JP2018090528A - Articular cartilage degradation inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide pharmaceuticals, foods and the like which suppress the decomposition of articular cartilage and are useful for the prevention or improvement of joint disorders caused by the decomposition of articular cartilage. SOLUTION: An articular cartilage decomposition inhibitor obtained by combining ellagic acid or a derivative thereof or a salt thereof and catechins. [Selection diagram] None

Description

  The present invention relates to an articular cartilage degradation inhibitor that inhibits articular cartilage degradation.

  In recent years, the population affected by bone and joint disease, which is one of the factors that impair the healthy life expectancy of the elderly, has been increasing. Many osteoarthritis diseases tend to increase in number with the onset of aging, and with the recent aging of the population, a rapid increase in the affected population is expected in the future.

  Osteoarthritis, a bone and joint disease, is a progressive disease characterized by destruction, degeneration, and disappearance of articular cartilage. For example, knee osteoarthritis is a disease in which cartilage and meniscus are gradually worn down, making joint deformation, pain generation, and bending and stretching of joints difficult. The total number of patients with knee osteoarthritis in Japan is estimated to be about 8 million, which is far greater than rheumatoid arthritis, which is one of the same bone and joint diseases.

  At present, pharmacotherapy for osteoarthritis is a symptomatic treatment for the purpose of pain relief, and there are few established treatments for suppressing the progression of the disease itself. Further, many conventional therapeutic drugs have problems such as side effects, and safer and more effective preventive / ameliorating drugs are desired.

  Osteoarthritis is caused by the destruction of cartilage tissue. Cartilage tissue is composed of chondrocytes and the extracellular matrix surrounding them, and the main components of the articular cartilage extracellular matrix (hereinafter also simply referred to as the cartilage matrix) are type II collagen and aggrecan (proteoglycan). Collagen contributes to cartilage morphology maintenance, while aggrecan contributes to cartilage viscoelasticity. In a healthy person's cartilage tissue, the metabolic balance between collagen and aggrecan production and cartilage matrix degradation by cartilage matrix degrading enzymes is maintained appropriately. On the other hand, in osteoarthritis patients, since the degradation and degeneration of articular cartilage matrix is observed in the early stage of onset, it is considered that the metabolic balance of cartilage matrix tends to be degraded.

  As enzymes that degrade cartilage matrix, metalloproteases belonging to the ADAMTS (A Disintegrin And Metalloproteinase with ThoromboSpondin motifs) family and metalloproteases belonging to the MMP (Matrix Metalloproteinase) family are known. These families include the enzyme aggrecanase that degrades aggrecan and the enzyme MMP that degrades collagen and aggrecan.

  The onset and progression of osteoarthritis is thought to involve proteolytic enzymes responsible for the degradation of cartilage matrix centered on aggrecan and collagen. Recent research suggests that cartilage matrix destruction is first preceded by enzymatic degradation and disappearance of aggrecan, followed by enzymatic degradation of collagen, and the presence of aggrecan protects collagen from degradation. (For example, refer nonpatent literature 1).

  On the other hand, ellagic acid used as a food additive (antioxidant) has an action of inhibiting the activity of gelatinase, one of the enzymes that degrade collagen, and is useful for the prevention or improvement of arthritis. (Patent Document 1) and the activity of inhibiting aggrecanase, which is an aggrecan-degrading enzyme, have been reported to effectively suppress aggrecan degradation of cartilage matrix (Patent Document 2). In addition, catechins derived from green tea have been reported to have an action of inhibiting the degradation of bovine and human cartilage proteoglycans and type II collagen, and are useful for the prevention and improvement of arthritis by suppressing cartilage destruction (non-) Patent Document 2).

  However, there is no report on the inhibition of articular cartilage destruction by the combination of ellagic acid and catechins, and the effect of the combination is not known at all.

JP 2005-47817 A JP 2013-245168 A

Linda Troeberg, et al., Biochimica et Biophysica Acta 1824, 2012, 133-145 Adcocks C et al., J Nutr. 132 (3), 2002, 341-346

  The present invention relates to providing pharmaceuticals, foods, and the like that are useful for preventing or improving joint damage by inhibiting the degradation of articular cartilage and inhibiting articular cartilage degradation.

  The present inventors have examined a material that suppresses the degradation of articular cartilage tissue, and when used in combination with ellagic acid and catechins, exhibits an excellent cartilage degradation inhibitory effect, which prevents joint damage or It was found to be extremely useful for improvement.

That is, the present invention relates to the following 1) to 6).
1) An articular cartilage degradation inhibitor obtained by combining ellagic acid or a derivative thereof or a salt thereof and catechins.
2) An articular cartilage degradation inhibitor comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.
3) A food for inhibiting articular cartilage degradation comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.
4) A preventive or ameliorating agent for osteoarthritis comprising a combination of ellagic acid or a derivative thereof or a salt thereof and catechins.
5) A preventive or ameliorating agent for osteoarthritis comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.
6) A food for preventing or ameliorating osteoarthritis comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.

  ADVANTAGE OF THE INVENTION According to this invention, a pharmaceutical, a quasi-drug, or a food useful for suppression of joint cartilage destruction or prevention or improvement of joint damage caused by joint cartilage destruction can be provided.

The graph which shows articular cartilage degradation inhibitory activity. The graph which shows articular cartilage degradation inhibitory activity (concentration dependence).

  The ellagic acid used in the present invention is a compound represented by the following formula (1).

  In the present invention, ellagic acid derivatives include methylated or acetylated ellagic acid, such as 3,4-di-o-methylellagic acid, 3,3′-di-o-methylellagic acid, 3 3,3′4-tri-o-methylellagic acid, 3,3 ′, 4,4′-tetra-o-methyl-5-methoxyellagic acid, 3-o-ethyl-4-o-methyl-5-hydroxyellag An acid etc. are illustrated suitably.

  Ellagic acid and ellagic acid derivatives may take the form of salts or hydrates thereof. Preferred examples of the salt include alkali metal salts such as sodium salt and potassium salt, and preferred examples of the hydrate include dihydrate.

Ellagic acid or a derivative thereof used in the present invention may be chemically synthesized by a known synthesis method, extracted from a natural product, or a commercially available product may be used. Examples of commercially available ellagic acid include pomegranate ellagic acid (Sabinsa Japan Corporation), ellagic acid dihydrate (Wako Pure Chemical Industries, Ltd.), ellagic acid (Kanto Chemical Co., Ltd.) and the like.
As a method for extracting ellagic acid from a natural product, for example, a dried natural product containing ellagic acid is digested by an acidic sulfite method, then immersed in an alkaline aqueous solution, and the resulting immersion solution is collected. And a method for collecting and purifying a precipitate containing ellagic acid by adding an acid to the immersion liquid.

The “catechins” in the present invention are non-epimeric catechins such as catechin, catechin gallate, gallocatechin and gallocatechin gallate, and epicatechins such as epicatechin, epigallocatechin, epicatechin gallate and epigallocatechin gallate. In the present invention, at least one non-polymer catechin may be contained. In the present invention, the content of catechins is defined based on the total amount of the eight non-polymer catechins.
Since the catechins used in the present invention are generally contained in a tea extract extracted from tea leaves, a concentrate thereof or a purified product thereof, those obtained from these are preferably used. Here, “tea extract” refers to a product extracted from tea leaves using water or a hydrophilic organic solvent, and has not been concentrated or purified. In addition, alcohol, such as ethanol, can be used as a hydrophilic organic solvent, for example. In addition, a kneader or a column can be used for extraction. The “tea extract concentrate” refers to a product obtained by removing at least part of the solvent from the tea extract extracted from tea leaves with water or a hydrophilic organic solvent to increase the concentration of catechins. It can be prepared by the methods described in JP-A-59-219384, JP-A-4-20589, JP-A-5-260907, JP-A-5-306279, and the like. Further, the “purified tea extract” refers to a product obtained by treating a tea extract or a concentrate thereof with a solvent or an adsorbent to increase the purity of catechins in the solid content. No. 147508, JP-A 2007-282568, JP-A 2006-160656, JP-A 2008-079609 and the like.
Examples of tea leaves used for extraction include tea trees selected from the genus Camellia, such as C. sinensis and C. assamica, Yabuki species, or hybrids thereof. Tea leaves can be broadly classified into non-fermented tea, semi-fermented tea, and fermented tea depending on the processing method. Examples of non-fermented tea include green tea such as sencha, bancha, mochi tea, kettle tea, stem tea, stick tea, and bud tea. Examples of the semi-fermented tea include oolong tea such as iron kannon, color type, golden katsura, and martial arts tea. Furthermore, examples of fermented tea include black teas such as Darjeeling, Assam, Sri Lanka and the like. These can be used alone or in combination of two or more.

  The articular cartilage degradation inhibitor comprising a combination of ellagic acid or a derivative thereof or a salt thereof (hereinafter referred to as “ellagic acid”) and catechins according to the present invention is formulated as an appropriate amount of each effective amount. It may be formulated into a single dosage form in the ratio, or may be a kit that can be used separately or separately at intervals, with the formulation containing each effective amount alone. .

  In the present invention, the ratio of the combination of ellagic acids and catechins can be appropriately selected. For example, the catechins are preferably 0.01 parts by mass or more, more preferably 0 with respect to 1 part by mass of the ellagic acids. 0.1 parts by mass or more, more preferably 0.2 parts by mass or more, more preferably 0.5 parts by mass or more, and preferably 100 parts by mass or less, more preferably 10 parts by mass or less, more preferably 5 parts by mass. Hereinafter, it is more preferably 2 parts by mass or less. Moreover, Preferably it is 0.01-100 mass parts, More preferably, it is 0.1-10 mass parts, More preferably, it is 0.2-5 mass parts, More preferably, it is 0.5-2 mass parts.

As shown in the Examples below, the combination of ellagic acids and catechins significantly suppressed the destruction of IL-1 induced articular cartilage, and the effect was obtained when ellagic acid and catechins were each administered alone. It can be said that it is a synergistic effect that exceeds the additive effect.
Therefore, a combination of ellagic acids and catechins can be used to suppress articular cartilage degradation, can be an articular cartilage degradation inhibitor, and can be used to produce an articular cartilage degradation inhibitor.

  Cartilage is an elastic supporting tissue composed of cartilage tissue and perichondrium, and articular cartilage is cartilage that thinly covers the ends of two bones that form a joint. It is known that joint damage such as osteoarthritis, rheumatoid arthritis, osteoarthritis, etc. is caused by the degradation and destruction of articular cartilage. It is useful for the prevention or amelioration of joint disorders caused by, for example, osteoarthritis. Here, osteoarthritis refers to a disease characterized by degeneration and wear of articular cartilage, hardening of subchondral bone, and proliferative change.

  Therefore, the combination of the ellagic acids and catechins of the present invention can be used to prevent or ameliorate osteoarthritis, can be a preventive or ameliorating agent for osteoarthritis, and can be used to prevent or improve osteoarthritis. Can be used to produce the agent.

  In addition, "use" when using a combination of the above-mentioned ellagic acids and catechins for preventing or improving articular cartilage degradation and osteoarthritis can be used in humans or non-human animals, It may be therapeutic or non-therapeutic. Here, “non-therapeutic” means a concept that does not include medical practice, that is, a concept that does not include a method for operating, treating, or diagnosing a person, more specifically, a doctor or a person who has received instructions from a doctor It is a concept that does not include a method for performing surgery, treatment, or diagnosis on the subject.

The above-mentioned articular cartilage degradation inhibitor, osteoarthritis prevention or amelioration agent is an articular cartilage degradation suppression, osteoarthritis for preventing or ameliorating oral drugs, quasi drugs, supplements or foods, or It can be a material or a preparation for blending them.
In addition to general foods and drinks, the above foods are based on the concept of inhibiting the degradation of articular cartilage, preventing or improving osteoarthritis, etc. Food, food for the sick, food with nutritional function, and the like are included.

The above-mentioned pharmaceuticals (including quasi drugs) containing ellagic acids and catechins can be administered in any dosage form. Examples of the dosage form include oral administration such as tablets, capsules, granules, powders, syrups, etc. or parenteral administration such as injections, suppositories, inhalants, transdermal absorption agents, external preparations, etc. The form is oral administration.
In order to prepare such pharmaceutical preparations of various dosage forms, the ellagic acids and catechins of the present invention can be used alone, or other pharmaceutically acceptable excipients, binders, extenders, disintegrants. , Surfactants, lubricants, dispersants, buffers, preservatives, flavoring agents, fragrances, coating agents, carriers, diluents, and the like can be used in appropriate combinations.

The form of the above-mentioned food containing ellagic acids and catechins includes various food compositions such as breads, cakes, noodles, confectionery, jelly, frozen foods, ice creams, dairy products, beverages, Examples include the same forms (tablets, capsules, syrups, etc.) as the above-mentioned oral administration preparations.
In order to prepare various forms of food, the ellagic acids and catechins of the present invention alone or other food materials, solvents, softeners, oils, emulsifiers, preservatives, fragrances, stabilizers, colorants , Antioxidants, humectants, thickeners, and the like can be used in appropriate combinations.

  The content of ellagic acids in the above-mentioned pharmaceuticals (including quasi drugs) and foods varies depending on the use form, but is usually 0.02% by mass or more, preferably 2% by mass or more of the total mass of the preparation. And 100% by mass or less, preferably 99.8% by mass or less. Moreover, it is 0.02-100 mass%, Preferably it is 2-99.8 mass%. The content of catechins is 0.002% by mass or more, preferably 0.2% by mass or more, and 100% by mass or less, preferably 98% by mass or less, based on the total mass of the preparation. Moreover, it is 0.002-100 mass%, Preferably it is 0.2-98 mass%.

  When the combination of ellagic acids and catechins of the present invention is used as a pharmaceutical or food, or in combination with a pharmaceutical or food, the dosage may vary according to the patient's condition, weight, sex, age or other factors. In the case of oral administration, the daily dose per adult is usually preferably 0.025 mg or more, more preferably 2.8 mg or more, and preferably 14.4 g or less, more preferably ellagic acids. 3 g or less. Further, it is preferably 0.025 mg to 14.4 g, more preferably 2.8 mg to 3 g. The catechins are preferably 0.0024 mg or more, more preferably 0.22 mg or more, and preferably 2.7 g or less, more preferably 1.5 g or less. Moreover, Preferably it is 0.0024 mg-2.7 g, More preferably, it is 0.22 mg-1.5 g.

Moreover, although the said formulation can be administered according to arbitrary administration schedules, it is preferable to divide once to several times a day, and to administer continuously for several weeks to several months.
The subject of administration or ingestion is not particularly limited as long as it is an animal that needs or desires it, but a human who needs or desires prevention or improvement of joint disorders, more preferably osteoarthritis. Is mentioned.

Regarding the above-described embodiment, the following aspects are further disclosed in the present invention.
<1> An articular cartilage degradation inhibitor comprising a combination of ellagic acid or a derivative thereof or a salt thereof and catechins.
<2> An articular cartilage degradation inhibitor comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.
<3> A food for inhibiting articular cartilage degradation comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.
<4> A preventive or ameliorating agent for osteoarthritis comprising a combination of ellagic acid or a derivative thereof or a salt thereof and catechins.
<5> A preventive or ameliorating agent for osteoarthritis comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.
<6> A food for preventing or ameliorating osteoarthritis comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.

<7> A combination of ellagic acid or a derivative thereof or a salt thereof and catechins for use in inhibiting articular cartilage degradation.
<8> A combination of ellagic acid or a derivative thereof or a salt thereof and catechins for producing an articular cartilage degradation inhibitor.
<9> A method for inhibiting articular cartilage degradation, comprising administering or ingesting an effective amount of ellagic acid or a derivative thereof or a salt thereof in combination with catechins.

<10> A combination of ellagic acid or a derivative thereof or a salt thereof and catechins for use in the prevention or improvement of osteoarthritis.
<11> A combination of ellagic acid or a derivative thereof or a salt thereof and catechins for producing an agent for preventing or improving osteoarthritis.
<12> A method for preventing or improving osteoarthritis, comprising administering or ingesting an effective amount of ellagic acid or a derivative thereof or a salt thereof in combination with catechins.

<13> In the above <7> or <10>, the use is a non-therapeutic use.
<14> In the above <9> or <12>, the method is a non-therapeutic method.
<15> In the above <1> to <14>, the ratio of the combination of ellagic acids and catechins is preferably 0.01 parts by mass or more, more preferably catechins with respect to 1 part by mass of ellagic acids, for example. 0.1 parts by mass or more, more preferably 0.2 parts by mass or more, more preferably 0.5 parts by mass or more, and preferably 100 parts by mass or less, more preferably 10 parts by mass or less, more preferably 5 parts by mass. Part or less, more preferably 2 parts by weight or less. Moreover, Preferably it is 0.01-100 mass parts, More preferably, it is 0.1-10 mass parts, More preferably, it is 0.2-5 mass parts, More preferably, it is 0.5-2 mass parts.

EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited to this.
<Preparation of cartilage tissue and test medium>
(1) Extraction of tissue A femur (6 months old) with no damage to the porcine knee joint was purchased from Funakoshi. The porcine knee joint capsule on the femoral condyle side was incised to expose the cartilage, and a cylindrical piece of cartilage was removed with a punch biopsy having a diameter of 3 mm. The disc-shaped tissue weight obtained by scraping off the tissue adhering to the bone side opposite to the cartilage surface on the back side of the scalpel was measured.

(2) Tissue culture After weighing, each well was placed in a 96-well microplate containing D-PBS (-) (Life Technologies, 14190-144) 1 volume% PS (Penicillin Streptomycin: Life Technologies, 15140-122). One piece was dipped per well. After switching from PBS (−) 1 volume% PS to DMEM (Dulbecco's modified Eagle medium), 10 volume% FBS (fetal bovine serum), 1 volume% PS (200 μl / well), DMEM, 1 volume% FBS, 1 5-6 days 37 ° C. by switching on the volume% PS, and subjected to assay by culturing at 5 vol% CO _2. The medium was changed every 2-3 days.

(3) Preparation of test medium The test medium was prepared by adding various samples to DMEM, 1 vol% PS (0 vol% FBS). The added samples are as follows.
1) Control group 2) Ellagic acid (100 ng / ml)
3) Catechin (100 ng / ml)
4) Ellagic acid (100 ng / ml) + catechin (100 ng / ml)
* Elagic acid: Wako Pure Chemical Industries, Ltd. * Catechins: 8 types <Catechin (C), Epicatechin (EC), Gallocatechin (GC), Epigallocatechin (EGC), Epicatechin gallate (ECg), Catechin gallate (Cg) ), Epigallocatechin gallate (EGCg), and gallocatechin gallate (GCg)> are used (100 ng / ml each, catechin total 800 ng / ml).

Test Example 1 Articular cartilage degradation inhibitory activity (1) Sample addition and degradation stimulation Cartilage tissue was allocated to each group so that there was no difference in the average tissue weight between groups, and the corresponding test medium (200 μl prepared in (3) above) / well) and cultured. For tissue degradation stimulation, rh IL-1α (Recombinant Human IL-1α, R & D Systems) was added at a final concentration of 80 ng / ml after 1 hour of culture in the test medium. After a further 24 hours of culture, the medium was collected.

(2) Measurement of degradation index Glycosaminoglycan (GAG) released from the aggrecan in the cartilage tissue into the medium was measured with a Blyscan Glycosaminoglycan Assay Kit (Biocolor), and the GAG amount / cartilage tissue weight was determined. It was used as an index.

(3) Results The results are shown in FIG.
In the group to which ellagic acid and catechins were added in combination, cartilage degradation was far more suppressed than in the group to which ellagic acid and catechins were added alone, and the effect was synergistic.

Test Example 2 Articular cartilage degradation inhibitory activity (concentration-dependent effect)
As an additive sample, a combination of ellagic acid (20,100,500 ng / ml) and catechins (20,100,500 ng / ml) at the same concentration was used, and cartilage degradation inhibitory activity was measured in the same manner as in Test Example 1. .
The result is shown in FIG. From FIG. 2, the effect of suppressing cartilage degradation alone was not observed at a concentration of 500 ng / ml or less for both ellagic acid and catechins. However, it was confirmed that the group using both from 100 ng / ml synergistically suppresses cartilage degradation than the group added alone.

Claims (8)

  An articular cartilage degradation inhibitor comprising a combination of ellagic acid or a derivative thereof or a salt thereof and catechins.   An articular cartilage degradation inhibitor comprising ellagic acid or a derivative thereof or a salt thereof and catechin as active ingredients.   A food for inhibiting articular cartilage degradation comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.   A prophylactic or ameliorating agent for osteoarthritis comprising a combination of ellagic acid or a derivative thereof or a salt thereof and catechins.   A preventive or ameliorating agent for osteoarthritis comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.   A food for preventing or improving osteoarthritis, comprising ellagic acid or a derivative thereof or a salt thereof and catechins as active ingredients.   The rate of the combination of ellagic acid or a derivative thereof or a salt thereof and catechins is 0.01 to 100 parts by mass of catechins with respect to 1 part by mass of ellagic acids. An agent or a food for inhibiting articular cartilage degradation according to claim 3.   The osteoarthritis according to claim 4 or 5, wherein the proportion of ellagic acid or a derivative thereof or a combination thereof and catechins is 0.01 to 100 parts by mass of catechins with respect to 1 part by mass of ellagic acids. The preventive or ameliorating agent according to claim 6, or the food for preventing or improving osteoarthritis according to claim 6.

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