JP2024000101A - Osteoclast differentiation inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a safe agent for osteoporosis.

SOLUTION: The foregoing problem can be solved by an osteoclast differentiation inhibitor that comprises ground or extracted substances from plants selected from the group consisting of coriander, bay laurel, cardamon, chili pepper, pepper, sage, and clove.

SELECTED DRAWING: None

COPYRIGHT: (C)2024,JPO&INPIT

Description

The present invention relates to an osteoclast differentiation inhibitor. According to the present invention, osteoporosis can be prevented or treated.

Osteoporosis is common in women and mainly occurs due to hormonal imbalance after menopause. It is also said to be caused by aging, genetic predisposition, unbalanced diet, extreme dieting, smoking, or excessive drinking.

Japanese Patent Application Publication No. 2016-104759

Bisphosphonate-based drugs are known as drugs for treating osteoporosis (Patent Document 1). However, there have been reports of side effects, and there are hopes for the development of safe osteoporosis drugs.
The purpose of the present invention is to provide a safe osteoporosis drug.

As a result of intensive research into safe osteoporosis drugs, the present inventor surprisingly discovered that a crushed product or extract of a specific herb inhibits osteoclast differentiation.
The present invention is based on these findings.
Therefore, the present invention
[1] An osteoclast differentiation inhibitor comprising a crushed product or extract of a plant selected from the group consisting of coriander, bay, cardamom, chili pepper, pepper, sage, and clove;
[2] The osteoclast differentiation inhibitor according to [1], wherein the extract is an extract using a polar organic solvent, an aqueous solvent, or a mixed solvent containing a polar organic solvent or an aqueous solvent;
[3] The plant is coriander fruit, bay leaf, cardamom fruit, chili pepper fruit, pepper fruit, sage leaf, stem, or flower spike, or clove flower bud, [1] or [2] The osteoclast differentiation inhibitor described in
[4] A pharmaceutical composition for preventing or treating osteoporosis, comprising the osteoclast differentiation inhibitor according to any one of [1] to [3], and [5] A pharmaceutical composition according to any one of [1] to [3]. A food composition for preventing or treating osteoporosis, comprising an osteoclast differentiation inhibitor,
Regarding.

According to the osteoclast differentiation inhibitor of the present invention, osteoclast differentiation can be suppressed.

It is a graph showing the inhibitory effect of coriander extract on osteoclast differentiation. It is a graph showing the inhibitory effect of bay extract on osteoclast differentiation. It is a graph showing the inhibitory effect of chili pepper extract on osteoclast differentiation. It is a graph showing the inhibitory effect of pepper extract on osteoclast differentiation. It is a graph showing the inhibitory effect of clove extract on osteoclast differentiation. It is a graph showing the inhibitory effect of cardamom extract on osteoclast differentiation. It is a graph showing the inhibitory effect of sage extract on osteoclast differentiation.

The osteoclast differentiation inhibitor of the present invention includes a crushed product or extract of a plant selected from the group consisting of coriander, bay, cardamom, chili pepper, pepper, sage, and clove.

"coriander"
Coriander is a member of the Apiaceae family and is used as a spice. As a spice, the whole plant is used, the fruit and seeds are used as coriander seeds, and the leaves and stems are used as coriander leaves or coriander. In the present invention, the part to be used of coriander is not particularly limited, and may include the whole plant, root, stem, leaf, flower, fruit, or seed, or a mixture of at least two or more thereof. Preferably it is fruit. Coriander may be used in its raw state or dried (for example, freeze-dried) when performing a crushing operation or an extraction operation. In the case of extraction, it is preferable to process it into a crushed product or powder state so that the extraction efficiency is improved.

"laurel"
Laurel is a plant of the Lauraceae family, genus Laurel, and the leaves of the bay tree are called laurel. It is said to be native to the Mediterranean coast, and has a refreshing, distinct aroma, and is used for scenting. It eliminates the smell of meat and is used as a spice in European-style curry and stew dishes such as pot-au-feu. Laurel may be used fresh or dried (for example, freeze-dried) when performing the crushing or extraction operation. In the case of extraction, it is preferable to process it into a crushed product or powder state so that the extraction efficiency is improved.

"cardamom"
There are two types of cardamom: green cardamom and black cardamom. The cardamom used in the present invention is green cardamom (Elettaria cardamomum). Green cardamom is a perennial herb of the Zingiberaceae family that is native to India, Sri Lanka, and the Malay Peninsula, and is also called Shozuku. The part where cardamom is used is not particularly limited, and may include the whole plant, root, stem, leaf, flower, fruit, or seed, or a mixture of at least two or more thereof, but preferably the fruit. be. The green cardamom fruit is dried, including the seeds, and used as a spice, either as is or ground into a powder. In the present invention, cardamom may be used raw or dried (for example, freeze-dried) when performing the pulverization or extraction operation. In the case of extraction, it is preferable to process it into a crushed product or powder state so that the extraction efficiency is improved.

《Chili pepper》
Chili pepper is a plant of the Capsicum genus of the Solanaceae family, native to Central and South America, and the fruit is used as a spice. There are various types of chili peppers, and the chili peppers used in the present invention are not limited, but for example, red chili peppers or green chili peppers can be used.
The part of chili pepper to be used is not particularly limited, and may include the whole plant, roots, stems, leaves, flowers, fruits, or seeds, or a mixture of at least two of these, but preferably the fruit. be. Chili peppers may be used fresh or dried (for example, freeze-dried) when performing the crushing or extraction operation. In the case of extraction, it is preferable to process it into a crushed product or powder state so that the extraction efficiency is improved.

"pepper"
Pepper is a climbing plant belonging to the Piperaceae family, and the fruit is used as a spice. In the present invention, the part used for pepper is not particularly limited, and may include the whole plant, root, stem, leaf, flower, fruit, or seed, or a mixture of at least two or more thereof, Preferably it is fruit. Pepper may be used fresh or dried (for example, freeze-dried) when performing the pulverization or extraction operation. In the case of extraction, it is preferable to process it into a crushed product or powder state so that the extraction efficiency is improved.

《Sage》
Sage is a perennial or evergreen shrub of the Lamiaceae family that is native to the Mediterranean Sea. The height is about 50-70 cm, and purple or white lip-like flowers bloom from May to July. The oblong, stalked leaves are opposite and have fine curls on the surface, and the leaves are used as a spice.
The parts of sage to be used are not particularly limited, and may include the whole plant, roots, stems, leaves, flowers, fruits, or seeds, or a mixture of at least two or more thereof, but preferably leaves, It may be a stem or a flower spike, or a combination of two or more of these (leaves and stems, leaves and flower spikes, stems and flower spikes, or leaves, stems, and flower spikes). Sage may be used raw or dried (for example, freeze-dried) when performing the crushing or extraction operation. In the case of extraction, it is preferable to process it into a crushed product or powder state so that the extraction efficiency is improved.

"cloves"
Clove is a tree belonging to the Myrtaceae family, and the flower buds are used as a spice. In the present invention, the parts used for pepper are not particularly limited, and examples include the whole plant, roots, stems, leaves, flowers, fruits, flower buds, or seeds, or a mixture of at least two or more thereof. However, flower buds are preferred. Cloves may be used fresh or dried (for example, freeze-dried) during the crushing or extraction operation. In the case of extraction, it is preferable to process it into a crushed product or powder state so that the extraction efficiency is improved.

《Crushed material》
The pulverized plant material in the present invention is not particularly limited as long as the plant is in a pulverized state. It is. Further, powders formed or granulated into cubes, blocks, or granules can also be preferably used. Processing for processing into pulverized products is not particularly limited, but for example, plants can be processed by any method commonly used by those skilled in the art, using equipment or tools for pulverization such as crushers, mills, blenders, mixers, and stone mills. One example is the process of crushing the body. The plant body may be dried before pulverization. Examples of drying methods include freeze drying, reduced pressure drying, blow drying, and heat drying.

When the plant contained in the osteoclast differentiation inhibitor of the present invention is a crushed plant, the average longest diameter of the crushed plant is, for example, 0.01 to 2 mm, preferably 0.01 to 2 mm, although it is not limited to this. 0.01 to 1.5 mm, more preferably 0.01 to 1 mm, still more preferably 0.01 to 0.75 mm, and most preferably 0.01 to 0.5 mm. In addition, 90% by weight or more of the crushed plant material is 0.01 to 2 mm, preferably 0.01 to 1.5 mm, more preferably 0.01 to 1 mm, still more preferably 0.01 to 0.75 mm, and most preferably Preferably, those having a longest diameter of 0.01 to 0.5 mm can be used. In addition, 90% by weight or more of the crushed plant material is 8.6 mesh (2 mm), 10 mesh (1.7 mm), 16 mesh (1 mm), or 30 mesh (0.5 mm) in the JIS test sieve mesh conversion table. You can use anything that passes through. The average longest diameter of a plant can be measured using a known device for measuring particle size. Alternatively, it can also be calculated by arbitrarily selecting 100 crushed plants, measuring their longest diameters using a stereomicroscope, and calculating their average.

《Extract》
Extracts containing active ingredients can be extracted by conventional extraction methods used for extracting ingredients derived from plants. Extraction methods include, but are not limited to, solvent extraction, steam distillation, expression (direct, hot, or cold), or supercritical extraction. Further, a combination of these extraction methods may be used, for example, a method of squeezing followed by solvent extraction, but solvent extraction is preferable. The plants used for extraction may be used fresh or dried. Further, in order to improve the extraction efficiency, the extract may be extracted after being processed into a crushed product or powder state.

(solvent extraction method)
The extraction solvent used in the solvent extraction method is not limited as long as the active ingredients can be sufficiently extracted into the extract. For example, an organic solvent, an aqueous solvent, or a mixed solvent of an organic solvent and an aqueous solvent can be used, but preferably a polar organic solvent, an aqueous solvent, or a mixed solvent containing a polar organic solvent or an aqueous solvent. The active ingredient that suppresses osteoclast differentiation contained in the osteoclast differentiation inhibitor of the present invention is considered to be efficiently dissolved in an aqueous solvent or a polar organic solvent that is a polar solvent. Therefore, particularly preferred extraction solvents are polar organic solvents or aqueous solvents.

Examples of the organic solvent include alcohol, acetone, benzene, ester, ethyl acetate, hexane, chloroform, and diethyl ether, and preferably a polar organic solvent. Examples of the polar organic solvent include alcohol, ester, ethyl acetate, hexane, chloroform, or diethyl ether. The alcohol is not limited as long as the active ingredients can be sufficiently extracted into the extract, but examples include alcohols with 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, isopropyl alcohol, and butyl alcohol. Examples include polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin.

The aqueous solvent is not limited as long as it contains water, and for example, water, physiological saline, or a buffer solution can be used. Examples of the buffer include phosphate buffer, sodium phosphate buffer, sodium carbonate buffer, citrate buffer, citrate phosphate buffer, acetate buffer, and Tris buffer. A preferred aqueous solvent is sodium phosphate buffer. The pH of the aqueous solvent is not particularly limited.

Furthermore, the extract used in the osteoclast differentiation inhibitor of the present invention can be extracted with a mixed solvent containing a polar organic solvent. The amount of polar organic solvent contained in the mixed solvent is not limited as long as the active ingredients can be sufficiently extracted into the extract, but the content of polar organic solvent relative to the total amount of extraction solvent is , for example, 50% by weight or more, 70% by weight or more, or 90% by weight or more. The extract used in the osteoclast differentiation inhibitor of the present invention can be extracted with a mixed solvent containing an aqueous solvent. The amount of the aqueous solvent contained in the mixed solvent is not limited as long as the active ingredients can be sufficiently extracted into the extract, but the content of the aqueous solvent relative to the total amount of the extraction solvent is, for example, , 50% by weight or more, 70% by weight or more, or 90% by weight or more.

When the extract is extracted by a solvent extraction method, the extraction temperature is not particularly limited as long as the effective ingredients can be sufficiently extracted into the extract, but it is between -50°C and 150°C. Preferably it is ℃. The lower limit of the extraction temperature is -25 °C or higher in some embodiments, 0 °C or higher in some embodiments, 5 °C or higher in some embodiments, 10 °C or higher in some embodiments, and 30 °C or higher in some embodiments. In some embodiments, the temperature is 50°C or higher. The upper limit of the extraction temperature is 120°C or less in some embodiments, 100°C or less in some embodiments, 80°C or less in some embodiments, 50°C or less in some embodiments, and 20°C or less in some embodiments. . The upper and lower limits of the temperature can be combined as appropriate.

Further, during extraction, it is preferable to carry out the extraction while stirring or shaking so as to improve the extraction efficiency. The extraction time can be appropriately determined depending on the part to be used, such as roots, stems, leaves, flowers, fruits, or seeds. In addition, the extraction time can be appropriately determined depending on the state of the plant, that is, whether it is fresh or dried, or if it is processed into a crushed or powdered state, the processing state. Furthermore, the extraction time can be determined as appropriate depending on the temperature of the extract liquid, or extraction conditions such as the presence or absence of stirring or shaking. The extraction time is usually 1 minute to 72 hours, preferably 1 hour to 48 hours, and most preferably 12 hours to 36 hours.

(steam distillation method)
The plant extract contained in the osteoclast differentiation inhibitor of the present invention can be extracted by steam distillation. The steam distillation method is a method in which steam is passed through a raw material packed in a column, and aromatic components distilled out along with the steam are condensed together with the steam. As the distillation means, any one of pressurized steam distillation, normal pressure steam distillation, and reduced pressure steam distillation can be employed.

(Squeezing method)
The squeezing method is a method of extracting extracts by applying physical pressure to plants. There are direct compression methods that are performed at room temperature, hot compression methods that are performed at high temperatures, and cold compression methods that are performed at low temperatures. The extract contained in the osteoclast differentiation inhibitor of the present invention can be extracted using any compression method.

(Supercritical extraction method)
The extract contained in the osteoclast differentiation inhibitor of the present invention can be extracted using a supercritical extraction method. The supercritical extraction method is a method of extracting extracts from specific plants using substances in a supercritical state. For example, carbon dioxide can be used as the substance in a supercritical state. Since carbon dioxide in a supercritical state has strong dissolving power, it is also commonly used for decaffeinating coffee or extracting flavoring and pharmaceutical ingredients from natural raw materials such as plants.

《Active ingredient》
The active ingredient contained in the osteoclast differentiation inhibitor of the present invention is contained in an extract extracted from a plant. Therefore, the plant contains a component that suppresses osteoclast differentiation, and the crushed product of the plant also contains the active component contained in the extract. Therefore, the crushed product of the above plant can also be used as an osteoclast differentiation inhibitor.
The active ingredient contained in the plant extract may be a fraction containing the active ingredient separated from the plant extract, or a purified active ingredient.

《Suppression of osteoclast differentiation》
The osteoclast differentiation inhibitor of the present invention suppresses the differentiation of osteoclast precursor cells into osteoclasts. Osteoclasts are multinucleated giant cells that originate from hematopoietic stem cells and are made up of several fused cells. Osteoclasts exhibit tartrate-resistant acid phosphatase (TRAP) activity in their cytoplasm. RANKL (receptor activator of NF-κB ligand), which is a signal for inducing differentiation into osteoclasts, binds to a receptor (RANK) on the membrane of osteoclast precursor cells, and differentiation into osteoclasts is initiated.
The osteoclast differentiation inhibitor of the present invention is an agent that inhibits differentiation of osteoclast precursor cells into osteoclasts. When the differentiation of osteoclast precursor cells into osteoclasts is promoted, the number of osteoclasts increases and bone resorption may be promoted. Excessive increase in bone resorption can lead to bone destruction and cause resorptive bone diseases such as osteoporosis, periodontal disease, and rheumatoid arthritis. The osteoclast differentiation inhibitor of the present invention is basically administered to patients with enhanced osteoclast differentiation. The causes of resorptive bone diseases include the promotion of differentiation of osteoclast precursor cells into osteoclasts, the promotion of bone resorption by osteoclasts, and the decrease in bone formation by osteoblasts. As described above, the osteoclast differentiation inhibitor of the invention is administered to patients with resorptive bone diseases in which osteoclast differentiation is accelerated.

"osteoporosis"
Bone tissue is maintained by destruction by osteoclasts (bone resorption) and formation by osteoblasts (bone formation). Under normal conditions, a balance of activity is maintained between osteoclasts and osteoblasts. When this balance shifts toward destruction (bone resorption), bone mass decreases, causing resorptive bone diseases such as osteoporosis. For example, osteoporosis is a disease in which bones become thin and brittle, causing fractures even under the stress of everyday life. Fractures in the elderly lead to bedriddenness and significantly reduce QOL. Osteoporosis is more common in post-menopausal women and older men, with approximately 30% of women in their 60s and over 40% in their 70s suffering from osteoporosis, and approximately 10% of men in their 60s suffering from osteoporosis. It has been reported that approximately 20% of people in their 70s have osteoporosis.

《Pharmaceutical composition》
The pharmaceutical composition for preventing or treating osteoporosis of the present invention contains the osteoclast differentiation inhibitor. The pharmaceutical composition of the present invention contains a crushed product or extract of a plant selected from the group consisting of coriander, bay, cardamom, chili pepper, pepper, sage, and clove, and can be used for the prevention or treatment of osteoporosis. can. Furthermore, the pharmaceutical composition of the present invention can be used as a pharmaceutical composition for inhibiting osteoclast differentiation.
The dosage form of the pharmaceutical composition of the present invention is not particularly limited, and may include oral dosage forms and parenteral dosage forms, with oral dosage forms being preferred. The oral preparations include, for example, solid or powder preparations such as fine granules, granules, tablets, capsules, and pills, and liquid preparations such as suspensions, emulsions, syrups, and extracts. can be mentioned. Examples of parenteral agents include injections.

The pharmaceutical composition of the present invention may consist of a ground plant product or a plant extract, or may contain a ground plant product or a plant extract. When the pharmaceutical composition of the present invention contains a ground plant product or a plant extract, it may contain other additives.

When the pharmaceutical composition of the present invention is an oral preparation, other additives include excipients, binders, disintegrants, emulsifiers, lubricants, fluidity promoters, diluents, preservatives, colorants, Flavoring agents, flavoring agents, stabilizers, humectants, preservatives, antioxidants, or suspending agents can be mentioned, and specific examples include gelatin, sodium alginate, starch, cornstarch, white sugar, lactose, Glucose, mannitol, carboxymethylcellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soybean lecithin, sucrose, fatty acid ester, talc, magnesium stearate, polyethylene glycol, magnesium silicate, silicic anhydride, or synthetic aluminum silicate. be able to.

When the pharmaceutical composition of the present invention is a parenteral preparation, other additives include water-soluble solvents such as physiological saline or Ringer's solution, water-insoluble solvents such as vegetable oil or fatty acid ester, glucose or sodium chloride, etc. Examples include tonicifying agents, solubilizing agents, stabilizers, preservatives, suspending agents, and emulsifying agents.

The pharmaceutical composition of the present invention can contain 90% by weight or more, 50% by weight or more, 10% by weight or more, or 1% by weight or more of a crushed plant product or extract.

The amount of administration or intake of the pharmaceutical composition of the present invention can be adjusted appropriately depending on the dosage form and the age, sex, weight, and severity of symptoms of the subject to whom the pharmaceutical composition is administered or Preferably, the amount is such that when ingested, the onset of osteoporosis can be prevented, or osteoporosis that has developed can be alleviated or treated. Specifically, the amount of the plant aqueous solvent extract added is 0.01 to 1000 mg/kg body weight/day, preferably 0.1 to 750 mg/kg body weight/day, and more preferably 1 to 500 mg/day. kg body weight/day, more preferably 5-400 mg/kg body weight/day, even more preferably 10-300 mg/kg body weight/day, even more preferably 15-200 mg/kg body weight/day, or most preferably 20-150 mg/kg. It can be weight/day. Of course, the above administration method is just an example, and other administration methods may be used. The administration method, dosage, administration period, administration interval, etc. of a pharmaceutical composition to humans are preferably determined by controlled clinical trials.

The pharmaceutical composition of the present invention can be administered to humans, but the subject of administration may also be animals other than humans, such as birds, pigs, horses, and the like.

The pharmaceutical composition of the present invention can be a pharmaceutical composition for preventing or treating osteoporosis. The pharmaceutical composition includes pharmaceuticals and quasi-drugs. Examples of pharmaceuticals include crude drug preparations and Chinese herbal preparations. Examples of quasi-drugs include nutritional drinks and health medicines containing crude drugs.

《Food composition》
The food composition for preventing or treating porosis of the present invention contains the osteoclast differentiation inhibitor. The food composition of the present invention contains a crushed product or extract of a plant selected from the group consisting of coriander, bay, cardamom, chili pepper, sage, pepper, and clove, and can be used for the prevention or treatment of osteoporosis. can. Furthermore, the food composition of the present invention can be used as a food composition for inhibiting osteoclast differentiation.

Examples of foods include freshly prepared foods such as salads; heated foods such as steak, pizza, and hamburgers; stir-fried foods such as stir-fried vegetables; tomatoes, green peppers, celery, bitter gourd, carrots, potatoes, and asparagus. vegetables such as vegetables and cooked products made from these vegetables; confectionery such as cookies, bread, biscuits, hardtack, cakes, rice crackers, yokan, puddings, jellies, ice creams, chewing gum, crackers, chips, chocolates and candies; udon noodles, Noodles such as pasta and soba; Fish paste products such as kamaboko, ham, and fish sausage; Dairy products such as cheese, cream, and butter; Miso, soy sauce, dressing, ketchup, mayonnaise, soup stock, noodle soup, and curry powder , seasonings such as mirin and roux; soybean foods such as tofu; processed agricultural and marine products such as furikake, tsukudani, and cereals; and konnyaku.

Beverages include, for example, coffee drinks; cocoa drinks; vegetable juices obtained from the above-mentioned vegetables; fruit juice drinks such as grapefruit juice, orange juice, grape juice, and lemon juice; tea drinks such as green tea, black tea, sencha, and oolong tea. Alcoholic beverages such as beer, wine (red wine, white wine, sparkling wine, etc.), sake, plum wine, low-malt beer, whiskey, brandy, shochu, rum, gin, and liqueurs; Milk beverages; Soy milk beverages; Liquid foods; Also, sports drinks and the like can be mentioned.

Food or beverages include animal feed and beverages. Examples of target animals include primates such as humans, birds, pigs, and horses.

These foods or beverages may contain antioxidants, flavors, acidulants, colorants, emulsifiers, preservatives, seasonings, sweeteners, spices, pH adjusters, stabilizers, vegetable oils, animal oils, sugars, and sugars, if desired. Food additives and food materials such as alcohols, vitamins, organic acids, fruit juice extracts, vegetable extracts, grains, beans, vegetables, meat, seafood, etc. can be blended singly or in combination of two or more. The blending amounts of these food materials and food additives can be appropriately determined within a range that does not impair the purpose of the present invention.

These foods or beverages can be sterilized using common methods such as heat and pressure sterilization using retorts and autoclaves, batch sterilization, plate sterilization, electrical heat sterilization, microwave heat sterilization, and steam sterilization such as injection and infusion. Sterilization treatment can be performed.

Foods and beverages include functional foods (drinks) and health foods (drinks). As used herein, "health food (beverage)" refers to a food or drink that has or can be expected to have some effect on health, and "functional food (beverage)" refers to Among "health foods (drinks)," there are foods and beverages that are designed and processed to fully exhibit bioregulatory functions (i.e., functions to prevent the onset of osteoporosis, or to alleviate or treat osteoporosis symptoms). means. Functional foods and health foods can be in the form of granules, solids, liquids, capsules, gels, or tablets.

EXAMPLES Hereinafter, the present invention will be specifically explained with reference to Examples, but these are not intended to limit the scope of the present invention.

《Preparation Examples 1 to 5》
In this preparation example, coriander seeds (preparation example 1), bay leaves (preparation example 2), chili pepper fruit (preparation example 3), pepper seeds (preparation example 4), clove buds (preparation example 5), Extracts were prepared from cardamom fruit (Preparation Example 6) or a mixture of sage leaves, stems, and spikes (Preparation Example 7) using ethanol.
Approximately 1 g of each lyophilized powder was suspended in 50 mL of ethanol and extracted using a shaker for 30 minutes at room temperature. Thereafter, insoluble matter was removed by filtration, and the solvent was distilled off using a rotary evaporator. The resulting residue was dissolved in DMSO.

《Example 1》
In this example, the effect of coriander extract on osteoclast differentiation was investigated. RAW264 cells (precursor osteoclasts) were seeded in a 96-well culture plate using α-MEM medium. On day 0, RANKL (140 ng/mL) was added to induce osteoclast differentiation. At the same time, coriander extract (12.5, 25, or 50 μg/mL; DMSO final concentration 0.1%) was added and cultured for 5 days. Phosphatase chromogenic substrate pNPP (2 mg/mL) was added to the plate, incubated for 30 minutes at 37°C, and TRAP5b activity was measured by absorbance. As a negative control, one in which RANKL and coriander extract were not added, and as a positive control, one in which only RANKL was added. As shown in Figure 1, osteoclast differentiation was suppressed by coriander extract compared to the positive control.

《Example 2》
In this example, the effect of bay extract on osteoclast differentiation was investigated. The procedure of Example 1 was repeated, except that bay extract (12.5 or 25 μg/mL; DMSO final concentration 0.1%) was used. As shown in FIG. 2, osteoclast differentiation was suppressed by the bay extract compared to the positive control.

《Example 3》
In this example, the effect of chili pepper extract on differentiation into osteoclasts was investigated. The procedure of Example 1 was repeated, except that chili pepper extract (12.5, 25, or 50 μg/mL; DMSO final concentration 0.1%) was used. As shown in Figure 3, osteoclast differentiation was suppressed by chili pepper extract compared to the positive control.

《Example 4》
In this example, the effect of pepper extract on differentiation into osteoclasts was investigated. The procedure of Example 1 was repeated, except that pepper extract (12.5, 25, or 50 μg/mL; DMSO final concentration 0.1%) was used. As shown in FIG. 4, osteoclast differentiation was suppressed by the pepper extract compared to the positive control.

《Example 5》
In this example, the effect of clove extract on differentiation into osteoclasts was investigated. The procedure of Example 1 was repeated, except that clove extract (50 μg/mL; DMSO final concentration 0.1%) was used. As shown in Figure 5, osteoclast differentiation was suppressed by clove extract compared to the positive control.

《Example 6》
In this example, the effect of cardamom extract on differentiation into osteoclasts was investigated. The procedure of Example 1 was repeated, except that cardamom extract (25 or 50 μg/mL; DMSO final concentration 0.1%) was used. As shown in FIG. 6, osteoclast differentiation was suppressed by cardamom extract compared to the positive control.

《Example 7》
In this example, the effect of sage extract on differentiation into osteoclasts was investigated. The procedure of Example 1 was repeated, except that sage extract (12.5, 25, or 50 μg/mL; DMSO final concentration 0.1%) was used. As shown in FIG. 7, osteoclast differentiation was suppressed by sage extract compared to the positive control.

The osteoclast differentiation inhibitor of the present invention can be used to treat patients with enhanced osteoclast differentiation.

Claims (5)

An osteoclast differentiation inhibitor comprising a ground product or extract of a plant selected from the group consisting of coriander, bay, cardamom, chili pepper, pepper, sage, and clove. The osteoclast differentiation inhibitor according to claim 1, wherein the extract is an extract using a polar organic solvent, an aqueous solvent, or a mixed solvent containing a polar organic solvent or an aqueous solvent. The osteoclast according to claim 1 or 2, wherein the plant is coriander fruit, bay leaf, cardamom fruit, chili pepper fruit, pepper fruit, sage leaf, stem, or flower spike, or clove flower bud. Cell differentiation inhibitor. A pharmaceutical composition for preventing or treating osteoporosis, comprising the osteoclast differentiation inhibitor according to claim 1 or 2. A food composition for preventing or treating osteoporosis, comprising the osteoclast differentiation inhibitor according to claim 1 or 2.