KR101330044B1 - Composition for improving obesity including compounds from cordyceps militaris having anti-obesity activity - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for improving obesity, including a Militaris cordyceps sinensis derived compound having an anti-obesity effect.
The pharmaceutical composition for improving obesity of the present invention comprises a compound consisting of two or more selected from Militaris cordyceps derived cordycepin, guanosine and tryptophan.
According to the present invention, there is provided a pharmaceutical composition for improving obesity, which is harmless to the human body, while having an excellent anti-obesity effect and without causing side effects and toxicity.

Description

COMPOSITION FOR IMPROVING OBESITY INCLUDING COMPOUNDS FROM CORDYCEPS MILITARIS HAVING ANTI-OBESITY ACTIVITY}

The present invention relates to a pharmaceutical composition for improving obesity, and particularly relates to a pharmaceutical composition for improving obesity, including a Militaris cordyceps sinensis-derived compound having an excellent anti-obesity effect.

Current methods used to treat obesity can be divided into methods of correcting lifestyles such as diet therapy, exercise therapy, behavioral therapy, drug treatment and surgical treatment.

Correcting lifestyles is not only easy, but weight loss is limited and requires medication along with lifestyle correction.

Although the use of drugs for the treatment of obesity has been tried for a long time, obesity treatment drugs targeting many substances that have been found to be involved in the regulation of energy homeostasis of the human body has limited clinical applications due to side effects.

There are currently two types of obesity drugs approved by the US FDA for long-term use: sibutramine (sibutramine), which inhibits reuptake of norepinephrine and serotonin, and lipolytic secretion from the pancreas and digestive system. There is an orlistat that suppresses enzymes and has an anti-obesity effect.

However, in the case of sibutramine, side effects such as increased blood pressure, insomnia, dry mouth, dizziness, and the like often occur, and there is a disadvantage that it can not be used in patients with cardiovascular disease and uncontrolled hypertension. In addition, orlistat has side effects such as diarrhea, fatty stools, and urinary incontinence, and the use of the drug is limited because the effect of the drug is less pronounced when the fat intake is less than that of Westerners such as Koreans.

Therefore, recently, studies on how to safely treat obesity using natural products derived from biologically active substances have been actively conducted.

That is, Korean Patent Publication No. 10-0799116 (pharmaceutical composition for the treatment and prevention of obesity containing cordycepin) discloses a composition which contains the codeispin derived from Militaris cordyceps as a compound exhibiting an effect on the treatment of obesity. have.

However, the preceding data as one of the compounds derived from Miltalis Cordyceps Sinensis have the effect of alleviating and improving the symptoms of obesity, but the effect is inadequate and can still be safely administered without any side effects and toxicity. There is still a need for it.

Korean Patent Publication No. 10-0799116 "Pharmaceutical composition for the treatment and prevention of obesity containing cordycepin"

An object of the present invention is to provide a pharmaceutical composition for improving the harmless obesity of the human body having excellent anti-obesity effect and no side effects and toxicity.

Technical problems to be achieved by the present invention are not limited to the above-mentioned technical problems, and other technical problems not mentioned above may be clearly understood by those skilled in the art from the following description. There will be.

Obesity improvement pharmaceutical composition of the present invention for achieving the above object is composed of a compound consisting of two or more selected from cordycepin (cordycepin), guanosine (tryptophan) derived from Militaris Cordyceps sinensis .

The compound is characterized by being separated by extracting Militaris Cordyceps sinensis in a solvent and fractionating it with butanol.

The solvent is used by mixing one or two selected from water or ethanol, in particular, it is preferable to use a mixture made of water and ethanol in a 1: 1 volume ratio as the solvent.

The compound is characterized in that the two selected from cordycepin (cordycepin), guanosine (guanosine), tryptophan (tryptophan) 45:55 weight ratio or 50:50 weight ratio.

The compound is characterized by consisting of cordycepin (cordycepin), guanosine (guanosine), tryptophan (tryptophan) in a weight ratio of 20 to 30:10 to 20:50 to 60.

The pharmaceutical composition is characterized by inhibiting adipocyte differentiation.

As described above, by providing a pharmaceutical composition for improving the harmless obesity of the human body having excellent anti-obesity effect and without the risk of side effects and toxicity, by providing the health functional foods with functional and safety to the people It can contribute to improving public health.

1 is a diagram showing the fat differentiation (producing) inhibitory activity of Militaris cordyceps sinensis.
Figure 2 is a diagram showing the separation of fat differentiation (producing) inhibitor of Militaris cordyceps.
3 is a diagram showing the effect of inhibiting the differentiation (production) of the extract of Militaris cordyceps sinensis.
Figure 4 is a diagram showing the separation of compounds of the Militaris cordyceps butanol fraction.
5 is a diagram showing the structural analysis of the compounds separated in FIG.
Compound 1: cordycepin
Compound 2: Guanosine
Compound 3: tryptophan
6 is a diagram showing the effect of inhibiting fat differentiation (production) of the compound isolated in FIG.
FIG. 7 is a diagram showing the effect of inhibiting fat differentiation (production) according to the combined administration of the compounds separated from FIG. 4.
8 is a view showing the content of each compound according to the extraction solvent of Militaris cordyceps.
9 is a diagram showing the cytotoxicity of the compound isolated in FIG.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described in detail with reference to the accompanying drawings.

According to the plant taxonomy, Cordyceps belongs to Ascomycota, Clavicipitales, Clavicipitaceae, and three genera of Cordyceps, Podonectria and Torrubiella exist. Among them, the genus Cordyceps is representative.

Militaris cordyceps, belonging to the genus Cordyceps, is distributed in Korea, Japan and the United States and is also called pupa mushroom. Occurs in insect bodies in the land of mixed forests from summer to autumn. Parasitic on the pupa of lepidoptera insects. Fruiting bodies occur at the head of the chrysalis body, club-shaped, sack is round columnar, slightly curved, orange, but the head is fusiform and vivid orange. The pizzeria is embedded in the epidermis of the head, and there are thread-shaped spores in the inner scapula, and when mature, they split again to form secondary spores.

Militaris Cordyceps sinensis is known to have various effects such as anti-inflammatory, antibacterial, anti-tumor, and immune-enhancing effects such as protection, anti-aging, disease suppression and alleviation, but obesity prevention or treatment through inhibition of fat accumulation No effect was previously known of the present invention. The present inventors have found that the extract Millitaris cordyceps sinensis inhibits fat accumulation by decreasing blood triglycerides and increasing blood HDL-cholesterol.

The present invention is to provide a pharmaceutical composition exhibiting excellent anti-obesity activity, and is isolated from the various components of the Militaris Cordyceps sinensis, such as a substance that exhibits substantially inhibiting the differentiation activity, without showing toxicity to fat cells The composition of the pharmaceutical composition showing the best effect was found to include two or more selected from cordycepin, guanosine, and tryptophan, so that the fat is superior to the administration of the compounds alone. It exhibits a cell differentiation inhibitory ability to provide a pharmaceutical composition for improving obesity having excellent anti-obesity activity.

In one embodiment of the present invention, the compounds are extracted by putting the Militaris Cordyceps sinensis in a solvent and fractionated with butanol, preferably the solvent is a hydrophilic organic solvent such as water or ethanol or a mixed solvent thereof After extraction, it is preferable to separate by fractionation with butanol, and more preferably, a mixed solvent in which ethanol and water are mixed in a 1: 1 ratio so that all of the compounds (cordicepin, guanosine, tritopane) are uniformly extracted. It is good to use

The fraction thus formed contains compounds of cordycepin, guanosine, and tryptophan, and these fractions are prepared as a dried product, a crude product, or a purified product obtained by dilution, concentration or drying.

Compounds thus produced, namely cordycepin, guanosine, and tryptophan, are composed of two selected from the above compounds to have excellent anti-obesity activity, but higher fat cells than those administered alone. In order to have a storage inhibitory capacity, it is preferable that the ratio is composed of 45:55 weight ratio or 50:50 weight ratio.

At this time, the 45:55 weight ratio is composed of cordycepin, guanosine (guanosine) when one selected from 45% by weight of the remaining tryptophan (tryptophan) consists of 55% by weight, guanosine (guanosine) , 45% by weight of the selected one of the tryptophan (tryptophan) consists of the remaining cordycepin (55% by weight), 45% by weight of one selected from cordycepin (cordycepin), tryptophan (tryptophan) Guanosine is composed of 55% by weight.

Preferably, it consists of three types of cordycepin, guanosine, and tryptophan, and 20 ~ 30: 10 in order to have better fat cell accumulation inhibitory effect than two compounds. 20 to 50 to 60 weight ratio is good.

Such pharmaceutical compositions may further include a pharmaceutically acceptable carrier, which is commonly used in the preparation of formulations, 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 It includes, but is not limited to.

In addition to the above components, the pharmaceutical composition of the present invention may further include excipients such as lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like.

The pharmaceutical compositions of the present invention may be administered parenterally, with subcutaneous infusion or topical administration via the skin (transdermal administration) being preferred.

Suitable dosages of the pharmaceutical compositions of the present invention may be prescribed in various ways 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 0.001 to 100 mg / kg (body weight) per day.

The pharmaceutical compositions of the invention may be formulated in unit dosage form by formulating with any one or more of pharmaceutically acceptable carriers or excipients, according to methods which may be readily practiced by those skilled in the art. It can be prepared by or incorporated into a multi-dose container.

The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples, but the scope of the present invention is not limited thereto.

Example 1 Isolation and Identification of Active Substances

1) Induction of allergic cell differentiation

Differentiation-inducing substances IBMX (Isobutylmethylxanthine), dexamethasone, and insulin (Insulin) were added to adipocytes (3T3-L1) to induce differentiation into adipocytes.

2) fat cell production measurement

The cells were measured by differentiation (diogenesis) into fat cells by Oil Red O staining, and confirmed the ability to inhibit adipocyte differentiation (generation) when treated with Militaris cordyceps itself (FIG. 1).

3) Extraction and fractionation of Militaris cordyceps

Ethanol extracts of Militaris cordyceps were treated with nucleic acid, chloroform, butanol, and separated into nucleic acid fractions, chloroform fractions, butanol fractions, and water fractions (FIG. 2).

The fractions isolated as described above were confirmed to have the highest activity in the butanol fraction, which confirmed the inhibitory effect of adipocytes by inducing adipose differentiation and measuring the production of adipocytes according to the methods of 1) and 2) above ( 3).

4) Compound Separation of Butanol Fraction

Thus, three compounds were separated by chromatography in the butanol fraction (FIG. 4).

5) Butanol fraction compound structure analysis

Compounds separated as described above were identified and identified using NMR, MS, and the like.

As a result, as shown in FIG. 5, compound (1) was identified as cordycepin, compound (2) as guanosine, and compound (3) as tryptophan.

<Example 2> Preparation of the pharmaceutical composition for improving obesity, including Militaris cordyceps sinensis derived compound having an anti-obesity effect of the present invention

3 μg / mL of the composition containing a mixture of the identified compounds cordycepin, guanosine and tryptophan in a 1: 1: 1 weight ratio

3 ug / ml crystalline cellulose

Lactose 14.8 μg / ml

0.2 μg / ml magnesium stearate

According to a conventional capsule preparation method, the above ingredients were mixed and filled into gelatin capsules to prepare a pharmaceutical composition for improving obesity in a capsule form.

<Experiment 1> Confirmation of fat differentiation (producing) inhibitory effect of the isolate compound of Militaris cordyceps sinensis

The differentiation inhibitory effects of three different compounds, cordycepin, guanosine, and tryptophan, isolated from Militaris cordyceps, were measured.

1. Experimental Method

Adipocytes (3T3-L1) were differentiated by inducing differentiation-inducing substances IBMX, dexamethasone, cordycepin, guanosine, and tryptophan isolated from insulin and militaris cordyceps (0, 1). , 3, 10 ㎍ / ㎖) to determine the effect of inhibiting fat differentiation. That is, the fat cells stained with Oil Red O were lysed with isopropanol and absorbance was measured at 550 nm.

2. Experimental results

As a result, as shown in FIG. 6, it was confirmed that the fat accumulation was effectively reduced at 10 µg / ml when the three compounds cordycepin, guanosine, and tryptophan were administered alone.

<Experiment 2> Concomitant administration effect of the isolate compound of Militaris cordyceps sinensis

1. Experimental Method

The experiment was carried out under the same conditions as in Experimental Example 1.

2. Experimental results

As a result, as shown in Figure 7, the three compounds isolated from Militaris cordyceps, cordycepin, guanosine (guanosine), tryptophan (tryptophan) to reduce the fat accumulation when combined administration more effectively It was confirmed.

In other words, the existing anti-obesity effect of Militaris cordyceps was measured by only cordycepin as an indicator material, but the anti-obesity effect was largely varied according to the components of Militaris cordyceps. Cordycepin, guanosine, and tryptophan were selected as indicators of adipocyte differentiation of Militaris cordyceps.

Experimental Example 3 Content Analysis of Millitaris Cordyceps Sinensis by Extraction Solvent

In order to obtain a compound exhibiting the anti-obesity effect of Militaris Cordyceps sinensis, the extraction result was confirmed by varying the mixing ratio of ethanol and water solvent.

1. Experimental Method

After freeze-drying the Militaris cordyceps fruiting body and grinding it finely with a grinder, the mixture of ethanol and water (ethanol 100%, 75%, 50%, 0%), which is an extractant, was changed 10 times by volume. The extract was added and concentrated under reduced pressure at 40 ℃ and the content of the cordycepin (cordycepin), guanosine (guanosine), tryptophan (tryptophan) was measured for each extraction solvent.

2. Experimental results

As a result, as shown in Figure 8, cordycepin content was constant regardless of the ethanol solvent, guanosine content was the maximum content in 50% ethanol, tryptophan ethanol Inversely proportional to the content was confirmed.

Therefore, it is best to extract with 50% (50% water) ethanol solvent so that the content of cordycepin, guanosine, and tryptophan can be uniformly extracted. Confirmed.

Experimental Example 4 Cytotoxicity of the Separation Compounds of Militaris Cordyceps Sinensis

1. Experimental Method

Cytotoxicity of isolated compounds of Militaris cordyceps was confirmed by MTT assay method

In other words, cells were cultured by dispensing cells at 1 × 10 ⁴ / well in 96 well plates, followed by cordycepin, guanosine, and tritophan (differentiation of adipocyte differentiation of Militaris cordyceps). tryptophan) was incubated and reacted for 3 hours to dissolve the MTT solution was measured for absorbance at 570nm.

2. Experimental results

As a result, it was confirmed that the compounds of Militaris cordyceps sinensis as shown in Figure 9 does not show toxicity to adipocytes.

As described above, by providing a pharmaceutical composition for improving the harmless obesity of the human body having excellent anti-obesity effect and without the risk of side effects and toxicity, by providing the health functional foods with functional and safety to the people It can be seen that it can contribute to improving public health.

The present invention has been described with reference to preferred embodiments and experimental examples, and those skilled in the art to which the present invention pertains within the essential technical scope of the present invention and other embodiments of the detailed description of the present invention Examples and experimental examples may be implemented. The scope of the present invention is defined by the appended claims, and all differences within the scope of the claims are to be construed as being included in the present invention.

Claims (8)

Militaris Cordyceps derived cordycepin (cordycepin), guanosine (guanosine), comprising a compound consisting of two or more selected from tryptophan (tryptophan),
Pharmaceutical composition for improving obesity.
The method of claim 1,
The compound is characterized in that it is separated by extracting Militaris Cordyceps sinensis in a solvent and fractionated with butanol,
Pharmaceutical composition for improving obesity.
3. The method of claim 2,
Using a mixture of one or two selected from water or ethanol as the solvent,
Pharmaceutical composition for improving obesity.
The method of claim 3,
Made by mixing water and ethanol in a 1: 1 volume ratio as the solvent,
Pharmaceutical composition for improving obesity.
The method of claim 1,
The compound comprises a 50:50 weight ratio of two selected from cordycepin, guanosine, tryptophan
Pharmaceutical composition for improving obesity.
The method of claim 1,
The compound comprises a 45:55 weight ratio of two selected from cordycepin, guanosine, tryptophan
Pharmaceutical composition for improving obesity.
The method of claim 1,
The compound comprises cordycepin, guanosine, and tryptophan in a weight ratio of 20-30: 10-20: 50-60,
Pharmaceutical composition for improving obesity.
8. The method according to any one of claims 1 to 7,
The composition is characterized in that to inhibit the adipocyte differentiation,
Pharmaceutical composition for improving obesity.

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