JP3784704B2 - Cosmetics - Google Patents
Cosmetics Download PDFInfo
- Publication number
- JP3784704B2 JP3784704B2 JP2001368152A JP2001368152A JP3784704B2 JP 3784704 B2 JP3784704 B2 JP 3784704B2 JP 2001368152 A JP2001368152 A JP 2001368152A JP 2001368152 A JP2001368152 A JP 2001368152A JP 3784704 B2 JP3784704 B2 JP 3784704B2
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- JP
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- Prior art keywords
- collagen
- loofah
- hybrid cell
- molecular weight
- cell extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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- 239000002537 cosmetic Substances 0.000 title claims description 26
- 102000008186 Collagen Human genes 0.000 claims description 64
- 108010035532 Collagen Proteins 0.000 claims description 64
- 229920001436 collagen Polymers 0.000 claims description 62
- 210000004754 hybrid cell Anatomy 0.000 claims description 29
- 244000280244 Luffa acutangula Species 0.000 claims description 25
- 235000009814 Luffa aegyptiaca Nutrition 0.000 claims description 25
- 239000000284 extract Substances 0.000 claims description 22
- 230000037319 collagen production Effects 0.000 claims description 13
- 230000003712 anti-aging effect Effects 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 241000251468 Actinopterygii Species 0.000 claims description 6
- 244000267823 Hydrangea macrophylla Species 0.000 claims description 3
- 235000014486 Hydrangea macrophylla Nutrition 0.000 claims description 3
- 238000013329 compounding Methods 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 description 15
- 238000012360 testing method Methods 0.000 description 15
- 239000007788 liquid Substances 0.000 description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 11
- 238000004519 manufacturing process Methods 0.000 description 10
- 239000000243 solution Substances 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 7
- 239000000686 essence Substances 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 230000001737 promoting effect Effects 0.000 description 6
- 210000003491 skin Anatomy 0.000 description 6
- 241000972773 Aulopiformes Species 0.000 description 5
- 235000019515 salmon Nutrition 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 230000037394 skin elasticity Effects 0.000 description 4
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 240000006509 Gynostemma pentaphyllum Species 0.000 description 3
- 235000002956 Gynostemma pentaphyllum Nutrition 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 3
- BTANRVKWQNVYAZ-UHFFFAOYSA-N butan-2-ol Chemical compound CCC(C)O BTANRVKWQNVYAZ-UHFFFAOYSA-N 0.000 description 3
- 230000002500 effect on skin Effects 0.000 description 3
- 235000019688 fish Nutrition 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000037303 wrinkles Effects 0.000 description 3
- 229940058015 1,3-butylene glycol Drugs 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 102000029816 Collagenase Human genes 0.000 description 2
- 108060005980 Collagenase Proteins 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 206010020649 Hyperkeratosis Diseases 0.000 description 2
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 2
- 102000057297 Pepsin A Human genes 0.000 description 2
- 108090000284 Pepsin A Proteins 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- 150000001298 alcohols Chemical class 0.000 description 2
- 235000019437 butane-1,3-diol Nutrition 0.000 description 2
- 229960002424 collagenase Drugs 0.000 description 2
- 238000004042 decolorization Methods 0.000 description 2
- 238000004332 deodorization Methods 0.000 description 2
- 210000004207 dermis Anatomy 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 239000000469 ethanolic extract Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 229930195733 hydrocarbon Natural products 0.000 description 2
- 150000002430 hydrocarbons Chemical class 0.000 description 2
- 239000006210 lotion Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000003020 moisturizing effect Effects 0.000 description 2
- -1 pH adjusters Substances 0.000 description 2
- 229940111202 pepsin Drugs 0.000 description 2
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical compound OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000005185 salting out Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 230000009759 skin aging Effects 0.000 description 2
- 210000001626 skin fibroblast Anatomy 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 2
- 150000005846 sugar alcohols Polymers 0.000 description 2
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000534672 Hiodon alosoides Species 0.000 description 1
- 239000002211 L-ascorbic acid Substances 0.000 description 1
- 235000000069 L-ascorbic acid Nutrition 0.000 description 1
- 239000004909 Moisturizer Substances 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 102000012479 Serine Proteases Human genes 0.000 description 1
- 108010022999 Serine Proteases Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000006096 absorbing agent Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000012670 alkaline solution Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000003788 bath preparation Substances 0.000 description 1
- AGSPXMVUFBBBMO-UHFFFAOYSA-N beta-aminopropionitrile Chemical compound NCCC#N AGSPXMVUFBBBMO-UHFFFAOYSA-N 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000012459 cleaning agent Substances 0.000 description 1
- 230000011382 collagen catabolic process Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- POLCUAVZOMRGSN-UHFFFAOYSA-N dipropyl ether Chemical compound CCCOCCC POLCUAVZOMRGSN-UHFFFAOYSA-N 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000003349 gelling agent Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 235000013772 propylene glycol Nutrition 0.000 description 1
- 238000007665 sagging Methods 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 239000001384 succinic acid Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
- Cosmetics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Description
【0001】
【発明の属する技術分野】
本発明は、マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物を配合したコラーゲン生成促進剤および抗老化用化粧料に関するものである。
【0002】
【従来の技術】
皮膚の老化現象のひとつとして、シワの発生や弾力性の低下があげられるが、これには真皮構成成分であるコラーゲンの減少が大きく影響することが知られている。そのため、コラーゲンを配合した数多くの化粧料やコラーゲン生成促進物質を配合した化粧料が開発されている。
【0003】
【発明が解決しようとする課題】
しかし、コラーゲンを配合した従来の化粧料は一時的な保湿効果はあるものの、真皮中のコラーゲン量を根本的に改善するものではなかった。また、コラーゲン生成促進物質を配合した化粧料の効果も十分ではなく、より効果のあるものが望まれていた。そこで、コラーゲン生成促進剤を組み合わせて化粧料に配合することにより、真皮コラーゲン量の減少をより効果的に抑制し、シワの発生や弾力性の低下を抑制できることを見出し、本発明に至ったものである。
【0004】
【課題を解決するための手段】
本発明は、マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物を配合することを特徴とする化粧料である。本発明でいうマリンコラーゲンは、魚(例えば、鮭、金目鯛、平目、鱈、鰯、鮪等)の皮、浮袋等から得られるコラーゲンである。すなわち、本願明細書に記載されているマリンコラーゲンは魚由来のコラーゲンを示している。最近ではマリンコラーゲンは保湿剤として使用されるようになってきているが、従来多用されているウシ由来のコラーゲンに比べ、マリンコラーゲンはほとんど研究されていなかった。今回、マリンコラーゲンにコラーゲン生成促進効果があることを見出した。さらに、加水分解による低分子のマリンコラーゲンにより高い効果のあることを見出した。また本発明は、コハク化コラーゲンやフタル化コラーゲン等の修飾したマリンコラーゲンも含む。
【0005】
また、ヘチマとアマチャヅルの雑種細胞抽出物には、セリンプロテアーゼ阻害効果によってコラーゲンの分解を抑制し、肌の老化を防ぐことは知られている(特開平11−124325)が、コラーゲン生成促進効果があることは全く知られていない。
【0006】
マリンコラーゲンにコラーゲン生成促進効果のあることを見出し、さらに効果を高めるために研究した結果、マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物を併用することにより、特に顕著な効果を発揮することを見出した。さらに、平均分子量200〜1,500の低分子マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物を併用することにより、より高い効果を発揮することを見出した。
【0007】
本発明で用いるマリンコラーゲンの抽出・精製は、通常公知の方法を用いて行うことができる。具体的には、例えば魚皮を粉砕した後に水洗し、酸或いはアルカリ溶液による抽出、ペプシン、トリプシンやヒアロニダーゼ等の酵素により抽出し、塩析や透析の公知の精製手段を施して、平均分子量30万程度のコラーゲン溶液を得ることができる。このコラーゲン溶液は、抽出した溶液のまま用いても良く、必要に応じて、濃縮、希釈、濾過等の処理および活性炭等による脱色、脱臭処理をして用いても良い。
さらに、このコラーゲン溶液に通常公知の方法でコラゲナーゼを加えることによって、分子量200〜1,500程度のコラーゲンペプチドにすることが可能である。
平均分子量に関わらず、こうして得られたコラーゲン溶液にコハク酸やフタル酸等を反応させて化学修飾した、コハク化コラーゲンやフタル化コラーゲンなどのマリンコラーゲンを使用することもできる。
【0008】
こうして得られたマリンコラーゲンは、濃縮乾固、噴霧乾燥、凍結乾燥等の処理を行い、乾燥物として用いても良い。
【0009】
上記のマリンコラーゲンは、単独で用いても良いし、2種以上の混合物として用いても良い。
【0010】
ヘチマとアマチャヅルの雑種細胞抽出物は特開平01−44297の方法により作出した雑種細胞カルスを用いることができる。
【0011】
ヘチマとアマチャヅルの雑種細胞のカルスは特開平01−44297の方法により作出した雑種細胞カルス(F3、F4、F5、F6)を単独で用いても良いし、2種以上の混合物として用いても良い。
【0012】
ヘチマとアマチャヅルの雑種細胞を抽出する溶媒としては、水以外に、例えば、低級アルコール類(メタノール、エタノール、1−プロパノール、2−プロパノール、1−ブタノール、2−ブタノール等)、液状多価アルコール(グリセリン、プロピレングリコール、1,3−ブチレングリコール等)、ケトン類(アセトン、メチルエチルケトン等)、アセトニトリル、エステル類(酢酸エチル、酢酸ブチル等)、炭化水素類(ヘキサン、ヘプタン、流動パラフィン等)、エーテル類(エチルエーテル、プロピルエーテル、テトラヒドロフラン等)が挙げられる。好ましくは、水、低級アルコールおよび液状多価アルコールが良く、特に好ましくは、水、エタノール、プロピレングリコールおよび1,3−ブチレングリコールが良い。これらの溶媒は一種でも二種以上を混合して用いても良い。
【0013】
本発明に用いるマリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物をあわせた配合量は、乾固物に換算して本発明化粧料の全量中、0.00001〜10重量%の配合が良い。0.00001重量%未満では十分な効果は望めない。10重量%を越えて配合した場合、効果の増強はなく不経済である。また、添加の方法については、予め加えておいても、製造途中に添加してもよく、作業性を考えて適宜選択すれば良い。
【0014】
本発明の化粧料には、マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物の効果を損なわない範囲内で、通常の化粧料に用いられる成分である油脂類、ロウ類、炭化水素類、脂肪酸類、アルコール類、エステル類、界面活性剤、金属石鹸、pH調整剤、防腐剤、香料、保湿剤、粉体、紫外線吸収剤、増粘剤、色素、酸化防止剤、キレート剤等の成分を配合することができる。
【0015】
本発明の抗老化用化粧料には、化粧水、乳液、クリーム、美容液、パック、ゲル剤、洗浄剤、浴用剤、ファンデーション、口紅、アイシャドウ等が挙げられる。
【0016】
【実施例】
以下に、本発明を実施例により詳しく説明する。ただし、本発明の技術的範囲がこれらの製造例等によって限定されるものではない。実施例に示す配合量の部とは重量部を示し、%は重量%を示す。
【0017】
製造例1:鮭の皮由来のマリンコラーゲン
鮭の皮1kgを用い、通常公知の方法により得られたコラーゲン溶液を濃縮、希釈、濾過等の処理および活性炭等による脱色、脱臭処理を施し、抽出した溶液を凍結乾燥処理し、平均分子量30万のコラーゲン粉末140gを得た。
さらに、上記コラーゲン粉末に熱水を加えゼラチンに変性させ、通常公知の方法でコラゲナーゼを加えて加水分解したコラーゲンペプチド溶液を凍結乾燥処理し、低分子のコラーゲンペプチド粉末135gを得た。
【0018】
製造例2:ヘチマとアマチャヅルの雑種細胞50%エタノール抽出物
ヘチマとアマチャヅルの雑種細胞(F5)100gに1,000mLの50%エタノールを加え、常温で7日間抽出した後、ろ過し、その濾液を濃縮し、乾固して、ヘチマとアマチャヅルの雑種細胞50%エタノール抽出物7gを得た。
【0019】
実施例 美容液
マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物の配合量を変化させることによって、表1〜2に示す美容液1〜5、比較例1〜2をつくることができる。
【0020】
【表1】
【0021】
【表2】
【0022】
[製造方法]表1〜2とも、成分1〜7及び12と、成分8〜11をそれぞれ均一に溶解し、両者を混合して調整する。
【0023】
【発明の効果】
次に、本発明の効果を詳細に説明するため、実験例を挙げる。
【0024】
実験例1 コラーゲン生成促進作用
試験には、供試細胞としてヒト皮膚線維芽細胞(6歳男子由来、継代数6)を用い、培地としてEagle’s MEMを用いた。コラーゲン生成量の測定は、コラーゲンタンパク中への3H−プロリンの取り込みを指標として行った。96穴プレート(Falcon3072)にヒト皮膚線維芽細胞を25,000cells/well播種し、10%FCSを含むEagle’s MEMにて、37度、5%CO2条件下で4日間培養した。この間に細胞は、コンフルエントの状態となった。次に、本発明のマリンコラーゲン(製造例1;平均分子量100、平均分子量200、平均分子量500、平均分子量1,500、平均分子量3,000、平均分子量30万)、ヘチマとアマチャヅルの雑種細胞抽出物(製造例2)、マリンコラーゲン(製造例1;平均分子量100、平均分子量200、平均分子量500、平均分子量1,500、平均分子量3,000、平均分子量30万)+ヘチマとアマチャヅルの雑種細胞抽出物(製造例2)を各々最終濃度5μg/mlとなるように調整した無血清のEagle’s MEM(50μg/ml L−アスコルビン酸、50μg/ml β−アミノプロピオニトリル、18.5KBq 3H―プロリン含有)に交換し、24時間培養した。その後、直ちに各wellにペプシン溶液を加えインキュベーションした後、3H―プロリン標識されたコラーゲンを抽出した。塩析により精製した後、コラーゲンタンパク中の放射活性を液体シンチレーションカウンター(Beckman LS 5801)により測定した。各試料とも5wellずつ測定し、生成量は平均値±標準偏差で示した。また、試料無添加のものをコントロールとして用いた。
【0025】
【表3】
【0026】
表3に示したように、本発明のマリンコラーゲン+ヘチマとアマチャヅルの雑種細胞抽出物はヒト皮膚線維芽細胞のコラーゲン生成を有意に促進した。単なるマリンコラーゲン、ヘチマとアマチャヅルの雑種細胞抽出物単独よりマリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物を併用することにより効果の向上が見られた。また、マリンコラーゲンは、コラーゲン生成促進効果のあるウシ由来のコラーゲンと比べ、単独の場合でも、ヘチマとアマチャヅルの雑種細胞抽出物を併用した場合でもヒト皮膚線維芽細胞のコラーゲン生成をより促進した。また、マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物を併用した場合、平均分子量30万の高分子マリンコラーゲンに比べ、平均分子量200〜1,500の低分子マリンコラーゲンは、ヒト皮膚線維芽細胞のコラーゲン生成をより促進した。
【0027】
実験例2 弾力改善試験1
表1で示した実施例の美容液1と比較例1〜2の美容液を用いて、弾力改善試験を実施した。試験は、健康な女性30人(37〜44才)を対象に3ヶ月行った。被験者を10人ずつ3群に分け、I群は美容液1、II群は比較例1、III群は比較例2を使用することとした。使用前に測定した肌の弾力をコントロールとし、使用後の肌の弾力と比較した。肌の弾力は、キュートメーター(Courage+Khazaka社製)を用いて測定した。
【0028】
肌の弾力の測定結果を表4に示した。この表から美容液1は優れた弾力改善効果を有していることが認められた。
【0029】
【表4】
【0030】
表4に示すように、マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物を併用することによってマリンコラーゲン、ヘチマとアマチャヅルの雑種細胞抽出物単独で配合したときより効果が高くなることが認められた。
【0031】
実験例3 弾力改善試験2
表2に示した実施例の美容液1〜5の美容液を用いて、弾力改善試験を実施した。試験は、健康な女性50人(34〜42才)を対象に3ヶ月行った。被験者を10人ずつ5群にわけ、I群は美容液1、II群は美容液2、III群は美容液3、IV群は美容液4、V群は美容液5を使用することとした。試験は実験例2 弾力改善試験1と同様の方法で実施した。
【0032】
【表5】
【0033】
表5に示すように、マリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物をあわせた配合量が乾固物に換算して本発明化粧料の全量中、0.00001重量%以上になるとき、より効果が高くなることが認められた。
【0034】
実験例4 使用試験
表1に示した実施例の美容液1と比較例1〜2の美容液を用いて、しわ、たるみ、肌の弾力について、アンケート調査を行って老化防止効果を評価した。試験は、実験例2と同様、健康な女性60人(35〜45才)を対象に3ヶ月行った。被験者を20人ずつ3群に分け、I群は美容液1、II群は比較例1、III群は比較例2を使用することとした。アンケートの評価基準は、使用前に比べて有効なものを「優」、やや有効なものを「良」、わずかに有効なものを「可」、無効なものを「不可」として評価した。アンケートの結果を表6に示した。この表から実施例のマリンコラーゲンと、ヘチマとアマチャヅルの雑種細胞抽出物を併用する美容液1は優れた老化防止効果を有していることが認められた。
【0035】
【表6】
[0001]
BACKGROUND OF THE INVENTION
TECHNICAL FIELD The present invention relates to a collagen production promoter and anti-aging cosmetic comprising a blend of marine collagen and a hybrid cell extract of loofah and hamchael.
[0002]
[Prior art]
One of the skin aging phenomena is the generation of wrinkles and a decrease in elasticity, and it is known that the decrease in collagen, which is a constituent component of the dermis, has a great influence on this. Therefore, many cosmetics blended with collagen and cosmetics blended with collagen production promoting substances have been developed.
[0003]
[Problems to be solved by the invention]
However, although conventional cosmetics containing collagen have a temporary moisturizing effect, they have not fundamentally improved the amount of collagen in the dermis. Moreover, the effect of the cosmetics which mix | blended the collagen production | generation promoting substance was not enough, and the more effective thing was desired. Accordingly, the present inventors have found that by combining a collagen production promoter in a cosmetic, the decrease in the amount of dermal collagen can be more effectively suppressed, and the generation of wrinkles and the decrease in elasticity can be suppressed. It is.
[0004]
[Means for Solving the Problems]
The present invention is a cosmetic characterized in that it contains marine collagen and a hybrid cell extract of loofah and hamchael. The marine collagen referred to in the present invention is a collagen obtained from the skin, floating bag, etc. of fish (for example, salmon, gold-eye salmon, flat eyes, salmon, salmon, salmon, etc.). That is, the marine collagen described in the present specification indicates fish-derived collagen. Recently, marine collagen has been used as a moisturizing agent, but marine collagen has hardly been studied compared to bovine-derived collagen that has been widely used in the past. This time, it has been found that marine collagen has an effect of promoting collagen production. Furthermore, it discovered that it was highly effective by the low molecular weight marine collagen by hydrolysis. The present invention also includes modified marine collagen such as succinylated collagen and phthalated collagen.
[0005]
In addition, it is known that hybrid cell extracts of loofah and amachazul inhibit collagen degradation by preventing serine protease and prevent skin aging (JP-A-11-124325). There is no known at all.
[0006]
As a result of discovering that marine collagen has an effect of promoting collagen production and researching to further enhance the effect, it has been shown that marine collagen is used together with a hybrid cell extract of marine collagen, loofah and amachazuru, and exhibits particularly remarkable effects. I found it. Furthermore, it discovered that a higher effect was exhibited by using low molecular weight marine collagen with an average molecular weight of 200 to 1,500 in combination with a hybrid cell extract of loofah and hamchael.
[0007]
Extraction and purification of marine collagen used in the present invention can be performed using a generally known method. Specifically, for example, the fish skin is crushed and then washed with water, extracted with an acid or alkaline solution, extracted with an enzyme such as pepsin, trypsin, or hyalonidase, and subjected to a known purification means such as salting out or dialysis to obtain an average molecular weight of 30. About 10,000 collagen solution can be obtained. This collagen solution may be used as it is, or may be used after treatment such as concentration, dilution and filtration, and decolorization and deodorization with activated carbon or the like, if necessary.
Furthermore, a collagen peptide having a molecular weight of about 200 to 1,500 can be obtained by adding collagenase to this collagen solution by a generally known method.
Regardless of the average molecular weight, marine collagen such as succinylated collagen or phthalated collagen obtained by chemically reacting the resulting collagen solution with succinic acid or phthalic acid can also be used.
[0008]
The marine collagen thus obtained may be used as a dried product after processing such as concentration to dryness, spray drying, freeze drying and the like.
[0009]
Said marine collagen may be used independently and may be used as a 2 or more types of mixture.
[0010]
Hybrid cell callus produced by the method of Japanese Patent Application Laid-Open No. 01-44297 can be used as the hybrid cell extract of loofah and amachazul.
[0011]
For calli of hybrid cells of loofah and hamchael, hybrid cell callus (F3, F4, F5, F6) produced by the method of JP-A-01-44297 may be used singly or as a mixture of two or more. .
[0012]
As a solvent for extracting hybrid cells of loofah and amachazul, in addition to water, for example, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols ( Glycerin, propylene glycol, 1,3-butylene glycol, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ether (Ethyl ether, propyl ether, tetrahydrofuran, etc.). Preferably, water, lower alcohol and liquid polyhydric alcohol are preferable, and water, ethanol, propylene glycol and 1,3-butylene glycol are particularly preferable. These solvents may be used alone or in combination of two or more.
[0013]
The blending amount of marine collagen used in the present invention and the hybrid cell extract of loofah and hamchael is preferably 0.00001 to 10% by weight in the total amount of the cosmetic of the present invention in terms of dry solid. If it is less than 0.00001% by weight, a sufficient effect cannot be expected. When it exceeds 10% by weight, the effect is not enhanced and it is uneconomical. The addition method may be added in advance or during the production, and may be appropriately selected in consideration of workability.
[0014]
The cosmetics of the present invention include fats and oils, waxes, hydrocarbons, fatty acids, which are components used in ordinary cosmetics, within the range that does not impair the effects of marine collagen and the hybrid cell extract of loofah and hamchael. Ingredients such as alcohol, alcohols, esters, surfactants, metal soaps, pH adjusters, preservatives, fragrances, moisturizers, powders, UV absorbers, thickeners, dyes, antioxidants, chelating agents, etc. Can be blended.
[0015]
Examples of the anti-aging cosmetic of the present invention include skin lotion, milky lotion, cream, cosmetic liquid, pack, gelling agent, cleaning agent, bath preparation, foundation, lipstick, and eye shadow.
[0016]
【Example】
Hereinafter, the present invention will be described in detail with reference to examples. However, the technical scope of the present invention is not limited by these production examples. The part of the amount shown in the examples indicates part by weight and% indicates% by weight.
[0017]
Manufacture example 1: 1 kg of marine collagen cocoon skin derived from cocoon skin was used to extract a collagen solution obtained by a conventionally known method by concentration, dilution, filtration, decolorization and deodorization treatment using activated carbon, etc. The solution was freeze-dried to obtain 140 g of collagen powder having an average molecular weight of 300,000.
Further, hot water was added to the collagen powder to denature it into gelatin, and the collagen peptide solution hydrolyzed by adding collagenase by a generally known method was freeze-dried to obtain 135 g of low molecular weight collagen peptide powder.
[0018]
Production Example 2: Hetchima-Amateur hybrid cell 50% ethanol extract 1,000 ml of 50% ethanol was added to 100 g of Hettima-Amateur hybrid cell (F5), extracted at room temperature for 7 days, filtered, and the filtrate was filtered. The mixture was concentrated and dried to obtain 7 g of 50% ethanol extract of hybrid cells of loofah and amachazuru.
[0019]
Example The cosmetic liquid 1-5 shown in Tables 1-2 and the comparative examples 1-2 can be made by changing the compounding quantity of the hybrid liquid cell extract of cosmetic liquid marine collagen and a loofah and an amachar.
[0020]
[Table 1]
[0021]
[Table 2]
[0022]
[Manufacturing method] In Tables 1 and 2, components 1 to 7 and 12 and components 8 to 11 are uniformly dissolved, and both are mixed and adjusted.
[0023]
【The invention's effect】
Next, experimental examples will be given to explain the effects of the present invention in detail.
[0024]
Experimental Example 1 Human skin fibroblasts (derived from a 6-year-old boy, passage number 6) were used as test cells, and Eagle's MEM was used as a medium in the collagen production promoting action test. Measurement of collagen production amount was carried out the uptake of 3 H- proline into the collagen protein as an indicator. A 96-well plate (Falcon 3072) was seeded with 25,000 cells / well of human dermal fibroblasts, and cultured in Eagle's MEM containing 10% FCS for 4 days under 37 ° C. and 5% CO 2 conditions. During this time, the cells became confluent. Next, marine collagen of the present invention (Production Example 1; average molecular weight 100, average molecular weight 200, average molecular weight 500, average molecular weight 1,500, average molecular weight 3,000, average molecular weight 300,000), hybrid cell extraction of loofah and amachazuru Product (Production Example 2), marine collagen (Production Example 1; average molecular weight 100, average molecular weight 200, average molecular weight 500, average molecular weight 1,500, average molecular weight 3,000, average molecular weight 300,000) + hybrid cells of loofah and amacha Serum-free Eagle's MEM (50 μg / ml L-ascorbic acid, 50 μg / ml β-aminopropionitrile, 18.5 KBq 3) prepared by adjusting the extract (Production Example 2) to a final concentration of 5 μg / ml. H-proline-containing) and cultured for 24 hours. Thereafter, a pepsin solution was immediately added to each well and incubated, and then 3 H-proline-labeled collagen was extracted. After purification by salting out, the radioactivity in the collagen protein was measured with a liquid scintillation counter (Beckman LS 5801). For each sample, 5 wells were measured, and the production amount was shown as an average value ± standard deviation. A sample-free sample was used as a control.
[0025]
[Table 3]
[0026]
As shown in Table 3, the hybrid cell extract of marine collagen + loofah and amacharu of the present invention significantly promoted collagen production of human skin fibroblasts. The effect was improved by using marine collagen and a hybrid cell extract of Hetchima and Achachar together with a mere marine collagen, a hybrid cell extract of Loofah and Achachar alone. In addition, marine collagen further promoted collagen production in human dermal fibroblasts, when used alone or in combination with a hybrid cell extract of loofah and hamchael, compared with bovine-derived collagen, which has an effect of promoting collagen production. Further, when marine collagen and a hybrid cell extract of loofah and hamchael are used in combination, low molecular weight marine collagen having an average molecular weight of 200 to 1,500 is higher than human marine collagen having an average molecular weight of 300,000. Promoted more collagen production.
[0027]
Experiment 2 Elasticity improvement test 1
The elasticity improvement test was implemented using the cosmetic liquid 1 of the Example shown in Table 1, and the cosmetic liquid of Comparative Examples 1-2. The test was conducted on 30 healthy women (37-44 years old) for 3 months. The test subjects were divided into 3 groups of 10 people, the serum I was used as the group I, the comparative example 1 was used as the group II, and the comparative example 2 was used as the group III. The skin elasticity measured before use was used as a control and compared with the skin elasticity after use. The elasticity of the skin was measured using a cute meter (Courage + Khazaka).
[0028]
The measurement results of skin elasticity are shown in Table 4. From this table, it was confirmed that the cosmetic liquid 1 has an excellent elasticity improving effect.
[0029]
[Table 4]
[0030]
As shown in Table 4, it was found that the combined use of marine collagen and a hybrid cell extract of loofah and hamchael was more effective than that of marine collagen, a hybrid cell extract of loofah and hamachal alone. .
[0031]
Experiment 3 Elasticity improvement test 2
Using the essences of essences 1 to 5 of Examples shown in Table 2, the elasticity improvement test was performed. The test was conducted for 3 months on 50 healthy women (34-42 years old). The test subjects were divided into 5 groups of 10 people, group I used essence 1, group II essence 2, group III essence 3, group IV essence 4, and group V essence 5. . The test was carried out in the same manner as in Experimental Example 2 Elasticity Improvement Test 1.
[0032]
[Table 5]
[0033]
As shown in Table 5, when the blending amount of marine collagen and the hybrid cell extract of loofah and amacharu is 0.00001% by weight or more in the total amount of the cosmetic composition of the present invention in terms of dry matter, It was observed that the effect was higher.
[0034]
Experimental Example 4 Usage Test Using the cosmetic liquid 1 of the examples shown in Table 1 and the cosmetic liquids of Comparative Examples 1 and 2, a questionnaire survey was conducted on wrinkles, sagging, and skin elasticity to evaluate the anti-aging effect. The test was conducted for 3 months on 60 healthy women (35 to 45 years old) as in Experimental Example 2. The test subjects were divided into 3 groups of 20 people, the serum I was used as the group I, the comparative example 1 was used as the group II, and the comparative example 2 was used as the group III. The evaluation criteria of the questionnaire were evaluated as “excellent” for effective items, “good” for slightly effective items, “good” for slightly effective items, and “impossible” for invalid items. The results of the questionnaire are shown in Table 6. From this table, it was confirmed that the cosmetic liquid 1 in which the marine collagen of the example and the hybrid cell extract of the loofah and the amachazul are used in combination has an excellent anti-aging effect.
[0035]
[Table 6]
Claims (4)
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