WO2017123031A1 - Composition, for preventing, remedying or treating female postmenopausal osteoporosis, containing loganin or derivative of same as active ingredient - Google Patents

Abstract

The present invention relates to a composition, for preventing, remedying or treating female postmenopausal osteoporosis, containing loganin, a derivative of same or pharmaceutically acceptable salts of same as an active ingredient. Loganin, according to the present invention, is effective in inhibiting osteoclast differentiation as well as inhibiting postmenopausal bone density loss, bone microarchitecture degradation, a bone resorption marker increase in the blood and a decrease in bone formation markers. Therefore, loganin and a composition comprising same as an active ingredient, according to the present invention, are expected to be utilized as a pharmaceutical preparation for preventing, remedying and treating female postmenopausal osteoporosis.

Description

A composition for preventing, improving or treating postmenopausal osteoporosis in women, which contains loganan or a derivative thereof as an active ingredient.

The present invention relates to a composition for preventing, ameliorating or treating postmenopausal osteoporosis in women (type 1 osteoporosis, type 1) containing loganan, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

Estrogen, a representative hormone that symbolizes women, is an important female hormone that regulates the life of a woman from menstruation, pregnancy, and menopause. Estrogen is well known as a female hormone secreted mainly in the follicles, corpus luteum, and placenta in women's ovaries. Estrogens collectively include hormones such as estrone (E1), estradiol (E2), and estriol (E3). Estrogens affect a wide range of tissues and organs, especially the uterus, urinary system, breasts, skin, bones, and blood vessels, which are necessary to maintain flexibility and normality. Therefore, postmenopausal women may have various symptoms due to estrogen deficiency, and postmenopausal osteoporosis is one of the most serious symptoms (FIG. 1).

Osteoporosis refers to a condition in which bone mass in a unit volume is abnormally reduced compared to normal values according to gender, age, and race of a normal person. Even minor impacts such as bending back or sitting down can easily break bones, causing fractures in the hips, wrists, and vertebrae. It is divided into primary osteoporosis of postmenopausal osteoporosis (type 1 osteoporosis, type 1) and senile osteoporosis (type 2 osteoporosis, type 2) and secondary osteoporosis due to other causes, such as drugs (Fig. 2).

Postmenopausal osteoporosis (type 1 osteoporosis, type 1) occurs in women, after menopause, the lack of estrogen hormone occurs when the components of the bone is absorbed into the body tissues and calcium absorption through the intestine (Fig. 3). In particular, the rapid reduction of estrogen hormone promotes the differentiation and proliferation of osteoclasts, resulting in more bone resorption than bone formation, resulting in increased bone loss and reduced bone mass. Will occur (Figure 4). Depending on the person, it may progress rapidly after menopause.

The mechanism of action of postmenopausal osteoporosis is that when estrogen is reduced by menopause, interleukin-7 (IL-7) is greatly increased in bone marrow and thymus. This increase in IL-7 leads to an increase in the number of T-cells in peripheral blood, resulting in increased expression of tumor necrosis factor (TNF). In addition to TNF, an increase in cytokines such as RNAKL, M-CSF, IL-1, and IL-6 activates the differentiation and proliferation of osteoclasts. As a result, postmenopausal osteoporosis occurs due to a decrease in bone mass and bone density (FIG. 5).

Geriatric osteoporosis (type 2 osteoporosis, type 2) is caused by bone loss with increasing age in both men and women. Reduction of active vitamin D in the body results in less intestinal calcium absorption and decreased osteoblasts producing new bone cells.

Secondary osteoporosis is osteoporosis caused by various diseases or medications that affect the functions related to the production and maintenance of bone cells in the human body. Hyperthyroidism, parathyroidism, Cushing's syndrome, rheumatoid arthritis, hyperprolactinemia and the like are related, and steroid hormones, thyroid hormones, etc. may cause secondary osteoporosis.

At the onset of menopause, ovarian function decreases and ataxia reduces female hormones secreted from the ovary, which can lead to a variety of symptoms. Postmenopausal osteoporosis may occur due to a rapid decrease in bone density and a decrease in bone mass. Postmenopausal osteoporosis is a disease that greatly affects the quality of life because it can cause back pain or other diseases of the bone system and can easily fracture. In particular, early menopause or women who have had their ovaries removed before age 50 may be more vulnerable to postmenopausal osteoporosis.

The treatment for postmenopausal women with osteoporosis includes the combination-modified drugs such as Bonviva Plus (crude: bisphosphonate) and Aklasta (component: zoledronic acid 5mg injection). However, there is a side effect, the demand for natural products derived from therapeutics or health functional foods is increasing. Health functional foods such as calcium, vitamin D, and isoflavones, which are good for bone health, are used a lot, but the effect is limited. Accordingly, there is a need for the development of pharmaceutical preparations or dietary supplements for the prevention, improvement and treatment of more effective postmenopausal osteoporosis in women.

An object of the present invention is to provide a composition for preventing, improving or treating postmenopausal osteoporosis in women, which contains loganan, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient, and more specifically, after menopause. The present invention aims to provide a composition for preventing, improving and treating postmenopausal bone mineral density and osteoporosis caused by a decrease in female estrogen hormone.

The present invention provides a pharmaceutical composition for preventing or treating postmenopausal osteoporosis in women containing Loganin, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides a health food composition for preventing or improving postmenopausal osteoporosis in women, containing Loganin, a derivative thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention relates to a composition for preventing, ameliorating or treating postmenopausal osteoporosis in women, comprising loganan, a derivative thereof, or a pharmaceutically acceptable salt thereof as an active ingredient, wherein the loganan is an osteoclast It has good efficacy in inhibiting differentiation, and effectively inhibits postmenopausal bone mineral density reduction, bone microstructure density decrease, blood bone resorption marker increase and bone formation marker reduction. Accordingly, the present invention is expected to be useful as a pharmaceutical preparation for the prevention, improvement and treatment of postmenopausal osteoporosis in women with the composition comprising the lignin and the active ingredient.

Figure 1 shows the types and symptoms of female menopausal syndrome symptoms, postmenopausal osteoporosis is one of the most serious symptoms.

2 is a schematic diagram showing the mechanism of development of osteoporosis.

Figure 3 shows postmenopausal osteoporosis (type 1 osteoporosis, type 1) that occurs when the components of bone are absorbed into the body tissues as the estrogen hormone is insufficient after menopause and calcium absorption through the intestine decreases.

Figure 4 shows the pathogenesis of postmenopausal osteoporosis that occurs in post-menopausal estrogen deficiency.

5 shows the mechanism of action of postmenopausal osteoporosis.

Figure 6 shows the chemical structural formula of logannin.

Figure 7 is isolated from the mouse bone marrow (monocyte) that can be differentiated into osteoclasts (Fig. 7a), and treated with the concentration of 1 μg / ml, 5 μg / ml and 10 μg / ml of bone osteoclast The degree of differentiation into cells was analyzed by measuring TRAP (tartrate-resistant acid phosphatase) activity (FIG. 7B) and TRAP staining (FIG. 7C). Loganin significantly inhibited the differentiation of monocytes into osteoclasts. Indicates that there is an effect.

FIG. 8 was measured bone density change (FIG. 8A) during 12 weeks of administration of 10 mg / kg / day of loganine in a postmenopausal mouse model (ovary-extracted mouse). % Bone volume (BV) (FIG. 8C), trabecular thickness (Tb.Th) (FIG. 8D), trabecular number (Tb.N) (FIG. 8E), cavernous bone shochu The result of numerical analysis of the space (trabecular spacing, Tb.Sp) (FIG. 8F) shows that the effect of loganan significantly inhibiting the decrease in bone density and the density of bone microstructure by menopause.

9 is a serum metabolism marker RANKL (receptor activator of nuclear factor-kappaB ligand) (FIG. 9a) and OPG (osteoprotegerin) (OG) in the postmenopausal mouse model after 12 weeks of administration of 10 mg / kg / day Figure 9b) is a result of measuring the amount of protein in the blood and the ratio of OPG / RNAKL (Fig. 9c), the increase in bone resorption markers and decrease in bone formation markers caused by the post-menopause Significant inhibition is shown.

Accordingly, the present inventors have confirmed the in vitro and in vivo efficacy of a single compound, a compound that is effective in suppressing postmenopausal bone density and bone microstructure densities, which are manifested as decreased estrogen hormone in postmenopausal women, and the present invention. Was completed.

The present invention provides a pharmaceutical composition for preventing or treating postmenopausal osteoporosis in women containing Loganin, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

Specifically, the postmenopausal osteoporosis in women is due to a decrease in estrogen secretion, and the decrease in estrogen secretion decreases bone density, decreases bone microstructure densities, increases blood bone resorption markers and decreases bone formation markers. Can be derived.

More specifically, the bone resorption marker is a receptor activator of nuclear factor-kappa B ligand (RANKL), and the bone formation marker may be an osteoprotegerin (OPG), but is not limited thereto. .

Loganin of the present invention is represented by the formula (1), the molecular formula is C ₁₇ H ₂₆ O ₁₀ .

<Formula 1>

In the present invention, the pharmaceutically acceptable salt is an organic acid selected from the group consisting of oxalic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid and benzoic acid, or an inorganic acid selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid and hydrobromic acid. It may be an acid addition salt formed by.

When the composition of the present invention is a pharmaceutical composition, the pharmaceutical composition may be formulated as a cream, gel, patch, spray, ointment, warning, lotion, linen, pasta and cataplasma. On the other hand, the pharmaceutical composition may include a pharmaceutically acceptable carrier in addition to the Loganin, such a pharmaceutically acceptable carrier is commonly used in pharmaceutical formulations, lactose, dextrose, Sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl Hydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto. In addition, the pharmaceutical composition may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like as an additive.

The pharmaceutical composition is determined according to the degree of symptoms of postmenopausal osteoporosis in women, a topical administration is usually preferred. In addition, the dosage of the active ingredient in the pharmaceutical composition may vary depending on the route of administration, the degree of the disease, the age, sex, and weight of the patient, and may be administered once to several times daily.

The present invention also provides a health food composition for preventing or improving postmenopausal osteoporosis in women, containing Loganin, a derivative thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The health food composition may be provided in the form of a powder, granules, tablets, capsules, syrups, beverages or pills, wherein the health food composition is in addition to other foods or food additives other than loganan (Loganin) according to the present invention as an active ingredient It can be used together and appropriately used according to a conventional method. The mixed amount of the active ingredient can be suitably determined depending on the purpose of use thereof, for example, prophylactic, health or therapeutic treatment.

An effective dose of Loganin contained in the health food composition may be used in accordance with the effective dose of the pharmaceutical composition, but in the case of long term intake for health and hygiene purposes or for health control purposes, It may be less than the above range, it is clear that the active ingredient can be used in an amount above the above range because there is no problem in terms of safety.

There is no particular limitation on the kind of the health food, for example, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinks, tea, Drinks, alcoholic drinks, vitamin complexes, etc. are mentioned.

Hereinafter, examples will be described in detail to help understand the present invention. However, the following examples are merely to illustrate the content of the present invention is not limited to the scope of the present invention. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example 1 Chemical Structure of Loganin

The test single compound used in the animal experiments in the present invention is Loganin. Loganin is an iridoid glycoside of the name Derived from the Loganiaceae, the chemical formula is C ₁₇ H ₂₆ O ₁₀ (Fig. 6). Loganin is a naturally occurring single compound that is found in horses (nux vomica) and cornus officinalis.

Example 2 Inhibitory Effect of Loganin on Osteoclast Differentiation

Osteoclasts are derived from hematopoietic stem cells and are responsible for bone resorption that destroys aged bone.Bone formation by osteoblasts and bone resorption by osteoclasts Bone remodeling is achieved through a balanced action of resorption. After menopause, in which female estrogens were sharply reduced, we investigated the efficacy of loganan on inhibition of osteoclast differentiation, especially because bone resorption is increased due to the differentiation and proliferation of osteoclasts.

For the differentiation activity of osteoclasts, primary monocytes, which are the source cells of osteoclasts, were isolated and cultured from 6-week-old mouse bone marrow. After treatment with CD11b, a mononuclear cell positive marker antibody, mononuclear cells were properly isolated and cultured from mouse bone marrow using a fluorescence activated cell sorter (FACS) (FIG. 7A). Mouse mononuclear cells were treated with osteoclasts for M-CSF (30 ng / ml) and LANKL (50 ng / ml), which induced differentiation into osteoclasts, and induced differentiation into osteoclasts for 3 days. In addition, when the osteoclast differentiation was induced, physiological saline was added to the negative control group and rogannin was added to the medium at concentrations of 1 μg / ml, 5 μg / ml, and 10 μg / ml. The degree of differentiation activity of monocytes into osteoclasts was analyzed by measuring and staining of TRAP (Tartrate-resistant acid phosphatase) activity. After 3 days of cell culture, the cells were washed with saline, and then TRAP activity was analyzed by absorbance measurement at a wavelength of 405 nm and by microscopic observation after TRAP staining. Compared to the control group treated with physiological saline, statistically in all experimental groups treated with the concentration of loganan at 1 μg / ml (Lo1), 5 μg / ml (Lo5) and 10 μg / ml (Lo10) Significantly the differentiation of monocytes into osteoclasts was inhibited (FIG. 7B) (*: p <0.05 vs. negative control). In addition, the number of osteoclasts differentiated in the experimental group (Induction + Loganin) treated with osteoclast induction and roganine (10 μg / ml) was significantly reduced compared to the osteoclast induction control (Induction) in the TRAP staining experiment. 7c). In other words, it was found that loganine has an excellent effect on inhibiting osteoclast differentiation.

< Example  3> Ovariectomy in a Postmenopausal Mouse Model Loganine  Inhibition of post-menopausal osteoporosis

To evaluate the in vivo efficacy of logannin, a 10-week-old ovaryctomized-mouse (OVX mouse) mouse was used as a postmenopausal animal model (additional breeding for 2 weeks to recover from surgery after 8-week-old ovarian resection). . As a control group, Shame mice group (open control group) without ovarian resection, OVX mouse group (negative control group) administered physiological saline only to ovarian resected mice, and strontium chloride, a compound having bone mineral density improvement effect in ovarian resected mice ( SrCl ₂₎ oral administration as a 10mg / kg / day dose for (oral injection) a SrCl ₂ The mouse group was used as a positive control in the bone density improvement efficacy experiment. The experimental group was orally injected with loganine at a 10 mg / kg / day dose. 10-week-old sham-operated and ovarian abdominal ddY female mice were purchased from Central Experimental Animal Co., Ltd. and moved to a clean animal breeding area after 1 week of acclimation in the Quarantine Office of Ajou University. The weight of each mouse was measured, and group separation was performed so that there was no statistically significant weight difference between the experimental groups. Loganine to be used in the experiment was made of a test solution in an aqueous solution, and the sterilization operation was performed by gamma irradiation to the Soya Green Tech Co., Ltd., a radiation irradiation company for the import of experimental animal center.

Initial bone mineral density (BMD) was measured with a PIXImus bone densitometer at the start of animal testing. Mice were anesthetized by injecting 50 ul of a mixed anesthetic of zoletil and rompun (diluted 1: 2 mixture with physiological saline in a 2: 3 ratio), and then fixed in a bone density measuring frame and measuring bone density. After 6 weeks and 12 weeks after the administration of Loganin to the mouse, the bone density of the mouse was measured with a PIXImus bone densitometer, and after 12 weeks of experiments, blood collection and femoral bones were taken and micro-CT was taken. bone volume (BV), trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular spacing (Tb.Sp) were numerically analyzed.

Significant decrease in bone density increase after 6 and 12 weeks of bone in ovarian-depleted OVX mice compared to normal Shame mice without ovarian removal (FIG. 8A), and bone microstructure densities on micro-CT images A marked drop in was shown (FIG. 8B). In addition, the bone volume ratio (Fig. 8c), the thickness of the spongy bone shochu (Fig. 8d), the number of spongy bone shochu (Fig. 8e) was significantly low and the spongy bone shochu space (Fig. 8f) was high. However, in the experimental group administered Loganin at a dose of 10 mg / kg / day for 12 weeks, the decrease in bone density caused by menopause and the decrease in the density of bone microstructures were suppressed (FIGS. 8A and 8B), and the volume fraction of bone and sponge sponge The decrease in thickness, the number of spongy bone marrow and the increase in the space of bone marrow were also suppressed (Figs. 8C, 8D, 8E, 8F). These results indicate that the positive control SrCl ₂ Similar to the results of mice. In addition, the statistical significance of the efficacy was confirmed (*: p <0.05 vs. OVX negative control).

Next, changes in blood metabolic markers were examined. Twelve weeks after the postmenopausal mouse model experiment, mouse blood was collected and serum was isolated to determine the receptor activator of nuclear factor-kappa B ligand (RANKL), a bone resorption marker, and the osteoprotegerin (OPG), a bone formation marker. Blood protein levels were analyzed by ELISA method. Compared to normal Shame mice without ovarian removal, OVX mice without ovary showed an increase in RANKL (FIG. 9A), a decrease in OPG (FIG. 9B), and a decrease in OPG / RANKL ratio (FIG. 9C). However, in the experimental group administered Loganin at a dose of 10 mg / kg / day for 12 weeks, an increase in the blood bone resorption marker RANKL, a decrease in the bone formation marker OPG, and a decrease in the OPG / RANKL ratio were all inhibited (Fig. 9a, 9b, 9c). These results indicate that the positive control SrCl ₂ Similar to the results of mice. In addition, the statistical significance of the efficacy was confirmed (*: p <0.05 vs. OVX negative control).

Claims (7)

1. A pharmaceutical composition for preventing or treating postmenopausal osteoporosis in women, which contains loganan, a derivative thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
2. The pharmaceutical composition for preventing or treating postmenopausal osteoporosis in women according to claim 1, wherein the postmenopausal osteoporosis in women is caused by a decrease in estrogen secretion.
3. The postmenopausal female postmenopausal osteoporosis according to claim 2, wherein the decreased estrogen secretion induces a decrease in bone density, a decrease in bone microstructure density, an increase in blood bone resorption markers and a decrease in bone formation markers. Prophylactic or therapeutic pharmaceutical composition.
4. The pharmaceutical composition for preventing or treating postmenopausal female osteoporosis according to claim 3, wherein the bone resorption marker is a receptor activator of nuclear factor-kappa B ligand (RANKL).
5. The pharmaceutical composition for preventing or treating osteoporosis in postmenopausal women according to claim 3, wherein the bone formation marker is OPG (osteoprotegerin).
6. Health food composition for preventing or improving postmenopausal osteoporosis in women containing logann (Loganin), derivatives thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
7. The method of claim 5, wherein the health food is a post-menopausal osteoporosis prevention or improvement health food composition for female, characterized in that it has any one form selected from the group consisting of tablets, capsules, powders, granules, liquid and pill.

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