KR102463297B1 - Novel use of extract of Artemisia annua - Google Patents

Abstract

The present invention provides treatment, improvement and/or treatment of at least one selected from bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease, and obesity of the wormwood extract or a pharmaceutical and/or food use for prophylaxis. According to the present invention, there is little or no possibility of side effects such as cancer, and one selected from bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease, and obesity. It has the advantage of being able to effectively treat, ameliorate and/or prevent abnormalities.

Description

Novel use of extract of Artemisia annua

The present invention relates to a novel use of wormwood extract, and more particularly, bone disease, menopausal disorder, cardiovascular disease, and neurodegenerative disease of the wormwood extract. Or it relates to the use of treatment, improvement or prevention of diseases or symptoms caused by estrogen deficiency, such as obesity.

Artemisia annua (Artemisia annua) is an annual herb belonging to the genus Asteraceae and medicinal herbs. It is traditionally used for medicinal purposes, and research results have shown that it has anticancer effects. Numerous pharmacological studies have been conducted, such as winning a Nobel Prize for development, but studies on specific associations with hormones have not been conducted.

On the other hand, estrogen (estrogen) is a hormone produced in the ovaries, it is known to have various functions in various organs in addition to the function of menstrual cycle control. For example, it regulates cholesterol production in the liver, maintains bone density in bones, and acts on the maturation of the uterine wall. In addition, it was recently found that it plays an important role in the survival of nerve cells and the process of adipogenesis.

When the concentration of estrogen in the body is decreased due to the stop of ovarian function in menopause, estrogen in the blood is decreased, and normal menopausal disorders including hot flushes, sweating, insomnia, depression, and headache appear. disease may occur. In addition, the risk of plaque formation in the vascular system increases due to the decrease in estrogen in the body, which leads to cardiovascular diseases such as atherosclerosis, and degenerative brain diseases such as Parkinson's and Alzheimer's due to nerve cell damage increase and lack of regulation of the adipogenesis process (Deroo BJ, Korach KS. (2006) "Estrogen receptors and human disease" J Clin Invest. Mar;116(3):561-70. PMID: 16511588).

Hormone therapy that administers animal-derived estrogen has been used to improve menopausal disorders caused by such estrogen reduction, but it is widely recognized that it has potential side effects such as breast cancer, cardiovascular disease, seizures, and blood clotting. reported in a randomized-placebo-controlled study (Rossouw JE, Anderson GL, Prentice RL, et al. (2002). "Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results From the Women's Health Initiative randomized controlled trial". JAMA 288 (3): 32133. PMID 12117397, Anderson GL, Limacher M, Assaf AR, et al. (2004). "Effects of conjugated equine estrogen in postmenopausal women with hysterectomy: the Women's Health Initiative randomized controlled trial" JAMA 291 (14): 170112. PMID 15082697).

Therefore, there is no or little possibility of such side effects, and it exhibits an estrogen effect, resulting in bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease, or obesity due to estrogen deficiency. It is necessary to develop a material having an improvement effect on (Obesity) and the like.

As a result of research efforts to discover substances for treating various diseases caused by estrogen deficiency, the present inventors have been able to improve diseases caused by lack of estrogen, , the present invention was completed by confirming that there is no side effect.

Accordingly, the present invention provides a bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease and obesity comprising an extract of Artemisia annua as an active ingredient. Obesity) for the purpose of providing one or more preventive or therapeutic pharmaceutical compositions selected from.

In addition, the present invention is selected from among bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease, and obesity comprising an extract of wormwood as an active ingredient. It is another object to provide a health functional food composition for one or more improvement or prevention.

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

In order to achieve the above object, the present invention provides a bone disease, menopausal disorder, cardiovascular disease, neurodegenerative comprising an extract of Artemisia annua as an active ingredient. It provides a pharmaceutical composition for preventing or treating at least one selected from disease) and obesity (Obesity).

In one embodiment of the present invention, at least one selected from bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease and obesity is estrogen deficiency may be due to

In another embodiment of the present invention, the bone disease may be one or more selected from osteoporosis, osteopenia or periodontal disease.

In another embodiment of the present invention, the cardiovascular disease may be atherosclerosis.

In another embodiment of the present invention, the degenerative brain disease may be at least one selected from Parkinson's disease or Alzheimer's disease.

In another embodiment of the present invention, the treatment or prevention may be due to the estrogenic activity of the wormwood extract.

In another embodiment of the present invention, the estrogenic activity may be selective estrogenic activity.

In another embodiment of the present invention, the selective estrogenic activity may be one that exhibits estrogenic activity, but does not selectively exhibit a cancer cell proliferation effect.

In another embodiment of the present invention, the cancer cells may be breast cancer cells or endometrial cancer cells.

In another embodiment of the present invention, the wormwood extract may be extracted with a solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.

In addition, the present invention is selected from among bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease, and obesity comprising an extract of wormwood as an active ingredient. It provides a health functional food composition for one or more improvement or prevention.

The wormwood extract of the present invention is a disease induced by estrogen deficiency: bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease, and obesity. It can effectively treat, ameliorate, or prevent at least one selected one or more, and does not have side effects such as carcinogenesis, so it can be widely used in the development of therapeutic agents for diseases caused by estrogen deficiency.

However, the effect of the present invention is not limited to the above effect, and it should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a result of the luciferase test whether Ishkawa cells were treated with an extract of the present invention and then estrogenic activity was confirmed.
Figure 2 is a result of the luciferase test whether MCF7 cells are treated with the extract of the present invention and then show estrogen activity.
3 is a result of luciferase test whether estrogen activity is shown after treatment of the extract of the present invention in MG63 cells.
Fig. 4 shows the side effects of the extract of the present invention by treating Ishkawa cells with the extract of the present invention, Fig. 4a shows the cell viability of the untreated control group and the positive control group treated with estradiol, and Fig. 4b is the ethanol-extracted dog poop. The cell viability of the mugwort extract, FIG. 4c is the result of confirming the cell viability of the wormwood extract extracted with hot water.
Figure 5 shows the side effects of the extract of the present invention by treating MCF7 cells with the extract of the present invention, Figure 5a shows the cell viability of the untreated control and the positive control treated with estradiol, Figure 5b is ethanol extracted The cell viability of the mugwort extract, FIG. 5c is the result of confirming the cell viability of the mugwort extract extracted with hot water.

As a result of research efforts to discover substances for treating various diseases caused by estrogen deficiency, the present inventors confirmed that the wormwood extract of the present invention can improve diseases caused by a lack of estrogen and has no side effects Thus, the present invention was completed.

Accordingly, the present invention provides a bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease and obesity comprising an extract of Artemisia annua as an active ingredient. Obesity) provides one or more prophylactic or therapeutic pharmaceutical compositions selected from.

In the present invention, the estrogen deficiency may be due to at least one selected from ovariectomy or menopause, the bone disease may be at least one selected from osteoporosis, osteopenia or periodontal disease, and the cardiovascular disease may be atherosclerosis, The degenerative brain disease may be at least one selected from Parkinson's disease or Alzheimer's disease, but is not limited thereto.

In the present invention, the treatment or prevention may be due to the estrogenic activity of the wormwood extract, and since the estrogenic activity of the wormwood extract of the present invention is a selective estrogenic activity, it means an estrogenic activity that does not have a cancer cell proliferation effect, which is a side effect, The cancer cells are not limited thereto, but may be breast cancer cells (MCF7) or endometrial cancer cells (Ishikawa).

In the present invention, estrogen activity means that it exhibits the same or similar activity as the physiological activity exhibited by estrogen produced in the body of mammals including humans.

Since the wormwood extract of the present invention has estrogenic activity, it can treat diseases caused by estrogen deficiency, but does not have the side effect of proliferating cancer cells, so it will be usefully used to treat diseases related to estrogen deficiency .

In the present invention, the extract can be extracted using a conventional solvent according to a conventional method known in the art for extracting an extract from a natural product, that is, under conditions of conventional temperature and pressure. For example, in the present invention, the extract may use one or more solvents selected from water, alcohols having 1 to 4 carbon atoms, and mixed solvents thereof. In addition, the method of extracting the extract may be extracted through various methods such as hot water extraction, cold extraction, reflux extraction, ultrasonic extraction, etc., and preferably, a hot water extraction method may be used, but is not limited thereto.

The prepared extract may then be filtered or concentrated or dried to remove the solvent, and both filtration, concentration and drying may be performed. For example, filtration may be performed using filter paper or a reduced pressure filter, concentration may be performed using a reduced pressure concentrator, drying may be performed using a freeze-drying method, and the like, but is not limited thereto.

The term "prevention" of the present invention refers to the administration of a pharmaceutical composition comprising the health extract provided in the present invention as an active ingredient to an individual who is expected to develop a disease induced by estrogen deficiency to inhibit or delay the onset of the disease. means all actions.

As used herein, the term "treatment" refers to any action that clinically intervenes to alter the natural process of an individual or cell to be treated, and may be performed during or to prevent clinical pathology. The desired therapeutic effect includes preventing the occurrence or recurrence of a disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, and alleviating the disease state. or temporary relief, remission and improvement of prognosis. For the purpose of the present invention, the treatment includes all actions of alleviating the disease induced by the estrogen deficiency by administering a pharmaceutical composition containing the extract of ragweed as an active ingredient to a patient who has developed a disease induced by estrogen deficiency. may be interpreted, but is not particularly limited thereto.

The present inventors confirmed the fact that the wormwood extract of the present invention can improve diseases caused by estrogen deficiency through specific examples.

In one embodiment of the present invention, it was confirmed through the luciferase experiment that the extracts of the present invention have estrogenic activity, thereby treating diseases caused by estrogen deficiency (see Example 2).

In another embodiment of the present invention, it was confirmed that the extract of the present invention has estrogenic activity, but does not induce cell proliferation such as cancer cells (see Example 3).

The pharmaceutical composition of the present invention may further include an appropriate carrier, excipient or diluent commonly used in the preparation of the pharmaceutical composition. The composition comprising a pharmaceutically acceptable carrier may be in various oral or parenteral formulations. In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used. Solid preparations for oral administration may include tablet pills, powders, granules, capsules, etc., and these solid preparations include one or more compounds and at least one excipient, for example, starch, calcium carbonate, sucrose or lactose. (lactose), gelatin, etc. can be mixed and prepared.

In addition to simple excipients, lubricants such as magnesium stearate, talc and the like may also be used. Liquid formulations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives are included. can Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

In addition, the pharmaceutical composition of the present invention is a group consisting of tablets, pills, powders, granules, capsules, suspensions, internal solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories It may have any one formulation selected from

On the other hand, as another aspect of the present invention, the present invention provides a method for preventing or treating a disease caused by estrogen deficiency, comprising administering to an individual a composition comprising an extract of wormwood as an active ingredient.

As used herein, the term “individual” refers to all animals, including humans, that are likely to develop a disease caused by estrogen deficiency or have developed it. By administering the composition of the present invention to an individual, it is possible to alleviate or treat diseases caused by estrogen deficiency.

As used herein, the term "alleviation" refers to any action in which a disease caused by estrogen deficiency is improved or beneficial by administration of the composition according to the present invention.

The composition of the present invention is administered in a pharmaceutically effective amount.

In the present invention, the term "administration" refers to introducing the pharmaceutical composition of the present invention to a subject by any suitable method, and the administration route may be administered through various routes, either oral or parenteral, as long as it can reach the target tissue.

The pharmaceutical composition of the present invention may be appropriately administered to a subject according to a conventional method, administration route, and dosage used in the art according to the purpose or necessity. Examples of administration routes include oral, parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal administration, and parenteral injection includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

In addition, an appropriate dosage and number of administration may be selected according to methods known in the art, and the amount and frequency of administration of the pharmaceutical composition of the present invention actually administered depends on the type of symptom to be treated, administration route, sex, and health condition. , diet, the age and weight of the individual, and the severity of the disease can be appropriately determined by various factors.

As used herein, the term "pharmaceutically effective amount" means an amount sufficient to inhibit or alleviate a disease caused by estrogen deficiency at a reasonable benefit/risk ratio applicable to medical use, and the effective dose level depends on the type and severity of the subject; Age, sex, drug activity, sensitivity to drug, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined.

The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. and may be administered single or multiple. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, and can be easily determined by those skilled in the art.

As another aspect of the present invention, the present invention includes bone disease, menopausal disorder, cardiovascular disease, neurodegenerative disease and It provides a health functional food composition for improving or preventing one or more selected from obesity (Obesity).

As used herein, the term “improvement” refers to any action that at least reduces a parameter related to the condition being treated, for example, the degree of pain symptoms.

In the health functional food composition of the present invention, the active ingredient may be added to food as it is or used together with other food or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of its use (for prevention or improvement). In general, in the production of food or beverage, the composition of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material. However, in the case of long-term ingestion for health and hygiene or health control, the amount may be less than or equal to the above range.

The health functional food composition of the present invention is not particularly limited in other ingredients other than containing the active ingredient as an essential ingredient in the indicated ratio, and may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. have. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate can be appropriately determined by a person skilled in the art.

In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and salts thereof, Alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like may be contained. These components may be used independently or in combination. The proportion of these additives may also be appropriately selected by those skilled in the art.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1. Preparation of wormwood extract

The mugwort extract of the present invention was extracted by adding an extraction solvent corresponding to 10 times the pulverized sample after pulverizing the mugwort wormwood, and the hot-water extract from wormwood was used by reflux extraction using distilled water, and ethanol from mugwort mugwort. (EtOH) extract was prepared by utilizing ultrasonic extraction (Ultrasonification extraction) using 70% ethanol. Reflux cooling extraction was performed twice for 1 hour under the condition of 100 °C heating mentle in a container containing the sample and distilled water, and then filtered under reduced pressure with filter paper (Whatman No. 2, Maidstone, England) and freeze-dried to powder got angry For ultrasonic extraction, the sample and 70% ethanol were put in a glass flask, mixed, and then placed in an ultrasonic water bath and extracted twice for 1 hour. After that, the obtained sample was filtered with a filter paper and then at a temperature of 45 ° C with a rotary vacuum concentrator. It was concentrated in vacuo and then pulverized.

Example 2. Confirmation of estrogenic activity of wormwood extract

Three types of cells known to be estrogen-reactive, Ishikawa (endometrial cancer cells), MCF7 (breast cancer cells) and MG673 (osteoblasts) were prepared, and then, to determine whether the extract of the present invention exhibits estrogenic activity, estrogen A luciferase assay was performed using an ERE-luc vector that exhibits luminescence when an estrogen-active substance binds to a receptor. Experiments were conducted with the control group (CON), the group treated with 10 ^-7 M estradiol (Estradiol, E2), the group treated with ethanol-extracted sagebrush extract (0.01, 0.1 μg/ml), and the group treated with hot water extract (0.01). , 0.1 μg/ml) was divided into groups treated with the experiment.

As a result, in Ishikawa cells, as shown in FIG. 1 , it was confirmed that the wormwood extract extracted with ethanol or hot water exhibited estrogen activity at a level similar to or higher than that of estradiol. As shown in Fig. 2 in MCF7 cells, it was confirmed that the wormwood extract extracted with ethanol or hot water showed a similar or higher level of estrogen activity to estradiol, and in MG63 cells, as shown in Fig. 3, the extract of the present invention This higher estrogen level was shown, and it was confirmed that the estrogen level was higher in the ethanol-extracted wormwood extract.

Example 3. Confirmation of side effects suppression effect by selective estrogenic activity of wormwood extract

Since it is known that if there is a side effect of an estrogen active ingredient, it can be a risk factor for cancer occurrence, so to check whether the extract of the present invention also has such side effects, Ishikawa (endometrium cancer cell) and MCF7 (breast cancer cell) ) for MTT assay was performed. The experiment was divided into an untreated control group, a positive control group treated with estradiol, and an experimental group treated with ethanol or hot water extract with concentrations of 0.00064, 0.00032, 0.016, 0.08, 0.4, and 2 μg/ml. Cell proliferation level of each group was performed in such a way as to confirm

As a result, in Ishikawa cells, it was confirmed that more cells were proliferated than the control group due to the side effect of estrogen activity in the positive control group as shown in FIG. 4a, but when the ethanol-extracted wormwood extract was treated (FIG. 4b) and hot water extracted gadfly When the mugwort extract was treated (FIG. 4c), it was confirmed that the cell proliferation was rather decreased as the concentration was increased, showing a low survival rate.

On the other hand, in MCF-7 cells, as shown in Fig. 5a, the positive control group was able to confirm that more cells were proliferated than the control group as a side effect of estrogen activity. In the case of treatment with the extracted wormwood extract (FIG. 5c), the cell proliferation rate was slightly higher than that of the control at a concentration of 0.00064 μg/ml, but when the extract of the present invention was treated by gradually increasing the concentration, the same as in the case of Ishikawa cells, breast cancer It was confirmed that the cell proliferation of the cell, MCF-7 cells, was reduced.

The inventors of the present invention were able to confirm that the extract of the present invention did not have side effects such as induction of cancer while exhibiting estrogenic activity through the above results.

The description of the present invention stated above is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. There will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (11)

It contains Artemisia annua extract as an active ingredient, exhibits estrogenic activity, exhibits a side effect suppression effect by selective estrogenic activity, and is for the prevention or treatment of menopausal disorders caused by estrogen deficiency, wherein the side effects are A pharmaceutical composition for cancer cell proliferation by an estrogen active ingredient.
delete delete delete delete delete delete delete The method of claim 1,
The cancer cell is a breast cancer cell or endometrial cancer cell pharmaceutical composition.
The method of claim 1,
The wormwood extract is a pharmaceutical composition, characterized in that extracted with a solvent selected from the group consisting of water, an alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof.
It contains wormwood extract as an active ingredient, exhibits estrogenic activity, exhibits a side effect suppression effect by selective estrogenic activity, and is for improvement or prevention of menopausal disorders caused by estrogen deficiency, and the side effects are estrogen active ingredients A health functional food composition that is cancer cell proliferation.

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