, 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 are 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 stimulate the growth hormone secretion (Korean Patent Registration 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), as the original extract, the casualty extract, the lily extract and the Giltyeong extract. Of a growth hormone secretagogue composition containing one or more extracts selected from the group consisting of (Korean Patent Application No. 1998-0053921), Method for extracting growth hormone secretion stimulating factor from natural herbal medicine and water-soluble fast extract (Korea Patent Application 1998-014589) and growth hormone powder of asperosaponin H1 (glycoalkaloids) There have been reported studies on stimulators (Republic of Korea Patent Application No. 1998-014590).

Indong is a semi-evergreen vine shrub of the Capriofoliaceae family that grows under the forests of the country's wild field, and the bud is called 'Gold Silver Coin' and the leaves and stems are called 'Indongdeung'. This is a plant that has been widely used in herbal medicine. The active ingredients contained in phosphorus and the like so far are known as lonerin (luteolin-7-rhamnoglucoside), caffeoylquinicacid, methyl caffeate, chlorogenic acid, iso-chlorogenic acid (iso flavonoids such as -chlorogenic acid; Biflavonides, such as ochnaflavon; Saponin is a fulvotomentoside A, α-hederin, sapindoside B and new material Loniceroside A identified by the present inventors, B (Korean Patent Publication No. 94-2267); Contains iridoid glycosides such as Loganin, Vogeloside, Epi-vogeloside, etc. Turned out to be. These sinus has been widely used in arthritis, bronchitis, asthma, etc., since the prosthesis is effective for all scabs (inflammation of the inside and the outside), and in recent years, pandemic flu, various purulent infections, moist fever It is known to be effective in restoring joint redness, swelling, hepatotoxicity, and recovery of decreased cellular immunity.

However, there are no published studies on the effects of growth hormone secretion on humans, although most of them are mentioned only in the herbal medicine or related literature. Patent 0163813 is effective in maximizing the extraction efficiency of herbal ingredients of phosphorus and using the phosphorus more scientifically to extract stems and leaves of phosphorus during the growth period with the highest content of active substance and optimizing the addition ratio of stems and leaves. The method of extracting the active ingredient was determined, and at the same time, the method of extracting the active ingredient having anti-inflammatory analgesic physiological activity with the most active composition by controlling the timing of extracting the phosphorus and the mixing ratio of the leaves and stems, and the extract containing the active ingredient There is a patent on the herbal composition, and Kang et al. (Application No. 1992-0011752) from the human patents, Kang et al., (I1995-0007924) on the isolation and effectiveness of bifurabonoids effective against anti-inflammatory, anti-allergic and antirheumatic A patent has been filed for the isolation and structure determination of a headraginin glycoside single material.

Hereinafter, the present invention will be described in detail.

The present invention relates to the effect of promoting the growth hormone secretion by extracting and purifying the active ingredient from the phosphorus. The phosphorus usable in the present invention is a hormone, which is used to extract the flowers, leaves, fruits, stems, and roots of the honeysuckle regardless of the season in which it is collected. It can be used as a material for secretory accelerators. In addition, the present invention is Lonieroside A (Lonieroside A), Lonieroside B, Lonieroside C, and hedragenin derivative (Lonicerin), Loganin (Loganin), etc. Contains herbal compositions that are contained.

Referring to the present invention in more detail as follows.

The present invention is a method for extracting and purifying an active ingredient having excellent growth hormone secretion physiological activity from stems and leaves of phosphorus (or may be called indongcho, but the same meaning). By using each of them collected at a specific time, the active ingredient content can be maximized.

In the present invention, the mixed herbal leaves and stems of the herbal medicines extracted with water, and extracted by reflux for about 5 hours by adding water of 8 to 12 times the weight of the herbal medicines, filtered and collected the filtrate, and the residue 3 again the weight of the herbal medicines weight The water extraction efficiency is increased by adding ˜7 times of water, heating and filtration to mix the filtrate with the previous filtrate.

The herbal residue was removed from the water extract by centrifugation and filtration, and then the separation solution was adsorbed onto HP-20 as an adsorbent to separate the bioactive component fraction. The column on which the physiologically active component was adsorbed was sufficiently washed with water and eluted with ethanol. The eluted solution was concentrated under reduced pressure at 50 degrees and dried in vacuo.

Loniceroside A, B, C and hedragenin analyzing growth hormone secretion promoting physiologically active substances of standardized fractions extracted from the leaves and stems of phosphorus by the method according to the present invention as described above In addition to derivatives, known ingredients such as Loganin and Loginerin are included. Therefore, when the herbal composition is prepared and used as a powder extract obtained by the extraction and purification method according to the present invention, it may be usefully used as a growth hormone secretagogue.

The herbal composition prepared from the powder extract according to the present invention may be formulated in a conventional manner, and then administered in the form of tablets, soft capsules, injections, etc., and the bases, for example, lactose (Lactose), fine Base ingredients such as crystalline cellulose (Magnesium Stearate) and phosphorus powder extract can be used in an appropriate mixture. Preferably, when used in a ratio of 1: 1.5 to 1.5: 1, it is superior to conventional ones. It exhibits a growth hormone secretagogue activity.

In particular, when the herbal composition of the present invention is administered to the human body, there is little concern about side effects compared to other synthetic drugs because it is a natural extract. It has been demonstrated in animal model experiments for gastrointestinal disorders and ulcerative side effects. That is, the phosphorus extract prepared in the present invention was excellent in promoting the growth hormone secretion effect (see Experimental Example), and the general phosphorus extract was found to be completely nontoxic as a result of acute toxicity test in mice and rats. . (LD ₅₀ = 5 g ext's / Kg or more, Journal of Pharmacognosy Vol. 25, No. 4, 1994).

As described above, according to the present invention, by extracting and purifying the active physiologically active ingredient from the phosphorus, it is possible to administer the phosphorus extract, which was conventionally used only in the form of medicinal herbs, in the form of powders, ointments, tablets, injections, etc., which are convenient to administer. It has an effect. Accordingly, the present inventors have completed the present invention by finding out that the physiological activity of the extract of Indongcho extract increases the production of growth hormone by acting as a growth hormone secretagogue.

It is an object of the present invention to provide a method for preparing Indongcho extract having growth hormone secretagogue activity. On the other hand, an object of the present invention is to provide a composition of various forms, such as the extract of Indongcho obtained by the above method and a pharmaceutical composition, a cosmetic composition and a health food composition containing the same as an active ingredient.

In order to achieve the above object, the present invention provides a method for producing Indongcho extract. In addition, the present invention provides an extract of Indongcho having growth hormone secretagogue activity prepared by the above method. In addition, the present invention provides a pharmaceutical composition, cosmetic composition, and health food composition for promoting growth hormone secretion containing the extract of Indongcho as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a method for preparing Indongcho extract. Specifically, the present invention provides a method of preparing the phosphorus extract by adding water, alcohol, or a mixed solvent of alcohol and water and immersing at 5 to 100 ° C. In the above water, distilled water, purified water, or general drinkable water and alcohol may be selected from the group consisting of 10 to 100% ethyl alcohol, 10 to 100% methyl alcohol and alcohol. . Extraction temperature in the extraction step is 0 to 120 ° C, preferably 30 to 100 ° C. Range, the extraction time is in the range of 1 minute to 24 hours, preferably 1 hour to 5 hours, and the pressure at extraction is in the range of 0 to 1.5 psi.

The present invention was used to adsorb the active ingredient to the adsorption resin HP-20 is extracted from the Indongcho extract extracted by the above method. Specific examples of the adsorbent resin include acrylic resins such as acrylic esters, polymethacrylates, and polystyrene aromatic resins. The extract is passed through a column packed with an adsorptive resin to adsorb a substance having growth hormone secretion physiological activity in the extract onto the resin. The adsorption process is carried out by flowing a non-polar or slightly polar adsorbent resin at a flow rate of 0 to 100 占 at a flow rate of, for example, 1 to 4 l / min.

Adsorbed to the resin by passing all the extract of Indongcho containing the substance having growth hormone secretion physiological activity, and then flowing distilled water sufficiently to wash the resin, and then passing the alcoholic solvent such as ethanol through the column. It is possible to recover a substance having a growth hormone secretion physiological activity. The recovered alcohol-containing eluate having growth hormone secretion physiological activity was concentrated according to a conventional method. The concentration process can be carried out by, for example, concentration under reduced pressure at a temperature of 40 to 80 占 폚 to obtain a concentrate, followed by drying under reduced pressure at 30 to 80 占 폚.

In addition, the present invention provides an extract of Indongcho having growth hormone secretagogue activity prepared by the above method.

The present inventors confirmed whether the extract of Indongcho 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. Indongchocho extract of the present invention was added to the pituitary cells isolated from the above, and in the comparison group, 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 extract of Indongcho 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.

In addition, growth hormone secretion experiments using animal models showed that Indongcho extract strongly increased growth hormone in vitro as well as in vivo .

The present invention also provides a pharmaceutical composition for promoting growth hormone secretion containing the extract of Indongcho as an active ingredient.

The pharmaceutical composition for promoting growth hormone secretion of the present invention contains the extract of Indongcho as an active ingredient. The Inchocho 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 extract of Indongcho of the present invention can be administered in various oral and parenteral formulations during 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 such solid preparations contain at least one excipient such as starch, calcium carbonate, sucrose, lactose and gelatin in the extract of Indongcho. 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 Indongcho extract is concentration dependent, but preferably 0.1 to 1 g / kg, more preferably 0.4 to 200 mg / kg, and may be administered 1-6 times a day.

The present invention also provides a health food composition for promoting growth hormone secretion containing Indongcho 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 an extract of Indongcho to the general food, and the Indongcho 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 the phosphorus extract of the present invention is used as a food additive, the phosphorus extract may be added as it is or used together with other food or food ingredients, and may be appropriately 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 extract of Indongcho 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 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 is preferably to contain the extract of Indongcho within the range of toxicity measured when used in the pharmaceutical composition.

There is no particular limitation on the kind of food. Examples of foods to which the extract may be added include meat, sausage, bread, chocolate, candy, snacks, confectionary, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks Alcoholic beverages and vitamin complexes. Specifically, the health food containing the extract of Indongcho may include health foods and favorite products such as juice, tea, jelly, and juice made mainly from Indongcho, and include folk remedies for the purpose of edema, nephritis, urethritis, and the like. 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 Indongcho extract 1

50 g of Indongcho dry powder were added to 500 ml of methanol 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 the extract of Indongcho of the present invention.

Example 2 Preparation of Indongcho Extract 2

50 g of Indongcho dried powder was added to 500 ml of distilled water, and extracted at 100 ° C. for 3 hours. After extracting for 3 hours to remove insoluble solids by filtration and centrifugation, the supernatant was concentrated under reduced pressure to freeze-dried to prepare the extract of Indongcho of the present invention.

Example 3 Purification of Indongcho Extract

50 g of Indongcho dried powder was added to 500 ml of distilled water, and extracted at 100 ° C. for 3 hours. After extracting for 3 hours, insoluble solids were removed by filtration and centrifugation, and the supernatant was flowed to HP-20 resin-adsorbed column (trade name DIAION) at a flow rate of 20 ml / min to promote growth hormone secretion in the concentrate. It was allowed to adsorb.

After distilled water was sufficiently flowed into the column to wash the resin, ethanol (95%) was flowed to recover the fraction containing the growth hormone secretagogue component adsorbed on the resin. The recovered ethanol eluate was concentrated under reduced pressure at a temperature of 60 ° C. to obtain a concentrate, and the concentrate was dried under reduced pressure at 65 ° C. to obtain an active fraction of 2.5%.

Experimental Example 1. Growth Hormone Secretion Activity Test Using Indongcho Extract

1. Pituitary Cell Culture

Drainage of the pituitary gland under aseptic manipulation from Sprague-Dawley rats at 4-5 weeks of age, with collagenase (Gibco co., Grand Island, NY) and hyaluronidase (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. Incubation at 37 ° C., 5% CO 2).

2. Growth hormone secretion promoting activity experiment

The pituitary cells isolated above 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 125I-labeled growth hormone and the test substance stored above was different from 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 125I was measured from the precipitation. The results are shown in Table 1 below.

TABLE 1

Added concentration Growth hormone concentration (ng / ml) Control 0 24.2 Comparison 300 nM 36.5 500 nM 54.8 1,000 nM 137.6 Example 3 1mg Medicine / ml 312.5

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

As a result, as can be seen in Table 1 , the extract of Indongcho 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 in the comparison group. It was confirmed.

Experimental Example 2. Growth Hormone-induced Experiments in Rats

Eight weeks-old Sprague-Dawley rats were anesthetized at a concentration of 50 mg / kg of pentobarbital, and Example 3 was dissolved in physiological saline at a concentration of 50 mg / kg, and orally administered to rats. 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. 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 secondary antibody to the precipitate¹²⁵The radioactivity of I was measured and the corresponding concentration was calculated using a standard curve.

TABLE 2

Hours (hrs) Control group (ng / ml) Example 3 (ng / ml) 0 6.527 7.623 6.358 2 5.483 8.257 7.836 4 4.859 18.369 21.593 6 6.245 31.475 28.485 8 5.483 26.385 19.368 10 4.236 15.735 12.639

As shown in Table 2 , the rats were orally administered to rats of Example 3 of the present invention at a concentration of 50 mg / kg and collected at regular time intervals from the tail vein to measure growth hormone of the separated plasma. Compared to 6 hours after drug administration, the increase was about 4 times.

Example 4 Manufacturing method of injection solution

Injection solution containing 10 mg of the active ingredient was prepared by the following method. 10 g of Indongcho extract was dissolved in 1,000 ml of saline solution and used as an injection by 0.2um sterile filtration.

The components of the injection solution are as follows.

Example 3 10 g

1,000 ml of saline

Example 5 Tablet manufacturing method

Tablets containing the active ingredient are prepared by the following method. A tablet was prepared by mixing 600 g of Indongcho extract, 240 g of arginine, 140 g of crystalline cellulose, 10 g of magnesium stearate, and hydroxypropylmethylcellulose.

The components of the tablet are as follows.

600 g of Indongcho extract

Arginine 240 g

140 g of crystalline cellulose

10 g of magnesium stearate

10 g of hydroxypropyl methyl cellulose

Example 6 Application of Indongcho Extract to Milk

Milk ----------------------- 100g

Example 2 --------------------- 1g

Example 7 Application of Indongcho Extract to Orange Juice

Liquid fructose -------------------- 5%

Polydextrose --------------- 1%

Citric acid --------------------- 5%

Vitamin C -------------------- 0.02%

Example 9 -------------------- 2%

Orange Juice Concentrate ------------- 25%

Water ------------------------ 67%

Example 8 Application of Honeysuckle Extract to Cosmetic Lotion

1,3-butylene glycol --------------- 5%

glycerine ----------------------- 5%

EDTA-2Na --------------------- 0.02%

trimethylglycine ----------------- 2.0%

cetanol ------------------------ 1.0%

glyceryl monostearate emulsifier --- 1.0%

polysorbate 60 ------------------ 1.2%

sorbitan sesquioleate ------------- 0.3%

cetyl 2-ethyl-hexaoate ----------- 4.0%

squalane ----------------------- 5.0%

dimethicone --------------------- 0.3%

glyceryl stearate ----------------- 0.5%

carbomer ----------------------- 0.15%

triethanoamine ------------------- 0.5%

imidazolidinyl urea ---------------- 0.2%

Example 3 ------------------------- 2.0%

Purified Water -------------------------- 71.8%

Example 9. Indongcho extract Application to cosmetic skin

1,3-butylene glycol --------------- 4.0%

dipropylene glycol ---------------- 5.0%

EDTA-2Na ---------------------- 0.02%

octyldodeceth-16 ----------------- 0.3%

PEG60 hydrogenate castor oil ------- 0.2%

Example 3 ------------------------- 0.5%

Purified Water -------------------------- 90%

Example 10 . Manufacture of beverages

Calcium Citrate 50 mg

Citric acid 5 mg

Cortinamide 10 mg

Riboflavin Sodium Hydrochloride 3 mg

Pyridoxine hydrochloride 2 mg

Arginine 30 mg

Example 2 100 mg

200 ml of water

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

As described above, Indongcho extract of the present invention exhibits a very high growth hormone secretagogue activity, and the raw material of this extract is harmless to the human body has been used for a long time and good absorption rate, so the treatment of various diseases related to growth hormone And can be usefully used for prevention.

Claims (23)

Growth hormone secretion induction and / or promotion composition comprising Lonicera japonica extract as an active ingredient The composition of claim 1, wherein the extract of Indongcho is a extract of flowers (gold) and fruit, roots and / or leaves and stems (dongdong, etc.). The method of claim 1, wherein the extract of Indongcho is Loniceroside A, Lonieroside B Loneroside C, hedragenin, Lonicerin, Loganin and Vogelolo Composition comprising at least one active ingredient selected from the group consisting of Iridoid glycosides, such as Vogeloside, Epi-vogeloside The composition of claim 1 or 2, wherein the extract of Indongcho is immersed and extracted. The composition of claim 1 or 2, wherein the phosphorus extract is extracted with water, an alcohol, or a mixed solvent of alcohol and water. 6. A composition according to any one of claims 1 to 5, wherein said composition has dwarfness therapeutic activity. The composition according to any one of claims 1 to 5, wherein the composition has menopausal disorder improvement and / or aging delaying activity. 6. A composition according to any one of claims 1 to 5, comprising a pharmaceutically acceptable carrier or excipient. The composition of claim 8, wherein the pharmaceutically acceptable carrier or excipient is at least one selected from base components such as lactose, microcrystalline cellulose, magnesium stearate, and the like. The composition according to claim 8, wherein the mixing ratio of the pharmaceutically acceptable carrier or excipient and the phosphorus extract is 1: 1.5 to 1.5: 1. The composition according to any one of claims 1 to 5, comprising a carrier or excipient which is acceptable for the preparation of cosmetic compositions. 6. The composition of claim 1, comprising a pharmaceutically acceptable carrier or excipient in the form of an edible beverage or food. 7. 13. The composition according to claim 12, wherein the content of the extract of Indongcho is 0.0001 to 10% by weight. The growth hormone secretagogue according to any one of claims 1 to 5, comprising the composition as an active ingredient and optionally comprising one or more pharmaceutically acceptable carriers or excipients. The formulation of claim 14 further comprising an antacid. 15. The formulation according to claim 14, wherein the formulation is a tablet, powder, hard or soft capsule, suspension, injectable formulation, emulsion, suppository, unit dosage form or parenteral administration for parenteral administration. 17. The formulation according to claim 14, wherein the content of the active ingredient is administered in the range of 25-24,000 mg / day per 1 kg of adult body weight. a) obtaining crude crude by refluxing the cauliflower with a water-soluble solvent; b) optionally, concentrating or filtering the extract; c) adsorbing onto HP-20 as an adsorbent; and d) adsorbing the column onto which the active ingredient is adsorbed. Washing with water and eluting the active ingredient with a water-soluble solvent; and e) optionally drying under reduced pressure at a suitable temperature under vacuum drying. 19. The method according to claim 18, further comprising a purification step by an anion exchange chromatography and a dialysis step before step c). 19. The process of claim 18, wherein the water soluble solvent for extraction is water, lower alcohols or mixtures thereof. The method of claim 20, wherein the lower alcohol is ethanol, methanol, isophoropil alcohol, propanol, acetone. 22. A phosphorus (Indongcho) extract prepared by the method of any one of claims 18 to 21. The method of claim 22, wherein the extract is Loniceroside (A), (B), (C), hegragenin derivatives, Loganin (Longerin), Lonicerin, (Vogeloside), Extract characterized in that it comprises at least one active ingredient selected from the group consisting of Iridoid glycosides such as Epi-vogeloside (Epi-vogeloside)