JP6547183B2 - Composition for prevention or treatment of osteoarthritis comprising triterpene as an active ingredient - Google Patents

Description

  The present invention relates to a composition for preventing or treating a disease associated with a cartilage abnormality such as osteoarthritis, which contains a triterpene as an active ingredient. The present invention also relates to a composition for promoting cartilage collagen metabolism, which comprises triterpene as an active ingredient.

Vertebrates, including humans, are at risk of developing musculoskeletal syndrome (also called locomotive syndrome) due to aging and lifestyle. Musculoskeletal syndrome has been defined by the Japanese Association of Orthopedic Surgeons as "a state in which the risk of needing care increases due to the decline or failure of the musculoskeletal system". Almost all joints in vertebrates have cartilage, and cartilage is important for providing joint flexibility, compressibility and other smoothness of movement within the joint, and cartilage flexibility causes joint flexibility. It causes many symptoms, including loss of compressibility, inflammation of the joints and tissues surrounding the joints, and pain. Examples of such conditions include osteoarthritis, rheumatoid arthritis, degenerative spondylosis and the like. In addition, motor disorders due to traumatic factors such as injuries are also included in the motor organ syndrome.
Osteoarthritis tends to develop in weight-bearing joints such as knees, hips and ankles. It can be caused by traumatic joint damage, but most are due to wear and degeneration and aging of the cartilage with aging.
Cartilage is a supporting organ consisting of chondrocytes and a matrix. The main components of the matrix are type II collagen and proteoglycans, and type II collagen is complexed with proteoglycans to constitute a fiber network. Cartilage wear means that the cartilage is gradually worn out by physical stimulation such as exercise, and cartilage degeneration is that the matrix component is damaged such as breakage due to physical stimulation. Cartilage aging is said to be the decrease in the ability to produce matrix by chondrocytes and the accumulation of changes at the molecular level of the matrix. Changes at the molecular level include the addition of AGE (final glycation reactant) to type II collagen and the reduction of molecular weight of aggrecan, which is the main component of proteoglycan. During AGE addition to type II collagen, accumulation of an AGE called pentosidine, in which pentoses cross-link lysine and arginine residues of amino acids constituting collagen, makes the tissue hard and brittle, resulting in reduced elasticity of the cartilage matrix. The half life of cartilage collagen is estimated to be over 100 years, and AGE accumulation is a major problem for osteoarthritis.
In chondrocytes, type II collagen is synthesized as procollagen containing propeptides PIINP and PIICP at the N-terminus and C-terminus, respectively. Procollagen is digested by a specific enzyme to become mature type II collagen and constitutes a cartilage matrix, and PIINP and PIICP are released into the blood. Type II collagen has type IIA and type IIB by alternative splicing, and type IIB N-terminal procollagen is PIINP, and type IIA is PIIANP including PIINP containing a cysteine-rich globular domain consisting of 69 amino acids. PIIANP levels are reduced in the sera of osteoarthritis-affected individuals (Non-patent Document 1), and PIIANP is regarded as a useful marker for osteoarthritis as a so-called "cartilage synthesis marker".
Moreover, in osteoarthritis, it is known that the cartilage matrix degrading enzymes MMP-1, MMP-8, and MMP-13 are upregulated in the surface layer of the cartilage (Non-patent Document 2). C2C is a neoepitope of type II collagen released into the blood by enzymatic cleavage of type II collagen, and C2C is a so-called "cartilage degradation marker" and is used for osteoarthritis such as osteoarthritis and rheumatism It is considered as a useful marker (patent document 1).
Currently, drug treatment of diseases related to cartilage abnormalities such as osteoarthritis is only a symptomatic treatment to relieve pain and swelling caused by inflammation, and symptomatic treatment to prevent deterioration of joint function, and is a more effective treatment. Medicine is desired.

Triterpenes are a group of compounds widely distributed in the plant world, and are a group of compounds classified into oleanane type terpenes and ursan type triterpenes. The oleanane triterpenes include maslinic acid, oleanolic acid and the like, and the ursan triterpenes include ursolic acid, corosolic acid, tolmentic acid and the like.
Plant-derived triterpenes have recently been attracting attention as functional components having physiological activity, and for maslinic acid, they have large-bowel cancer suppressive action (non-patent document 1), skin whitening (patent document 2), osteoclast differentiation inhibition and bone The composition has an absorption activity effect (patent document 3), an obesity preventing effect (patent document 4), indigestible dextrin, corosolic acid, maslinic acid and tolmentic acid as active ingredients, and suppresses the increase of blood glucose level (patent document 5) With respect to oleanolic acid, anti-caries (prevention of dental caries) (Patent Document 6), anti-cancer effect (Patent Document 7), proton pump inhibitory action (Patent Document 8) and the like for ursolic acid have been reported.
However, there is no attempt or suggestion on the prevention or treatment of diseases associated with cartilage abnormalities such as osteoarthritis using triterpenes.

Special Table 2006-501479 International Publication 2002/043736 JP 2014-141444 International Publication 2003/011267 Japanese Patent Application Publication No. 2006-34967 Japanese Patent Application Laid-Open No. 1-290619 JP-A-8-119866 JP 2012-25719

Rousseau JC et al., Osteoarthritis Cartilage. 2004 Jun; 12 (6): 440-7 Okazaki Ken, Iwamoto Yukihide, Fukuoka Medical Journal 103 (9): 175-181, 2012 Juan ME et al., J Nutr., 136 (10), 2553-2557, 2006

  An object of the present invention is to provide a composition for preventing or treating a disease associated with cartilage abnormality such as osteoarthritis. Another object of the present invention is to provide a composition for promoting cartilage collagen metabolism.

The present inventors have found that the consumption of triterpenes derived from plants raises the cartilage synthesis marker and the cartilage degradation marker together and reduces inflammation markers, thereby promoting cartilage collagen metabolism. By promoting cartilage collagen metabolism, it is possible to delay the progression of symptoms of diseases associated with cartilage abnormalities and alleviate the symptoms. Furthermore, it has been found that pain is alleviated in patients suffering from osteoarthritis, and the QOL of the patient is improved, and it can be used as a composition for preventing or treating osteoarthritis.
That is, the present invention is as follows.
[1] A composition for the prevention or treatment of osteoarthritis comprising a triterpene or a pharmacologically acceptable salt thereof as an active ingredient.
[2] A composition for promoting cartilage collagen metabolism, comprising a triterpene or a pharmacologically acceptable salt thereof as an active ingredient.
[3] The composition according to [1] or [2] above, wherein the triterpene is an oleanane type or ursan type triterpene.
[4] The composition according to [3] above, wherein the triterpene is an oleanane type triterpene selected from maslinic acid and oleanolic acid.
[5] The composition according to [3] above, wherein the triterpene is an ursan-type triterpene selected from ursolic acid and corosolic acid.

  The composition containing the triterpene of the present invention as an active ingredient induces the synthesis and degeneration of type II collagen by chondrocytes and the degradation of type II collagen in the surface layer of aged cartilage to promote cartilage metabolism and simultaneously inflammation And since it can relieve pain and improve the patient's physical quality of life, it is effective in the prevention or treatment of diseases associated with cartilage abnormalities such as osteoarthritis.

It is an HPLC analysis chart of the liquid containing triterpenes obtained from olive fruit. The peak of A is maslinic acid and the peak of B is oleanolic acid.

In the present invention, a triterpene is a compound having a carbon number of 30 as a basic skeleton, and a pentacyclic triterpene is mentioned as a typical example thereof. Here, a pentacyclic triterpene is one kind of triterpenes, and is a pentacyclic compound consisting of six isoprene units, which has a carbon number of 30 as a base, but is rearranged, oxidized, Also included are those which are eliminated or alkylated and the carbon number fluctuates.
These can be obtained from natural plants or artificially. Moreover, a commercial item can also be utilized suitably. Pentacyclic triterpenes are generally classified by their backbone. For example, oleanane type triterpenes, ursan type triterpenes, lupan type triterpenes, popane type triterpenes, seratane type triterpenes, Friedelan type triterpenes, taraxelan type triterpenes, taraxastane type triterpenes, multifloran type triterpenes, jaranican type triterpenes, etc. may be mentioned.
Among the pentacyclic triterpenes, for example, maslinic acid, oleanolic acid, and ursolic acid, corosolic acid, tolmentic acid and the like can be mentioned as representative examples of oleanane type triterpenes.

  Maslinic acid is contained in olive, jujube, almond, banaba leaf, sage, almond, apple, cranberry, karin, banaba leaf, cranberry and the like. Oleanolic acid is olive, grape, beet, jujube, almond, banaba leaf, olive leaf, sage, hawthorn, raspberry, karin, rosemary leaf, guava, shiso leaf, blueberry, prune, loquat, pomegranate, lemon balm, basil, rose It is included in hips, oysters, slivers, etc. Ursolic acid is olive (leaf), almond, apple, cranberry, hawthorn, raspberry, cullin, rosemary leaf, guava, shiso leaf, blueberry, prune, loquat, pomegranate, lemon balm, basil, rosehip, oyster, semenuri, berry include. Corosolic acid is contained in karin, almonds, banaba leaves, cranberries, loquat leaves, perilla leaves and the like. Tolmentic acid is contained in clove, strawberry and the like.

  In the present invention, the triterpene is preferably selected from oleanane type triterpenes and ursane type triterpenes, more preferably selected from maslinic acid, oleanolic acid, ursolic acid and corosolic acid, and most preferably maslinic acid and / or oleanolic acid.

  These triterpenes can be obtained from the above-mentioned plants by extraction and purification by known methods, and can also be obtained by chemical synthesis. The triterpene may be a single component or a mixture of several components. They can also be used in the form of pharmacologically acceptable salts and / or derivatives.

For example, a lower alcohol (ethanol, methanol, n-propanol, isopropanol, n, etc.) which can be extracted from dried, semi-dried or undried olive fruits and olive oil mill produced in the oil extraction process as a starting material and triterpenes such as maslinic acid can be extracted -Extract triterpenes with butanol etc.) or its hydrous alcohol, saponify and neutralize if necessary, and use octadecyl silica (ODS), silica gel, synthetic adsorbent etc. as adsorbent, Fractionation and purification can be performed to obtain a mixture of triterpenes containing maslinic acid and oleanolic acid as main components.
When using other plants as a starting material, triterpene can be obtained by the method suitable for the plant used as a raw material. Moreover, the kind of the component of the triterpene obtained and its composition ratio depend on the plant used as a raw material.

  The dose of the composition of the present invention varies depending on the age, sex, degree of symptoms and administration form of the patient to be administered, but in an adult, the amount of triterpene per day is preferably 0.1 mg / kg or more. There is no upper limit of the dose, but it is not preferable from the viewpoint of product cost if it exceeds 50 mg / kg.

  The composition of the present invention can be formulated into an optional form together with an auxiliary agent to be a pharmaceutical which can be orally or parenterally administered. For example, oral dosage forms can be tablets, oral fast disintegrating tablets, capsules, solid dosage forms such as granules, granules, liquid dosage forms such as syrups and suspensions. As a parenteral dosage form, it is administered in the form of injection, eye drop, nasal drop, patch, suppository, external preparation for skin. Pharmaceuticals also include quasi drugs.

  In the case of solid dosage forms, appropriate amounts of various additives used in the preparation of general formulations may be contained. Examples of such additives include excipients, binders, acidulants, foaming agents, artificial sweeteners, flavors, lubricants, coloring agents, stabilizers, pH adjusters, surfactants and the like.

  When a liquid dosage form is used, the composition of the present invention can be prepared by a conventional method in the presence of a pH adjusting agent, a buffer, a dissolving agent, a suspending agent, etc., a tonicity agent, a stabilizing agent, a preservative, etc. It can be formulated.

  The form of the external preparation for skin is not particularly limited. For example, emulsions, creams, lotions, packs, dispersions, detergents, makeup cosmetics, cosmetics such as scalp and hair products, ointments, creams, external use It can be used as a drug such as a liquid drug. In addition to the above components, components usually used for external skin preparations such as cosmetics and pharmaceuticals, for example, skin lightening agents, moisturizers, various skin nutritional components, ultraviolet light absorbers, antioxidants, oil components, surfactants, thickeners Alcohols, coloring agents, water, preservatives, perfumes and the like can be appropriately blended as required.

  The composition of the present invention is prepared as functional food, health food, food for specified health use, health functional food such as nutritive function food, food for special use (for example, food for sickness), health food supplement, supplement etc. It is also good. As a supplement, for example, tablets, rounds, capsules (including hard capsules, soft capsules, and microcapsules), powders, granules, fine particles, troches, and liquids, together with various additives used for the production of common supplements It can be in the form of syrup (including syrup, milk, suspension) and the like.

  The composition of the present invention can be formulated into food. There are no particular limitations on the foods that can be incorporated, but, for example, confectionery such as rice cake, gummi, chewing gum, etc .; Ice cream candy, sherbet, gelato frozen desserts; bakery foods such as donuts, cakes, bread, french bread, croissants, etc .; noodles such as udon, buckwheat noodles, Chinese noodles, Chinese noodles; cooked rice such as white rice, red rice, pilaf; curry, stew , Sauces such as dressings; Braised products such as ham, sausages, kamaboko, chikuwa, fish meat sausages; Various vegetables such as tempuras, croquettes and hamburgs;

  EXAMPLES The present invention will be illustrated specifically by the following examples, but the present invention is not limited to the following examples.

Test Example 1 [Method of producing oleanan triterpene derived from olive fruit]
Olive oil was removed by a known pressing method such as pressing to obtain an olive fruit pressed rice cake. After adding 250g of 90% hydrous ethanol (v / v) to 100g of this olive fruit oil cake and heating for 2 hours while stirring at 70 ° C, the liquid portion obtained by solid-liquid separation is evaporated under reduced pressure with an evaporator etc. A concentrated solution was obtained. This concentrated solution is dispersed in water and subjected to column chromatography using synthetic adsorption resin Amberlite XAD4 (manufactured by Organo) as a carrier, and the triterpene height is increased by stepwise elution from 50% water-containing ethanol to 100% ethanol in 10% steps. A fraction containing the product was obtained and evaporated under reduced pressure to obtain a liquid olive fruit-derived oleanane type triterpene. The triterpene-containing solution contained masulinic acid and oleanolic acid in a ratio of 10: 4 (FIG. 1). Peak identification was performed by comparison of retention time with standard products and LC / MS analysis. The quantitative value was calculated based on the peak area of the standard product.
After adding and mixing (gamma) -cyclodextrin so that a mass phosphoric acid content might be 10 weight%, it disperse | distributed to water, it spray-dried and made it the olive fruit origin oleanane type triterpene agent of powder.

Test Example 2 [Preparation of Triterpene-Containing Supplement]
The raw materials were mixed according to the formulation shown in Table 1 and filled in a gelatin hard capsule so that the content of the mixture was 250 mg. The maslinic acid content per capsule obtained in this test example is 17 mg, and the oleanolic acid content is 6.8 mg.

Experiment 3 [Evaluation in patients with mild osteoarthritis of the knee]
(1) Criteria of the method and effect of clinical test and selection of subjects The subjects were 20 patients who were diagnosed with mild knee osteoarthritis by interview and did not need treatment. The average age is 58.1 years, 1 male and 19 female.
(2) How to take the triterpene-containing supplement 12 people were randomly selected, and the triterpene-containing supplement obtained in Test Example 2 was once daily once breakfast 3 capsules (51 mg / day as maslinic acid) daily for 12 weeks I took it. The remaining eight patients similarly took the capsule of the comparative example.
(3) Clinical marker test Blood was collected before and after taking the capsule, and cartilage synthesis marker (PIIANP), cartilage degradation marker (C2C) and inflammation marker (hsCRP) were measured. The results are shown in Tables 2, 3 and 4 below.
The cartilage synthesis marker was measured using PIIANP ELISA kit (manufactured by Millipore). Cartilage degradation markers were measured using Collagen Type II Cleavage ELISA (manufactured by IBEX). Inflammation markers were measured using N-latex CRPII (manufactured by Siemens Health Care Diagnostics).

*: ± is standard error.
In the example, the blood PIIANP was raised by the dose, but in the comparative example it was slightly decreased (Table 2). From this fact, triterpenes derived from olive fruits were found to have an effect of promoting cartilage synthesis.

*: ± is standard error.
In the example, C2C in the blood was increased by taking it, but in the comparative example, it was only slightly increased (Table 3). From this, triterpenes derived from olive fruits were found to have the effect of promoting the degradation of the surface layer of cartilage.

*: ± is standard error.
In the example, blood hsCRP decreased by taking, but increased in the comparative example (Table 4). From this fact, olive fruit-derived triterpenes were found to be effective in suppressing inflammation.
Summarizing the results in Tables 2 to 4 above, it can be concluded that triterpenes derived from olive fruits have an effect of promoting cartilage synthesis and degradation of the cartilage surface layer, and conclude that metabolism is carried out in the cartilage part It was shown that the ability to promote cartilage collagen metabolism is effective.

(4) Evaluation of VAS VAS (Visual Analog Scale) is an evaluation method of pain widely used in the medical field, and the 10 cm straight ends are "painless" and "the most severe pain ever experienced"", And is a way to indicate where the current pain is on 10 cm. It was carried out before the start of dosing and after the end of dosing. The results are shown in Table 5 below.

*: ± is standard error.
Although the placebo effect occurred in the comparative example, the improvement effect of VAS was recognized more significantly in the example.
(5) Evaluation of QOL (Quality of Life) Evaluation of QOL was carried out before and after taking the drug according to the “SF-8TM Japanese edition manual” issued by the NPO Health and Medical Evaluation Research Organization. The results are shown in Table 6 below.
*: ± is standard error.
The placebo effect was produced in the comparative example, but the QOL improvement effect was more remarkable in the example.

From the above results, it was found that olive fruit-derived oleanan triterpenes promote cartilage synthesis and cartilage degradation and promote cartilage metabolism. In addition, it had anti-inflammatory and pain-relieving effects, and tended to improve VAS and QOL of subjects.
With regard to other safety, as a result of blood test, urine test, physical measurement and medical examination, no medically problematic fluctuation due to intake of oleanane type triterpene derived from olive fruit was found.

Claims (3)

A composition for promoting cartilage collagen metabolism, comprising a triterpene or a pharmacologically acceptable salt thereof as an active ingredient,
The triterpene is an oleanane-type triterpene derived from olive fruit oil persimmon containing maslinic acid and oleanolic acid,
The composition as described above, except that the triterpene contains maslinic acid and oleanolic acid in a ratio of 10: 4 and the composition contains curcuminoid species. A composition for promoting cartilage collagen metabolism, comprising a triterpene or a pharmacologically acceptable salt thereof as an active ingredient,
The triterpene consists of maslinic acid and oleanolic acid,
The triterpene contains maslinic acid and oleanolic acid in a ratio of 10: 4 ,
A composition as above except where the composition comprises curcuminoid species. The composition according to claim 1 or 2, which is orally administered.