JP6988016B1 - Granzyme B inhibitor - Google Patents

Granzyme B inhibitor Download PDF

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JP6988016B1
JP6988016B1 JP2021072466A JP2021072466A JP6988016B1 JP 6988016 B1 JP6988016 B1 JP 6988016B1 JP 2021072466 A JP2021072466 A JP 2021072466A JP 2021072466 A JP2021072466 A JP 2021072466A JP 6988016 B1 JP6988016 B1 JP 6988016B1
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紘子 山嵜
美怜 山野
真穂 横田
大嗣 亀井
良樹 伊藤
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Sato Pharmaceutical Co Ltd
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Abstract

【課題】効果の高いグランザイムＢ抑制剤を提供する。【解決手段】ゲンノショウコエキスを有効成分として含有する。【選択図】なしPROBLEM TO BE SOLVED: To provide a highly effective Granzyme B inhibitor. SOLUTION: Geranium thunbergii extract is contained as an active ingredient. [Selection diagram] None

Description

本発明は、ゲンノショウコエキスを有効成分として含むグランザイムＢ抑制剤に関する。 The present invention relates to a Granzyme B inhibitor containing Geranium thunbergii extract as an active ingredient.

グランザイムＢ阻害剤は、免疫誘導効果、自己免疫性・慢性の炎症性疾患への効果が示唆されている（特許文献１〜２）。また、皮膚への適用においては、火傷、創傷治癒の促進効果、皮膚老化に対する外観改善効果が示されている（特許文献３〜５）。
グランザイムＢ活性阻害剤として、既知の低分子ペプチド等が知られている（特許文献１〜３、非特許文献１〜２）。しかし、原料の入手が容易で安価な天然由来物質のグランザイムＢ活性抑制剤に関する報告はない。
また、グランザイムＢは、分化誘導性のサイトカインにより発現誘導され、アポトーシスに関与する（非特許文献３）。皮膚においては通常状態における発現量は低く、自己免疫性・慢性炎症性皮膚状態で発現量が増加すること、紫外線照射によっても発現誘導される（非特許文献４〜６）。しかしながら、グランザイムＢ発現を抑制する素材に関する報告は皆無である。 Granzyme B inhibitors have been suggested to have immunostimulatory effects and effects on autoimmune and chronic inflammatory diseases (Patent Documents 1 and 2). In addition, when applied to the skin, it has been shown to have an effect of promoting burns and wound healing, and an effect of improving the appearance of skin aging (Patent Documents 3 to 5).
Known small molecule peptides and the like are known as granzyme B activity inhibitors (Patent Documents 1 to 3 and Non-Patent Documents 1 to 2). However, there are no reports on granzyme B activity inhibitors, which are naturally derived substances that are easily available as raw materials and are inexpensive.
In addition, Granzyme B is induced to be expressed by differentiation-inducing cytokines and is involved in apoptosis (Non-Patent Document 3). In the skin, the expression level is low in the normal state, the expression level is increased in the autoimmune / chronic inflammatory skin state, and the expression level is also induced by ultraviolet irradiation (Non-Patent Documents 4 to 6). However, there are no reports on materials that suppress the expression of Granzyme B.

国際公開第２００３／０６５９８７号International Publication No. 2003/06598 米国特許出願公開第２００３／０１４８５１１号明細書U.S. Patent Application Publication No. 2003/01485511 国際公開第２０１７／１３２７７１号International Publication No. 2017/132771 特許第６１３４２６８号公報Japanese Patent No. 6134268 特許第５７４６８１３号公報Japanese Patent No. 5746813

Bird et. al., Mol. Cell. Biol., 1998, 18, 6387-6398Bird et. Al., Mol. Cell. Biol., 1998, 18, 6387-6398 Kam et. al., Biochim. Biophys. Acta., 2000, 1477(1-2), 307-23Kam et. Al., Biochim. Biophys. Acta., 2000, 1477 (1-2), 307-23 Tamang et. al., Cytokine, 2006 November, 36(3-4), 148-159Tamang et. Al., Cytokine, 2006 November, 36 (3-4), 148-159 Turner et.al., J. Invest. Dermatol., 2021, Jan, 141(1), 36-47Turner et.al., J. Invest. Dermatol., 2021, Jan, 141 (1), 36-47 Hiebert et.al., Exp. Gerontol., 2011 Jun, 46(6),489-99Hiebert et.al., Exp. Gerontol., 2011 Jun, 46 (6), 489-99 Parkinson et.al., Aging Cell, 2015 Feb, 14(1), 67-77Parkinson et.al., Aging Cell, 2015 Feb, 14 (1), 67-77

グランザイムＢの発現上昇を抑え、かつグランザイムＢ活性阻害作用も有する素材は、グランザイムＢに起因する皮膚への増悪作用を効率よく抑制することが示唆される。
したがって、グランザイムＢ発現抑制作用及び活性阻害作用の両作用を有する天然由来物質は、より応用範囲が広いグランザイムＢ作用の抑制剤としての利用が期待される。 It is suggested that the material that suppresses the increase in the expression of granzyme B and also has the action of inhibiting the activity of granzyme B efficiently suppresses the exacerbating action on the skin caused by granzyme B.
Therefore, naturally derived substances having both an action of suppressing the expression of granzyme B and an action of inhibiting the activity are expected to be used as an inhibitor of the action of granzyme B, which has a wider range of applications.

本発明者は、広範囲の植物エキスのうち、ゲンノショウコエキスがグランザイムＢ発現抑制作用及び活性阻害作用の両作用を有することを見出し、本発明を完成するに至った。また、本発明者はさらに、複数の植物エキス添加によりグランザイムＢ活性阻害作用が増強することを示した。すなわち、本発明は下記の〔１〕〜〔２３〕に関するものである。
〔１〕ゲンノショウコエキスを有効成分として含有する、グランザイムＢ抑制剤。
〔２〕ゲンノショウコエキスを１〜１００μｇ／ｍＬ含有する、前記〔１〕に記載のグランザイムＢ抑制剤。
〔３〕ゲンノショウコエキスを１．８〜５５μｇ／ｍＬ含有する、前記〔２〕に記載のグランザイムＢ抑制剤。
〔４〕ゲンノショウコエキスを１８〜５５μｇ／ｍＬ含有する、前記〔３〕に記載のグランザイムＢ抑制剤。
〔５〕グランザイムＢ抑制作用を有する１種以上の植物エキスをさらに含有する、前記〔１〕から〔４〕のいずれか一項に記載のグランザイムＢ抑制剤。
〔６〕前記植物エキス及び前記ゲンノショウコエキスを１：１〜１：４の質量比で含有する、前記〔５〕に記載のグランザイムＢ抑制剤。
〔７〕前記植物エキスが、ザクロ果皮エキス、イエローヒマラヤンラズベリー根エキス、ユーカリエキス、ワレモコウエキス、イロハモミジ葉エキス、シモツケソウエキス、ハマメリスエキス、バラエキス、アボガドエキス、テンチャエキス、メマツヨイグサ抽出液、ウーロン茶エキス、カシア樹皮エキス、ユキノシタエキス、メリッサエキス、トルメンチラエキス、ダマスクバラ花エキス、ヨモギエキス、ヒキオコシエキス、コンフリーエキス、オトギリソウエキス、セイヨウハッカエキス、ブドウ葉エキス、シナノキエキス、エイジツエキス、ビワ葉エキス、海藻エキス、シャクヤクエキス、ドクダミエキス、サンザシエキス、シソエキス、ラベンダーエキス、ノバラエキス、アセンヤクエキス、サガラメエキス、ボタンエキス、アスパラサスリネアリスエキス、カンゾウ抽出末及びクララエキスからなる群から選択される、前記〔５〕又は〔６〕に記載のグランザイムＢ抑制剤。
〔８〕前記植物エキスの少なくとも１種が、オトギリソウエキス及びテンチャエキスからなる群から選択される、前記〔７〕に記載のグランザイムＢ抑制剤。
〔９〕グランザイムＢ抑制が、グランザイムＢ遺伝子の発現抑制である、前記〔１〕から〔８〕のいずれか一項に記載のグランザイムＢ抑制剤。
〔１０〕グランザイムＢ抑制が、グランザイムＢの活性阻害である、前記〔１〕から〔８〕のいずれか一項に記載のグランザイムＢ抑制剤。
〔１１〕グランザイムＢ抑制が、グランザイムＢ遺伝子の発現抑制及びグランザイムＢの活性阻害である、前記〔１〕から〔８〕のいずれか一項に記載のグランザイムＢ抑制剤。
〔１２〕前記〔１〕から〔１１〕のいずれか一項に記載のグランザイムＢ抑制剤のみからなる、化粧料組成物。
〔１３〕ゲンノショウコエキスを有効成分として含有する、コラーゲン分解抑制剤。
〔１４〕ゲンノショウコエキスを１〜１００μｇ／ｍＬ含有する、前記〔１３〕に記載のコラーゲン分解抑制剤。
〔１５〕ゲンノショウコエキスを１．８〜５５μｇ／ｍＬ含有する、前記〔１４〕に記載のコラーゲン分解抑制剤。
〔１６〕ゲンノショウコエキスを１８〜５５μｇ／ｍＬ含有する、前記〔１５〕に記載のコラーゲン分解抑制剤。
〔１７〕コラーゲン分解抑制作用を有する１種以上の植物エキスをさらに含有する、前記〔１３〕から〔１６〕のいずれか一項に記載のコラーゲン分解抑制剤。
〔１８〕前記植物エキス及び前記ゲンノショウコエキスを１：１〜１：４の質量比で含有する、前記〔１７〕に記載のコラーゲン分解抑制剤。
〔１９〕前記植物エキスが、ザクロ果皮エキス、イエローヒマラヤンラズベリー根エキス、ユーカリエキス、ワレモコウエキス、イロハモミジ葉エキス、シモツケソウエキス、ハマメリスエキス、バラエキス、アボガドエキス、テンチャエキス、メマツヨイグサ抽出液、ウーロン茶エキス、カシア樹皮エキス、ユキノシタエキス、メリッサエキス、トルメンチラエキス、ダマスクバラ花エキス、ヨモギエキス、ヒキオコシエキス、コンフリーエキス、オトギリソウエキス、セイヨウハッカエキス、ブドウ葉エキス、シナノキエキス、エイジツエキス、ビワ葉エキス、海藻エキス、シャクヤクエキス、ドクダミエキス、サンザシエキス、シソエキス、ラベンダーエキス、ノバラエキス、アセンヤクエキス、サガラメエキス、ボタンエキス、アスパラサスリネアリスエキス、カンゾウ抽出末及びクララエキスからなる群から選択される、前記〔１７〕又は〔１８〕に記載のコラーゲン分解抑制剤。
〔２０〕前記植物エキスの少なくとも１種が、オトギリソウエキス及びテンチャエキスからなる群から選択される、前記〔１９〕に記載のコラーゲン分解抑制剤。
〔２１〕コラーゲン分解抑制が、グランザイムＢ遺伝子の発現抑制である、前記〔１３〕から〔２０〕のいずれか一項に記載のコラーゲン分解抑制剤。
〔２２〕コラーゲン分解抑制が、グランザイムＢの活性阻害である、前記〔１３〕から〔２０〕のいずれか一項に記載のコラーゲン分解抑制剤。
〔２３〕コラーゲン分解抑制が、グランザイムＢ遺伝子の発現抑制及びグランザイムＢの活性阻害である、前記〔１３〕から〔２０〕のいずれか一項に記載のコラーゲン分解抑制剤。 The present inventor has found that Geranium thunbergii extract has both an action of suppressing the expression of granzyme B and an action of inhibiting the activity among a wide range of plant extracts, and has completed the present invention. In addition, the present inventor has further shown that the addition of a plurality of plant extracts enhances the action of inhibiting granzyme B activity. That is, the present invention relates to the following [1] to [23].
[1] A Granzyme B inhibitor containing Geranium thunbergii extract as an active ingredient.
[2] The granzyme B inhibitor according to the above [1], which contains 1 to 100 μg / mL of Geranium thunbergii extract.
[3] The granzyme B inhibitor according to the above [2], which contains 1.8 to 55 μg / mL of Geranium thunbergii extract.
[4] The granzyme B inhibitor according to the above [3], which contains 18 to 55 μg / mL of Geranium thunbergii extract.
[5] The granzyme B inhibitor according to any one of the above [1] to [4], which further contains one or more plant extracts having a granzyme B inhibitory action.
[6] The granzyme B inhibitor according to the above [5], which contains the plant extract and the geranium thunbergii extract in a mass ratio of 1: 1 to 1: 4.
[7] The plant extracts include pomegranate peel extract, yellow Himalayan raspberry root extract, eucalyptus extract, crackle extract, Irohamomiji leaf extract, Shimotsukesou extract, Hamamelis extract, rose extract, avocado extract, tencha extract, mematsuyoigusa extract, oolong tea extract, and cassia. Bark extract, Yukinoshita extract, Melissa extract, Tormentilla extract, Damask rose flower extract, Yomogi extract, Hikiokoshi extract, Confree extract, Otogirisou extract, Seiyouhakka extract, Grape leaf extract, Shinanoki extract, Agetsu extract, Biwa leaf extract, The above-mentioned [ 5] or the Granzyme B inhibitor according to [6].
[8] The Granzyme B inhibitor according to the above [7], wherein at least one of the plant extracts is selected from the group consisting of Hypericum erectum extract and Tencha extract.
[9] The granzyme B inhibitor according to any one of [1] to [8] above, wherein the granzyme B inhibition is the inhibition of the expression of the granzyme B gene.
[10] The Granzyme B inhibitor according to any one of [1] to [8] above, wherein the inhibition of Granzyme B is the inhibition of the activity of Granzyme B.
[11] The Granzyme B inhibitor according to any one of [1] to [8] above, wherein the suppression of Granzyme B is the suppression of the expression of the Granzyme B gene and the inhibition of the activity of Granzyme B.
[12] A cosmetic composition comprising only the Granzyme B inhibitor according to any one of the above [1] to [11].
[13] A collagen decomposition inhibitor containing Geranium thunbergii extract as an active ingredient.
[14] The collagen decomposition inhibitor according to the above [13], which contains 1 to 100 μg / mL of Geranium thunbergii extract.
[15] The collagen decomposition inhibitor according to the above [14], which contains 1.8 to 55 μg / mL of Geranium thunbergii extract.
[16] The collagen decomposition inhibitor according to the above [15], which contains 18 to 55 μg / mL of Geranium thunbergii extract.
[17] The collagen decomposition inhibitor according to any one of [13] to [16] above, further containing one or more plant extracts having a collagen decomposition inhibitory effect.
[18] The collagen decomposition inhibitor according to the above [17], which contains the plant extract and the geranium thunbergii extract in a mass ratio of 1: 1 to 1: 4.
[19] The plant extracts include pomegranate peel extract, yellow Himalayan raspberry root extract, eucalyptus extract, crackle extract, Irohamomiji leaf extract, Shimotsukesou extract, Hamamelis extract, rose extract, avocado extract, tencha extract, mematsuyoigusa extract, oolong tea extract, and cassia. Bark extract, Yukinoshita extract, Melissa extract, Tormentilla extract, Damask rose flower extract, Yomogi extract, Hikiokoshi extract, Confree extract, Otogirisou extract, Seiyouhakka extract, Grape leaf extract, Shinanoki extract, Agetsu extract, Biwa leaf extract, The above-mentioned [ 17] or the collagen decomposition inhibitor according to [18].
[20] The collagen decomposition inhibitor according to the above [19], wherein at least one of the plant extracts is selected from the group consisting of Hypericum erectum extract and Tencha extract.
[21] The collagen degradation inhibitor according to any one of [13] to [20] above, wherein the inhibition of collagen degradation is the inhibition of the expression of the Granzyme B gene.
[22] The collagen degradation inhibitor according to any one of [13] to [20] above, wherein the inhibition of collagen degradation is an inhibition of the activity of Granzyme B.
[23] The collagen degradation inhibitor according to any one of [13] to [20] above, wherein the inhibition of collagen degradation is the inhibition of the expression of the Granzyme B gene and the inhibition of the activity of Granzyme B.

後述の実施例で示されるように、本発明に従うゲンノショウコエキスはグランザイムＢ発現抑制作用及び活性阻害作用を有するので、グランザイムＢ抑制剤として有用である。 As shown in Examples described later, Geranium thunbergii extract according to the present invention has a granzyme B expression inhibitory action and an activity inhibitory action, and is therefore useful as a granzyme B inhibitor.

図１は、ゲンノショウコエキスのグランザイムＢ遺伝子の発現抑制作用を示す実験結果である。FIG. 1 shows the experimental results showing the inhibitory effect of Geranium thunbergii extract on the expression of the Granzyme B gene. 図２Ａは、ゲンノショウコエキスのグランザイムＢの活性阻害作用を示す実験結果である。FIG. 2A is an experimental result showing the activity inhibitory effect of Granzyme B of Geranium thunbergii extract. 図２Ｂは、ゲンノショウコエキスのグランザイムＢの活性阻害作用を示す実験結果である。FIG. 2B is an experimental result showing the activity inhibitory effect of Granzyme B of Geranium thunbergii extract. 図３は、グランザイムＢの活性阻害作用を有する植物エキスを示す。FIG. 3 shows a plant extract having an activity inhibitory effect on Granzyme B. 図４Ａは、オトギリソウエキスのグランザイムＢの活性阻害作用を示した図である。FIG. 4A is a diagram showing the activity inhibitory effect of Granzyme B of Hypericum erectum extract. 図４Ｂは、テンチャエキスのグランザイムＢの活性阻害作用を示した図である。FIG. 4B is a diagram showing the activity inhibitory effect of Granzyme B of Tencha extract. 図５Ａは、ゲンノショウコエキスとオトギリソウエキスの、グランザイムＢ抑制によるデコリン分解抑制の相乗効果を示した図である。FIG. 5A is a diagram showing the synergistic effect of Geranium thunbergii extract and Hypericum erectum extract on the suppression of decorin degradation by suppressing Granzyme B. 図５Ｂは、ゲンノショウコエキスとオトギリソウエキスの、グランザイムＢ抑制によるデコリン分解抑制の相乗効果を示した図である。FIG. 5B is a diagram showing the synergistic effect of Geranium thunbergii extract and Hypericum erectum extract on the suppression of decorin degradation by suppressing Granzyme B. 図６Ａは、ゲンノショウコエキスとテンチャエキスの、グランザイムＢ抑制によるデコリン分解抑制の相乗効果を示した図である。FIG. 6A is a diagram showing the synergistic effect of Geranium thunbergii extract and Tencha extract on the suppression of decorin degradation by suppressing Granzyme B. 図６Ｂは、ゲンノショウコエキスとテンチャエキスの、グランザイムＢ抑制によるデコリン分解抑制の相乗効果を示した図である。FIG. 6B is a diagram showing the synergistic effect of Geranium thunbergii extract and Tencha extract on the suppression of decorin degradation by suppressing Granzyme B.

以下、本発明の実施の形態を具体的に説明する。なお、以下で例示する好ましい態様やより好ましい態様等は、「好ましい」や「より好ましい」等の表現にかかわらず適宜相互に組み合わせて使用することができる。また、数値範囲の記載は例示であって、各範囲の上限と下限並びに実施例の数値とを適宜組み合わせた範囲も好ましく使用することができる。さらに、「含有する」又は「含む」等の用語は、「本質的になる」や「のみからなる」と読み替えてもよい。 Hereinafter, embodiments of the present invention will be specifically described. In addition, the preferable embodiment and the more preferable embodiment exemplified below can be used in combination with each other as appropriate regardless of the expressions such as "favorable" and "more preferable". Further, the description of the numerical range is an example, and a range in which the upper limit and the lower limit of each range and the numerical values of the examples are appropriately combined can also be preferably used. Further, terms such as "contains" or "contains" may be read as "essentially" or "consisting only".

〔グランザイムＢ抑制剤〕
以下、本発明のグランザイムＢ抑制剤について詳細に説明する。
グランザイムＢは、細胞傷害性Ｔ細胞及びナチュラルキラー細胞内の細胞質顆粒によって放出されるセリンプロテアーゼであり、様々な細胞外基質を分解し皮膚障害に関与する（Turner et.al., Matrix Biol. (2019) 75-76, 126-140）。なかでも、グランザイムＢがＩ型コラーゲンを保護するデコリンを分解することで、ＭＭＰ−１（matrix metalloproteinase-1）によるＩ型コラーゲン分解が促進されることが知られている（Parkinson et.al., Aging Cell. 2015 Feb; 14(1): 67-77）。グランザイムＢは、通常状態における発現量は低く、皮膚への紫外線照射などに起因して発現誘導される（Hernandez-Pigeon et. al., J Biol Chem. 2006 May 12; 281 (19): 13525-32）。 [Granzyme B inhibitor]
Hereinafter, the Granzyme B inhibitor of the present invention will be described in detail.
Granzyme B is a serine protease released by cytoplasmic granules in cytotoxic T cells and natural killer cells that degrades various extracellular substrates and is involved in skin disorders (Turner et.al., Matrix Biol. (Turner et.al., Matrix Biol.). 2019) 75-76, 126-140). Among them, it is known that Granzyme B decomposes decorin that protects type I collagen, thereby promoting the decomposition of type I collagen by MMP-1 (matrix metalloproteinase-1) (Parkinson et.al., Aging Cell. 2015 Feb; 14 (1): 67-77). Granzyme B has a low expression level under normal conditions and is induced to be expressed by irradiation of the skin with ultraviolet rays (Hernandez-Pigeon et. Al., J Biol Chem. 2006 May 12; 281 (19): 13525- 32).

本発明のグランザイムＢ抑制剤（以下、「抑制剤」と略記する）によるグランザイムＢ抑制の態様としては、グランザイムＢの活性阻害及びグランザイムＢ遺伝子の発現抑制が挙げられる。
グランザイムＢの活性阻害により、コラーゲンが保護される。例えば、理論に縛られないが、グランザイムＢの活性を阻害するとデコリンの分解が抑制され、結果としてＭＭＰ−１によるＩ型コラーゲン分解が抑制される。
グランザイムＢ遺伝子の発現抑制によっても、コラーゲンが保護される。理論に縛られないが、グランザイムＢの発現上昇を抑えることで、より効果的にグランザイムＢの活性を抑制可能である。
しかしながら、本発明の抑制剤によるグランザイムＢの活性阻害及びグランザイムＢ遺伝子の発現抑制、及びそれらによって得られるコラーゲン保護は、必ずしも上記したデコリン分解抑制に関わるものに限定されないことは明らかである。
グランザイムＢは、デコリンの他、様々な細胞外マトリックスを分解することが知られている（Russo et.al., Sci Rep., 2018 Jun 26. 8(1),9690、Turner et.al., Matrix Biol. 2019 Jan, 75-76, 126-140、Pardo et.al., Cell Death Differ., 2007 Oct, 14(10), 1768-79）。例えば、デコリン以外のグランザイムＢの基質として、collagen VII、collagen XVII、α6/β4 integrin、fibronectin、vitronectin、laminin、biglycan、plasminogen、VE-cadherin、ZO-1 及び von Willebrand factor が挙げられる。 Examples of the embodiment of granzyme B inhibition by the granzyme B inhibitor (hereinafter abbreviated as “inhibitor”) of the present invention include inhibition of granzyme B activity and inhibition of Granzyme B gene expression.
Inhibition of Granzyme B activity protects collagen. For example, without being bound by theory, inhibition of the activity of Granzyme B suppresses the degradation of decorin, resulting in the inhibition of type I collagen degradation by MMP-1.
Suppression of the expression of the Granzyme B gene also protects collagen. Although not bound by theory, it is possible to suppress the activity of Granzyme B more effectively by suppressing the increase in the expression of Granzyme B.
However, it is clear that the inhibition of the activity of Granzyme B and the suppression of the expression of the Granzyme B gene by the inhibitor of the present invention, and the collagen protection obtained by them are not necessarily limited to those involved in the above-mentioned inhibition of decorin degradation.
Granzyme B is known to degrade various extracellular matrices in addition to decorin (Russo et.al., Sci Rep., 2018 Jun 26.8 (1), 9690, Turner et.al., Matrix Biol. 2019 Jan, 75-76, 126-140, Pardo et.al., Cell Death Differ., 2007 Oct, 14 (10), 1768-79). Examples of substrates for Granzyme B other than decorin include collagen VII, collagen XVII, α6 / β4 integrin, fibronectin, vitronectin, laminin, biglycan, plasmainogen, VE-cadherin, ZO-1 and von Willebrand factor.

＜ゲンノショウコエキス＞
本発明の抑制剤は、ゲンノショウコエキスを有効成分として含有する。
ゲンノショウコエキスは、ゲンノショウコ（Ｇｅｒａｎｉｕｍ ｔｈｕｎｂｅｒｇｉｉ Ｓｉｅｂｏｌｄ ｅｔ Ｚｕｃｃａｒｉｎｉ（Ｇｅｒａｎｉａｃｅａｅ））から抽出されるエキスである。ゲンノショウコは、フクロソウ科フクロソウ属の多年生植物であり、ミコシグサ、イシャイラズなどとも呼ばれ、整腸作用を有する生薬原料として用いられる。ゲンノショウコエキスは公知物質であり、市場で容易に入手可能であるか、又は調整可能である。ゲンノショウコエキスの調整には当技術分野で周知の方法を使用できるが、例えばゲンノショウコの地上部から、エタノール溶液又は１，３−ブチレングリコール溶液で抽出することで、医薬部外品原料規格２００６に記載のゲンノショウコエキスの規格を満たすものを得ることができる。 <Geranium thunbergii extract>
The inhibitor of the present invention contains Geranium thunbergii extract as an active ingredient.
Geranium thunbergii extract is an extract extracted from Geranium thunbergii Siebold et Zuccarini (Geranium). Geranium thunbergii is a perennial plant belonging to the genus Geranium thunbergii in the family Geranium thunbergii. Geranium thunbergii extract is a known substance and is readily available or adjustable on the market. A method well known in the art can be used for the preparation of Geranium thunbergii extract. For example, by extracting from the above-ground part of Geranium thunbergii with an ethanol solution or a 1,3-butylene glycol solution, it is described in Standards for Raw Materials for Non-pharmaceutical Products 2006. It is possible to obtain a product that meets the specifications of Geranium thunbergii extract.

本発明の抑制剤は、ゲンノショウコエキスを、グランザイムＢ抑制剤の全体積に対して、好ましくは１〜１００μｇ／ｍＬ含有する。本発明の抑制剤は、ゲンノショウコエキスを、グランザイムＢ抑制剤の全体積に対して、好ましくは１．８μｇ／ｍＬ以上含有し、さらに好ましくは５．５μｇ／ｍＬ以上含有し、特に好ましくは１８μｇ／ｍＬ以上含有する。含有量の上限は特に限定されないが、例えば８０μｇ／ｍＬ以下、９０μｇ／ｍＬ以下、９５μｇ／ｍＬ以下で含有する。ゲンノショウコエキスは特異なにおいを有するため、グランザイムＢ抑制剤としての効果を発揮しつつ、ゲンノショウコエキスのにおいを抑えることが可能な含有量としては、例えば１８〜５５μｇ／ｍＬが望ましい。 The inhibitor of the present invention contains geranium thunbergii extract, preferably 1 to 100 μg / mL, based on the total volume of the granzyme B inhibitor. The inhibitor of the present invention contains geranium thunbergii extract, preferably 1.8 μg / mL or more, more preferably 5.5 μg / mL or more, and particularly preferably 18 μg / mL, based on the total volume of the granzyme B inhibitor. Contains more than mL. The upper limit of the content is not particularly limited, but the content is, for example, 80 μg / mL or less, 90 μg / mL or less, and 95 μg / mL or less. Since Geranium thunbergii extract has a peculiar odor, for example, 18 to 55 μg / mL is desirable as the content capable of suppressing the odor of Geranium thunbergii extract while exhibiting the effect as a granzyme B inhibitor.

＜追加の植物エキス＞
本発明の抑制剤は、ゲンノショウコエキスに加え、有効成分としてグランザイムＢ抑制作用を有する１種以上の植物エキス（以下、「追加の植物エキス」と記載する）をさらに含有していてもよい。
グランザイムＢ抑制作用を有する追加の植物エキスとしては、限定されるものではないが、例えば実施例の〔蛍光基質を用いたグランザイムＢ活性阻害効果の評価系〕に従い、グランザイムＢ阻害活性（IC50値<３μｇ／ｍＬ）を示すものを選択することができる。 <Additional plant extract>
In addition to the geranium thunbergii extract, the inhibitor of the present invention may further contain one or more plant extracts having a granzyme B inhibitory action (hereinafter referred to as "additional plant extract") as an active ingredient.
The additional plant extract having a granzyme B inhibitory effect is not limited, but is, for example, according to the [Evaluation system of the granzyme B activity inhibitory effect using a fluorescent substrate] of the example, the granzyme B inhibitory activity (IC 50 value). Those showing <3 μg / mL) can be selected.

追加の植物エキスの例としては、ザクロ果皮エキス、イエローヒマラヤンラズベリー根エキス、ユーカリエキス、ワレモコウエキス、イロハモミジ葉エキス、シモツケソウエキス、ハマメリスエキス、バラエキス、アボガドエキス、テンチャエキス、メマツヨイグサ抽出液、ウーロン茶エキス、カシア樹皮エキス、ユキノシタエキス、メリッサエキス、トルメンチラエキス、ダマスクバラ花エキス、ヨモギエキス、ヒキオコシエキス、コンフリーエキス、オトギリソウエキス、セイヨウハッカエキス、ブドウ葉エキス、シナノキエキス、エイジツエキス、ビワ葉エキス、海藻エキス、シャクヤクエキス、ドクダミエキス、サンザシエキス、シソエキス、ラベンダーエキス、ノバラエキス、アセンヤクエキス、サガラメエキス、ボタンエキス、アスパラサスリネアリスエキス、カンゾウ抽出末及びクララエキスからなる群から選択されるものが挙げられる。これらのエキスは、単独又は組み合わせのいずれであっても、追加の植物エキスとして用いることができる。 Examples of additional plant extracts include pomegranate peel extract, yellow Himalayan raspberry root extract, eucalyptus extract, crackle extract, Irohamomiji leaf extract, Shimotsukesou extract, Hamamelis extract, rose extract, avocado extract, tencha extract, mematsuyoigusa extract, oolong tea extract, Cassia bark extract, Yukinoshita extract, Melissa extract, Tormentilla extract, Damask rose flower extract, Yomogi extract, Hikiokoshi extract, Confree extract, Otogirisou extract, Seiyouhakka extract, Grape leaf extract, Shinanoki extract, Agetsu extract, Biwa leaf extract , Seaweed extract, Shakuyaku extract, Dokudami extract, Sanzashi extract, Perilla extract, Lavender extract, Novara extract, Asenyaku extract, Sagarame extract, Button extract, Asparasas linearis extract, Kanzo extract powder and Clara extract. Can be mentioned. These extracts, either alone or in combination, can be used as additional plant extracts.

追加の植物エキスは、単独でもグランザイムＢの抑制作用を有するが、ゲンノショウコエキスと組み合わせると相乗的なグランザイムＢ抑制作用を発揮するものが好ましい。ゲンノショウコエキスとの相乗効果を発揮する追加の植物エキスとしては、オトギリソウセキス及び／又はテンチャエキスが挙げられる。 Although the additional plant extract alone has an inhibitory effect on granzyme B, it is preferably one that exhibits a synergistic inhibitory effect on granzyme B when combined with the geranium thunbergii extract. Additional plant extracts that exert a synergistic effect with Geranium thunbergii extract include Hypericum erectum and / or Tencha extract.

＜オトギリソウエキス＞
オトギリソウエキスは、セイヨウオトギリソウ（Ｈｙｐｅｒｉｃｕｍ ｐｅｒｆｏｒａｔｕｍ Ｌｉｎｎｅ）又はオトギリソウ（Ｈｙｐｅｒｉｃｕｍ ｅｒｅｃｔｕｍ Ｔｈｕｎｂｅｒｇ（Ｇｕｔｔｉｆｅｒａｅ））から抽出されるエキスである。セイヨウオトギリソウ及びオトギリソウは、オトギリソウ科オトギリソウ属の多年生植物である。オトギリソウエキスは公知物質であり、市場で容易に入手可能であるか、又は調製可能である。オトギリソウエキスの調整には当技術分野で周知の方法を使用できるが、例えばセイヨウオトギリソウ又はオトギリソウの地上部から、水、エタノール、プロピレングリコール、１，３−ブチレングリコール、ジエチレングリコール、又はこれらの混液により抽出することで、医薬部外品原料規格２００６に記載のオトギリソウエキスの規格を満たすものを得ることができる。 <Hypericum extract>
Hypericum extract is an extract extracted from Hypericum perforatum Line or Hypericum erectum Thumberg (Guttipherae). Hypericum and St. John's wort are perennial plants of the genus Hypericum in the family Hypericum. Hypericum extract is a known substance and is readily available on the market or can be prepared. Methods well known in the art can be used to prepare the Hypericum extract, for example, extracted from above-ground parts of Hypericum erectum or Hypericum erectum with water, ethanol, propylene glycol, 1,3-butylene glycol, diethylene glycol, or a mixture thereof. By doing so, it is possible to obtain a product that satisfies the specifications of Hypericum erectum extract described in the standard for raw materials for non-pharmaceutical products 2006.

＜テンチャエキス＞
テンチャエキスは、テンヨウケンコウシ（Ｒｕｂｕｓ ｓｕａｖｉｓｓｉｍｕｓ Ｓｈｕｇａｎ Ｌｅｅ．（Ｒｏｓａｃｅａｅ））から抽出されるエキスである。テンヨウケンコウシは、バラ科キイチゴ属の耐寒性落葉低木である。テンチャエキスは公知物質であり、市場で容易に入手可能であるか、又は調製可能である。テンチャエキスの調整には当技術分野で周知の方法を使用できるが、例えばテンヨウケンコウシの葉を熱湯に浸漬した後、葉を乾燥し、さらに加熱したものを熱湯にて抽出することで医薬部外品原料規格２００６に記載のテンチャエキスの規格を満たすものを得ることができる。また、抽出溶媒としては１，３−ブチレングリコールを用いることもできる。 <Tencha extract>
The tencha extract is an extract extracted from Rubus suavissimus Shugan Lee. (Rosaceae). Tenyoukenkoushi is a cold-tolerant deciduous shrub of the genus Rubus in the family Rosaceae. Tencha extract is a known substance and is readily available or available on the market. A method well known in the art can be used to prepare the tencha extract. For example, the leaves of Tenyokenkoushi are soaked in boiling water, the leaves are dried, and the heated one is extracted with boiling water to extract the quasi-drug. It is possible to obtain a product that meets the specifications of the tencha extract described in the quasi-drug raw material standard 2006. Further, 1,3-butylene glycol can also be used as the extraction solvent.

オトギリソウエキス及びテンチャエキスのほか、追加の植物エキスの各々は公知物質であり、市場で容易に入手可能であるか、又は当技術分野で周知の方法を使用して調製可能である。 In addition to Hypericum erectum extract and Tencha extract, each of the additional plant extracts is a known substance and is readily available on the market or can be prepared using methods well known in the art.

本発明の抑制剤に配合される追加の植物エキスの量は特に限定されないが、例えば追加の植物エキスとゲンノショウコエキスの質量比を１：１〜１：４で配合した場合に、特に高い効率でグランザイムＢを抑制することができる。追加の植物エキスとゲンノショウコエキスの質量比は、例えば１：１．５であり、好ましくは１：２であり、さらに好ましくは１：３であり、特に好ましくは１：４である。
オトギリソウエキスが追加の植物エキスとして配合される場合、オトギリソウエキスとゲンノショウコエキスの質量比は、好ましくは１：１〜１：４であり、当該濃度比で高い相乗効果が得られる。
テンチャエキスが追加の植物エキスとして配合される場合、テンチャエキスとゲンノショウコエキスの質量比は、好ましくは１：１〜１：４であり、当該濃度比で高い相乗効果が得られる。 The amount of the additional plant extract blended in the inhibitor of the present invention is not particularly limited, but for example, when the mass ratio of the additional plant extract and the geranium thunbergii extract is blended at a ratio of 1: 1 to 1: 4, the efficiency is particularly high. Granzyme B can be suppressed. The mass ratio of the additional plant extract to the geranium thunbergii extract is, for example, 1: 1.5, preferably 1: 2, more preferably 1: 3, and particularly preferably 1: 4.
When the Hypericum erectum extract is blended as an additional plant extract, the mass ratio of the Hypericum erectum extract and the Geranium thunbergii extract is preferably 1: 1 to 1: 4, and a high synergistic effect can be obtained at the concentration ratio.
When the tencha extract is blended as an additional plant extract, the mass ratio of the tencha extract and the geranium thunbergii extract is preferably 1: 1 to 1: 4, and a high synergistic effect can be obtained at the concentration ratio.

本発明の抑制剤は、追加の植物エキスを、抑制剤の全体積に対して例えば１〜１００μｇ／ｍＬ含有する。
追加の植物エキスとしてオトギリソウエキスを含有する場合、オトギリソウエキスを、抑制剤の全体積に対して、好ましくは１．５μｇ／ｍＬ以上含有し、さらに好ましくは４．６μｇ／ｍＬ以上含有し、特に好ましくは１５μｇ／ｍＬ以上含有する。含有量の上限は特に限定されないが、例えば８０μｇ／ｍＬ以下、９０μｇ／ｍＬ以下、９５μｇ／ｍＬ以下で含有する。当該濃度範囲で配合されると、抑制剤のより高いグランザイムＢ活性阻害作用が発揮される。
追加の植物エキスとしてテンチャエキスを含有する場合、テンチャエキスを、抑制剤の全体積に対して、好ましくは３．１μｇ／ｍＬ以上含有し、さらに好ましくは１０μｇ／ｍＬ以上含有し、特に好ましくは３１μｇ／ｍＬ以上含有する。含有量の上限は特に限定されないが、例えば８０μｇ／ｍＬ以下、９０μｇ／ｍＬ以下、９５μｇ／ｍＬ以下で含有する。当該濃度範囲で配合されると、抑制剤のより高いグランザイムＢ活性阻害作用が発揮される。 The inhibitor of the present invention contains an additional plant extract, eg, 1-100 μg / mL, relative to the total volume of the inhibitor.
When Hypericum erectum extract is contained as an additional plant extract, the Hypericum erectum extract is preferably contained in an amount of 1.5 μg / mL or more, more preferably 4.6 μg / mL or more, particularly preferably with respect to the total volume of the inhibitor. Contains 15 μg / mL or more. The upper limit of the content is not particularly limited, but the content is, for example, 80 μg / mL or less, 90 μg / mL or less, and 95 μg / mL or less. When blended in the concentration range, a higher granzyme B activity inhibitory effect of the inhibitor is exhibited.
When the tencha extract is contained as an additional plant extract, the tencha extract is preferably contained in an amount of 3.1 μg / mL or more, more preferably 10 μg / mL or more, and particularly preferably 31 μg, based on the total volume of the inhibitor. Contains more than / mL. The upper limit of the content is not particularly limited, but the content is, for example, 80 μg / mL or less, 90 μg / mL or less, and 95 μg / mL or less. When blended in the concentration range, a higher granzyme B activity inhibitory effect of the inhibitor is exhibited.

＜任意成分＞
本発明の抑制剤には、ゲンノショウコエキス及び追加の植物エキスに加え、本発明の効果を損なわない範囲で、１種以上の成分を任意に配合できる。また、任意成分は、後述する化粧料組成物に適用可能な成分であってもよい。任意成分としては、以下に示すものが挙げられる。

〔基剤成分〕
水（例えば、精製水）、低級アルコール（例えば、エタノール）、多価アルコール（例えば、プロピレングリコール）、油脂、界面活性剤、紫外線吸収剤、紫外線散乱剤、増粘剤、色素等

〔その他〕
乳化剤、香料、防腐剤等

〔保湿成分〕
トコフェロール、ヒアルロン酸Ｎａ、ミツバアケビ茎エキス、加水分解エラスチン、加水分解コラーゲン、アカヤジオウ根エキス、アセチルヒアルロン酸Ｎａ、アルニカ花エキス、アロエベラ葉エキス、オウゴンエキス、オーキッドエキス、オタネニンジン根エキス、カミツレ花エキス、キュウリ果実エキス、クズ根エキス、クロレラエキス、セイヨウキズタ葉／茎エキス、セイヨウシロヤナギ樹皮エキス、セイヨウニワトコ花エキス、ゼニアオイエキス、センブリエキス、ダイズエキス、トウキンセンカ花エキス、トリスヘキシルデカン酸ピリドキシン、パリエタリアエキス、パルミチン酸レチノール、フユボダイジュ花エキス、マクロプチリウムアトロプルプレウム花／葉／茎エキス、ヤグルマギク花エキス、ローマカミツレ花エキス、ローヤルゼリーエキス、加水分解ハトムギ種子、オウレン根茎エキス、カワラヨモギ花エキス、キハダ樹皮エキス、クチナシ果実エキス、グリチルリチン酸２Ｋ、ゲンチアナ根茎／根エキス、センキュウ根茎エキス、トウキ根エキス、ドクダミエキス、ナツメ果実エキス、ハトムギ種子エキス、ベタイン、ホホバエステル、加水分解ヒアルロン酸、水添レチノール、水溶性プロテオグリカン <Arbitrary ingredient>
In addition to the geranium thunbergii extract and the additional plant extract, the inhibitor of the present invention may optionally contain one or more components as long as the effects of the present invention are not impaired. Further, the optional ingredient may be an ingredient applicable to the cosmetic composition described later. Examples of the optional component include those shown below.

[Base ingredient]
Water (eg purified water), lower alcohols (eg ethanol), polyhydric alcohols (eg propylene glycol), fats and oils, surfactants, UV absorbers, UV scatterers, thickeners, pigments, etc.

〔others〕
Emulsifiers, fragrances, preservatives, etc.

[Moisturizing ingredients]
Tocopherol, Na hyaluronate, honeybee stalk extract, hydrolyzed elastin, hydrolyzed collagen, red sardine root extract, Na acetyl hyaluronate, Arnica flower extract, aloe vera leaf extract, ginger extract, orchid extract, octopus carrot root extract, chamomile flower extract, cucumber Fruit extract, kudzu root extract, chlorella extract, sardine leaf / stem extract, sardine bark extract, sardine flower extract, zenia oi extract, senburi extract, soybean extract, tokinsenka flower extract, trishexyldecanoic acid pyridoxin, parishetaria extract, palmitin Retinol acid, Fuyubodaiju flower extract, Macroputilium atropulpreum flower / leaf / stem extract, Yagurumagiku flower extract, Roman chamomile flower extract, Royal jelly extract, Hydrolyzed pigeon seeds, Ouren root stem extract, Kawarayomogi flower extract, Kihada bark extract , Kuchinashi fruit extract, glycyrrhizinic acid 2K, Gentian rhizome / root extract, Senkyu rhizome extract, Touki root extract, Dokudami extract, Natsume fruit extract, Hatomugi seed extract, betaine, jojoba ester, hydrolyzed hyaluronic acid, hydrogenated retinol, water-soluble Proteoglycan

＜剤型＞
本発明の抑制剤の剤型は、特に限定されない。例えば、溶液系、可溶化系、乳化系、粉末分散系、水−油二層系、水−油−粉末三層系、軟膏、ゲル、及びエアゾール等が挙げられる。本発明の抑制剤は、上記した任意成分を適宜組み合わせて配合することにより、所望の剤型とすることができる。 <Dosage form>
The dosage form of the inhibitor of the present invention is not particularly limited. For example, solution system, solubilization system, emulsification system, powder dispersion system, water-oil two-layer system, water-oil-powder three-layer system, ointment, gel, aerosol and the like can be mentioned. The inhibitor of the present invention can be made into a desired dosage form by appropriately combining and blending the above-mentioned arbitrary components.

＜製造方法＞
本発明の抑制剤は、当技術分野で通常用いられる製法に従い製造することができる。製法としては、例えばスキンケア製品（皮膚用化粧料）の一般的な製法を挙げることができるが、これに限定されない。例えば、有効成分を基材成分へ添加し、混合することで製造できる。 <Manufacturing method>
The inhibitor of the present invention can be produced according to a production method usually used in the art. Examples of the manufacturing method include, but are not limited to, a general manufacturing method for skin care products (skin cosmetics). For example, it can be produced by adding an active ingredient to a base material component and mixing them.

＜使用方法＞
本発明の抑制剤は、塗布などにより対象に適用することができる。本発明の抑制剤の適用対象は、概して哺乳類であり、特にヒトが想定される。 <How to use>
The inhibitor of the present invention can be applied to a subject by coating or the like. The target of application of the inhibitor of the present invention is generally mammals, and humans are particularly assumed.

〔コラーゲン分解抑制剤〕
ゲンノショウコエキスは、上述したようにグランザイムＢ活性阻害作用及びグランザイムＢ遺伝子発現抑制作用により、コラーゲン分解を抑制する。したがって、本発明のグランザイムＢ抑制剤は、コラーゲン分解抑制剤としても把握できる。
本発明のコラーゲン分解抑制剤によるコラーゲン分解抑制の態様としては、グランザイムＢの活性阻害及びグランザイムＢ遺伝子の発現抑制が挙げられる。
本発明のコラーゲン分解抑制剤の有効成分、任意成分、剤型、製造方法、及び使用方法は、グランザイムＢ抑制剤について述べた説明が適用される。 [Collagen decomposition inhibitor]
As described above, Geranium thunbergii suppresses collagen degradation by inhibiting granzyme B activity and suppressing granzyme B gene expression. Therefore, the Granzyme B inhibitor of the present invention can also be grasped as a collagen degradation inhibitor.
Examples of the mode of suppressing collagen decomposition by the collagen decomposition inhibitor of the present invention include inhibition of the activity of Granzyme B and suppression of the expression of the Granzyme B gene.
The description of the Granzyme B inhibitor is applied to the active ingredient, optional ingredient, dosage form, production method, and usage method of the collagen decomposition inhibitor of the present invention.

〔化粧料組成物〕
本発明の化粧料組成物は、グランザイムＢ抑制剤のみからなる。本発明の化粧料組成物の使用形態は、特に限定されない。例えば、化粧水、クリーム、乳液、エッセンス、ゼリー、ジェル、軟膏、ファンデーション、及びパック等が挙げられる。
本発明の化粧料組成物は、コラーゲンの維持を必要とする対象の皮膚に、塗布などにより適用することができる。本発明のグランザイムＢ抑制剤の適用対象は、概して哺乳類であり、特にヒトが想定される。
本発明の化粧料組成物の有効成分、任意成分、剤型、及び製造方法は、グランザイムＢ抑制剤について述べた説明が適用される。 [Cosmetic composition]
The cosmetic composition of the present invention comprises only a granzyme B inhibitor. The form of use of the cosmetic composition of the present invention is not particularly limited. Examples include lotions, creams, emulsions, essences, jellies, gels, ointments, foundations, facial masks and the like.
The cosmetic composition of the present invention can be applied to the skin of a subject in need of collagen maintenance by application or the like. The target of application of the Granzyme B inhibitor of the present invention is generally mammals, and humans in particular are assumed.
The description of the Granzyme B inhibitor is applied to the active ingredient, optional ingredient, dosage form, and manufacturing method of the cosmetic composition of the present invention.

〔医薬組成物〕
本発明のグランザイムＢ抑制剤は、医薬用途にも利用可能である。対象となる疾患には創傷、火傷、自己免疫性皮膚炎、慢性創傷、ケロイド、肥厚性瘢痕、円板状エリテマトーデス、円形脱毛症、強皮症、スティーブンス・ジョンソン症候群、中毒性表皮壊死症などが挙げられ、本発明のグランザイムＢ抑制剤からなる医薬組成物を、塗布などにより対象に投与することで当該疾患の症状を改善することができる。医薬組成物の適用対象は概して哺乳類であり、特にヒトが想定される。
医薬組成物の有効成分、任意成分、剤型、及び製造方法は、グランザイムＢ抑制剤について述べた説明が適用される。 [Pharmaceutical composition]
The Granzyme B inhibitor of the present invention can also be used for pharmaceutical purposes. Target diseases include wounds, burns, autoimmune dermatitis, chronic wounds, keloids, hypertrophic scars, discoid lupus erythematosus, discoid lupus erythematosus, sclerosis, Stevens Johnson syndrome, toxic epidermal necrolysis, etc. The symptoms of the disease can be improved by administering the pharmaceutical composition comprising the Granzyme B inhibitor of the present invention to the subject by application or the like. The target of application of the pharmaceutical composition is generally mammals, and humans are particularly envisioned.
The description of the Granzyme B inhibitor applies to the active ingredient, optional ingredient, dosage form, and manufacturing method of the pharmaceutical composition.

次に、実施例により本発明の効果を具体的に説明するが、本発明は実施例に限定されるものではない。 Next, the effect of the present invention will be specifically described with reference to Examples, but the present invention is not limited to the Examples.

〔植物エキスの調整〕
下記のように、各植物エキスを調整した。
ゲンノショウコエキスを、医薬部外品原料規格２００６に記載される方法により調製し、以下の実験に用いた。なお、抽出にはゲンノショウコの地上部（花、葉、及び茎）を用い、抽出溶媒としては１，３−ブチレングリコール溶液を使用した。
オトギリソウエキスを、医薬部外品原料規格２００６に記載される方法により調製し、以下の実験に用いた。なお、抽出にはセイヨウオトギリソウの地上部（花、葉、及び茎）を用い、抽出溶媒としては１，３−ブチレングリコール溶液を使用した。
テンチャエキスを、医薬部外品原料規格２００６に記載される方法により調製し、以下の実験に用いた。なお、抽出にはテンチャの葉を用い、抽出溶媒としては熱湯を使用した。 [Adjustment of plant extract]
Each plant extract was adjusted as follows.
Geranium thunbergii extract was prepared by the method described in Quasi-drug Raw Material Standard 2006 and used in the following experiments. The above-ground part of Geranium thunbergii (flowers, leaves, and stems) was used for extraction, and a 1,3-butylene glycol solution was used as the extraction solvent.
Hypericum extract was prepared by the method described in Quasi-drug Raw Material Standard 2006 and used in the following experiments. The above-ground part (flowers, leaves, and stems) of St. John's wort was used for extraction, and a 1,3-butylene glycol solution was used as the extraction solvent.
Tencha extract was prepared by the method described in Quasi-drug Raw Material Standard 2006 and used in the following experiments. Tencha leaves were used for extraction, and boiling water was used as the extraction solvent.

〔ヒト表皮角化細胞を使用したアッセイ系〕
下記の手順に従い、ヒト表皮角化細胞を培養し、ヒト表皮単層モデルを作成した。
ヒト表皮角化細胞を、37℃、5% CO2 インキュベーター内、75 cm2フラスコ中で、10% FBS, 1% PS mix含有D-MEM (High Glucose) with L-Glutamine and Phenol Red（富士フイルム和光純薬社製）、以下D-MEM培地を用いて70〜90%コンフルエントまで培養した。Accutase Solution（Sigma-Aldrich社製）を用いて細胞を剥離し、24 well clear plate（Corning社製）に1.5 x 104 cells/cm2となるように細胞を播種した。D-MEM培地を用いて37℃、5% CO2インキュベーター内で３日間培養した細胞をヒト表皮単層モデルとし、アッセイに使用した。 [A assay system using human epidermal keratinocytes]
Human epidermal keratinized cells were cultured according to the following procedure to create a human epidermal monolayer model.
Human epidermal keratinized cells in a 75 cm 2 flask at 37 ° C in a 5% CO 2 incubator, containing 10% FBS, 1% PS mix D-MEM (High Glucose) with L-Glutamine and Phenol Red (Fujifilm) Wako Pure Chemical Industries, Ltd.), cultivated to 70-90% confluent using D-MEM medium. The cells were detached using an Accutase Solution (manufactured by Sigma-Aldrich), and the cells were seeded on a 24-well clear plate (manufactured by Corning) to 1.5 x 104 cells / cm 2. Cells cultured in D-MEM medium at 37 ° C. in a 5% CO 2 incubator for 3 days were used as a human epidermal monolayer model for the assay.

〔グランザイムＢ遺伝子の発現抑制効果の評価系〕
通常状態の皮膚ではグランザイムＢ遺伝子の発現量は低く、紫外線（UV-B）照射によって発現誘導されることが知られている。そこで、UV-B 照射したヒト表皮単層モデルにゲンノショウコエキスを添加し、UV-B 照射によるグランザイムＢ遺伝子発現誘導の抑制効果を評価した。
ヒト表皮単層モデルにUV-B（波長：302 nm, 積算照射量：10 mJ/cm2）照射後、D-MEM中に溶解させたゲンノショウコエキスを添加し、16時間培養後の試験細胞から遺伝子を抽出し、Real-time PCR法にてmRNA量を測定した。グランザイムＢ遺伝子発現量は、GAPDHを内標準物質としたΔΔCt法を用いて算出した。
ゲンノショウコエキスによるグランザイムＢ遺伝子の発現抑制効果は、UV-B非照射群を対照とした% of control値を算出し、UV-B 照射したゲンノショウコエキス非処置群と処置群を有意差検定（Dunnett's Multiple Comparison Test）にて比較することで評価した。 [Evaluation system for suppressing the expression of Granzyme B gene]
It is known that the expression level of the granzyme B gene is low in the skin under normal conditions, and the expression is induced by irradiation with ultraviolet rays (UV-B). Therefore, Geranium thunbergii extract was added to the UV-B-irradiated human epidermis monolayer model, and the inhibitory effect of UV-B irradiation on the induction of Granzyme B gene expression was evaluated.
After UV-B (wavelength: 302 nm, cumulative irradiation dose: 10 mJ / cm 2 ) irradiation to the human epidermis monolayer model, gennoshoco extract dissolved in D-MEM was added, and from the test cells after culturing for 16 hours. The gene was extracted and the amount of mRNA was measured by the real-time PCR method. The expression level of the Granzyme B gene was calculated using the ΔΔCt method using GAPDH as an internal standard substance.
For the effect of suppressing the expression of the Granzyme B gene by Geranium thunbergii extract, a% of control value was calculated using the UV-B non-irradiated group as a control, and a significant difference test (Dunnett's Multiple) was performed between the UV-B-irradiated Geranium thunbergii extract untreated group and the treated group. It was evaluated by comparing with Comparison Test).

〔ゲンノショウコエキスによるグランザイムＢ遺伝子発現抑制効果〕
ゲンノショウコエキスによるグランザイムＢ遺伝子発現抑制効果を、図１に示す。
図１に示されるように、ゲンノショウコエキスがUV-B 刺激誘導性のグランザイムＢ遺伝子発現上昇を抑制した。 [Granzyme B gene expression inhibitory effect of Geranium thunbergii extract]
The effect of suppressing the expression of the Granzyme B gene by Geranium thunbergii extract is shown in FIG.
As shown in FIG. 1, Geranium thunbergii extract suppressed UV-B stimulation-induced upregulation of Granzyme B gene expression.

〔デコリンを用いたグランザイムＢ活性阻害効果の評価系〕
グランザイムＢは、様々な細胞外基質を分解し皮膚障害に関与する。なかでも、グランザイムＢがデコリンを分解することで、MMP-1によるＩ型コラーゲン分解が促進されることが知られている。そこで、デコリンに植物エキス及びグランザイムＢを添加し、植物エキスによるグランザイムＢ活性阻害効果を評価した。
グランザイムＢリコンビナント（終濃度 100 ng/μL、R&D systems社製）、カテプシンＣリコンビナント（終濃度 10 ng/μL、R&D systems社製）、CHAPS（終濃度0.01%）、DTT（終濃度5 mM）、NaCl（終濃度50 mM）、MES（終濃度50 mM, pH 5.5）を混合し、37℃で16時間反応させ、グランザイムＢを活性化した。デコリンリコンビナント（終濃度 10 ng/μL、Abcam社製）、植物エキス、活性化したグランザイムＢ（終濃度 2.5 ng/μL）及びTris-HCl（終濃度 50 mM, pH 7.5)を混合し、37℃、20〜22時間反応させた。反応後の溶液をSDS-PAGEにてタンパク分離した後、銀染色試薬（コスモバイオ社製）でタンパク質を検出し、デコリン分解物断片のタンパク量を定量化した。
グランザイムＢ活性阻害効果は、植物エキス非処置群を対照とした% of control値を算出し、植物エキス非処置群と処置群を有意差検定（Dunnett's Multiple Comparison Test）にて比較することで評価した。 [Evaluation system for the inhibitory effect of Granzyme B activity using decorin]
Granzyme B degrades various extracellular matrices and is involved in skin disorders. Among them, it is known that Granzyme B decomposes decorin to promote the decomposition of type I collagen by MMP-1. Therefore, a plant extract and Granzyme B were added to decorin, and the effect of the plant extract on inhibiting Granzyme B activity was evaluated.
Granzyme B recombinant (final concentration 100 ng / μL, manufactured by R & D systems), cathepsin C recombinant (final concentration 10 ng / μL, manufactured by R & D systems), CHAPS (final concentration 0.01%), DTT (final concentration 5 mM), NaCl (final concentration 50 mM) and MES (final concentration 50 mM, pH 5.5) were mixed and reacted at 37 ° C. for 16 hours to activate Granzyme B. A mixture of decorin recombinant (final concentration 10 ng / μL, manufactured by Abcam), plant extract, activated granzyme B (final concentration 2.5 ng / μL) and Tris-HCl (final concentration 50 mM, pH 7.5) at 37 ° C. , 20-22 hours reaction. After protein separation of the reaction solution by SDS-PAGE, protein was detected by silver staining reagent (manufactured by Cosmo Bio Co., Ltd.), and the amount of protein in the decorinated fragment was quantified.
The effect of inhibiting Granzyme B activity was evaluated by calculating the% of control value using the plant extract untreated group as a control and comparing the plant extract untreated group and the treated group by the Dunnett's Multiple Comparison Test. ..

〔ゲンノショウコエキスによるグランザイムＢ活性阻害効果〕
上記デコリンを用いたグランザイムＢ活性阻害効果の評価系において、植物エキスとしてゲンノショウコエキスを処置し、グランザイムＢ活性阻害を評価した。評価結果を図２に示す。
図２に示されるように、ゲンノショウコエキスがグランザイムＢの活性を阻害することで、デコリンの分解を抑制した。 [Granzyme B activity inhibitory effect of Geranium thunbergii extract]
In the evaluation system of the effect of inhibiting Granzyme B activity using the above-mentioned decorin, Geranium thunbergii extract was treated as a plant extract to evaluate the inhibition of Granzyme B activity. The evaluation results are shown in FIG.
As shown in FIG. 2, Geranium thunbergii extract inhibited the decomposition of decorin by inhibiting the activity of Granzyme B.

〔蛍光基質を用いたグランザイムＢ活性阻害効果の評価系〕
蛍光基質としてN-Acetyl-Ile-Glu-Pro-Asp-7-amido-4-methylcoumarin（Ac-IEPD-AMC, Sigma-Aldrich）を使用して、図３に示す植物エキスのほか、数多くの植物エキスに関してグランザイムＢ活性阻害効果を評価した。
グランザイムＢリコンビナントは、デコリンを用いたグランザイムＢ活性阻害効果の評価系と同じ手順で活性化した。Ac-IEPD-AMC（終濃度50μM）、植物エキス、活性化したグランザイムB（終濃度 0.3 ng/μL）、CHAPS（終濃度0.01%）及びHEPES（終濃度50 mM, pH 7.5)を、384 well black plate（Corning社製）に添加し、30℃、60分間反応させた。溶液添加直後（R1）と反応後（R2）に蛍光強度（ex/em = 380/460 nm）を測定した。
グランザイムＢ活性は、下記計算式を用いて算出した。
% of control (%) = 植物エキス処置群の蛍光値 (R2-R1) / 植物エキス非処置群の蛍光値 (R2-R1) x 100
グランザイムB活性阻害効果は、% of control 値から50% 阻害濃度（IC50値）を算出することで評価した。 [Evaluation system for the effect of inhibiting Granzyme B activity using a fluorescent substrate]
Using N-Acetyl-Ile-Glu-Pro-Asp-7-amido-4-methylcoumarin (Ac-IEPD-AMC, Sigma-Aldrich) as the fluorescent substrate, in addition to the plant extracts shown in FIG. 3, many plants The effect of inhibiting Granzyme B activity was evaluated for the extract.
Granzyme B recombinant was activated by the same procedure as the evaluation system for the effect of inhibiting Granzyme B activity using decorin. Ac-IEPD-AMC (final concentration 50 μM), plant extract, activated granzyme B (final concentration 0.3 ng / μL), CHAPS (final concentration 0.01%) and HEPES (final concentration 50 mM, pH 7.5), 384 well It was added to a black plate (manufactured by Corning) and reacted at 30 ° C. for 60 minutes. Fluorescence intensity (ex / em = 380/460 nm) was measured immediately after the solution was added (R1) and after the reaction (R2).
Granzyme B activity was calculated using the following formula.
% of control (%) = Fluorescence value of plant extract treated group (R2-R1) / Fluorescence value of plant extract untreated group (R2-R1) x 100
The effect of inhibiting Granzyme B activity was evaluated by calculating the 50% inhibition concentration (IC 50 value) from the% of control value.

〔各種植物エキスによるグランザイムＢ活性阻害効果〕
上記蛍光基質を用いたグランザイムＢ活性阻害効果の評価系において、グランザイムＢ阻害活性（IC50値<３μｇ／ｍＬ）を示した植物エキスを、図３に示す。
図３に示すように、ゲンノショウコエキス以外の複数の植物エキスが、グランザイムＢの活性を阻害した。 [Granzyme B activity inhibitory effect of various plant extracts]
FIG. 3 shows a plant extract showing granzyme B inhibitory activity (IC 50 value <3 μg / mL) in the evaluation system of the granzyme B activity inhibitory effect using the fluorescent substrate.
As shown in FIG. 3, a plurality of plant extracts other than Geranium thunbergii extract inhibited the activity of Granzyme B.

〔オトギリソウエキス及びテンチャエキスによるグランザイムＢ活性阻害効果〕
上記デコリンを用いたグランザイムＢの活性阻害効果の評価系において、オトギリソウエキス及びテンチャエキスを植物エキスとしてそれぞれ処置し、グランザイムＢ活性阻害を評価した。評価結果を図４に示す。
図４に示されるように、オトギリソウエキス及びテンチャエキスもまた、ゲンノショウコエキスと同様に、グランザイムＢの活性を阻害することでデコリンの分解を抑制した。 [Granzyme B activity inhibitory effect of Hypericum extract and Tencha extract]
In the evaluation system of the activity inhibitory effect of Granzyme B using the above decorin, Hypericum erectum extract and Tencha extract were treated as plant extracts, respectively, and the inhibition of Granzyme B activity was evaluated. The evaluation results are shown in FIG.
As shown in FIG. 4, Hypericum erectum extract and Tencha extract, like Geranium thunbergii extract, also suppressed the degradation of decorin by inhibiting the activity of Granzyme B.

〔グランザイムＢ阻害によるデコリン分解抑制の相乗効果の評価系〕
試験は、デコリンを用いたグランザイムＢの活性阻害効果の評価系と同じ手順で実施した。
グランザイムＢ阻害によるデコリン分解抑制効果は、植物エキス非処置群を対照とした% of control値を算出し、各群を有意差検定（Tukey's Multiple Comparison Test）にて比較することで評価した。
相乗効果は、デコリン分解抑制率（100 - % of control 値 (%)）を算出し、植物エキス同時処置したデコリン分解抑制率の実測値から、植物エキス単独処置した各々のデコリン分解抑制率を足し合わせた理論値の比率を算出し評価した。
例：
相乗効果 = 植物エキスAとBを同時処置した際のデコリン分解抑制率（実測値）
／ ［ 植物エキスAを単独処置した際のデコリン分解抑制率 + 植物エキスBを単独処置した際のデコリン分解抑制率］（理論値） [Evaluation system for the synergistic effect of suppressing decorin degradation by inhibiting Granzyme B]
The test was carried out in the same procedure as the evaluation system for the activity inhibitory effect of Granzyme B using decorin.
The effect of inhibiting granzyme B on decorin degradation was evaluated by calculating the% of control value using the plant extract non-treated group as a control and comparing each group with a significant difference test (Tukey's Multiple Comparison Test).
For the synergistic effect, the decorin decomposition suppression rate (100-% of control value (%)) is calculated, and the measured value of the decorin decomposition suppression rate treated with the plant extract is added to the respective decorin decomposition suppression rates treated with the plant extract alone. The ratio of the combined theoretical values was calculated and evaluated.
example:
Synergistic effect = Suppression rate of decorin degradation when plant extracts A and B are treated at the same time (measured value)
/ [Decorin degradation inhibitory rate when plant extract A is treated alone + decorin degradation inhibitory rate when plant extract B is treated alone] (theoretical value)

〔ゲンノショウコエキスとオトギリソウエキスのデコリン分解抑制における相乗効果〕
ゲンノショウコエキスとオトギリソウエキスによる、デコリン分解抑制の相乗効果の評価結果を図５に示す。
図５に示すように、オトギリソウエキスとゲンノショウコエキスの同時処置は、単独のデコリン分解抑制効果を足した理論値と比較して、デコリン分解抑制効果を増強させた。また、オトギリソウエキスとゲンノショウコエキスの添加量の比が１：４のときに、より高い相乗効果が得られた。 [Synergistic effect of Geranium thunbergii extract and Hypericum erectum extract in suppressing decorin decomposition]
FIG. 5 shows the evaluation results of the synergistic effect of suppressing the decomposition of decorin by Geranium thunbergii extract and Hypericum erectum extract.
As shown in FIG. 5, the simultaneous treatment of Hypericum erectum extract and Geranium thunbergii extract enhanced the decorin degradation inhibitory effect as compared with the theoretical value obtained by adding the decorin degradation inhibitory effect alone. Further, when the ratio of the addition amount of Hypericum erectum extract and Geranium thunbergii extract was 1: 4, a higher synergistic effect was obtained.

〔ゲンノショウコエキスとテンチャエキスのデコリン分解抑制における相乗効果〕
ゲンノショウコエキスとテンチャエキスによる、デコリン分解抑制の相乗効果の評価結果を図６に示す。
図６に示すように、テンチャエキスとゲンノショウコエキスの同時処置は、単独のデコリン分解抑制効果を足した理論値と比較して、デコリン分解抑制効果を増強させた。また、テンチャエキスとゲンノショウコエキスの添加量の比が１：４のときに、より高い相乗効果が得られた。 [Synergistic effect of Geranium thunbergii extract and Tencha extract in suppressing decorin degradation]
FIG. 6 shows the evaluation results of the synergistic effect of suppressing decorin degradation by Geranium thunbergii extract and Tencha extract.
As shown in FIG. 6, the simultaneous treatment of the tencha extract and the geranium thunbergii extract enhanced the decorin degradation inhibitory effect as compared with the theoretical value obtained by adding the decorin degradation inhibitory effect alone. Further, when the ratio of the addition amount of the tencha extract and the geranium thunbergii extract was 1: 4, a higher synergistic effect was obtained.

本発明は化粧料組成物、医薬組成物等に利用可能である。 The present invention can be used for cosmetic compositions, pharmaceutical compositions and the like.

Claims (24)

ゲンノショウコエキスを有効成分として含有する、グランザイムＢ抑制剤。 Granzyme B inhibitor containing Geranium thunbergii extract as an active ingredient. ゲンノショウコエキスを１〜１００μｇ／ｍＬ含有する、請求項１に記載のグランザイムＢ抑制剤。 The Granzyme B inhibitor according to claim 1, which contains 1 to 100 μg / mL of Geranium thunbergii extract. ゲンノショウコエキスを１．８〜５５μｇ／ｍＬ含有する、請求項２に記載のグランザイムＢ抑制剤。 The Granzyme B inhibitor according to claim 2, which contains 1.8 to 55 μg / mL of Geranium thunbergii extract. ゲンノショウコエキスを１８〜５５μｇ／ｍＬ含有する、請求項３に記載のグランザイムＢ抑制剤。 The Granzyme B inhibitor according to claim 3, which contains 18 to 55 μg / mL of Geranium thunbergii extract. グランザイムＢ抑制作用を有する１種以上の植物エキスをさらに含有する、請求項１から４のいずれか一項に記載のグランザイムＢ抑制剤。 The granzyme B inhibitor according to any one of claims 1 to 4, further comprising one or more plant extracts having a granzyme B inhibitory effect. 前記植物エキス及び前記ゲンノショウコエキスを１：１〜１：４の質量比で含有する、請求項５に記載のグランザイムＢ抑制剤。 The Granzyme B inhibitor according to claim 5, which contains the plant extract and the geranium thunbergii extract in a mass ratio of 1: 1 to 1: 4. 前記植物エキスが、ザクロ果皮エキス、イエローヒマラヤンラズベリー根エキス、ユーカリエキス、ワレモコウエキス、イロハモミジ葉エキス、シモツケソウエキス、ハマメリスエキス、バラエキス、アボガドエキス、テンチャエキス、メマツヨイグサ抽出液、ウーロン茶エキス、カシア樹皮エキス、ユキノシタエキス、メリッサエキス、トルメンチラエキス、ダマスクバラ花エキス、ヨモギエキス、ヒキオコシエキス、コンフリーエキス、オトギリソウエキス、セイヨウハッカエキス、ブドウ葉エキス、シナノキエキス、エイジツエキス、ビワ葉エキス、海藻エキス、シャクヤクエキス、ドクダミエキス、サンザシエキス、シソエキス、ラベンダーエキス、ノバラエキス、アセンヤクエキス、サガラメエキス、ボタンエキス、アスパラサスリネアリスエキス、カンゾウ抽出末及びクララエキスからなる群から選択される、請求項５又は６に記載のグランザイムＢ抑制剤。 The plant extracts are pomegranate peel extract, yellow Himalayan raspberry root extract, eucalyptus extract, crackle extract, sardine leaf extract, shimotsukesou extract, hamamelis extract, rose extract, avocado extract, tencha extract, mematsuyoigusa extract, oolong tea extract, cassia bark extract, Yukinoshita extract, Melissa extract, Tormentilla extract, Damask rose flower extract, Yomogi extract, Hikiokoshi extract, Confree extract, Otogirisou extract, Seiyouhakka extract, Grape leaf extract, Shinanoki extract, Agetsu extract, Biwa leaf extract, Seaweed extract, Claim 5 or 6 selected from the group consisting of Shakuyaku extract, Dokudami extract, Sanzashi extract, Perilla extract, Lavender extract, Novara extract, Asenyaku extract, Sagarame extract, Button extract, Asparasas linearis extract, Kanzo extract powder and Clara extract. Granzyme B inhibitor according to. 前記植物エキスの少なくとも１種が、オトギリソウエキス及びテンチャエキスからなる群から選択される、請求項７に記載のグランザイムＢ抑制剤。 The Granzyme B inhibitor according to claim 7, wherein at least one of the plant extracts is selected from the group consisting of Hypericum erectum extract and Tencha extract. グランザイムＢ抑制が、グランザイムＢ遺伝子の発現抑制である、請求項１から８のいずれか一項に記載のグランザイムＢ抑制剤。 The granzyme B inhibitor according to any one of claims 1 to 8, wherein the granzyme B inhibition is the inhibition of the expression of the granzyme B gene. グランザイムＢ抑制が、グランザイムＢの活性阻害である、請求項１から８のいずれか一項に記載のグランザイムＢ抑制剤。 The granzyme B inhibitor according to any one of claims 1 to 8, wherein the inhibition of granzyme B is an inhibition of the activity of granzyme B. グランザイムＢ抑制が、グランザイムＢ遺伝子の発現抑制及びグランザイムＢの活性阻害である、請求項１から８のいずれか一項に記載のグランザイムＢ抑制剤。 The granzyme B inhibitor according to any one of claims 1 to 8, wherein the inhibition of granzyme B is an inhibition of the expression of the granzyme B gene and an inhibition of the activity of granzyme B. 請求項１から１１のいずれか一項に記載のグランザイムＢ抑制剤のみからなる、化粧料組成物。 A cosmetic composition comprising only the Granzyme B inhibitor according to any one of claims 1 to 11. ゲンノショウコエキス及びオトギリソウエキスを有効成分として含有する、コラーゲン分解抑制剤。 A collagen decomposition inhibitor containing Geranium thunbergii extract and Hypericum erectum extract as active ingredients. 前記オトギリソウエキス及び前記ゲンノショウコエキスを１：１〜１：４の質量比で含有する、請求項１３に記載のコラーゲン分解抑制剤。 The collagen decomposition inhibitor according to claim 13, which contains the Hypericum erectum extract and the Geranium thunbergii extract in a mass ratio of 1: 1 to 1: 4. ゲンノショウコエキス及びテンチャエキスを有効成分として含有する、コラーゲン分解抑制剤。 A collagen decomposition inhibitor containing Geranium thunbergii extract and Tencha extract as active ingredients. 前記テンチャエキス及び前記ゲンノショウコエキスを１：１〜１：４の質量比で含有する、請求項１５に記載のコラーゲン分解抑制剤。 The collagen decomposition inhibitor according to claim 15, which contains the tencha extract and the geranium thunbergii extract in a mass ratio of 1: 1 to 1: 4. ゲンノショウコエキスを１〜１００μｇ／ｍＬ含有する、請求項１３〜１６のいずれか一項に記載のコラーゲン分解抑制剤。 The collagen decomposition inhibitor according to any one of claims 13 to 16, which contains 1 to 100 μg / mL of Geranium thunbergii extract. ゲンノショウコエキスを１．８〜５５μｇ／ｍＬ含有する、請求項１７に記載のコラーゲン分解抑制剤。 The collagen decomposition inhibitor according to claim 17, which contains 1.8 to 55 μg / mL of Geranium thunbergii extract. ゲンノショウコエキスを１８〜５５μｇ／ｍＬ含有する、請求項１８に記載のコラーゲン分解抑制剤。 The collagen decomposition inhibitor according to claim 18, which contains 18 to 55 μg / mL of Geranium thunbergii extract. コラーゲン分解抑制作用を有する１種以上の植物エキスをさらに含有する、請求項１３から１９のいずれか一項に記載のコラーゲン分解抑制剤。 The collagen degradation inhibitor according to any one of claims 13 to 19 , further comprising one or more plant extracts having a collagen degradation inhibitory action. 前記植物エキスが、ザクロ果皮エキス、イエローヒマラヤンラズベリー根エキス、ユーカリエキス、ワレモコウエキス、イロハモミジ葉エキス、シモツケソウエキス、ハマメリスエキス、バラエキス、アボガドエキス、メマツヨイグサ抽出液、ウーロン茶エキス、カシア樹皮エキス、ユキノシタエキス、メリッサエキス、トルメンチラエキス、ダマスクバラ花エキス、ヨモギエキス、ヒキオコシエキス、コンフリーエキス、セイヨウハッカエキス、ブドウ葉エキス、シナノキエキス、エイジツエキス、ビワ葉エキス、海藻エキス、シャクヤクエキス、ドクダミエキス、サンザシエキス、シソエキス、ラベンダーエキス、ノバラエキス、アセンヤクエキス、サガラメエキス、ボタンエキス、アスパラサスリネアリスエキス、カンゾウ抽出末及びクララエキスからなる群から選択される、請求項２０に記載のコラーゲン分解抑制剤。 The plant extracts are pomegranate peel extract, yellow Himalayan raspberry root extract, eucalyptus extract, crackle extract, Irohamomiji leaf extract, shimotsukesou extract, hamamelis extract, rose extract, avocado extract , mematsuyoigusa extract, oolong tea extract, cassia bark extract, yukinoshita extract, Melissa extract, take Men Chira extract, damask rose flower extract, mugwort extract, cause extract, comfrey extract, peppermint extract, grape leaf extract, linden extract, rose fruit extract, loquat leaf extract, seaweed extract, peony root extract, Houttuynia cordata extract, The collagen decomposition inhibitor according to claim 20 , which is selected from the group consisting of Sanzashi extract, Perilla extract, Lavender extract, Novara extract, Asenyaku extract, Sagarame extract, Button extract, Asparasus linearis extract, Kanzo extract powder and Clara extract. コラーゲン分解抑制が、グランザイムＢ遺伝子の発現抑制である、請求項１３から２１のいずれか一項に記載のコラーゲン分解抑制剤。 The collagen degradation inhibitor according to any one of claims 13 to 21 , wherein the inhibition of collagen degradation is the inhibition of the expression of the Granzyme B gene. コラーゲン分解抑制が、グランザイムＢの活性阻害である、請求項１３から２１のいずれか一項に記載のコラーゲン分解抑制剤。 The collagen degradation inhibitor according to any one of claims 13 to 21 , wherein the inhibition of collagen degradation is an inhibition of the activity of Granzyme B. コラーゲン分解抑制が、グランザイムＢ遺伝子の発現抑制及びグランザイムＢの活性阻害である、請求項１３から２１のいずれか一項に記載のコラーゲン分解抑制剤。 The collagen degradation inhibitor according to any one of claims 13 to 21 , wherein the inhibition of collagen degradation is the inhibition of the expression of the Granzyme B gene and the inhibition of the activity of Granzyme B.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102510213B1 (en) * 2022-09-30 2023-03-15 아람휴비스 주식회사 The adhesive transparent wound covering material composition containing fumaria officinalis, pomegranate shell extract and comfrey leaf extract

Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS62148426A (en) * 1985-12-23 1987-07-02 Rooto Seiyaku Kk Remedy for periodontosis
JPH11315008A (en) * 1998-03-03 1999-11-16 Shiseido Co Ltd Antiaging agent
JP2003192565A (en) * 2001-12-27 2003-07-09 Nonogawa Shoji Kk Cosmetic
JP2006131558A (en) * 2004-11-05 2006-05-25 Nippon Menaade Keshohin Kk Collagen formation enhancer
WO2007046353A1 (en) * 2005-10-18 2007-04-26 Mercian Corporation Fibroblast activator, method for activation of fibroblast, collagen synthesis promoter, method for promotion of collagen synthesis, skin aging-preventing agent, and method for prevention of aging of the skin
WO2008026295A1 (en) * 2006-09-01 2008-03-06 Toyo Shinyaku Co., Ltd. Supporter
JP2009221110A (en) * 2008-03-13 2009-10-01 Shiseido Co Ltd Basement membrane stabilizer
JP2010241714A (en) * 2009-04-03 2010-10-28 Kao Corp Aggrecan decomposition inhibitor
JP2017078061A (en) * 2015-10-21 2017-04-27 大正製薬株式会社 Scalp agents
KR20200020404A (en) * 2018-08-17 2020-02-26 서울대학교산학협력단 Composition for inhibition of granzyme b comprising plant extract or compound therefrom
JP2020515511A (en) * 2016-12-23 2020-05-28 ナチュレクス セーアー Composition

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4139545B2 (en) * 2000-03-17 2008-08-27 花王株式会社 Skin cosmetics
EP3143983A1 (en) * 2003-02-13 2017-03-22 Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo External dermatological formulation comprising saccharide derivative of alpha, alpha-trehalose
CN101528190B (en) * 2006-09-26 2013-03-20 狮王株式会社 Shampoo composition
CN101653463B (en) * 2009-09-18 2011-09-28 延边大学 Extraction and purification method of tannin components in Geranium wilfordii
JP7232642B2 (en) * 2018-12-28 2023-03-03 ポーラ化成工業株式会社 Anti-aging method and anti-aging agent

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS62148426A (en) * 1985-12-23 1987-07-02 Rooto Seiyaku Kk Remedy for periodontosis
JPH11315008A (en) * 1998-03-03 1999-11-16 Shiseido Co Ltd Antiaging agent
JP2003192565A (en) * 2001-12-27 2003-07-09 Nonogawa Shoji Kk Cosmetic
JP2006131558A (en) * 2004-11-05 2006-05-25 Nippon Menaade Keshohin Kk Collagen formation enhancer
WO2007046353A1 (en) * 2005-10-18 2007-04-26 Mercian Corporation Fibroblast activator, method for activation of fibroblast, collagen synthesis promoter, method for promotion of collagen synthesis, skin aging-preventing agent, and method for prevention of aging of the skin
WO2008026295A1 (en) * 2006-09-01 2008-03-06 Toyo Shinyaku Co., Ltd. Supporter
JP2009221110A (en) * 2008-03-13 2009-10-01 Shiseido Co Ltd Basement membrane stabilizer
JP2010241714A (en) * 2009-04-03 2010-10-28 Kao Corp Aggrecan decomposition inhibitor
JP2017078061A (en) * 2015-10-21 2017-04-27 大正製薬株式会社 Scalp agents
JP2020515511A (en) * 2016-12-23 2020-05-28 ナチュレクス セーアー Composition
KR20200020404A (en) * 2018-08-17 2020-02-26 서울대학교산학협력단 Composition for inhibition of granzyme b comprising plant extract or compound therefrom

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102510213B1 (en) * 2022-09-30 2023-03-15 아람휴비스 주식회사 The adhesive transparent wound covering material composition containing fumaria officinalis, pomegranate shell extract and comfrey leaf extract

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