JP2022125934A - Yeast extract having cell migration ability - Google Patents
Yeast extract having cell migration ability Download PDFInfo
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- JP2022125934A JP2022125934A JP2021023783A JP2021023783A JP2022125934A JP 2022125934 A JP2022125934 A JP 2022125934A JP 2021023783 A JP2021023783 A JP 2021023783A JP 2021023783 A JP2021023783 A JP 2021023783A JP 2022125934 A JP2022125934 A JP 2022125934A
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本発明は酵母由来の食物繊維を有効成分として含有する、肌荒れ改善効果に優れた化粧料組成物および医薬組成物を提供するものである。 The present invention provides a cosmetic composition and a pharmaceutical composition that contain yeast-derived dietary fiber as an active ingredient and are excellent in the effect of improving rough skin.
健康的な肌を保つためや手入れを目的に、化粧水や乳液などの化粧品や外用薬が用いられる。 Cosmetics such as lotions and milky lotions and topical medicines are used to maintain and care for healthy skin.
紫外線などの外的要因によって荒れた肌を改善する機能として、細胞遊走能があり例えば合成ペプチドなどが知られている（特許文献１）。 Synthetic peptides, for example, are known to have cell migration ability as a function to improve rough skin caused by external factors such as ultraviolet rays (Patent Document 1).
化粧品全般においては、近年においては安全性の観点からも合成物ではなく天然由来のものが求められており、例えば細胞分化を訴求効果とするマンネンタケ胞子抽出物などを有効成分として含むもの(特許文献２)やヒアルロニダーゼ阻害を訴求効果とするバラ科植物抽出物を有効成分として含むもの(特許文献３）が知られている。また、メラニン産生抑制を訴求効果とするトルラ酵母由来グルコシルセラミンドなどがある（特許文献４）。
しかしながら、酵母由来のもので肌荒れを改善させるものは知られていなかった。
In cosmetics in general, in recent years, from the viewpoint of safety, there is a demand for naturally derived products rather than synthetic products. 2) and those containing a Rosaceae plant extract as an active ingredient (Patent Document 3), which have an appealing effect of inhibiting hyaluronidase. In addition, there is a torula yeast-derived glucosylceramide that has an appealing effect of suppressing melanin production (Patent Document 4).
However, yeast-derived substances that improve rough skin have not been known.
本発明は酵母由来の食物繊維を有効成分として含有する、肌荒れ改善効果に優れた化粧料組成物および医薬組成物を提供するものである。 The present invention provides a cosmetic composition and a pharmaceutical composition that contain yeast-derived dietary fiber as an active ingredient and are excellent in the effect of improving rough skin.
本発明者らは鋭意検討の結果正常ヒト線維芽細胞に対して、酵母由来の食物繊維を含む抽出物に細胞遊走能を有し、肌荒れ改善に効果があることを見出した。 As a result of intensive studies, the present inventors have found that an extract containing yeast-derived dietary fiber has cell migration ability and is effective in improving rough skin against normal human fibroblasts.
すなわち、本発明は
(１) 酵母抽出物を含有する肌荒れ改善用組成物
(２) 食物繊維を含有する酵母抽出物を含む肌荒れ改善用組成物
(３) 前記組成物が化粧料組成物または医薬組成物である、（１）または（２）記載の組成物
に係るものである。
That is, the present invention
(1) Composition for improving rough skin containing yeast extract
(2) A composition for improving rough skin containing a yeast extract containing dietary fiber
(3) The composition according to (1) or (2), wherein the composition is a cosmetic composition or a pharmaceutical composition.
本発明の酵母由来の食物繊維を含む抽出物は、食品として知られている酵母から抽出できるため、比較的容易かつ安価に生産でき、肌荒れ改善等の皮膚の状態を改善する組成物として安全に使用することが可能である。 The extract containing yeast-derived dietary fiber of the present invention can be extracted from yeast known as food, so it can be produced relatively easily and inexpensively, and can be safely used as a composition that improves skin conditions such as rough skin. It is possible to use
本発明の食物繊維を含む抽出物の製造に用いられる酵母は、一般的に用いられている酵母であれば使用することができる。具体的には、パン酵母、ビール酵母（サッカロマイセス・セルビシエ）トルラ酵母（キャンディダ・ユティリス）などを挙げることができる。この中でも、特にトルラ酵母が望ましい。酵母の培養方法も、食品等の産業で一般的に採用される方法で、培養でき、本発明では制限がない。 Any commonly used yeast can be used for the production of the dietary fiber-containing extract of the present invention. Specific examples include baker's yeast, brewer's yeast (Saccharomyces cerevisiae), torula yeast (Candida utilis), and the like. Among these, torula yeast is particularly desirable. Yeast can also be cultured by any method generally employed in industries such as foods, and there is no limitation in the present invention.
本発明の食物繊維を含有する酵母菌体から抽出することで得られる。酵母菌体としては可溶性エキス成分を分離除去した後の菌体残渣を用いるのが好ましい。可溶性エキス成分の除去方法は、食品等に用いられる酵母エキスの一般的な抽出方法で、可溶性分を除去すればよく、特に制限はなく、本発明では適用できる。通常は、酵母に熱水、アルカリ性溶液、自己消化、機械的破砕、細胞壁溶解酵素、蛋白質分解酵素、リボヌクレアーゼ、またはデアミナーゼのいずれか一つ以上を用いてエキスを抽出し、可溶性成分を除去する。なお、可溶性成分を除去した菌体残渣として、一般に販売されているものを使用することもできる。市販品としては、例えば、三菱商事ライフサイエンス社製の「KR酵母」、「GL酵母」などを用いることもできる。 It is obtained by extraction from yeast cells containing the dietary fiber of the present invention. As the yeast cells, it is preferable to use the cell residue after separating and removing the soluble extract components. The method for removing soluble extract components is a general method for extracting yeast extracts used in foods and the like, and the soluble components can be removed without particular limitation, and can be applied in the present invention. Usually, yeast is subjected to one or more of hot water, alkaline solution, autolysis, mechanical disruption, cell wall lytic enzyme, proteolytic enzyme, ribonuclease, or deaminase to extract the extract and remove soluble components. In addition, the thing generally sold can also be used as a bacterial cell residue which removed the soluble component. As commercially available products, for example, "KR Yeast" and "GL Yeast" manufactured by Mitsubishi Corporation Life Sciences can be used.
上記の菌体残渣にグルカナーゼを作用することで肌荒れ改善効果のある酵母抽出物を得ることができる。グルカナーゼは特に制限はないが、トルラ酵母を用いる場合は、長瀬ケムテックス製の精製グルカナーゼ製剤「デナチームGEL」が好ましい。例えば、長瀬ケムテックス製のグルカナーゼ製剤「デナチームGEL-L1」の場合、酵母抽出物をｐＨ４～７、望ましくはｐＨ５～６に調整し、25～75℃、望ましくは30～60℃で、酵母菌体重量に対して酵素を０．０５～２％、望ましくは０．１～１％添加し、１～30時間、望ましくは10～1５時間反応させる。こうして得られる本願発明の酵母抽出物は食物繊維を含有するものであり、食物繊維を５重量％以上、望ましくは１５重量％以上、さらに望ましくは５０重量％以上含有するものである。 By acting glucanase on the bacterial cell residue, it is possible to obtain a yeast extract that is effective in improving rough skin. The glucanase is not particularly limited, but when torula yeast is used, a purified glucanase preparation "Denazyme GEL" manufactured by Nagase Chemtex is preferred. For example, in the case of Nagase Chemtex's glucanase preparation "Denazyme GEL-L1", the yeast extract is adjusted to pH 4 to 7, preferably pH 5 to 6, and heated to 25 to 75 ° C., preferably 30 to 60 ° C. Yeast cells 0.05 to 2%, preferably 0.1 to 1%, of the enzyme is added to the weight and allowed to react for 1 to 30 hours, preferably 10 to 15 hours. The yeast extract of the present invention thus obtained contains dietary fiber, preferably 5% by weight or more, preferably 15% by weight or more, more preferably 50% by weight or more.
本発明の肌荒れ改善における有効成分は酵母抽出物であるため、酵母抽出物をそのまま肌荒れ改善効果のある組成物として使用することもでき、また他の素材との混合物にして使用してもよい。そのまま、食品や医薬品として使用することができる。食品として使用する場合は、機能性食品、栄養補助食品、サプリメント等として摂取でき、化粧品、塗布剤としても使用できる。 Since the active ingredient for improving rough skin according to the present invention is a yeast extract, the yeast extract can be used as it is as a composition effective in improving rough skin, or it can be used as a mixture with other materials. It can be used as a food or a medicine as it is. When used as a food, it can be ingested as a functional food, dietary supplement, supplement, etc., and can also be used as a cosmetic or coating agent.
また、本発明の組成物は、他の食品素材や医薬品素材、化粧品素材と混合して使用しても良い。例えば、賦形剤、希釈剤となるデキストリン、マルチトール、ソルビトール、デンプンなどである。化粧品や塗布剤等として使用する場合、その形態は特に限定されるものではなく、クリーム状、乳液状、ローション状、軟膏状、ジェル状、スプレー、ムース状、油中水型エマルション、水中油型エマルション、固形状、シート状、パウダー状等々に適用が可能である。 Moreover, the composition of the present invention may be used by being mixed with other food materials, pharmaceutical materials, or cosmetic materials. For example, excipients and diluents such as dextrin, maltitol, sorbitol and starch. When it is used as a cosmetic or a topical agent, its form is not particularly limited and includes cream, milky lotion, lotion, ointment, gel, spray, mousse, water-in-oil emulsion, and oil-in-water. It can be applied in emulsion, solid form, sheet form, powder form, and the like.
本製品を投与量、塗布量は、対象となる皮膚の状態、投与対象の年齢、性別、体重、症状の程度などに応じて適宜決定することができる。 The dose and application amount of this product can be appropriately determined according to the target skin condition, age, sex, weight, severity of symptoms, etc. of the target.
以下に実施例を挙げて本発明をさらに詳細に説明するが、本発明はこれら実施例に限定されるものではない。 EXAMPLES The present invention will be described in more detail with reference to examples below, but the present invention is not limited to these examples.
（本願発明の抽出物の製造）
「ＫＲ酵母」（三菱商事ライフサイエンス社製）１０ｇを固形分 １０％（１０ｇ／１００ ｍＬ）となるように調整した。調整後６０℃、ｐＨ６．５にて、デナチームＧＥＬ―１を０．０５ｇとなるよう添加した。添加後撹拌しながら、３時間反応し、反応後、それぞれを遠心分離し、上清を食物繊維を含有する抽出物として回収した。回収後、噴霧乾燥し、乾燥物を２．５ｇ得た。得られた乾燥物の食物繊維を酵素―ＨＰＬＣ法で測定したところ、５１％であった。
(Production of the extract of the present invention)
10 g of "KR Yeast" (manufactured by Mitsubishi Corporation Life Sciences) was adjusted to a solid content of 10% (10 g/100 mL). Denatzyme GEL-1 was added to 0.05 g at 60° C. and pH 6.5 after adjustment. After the addition, the mixture was reacted for 3 hours with stirring. After the reaction, each was centrifuged, and the supernatant was recovered as an extract containing dietary fiber. After collection, it was spray-dried to obtain 2.5 g of a dried product. The dietary fiber content of the dried product obtained was measured by an enzyme-HPLC method and found to be 51%.
正常ヒト線維芽細胞を２４ウェルプレートに播種（１．５×１０５ｃｅｌｌｓ／ｗｅｌｌ）し、２４時間前培養した（５％ＣＯ２、３７℃）。培養液は、１０％牛胎児血清を含むＭＥＭ－α、ｎｕｃｌｅｏｓｉｄｅｓ培地を使用した。その後、スクラッチスティック（ＡＧＣテクノグラス製）を用いてウェルの縦方向に幅３ｍｍの亀裂を入れ、ＰＢＳ（－）でウェルを１回洗浄した後、実施例１で得られた抽出物の終濃度が１０，５０，１００μｇ／ｍＬとなるように溶解した牛胎児血清を含まないＭＥＭ－α、ｎｕｃｌｅｏｓｉｄｅ培地に交換し、さらに２４時間培養した。抽出物は、ＤＭＳＯの終濃度が０．５％となるように溶解して用いた。
培養後の細胞遊走能は、生細胞をクリスタルバイオレット染色することで評価した。上述のように２４時間培養した後、ＰＢＳ（－）でウェルを１回洗浄し、４％パラホルムアルデヒド溶液を加えて２０分間静置した。続いてＰＢＳ（－）でウェルを１回洗浄し、０．１％クリスタルバイオレット含有１％酢酸溶液を加えて１５分間静置した後、ＰＢＳ（－）でウェルを２回洗浄し、顕微鏡観察を行った。亀裂部位の同一面積当たりの細胞数を細胞遊走能として評価した。
Normal human fibroblasts were seeded in a 24-well plate (1.5×10 5 cells/well) and precultured for 24 hours (5% CO 2 , 37° C.). The culture medium used was MEM-α, nucleosides medium containing 10% fetal bovine serum. After that, using a scratch stick (manufactured by AGC Techno Glass), a crack of 3 mm in width was made in the vertical direction of the well, and after washing the well once with PBS (-), the final concentration of the extract obtained in Example 1 was The medium was replaced with a MEM-α, nucleoside medium containing no fetal bovine serum dissolved so that the concentration was 10, 50 or 100 μg/mL, and cultured for an additional 24 hours. The extract was used after being dissolved in DMSO to a final concentration of 0.5%.
Cell migration ability after culture was evaluated by crystal violet staining of living cells. After culturing for 24 hours as described above, the wells were washed once with PBS(-), 4% paraformaldehyde solution was added, and the cells were allowed to stand for 20 minutes. Subsequently, the wells were washed once with PBS(-), a 1% acetic acid solution containing 0.1% crystal violet was added and allowed to stand for 15 minutes, then the wells were washed twice with PBS(-) and observed under a microscope. gone. The number of cells per the same area of the crack site was evaluated as the cell migration ability.
実施例１で得られた抽出物を用いないこと以外は実施例２と同様に行った。 The procedure of Example 2 was repeated except that the extract obtained in Example 1 was not used.
実施例１と比較例１について、亀裂部位の同一面積当たりの細胞数を比較した。実施例１及び比較例１の顕微鏡写真を図２に、比較例１の細胞数を１００％とした場合の細胞遊走能を表１に記載した。実施例１では比較例１と比べて濃度依存的に細胞遊走能が向上していたことから、肌荒れの改善に有効であることが分かった。 A comparison was made between Example 1 and Comparative Example 1 in terms of the number of cells per the same area of the cracked site. Microphotographs of Example 1 and Comparative Example 1 are shown in FIG. In Example 1, the cell migration ability was improved in a concentration-dependent manner as compared with Comparative Example 1, and thus it was found to be effective in alleviating rough skin.
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