KR20020046923A - Pharmaceutical Compositions and Health Foods for IGF-1 Release Comprising Extract from Phlomis umbrosa - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing a Phlomis umbrosa Turcz extract and having an insulin-like growth factor-1(IGF-1) secretion inducing effect is provided which can be mass-produced at low cost. A health food product containing the pharmaceutical composition can be administered orally to children and adults who are inactive in secretion of IGF-1. CONSTITUTION: Phlomis umbrosa Turcz is treated in hot water at 60 to 95deg.C and filtered with an ultrafiltration membrane having a molecular cut off of 30,000 to 100,000 to produce an extract. The titled composition contains: a pharmaceutically effective amount of the Phlomis umbrosa Turcz extract; a pharmaceutically acceptable carrier; and optionally a compound selected from the group consisting of calcium, arginine, lysine, carboxymethylcellulose and a mixture thereof.

Description

Pharmaceutical Compositions and Health Foods for IGF-1 Release Comprising Extract from Phlomis umbrosa

The present invention relates to a pharmaceutical composition comprising a rapid extract and inducing IGF-1 secretion, and more particularly, to the IGF-1 secretion comprising a pharmaceutically effective amount of a rapid extract prepared by a simple process from the fast-acting herbal medicine To pharmaceutical pharmaceutical compositions and health foods.

Insulin-like Growth Factor-1 (IGF-1) is a single-chain polypeptide consisting of 70 amino acids and is primarily secreted by the liver. IGF-1 exerts its function through the IGF-1 receptor. Many studies have been conducted on the in vivo function of IGF-1, and such studies have identified various functions other than promoting growth of IGF-1 (eg, promoting protein synthesis, reducing blood glucose levels, and promoting cell differentiation).

For example, Arsenijevic Y. et al, J. Neurosci. , 15; 21 (18): 7194 (2001) report that IGF-1 is essential for the proliferation of neuronal stem cells, and Cappola AR. et al., J. Clin. Endocrinol. Metab. , 86 (9): 4139 (2001) report a direct link between decreased muscle strength and decreased motor activity and decreased in vivo concentrations of IGF-1 in older women.

In addition, reports that skeletal muscle regeneration can be accomplished via IGF-1 have been reported by Shiotani A. et al., Gene Ther. 8 (14): 1043 (2001), and it has been reported that the bioavailability of IGF-1 is significantly reduced after ethanol overdose (Rojdmark S. et al., J. Endocrinol. Invest. , 24 (7): 476 (2001). On the other hand, it has been reported that IGF-1 plays an important role in cell proliferation (Singleton JR. Et al., Neurobiol. Dis. 8 (4): 541 (2001)), and diabetic patients (types 1 and 2). It has also been reported that IGF-1 has therapeutic effects in the form (Thraikill KM, Diabetes Technol Ther., 2 (1): 69-80 (2000)).

Seck T. et al. Found that low blood levels of IGF-1 were directly associated with femoral decline in postmenopausal women ( Clin. Endocrinol. , 55 (1): 101 (2001)), Conti. E. et al. Reported a rapid decrease in IGF-1 in the acute stage of myocardial infarction ( J. Am. Coll. Cardiol. , 38 (1): 26 (2001)).

Various reports have shown that IGF-1 has cellular protection against ischemia-reperfusion injury in various organs. Nakao Y. et al., J. Thorac Cardiovasc. Surg. , 122 (1): 136 (2001) report that IGF-1 has the ability to inhibit neuronal cell death. In addition, IGF-1 deficiency leads to a lack of postnatal growth, delayed mental development, microcephaly and insensitivity of sensory nerves, and in patients with these symptoms, growth hormone is normally secreted and signaling by growth hormone is normally achieved. However, it has been reported that IGF-1 is not produced locally or systemically (Woods KA. Et al., N. Engl. J. Med. , 335: 1363 (1996)). On the other hand, administration of IGF-1 to patients deficient in IGF-1 has been reported to improve gymnastics, insulin sensitivity, bone mineral density and length growth (KA Woods. Et al., J. Clin. Endocri. Met., 85: 1407 (2000).

See also Blum et al. ( J. Clin. Endocrinol. Metab. 76: 1610-1616 (1993)) reports that children with low levels of IGF-1 and IGF binding protein 3 (IGFBP-3) in the blood are short and tall Older children reported high levels of IGF-1. In another paper, long-term treatment of IGF-1 has been reported to be effective in increasing key growth rates (Ranke et al., Horm. Res. 44: 253-264 (1995)). It was found to be effective in the treatment of patients lacking growth hormone.

As described above, IGF-1 functions in vivo in a variety of functions, acts cooperatively or separately with the growth hormone, and in vivo by the growth hormone signal or individually. It can be seen that it has various functions.

Therefore, there is a need for a substance capable of promoting the production of IGF-1 in vivo.

MEANS TO SOLVE THE PROBLEM As a result of earnestly researching in order to solve the requirements of the said art, the present invention shows that the rapid extract which used conventionally as a herbal medicine exhibits the effect of inducing secretion of IGF-1 which exhibits the above-mentioned various functions, and the pharmaceutical containing a rapid extract And by confirming that the food is harmless to humans, the present invention has been completed.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for inducing IGF-1 secretion.

Another object of the present invention is to provide a health food for enhancing physiological activity having the purpose of treating and preventing various growth and physiological diseases caused by aging.

Still another object of the present invention is to provide a method for preparing a component having IGF-1 secretion inducing activity from the immediate organ.

1 is a graph showing a change in the concentration of IGF-1 in blood by the fast extract of the present invention,

Figure 2 is a graph showing the length growth of the thigh and tibia according to the long-term administration of the fast extract of the present invention, and

Figure 3 is a graph showing the results for the femoral portion of the results of FIG.

According to one aspect of the present invention, the present invention provides a pharmaceutical-effective amount of the fast-acting extract obtained by (a) extracting by heating the hot water and then filtering the hot-water extract using an ultrafiltration membrane having a molecular weight cutoff of 30,000-100,000; And (b) provides a pharmaceutical composition for inducing IGF-1 (Insulin like Growth Factor-1) secretion comprising a pharmaceutically acceptable carrier.

The pharmaceutical composition of the present invention which induces the above-described IGF-1 secretion may be administered to a subject having reduced in vivo production of IGF-1 to improve various symptoms and diseases. For example, it can improve gymnastics, insulin sensitivity, bone mineral density and length growth, and also inhibit mental retardation, microcephaly, insensitivity of sensory nerves and femoral decline in postmenopausal women, and inhibit the proliferation of nerve cells. It can promote and can suppress the decrease of muscle strength and the decrease of exercise ability in an elderly woman. And it can promote growth for children and adolescents whose growth plates are not closed but who are slow or lacking.

The candidate substance that can be used as a secretion inducing agent of IGF-1 having such various functions was used as a herbal medicine in the present invention, because the herbal medicine is a natural substance and has been used as a medicine in the past and its stability has been proved. Among the medicinal herbs investigated, fasting showed the best effect of inducing IGF-1 secretion.

On the other hand, the process of separating the physiologically active substances from natural products and separating them into meaningful purity should be accompanied by effective process development research. In particular, there are many considerations in terms of economics of production methods, maintenance and verification of the physiological activity of manufactured materials. Therefore, the present inventors have developed and confirmed a method for producing a large amount of extracts of IGF-1 secretion inducing components from Sokan, a natural herbal medicine, without loss of efficacy. In particular, in the case of fastening, since the material itself is expensive, the economic aspect is a very important consideration in the method of separating the bioactive material from the fastening.

Phlomis umbrosa Turez, which is the most important material in the present invention, is a perennial herb of 1 m high and is a nectar and a plant whose huge round roots are opened like sweet potatoes about five. The traditional pharmacological effect of fasting is the protection of the liver and kidneys (http://www.nanumy.co.kr/folk-remedy/documents/ic5.5-11.html). The present inventors confirmed that the fast extract having such efficacy can promote growth by inducing IGF-1 secretion in vivo.

In the pharmaceutical composition of the present invention, the extract of Sokdan is an extract obtained by extracting Sokdan by hot water heating and then filtering with an ultrafiltration membrane. The solvent used at the time of extraction by heating is water, which is intended to ensure that the resulting composition is not harmful to the human body, and an organic solvent such as methanol is not used in the present invention.

On the other hand, the heating is preferably performed at 60 ° C-95 ° C, and when the temperature is 60 ° C or less, the active ingredient is not well extracted from the rapid stage, and when it exceeds 95 ° C, destruction of the active ingredient may occur rapidly. Heating temperature becomes like this. More preferably, it is 80 degreeC-90 degreeC. The heat-extracted fast extract is then decanted, and then the precipitate is removed by centrifugation or filter paper.

The clear extract from which the precipitate is removed is then separated according to the molecular weight to obtain an extract of low molecular weight. Separation according to the molecular weight that can be used in the present invention is carried out by an ultrafiltration method, which is selected from the various methods in consideration of the yield, convenience and economy.

In the present invention, the ultrafiltration membrane used for ultrafiltration is preferably molecular weight cut off of 30,000-100,000, and if MWCO is less than 30,000, it takes a long time to separate the extract. There is a problem in economics, when using a membrane of more than 100,000 MWCO there is a problem that the relative purity of the extract of the present invention is finally lowered because the separation of the bioactive component and the macromolecular substance is lowered. As for the ultrafiltration membrane in this invention, what uses MWCO 50,000-100,000 more preferably.

In the present invention, more preferably, the concentration step is performed following the ultrafiltration process. By the concentration step, an extract containing a high concentration of pharmacological components is prepared. Concentration can be carried out in a variety of methods commonly used in the art, for example, by reduced pressure heating. On the other hand, if necessary, the finally prepared extract may be powdered by drying under reduced pressure.

The above-described manufacturing method for preparing the composition of the present invention has the advantage that can be easily carried out, while separating the key components having a high purity, high yield and economically IGF-1 secretion action.

Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

The pharmaceutical composition of the present invention has been developed for the purpose of oral administration, and especially can be used for various purposes, such as adult-oriented weak constitution improvement and anti-aging, osteoporosis prevention, gymnastic improver.

The pharmaceutical composition of the present invention has pharmacological effects such as treatment of length growth retardation, promotion of length growth, improvement of bone mineral density (prevention and treatment of osteoporosis), inhibition of aging, improvement of gymnastics, treatment of femoral decline in women after menopause and neurons Includes protection of

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. On the other hand, the oral dosage of the pharmaceutical composition of the present invention is preferably 5-50 mg / kg (body weight) per day.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporating into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or an aqueous medium, or may be in the form of extracts, powders, granules, tablets or capsules, and may further include a dispersant or stabilizer.

The composition of the present invention may further comprise calcium, arginine, lysine and / or carboxymethyl cellulose, if necessary. The additionally included calcium is the main inorganic component of bone, arginine is preferably added because it promotes the secretion of growth hormone in the body, lysine is preferably added because it enhances the action of arginine, carboxymethyl Cellulose is preferably added because it inhibits the moisture content of the composite composition, thereby reducing the possibility of the growth of microorganisms and at the same time facilitates uniform mixing of various substances.

The additionally included components are preferably 65 to 80 parts by weight of calcium, 25 to 40 parts by weight of arginine, 5 to 20 parts by weight of lysine, and 2 to 8 parts by weight of carboxymethyl cellulose, based on 100 parts by weight of the fast extract.

The main advantage of the composition of the present invention is that it can fully exhibit the effect orally as can be clearly seen in the following examples.

The health food of the present invention comprising the fast extract of the present invention may include ingredients that are commonly added during food production. For example, when the beverage is prepared, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, and / or juice may be further included in addition to the fast extract of the present invention. In addition, when prepared in powder form, the fast-acting extract of the present invention is dried under reduced pressure and powdered, and then the essential amino acids such as lysine, arginine, ornithine, glycine and tryptophan, and components such as niacin, gamma hydroxy butylate, etc. May be additionally included.

The present invention comprises the steps of (a) extracting the dried fastening using hot water; And (b) it provides a method for producing a composition comprising an IGF-secretion-inducing substance comprising the step of filtering the extract obtained by using an ultrafiltration membrane having a molecular weight cut off of 30,000-100,000.

The hot water used in the extraction step is preferably water at 60 ° C-95 ° C. On the other hand, the molecular weight cutoff of the ultrafiltration membrane is preferably 50,000-100,000.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it is to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. Will be self-evident.

Example 1 Preparation of Low Molecular Weight Fast Extract Using Ultrafiltration Membrane

To 50 g of dried fastener purchased from a domestic medicinal herb, 1.5 L of distilled water was added and heated and extracted at 80-90 ° C. for 2 hours to obtain 0.7 L of an aqueous extract. 0.7 L of the obtained extract was stored at room temperature, and distilled water corresponding to half of the amount added during the first extraction was added to the remaining medicinal herbs, and the second extraction was heated and extracted at 80-90 ° C. for 2 hours. One liter of the extract obtained over the first and second phases was collected and heated with heat (90 ° C.) to concentrate the volume of the extract to the final 500 ml.

500 ml of the obtained solution was centrifuged at 3000 xg for 10 minutes in a tabletop centrifuge, and then the supernatant was collected using a stirred cell apparatus (Amicon, United States) to obtain a molecular weight of MWCO 50,000 (diameter 76 mm) or MWCO 100,000 membrane. Passed in At this time, the pressure of nitrogen added was fixed at 3 atmospheres.

On the other hand, if clogging occurred during the concentration of the extract was replaced with a new ultrafiltration membrane to continue the operation, the membrane was washed with a washing solution (0.1N caustic soda, 20% ethanol) and reused. The ultrafiltration membrane used can be used up to MWCO 100,000.

In order to calculate the extraction efficiency in each of the heating and extraction steps and the filtration and concentration steps by the ultrafiltration membrane, the extract obtained in each step was dried under reduced pressure (freeze dryer, Edwards USA) to obtain a powder, and the weight of the powder The extraction efficiency was calculated based on (Table 1).

division Weight (g) Extraction efficiency (%) Early herbs 50 100 Heating and extraction stage 19.5 39 MWCO 50,000 Filtration 16.7 33.54 MWCO 100,000 filtration 18.5 37

The extraction efficiency of the heating and extraction steps in Table 1 was slightly varied each time, and generally showed an extraction efficiency of 38% -45%. As can be seen in Table 1, it can be seen that the MWCO of the ultrafiltration membrane is 100,000 or less is appropriate.

Example 2: In vivo IGF-1 secretion induction effect of the fast extract of the present invention

In Example 1, 252 mg of the extract powder obtained by passing MWCO 50,000 was dissolved in 1 ml of distilled water to prepare a dosage solution.

The animals used in this example are Sprague Dawley male rats with an average body weight of about 300 g. The experimental animals were fasted for one day prior to administration of the extract of the present invention for the purpose of enhancing the responsiveness to the secretory stimulation of IGF-1. The number of animals was 6 for the control group and 7 for the experimental group.

Subsequently, 1000 μl of the dose was forced into the rat by using an oral syringe. Blood collection was taken directly from the heart before and at 2 hours post-dose, and the last time was taken after 10 hours. In addition, anesthetics that may affect the secretion pattern of IGF-1 were not used during the experiment.

Then, the quantification of IGF-1 in the collected sample was measured according to the provided protocol by purchasing an EIA (enzyme immunoassay) kit manufactured and sold by the American Diagnostic System Laboratory (DSL). The result is as shown in FIG.

As can be seen in Figure 1, the experimental group to which the fast-acting extract of the present invention was administered after 4 hours after administration significantly increased the concentration of IGF-1 in the blood and the control group. Therefore, it can be seen that the fast extract of the present invention increases the concentration of IGF-1 in the blood in animals. In addition, the elevated blood IGF-1 level can be confirmed to recover to its original state over time. The results showed that blood IGF-1 concentration reached its highest point 8 hours after the administration of the sample. Therefore, it is clear that the fast extract of the present invention induces the secretion of IGF-1.

Example 3: In vivo biological activity effect of long-term administration of the fast extract of the present invention

Extracted powder obtained by passing MWCO 50,000 in Example 1 was prepared by adding to the feed composition as shown in Table 2 below:

ingredient Content (per 15 g of feed) Fast extract 50 mg Seaweed calcium 37.5 mg Arginine 16.8 mg Lysine 5.6 mg Carboxy methyl cellulose (CMC) 2.25 mg Etc -

The animal used in this example is a 3 week old Sprague Dawley male rat. The feed was given to the experimental rats for 8 weeks, and the rats were sacrificed to extract femoral and shin bones, and the change in length growth was measured. The control group is a group of animals that are allowed to take the feed except the fast extract of the present invention. Experimental results are shown in FIGS. 2 and 3.

As can be seen in Figures 2 and 3, it can be seen that the length of the thigh and shin bone, which is an indicator of the growth of the length of the animal is increased by the extract of the present invention.

Preparation Example 1 Preparation of Health Food Drink

In Example 1, 80 ml of extract obtained by passing MWCO 50,000, 10 ml of liquid fructose, 0.5 ml of citric acid, and 9.5 ml of licorice extract were mixed to prepare a drink of the present invention.

As described in detail above, the present invention provides a pharmaceutical composition and health food comprising the IGF-1 secretion induction component. Since the pharmaceutical composition of the present invention includes a fast-acting extract that has been used as a herbal medicine as an active ingredient, it is stable to the human body, and the fast-acting extract containing IGF-1 secretion inducing components can be prepared in large quantities in a simple process. It is economically very useful and can significantly contribute to securing a large amount of IGF-1 secretion promoter raw materials that have not been solved in the prior art. In addition, the fast extract of the present invention can be administered orally in the form of a medicine or health food to children and adults with low secretion of IGF-1.

Claims (6)

A pharmaceutically effective amount of the fast extract obtained by extracting the fast extract by hydrothermal treatment and then filtering the hot water extract using an ultrafiltration membrane having a molecular weight cutoff of 30,000-100,000; And A pharmaceutical composition for inducing IGF-1 secretion comprising a pharmaceutically acceptable carrier. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises one selected from the group consisting of calcium, arginine, lysine, carboxymethyl cellulose, and mixtures thereof. A health food for inducing IGF-1 secretion comprising fast extract extract obtained by filtering fast extract by hot water treatment and then filtering the hot water extract using an ultrafiltration membrane having a molecular weight cutoff of 30,000-10,000. (a) extracting the dried fastener using hot water; And (b) The method of producing a composition comprising an IGF-secretion inducing substance comprising the step of filtering the extract obtained by using an ultrafiltration membrane having a molecular weight cutoff of 30,000-100,000. 5. The method of claim 4, wherein the hot water is water at 60 ° C.-95 ° C. 6. 5. The method according to claim 4, wherein the ultrafiltration membrane has a molecular weight cutoff of 50,000-100,000.