JP2000080023A - Beautifully whitening cosmetic and melanin-formation inhibitor - Google Patents
Beautifully whitening cosmetic and melanin-formation inhibitorInfo
- Publication number
- JP2000080023A JP2000080023A JP10305192A JP30519298A JP2000080023A JP 2000080023 A JP2000080023 A JP 2000080023A JP 10305192 A JP10305192 A JP 10305192A JP 30519298 A JP30519298 A JP 30519298A JP 2000080023 A JP2000080023 A JP 2000080023A
- Authority
- JP
- Japan
- Prior art keywords
- melanocytes
- production
- skin
- metallothionein
- melanin
- 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.)
- Pending
Links
- 239000002537 cosmetic Substances 0.000 title claims abstract description 13
- 239000003112 inhibitor Substances 0.000 title claims description 11
- 230000002087 whitening effect Effects 0.000 title claims description 10
- 210000002752 melanocyte Anatomy 0.000 claims abstract description 33
- 238000004519 manufacturing process Methods 0.000 claims abstract description 25
- DIGQNXIGRZPYDK-WKSCXVIASA-N (2R)-6-amino-2-[[2-[[(2S)-2-[[2-[[(2R)-2-[[(2S)-2-[[(2R,3S)-2-[[2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2R)-2-[[(2S,3S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2R)-2-[[2-[[2-[[2-[(2-amino-1-hydroxyethylidene)amino]-3-carboxy-1-hydroxypropylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxybutylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1,5-dihydroxy-5-iminopentylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxybutylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxyethylidene]amino]hexanoic acid Chemical compound C[C@@H]([C@@H](C(=N[C@@H](CS)C(=N[C@@H](C)C(=N[C@@H](CO)C(=NCC(=N[C@@H](CCC(=N)O)C(=NC(CS)C(=N[C@H]([C@H](C)O)C(=N[C@H](CS)C(=N[C@H](CO)C(=NCC(=N[C@H](CS)C(=NCC(=N[C@H](CCCCN)C(=O)O)O)O)O)O)O)O)O)O)O)O)O)O)O)N=C([C@H](CS)N=C([C@H](CO)N=C([C@H](CO)N=C([C@H](C)N=C(CN=C([C@H](CO)N=C([C@H](CS)N=C(CN=C(C(CS)N=C(C(CC(=O)O)N=C(CN)O)O)O)O)O)O)O)O)O)O)O)O DIGQNXIGRZPYDK-WKSCXVIASA-N 0.000 claims description 27
- 102000003792 Metallothionein Human genes 0.000 claims description 27
- 108090000157 Metallothionein Proteins 0.000 claims description 27
- 239000000126 substance Substances 0.000 claims description 14
- 230000008099 melanin synthesis Effects 0.000 claims description 13
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 abstract description 34
- 230000000694 effects Effects 0.000 abstract description 21
- 102000003425 Tyrosinase Human genes 0.000 abstract description 19
- 108060008724 Tyrosinase Proteins 0.000 abstract description 19
- 210000003491 skin Anatomy 0.000 abstract description 11
- 230000001939 inductive effect Effects 0.000 abstract description 5
- 239000006210 lotion Substances 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 5
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 239000006071 cream Substances 0.000 abstract description 2
- 239000002674 ointment Substances 0.000 abstract description 2
- 230000019612 pigmentation Effects 0.000 abstract description 2
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 abstract 2
- 239000000463 material Substances 0.000 abstract 2
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 description 28
- 235000005074 zinc chloride Nutrition 0.000 description 14
- 239000011592 zinc chloride Substances 0.000 description 14
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 8
- 102000002045 Endothelin Human genes 0.000 description 7
- 108050009340 Endothelin Proteins 0.000 description 7
- ZUBDGKVDJUIMQQ-UBFCDGJISA-N endothelin-1 Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)NC(=O)[C@H]1NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@@H](CC=2C=CC(O)=CC=2)NC(=O)[C@H](C(C)C)NC(=O)[C@H]2CSSC[C@@H](C(N[C@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N2)=O)NC(=O)[C@@H](CO)NC(=O)[C@H](N)CSSC1)C1=CNC=N1 ZUBDGKVDJUIMQQ-UBFCDGJISA-N 0.000 description 7
- 102400000740 Melanocyte-stimulating hormone alpha Human genes 0.000 description 6
- 101710200814 Melanotropin alpha Proteins 0.000 description 6
- 230000006698 induction Effects 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- WHNFPRLDDSXQCL-UAZQEYIDSA-N α-msh Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(N)=O)NC(=O)[C@H](CO)NC(C)=O)C1=CC=C(O)C=C1 WHNFPRLDDSXQCL-UAZQEYIDSA-N 0.000 description 6
- 229910002092 carbon dioxide Inorganic materials 0.000 description 5
- 239000001569 carbon dioxide Substances 0.000 description 5
- 239000012228 culture supernatant Substances 0.000 description 5
- 210000002510 keratinocyte Anatomy 0.000 description 5
- 238000000034 method Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 210000004748 cultured cell Anatomy 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000002516 radical scavenger Substances 0.000 description 3
- 239000002884 skin cream Substances 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 description 2
- 238000000636 Northern blotting Methods 0.000 description 2
- 208000012641 Pigmentation disease Diseases 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000000411 inducer Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- ZOOGRGPOEVQQDX-UUOKFMHZSA-N 3',5'-cyclic GMP Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=C(NC2=O)N)=C2N=C1 ZOOGRGPOEVQQDX-UUOKFMHZSA-N 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 108010078321 Guanylate Cyclase Proteins 0.000 description 1
- 102000014469 Guanylate cyclase Human genes 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 1
- 229930064664 L-arginine Natural products 0.000 description 1
- 235000014852 L-arginine Nutrition 0.000 description 1
- 239000000637 Melanocyte-Stimulating Hormone Substances 0.000 description 1
- 108010007013 Melanocyte-Stimulating Hormones Proteins 0.000 description 1
- 102000008299 Nitric Oxide Synthase Human genes 0.000 description 1
- 108010021487 Nitric Oxide Synthase Proteins 0.000 description 1
- 229940088382 Nitric oxide scavenger Drugs 0.000 description 1
- ZIIQCSMRQKCOCT-YFKPBYRVSA-N S-nitroso-N-acetyl-D-penicillamine Chemical compound CC(=O)N[C@@H](C(O)=O)C(C)(C)SN=O ZIIQCSMRQKCOCT-YFKPBYRVSA-N 0.000 description 1
- YZCKVEUIGOORGS-NJFSPNSNSA-N Tritium Chemical compound [3H] YZCKVEUIGOORGS-NJFSPNSNSA-N 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 229960002173 citrulline Drugs 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- MHUWZNTUIIFHAS-CLFAGFIQSA-N dioleoyl phosphatidic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(COP(O)(O)=O)OC(=O)CCCCCCC\C=C/CCCCCCCC MHUWZNTUIIFHAS-CLFAGFIQSA-N 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 229960004502 levodopa Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000002464 muscle smooth vascular Anatomy 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- -1 sodium nitroferricyanide Chemical compound 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229910052722 tritium Inorganic materials 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
- 235000005282 vitamin D3 Nutrition 0.000 description 1
- 239000011647 vitamin D3 Substances 0.000 description 1
- 229940021056 vitamin d3 Drugs 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Landscapes
- Cosmetics (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、メラノサイトのメ
タロチオネイン産生を誘導する物質を配合することによ
り、メラノサイトのチロシナーゼ活性を抑制することを
特徴とする美白化粧料及びメラニン生成抑制剤に関す
る。The present invention relates to a whitening cosmetic and a melanin production inhibitor characterized by suppressing the tyrosinase activity of melanocytes by adding a substance that induces the production of metallothionein by melanocytes.
【０００２】[0002]
【従来の技術】生体内ではＬ−アルギニンから一酸化窒
素合成酵素により、一酸化窒素とＬ−シトルリンが合成
される。この合成された一酸化窒素はケミカルメディエ
ーターとして働き、細胞内のグアニレートシクラーゼを
活性化してｃＧＭＰを増加させ、血管平滑筋を弛緩させ
る作用をもつことが良く知られている。一方、皮膚にお
いても一酸化窒素の刺激によりメラノサイトのチロシナ
ーゼ活性及びメラニン合成が上昇することが知られてい
る(Romero-Graillet C. et al., Journal of Bio
logical Chemistry. vol.271, NO.45, p.28052-6,
1996)。また、ＵＶ照射することによりケラチノサイ
トの一酸化窒素発生量は上昇する。このＵＶ刺激したケ
ラチノサイトとメラノサイトをコカルチャーすることに
より、メラノサイトのチロシナーゼ活性が上昇する。こ
の上昇は一酸化窒素のスカベンジャーにより抑制される
ことが知られている(Romero-Graillet C. et al.,
J. Clin. Invest. vol.99, p.635-642, 1997) 。2. Description of the Related Art In vivo, nitric oxide and L-citrulline are synthesized from L-arginine by nitric oxide synthase. It is well known that the synthesized nitric oxide acts as a chemical mediator, activates intracellular guanylate cyclase, increases cGMP, and relaxes vascular smooth muscle. On the other hand, it is known that stimulating nitric oxide also increases tyrosinase activity and melanin synthesis of melanocytes in skin (Romero-Graillet C. et al., Journal of Bio
logical Chemistry. vol.271, NO.45, p.28052-6,
1996). In addition, the amount of nitric oxide generated in keratinocytes is increased by UV irradiation. By co-culturing the UV-stimulated keratinocytes and melanocytes, the tyrosinase activity of the melanocytes increases. This increase is known to be suppressed by nitric oxide scavengers (Romero-Graillet C. et al.,
J. Clin. Invest. Vol. 99, p. 635-642, 1997).
【０００３】また、皮膚が紫外線に暴露されたとき、細
胞内では様々なサイトカイン、ケモカイン等の物質が放
出される。それら物質の中でケラチノサイトから分泌さ
れる因子でエンドセリン及びメラノサイト刺激ホルモン
（α−ＭＳＨ）がメラノサイトを活性化することが報告
されている（Imokawa G. et al., J.Invent. Derm
atol. vol.105, p.32-37,1995、Schauer E. et a
l., J.Clin.Invest.vol.93, p.2258-2262, 1994)。When the skin is exposed to ultraviolet rays, various substances such as cytokines and chemokines are released inside the cells. Among these substances, endothelin and melanocyte stimulating hormone (α-MSH), a factor secreted from keratinocytes, have been reported to activate melanocytes (Imokawa G. et al., J. Invent. Derm.
atol.vol.105, p.32-37,1995, Schauer E. et a
l., J. Clin. Invest. vol. 93, p. 2258-2262, 1994).
【０００４】チロシナーゼ活性が上昇した場合、当然の
ことながらメラニン合成が高まり、その結果、皮膚の色
素沈着、しみ、くすみといった症状が現れる。そのため
一酸化窒素等の活性化物質の刺激を軽減し、チロシナー
ゼ活性上昇を防ぐ方法が望まれていた。一般に、一酸化
窒素等によるチロシナーゼ活性上昇を防ぐにはラジカル
スカベンジャー等のチロシナーゼ活性阻害剤の使用（細
胞外からの投与）が考えられる。[0004] When tyrosinase activity increases, melanin synthesis naturally increases, and as a result, symptoms such as skin pigmentation, spots and dullness appear. Therefore, there has been a demand for a method of reducing stimulation of an activating substance such as nitric oxide and preventing an increase in tyrosinase activity. In general, use of a tyrosinase activity inhibitor such as a radical scavenger (extracellular administration) can be considered to prevent an increase in tyrosinase activity due to nitric oxide or the like.
【０００５】一方、細胞内で産生される抗酸化物質であ
るメタロチオネインは炎症時に生じるフリーラジカルの
スカベンジャーの働きがあると報告されている(Hanada
K.et al., Dermatologica, vol.179(suppl.1),
p.143, 1989)。しかし、メタロチオネインの生体内で
の役割については未解明な部分が多く、またメラノサイ
トでのメタロチオネインの存在及び産生誘導に関する報
告はない。また、メラノサイトのメタロチオネイン産生
を誘導することによる、一酸化窒素等によって上昇する
チロシナーゼ活性の抑制作用やメラニン生成抑制作用は
全く知られておらず、また、その可能性について一切の
記載や報告もされていない。On the other hand, it has been reported that metallothionein, an antioxidant produced in cells, acts as a scavenger of free radicals generated during inflammation (Hanada).
K. et al., Dermatologica, vol.179 (suppl.1),
p.143, 1989). However, the role of metallothionein in vivo is largely unknown, and there are no reports on the presence and induction of production of metallothionein in melanocytes. In addition, no inhibitory effect of tyrosinase activity or melanin production inhibitory effect, which is increased by nitric oxide or the like, by inducing the production of metallothionein by melanocytes is known at all, and there is no description or report on the possibility. Not.
【０００６】[0006]
【発明が解決しようとする課題及び課題を解決するため
の手段】しかし、ラジカルスカベンジャー等のチロシナ
ーゼ活性阻害剤は不安定なものが多く、一酸化窒素等を
スカベンジしてメラニン合成の上昇を防ごうとした場
合、大量の物質を継続的に投与する必要があり、有効な
効果を得ることは難しい。係る事情に鑑み、本発明者等
は、皮膚内部でメラノサイトが本来持っている機能を高
めることにより、一酸化窒素等をスカベンジし、メラニ
ン合成上昇を抑制することを検討した。However, many tyrosinase activity inhibitors such as radical scavengers are unstable, and scavenge nitric oxide and the like to prevent an increase in melanin synthesis. In this case, it is necessary to continuously administer a large amount of the substance, and it is difficult to obtain an effective effect. In view of such circumstances, the present inventors have studied to enhance the functions inherent to melanocytes inside the skin to scavenge nitric oxide and the like and suppress the increase in melanin synthesis.
【０００７】そこで、培養細胞系での探索を鋭意検討し
た結果、メラノサイトでのメタロチオネインの存在及び
産生を見出した。そして、メラノサイト自身のメタロチ
オネイン産生を誘導することにより、一酸化窒素、エン
ドセリン及びα−ＭＳＨ等によって上昇するメラノサイ
トのチロシナーゼ活性が抑制できることを見出し、本発
明を完成するに至ったものである。[0007] Accordingly, as a result of diligent investigation into search in a cultured cell system, the presence and production of metallothionein in melanocytes were found. Then, they have found that by inducing the production of metallothionein by melanocytes themselves, it is possible to suppress the tyrosinase activity of melanocytes, which is increased by nitric oxide, endothelin, α-MSH, and the like, and completed the present invention.
【０００８】すなわち、本発明の目的は、物質の外的投
与ではなく、皮膚内部でメラノサイトが本来持っている
機能を高めることによって美白効果を有する化粧料及び
メラニン生成抑制剤を提供するにある。そして上記課題
は、生体内のメラノサイト自身のメタロチオネイン産生
を誘導する物質を配合することにより、一酸化窒素、エ
ンドセリン及びα−ＭＳＨ等によって上昇するメラノサ
イトのチロシナーゼ活性を抑制することを特徴とする美
白化粧料及びメラニン生成抑制剤によって達成される。[0008] That is, an object of the present invention is to provide a cosmetic and melanin production inhibitor having a whitening effect by enhancing the intrinsic function of melanocytes in the skin, not by external administration of a substance. The above object is to provide a whitening makeup characterized by suppressing the tyrosinase activity of melanocytes, which is increased by nitric oxide , endothelin, α-MSH, etc., by adding a substance that induces the production of metallothionein by melanocytes themselves in a living body. Attained by additives and melanin production inhibitors.
【０００９】[0009]
【発明の実施の形態】以下、本発明の構成について詳説
する。DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The configuration of the present invention will be described below in detail.
【００１０】本発明に用いられるメタロチオネイン産生
誘導剤としては、メラノサイトのメタロチオネイン産生
を誘導する物質であれば、特に限定されるものではな
い。例えば、塩化亜鉛等を挙げることができる。[0010] The metallothionein production inducer used in the present invention is not particularly limited as long as it is a substance that induces the production of metallothionein in melanocytes. For example, zinc chloride and the like can be mentioned.
【００１１】本発明の美白化粧料及びメラニン生成抑制
剤の使用形態としては、皮膚外用剤があり、例えば軟
膏、クリーム、ローション、貼付剤等が挙げられる。外
用剤の基剤としては、公知の外用基剤で良く、特に限定
されない。本発明の美白化粧料及びメラニン生成抑制剤
に対するメタロチオネイン産生誘導物質の配合量として
は、組成物全量に対して０．００００１〜１０重量％が
好ましい。０．００００１重量％未満では本発明の効果
を奏しない場合があり、１０重量％を超えて配合しても
配合量に見合った効果が得られない場合がある。The form of use of the whitening cosmetic and melanin production inhibitor of the present invention includes an external preparation for the skin, such as an ointment, a cream, a lotion, and a patch. The base of the external preparation may be a known external base and is not particularly limited. The compounding amount of the metallothionein production-inducing substance with respect to the whitening cosmetic and the melanin production inhibitor of the present invention is preferably 0.00001 to 10% by weight based on the total amount of the composition. If the amount is less than 0.00001% by weight, the effect of the present invention may not be exhibited. Even if the amount exceeds 10% by weight, the effect corresponding to the amount may not be obtained.
【００１２】[0012]
【実施例】以下、本発明の試験例を示す。 試験例１ メタロチオネイン産生誘導の確認 （１）方法The test examples of the present invention are shown below. Test Example 1 Confirmation of metallothionein production induction (1) Method
【００１３】メラノサイトをＭＣＤＢ１５３培地にて１
５０×１０4 個／ｍｌに調製し、６０ｍｍプレート
（ファルコン社製）に４ｍｌずつ播種し、９５％空気
（Ｖ／Ｖ）−５％（Ｖ／Ｖ）炭酸ガスの雰囲気下、３７
℃で１日間静置培養した。[0013] Melanocytes were cultured in MCDB153 medium for 1 hour.
It was adjusted to 50 × 10 4 cells / ml, and 4 ml of each was seeded on a 60 mm plate (manufactured by Falcon).
The culture was allowed to stand at room temperature for 1 day.
【００１４】培養上清を吸引除去し、生体内でメタロチ
オネイン産生を誘導すると報告されている塩化亜鉛（関
東化学社製）を０、５０、７０、１００μＭ添加した培
地を４ｍｌずつ各シャーレに加えた。このプレートを９
５％空気（Ｖ／Ｖ）−５％（Ｖ／Ｖ）炭酸ガスの雰囲気
下、３７℃で１日間静置培養した。また、ビタミンＤ3
（メルク社製）を１μＭ添加したものについても試験
を行った。尚、ケラチノサイトについても、ＭＣＤＢ１
５３培地にて２０×１０4 個／ｍｌに調製し、６０ｍ
ｍプレート（ファルコン社製）に４ｍｌずつ播種して、
その他は同様に行った。The culture supernatant was removed by suction, and 4 ml of a medium containing 0, 50, 70, and 100 μM of zinc chloride (manufactured by Kanto Kagaku), which is reported to induce metallothionein production in vivo, was added to each dish. . 9 this plate
The cells were cultured at 37 ° C. for 1 day in an atmosphere of 5% air (V / V) -5% (V / V) carbon dioxide. Also, vitamin D3
(Merck) was also tested. For keratinocytes, MCDB1
Prepared at 20 × 10 4 cells / ml with 53 medium,
Seed 4 ml each on an m plate (Falcon),
Others were performed similarly.
【００１５】培養細胞からＴｏｔａｌ ＲＮＡを抽出
し、ノザンブロット解析を行った。 （２）結果[0015] Total RNA was extracted from the cultured cells and subjected to Northern blot analysis. (2) Result
【００１６】結果を図１及び表１に示す。The results are shown in FIG. 1 and Table 1.
【００１７】[0017]
【表１】 [Table 1]
【００１８】±：ほとんど検出されない。 ++：強いシグナルが検出された。±: Almost no detection. ++: A strong signal was detected.
【００１９】図１より明らかなように塩化亜鉛を加える
ことによりメラノサイトにおいてもメタロチオネインの
ｍＲＮＡ合成量の上昇が認められ、塩化亜鉛がメタロチ
オネイン産生を誘導することが確認できた。As is apparent from FIG. 1, the addition of zinc chloride also increased the amount of metallothionein mRNA synthesis in melanocytes, confirming that zinc chloride induces metallothionein production.
【００２０】また、表１のようにケラチノサイトでメタ
ロチオネイン産生の誘導が認められたビタミンＤ3 は
メラノサイトではメタロチオネイン産生の誘導が認めら
れず、細胞の種類によりメタロチオネイン産生の誘導形
態は異なっていた。つまり、このことより、すべてのメ
タロチオネイン産生誘導物質がメラノサイトにおいて有
効であるとは言えないことが判る。Further, vitamin D 3 induction of metallothionein production in keratinocytes was observed as shown in Table 1 was not observed the induction of metallothionein production in melanocytes, derived forms metallothionein production by cell type was different. That is, this indicates that not all metallothionein production inducers are effective in melanocytes.
【００２１】試験例２ メタロチオネイン産生の誘導
時間 （１）方法 メラノサイトをＭＣＤＢ１５３培地にて１５０×１０4
個／ｍｌに調製し、６０ｍｍプレート（ファルコン社
製）に４ｍｌずつ播種し、９５％空気（Ｖ／Ｖ）−５％
（Ｖ／Ｖ）炭酸ガスの雰囲気下、３７℃で１日間静置培
養した。Test Example 2 Induction time of metallothionein production (1) Method Melanocytes were cultured in MCDB153 medium at 150 × 10 4.
Per ml, seeded in 4 ml each on a 60 mm plate (Falcon), and 95% air (V / V) -5%
(V / V) The cells were cultured at 37 ° C. for 1 day in an atmosphere of carbon dioxide.
【００２２】培養上清を吸引除去し、塩化亜鉛１００μ
Ｍ添加した培地を４ｍｌずつ各シャーレに加えた。この
プレートを９５％空気（Ｖ／Ｖ）−５％（Ｖ／Ｖ）炭酸
ガスの雰囲気下、３７℃で静置培養した。The culture supernatant is removed by suction, and zinc chloride (100 μl) is removed.
4 ml of the medium supplemented with M was added to each dish. The plate was incubated at 37 ° C. in an atmosphere of 95% air (V / V) -5% (V / V) carbon dioxide.
【００２３】０、６、１２、２４、４８、７２時間培養
後、培養細胞から蛋白を抽出し、ウエスタンブロット解
析を行った。 （２）結果After culturing for 0, 6, 12, 24, 48 and 72 hours, proteins were extracted from the cultured cells and subjected to Western blot analysis. (2) Result
【００２４】結果を表２に示す。The results are shown in Table 2.
【００２５】[0025]
【表２】 [Table 2]
【００２６】−：検出されず。 ＋：シグナルが検出された。-: Not detected. +: A signal was detected.
【００２７】表２より明らかなように塩化亜鉛を加えて
２４時間後にメタロチオネインの産生が誘導されている
ことがわかった。As apparent from Table 2, it was found that the production of metallothionein was induced 24 hours after the addition of zinc chloride.
【００２８】試験例３ チロシナーゼ活性測定 （１）方法 メラノサイトをＭＣＤＢ１５３培地にて６０×１０4
個／ｍｌに調製し、２４穴プレート（ファルコン社製）
に１ｍｌずつ播種し、９５％空気（Ｖ／Ｖ）−５％（Ｖ
／Ｖ）炭酸ガスの雰囲気下、３７℃で１日間静置培養し
た。Test Example 3 Measurement of Tyrosinase Activity (1) Method Melanocytes were cultured in MCDB153 medium at 60 × 10 4.
24ml plate (Falcon)
1% each, and 95% air (V / V) -5% (V
/ V) The cells were cultured at 37 ° C. for 1 day in a carbon dioxide atmosphere.
【００２９】培養上清を吸引除去し、塩化亜鉛を０、５
０、７０、１００μＭ添加した培地を１ｍｌずつ各ウェ
ルに加えた。尚、コントロールとして精製水を添加し
た。このプレートを９５％空気（Ｖ／Ｖ）−５％（Ｖ／
Ｖ）炭酸ガスの雰囲気下、３７℃で１日間静置培養し、
メタロチオネイン産生を誘導した。The culture supernatant is removed by suction, and zinc chloride
1 ml of the medium supplemented with 0, 70 and 100 μM was added to each well. In addition, purified water was added as a control. The plate is subjected to 95% air (V / V) -5% (V / V).
V) Static culture at 37 ° C. for 1 day in an atmosphere of carbon dioxide,
Metallothionein production was induced.
【００３０】２４時間誘導後、一酸化窒素発生剤（ＳＮ
ＡＰ：S-nitroso-N-acetylpenicillamine ）を２００
μＭ、エンドセリン１０ｎＭ、又はα−ＭＳＨを終濃度
５００ｎｇ／ｍｌ添加して、培養を４日間行った。尚、
コントロールとしてジメチルスルホキシドを添加した。
また、培養３日目に１μＣｉの〔 3Ｈ〕−チロシン
（第一化学薬品社製）を培地に添加した。また、一酸化
窒素発生剤としてはsodium nitroferricyanide等を用
いることもできる。After induction for 24 hours, nitric oxide generator (SN)
AP: S-nitroso-N-acetylpenicillamine) 200
μM, 10 nM of endothelin, or α-MSH was added at a final concentration of 500 ng / ml, and culturing was performed for 4 days. still,
Dimethyl sulfoxide was added as a control.
On the third day of culture, 1 μCi of [ 3 H] -tyrosine (Daiichi Pure Chemicals) was added to the medium. Further, sodium nitroferricyanide or the like can also be used as a nitric oxide generator.
【００３１】培養終了後、〔 3Ｈ〕−チロシンがチロ
シナーゼによりＤＯＰＡ［３−（３，４−Ｄｉｈｙｄｒ
ｏｘｙｐｈｅｎｙｌ）−α−ａｌａｎｉｎｅ］に変換さ
れる際に培養上清中に出てくるトリチウム水の放射活性
を、液体シンチレーションカウンターにて測定した。
尚、培養上清中に未反応で残っている〔 3Ｈ〕−チロ
シンは活性炭に吸着させ除去した。After completion of the culture, [ 3 H] -tyrosine was converted to DOPA [3- (3,4-Dihydrr) by tyrosinase.
[oxyphenyl) -α-alanine], and the radioactivity of tritium water which appeared in the culture supernatant when converted to a liquid scintillation counter was measured.
[ 3 H] -tyrosine remaining unreacted in the culture supernatant was removed by adsorption to activated carbon.
【００３２】（２）結果 結果を図２、３及び４に示す。図２、３及び４より明ら
かなように一酸化窒素発生剤（ＳＮＡＰ）、エンドセリ
ン及びα−ＭＳＨ添加前に塩化亜鉛を加えてメタロチオ
ネイン産生を誘導しておくと上昇したチロシナーゼ活性
の抑制効果が認められた。(2) Results The results are shown in FIGS. As is clear from FIGS. 2, 3 and 4, addition of zinc chloride prior to the addition of nitric oxide generator (SNAP), endothelin and α-MSH to induce metallothionein production showed an increased inhibitory effect on tyrosinase activity. Was done.
【００３３】以下、本発明の美白化粧料及びメラニン生
成抑制剤の応用例を示す。Hereinafter, application examples of the whitening cosmetic and the melanin production inhibitor of the present invention will be described.
【００３４】応用例１（スキンクリーム） 塩化亜鉛を表３の組成で配合し美白化粧料（スキンクリ
ーム）を調製した。Application Example 1 (Skin cream) Zinc chloride was blended with the composition shown in Table 3 to prepare a whitening cosmetic (skin cream).
【００３５】[0035]
【表３】 [Table 3]
【００３６】（２）調製法 Ａ成分及びＢ成分を各々８０℃に加熱溶解した後、混合
して、攪拌しつつ３０℃まで冷却して、スキンクリーム
を調製した。(2) Preparation method The components A and B were each heated and dissolved at 80 ° C, mixed, and cooled to 30 ° C with stirring to prepare a skin cream.
【００３７】応用例２（ローション） 塩化亜鉛を表４の組成で配合し、美白化粧料（ローショ
ン）を調製した。Application Example 2 (Lotion) Zinc chloride was blended with the composition shown in Table 4 to prepare a whitening cosmetic (lotion).
【００３８】[0038]
【表４】 [Table 4]
【００３９】（２）調製法 Ａ成分及びＢ成分をそれぞれ混合溶解し、ＢをＡに加え
て混合攪拌してローションを調製した。(2) Preparation Method A component and B component were mixed and dissolved, B was added to A, and the mixture was stirred to prepare a lotion.
【００４０】[0040]
【発明の効果】以上の如く、本発明により、皮膚内部で
メラノサイト自身のメタロチオネイン産生を誘導させる
物質によって、一酸化窒素、エンドセリン又はα−ＭＳ
Ｈ等によって上昇するメラノサイトのチロシナーゼ活性
を抑制することができ、メラニン生成抑制剤として有用
なことは明らかである。そして、皮膚の色素沈着、し
み、くすみの改善ができることは明らかである。As described above, according to the present invention, nitric oxide, endothelin or α-MS can be produced by using a substance that induces melanocyte production of metallothionein in the skin.
It is possible to suppress the tyrosinase activity of melanocytes, which is increased by H or the like, and it is clear that it is useful as a melanin production inhibitor. It is clear that pigmentation, spots and dullness of the skin can be improved.
【図１】塩化亜鉛による、メラノサイトのメタロチオネ
インｍＲＮＡ合成量の上昇を示したノザンブロット解析
を示した図である。FIG. 1 is a diagram showing Northern blot analysis showing an increase in the amount of metallothionein mRNA synthesis in melanocytes by zinc chloride.
【図２】一酸化窒素によって上昇したメラノサイトのチ
ロシナーゼ活性の、塩化亜鉛による抑制効果を示した図
である。FIG. 2 is a graph showing the inhibitory effect of zinc chloride on tyrosinase activity of melanocytes increased by nitric oxide.
【図３】エンドセリンによって上昇したメラノサイトの
チロシナーゼ活性の、塩化亜鉛による抑制効果を示した
図である。FIG. 3 shows the inhibitory effect of zinc chloride on the tyrosinase activity of melanocytes increased by endothelin.
【図４】α−ＭＳＨによって上昇したメラノサイトのチ
ロシナーゼ活性の、塩化亜鉛による抑制効果を示した図
である。FIG. 4 is a graph showing the inhibitory effect of zinc chloride on the tyrosinase activity of melanocytes increased by α-MSH.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 五十嵐 滋 神奈川県小田原市寿町５丁目３番28号 鐘 紡株式会社基礎科学研究所内 (72)発明者 堀越 俊雄 神奈川県小田原市寿町５丁目３番28号 鐘 紡株式会社基礎科学研究所内 Ｆターム(参考） 4C083 AA082 AB332 AC072 AC102 AC122 AC242 AC442 AC472 AC482 AC782 AC792 AD352 AD512 BB51 CC01 CC04 CC05 DD23 DD31 EE16 FF05 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Shigeru Igarashi 5-3-28 Kotobuki-cho, Odawara-shi, Kanagawa Prefecture Kanebo Co., Ltd. Basic Research Institute (72) Inventor Toshio Horikoshi 5-3-1 Kotobuki-cho, Odawara-shi, Kanagawa No. 28 Kanebo Co., Ltd. Basic Science Laboratory F-term (reference) 4C083 AA082 AB332 AC072 AC102 AC122 AC242 AC442 AC472 AC482 AC782 AC792 AD352 AD512 BB51 CC01 CC04 CC05 DD23 DD31 EE16 FF05
Claims (2)
誘導する物質を配合することを特徴とする美白化粧料。1. A whitening cosmetic comprising a substance that induces metallothionein production of melanocytes.
誘導する物質を配合することを特徴とするメラニン生成
抑制剤。2. A melanin production inhibitor comprising a substance that induces metallothionein production of melanocytes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10305192A JP2000080023A (en) | 1998-06-30 | 1998-10-27 | Beautifully whitening cosmetic and melanin-formation inhibitor |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18391398 | 1998-06-30 | ||
| JP10-183913 | 1998-06-30 | ||
| JP10305192A JP2000080023A (en) | 1998-06-30 | 1998-10-27 | Beautifully whitening cosmetic and melanin-formation inhibitor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2000080023A true JP2000080023A (en) | 2000-03-21 |
Family
ID=26502176
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP10305192A Pending JP2000080023A (en) | 1998-06-30 | 1998-10-27 | Beautifully whitening cosmetic and melanin-formation inhibitor |
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| Country | Link |
|---|---|
| JP (1) | JP2000080023A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003532644A (en) * | 2000-04-28 | 2003-11-05 | ロレアル | Plant extracts of Vitis vinifera species as NO synthase inhibitors and their use |
| JP2017067612A (en) * | 2015-09-30 | 2017-04-06 | 株式会社ナリス化粧品 | Method and kit for screening material |
| JP2021162359A (en) * | 2020-03-30 | 2021-10-11 | 株式会社ナリス化粧品 | Screening method for keratin plug formation preventive / ameliorating agents |
-
1998
- 1998-10-27 JP JP10305192A patent/JP2000080023A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003532644A (en) * | 2000-04-28 | 2003-11-05 | ロレアル | Plant extracts of Vitis vinifera species as NO synthase inhibitors and their use |
| JP2017067612A (en) * | 2015-09-30 | 2017-04-06 | 株式会社ナリス化粧品 | Method and kit for screening material |
| JP2021162359A (en) * | 2020-03-30 | 2021-10-11 | 株式会社ナリス化粧品 | Screening method for keratin plug formation preventive / ameliorating agents |
| JP7024004B2 (en) | 2020-03-30 | 2022-02-22 | 株式会社ナリス化粧品 | Screening method for preventive / ameliorating agents for keratin plug formation |
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