JP2004083434A - Accelerator for collagen synthesis - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a novel highly safe accelerator for collagen synthesis. <P>SOLUTION: The novel accelerator for collagen synthesis can be provided as the result of a finding that a glucosamine, a glucosamine derivative, or salts thereof have an accelerating effect for collagen synthesis. The accelerator for collagen synthesis is highly effective, to alleviate decrease in elasticity of the skin, to ameliorate wrinkles or sags of the skin, and to prevent and/or treat joint disorders. <P>COPYRIGHT: (C)2004,JPO

Description

【００１７】
表１に示す結果より、塩酸グルコサミンは、濃度依存的に皮膚繊維芽細胞においてコラーゲン合成量を促進し、Ｉ型コラーゲンの合成量を２．５倍に増加することが確認された。
【００１８】
試験例２　へアレスマウスを用いた抗シワ試験
試験例２−１
表３に示す実施例１〜３について、紫外線によるシワ発生に対する予防効果を塩酸グルコサミンの塗布試験により検討した。５週齡雄性へアレスマウスは８匹を１群とした４群に対し、３週間にわたり、第１週目に９０ｍＪ／ｃｍ^２、第２週目に１２０ｍＪ／ｃｍ^２、第３週目に１５０ｍＪ／ｃｍ^２のＵＶＢ紫外線を１週間あたり３回照射した。また紫外線を照射している３週間、各群のヘアレスマウスには、実施例１〜３又は比較例１の試験溶液を５０μｌずつ１日３回背中に塗布した。そして、第１回目の紫外線照射から２４日後に、目視にてシワを７段階にスコア化（表２）し、シワ発生の予防効果を評価した。結果は表３に示す。
【００１９】
試験例２−２
表３に示す実施例４〜６について、紫外線によるシワ発生に対する予防効果を塩酸グルコサミンの経口投与により検討した。試験例２−１と同様に５週齡雄性へアレスマウスは８匹を１群とした４群に対し、３週間にわたりＵＶＢ紫外線を照射した。また、紫外線を照射している３週間の間、各群のヘアレスマウスには、実施例４〜６又は比較例３の試験溶液を３００μｌずつ１日３回経口投与し、実施例４では塩酸グルコサミンが５００ｍｇ／ｋｇｗ／ｄａｙ、実施例５では塩酸グルコサミンが１０００ｍｇ／ｋｇｗ／ｄａｙ、実施例６では塩酸グルコサミンが１５００ｍｇ／ｋｇｗ／ｄａｙとなるように投与した。そして、第１回目の紫外線照射から２４日後に、目視にてシワを７段階にスコア化（表２）し、シワ発生の予防効果を評価した。結果は表３に示す。
【００２０】
【表２】

【００２１】
【表３】

【００２２】
表３に示す結果より、塩酸グルコサミンを含有する実施例は、濃度依存的に紫外線曝露によるシワの予防効果があることが確認された。塩酸グルコサミンは、皮膚のシワの形成を抑制する優れた抗シワ効果があることがわかった。
【００２３】
実施例７　下記の他の成分を日本薬局方製剤総則「錠剤」に準じて、混合し打錠して円形錠剤（１錠５００ｍｇ）を得た。

【００２４】

【００２５】

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a collagen synthesis promoter containing glucosamine, a derivative thereof, or a salt thereof.
[0002]
[Prior art]
Collagen is a major protein that makes up the connective tissue of animals, and in particular, it accounts for nearly 30% of the total protein in the human body. Since the main function of collagen is to form the skeletal structure of living tissue, it is widely distributed in skin, cartilage tissue, cornea, heart, liver, and the like as a main component of the skeletal structure of animal tissue. Collagen specifically acts on the adhesion of various cells, differentiation and proliferation of cells, and also has a role as a solid phase regulator of cell function, so collagen degradation causes corneal disorders such as corneal ulcers It may cause various diseases such as rheumatism, arthritis, osteoarthritis, osteoarthritis and other joint disorders, and various inflammatory diseases.
[0003]
In the skin dermis extracellular matrix, collagen fibers maintain a tissue morphology by forming a mesh-like bundle. Collagen fibers mature and proliferate to form thick, linear fiber bundles as cross-linking progresses, providing moderate tension in young skin. However, as the activity of fibroblasts decreases in aged or photoaged skin, collagen fibers in the extracellular matrix of the dermis are significantly reduced, and abnormal aging crosslinks are formed, resulting in rigidity and The elasticity of the elasticity is lost. As a result, wrinkles and lumps are formed on the skin. Changes in the fiber bundle structure of the hairless mouse due to photoaging are examined in detail (Fragrance Journal, 4, p36-37, 1998). It has been shown that the collagen fiber bundle structure collapses and skin elasticity decreases. It is also known that collagen has an excellent water retention function.
[0004]
Various collagen synthesis promoting substances have been found to improve such a condition due to collagen reduction, for example, retinoic acid (R. Marks et al, British Journal of Dermatology, 122, 91, 1990), Three kinds of amino acids consisting of glycine, proline, and alanine (JP-A-7-194375); TGF-β, ascorbic acid and derivatives thereof are known. However, they have not been fully satisfactory in terms of safety and effect.
[0005]
[Problems to be solved by the invention]
An object of the present invention is to provide a novel collagen synthesis promoter having high safety.
[0006]
[Means for Solving the Problems]
The present inventors have conducted intensive studies to solve the above problems, and as a result, have found that glucosamine, a glucosamine derivative or a salt thereof has a collagen synthesis promoting effect, and have completed the present invention. Glucosamine, a glucosamine derivative or a salt thereof has a glycosaminoglycan secretion effect using a culture solution of mouse fibroblasts (K. Karzel, et. Al., Pharmacology, 5, p337-, 1971). However, the relationship with collagen synthesis has not been reported yet. That is, the present invention relates to a collagen synthesis promoter containing at least one compound selected from glucosamine, a glucosamine derivative, or a pharmacologically acceptable salt thereof. Further, the present invention relates to a collagen synthesis promoter comprising as an active ingredient at least one compound selected from glucosamine, a derivative of glucosamine or a pharmaceutically acceptable salt thereof.
[0007]
BEST MODE FOR CARRYING OUT THE INVENTION
Glucosamine, a glucosamine derivative or a pharmaceutically acceptable salt thereof in the present invention is preferably a natural product purified from animals, plants, microbial cells, culture solutions, and the like, but is chemically synthesized by a known method. The obtained synthetic product may be used, or a commercially available product may be used. In the present invention, as the glucosamine derivative, a compound that can be converted into glucosamine after being applied in a living body is preferable, and examples thereof include a derivative such as N-acetylglucosamine. The glucosamine derivative is not particularly limited as long as the effect of the present invention is exerted.
[0008]
As the salt of the glucosamine or glucosamine derivative of the present invention, a pharmacologically (pharmaceutically) or physiologically acceptable salt can be used. Pharmaceutically or physiologically acceptable salts include, for example, organic acid salts (eg, lactate, acetate, butyric acid, trifluoroacetate, fumarate, maleate, tartrate, citrate, succinate) Acid salt, malonate, methanesulfonate, toluenesulfonate, tosylate, palmitic acid, stearic acid, etc.), inorganic acid salts (for example, hydrochloride, sulfate, nitrate, hydrobromide, phosphorus) Acid salts). Preferably, glucosamine hydrochloride and glucosamine sulfate are used. Particularly preferred is glucosamine hydrochloride.
These glucosamines, derivatives thereof or salts thereof may be used alone or in combination of two or more.
[0009]
In the elastase inhibitor of the present invention, the amount of glucosamine, a glucosamine derivative or a salt thereof used varies depending on the type and dosage form of glucosamine. A usage amount of 0.01 to 3000 mg, preferably 0.1 to 2000 mg, particularly preferably 0.1 to 2000 mg can be applied once or in 2 to 4 times. When an elastase inhibitor that is not applicable to mammals including humans, such as a research reagent, is used, the amount used can be appropriately set according to the dosage form and the like.
[0010]
The content of glucosamine, a glucosamine derivative or a salt thereof in the collagen synthesis promoter of the present invention varies depending on the type, dosage form and the like of glucosamine, but is usually 0.0001 to 70.0% by weight, preferably 0.1% by weight. The range is 001 to 50% by weight, more preferably 0.001 to 20% by weight, and particularly preferably 0.01 to 10% by weight.
[0011]
The collagen synthesis promoter of the present invention can be used in a quantitative and qualitative range that does not impair the effects of the present invention, in addition to glucosamine, a glucosamine derivative or a salt thereof, and commonly used solvents, bases, and various additives (for example, surfactants). Activators, emulsifiers, oils, stabilizers, thickeners, preservatives, binders, lubricants, coloring agents, dispersants, pH adjusters, humectants, ultraviolet absorbers, vitamins, chelating agents, amino acids , Fragrances, etc.), as well as solid preparations such as tablets, granules, powders, fine granules and hard capsules, liquid preparations such as syrups and soft capsules, semi-solid preparations such as ointments and emulsions In the case of preparations such as preparations and cosmetics, various desired forms such as foundations, lipsticks, lotions, serums, emulsions, creams, lotions, oils, gels and packs can be prepared. These can be produced by a conventional method using commonly used additives and solvents.
[0012]
The collagen synthesis promoter of the present invention can be used in various fields such as pharmaceuticals, quasi-drugs, foods, and cosmetics. Specifically, it is useful as a therapeutic or prophylactic agent for diseases caused by a decrease in the amount of collagen in mammals including humans, or as a research reagent for elucidating a physiological state related to collagen. Specifically, it is used as a therapeutic and / or prophylactic agent for joint diseases such as rheumatoid arthritis and osteoarthritis, and for preventing and / or improving skin wrinkles and lumps due to UV exposure and aging, and skin elasticity. And anti-aging or external skin preparations for beautiful skin for preventing or improving the reduction of firmness.
[0013]
【The invention's effect】
Glucosamine, a glucosamine derivative or a salt thereof has an action of promoting collagen synthesis. Therefore, it prevents the deterioration of elasticity and firmness, which are skin aging phenomena caused by the effects of aging, ultraviolet rays, etc., enhances the moisturizing effect, recovers or maintains the firmness of the skin, and keeps the skin condition youthful be able to. It is also useful for preventing or improving wrinkles and tarmi. Further, it is also effective for treating various diseases such as joint diseases.
[0014]
【Example】
Hereinafter, the present invention will be described in more detail based on examples and test examples, but the present invention is not limited to these examples and the like. In each of the following formulations,% means weight (W / W)% unless otherwise specified.
[0015]
Test Example 1 Normal human skin fibroblasts (NHDF; manufactured by Kurabo Industries) were seeded at 2 × 10 ⁴ cells / well on a 24-well culture plate, and the skin fibroblasts were used as a culture solution. Using a growth medium (manufactured by Kurabo Industries), the cells were cultured at 37 ° C. under 5% CO ₂ . After culturing for 72 hours, the culture medium was removed from each well, and the above culture medium containing glucosamine hydrochloride at a concentration of 0.0002, 0.002, and 0.02% or a culture medium containing no glucosamine hydrochloride as a control was added to each well. 1 ml was added to the wells, and the cells were further cultured for 72 hours. After completion of the culture, the amount of type I procollagen in the culture supernatant was measured by ELISA (Procollagen type I C-peptide EIA Kit; manufactured by Takara Shuzo). Since type I procollagen is a precursor of type I collagen, the amount of type I procollagen was measured to be used as a biochemical index of the amount of collagen synthesis. Table 1 shows the results. Each value is a ratio to the amount of type I procollagen synthesized in the control to which glucosamine hydrochloride was not added.
[0016]
[Table 1]

[0017]
From the results shown in Table 1, it was confirmed that glucosamine hydrochloride promoted the amount of collagen synthesis in skin fibroblasts in a concentration-dependent manner and increased the amount of type I collagen synthesis 2.5-fold.
[0018]
Test Example 2 Anti-wrinkle test using hairless mouse Test Example 2-1
With respect to Examples 1 to 3 shown in Table 3, the preventive effect against the generation of wrinkles due to ultraviolet rays was examined by an application test of glucosamine hydrochloride. To 5 weeks齡雄property to hairless mice group 4 was Eight and one group, 150 mJ for 3 weeks, 90 mJ / cm ² in the first week, the second week 120 mJ / cm ^2, the third week / Cm ² of UVB ultraviolet light was irradiated three times per week. In addition, for 3 weeks while irradiating ultraviolet rays, 50 μl of the test solution of Examples 1 to 3 or Comparative Example 1 was applied to the back of the hairless mice of each group three times a day. Twenty-four days after the first ultraviolet irradiation, wrinkles were visually scored into seven levels (Table 2) to evaluate the effect of preventing wrinkles from occurring. The results are shown in Table 3.
[0019]
Test Example 2-2
With respect to Examples 4 to 6 shown in Table 3, the preventive effect against wrinkles caused by ultraviolet rays was examined by oral administration of glucosamine hydrochloride. Similar to Test Example 2-1, five-week-old male hairless mice were irradiated with UVB ultraviolet rays for three weeks to four groups of eight mice. In addition, during the three weeks of irradiation with ultraviolet light, the hairless mice in each group were orally administered 300 μl of the test solution of Examples 4 to 6 or Comparative Example 3 three times a day. In Example 4, glucosamine hydrochloride was used. Was administered at 500 mg / kgw / day, in Example 5, glucosamine hydrochloride was administered at 1000 mg / kgw / day, and in Example 6, glucosamine hydrochloride was administered at 1500 mg / kgw / day. Twenty-four days after the first ultraviolet irradiation, wrinkles were visually scored into seven levels (Table 2) to evaluate the effect of preventing wrinkles from occurring. The results are shown in Table 3.
[0020]
[Table 2]

[0021]
[Table 3]

[0022]
From the results shown in Table 3, it was confirmed that Examples containing glucosamine hydrochloride had a concentration-dependent effect of preventing wrinkles due to exposure to ultraviolet light. Glucosamine hydrochloride was found to have an excellent anti-wrinkle effect for suppressing the formation of skin wrinkles.
[0023]
Example 7 The following other components were mixed and compressed according to the Japanese Pharmacopoeia General Rules for Preparations “Tablets” to give circular tablets (500 mg per tablet).

[0024]

[0025]

Claims (1)

A collagen synthesis promoter containing at least one selected from glucosamine, a glucosamine derivative or a pharmaceutically acceptable salt thereof.