JP3816346B2 - SCF binding inhibitor - Google Patents
SCF binding inhibitor Download PDFInfo
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- JP3816346B2 JP3816346B2 JP2001103203A JP2001103203A JP3816346B2 JP 3816346 B2 JP3816346 B2 JP 3816346B2 JP 2001103203 A JP2001103203 A JP 2001103203A JP 2001103203 A JP2001103203 A JP 2001103203A JP 3816346 B2 JP3816346 B2 JP 3816346B2
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Description
【0001】
【発明の属する技術分野】
本発明は、細胞表面のc−kitレセプターに対するステムセルファクター(Stem cell factor、以下「SCF」という)の結合を阻害するSCF結合阻害剤に関する。
【0002】
【従来の技術及び発明が解決しようとする課題】
SCFは、造血幹細胞の表面に発現しているc−kitレセプターのリガンドであり、造血細胞の増殖、分化を促す膜結合型の増殖因子として知られている。近年、c−kitが造血細胞の他に、肥満細胞や生殖細胞等の表面にも発現していることが明らかになり、SCFが肥満細胞上のc−kitレセプターに結合することにより、肥満細胞が遊走、分化・増殖し、これにより喘息やアトピー性皮膚炎等の即時型アレルギーが引き起こされることが報告されている(J.Immunol 1996 May 15;156(10),3945-3951)。
【0003】
従って、肥満細胞表面上のc−kitレセプターとSCFとの結合を特異的に阻害することができれば、斯かる細胞の分化・増殖に起因するアレルギー疾患の予防及び治療が可能となる。
【0004】
しかしながら、細胞表面上のc−kitレセプターに対するSCFの結合を阻止する物質については、これまでに全く知られていない。
【0005】
本発明の目的は、細胞表面、特に肥満細胞表面上のc−kitレセプターに対するSCFの結合を阻害し、医薬又は化粧料として有用なSCF結合阻害剤を提供することにある。
【0006】
【課題を解決するための手段】
本発明者らは、細胞表面上のc−kitレセプターに対するSCFの結合を特異的に阻害する天然物を探索したところ、特定の植物又はその抽出物に、SCF結合阻害活性があり、当該細胞の分化・増殖に起因する疾患の予防・治療効果を有する医薬又は化粧料として有用であることを見出した。
【0007】
すなわち本発明は、ノバラ及びユリから選ばれる植物又はそれらの抽出物からなるSCF結合阻害剤を提供するものである。
【0008】
【発明の実施の形態】
本発明のSCF結合阻害剤とは、細胞表面、特に肥満細胞表面上のc−kitレセプターに対するSCFの結合を特異的に阻害し、当該細胞の分化・増殖に起因する疾患、例えば喘息、枯草熱、アトピー性皮膚炎等のアレルギー疾患に対して予防・治療効果を有するものをいう。
【0009】
本発明におけるカンゾウとは、マメ科(Leguminosae)のGlycyrrhiza uralensis Fischer, Glycyrrhiza glabra L.またはその他同属植物の根及びストロンを、アスパラサスリネアリスとは、マメ科(Leguminosae)のAspalathus linearis(N.L.Burm.)R.Dahlgrの全草を、アセンヤクとは、アカネ科(Rubiaceae)のUncaria gambir ROXB.の葉及び若枝から得た乾燥水製エキスを、ブッチャーブルームとは、ユリ科(Liliaceae)のRuscus aculeatus Linneの根茎を、シコンとは、ムラサキ科(Boraginaceae)のムラサキLithospermum erythrorhizon Siebold et Zuccariniの根を、エンメイソウとは、シソ科(Labiatae)のヒキオコシ Isodon japonicus Haraの茎葉を、トウガラシとはナス科(Solanaceae)のトウガラシ Capsicum annuum L. 又はその変種の果実を、ウコンとは、ショウガ科(Zingiberaceae)のウコン Curcuma domestica Valetonの根茎を、コンフリーとは、ムラサキ科(Boraginaceae)のSymphytum officinale Linneの葉を、ノバラとは、バラ科(Rosaceae)のRose canina Linneの果実を、ユリとは、ユリ科(Liliaceae)のユリ Lilium candidumを、チャとは、ツバキ科(Theaceae)のThea sinensis Linneの葉を、チョウジとは、フトモモ科(Myrtaceae)のEugenla caryophyllata THUNB.を、ツバキとは、ツバキ科のCamellia japonica Linneの花をそれぞれ意味する。
【0010】
本発明における上記植物は、その植物の全草、葉、樹皮、枝、果実又は根等をそのまま又は粉砕して用いることができるが、カンゾウ、シコンについては根を、アスパラサスリネアリス、エンメイソウについては全草を、アセンヤク、コンフリー、チャについては葉を、ブッチャーブルーム、ウコンについては根茎を、トウガラシ、ノバラについては果実を、ユリについては球根を、チョウジについてはつぼみを、ツバキについては花を使用することが好ましい。
また、本発明における抽出物とは、更にこれを常温又は加温下にて抽出するか又はソックスレー抽出器等の抽出器具を用いて抽出することにより得られる各種溶媒抽出液、その希釈液、その濃縮液又はその乾燥末を意味するものである。ここで抽出物は、2種以上の植物から得られたものであってもよい。
【0011】
本発明の植物抽出物を得るために用いられる抽出溶剤としては、極性溶剤、非極性溶剤のいずれをも使用することができる。例えば、水;メタノール、エタノール、プロパノール、ブタノール等のアルコール類;プロピレングリコール、ブチレングリコール等の多価アルコール類;アセトン、メチルエチルケトン等のケトン類;酢酸メチル、酢酸エチル等のエステル類;テトラヒドロフラン、ジエチルエーテル等の鎖状及び環状エーテル類;ポリエチレングリコール等のポリエーテル類;スクワラン、ヘキサン、シクロヘキサン、石油エーテル等の炭化水素類;トルエン等の芳香族炭化水素類;ジクロロメタン、クロロホルム、ジクロロエタン等のハロゲン化炭化水素類;及び二酸化炭素等が挙げられ、これらは混合物として用いることができる。
【0012】
上記の植物抽出物は、そのまま用いることもできるが、当該抽出物を希釈、濃縮若しくは凍結乾燥した後、粉末又はペースト状に調製して用いることもできる。
また、上記の植物抽出物は、クロマトグラフィー液々分配等の分離技術により、上記抽出物から不活性な夾雑物を除去して用いることもできる。
【0013】
これらの植物又はその抽出物は、後記実施例に示すように優れたSCF結合阻害活性を有する。従って、これを配合した製剤は、喘息、枯草熱、アトピー性皮膚炎等のアレルギー疾患に対して予防・治療効果を有する医薬又は化粧料として有用である。
【0014】
本発明のSCF結合阻害剤を医薬として配合する場合には、錠剤、カプセル剤等の内服剤、軟膏、水剤、エキス剤、ローション剤、乳剤等の外用剤、注射剤とすることができ、中でも外用医薬品としての使用が好ましい。
【0015】
また、化粧料として配合する場合は、種々の形態、例えば、油中水型又は水中油型の乳化化粧料、クリーム、ローション、ジェル、フォーム、エッセンス、ファンデーション、パック、スティック、パウダー等とすることができ、本発明の植物又はその抽出物の他に、化粧料成分として一般に使用されている油分、界面活性剤、紫外線吸収剤、アルコール類、キレート剤、pH調整剤、防腐剤、増粘剤、色素類、香料、各種皮膚栄養剤等を任意に組合せて配合することができる。
【0016】
医薬又は化粧料における本発明の植物又はその抽出物の配合量は、乾燥物として通常全組成の0.00001〜1重量%、特に0.0001〜0.1重量%が好ましい。
【0017】
【実施例】
以下、実施例により本発明を具体的に説明する。
製造例1 チョウジ抽出物の製造
チョウジのつぼみを乾燥し、細切した後、その150gに50%(v/v)エタノール水溶液1500mLを加え、室温で7日間抽出した後、ろ過した。このものを減圧濃縮し、さらに水7500mLを加えて減圧濃縮した後、50%(v/v)エタノール水溶液1500mLに溶かした。ここに活性炭36gを加え、室温で6時間攪拌した後、ろ過し、チョウジ抽出物を得た。収量1200mL、蒸発残分1.5%。
【0018】
製造例2
製造例1に準じて下記表1に示す各植物抽出物を調製した。
【0019】
【表1】
【0020】
実施例1 SCF結合阻害活性
24穴プレート(岩城商事)にヒト培養メラノサイト(三光純薬)を播種し、コンフルエントになるまで培養した。その後、メラノサイト増殖培地から0.05%アルブミン(和光純薬)を含むRPMI1640(Gibco社)に交換し、IODO-GEN lodination Reagent(PIERCE社)を用いて、PIERCE社の示すプロトコ―ルに従い、125Iでラベリングしたヒト組み換えSCF(IBL社)を終濃度で1nMの表2に示す各植物抽出物をそれぞれ終濃度1%で添加した。125I−SCFに対してその100倍量の未標識SCFを加えることで非特異的結合量を求めた。
90分間インキュベーション後、非結合SCFを除去するため、反応終了後、氷冷RPMI1640を用いて細胞の洗浄を行った。細胞を2M NaOHで溶解し、メラノサイト上のc−kitへの結合を常法に従い、γ−カウンターを用いて測定した。いずれも総結合量から非特異的結合量を差し引いたものをSCF/c−kitの特異的結合量とした。結果を表2に併せて示す。
【0021】
【表2】
【0022】
表2に示したとおり、本発明の植物抽出物は、c−kitに対するSCFの結合阻害活性を有することが認められた。
【0023】
【発明の効果】
本発明の植物又は植物抽出物は、細胞表面のc−kitに対するSCFの結合を阻害することから、喘息、枯草熱、アトピー性皮膚炎等のアレルギー疾患に対して予防・治療効果を有する医薬又は化粧料として使用することができる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an SCF binding inhibitor that inhibits the binding of a stem cell factor (hereinafter referred to as “SCF”) to a cell surface c-kit receptor.
[0002]
[Prior art and problems to be solved by the invention]
SCF is a ligand for the c-kit receptor expressed on the surface of hematopoietic stem cells, and is known as a membrane-bound growth factor that promotes proliferation and differentiation of hematopoietic cells. In recent years, it has become clear that c-kit is expressed not only on hematopoietic cells but also on the surface of mast cells, germ cells, etc., and by binding SCF to the c-kit receptor on mast cells, mast cells Have been reported to cause migration, differentiation / proliferation, and immediate allergies such as asthma and atopic dermatitis (J. Immunol 1996 May 15; 156 (10), 3945-3951).
[0003]
Therefore, if the binding between the c-kit receptor on the mast cell surface and SCF can be specifically inhibited, allergic diseases caused by the differentiation and proliferation of such cells can be prevented and treated.
[0004]
However, no substance has been known so far that inhibits the binding of SCF to the c-kit receptor on the cell surface.
[0005]
An object of the present invention is to provide an SCF binding inhibitor that inhibits the binding of SCF to a c-kit receptor on a cell surface, particularly a mast cell surface, and is useful as a medicine or a cosmetic.
[0006]
[Means for Solving the Problems]
When the present inventors searched for a natural product that specifically inhibits the binding of SCF to the c-kit receptor on the cell surface, a specific plant or an extract thereof has an SCF binding inhibitory activity, and It has been found that it is useful as a medicine or cosmetic having a preventive / therapeutic effect on diseases caused by differentiation and proliferation.
[0007]
That is, the present invention provides an SCF binding inhibitor comprising a plant selected from Novara and lily or an extract thereof.
[0008]
DETAILED DESCRIPTION OF THE INVENTION
The SCF binding inhibitor of the present invention specifically inhibits the binding of SCF to the c-kit receptor on the surface of cells, particularly on the surface of mast cells, and causes diseases caused by differentiation / proliferation of the cells such as asthma and hay fever. It means a substance having a preventive / therapeutic effect on allergic diseases such as atopic dermatitis.
[0009]
In the present invention, licorice refers to the roots and strons of Glycyrrhiza uralensis Fischer, Glycyrrhiza glabra L. or other related plants of legumes (Leguminosae), and Aspalathus linearis (NLBurm.) R of leguminosae. The whole plant of Dahlgr, Asenyaku is a dried water extract from leaves and young branches of Rubiaceae's Uncaria gambir ROXB. Butcher bloom is a rhizome of Ruscus aculeatus Linne of Liliaceae , Shikon refers to the roots of Lithospermum erythrorhizon Siebold et Zuccarini of the Boraginaceae, Enmeiso refers to the leaves of the Labiatae Isodon japonicus Hara, Pepper refers to the Solanaceae Capsicum annuum L. or its varieties, turmeric is the curcuma domestica Valeton rhizome from Zingiberaceae. -Is the leaf of Symphytum officinale Linne from the Boraginaceae, the rose is the fruit of Rose canina Linne from the Rosaceae, and the lily is the Lilium candidum from the Liliumceae, Refers to the leaves of Thea sinensis Linne from Theaceae, clove refers to Eugenla caryophyllata THUNB. From Myrtaceae, and camellia refers to Camellia japonica Linne flowers from Camellia.
[0010]
The above plant in the present invention can be used as it is or after pulverizing whole plants, leaves, bark, branches, fruits or roots of the plant, but for licorice, lion, root, asparagus sulinaria, emiandia Use whole grass, leaves for asenyaku, comfrey and tea, rhizome for butcher bloom, turmeric, fruit for capsicum and wild rose, bulb for lily, bud for clove and flower for camellia It is preferable to do.
In addition, the extract in the present invention is further extracted at room temperature or under heating, or various solvent extracts obtained by extraction using an extractor such as a Soxhlet extractor, diluted solutions thereof, It means a concentrated liquid or its dry powder. Here, the extract may be obtained from two or more kinds of plants.
[0011]
As the extraction solvent used for obtaining the plant extract of the present invention, either a polar solvent or a nonpolar solvent can be used. For example, water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; tetrahydrofuran and diethyl ether Linear and cyclic ethers such as polyethylene; polyethers such as polyethylene glycol; hydrocarbons such as squalane, hexane, cyclohexane and petroleum ether; aromatic hydrocarbons such as toluene; halogenated carbonization such as dichloromethane, chloroform and dichloroethane Hydrogens; and carbon dioxide, and the like, which can be used as a mixture.
[0012]
The above plant extract can be used as it is, but it can also be used after being diluted, concentrated or lyophilized, and then prepared into a powder or paste.
The plant extract can also be used after removing inactive impurities from the extract by a separation technique such as chromatographic liquid-liquid distribution.
[0013]
These plants or extracts thereof have an excellent SCF binding inhibitory activity as shown in Examples below. Accordingly, a preparation containing this is useful as a pharmaceutical or cosmetic material having a preventive / therapeutic effect on allergic diseases such as asthma, hay fever, and atopic dermatitis.
[0014]
When the SCF binding inhibitor of the present invention is formulated as a medicine, it can be used as an internal preparation such as tablets and capsules, an ointment, a liquid agent, an extract, a lotion, an emulsion, an external preparation, and an injection. Among them, use as an external medicine is preferable.
[0015]
In addition, when blended as cosmetics, various forms such as water-in-oil or oil-in-water emulsified cosmetics, creams, lotions, gels, foams, essences, foundations, packs, sticks, powders, etc. In addition to the plant of the present invention or extract thereof, oils, surfactants, ultraviolet absorbers, alcohols, chelating agents, pH adjusters, preservatives, thickeners commonly used as cosmetic ingredients , Pigments, fragrances, various skin nutrients and the like can be combined in any combination.
[0016]
The blending amount of the plant of the present invention or the extract thereof in medicine or cosmetic is usually 0.00001 to 1% by weight, particularly preferably 0.0001 to 0.1% by weight of the total composition as a dry product.
[0017]
【Example】
Hereinafter, the present invention will be described specifically by way of examples.
Production Example 1 Production of Clove Extract After the clove bud was dried and chopped, 1500 mL of a 50% (v / v) ethanol aqueous solution was added to 150 g thereof, followed by extraction at room temperature for 7 days, followed by filtration. This was concentrated under reduced pressure, further added with 7500 mL of water, concentrated under reduced pressure, and then dissolved in 1500 mL of 50% (v / v) ethanol aqueous solution. The activated carbon 36g was added here, and it stirred at room temperature for 6 hours, Then, it filtered, and obtained the clove extract. Yield 1200 mL, evaporation residue 1.5%.
[0018]
Production Example 2
Each plant extract shown in Table 1 below was prepared according to Production Example 1.
[0019]
[Table 1]
[0020]
Example 1 SCF binding inhibitory activity A 24-well plate (Iwaki Corporation) was seeded with human cultured melanocytes (Sanko Junyaku) and cultured until confluent. Then, replace the RPMI1640 (Gibco Ltd.) containing 0.05% albumin melanocytes growth media (Wako Pure Chemical), using IODO-GEN lodination Reagent (PIERCE Inc.), protocol indicated by PIERCE - According le, 125 Human recombinant SCF (IBL) labeled with I was added at a final concentration of 1 nM to each plant extract shown in Table 2 at a final concentration of 1%. The amount of non-specific binding was determined by adding 100 times the amount of unlabeled SCF to 125 I-SCF.
After incubation for 90 minutes, cells were washed with ice-cold RPMI 1640 after the reaction to remove unbound SCF. The cells were lysed with 2M NaOH, and binding to c-kit on melanocytes was measured using a γ-counter according to a conventional method. In any case, the specific binding amount of SCF / c-kit was obtained by subtracting the non-specific binding amount from the total binding amount. The results are also shown in Table 2.
[0021]
[Table 2]
[0022]
As shown in Table 2, it was confirmed that the plant extract of the present invention has SCF binding inhibitory activity against c-kit.
[0023]
【The invention's effect】
Since the plant or plant extract of the present invention inhibits the binding of SCF to cell surface c-kit, it has a preventive / therapeutic effect on allergic diseases such as asthma, hay fever, and atopic dermatitis. Can be used as a cosmetic.
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JP3022541B1 (en) * | 1999-01-11 | 2000-03-21 | 株式会社ヘルスサイエンスセンター | External preparation |
JP2000239144A (en) * | 1999-02-22 | 2000-09-05 | Kose Corp | Langerhans cell decrease inhibitor and preparation which contain the inhibitor and is useful for external use for skin |
JP2000247897A (en) * | 1999-02-26 | 2000-09-12 | Ichimaru Pharcos Co Ltd | Cosmetic composition |
JP2001064192A (en) * | 1999-08-25 | 2001-03-13 | Sunstar Inc | Migration inhibitor for langerhans cell and antigen presentation inhibitor |
-
2001
- 2001-04-02 JP JP2001103203A patent/JP3816346B2/en not_active Expired - Fee Related
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