WO2013032096A1 - Capric acid having inhibiting effect on osteoclast formation and pharmaceutical composition containing same - Google Patents

Abstract

The present invention provides an osteoclast formation-inhibiting agent containing capric acid as an active ingredient, and a pharmaceutical composition containing capric acid as an active ingredient for preventing or treating diseases due to bone loss. The capric acid, according to the present invention, inhibits the formation of an osteoclast by inhibiting the expression of MCP-1 and the expression of iNOS by means of STAT3(Ser⁷²⁷) activity in a RAW264.7 cell that is LPS-treated. Also, the capric acid according to the present invention reduces the expression of LPS-derived TRAP, which is an osteoclast differentiation marker. The pharmaceutical composition containing the capric acid, according to the present invention, is useful in treating various diseases that are induced from bone loss, wherein the diseases can be selected from a group consisting of osteoporosis, arthritis, and periodontitis. In addition, the pharmaceutical composition containing the capric acid, according to the present invention, is useful in preventing or treating asthma and renal failure.

Description

Capric acid having an inhibitory effect on osteoclast formation and pharmaceutical composition comprising the same

The present invention relates to capric acid having an inhibitory effect on osteoclast formation and a pharmaceutical composition comprising the same.

Osteoclasts are primarily derived from the myeloid hematopoietic mononuclear leukocyte / macrophage system. Osteoclasts are formed through various stages in the cell, including cell contact, binding, and differentiation. Osteoclasts are characterized by high expression of tatrate resistant acid phosphatase (TRAP), which can be used for the cytochemical differentiation of osteoclasts and their precursors. Increased osteoclast activity can lead to bone loss and resulting rheumatoid joints and osteoporosis. Thus, the regulation of osteoclastogenesis plays an important role in bone homeostasis.

Cell fusion plays an important role in regulating osteoclast formation. Nitric oxide (NO) and monocyte chemoattractant protein-1 (MCP-1) have been reported to enhance osteoclast formation by mediating cell fusion. Nitric oxide (NO) also increases the formation of osteoclasts by increasing actin remodeling in mononuclear pre-osteoclasts, thereby mediating fusion and the formation of multinuclear osteoclasts. Nitric oxide (NO) is a short-lived free radical that is involved in the regulation of many physiological processes such as vascular relaxation, neurotransmission, platelet aggregation, and immune responses. Moreover, it is formed by nitric oxide synthase (NOS) from oxygen and L-arginine. Three isoforms of the neuronal form (nNOS or NOS2), endothelial form (eNOS or NOS3) and inducible form (iNOS or NOS2) NOS have been identified. Of these three NOSs, iNOS is expressed in response to various inflammatory stimuli, which produce large amounts of nitric oxide (NO) by macrophages during inflammation. Receptor activator of nuclear factor-κB ligand (RANKL) and macrophage colony stimulating factor (M-CSF) are considered to be important for the formation of osteoclasts. Recent studies show that lipopolysaccharides (LPS) cause osteoclast differentiation and increase bone loss.

LPS, a cell wall product derived from bacteria, has long been recognized as a major factor promoting bone loss. LPS plays an important role in bone resorption, collecting inflammatory cells, synthesizing cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) and activating osteoclast formation and differentiation It is related to activities such as letting. LPS is a mediator that causes inflammation by osteoclasts through the nuclear factor-κB (NF-κB) pathway, three major mitogen-activated protein kinases (MAPKs), and extracellular signal-regulating kinases. Induces the production of 1/2 (extracellular signal regulated kinase 1/2, ERK1 / 2), c-Jun-N-terminal kinase (JNK) and p38.

Another important transcription factor associated with inducing cytokine induction is the signal transducer and activator of transcription (STAT). The STAT family is involved in the regulation of genes associated with the acute phase of inflammatory responses, cell growth and cell differentiation. The ability of STAT family proteins to homo- or hetero-dimerize is to alter gene transcription when they bind to specific response elements in the promoter of the target gene. In previous studies, the promoter region of the iNOS gene in murine macrophages revealed that it contained a STAT binding gamma activation site. STAT3 is associated with LPS-induced expression of iNOS and relies in part on Ser ⁷²⁷ phosphorylation, which is also required for translocation of the nucleus and DNA binding. In addition, activated STAT3 is involved in the regulation of cell growth, differentiation and survival and is essential for gp130-mediated osteoclast formation. Another STAT3 target gene, MCP-1 / Chemokine (CC motif) ligand 2 (Cc12), belongs to the CC chemokine family, which plays an important role in the recruitment and activation of white blood cells during polar inflammatory reactions. MCP-1 has been shown to play an important role in the development of atherosclerosis and other vascular diseases (such as blood vessels) by collecting mononuclear leukocytes into the arterial wall. Furthermore, MCP-1 is involved in cell-cell fusion of osteoclasts.

Meanwhile, capric acid, a medium chain fatty acid, is a major component of coconut oil and is known to improve the immune system by acting as a reinforcing agent, but the exact mechanism or effect thereof is not known.

Recently, there is increasing evidence that deficiency of certain fatty acids can lead to bone loss. In addition, some milk and fatty acids have been shown to inhibit osteoclast formation in bone marrow cultures and RAW264.7 cells. Fatty acids, only carboxylic acids with long hydrocarbon chains, are an important source of fuel for many tissues such as heart and skeletal muscle.

As a result of the above, it is important to prevent diseases related to various bone loss such as osteoporosis by inhibiting the differentiation of osteoclasts, and it is of interest to use capric acid as such therapeutic agent.

The inventors have confirmed that capric acid inhibits lipopolysaccharide-induced osteoclastogenesis in RAW264.7 cells, and completed the present invention based thereon.

Therefore, an object of the present invention is to provide an osteoclast formation inhibitor comprising capric acid as an active ingredient.

It is also an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of diseases caused by bone loss, including capric acid as an active ingredient.

It is also an object of the present invention to prevent or treat a disease caused by bone loss, comprising administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of capric acid as an active ingredient. It is to provide a way to.

It is also an object of the present invention to provide a pharmaceutical composition for preventing or treating asthma or kidney failure, which includes capric acid as an active ingredient.

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

The present invention provides an osteoclast formation inhibitor comprising capric acid as an active ingredient. In one embodiment of the invention, the capric acid inhibits LPS-induced osteoclast formation. In another embodiment of the invention, the capric acid inhibits MCP-1 expression. In another embodiment of the present invention, the capric acid inhibits iNOS expression, thereby blocking the production of nitric oxide (NO). In another embodiment of the present invention, the MCP-1 expression and iNOS expression is inhibited through the activity of STAT3 (Ser ⁷²⁷ ).

In addition, the present invention provides a pharmaceutical composition for preventing or treating a disease caused by bone loss, including capric acid as an active ingredient. In one embodiment of the invention, the disease is selected from the group consisting of osteoporosis, arthritis and periodontitis.

The present invention also provides a method of preventing or treating a disease caused by bone loss, comprising administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of capric acid as an active ingredient. To provide.

In addition, the present invention provides a pharmaceutical composition for treating or preventing asthma or kidney failure, including capric acid as an active ingredient.

Capric acid according to the present invention inhibits osteoclast formation by inhibiting MCP-1 expression and iNOS expression through the activity of STAT3 (Ser ⁷²⁷ ) in LPS-treated RAW264.7 cells. In addition, capric acid according to the present invention reduces LPS-induced TRAP expression, an osteoclast differentiation marker. The pharmaceutical composition comprising capric acid according to the present invention as an active ingredient is useful for the treatment of various diseases caused by bone loss, the disease may be selected from the group consisting of osteoporosis, arthritis and periodontitis. In addition, the pharmaceutical composition comprising capric acid according to the present invention as an active ingredient is useful for preventing or treating asthma or kidney failure.

1 shows morphological modification of RAW264.7 cells treated with LPS and capric acid.

Figure 2 is a diagram showing the effect of capric acid on LPS-induced osteoclast formation.

3 is a diagram showing the effect of capric acid on LPS-induced nitric oxide (NO) products.

Figure 4 shows that capric acid inhibits LPS-induced expression of iNOS and MCP-1 in RAW264.7 cells.

Figure 5 shows the effect of capric acid on LPS-induced phosphorylation of STAT3 in RAW264.7 cells.

Osteoclasts are multinucleated cells formed by cell fusion and are characterized by high expression of TRAP. One of the major factors of osteoclastogenesis induced by osteoblasts is the activation of receptor RANKL. Recently, it has been reported that LPS-induced osteoclast formation does not require RANKL and M-CSF. In addition, Hotokezaka et al. Said that RANKL-independent cell fusion of like osteoclasts can be induced by TNF-α, LPS and peptidoglycan. RAW264.7 cells act as progenitors for osteoclasts and differentiate into osteoclasts in response to LPS.

The inventors found that capric acid is the second medium-chain fatty acid, and that fatty acids are associated with bone density and reduce bone turnover, and furthermore, the capric acid is RAW264.7 It was confirmed that the inhibition of LPS-induced osteoclastogenesis in cells (lipopolysaccharide-induced osteoclastogenesis), and completed the present invention based on this.

Accordingly, the present invention provides an osteoclast formation inhibitor comprising capric acid as an active ingredient.

While LPS not only increases TRAP-positive multinucleated cell formation but also increases TRAP gene expression, capric acid can inhibit TRAP-positive multinuclear cell formation and TRAP gene expression (see FIG. 2). That is, capric acid according to the present invention can inhibit LPS-induced TRAP-positive osteoclast formation.

In addition, nitric oxide (NO), an important multifunctional signal molecule in bone, is produced by various cells in the basal or stimulation phase. Nitric oxide (NO) regulates bone formation, absorption, remodeling, mechanotransduction and treatment under physiological or pathological physiological conditions. Nitric oxide (NO) produced internally and supplied by nitric oxide (NO) donors are osteoclasts, osteoblasts, their precursors, and other cells in bone. They have a potent biphasic action that deeply affects their recruitment, proliferation, differentiation, activity and / or survival. iNOS is expressed in response to various inflammatory stimuli, which produce large amounts of nitric oxide (NO) by macrophages during inflammation. Nitric oxide (NO) is produced by iNOS and, more specifically, affects the formation and function of osteoclasts. Nitric oxide (NO) enhances osteoclast formation by mediating cell fusion. Based on cell type and stimulation, there are many binding sites for transcription factors in the iNOS region, including NF-ĸB, AP1, STATs and CCAAT / enhancer-binding proteins (C / EBP). Among the transcriptional regulators in the iNOS promoter region, NF-ĸB appears to be essential for cytokine products that cause LPS-induced inflammation. Abundant amounts of pre-inflammatory cytokines in the inflammatory site promote osteoclast differentiation and activity. Previous studies have shown that osteoclast formation is promoted by activating various intracellular signaling pathways, including MAPKs such as JNK, ERK, and P38, and transcription factors such as NF-κB, NFATcl and STAT.

However, it was confirmed that capric acid according to the present invention was ineffective on the NF-ĸB pathway, which means that capric acid is LPS-induced of pre-inflammatory cytokines IL-1β, TNF-α and IL-6. It is consistent with no effect on expression (see FIG. 5).

STAT, on the other hand, is another major signaling molecule involved in cytokine-induced inflammatory responses. Among the STAT families, STAT3 is an important transcription factor in the regulation of iNOS gene expression, and DNA binding of STAT3 has been shown to be affected by phosphorylation of Ser ⁷²⁷ and / or Tyr ⁷⁰⁵ . Previous studies have shown that the STAT-binding gamma active site (GAS) is essential for the expression of iNOS in LPS-induced RAW264.7 cells.

Low levels of nitric oxide (NO) may support osteoblast bone formation and osteoclast-mediated bone remodeling (both basic and cytokine-induced), while high oxidation Nitrogen (NO) levels and nitric oxide (NO) -generating compounds inhibit osteoclast formation and bone uptake and prevent bone loss in severely inflammatory or estrogen-deficient animals.

In one embodiment of the present invention, it was confirmed that capric acid according to the present invention has an inhibitory effect on nitric oxide (NO) production and iNOS mRNA expression (see FIGS. 3 and 4). In addition, capric acid according to the present invention also inhibited MCP-1 gene expression (STAT3 target gene).

Moreover, STAT3 mutant mice showed decreased bone density and bone volume and increased number of TRAP-positive osteoclasts. The results suggest that STAT3 plays a negative role in the regulation of osteoclastogenesis. In another embodiment of the present invention, it was found that, when treated with LPS alone, phosphorylation of STAT3 was significantly increased in Ser ⁷²⁷ , while phosphorylation was strongly reduced by capric acid (see FIG. 5B). ). In addition, LPS-induced iNOS and MCP-1 mRNA expression was inhibited by Stattic, a STAT3 inhibitor (see FIG. 5D). The results suggest that capric acid according to the present invention inhibits iNOS and MCP-1 expression through the activity of STAT3 (Ser ⁷²⁷ ).

The present invention also provides a pharmaceutical composition for preventing or treating a disease caused by bone loss, including capric acid as an active ingredient. The pharmaceutical composition according to the present invention can be usefully used for the treatment of diseases caused by bone loss, alleviation and prevention of symptoms, such diseases include, for example, osteoporosis, arthritis, periodontitis, but is not limited thereto. Does not.

Pharmaceutical dosage forms of the pharmaceutical compositions according to the invention can also be used in the form of their pharmaceutically acceptable salts, and can also be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The pharmaceutical composition of the present invention comprises a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may include physiological saline, polyethylene glycol, ethanol, vegetable oil and isopropyl myristate, and the like, but is not limited thereto.

The pharmaceutical compositions according to the invention may be formulated in ointments or creams for topical application and may be formulated in general saline, water-soluble solvents such as 5% dextrose or in ratios such as vegetable oils, synthetic fatty acid glycerides, higher fatty acid esters or propylene glycol. It may be formulated as an injection by dissolving, suspending or emulsifying in an aqueous solvent. Formulations of the present invention may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers and preservatives.

Preferred dosages of the pharmaceutical compositions according to the invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, preferably, the pharmaceutical composition according to the present invention is administered at 0.001 to 100 mg / kg body weight per day, more preferably at 0.01 to 30 mg / kg body weight. Administration may be administered once a day or may be divided several times.

The pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. The method of administration is not limited and may be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine epidural or cerebrovascular (intracerbroventricular) injection.

The present invention also provides a method of preventing or treating a disease caused by bone loss, comprising administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of capric acid as an active ingredient. To provide.

In the present invention, "individual" means a subject in need of treatment of a disease, and more specifically, human or non-human primate, mouse, rat, dog, cat, horse, cow, etc. Mean mammal. In addition, in the present invention, "pharmaceutical effective amount" may be adjusted in various ways depending on the weight, age, sex, health status, diet, time of administration, administration method, excretion rate and severity of the disease of the patient. It is obvious.

In addition, the present invention provides a pharmaceutical composition for preventing or treating asthma or kidney failure, including capric acid as an active ingredient. The pharmaceutical composition of the present invention has the effect of inhibiting nitric oxide (NO) and MCP-1, can be used as an asthma treatment, immunosuppressive agent, renal failure treatment and the like.

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.

EXAMPLE

Example 1. Cell Culture

RAW264.7, mouse macrophage cell line was obtained from ATCC. Cells were cultured in 10 mm plates and high glucose Dulbecco's Modified with 2 mM glutamine, antibiotics (1% penicillin / streptomycin) and 10% heat-inactive fetal bovine serum in a 37 ° C humidified incubator containing 5% CO _2. Incubated in Eagle Medium). LPS, capric acid, L-NMMA and TRAP kits were purchased from Sigma-Aldrich (St. Louis, MO, USA). P-JNK, JNK, p-ERK and ERK specific antibodies along with HRP-binding rabbit and mouse IgG antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies to IĸB-α were purchased from Santa Cruz (Santa Cruz, CA, USA). Nitric oxide (NO) detection kit was purchased from iNtRON Biotech (Seoul, Korea). RNeasy mini kit was purchased from Qiagen Inc (Valencia, CA, USA).

Example 2. Experimental Effect of Capric Acid on LPS-Induced RAW264.7 Cells

To determine the change in LPS induced morphology for capric acid, RAW264.7 cells were treated with various capric acid concentrations (0.1, 0.25 and 1 mM), and the results are shown in FIG. 1.

As shown in FIG. 1, the control group and the capric acid treated groups (0.1, 0.25 and 1 mM) showed no morphological changes, whereas the LPS-treated (1 μg / ml) group showed morphological changes. Showed. Compared to the LPS treatment group, the group treated with capric acid (1 mM) with LPS showed longer cell structures but no cell fusion. However, the group treated with capric acid at 1 mM concentration showed significant morphological changes.

Meanwhile, cell viability was measured using colorimetric MTT assay (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide assay). That is, RAW264.7 cells (5 × 10 ⁴ cells / well) were incubated in 96-well plates for 24, 48 and 72 hours and then treated with LPS with or without capric acid. After addition of 20 μl of MTT solution (5 mg / ml), cells were further incubated at 37 ° C. for 4 hours. The supernatant was then washed and insoluble formazan product was dissolved in DMSO. Then, the absorbance was measured using an ELISA reader at 570nm. Absorbance of formazan formed in untreated control cells was treated as 100% viability. MTT analysis showed that capric acid was not cytotoxic at the tested concentrations.

Example 3 Inhibition of TRAP-positive Multinuclear Cell Formation of Capric Acid

The effect of capric acid on osteoclast differentiation of LPS-induced RAW264.7 cells was measured using TRAP staining (Tartrate-resistant acid phosphate staining), an osteoclast marker, and the results are shown in FIG. 2. That is, RAW264.7 cells were incubated at a density of 5 × 10 ⁵ cells / well in 24-well plates for 12 hours and then treated with labeled compounds for an additional 24, 48 and 72 hours. The supernatant was then removed and the cells washed twice with PBS. Fixation solution was treated to cells and then washed with PBS. TRAP staining solution (50 mM acetate buffer, 30 mM sodium tartrate, 0.1 mg / ml Naphtol AS-MX phosphate, 0.1% w / v Triton X-100, and 0.3 mg / ml Fast Red Violet LB stain) for 1 hour Was treated. Thereafter, the staining solution was removed, the cells were washed twice with PBS, and the cell morphology was observed under a microscope.

As shown in FIG. 2A, LPS-induced osteoclast formation in RAW264.7 cells was confirmed for 1-3 days, although approximately 12% of RAW264.7 cells were TRAP-positive, multinucleated during the first 2 days after LPS stimulation. No production of cells was observed. Although there was still a small amount of TRAP-positive monocytes, on day 3 most of the TRAP-positive cells were confirmed to be multinucleated, and the results showed that the TRAP-positive cells were rapidly reduced by capric acid (1 mM). Suggest.

In addition, as shown in FIG. 2C, LPS-stimulated TRAP mRNA (osteoblast specific gene) expression was inhibited by capric acid.

Statistical analysis was performed using the Statistical Package for the Social Sciences (http://www.spss.com) v12.0 software. Each datum is expressed as mean ± S.E.M of different experiments under the same conditions. Statistical significance was compared between each treated group and control group by independent t-test. P-values less than 0.05 were considered statistically significant.

Example 4 Inhibition of LPS-Induced Nitric Oxide (NO) Formation of Capric Acid in RAW264.7 Cells

RAW264.7 cells were incubated at 96 × well plate for 12 hours at a density of 5 × 10 ⁴ cells / well and then treated with labeled compounds for 12, 18 and 24 hours. Supernatants were centrifuged from the cultured cells to remove cell debris and transferred to 96-well plates. The supernatant was then reacted using a Nitric oxide detection kit (iNtRON Biotech). The value was calculated by measuring the absorbance at 540 nm using a plate reader and the results are shown in FIG. 3. Statistical analysis was performed using the Statistical Package for the Social Sciences (http://www.spss.com) v 12.0 software. Each datum is presented as mean ± SEM of different experiments under the same conditions. Statistical significance was compared between each treated group and control group by independent t-test. P-values less than 0.05 were considered statistically significant.

As shown in FIG. 3, it was confirmed that capric acid significantly inhibited LPS-induced nitric oxide (NO) production over time. The results suggest that capric acid inhibits LPS-induced osteoclast formation by inhibiting nitric oxide (NO) production.

Example 5 LPS-Induced iNOS and MCP-1 Gene Upregulation Inhibition of Capric Acid in RAW264.7 Cells

RAW264.7 cells were treated with each compound (LPS 1 μg / ml, capric acid 1 mM) for 24, 48 and 72 hours and then washed with PBS. Total RNA was isolated by RNeasy mini kit (Qiagen), and total RNA concentration was measured using a spectrophotometer. Total RNA (1 μg) was converted to cDNA using cDNA Synthesis Master Mix (GenDEPOT; Barker, TX, USA). PCR was performed using Maxim PCR premix (iNtRON Biotech).

PCR primers are as follows:

TRAP-F; 5'-TCC CCT GGT ATG TGC TGG-3 'and

     R; 5'-GCA TTT TGG GCT GCT GA-3 '

INOS-F; 5'-GCA GAA TGT GAC CAT CAT CG-3 'and

     R; 5'-ACA ACC TTG GTG TTG AAG GC-3 '

MCP-1-F; 5'-GAA GGA ATG GGT CCA GAC AT-3 'and

      R; 5'-ACG GGT CAA CTT CAC ATT CA-3 '

GAPDH-F; 5'-CAT GAC CAC AGT CCA TGC CAT CAC T-3 'and

      R; 5'-TGA GGT CCA CCA CCC TGT TGC TGT A-3 '

The amplification sequence protocol was performed at 95 ° C. for 30 seconds, 58 ° C. for 30 seconds, and 72 ° C. for 45 seconds for each cycle. PCR products were separated by electrophoresis on 1% agarose gel and visualized by ethidium bromide staining, the results of which are shown in FIG. 4.

As shown in FIG. 4, when only LPS alone was used, iNOS and MCP-1 gene expression was significantly increased, whereas capric acid treatment significantly inhibited iNOS and MCP-1 expression. It was.

Example 6 Ser of Capric Acid in RAW264.7 Cells ⁷²⁷ LPS-Induced Phosphorylation Inhibition Experiment of STAT3

To clarify the molecular mechanism of capric acid during LPS-stimulated osteoclast formation, Western blot analysis measured the expression of phosphorylation of various signal molecules as well as transcription factors. RAW264.7 cells were lysed in protein extraction solution (iNtRON Biotech). Lysates were centrifuged at 4 ° C., 13,000 rpm for 20 minutes to remove cell debris. Protein concentration of the extract was measured by Bradford assay (Bio-Rad Laboratories; Hercules, CA, USA). Equal amounts of protein were separated by SDS polyacrylamide gel electrophoresis, and the separated proteins were transferred to nitrocellulose membranes and treated with 5% skim milk in TBST for 1 hour. The membrane was then reacted with various primary antibodies for 16 hours at 4 ° C. After washing, the membrane was reacted with HRP-goat anti-mouse IgG (H + L) binding antibody (Zymed Laboratories; San Francisco, CA, USA) diluted in TBST (1: 2,000) for 1 hour. After washing with TBST, blots were visualized with ECL agonists (Amersham Biosciences; Chalfont, Buckinghamshire, UK).

Treat cells with LPS for 0, 5, 15, 30, 60 and 120 minutes with or without capric acid to determine whether they activate ERK1 / 2, JNK, NF-κB, STAT1 and STAT3 The results thereof are shown in FIG. 5. β-actin was used as internal control.

As shown in Figures 5A and 5C, it was confirmed that capric acid had no effect on LPS-stimulated activation or expression of TNF-α, IL-1β and IL-6 of NF-κB, JNK and ERK1 / 2. .

On the other hand, as shown in Figure 5B, it was confirmed that LPS-induced phosphorylation of Ser ⁷²⁷ STAT3 is inhibited by capric acid.

In addition, Stattic, an inhibitor of STAT3, was used to determine whether LPS-induced expression of iNOS and MCP-1 was regulated by activation of STAT3, and as shown in FIG. 5D, LPS-induced iNOS and MCP It was confirmed that -1 gene expression was inhibited by Stattic. The results suggest that iNOS and MCP-1 expression is inhibited through the STAT3 pathway in RAW264.7 cells.

The foregoing description of the present invention is intended for illustration, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

The pharmaceutical composition comprising capric acid according to the present invention as an active ingredient is useful for the treatment of various diseases caused by bone loss, the disease may be selected from the group consisting of osteoporosis, arthritis and periodontitis. In addition, the pharmaceutical composition comprising capric acid according to the present invention as an active ingredient is useful for preventing or treating asthma or kidney failure.

Claims (9)

1. Osteoclast formation inhibitor comprising capric acid as an active ingredient.
2. The method of claim 1,

   The capric acid is characterized in that to inhibit the formation of lipopolysaccharide (LPS) -induced osteoclasts, osteoclast formation inhibitors.
3. The method of claim 1,

   The capric acid inhibits osteoclast formation, characterized in that it inhibits the expression of MCP-1 (monocyte chemoattractant protein-1).
4. The method of claim 1,

   The capric acid inhibits iNOS expression, thereby blocking the production of nitric oxide (NO), osteoclast formation inhibitor.
5. The method according to claim 3 or 4,

   The MCP-1 expression and iNOS expression is suppressed through the activity of signal transducer and activator of transcription 3 (STAT3) (Ser ⁷²⁷ ), osteoclast formation inhibitors.
6. A pharmaceutical composition for preventing or treating a disease caused by bone loss, including capric acid as an active ingredient.
7. The method of claim 6,

   The disease is selected from the group consisting of osteoporosis, arthritis and periodontitis, a pharmaceutical composition for preventing or treating a disease caused by bone loss.
8. A method of preventing or treating a disease caused by bone loss, comprising administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of capric acid as an active ingredient.
9. A pharmaceutical composition for preventing or treating asthma or kidney failure, including capric acid as an active ingredient.

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