KR101770523B1 - Pharmacological compositions comprising Arteannuin B for treating or preventing of bone disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating osteoporosis comprising arteenine B as an active ingredient.
The pharmaceutical composition containing the artesanine B of the present invention has a low side effect and toxicity and exhibits a remarkable effect of inhibiting osteoclast formation than artemisinin, It is expected to be.

Description

[0001] The present invention relates to a pharmaceutical composition for preventing and treating bone diseases comprising arteenine B as an active ingredient,

The present invention relates to a composition for preventing and treating bone diseases, which comprises arteenine B as an active ingredient.

The osteoclast, which is a polynuclear large cell, has a function of destroying and absorbing bone tissue, and is said to be responsible for destroying the bone matrix and degrading the bone mineral. Activated osteoclasts have three or more nuclei, which require various hormones and factors to differentiate into osteoclasts from mature osteoclasts. Two of the most important factors are the macrophage colony stimulating factor (M-CSF) produced by osteoblasts and the receptor activator of nuclear factor-kappa B ligand (Mojtaba A., et al., Cancer Biol Ther., 7: 1,3-9; 1 (2008)). M-CSF is a cytokine that expresses osteoblasts and stromal cells, and plays an important role in the formation of osteoclasts and mainly plays an important role in cell proliferation, survival, and cytoskeletal formation (Kim et al. SY., Et al., J Korean Orthop Assoc., 44: 151-158 (2009)). Another important factor, RANKL, expresses osteoblast cells and attaches to RANK receptors in osteoclast precursor cells to induce osteoclast growth and differentiation (Mojtaba A., Cancer biology & Therapy, 7: 1,3-9 ; 1 (2008)). In addition, RANKL activates transcription factors such as c-fos, NFATc1 (nuclear Factor Of Activated T-Cells, cytoplasmic, calcineurin-dependent 1) and NF-kB to promote osteoclast differentiation, and PI-3K or ERK (LEE ZH., Et al., Biochem Biophys Res Commun., 305: 211-213 (2003)).

These osteoclasts cause abnormal bone tissue destruction and absorption due to imbalance with osteoblasts in the bone, resulting in osteoporosis in which bone mass and bone density are decreased, osteoporosis in bone, It is the cause of osteomalacia in which lime is removed, fibrous osteitis in which bone marrow is fibrous, periodontitis in which alveolar bone is lost, rheumatoid arthritis which causes joint destruction and deformation It is known. The imbalance between osteoclasts and osteoblasts can be caused by various causes such as bone metastasis of cancer cells, endocrine disorders, and hyperparathyroidism. Therefore, if it is possible to effectively inhibit the destruction and absorption of osteoclast by osteoclasts, various drugs and treatments for osteoclasts are being actively studied. Recently, a bisphosphonate-based therapeutic agent such as Fosamax (aldronate), Actonel (ingredient name: risedronate), Zometa (ingredient name: zoledronate), and the like have been used to treat bone damage caused by osteoclasts such as osteoporosis It is widely used. Most of these bisphosphonate preparations weaken the function of osteoclasts destroying bones and induce the death of osteoclasts to delay or suppress the loss of bones. Recently, however, there have been an increasing number of adverse events occurring in patients taking bisphosphonates, such as jawbone necrosis, severe atrial fibrillation, inability to bones or joints, and pain in the musculoskeletal system (Coleman RE, Br J Cancer, 98 : 1736-1740 (2008)). In addition, cancer cells that have metastasized to the bone in breast cancer and prostate cancer also promote the formation of osteoclasts, resulting in serious bone diseases.

Thus, there is a need to develop drugs that can complement the disadvantages of existing bisphosphonate preparations, have no toxicity, and can effectively inhibit bone resorption by osteoclasts.

On the other hand, Arteannuin B (B) is a major component contained in mugwort, which is biosynthesized through a different metabolic pathway than artemisinin, another major component of mugwort. Recently, artemisinin, which is one of the components of mugwort, has been known to be effective against cancer and osteoclast. However, little research has been done on Arte-anune-B.

Accordingly, the present invention relates to a pharmaceutical composition for preventing and treating osteoporosis, which comprises arteenine B as an active ingredient. The pharmaceutical composition of the present invention has few side effects and toxicity, and has a remarkable effect of inhibiting osteoclast formation than artemisinin Because it is visible, it is expected to be widely used in the medical field.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a pharmaceutical composition for preventing and treating osteoporosis, which comprises arteenine B as an active ingredient.

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

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions, and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and techniques of manufacture are not described in any detail, in order not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "embodiment" means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment "or" an embodiment "in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, a particular feature, form, composition, or characteristic may be combined in any suitable manner in one or more embodiments.

As used herein, "bone disease" means a disease caused by an osteoclast and osteoblastic imbalance in bone. Preferably osteoporosis, osteomalacia, fibrous osteitis, periodontitis, rheumatoid arthritis, metabolic bone disease, and the like caused by bone metastasis of cancer cells. But not limited to, diseases caused by bone resorption of osteoclasts.

In the present specification, " Artemisia annua " is an annual herbaceous plant of the dicotyledonous plant family. It is also called leaf mugwort, or dog mugwort, and is commonly found on roads, ponds and rivers. It is about 1m in height. It has no hairs on the whole grass and has a characteristic odor. The stem is green with many branches, the leaves are alternate, and the leaves are divided into 2 ~ 3 times finely. The leaves are 4 ~ 7 cm in length, and the leaves are small in shape and have fine hairs and glands on the outside. The center of the leaf is comb-shaped and the upper leaf is small. Flowers bloom in June-September with a greenish-yellow color. The head is round and 1.5 cm in diameter. The bushes are exposed. The involucre sculpture has no hairs and is arranged in 2 to 3 lines, and the outer sculpture is long oval and green. The fruit is about 0.7 mm in length.

Pharmacologically, in ancient China, it has been used to treat symptoms of infection with Plasmodium falciparum, such as malaria (Kooy and Sullivan, J Ethnophamacol., 150: 1-13 (2013)), It has been used for the treatment of cold, school, childhood, indigestion, dysentery.

In Korea standard KFDA standards, it can be used only for young leaves. Therefore, all currently marketed rings and essences are processed with only young leaves. However, it has been known that root, leaf and stems of Cryptomeria japonica are also effective as medicines.

In the present specification, " artemisinin " is a type of artichoke extract and is represented by the following formula (1).

Artemicinin is a sesquiterpene trioxane lactone compound which is separated from mugwort, and is known to exhibit an excellent therapeutic effect on malaria infection by removing malaria parasites through active oxygen production. Oral administration or intramuscular injection . Also toxoplasmosis (Toxoplasma gondii). It is also known to be effective against other infectious diseases such as Schistosoma, Pneumocystis carinii, Human Cytomegalovirus, Herpes simplex viruses, Hepatitis B, Hepatitis C and the like. In order to increase the efficacy of such artemisinin, studies have been made on the synthesis of dihydroartemisinin, Arteether, Artemether, Artesunate, and various derivatives thereof. Dihydroartemicinin has stronger anti-malarial efficacy than artemisinin and is known as the terminal metabolite of aether ether, atemeter and aethinate (Li QG., Et al., J Pharm Pharmacol., 50: 173-182 (1998)). In addition, recent studies have shown that artemicinin and dihydroartemisinin induce apoptosis in lung cancer, breast cancer, and prostate cancer, and inhibit invasiveness, leading to excellent anti-cancer effects.

In addition, artemisinin-glycolipid derivatives such as artemisinin-glycolipid have also been developed as derivatives of artemicinin. A representative artemisinin-glycolipid complex derivative is a complex derivative prepared by combining artemisinin with a glycolipid called daumone isolated from a beautiful nematode (Caenorhabditis elegans). It has been known that Daoumon is known as a substance that induces the nematode to the dormant and delays senescence and is known to inhibit angiogenesis of cancer (Jung M., et al., Eur J Med Chem., 44: 3120-3129 )). It has been reported that these artemisinin-glycolipid complex derivatives have higher anticancer effects in oral cancer, lung cancer and breast cancer than artemicinin and daumon (Ricci J., et al., Chem Pharm Bull., 59: 1471-1475 (2011)).

[Chemical Formula 1]

Artemisinin (AN) was firstly converted from precursor artemisinic aldehyde (AAA) to dihydroartemisinic aldehyde (DHAAA), and the dihydroartemisinic aldehyde was converted into dihydroartemic aldehyde After secondary conversion with dehydroartemisinic acid (DHAA), the dihydroartemisinic acid is converted to artemisinin through photooxidation and is biosynthesized.

In the present specification, " Arteannuin B " is a kind of artichoke extract and is represented by the following formula (2).

Artesanine B is transformed from artemisinic aldehyde (AAA), a precursor, to artemisinic acid (AA), and then biotransformed to artesanylin B through photooxidation.

Unlike artemisinin, which is an extract of the same dog mugwort, the research on the physiological function of arteenine B is very rare.

(2)

As used herein, the term " pharmaceutical composition " means a composition to be administered for a specific purpose. For the purpose of the present invention, the pharmaceutical composition of the present invention is a composition for the prevention and treatment of bone disease comprising arteenine B as an active ingredient, which may comprise a protein involved therein and a pharmaceutically acceptable carrier, excipient or diluent have. The above "pharmaceutically acceptable" carrier or excipient as referred to above means approved by the regulatory agency of the government or listed in government or other generally approved pharmacopoeias for use in vertebrates, and more particularly in humans do.

For parenteral administration, the pharmaceutical compositions of this invention may be in the form of a suspension, solution or emulsion in an oily or aqueous carrier, and may be prepared in the form of a solid or semi-solid. In addition, the pharmaceutical compositions of the present invention may include a formulating agent such as a suspending, stabilizing, solubilizing and / or dispersing agent and may be sterilized. The pharmaceutical composition may be stable under the conditions of manufacture and storage and may be preserved against the contaminating action of microorganisms such as bacteria or fungi. Alternatively, the pharmaceutical compositions of the invention may be in the form of a sterile powder for reconstitution with suitable carriers before use. The pharmaceutical compositions may be presented in unit-dose form, in micro needle patches, in ampoules, or in other unit-dose containers, or in multi-dose containers. Alternatively, the pharmaceutical composition may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid carrier, e. G., A vehicle immediately prior to use. Immediately, the injection solutions and suspensions may be prepared from sterile powders, granules or tablets.

In some non-limiting embodiments, the pharmaceutical compositions of the present invention may be formulated, or may be included in the form of microspheres in a liquid. In certain non-limiting embodiments, the pharmaceutical compositions of the present invention may include pharmaceutically acceptable compounds and / or mixtures thereof at a concentration of between 0.001 and 100,000 U / kg. Also, in certain non-limiting embodiments, the pharmaceutical compositions of the present invention may contain suitable excipients such as preserving agents, suspending agents, additional stabilizing agents, dyes, buffers, antimicrobial agents, antifungal agents, and isotonic agents such as, for example, . As used herein, the term "stabilizer" refers to a compound that is optionally used in the pharmaceutical compositions of the present invention to increase shelf life. In a non-limiting embodiment, the stabilizing agent may be a sugar, an amino acid, or a polymer. The pharmaceutical composition of the present invention may also include one or more pharmaceutically acceptable carriers, which may be solvents or dispersion media. Non-limiting examples of pharmaceutically acceptable carriers include water, saline, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycols), oils, and suitable mixtures thereof. Non-limiting examples of sterilization techniques applied to the pharmaceutical compositions of the present invention include filtration through a bacteria-inhibiting filter, sterilization of the terminals, incorporation of a sterile preparation, irradiation, sterilization gas irradiation, heating, vacuum drying and freeze drying do.

As used herein, the term " food composition " is used variously for improving and preventing bone diseases. The food composition containing the composition of the present invention as an active ingredient may be used in various foods such as beverages, gums, tea, vitamins Granules, tablets, capsules, confections, rice cakes, breads, and the like. Since the food composition of the present invention is modified and prepared from an existing food-grade food having little toxicity and side effects, it can be safely used for prolonged use for preventive purpose. When the composition of the present invention is contained in the food composition, the amount thereof may be added in a proportion of 0.1 to 100% of the total weight. Here, when the food composition is prepared in a beverage form, there are no particular limitations other than those containing the food composition at the indicated ratios and may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient. That is, natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, sucrose and the like, and sugar sugars such as polysaccharide, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol can do. Examples of the flavors include natural flavors (such as tau martin, stevia extract (for example, rebaudioside A and glycyrrhizin), and synthetic flavors (for example, saccharin and aspartame). The food composition of the present invention can be used as a food composition containing various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants, pectic acid and its salts, alginic acid and its salts, Stabilizers, antiseptics, glycerin, alcohols, carbonating agents used in carbonated beverages, etc. These components may be used independently or in combination. The ratio of such additives is usually in the range of 100 parts by weight per 100 parts by weight of the composition of the present invention But is not limited to, 0.1 to 100 parts by weight.

As used herein, " administration " means introducing the composition of the present invention to a patient by any appropriate method, and the administration route of the composition of the present invention may be administered through any conventional route so long as it can reach the target tissue have. Intravenous, intraperitoneal, intraperitoneal, intradermal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intradermal, intradermal, intraocular, Most preferably, the composition is provided as a broth of oral administration, but is not limited thereto.

The method of treatment of the present invention may comprise administering the pharmaceutical composition in a pharmaceutically effective amount. In the present invention, the effective amount may be determined depending on the kind of the disease, the severity of the disease, the kind and amount of the active ingredient and other ingredients contained in the composition, the type of the formulation and the age, body weight, general health condition, sex and diet, And the fraction of the composition, the duration of the treatment, the drug being co-administered, and the like.

In one embodiment of the present invention, there is provided a pharmaceutical composition for the prevention and treatment of bone diseases comprising arteenine B as an active ingredient, wherein the bone disease is osteoporosis, osteomalacia, fibrous osteitis ), Periodontitis, rheumatoid arthritis, and metabolic bone disease, wherein the bone disease is caused by bone resorption of osteoclasts. The present invention provides a pharmaceutical composition, wherein the bone disease is caused by bone metastasis of cancer cells.

In another embodiment of the present invention, there is provided a food composition for alleviating and alleviating bone diseases comprising arthaneun B as an active ingredient, wherein the bone disease is osteoporosis, osteomalacia, fibrous osteitis ), Periodontitis, rheumatoid arthritis and metabolic bone disease, characterized in that the bone disease is caused by osteoclastic bone resorption The present invention provides a food composition, wherein the bone disease is caused by bone metastasis of cancer cells.

Hereinafter, the present invention will be described in detail.

The present invention relates to a pharmaceutical composition for preventing and treating osteoporosis comprising arthaneunin B as an active ingredient. The pharmaceutical composition according to the present invention has few side effects and toxicity and can effectively prevent bone loss, It is expected to be utilized.

1 is a diagram showing a biosynthetic pathway and structure of artemisinic acid (AA), artemisinin (AN), or arteenine B (AB) according to an embodiment of the present invention.
FIG. 2 is a graph showing the inhibitory effect of artemisinic acid (AA), artemicinin (AN), or arteenine B (AB) on osteoclast formation according to an embodiment of the present invention.
Figure 3 shows the expression of c-Fos, NAFTc1 and GAPDH in mouse bone marrow macrophages treated with artemisinic acid (AA), artemicinin (AN), or arteenine B (AB), according to one embodiment of the present invention Fig.
FIG. 4 is a graph showing the results of converting the c-Fos expression level of FIG. 3 on the basis of GAPDH according to an embodiment of the present invention.
FIG. 5 is a graph showing a result of converting the expression amount of NAFTc1 in FIG. 3 on the basis of GAPDH, according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example  1: preparation of mouse bone marrow macrophages

For the preparation of mouse bone marrow macrophages, three-week-old male ICR mice (Nara Biotech, Korea) were disassembled from the cervical vertebrae, and the hindlimbs were removed using a saddle of the cervix. The hind limbs were cut off with surgical scissors, Immersed in α-MEM (minimum essential medium alpha; Gibco, USA). Then, the bones in the muscles were separated using a tweezers and transferred to a new α-MEM. The bone marrow cells were extracted by injecting 600 μl of α-MEM into the syringe and injecting it into the central spinal part of the separated leg bone 2 ~ 3 times . The extracted bone marrow cells were centrifuged to remove the supernatant, and fresh α-MEMs were mixed and bone marrow macrophages were isolated from the bone marrow cells using a histopaque (Sigma-Aldrich, USA) . (Gibco, USA), 10 volume% fetal bovine serum (FBS) (Gibco, USA), and M-CSF (macrophage-colony stimulating factor (R & D system Inc., USA), 30 ng / ml was added, and then the isolated mouse bone marrow macrophages were cultured.

Example  2: Arte Annuyin  B, Artemisicnik  mountain, Artemicinin  Experiment to confirm osteoclast formation inhibitory ability

In order to determine whether artesanine B, artemisinic acid, and artemisinin inhibit osteoclast formation induced by RANKL (receptor activator of nuclear factor kappa-B ligand), Park EK. et al., Biochem Biophys Res Commun., 325 (4): 1472-1480 (2004). And 40 or 80 μM of artemisinic acid, and 1 or 5 μM of arteenine B and artemisinin, respectively. After 5 x 10 ⁴ mouse bone marrow macrophages prepared according to the method of Example 1 were added to each well of a 96-well plate, α-MEM (1 volume% of antibiotic-antimicrobial solution , were cultured mouse bone marrow macrophages in 10 vol% of FBS, 30 ng / ㎖ of M-CSF, 100 ng / ㎖ of RANKL was added) each added at 200 ㎕ and 37 ℃, 5% CO ₂ conditions of cell culture medium . Then, the cells were exchanged for a new α-MEM culture containing each extract every other day for 5 days. Multicore osteoclasts were stained using a TRAP assay kit (Sigma-Aldrich, USA), and the number of osteoclasts with three or more nuclei was counted using an optical microscope. The results are shown in Table 1 and FIG.

Ingredients Component concentration (? M) RANKL-induced
Number of osteoclasts Osteoclast formation
% Inhibition - - 0 - RANKL 426 ± 5 0 Artemisinic acid RANKL + 40 399 ± 7 7 RANKL + 80 60 ± 8 86 Arte Annuyin B RANKL + 1 198 ± 12 54 RANKL + 5 8 ± 1 98 Artemisinin RANKL + 1 300 ± 11 30 RANKL + 5 53 ± 4 76

As shown in the above results, osteoclast formation was observed when 100 ng / ml of RANKL was treated. When artemicinic acid, arteenine B, or artemisinin were treated with RANKL, osteoclast formation It was confirmed that the concentration was decreased according to the concentration. In particular, arteenine B inhibited the differentiation of osteoclasts by 54% and 98%, respectively, at 1 or 5 μM concentration, which was more effective than that of artemisinin.

Example  3: Artemisinic acid , Arte Annuyin  B, Artemicinin  In osteoclasts, c-Fos, NFATc1  Expression inhibition confirmation experiment

When RANKL is treated on osteoclast precursor cells, RANKL binds to the RANK of osteoclast precursor cells and transmits signals into the cells. In the nucleus, the transcription factors c-Fos and NFATc1 are activated to differentiate osteoclasts (Asagiri M., et al., Bone, 40: 251-264 (2007)). Therefore, the present inventors investigated whether the osteoclast formation inhibitory effect of artemicinic acid, arteenine B, or artemicinin is related to these transcription factors. 10 ⁶ mouse bone marrow macrophages prepared by the method of Example 1 were cultured in a 60 mm ² culture dish with arteenine B (1 or 5 μM), artemicinic acid (40 or 80 μM), or artemisinin (1, Or 5 μM), and further cultured in medium containing RANKL (50 ng / ml) and M-CSF (30 ng / ml) for 48 hours and cells were obtained using a scraper. The obtained cells were added with protein lysis solution (150 mM NaCl, 0.5% Triton X-100, 50 mM Tris-HCl, pH 7.4, 20 mM EGTA, 1 mM DTT, 1 mM Na ₃ VO ₄ , protease inhibitor cocktail tablet ) Was added, and the mixture was allowed to stand on ice for 1 hour, followed by centrifugation (12,000 rpm, 10 min) to obtain a supernatant. The protein in the supernatant was quantitated by the BCA method (bicinchoninic acid method). Then, the supernatant was subjected to SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis), and the separated proteins were transferred to a PVDF membrane. The PVDP membrane was immersed in 5% skim milk solution dissolved in 0.1% TBST (Tris-buffered saline Tween 20) for 3 hours, and the primary antibody against c-Fos, NAFTc1 or GAPDH diluted 1: And the secondary antibody reacting specifically with the primary antibody was reacted. At the end of each step, non-specific antibody binding was stopped with 0.1% TBST, followed by ECL (Enhanced Chemiluminescence) solution, and then exposed to x-ray film to confirm changes in c-Fos, NAFTc1 and GAPDH. The results are shown in FIG. 3, and the results of converting the expression amounts of c-Fos or NAFTc1 on the basis of GAPDH are shown in FIGS. 4 and 5, respectively.

As a result, the expression of c-Fos and NFATc1 in RANKL-treated osteoclast precursor cells increased 2.8-fold and 1.6-fold, respectively, whereas RANKL and RANKL in osteoclast precursor cells treated with arteenine B (1 or 5 μM) Induced expression of c-Fos and NFATc1 was significantly inhibited. On the other hand, c-Fos expression was not significantly inhibited in artemisinin or artemisinic acid-treated osteoclasts with RANKL, and NFATc1 expression was treated with artemicinin 5 μM and artemisinic acid 80 μM , Respectively. As a result, at the same concentration, arteenuiin B inhibited the expression of c-Fos and NFATc1 by RANKL and was superior to artemicinin in inhibiting osteoclastogenesis.

Example  4: Culture of breast cancer cell line

Human basal-like / triple-negative breast cancer cell lines (MDA) and human basal-like / triple-negative breast cancer cell lines (MCF-7 and ZR-75-1) -MB-231, BT549, and HCC38) were purchased from the American Type Culture Collection (Manassas, Va., USA) and the Korean Cell Line Bank (Seoul, Korea). MCF-7 and MDA-MB-231 cells were cultured using 10% FBS-DMEM, and HCC38 and ZR-75-1 cells were cultured using 10% FBS-RPMI 1640. The BT549 cell line was cultured in RPMI 1640 medium containing 0.023 IU / ml insulin and 10% FBS. All breast cancer cells were cultured in a 5% CO ₂ incubator at 37 ° C.

Example  5: for breast cancer cell line Arte Annuyin  B, Artemisinic acid , Artemis Cytotoxicity of cinin

Artemisinin B, artemisinic acid, or artemicinin was treated to 80 [mu] M and cultured for 72 hours in a breast cancer cell line cultured by the method of Example 4. Then, a solution of MTT (3- (4,5-dimethythiazol-2-yl) -2,5-diphenyl tetrazolium bromide; Sigma, USA) was added to the solution at a concentration of 0.5 mg / , And then washed with PBS to completely remove the supernatant. Then, 200 μl of DMSO was added and reacted for 30 minutes, and absorbance was measured at 570 nm. Cell viability was calculated as a percentage of the absorbance of the experimental group (wells treated with arteenine B, artemisinic acid, or artemisinin) for the absorbance of the control (control group). The concentration (IC50) at which the cell viability was inhibited by 50% was calculated from the results and is shown in Table 2 below. As a result, it was found that the IC50 value of arteenine B was significantly lower than that of artemisinin and artemisinic acid.

Breast cancer cell line IC ₅₀ (mM) Artemisinin Artemisinic acid Arte Annuyin B MDA-MB-231 > 80 > 80 1.6 BT549 > 80 > 80 5.78 HCC38 > 80 > 80 13.4 MCF-7 > 80 > 80 25.0 ZR-75-1 > 80 > 80 20.0

From the above results, it was expected that arteenine B significantly inhibited the survival rate of breast cancer cells and inhibited the formation of osteoclasts, and thus could be useful for inhibiting bone damage by cancer cells.

Claims (8)

Pharmacology for the prevention and treatment of osteoporosis, osteomalacia, fibrous osteitis, periodontitis, rheumatoid arthritis, or metabolic bone disease, which comprise arteenine B as an active ingredient Composition.
delete The method according to claim 1,
Wherein the osteoporosis, osteomalacia, fibrous osteitis, periodontitis, rheumatoid arthritis, or metabolic bone disease is caused by bone resorption of osteoclasts. A pharmaceutical composition.
The method according to claim 1,
The pharmaceutical composition according to claim 1, wherein the osteoporosis, osteomalacia, fibrous osteitis, periodontitis, rheumatoid arthritis, or metabolic bone disease is caused by bone metastasis of cancer cells. Composition.
A food for improving and alleviating osteoporosis, osteomalacia, fibrous osteitis, periodontitis, rheumatoid arthritis, or metabolic bone disease, which contains arteenine B as an active ingredient Composition.
delete 6. The method of claim 5,
Wherein the osteoporosis, osteomalacia, fibrous osteitis, periodontitis, rheumatoid arthritis, or metabolic bone disease is caused by bone resorption of osteoclasts. Food composition.
6. The method of claim 5,
The method of claim 1, wherein the osteoporosis, osteomalacia, fibrous osteitis, periodontitis, rheumatoid arthritis, or metabolic bone disease is caused by bone metastasis of cancer cells. Composition.

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