JP2013035808A - Ltbp-4 production promotor - Google Patents
Ltbp-4 production promotor Download PDFInfo
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- JP2013035808A JP2013035808A JP2011175137A JP2011175137A JP2013035808A JP 2013035808 A JP2013035808 A JP 2013035808A JP 2011175137 A JP2011175137 A JP 2011175137A JP 2011175137 A JP2011175137 A JP 2011175137A JP 2013035808 A JP2013035808 A JP 2013035808A
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- ltbp
- algae
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Abstract
Description
本発明は、藻類抽出物または植物抽出物を含有するLTBP-4(Latent TGFβ-binding protein-4)産生促進剤に関する。 The present invention relates to an LTBP-4 (Latent TGFβ-binding protein-4) production promoter containing an algal extract or a plant extract.
LTBP(Latent TGFβ-binding protein;トランスフォーミング増殖因子β結合タンパク質)ファミリーは、血液の凝固に関わる繊維状タンパク質のフィブリンと同じスーパーファミリーに属するタンパク質で、不活性なlatent TGFβを結合することにより、TGFβをマトリックス内に不活性なまま貯蔵し、必要な時にそこからTGFβを動員できるようにする役割がある。 The LTBP (Latent TGFβ-binding protein) family is a protein belonging to the same superfamily as the fibrin protein that is involved in blood coagulation. By binding inactive latent TGFβ, TGFβ Is stored in the matrix in an inactive state, so that TGFβ can be mobilized from there when needed.
このようなタンパク質のうち、LTBP-4について、いくつかの疾患との関連や生体内での機能が知られている。すなわち、LTBP-4の発現と癌進行との間の関連(非特許文献1)の他、LTBP-4が、生体内外における弾性線維形成時のエラスチン沈着の働きを促進することなどが報告されている(特許文献1)。さらに、変異LTBP-4(LTBP4-/-)マウスに肺気腫が見られ、ヒトのCOPDとの関連で発症する遅発性肺気腫の特徴を獲得するという報告もある(非特許文献2および特許文献2)。 Among such proteins, LTBP-4 is known to be associated with several diseases and functions in vivo. That is, in addition to the relationship between LTBP-4 expression and cancer progression (Non-Patent Document 1), it has been reported that LTBP-4 promotes the action of elastin deposition during elastic fiber formation in and outside the body. (Patent Document 1). Furthermore, there are reports that pulmonary emphysema is observed in mutant LTBP-4 (LTBP4-/-) mice and the characteristics of late-onset emphysema that develops in relation to human COPD are acquired (Non-patent Document 2 and Patent Document 2). ).
本発明においては、このような生理活性を有する有用なタンパク質であるLTBP-4の産生促進剤を提供することを目的とする。 An object of the present invention is to provide a production promoter for LTBP-4, which is a useful protein having such physiological activity.
本発明者らは前記課題を解決すべく鋭意検討を重ねた結果、藻類抽出物あるいは植物抽出物に、LTBP-4産生促進能があることを見出し、本発明を完成するに至った。 As a result of intensive studies to solve the above problems, the present inventors have found that an algae extract or a plant extract has an ability to promote LTBP-4 production and have completed the present invention.
即ち本発明は、藻類抽出物または植物抽出物を含有するLTBP-4産生促進剤に関する。 That is, the present invention relates to an LTBP-4 production promoter containing an algal extract or plant extract.
上記LTBP-4産生促進剤は、紅藻類、緑藻類、または褐藻類の抽出物を有効成分として含有し得る。 The LTBP-4 production promoter may contain an extract of red algae, green algae, or brown algae as an active ingredient.
上記紅藻類は、アクロカエティウム目、チノリモ目、ウシケノリ目、ウミゾウメン目、エリスロペルティス目、オオイシソウ目、カクレイト目、サンゴモ目、ダルス目、テングサ目、ロドゴルゴン目、ベニマダラ目、カギノリ目、カギケノリ目、カワモズク目、 スギノリ目、オゴノリ目、マサゴシバリ目、イタニグサ目、およびイギス目に属する藻類からなる群より選択される少なくとも1種であり得る。 The above red algae are Acrocaetium, Chinolemo, Oxenaceae, Cyprididae, Erythropertis, Oenothera, Kakrate, Coralidae, Dulce, Proboscisus, Rhodogorgon, Benimidae, Kaginori, Kagykenori It may be at least one selected from the group consisting of algae belonging to the order of Coleoptera, Cerambycidae, Cerambycidae, Pleurotus, Scorpaenidae, Itabyosa, and Cryptera.
前記緑藻類は、オオヒゲマワリ目、ヨコワミドロ目、サヤミドロ目、ケトフォラ目、ケトペルティス目、クロレラ目、ミクロタムニオン目、トレボウクシア目、カワノリ目、マミエラ目、モノマスティクス目、ドリコマスティクス目、スミレモ目、ウミイカダモ目、クロロデンドロン目、ドリコマスティクス目、ネフロセルミス目、ヒビミドロ目、アオサ目、シオグサ目、モツレグサ目、ミドリゲ目、イワヅタ目、ミル目、ハネモ目、およびカサノリ目に属する藻類からなる群より選択される少なくとも1種であり得る。 The green algae are: Mongolia, Yellow-winged, Sayamidro, Ketophora, Ketopertis, Chlorella, Microtamnion, Trevoxia, Kawanori, Mamiera, Monomastics, Dryomotics, Sumires Selected from the group consisting of algae belonging to the order of: Chlorodendron, Doricomastics, Nephrocellemis, Himidoridae, Aosa, Shiogusa, Ipomoea, Midorige, Iwauta, Milla, Anemone, and Casano There may be at least one.
前記褐藻類は、イソガワラ目、アミジグサ目、ナガマツモ目、カヤモノリ目、ムチモ目、ケヤリモ目、ウルシグサ目、コンブ目、 イシゲ目、およびヒバマタ目に属する藻類からなる群より選択される少なくとも1種であり得る。 The brown algae is at least one selected from the group consisting of algae belonging to the order of Isogawala, Ajigusa, Nagamatsumo, Cyprus, Muchimo, Keyamamo, Urushigusa, Kombu, Ishige, and Hibamata obtain.
前記植物抽出物は、シソ科、マメ科、クロウメモドキ科、マタタビ科、タデ科、ブドウ科、キク科、リンドウ科、ムラサキ科、シロキクラゲ科、ハス科、アオイ科、ミカン科、ショウガ科、アカネ科などに属する植物からなる群より選択される少なくとも1種であり得る。 The plant extract is Lamiaceae, Leguminosae, Crestedaceae, Pteraceae, Tadeidae, Grapeaceae, Asteraceae, Gentianaceae, Muraceae, White Jellyfish, Lotus, Aoi, Citrus, Ginger, Rubiaceae It may be at least one selected from the group consisting of plants belonging to the above.
本発明はまた、上記いずれかのLTBP-4産生促進剤を含有する、LTBP-4産生促進用組成物、に関する。 The present invention also relates to a composition for promoting LTBP-4 production, which contains any of the above LTBP-4 production promoters.
上記LTBP-4産生促進用組成物は、化粧料であり得る。 The composition for promoting LTBP-4 production can be a cosmetic.
上記LTBP-4産生促進用組成物は、食品であり得る。 The LTBP-4 production promoting composition may be a food.
本発明により、藻類または植物の抽出物を有効成分とするLTBP-4産生促進剤が提供される。 The present invention provides an LTBP-4 production promoter containing an algal or plant extract as an active ingredient.
本発明に用いられるLTBP-4は、配列番号1に記載の塩基配列(GenBankアクセッション番号AF051344)若しくはそのオルソログ、またはこれらの変異体(SNP、ハプロタイプを含む)に由来するポリペプチドをいう。LTBP-4のオルソログは特に限定されず、例えば任意の動物、好ましくは哺乳動物に由来するものであり得る。哺乳動物としては、例えばウシ、ヒツジ、ブタ、ヤギ、サル、ウサギ、ラット、ハムスター、モルモット、マウスなどが挙げられる。LTBP-4は分泌タンパク質であり、プロセシングによりシグナル配列が除去され得る。本発明の方法では、LTBP-4としては、シグナル配列が除去されたもの、シグナル配列が除去されていないもののいずれでも使用できるが、シグナル配列が除去されたものが好ましい。また、LTBP-4変異体は、特に限定されず、例えば、配列番号1に示される塩基配列に相補的な配列を有するDNAとストリンジェントな条件でハイブリダイズし且つLTBP-4をコードするDNAに由来するポリペプチド、配列番号1に示される塩基配列がコードするポリペプチドのうち、1つから数個のアミノ酸が欠失、置換、または付加した配列が挙げられる。 LTBP-4 used in the present invention refers to a polypeptide derived from the nucleotide sequence set forth in SEQ ID NO: 1 (GenBank accession number AF051344) or an ortholog thereof, or mutants thereof (including SNP and haplotype). The ortholog of LTBP-4 is not particularly limited, and may be derived from, for example, any animal, preferably a mammal. Examples of mammals include cattle, sheep, pigs, goats, monkeys, rabbits, rats, hamsters, guinea pigs and mice. LTBP-4 is a secreted protein and the signal sequence can be removed by processing. In the method of the present invention, either LTBP-4 from which the signal sequence has been removed or LTBP-4 from which the signal sequence has not been removed can be used, but those from which the signal sequence has been removed are preferred. Further, the LTBP-4 mutant is not particularly limited. For example, the LTBP-4 mutant hybridizes with a DNA having a sequence complementary to the base sequence shown in SEQ ID NO: 1 under stringent conditions and is a DNA encoding LTBP-4. Examples of the polypeptide derived from the polypeptide encoded by the base sequence shown in SEQ ID NO: 1 include a sequence in which one to several amino acids are deleted, substituted, or added.
LTBP-4は、その発現と癌進行との間の関連の他、LTBP-4が、生体内外における弾性線維形成時のエラスチン沈着の働きを促進することなどが報告されている。さらに、肺気腫の治療、ヒトのCOPDとの関連で発症する遅発性肺気腫等への治療への可能性が示唆されている。 In addition to the relationship between its expression and cancer progression, LTBP-4 has been reported to promote the action of elastin deposition during elastic fiber formation in and outside the body. Furthermore, the possibility of treatment of emphysema, and treatment of late-onset emphysema that develops in relation to human COPD has been suggested.
弾性線維は、引き伸ばしても元に戻る「弾性」性質を組織内で担う細胞外線維である。弾性は、体の多くの組織に見られ、肺・動脈・皮膚などのような伸縮性組織に富む組織の機能に必須の性質である。弾性線維は、ミクロフィブリルという線維の周りにエラスチンが沈着・架橋してできると考えられている。エラスチンは、疎水性の強い約750個のアミノ酸からなるタンパク質であり、エラスチン沈着には、その働きを促進するLTBP-4等のタンパク質の関与が知られている。 Elastic fibers are extracellular fibers that take on the “elastic” property in tissues that return to their original state when stretched. Elasticity is found in many tissues of the body and is an essential property for the function of tissues rich in stretchable tissues such as lungs, arteries and skin. Elastic fibers are thought to be formed by depositing and crosslinking elastin around microfibrils. Elastin is a highly hydrophobic protein consisting of about 750 amino acids, and elastin deposition is known to involve proteins such as LTBP-4 that promote its action.
弾性線維は、加齢によって、または炎症細胞から分泌されるプロテアーゼ等の外的要因によって劣化・断裂するため、弾性線維の再生が所望される状態又は疾患が種々存在する。弾性線維の再生が所望される状態又は疾患は、肺気腫、血管損傷、皮膚弛緩症、創傷、弾性線維劣化(例えば、加齢又は紫外線により引き起こされるもの、皮膚のたるみ)、肌荒れ、動脈硬化、大動脈瘤、加齢黄斑変性症、会陰ヘルニア、肛門ヘルニア、脳底動脈瘤、消化管運動障害、褥瘡などである。 Since elastic fibers are deteriorated or broken by aging or by external factors such as protease secreted from inflammatory cells, there are various states or diseases in which regeneration of elastic fibers is desired. Conditions or diseases for which elastic fiber regeneration is desired include emphysema, vascular injury, skin laxity, wounds, elastic fiber degradation (eg, caused by aging or UV rays, skin sagging), rough skin, arteriosclerosis, aorta Aneurysm, age-related macular degeneration, perineal hernia, anal hernia, basilar aneurysm, gastrointestinal motility disorder, pressure ulcer, etc.
本発明は、このような弾性線維形成にも関与しているLTBP-4の産生を促進する物質に関するものであり、このようなLTBP-4産生促進作用が藻類または植物抽出物に見出され、藻類または植物抽出物を有効成分とするLTBP-4産生促進剤が提供される。 The present invention relates to a substance that promotes the production of LTBP-4 that is also involved in the formation of such elastic fibers, and such an LTBP-4 production promoting action is found in algae or plant extracts, An LTBP-4 production promoter comprising an algae or plant extract as an active ingredient is provided.
本発明でいう藻類エキスは、主には、褐藻類、紅藻類または緑藻類から、通常の方法で抽出されるエキスをいう。ここで、紅藻類とは、例えば、ウシケノリ目(ウシケノリ、フノリノウシゲ)、ウミゾウメン目(カモガシラノリ、ヒラガラガラ、フサノリ、ヒラフサノリ、ニセフサノリ、ガラガラ、ベニモヅク、ウミゾウメン)、カクレイト目(ムカデノリ)、サンゴモ目(エゾシコロ、モカサ、オオシコロ)、テングサ目(ユイキリ、シマテングサ、マクサ、オニクサ、オオブサ、オバクサ、ヒラクサ)、ベニマダラ目(ベニマダラ)、カギケノリ目(カギケノリ、タマイタダキ)、スギノリ目(イソダンツウ、ナミイワタケ、アカバ、ミチガエソウ、ハナフノリ、フクロフノリ、カイノリ、シキンノリ、スギノリ、コトジツノマタ、オオバツノマタ、ツノマタ、イボツノマタ、アカバギンナンソウ、イトフノリ、ヒロハノカクレイト、キントキ、フシキントキ、ムカデノリ、ニクムカデ、ウツロムカデ、ヒトツマツ、カタノリ、ナガキントキ、タンバノリ、フダラク、ヒラムカデ、ヒラキントキ、スジムカデ、オオバキントキ、ヒヂリメン、ツルツル、マツノリ、キョウノヒモ、マタボウ、コメノリ、ツノムカデ、トサカマツ、イバラノリ、コヒモイバラ、ホシガタイバラ、スジイバラノリ、カヅノイバラ、カギイバラノリ、サイダイバラ、タチイバラ、ヒロハノトサカモドキ、ホソバノトサカモドキ、ウスギヌ、イカノアシ、エツキイワノカワ、キジノオ、サイミ、オキツノリ、ハリガネ、ハスジグサ、マキユカリ、ユカリ、ホソバナミノハナ、ナミノハナ、アツバノリ、ベニスナゴ、トサカノリ、ミリン)、オゴノリ目(シラモ、ミゾオゴノリ、ベニオゴノリ、フシクレノリ、カバノリ、オゴノリ、マサゴシバリ目(ワツナギソウ、イソマツ、カイメンソウ、タオヤギソウ、フシツナギ、コスジフシツナギ、フクロツナギ、ニセイバラノリ、マサゴシバリ)、イギス目(フトイギス、エゴノリ、トゲイギス、ハネイギス、イギス、ハリイギス、ケイギス、カザシグサ、キヌイトカザシグサ、ニクサエダ、サエダ、ベニヒバ、ナガウブゲグサ、ランゲリア、ヤレウスバノリ、スジウスバノリ、カギウスバノリ、タチウスベニ、ハスジギヌ、トゲノリ、ツクシホウズキ、ベンテンモ、ユナ、ヤナギノリ、マクリ、ケハネグサ、クロソゾ、ニッポンソゾ、ウラソゾ、ミツデソゾ、パピラソゾ、ハネソゾ、マギレソゾ、コブソゾ、ジャバラノリ、フジマツモ、ショウジョウケノリ、イソムラサキ、コザネモ)、アクロカエティウム目、チノリモ目、エリスロペルティス目、オオイシソウ目、ダルス目、ロドゴルゴン目、カギノリ目、カワモズク目、イタニグサ目、などに属する藻類をいう。 The algal extract referred to in the present invention mainly refers to an extract extracted from brown algae, red algae or green algae by a usual method. Here, the red algae is, for example, the order of the genus Oleoptera (Oxenaceae, Funolino bogei), Ezozoenidae (Anemone quercivorus, Pleurotus serrata, Fusanori, Hirasanori, Nisehusanori, Ragara, Benimo zouk, Ezozoen) ) , Kainori, Chikinori, Suginori, Kotojitsu no Mata, Oobatsu No Mata, Tsuno Mata, Ibottsu No Mata, Akabaginanso, Atomugi Nori, Hirohanokakrate, Kintoki, Fushi Totoki, Centipede, Nikumukade, Utsuromukade, Hitomitsu, Katanari, Nagakintoki, Tambanori, Fudarak, Hiramkade, Hirakintoki, Sumidakade, Obakintoki, Hijirimen, Tsurutsuru, Matsunori, Masuhito , Coconut rose, key swordfish, seidai rose, tachiibara, hirohanotosakamodoki, hosobanotosakamodoki, usuginu, ikanoashi, etsukiiwakawa, kijinoo, saimi, okitsunori, harigane, mizusuna ), Pleurotus (Shiramo, Mizoogonori, Beniogonori, Fusiclenori, Birch Steller's sea bream, scorpionate (Villus serrata, Cyprinus, Caenopus, Taoyagisou, Fushitsunagi, Kosushifushitsunagi, Fukutsunagi, Nisebara Nori, Masagoshibari), Locust (Futagigis, Egonori, Togegisu, Hagisugi, Hagizugi Saeda, Benihiba, Nagabugugusa, Langeria, Yaleus Banori, Susius Banori, Kagios Banori, Tachius Beni, Hasujiginu, Togenori, Tsukubiwazo, Benzomo, Yuna, Yanaginori, Makuri, Sozozosozode , Fujimatsumo, Drosophila chinensis, Isomurasaki, Kozanemo), Acrocaetium, Chi It refers to algae belonging to the order of Norimo, Erythropertis, Euphorbiaceae, Dulce, Rhodogorgon, Kaginori, Kawamoku, Itabusa.
緑藻類とは、ヒビミドロ目 (シワランソウモドキ、ランソウモドキ)、アオサ目(ナガアオサ、ボタンアオサ、リボンアオサ、ボウアオノリ、ウスバアオノリ、アナアオサ、アミアオサ、ヤブレグサ)、シオグサ目(ウキオリソウ、タルガタジュズモ、ホソジュズモ、ウスイロジュズモ、フトジュズモ、カイゴロモ、オオシオグサ、カタシオグサ、アサミドリシオグサ、チャシオグサ)、モツレグサ目(シリオミドロ)、ミドリゲ目(アオモグサ、マガタマモ、キッコウグサ、タマバロニア、タマゴバロニア、オオバロニア)、イワヅタ目(ヘライワヅタ、フジノハヅタ、フサイワヅタ、クサビガタハウチワ、イトゲノマユハキ、マユハキモ、ヒメイチョウ)、ミル目(ナンバンハイミル、ヒゲミル、ネザシミル、サキブトミル、ナガミル、ミル、モツレミル、ヒラミル、タマミル、コブシミル、クロミル)、ハネモ目(カタハノハネモ、オオハネモ、ナンカイハネモ)、カサノリ目(ミズタマ、フデノホ、リュウキュウガサ、カサノリ、イソスギナ)、オオヒゲマワリ目、ヨコワミドロ目、サヤミドロ目、ケトフォラ目、ケトペルティス目、クロレラ目、ミクロタムニオン目、トレボウクシア目、カワノリ目、マミエラ目、モノマスティクス目、ドリコマスティクス目、スミレモ目、ウミイカダモ目、クロロデンドロン目、ドリコマスティクス目、ネフロセルミス目、などに属する藻類をいう。 Green algae are: Hibidoro (Siwaransomodoki, Ranodomodoki), Aosasa (Nagaaosa, Button Aosa, Ribbon Aosa, Bowuaonori, Usubaaoori, Anaaaosa, Amiaosa, Yabregusa) Cairomomo, Giant Pterosa, Caterpillar, Assamirisiogusa, Chasiogusa), Ipomoea (Siriomidro), Midorigea (Aomogusa, Magamata, Duckweed), Tamavalonia, Tamagobaronia, Ovalonia, Iwauta, Chileoptera Itogenomayuhaki, Mayuhakimo, Hime butterfly), mil eyes (Nanban High Mill, Higemil, Nezashimir, Sakibutomi, Nagamil, Mill, Motelle Mill, Hiramil, Tamil, Kobushimiru, Cromil), Anemone (Katahanohanemo, Ohananemo, Nankaihanemo), Casanori (Mizutama, Fudenoho, Ryukyugasa, Casanori, Isosugina), Ohigemawara eyes, Yokomado , Ketopertis, Chlorella, Microtamnion, Trevoxia, Kawanori, Mamiera, Monomastics, Dreamomastic, Sumiremo, Umikadamamo, Chlorodendron, Dreamomastic, Nephrothermis, etc. The algae to which it belongs.
褐藻類とは、イソガワラ目(マツモ、キンイロハンモン)、アミジグサ目(エゾヤハズ、ヤハズグサ、ヘラヤハズ、シワヤハズ、アミジグサ、イトアミジ、フクリンアミジ、フタエオオギ、サナダグサ、ウミウチワ、コナウミウチワ、アツバコモングサ、コモングサ、シマオオギ)、ナガマツモ目(クロモ、フトモヅク、ネバリモ、シワノカワ)、カヤモノリ目(ワタモ、ホソクビワタモ、フクロノリ、カゴメノリ、イワヒゲ、ハバノリ、セイヨウハバノリ、カヤモドキ、カヤモノリ)、ムチモ目(ムチモ)、ケヤリモ目(イチメガサ)、ウルシグサ目(ウルシグサ、タバコグサ)、コンブ目(ワカメ、ツルモ、スジメ、カジメ、クロメ、アラメ、マコンブ、ホソメコンブ)、イシゲ目(イシゲ、イロロ)、ヒバマタ目(ウガノモク、ヤバネモク、ジョロモク、スナビキモク、フシスジモク、ホンダワラ、ヒジキ、オオバノコギリモク、イソモク、アカモク、ノコギリモク、タマハハキモク、ナラサモ、ヒラネジモク、ヤツマタモク、マメタワラ、オオバモク、ウミトラノオ、アズマネジモク、ヨレモクモドキ、エンドウモク、ラッパモク)などをいう。 Brown algae are: Isogawala (Matsumo, Kinirohammon), Amygusa (Ezoyahaz, Yahazugusa, Herayahaz, Shiwayahaz, Ajigusa, Itoamiji, Fukurimamiji, Phaeosagi, Sanadagusa, Umichiwasa, Osamu Osa (Chromo, Futomoku, Nebarimo, Shiwanokawa), Crayfish eyes (Watamo, Hosookuwatamamo, Fukuronori, Kagomenori, Iwahige, Havanori, Habanori, Kayamo-doki, Kayanori), Whip-eye (Polypoda), Shigusa-Ru (Ishigusa-guru) Tobacco rush), Kombu eyes (Wakame, Tsurumo, Sujime, Kajime, Kurome, Arame, Macombu, Hosome Kombu), Shigego (Ishige, Iroro), Hibamata (Uganomok, Yaban) Say click, Joromoku, Sunabikimoku, Fushisujimoku, Sargassum, hijiki, Oba saw-Mok, Isomoku, akamoku, Nokogirimoku, Tamahahakimoku, Narasamo, Hiranejimoku, Yatsumatamoku, Mametawara, Oobamoku, Umitoranoo, Azumanejimoku, Yoremokumodoki, Endoumoku, Rappamoku), and the like.
本発明においては、このようなLTBP-4の産生を促進する作用が、藻類または植物の抽出物に見出された。藻類または植物からの抽出物は、1種を単独または2種以上を組み合わせて使用しても良い。このような藻類または植物としては、限定されず、また、抽出物とは、LTBP-4の産生促進作用が損なわれない限り、藻類または植物からの水可溶性成分、酸可溶性成分、アルカリ可溶性成分、有機溶媒可溶性成分などをいう。 In the present invention, the effect of promoting the production of LTBP-4 was found in algae or plant extracts. Extracts from algae or plants may be used singly or in combination of two or more. Such an algae or plant is not limited, and an extract is a water-soluble component, an acid-soluble component, an alkali-soluble component from an algae or a plant, as long as the production promoting action of LTBP-4 is not impaired. An organic solvent soluble component.
本発明においては、このようなLTBP-4の産生を促進する作用を有する成分としては、藻類および植物から抽出されるのもあれば特に限定はない。例えば、褐藻類はフコイダンが、紅藻類はカラギーナンおよび/またはケイ素が、緑藻類はクロロフィルαおよび/またはβ等が含まれていても良い。 In the present invention, the component having the action of promoting the production of LTBP-4 is not particularly limited as long as it can be extracted from algae and plants. For example, brown algae may contain fucoidan, red algae may contain carrageenan and / or silicon, and green algae may contain chlorophyll α and / or β.
本発明におけるLTBP-4産生促進作用は、植物抽出物にも見出される。本明細書における「植物抽出物」には、植物から抽出した成分の他、植物から抽出した成分と例えば乳酸菌を共存させて得られるような植物抽出分発酵液も含まれる。植物としては、例えば、アオイ科、アカネ科、アケビ科、アブラナ科、アヤメ科、イネ科、イラクサ科、イワウメ科、 イワデンダ科、ウコギ科、ウマノスズクサ科、ウリ科、オオバコ科、オトギリソウ科、キク科、キキョウ科、キンポウゲ科、クスノキ科、クロウメモドキ科、クワ科、ケシ科、ゴマノハグサ科、コバノイシカグマ科、サトイモ科、サクラソウ科、シソ科、シラネアオイ科、シロキクラゲ科、ショウガ科、ジンチョウゲ科、スイカズラ科、スイレン科、スミレ科、セリ科、ゼンマイ科、タデ科、ツゲ科、ツツジ科、ツユクサ科、ドクダミ科、トチノキ科、ナス科、ナデシコ科、ハス科、バラ科、ヒガンバナ科、ヒルガオ科、フウロウソウ科、ブドウ科、ブナ科、ベンケイソウ科、ボタン科、マタタビ科、マツ科、マメ科、マンサク科、ミカン科、ミズキ科、ミツガシワ科、ムラサキ科、メギ科、ヤマノイモ科、ユキノシタ科、ユリ科、ラン科、リンドウ科、レンプクソウ科、ノウゼンカズラ科などに属する植物が挙げられる。 The LTBP-4 production promoting action in the present invention is also found in plant extracts. The “plant extract” in the present specification includes, in addition to components extracted from plants, plant extract fermented liquids obtained by coexisting components extracted from plants and, for example, lactic acid bacteria. Plants include, for example, mallow, rhododendron, echidaceae, cruciferous, iridaceae, gramineae, nettleaceae, cranaceae, cichlidae, araliaceae, urchinaceae, cucurbitaceae, plantain, hypericaceae, asteraceae , Asteraceae, buttercup, camphoraceae, buckthorn, mulberry, poppy, scorpionaceae, scorpionaceae, araceae, primrose, perilla, shiraneae, ginger, ginger, hydrangea, hydrangea Family, violet family, celery family, spring mackerel family, aceae family, boxwood family, azalea family, camellia family, dodami family, cypress family, eggplant family, dianthus family, lotus family, rose family, bumblebee family, convolvulaceae family, cypress family, Grapeaceae, beech, diatomaceae, button, matabidae, pine, legume, man Examples include plants belonging to the family Cactaceae, Citrusaceae, Citricaceae, Sphagnumaceae, Purpleaceae, Barberry, Yamanoidae, Saxifragaceae, Lilyaceae, Orchidaceae, Gentianaceae, Lempoxaceae, Nodusaceae.
このような植物抽出物は、特に限定されないが、シソ科のオウゴンエキス、ノウゼンカズラ科のタベブイアインペチギノサ、マメ科のカンゾウ根エキス、クロウメモドキ科のナツメ果実エキス、マタタビ科のキウイ種子エキス、タデ科のアイ葉、キク科のアーチチョーク葉エキス、タベブイアインペチギノサ樹皮エキス、リンドウ科のゲンチアナ根エキス、ムラサキ科のシコンエキス、ムラサキ科のムラサキ根エキス、シロキクラゲ科のシロキクラゲ抽出物または多糖体含有シロキクラゲ抽出物、アオイ科のバオバブ葉エキスまたは加水分解バオバブ葉エキス、ミカン科のトウヒエキスまたはビターオレンジ果皮エキス、ミカン科のトウヒエキス/ビターオレンジ果皮エキス、ショウガ科の月桃葉エキス、およびアカネ科のアセンヤクエキスからなる群より選択される1種又は2種以上である。なお、上記植物の抽出物が商業的に入手できるものである場合には、簡便には、当該市販の植物抽出物を用いることもできる。 Such plant extracts are not particularly limited, but include osconus urgonum extract, genus saffronaceae tabebuia impetiginosa, leguminous licorice root extract, buckthorn jujube fruit extract, matabidae kiwi seed extract, laceaceae Eye Leaf, Compositae Arch Chalk Leaf Extract, Tabebuia Impetiginosa Bark Extract, Gentian Gentiana Root Extract, Muraceae Shikon Extract, Murasaki Murasaki Root Extract, White Jellyfish Extract or Polysaccharide-Containing White Jellyfish Extract , Aceraceae Baobab leaf extract or hydrolyzed Baobab leaf extract, Citrus spruce or bitter orange peel extract, Citrus spruce extract / bitter orange peel extract, Ginger department moon peach leaf extract, and Acaenaceae asenyaku Extract 1 type or 2 types or more selected from the group which consists of. In addition, when the said plant extract is what can be obtained commercially, the said commercially available plant extract can also be used simply.
以下に、本発明の内容を詳細に説明する。 The contents of the present invention will be described in detail below.
藻類または植物抽出物は、天然物からの抽出液の粗生成物をそのまま用いても、また、粗生成物を分離精製したものを用いてもよい。 As the algae or plant extract, a crude product of an extract from a natural product may be used as it is, or a product obtained by separating and purifying a crude product may be used.
藻類または植物の抽出物を得る方法としては、特に限定されず、通常の抽出法が採用される。粗抽出液は、例えば全藻または植物の全体あるいはそれらの特定の部位(根、茎、幹、樹皮、幼芽、葉、花、果実及び種子等の植物の一部)を裁断機・スライサー・カッター・ピーラーなどで細断したものを、水、熱水あるいは有機溶媒を単独あるいは組み合わせて得られる溶媒を用いて抽出することにより得ることができる。有機溶媒としては、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−ブタノール等のアルコール類、酢酸メチル、酢酸エチル等のエステル類、アセトン等のケトン類、ホルムアルデヒド、ジメチルスルホキシドなどを挙げることができる。 The method for obtaining the algae or plant extract is not particularly limited, and a normal extraction method is employed. The crude extract is a cutting machine, slicer, etc., for example, whole algae or whole plants or specific parts thereof (roots, stems, stems, bark, buds, leaves, flowers, fruits, seeds, etc.). What is shredded by a cutter / peeler or the like can be obtained by extracting with water, hot water, or a solvent obtained by combining organic solvents alone or in combination. Examples of the organic solvent include alcohols such as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol and 2-butanol, esters such as methyl acetate and ethyl acetate, ketones such as acetone, formaldehyde, dimethyl sulfoxide and the like. Can be mentioned.
また、溶媒は、室温又は常温の溶媒を用いることもできるが、加熱溶媒や熱溶媒(溶媒を沸点付近に加熱したもの)を用いることもできる。 In addition, a solvent at room temperature or room temperature can be used as the solvent, but a heating solvent or a thermal solvent (a solvent heated near the boiling point) can also be used.
さらに、酸、アルカリの条件下で抽出する事もできる。酸としては、塩酸、硫酸、硝酸、リン酸などの無機酸、又は、酢酸、クエン酸、シュウ酸、コハク酸、ギ酸、プロピオン酸などの有機酸が挙げられる。アルカリとしては、アルカリ無機塩が好ましく用いられ、水酸化カリウム、水酸化ナトリウム等を例示できる。 Furthermore, extraction can be performed under acid and alkali conditions. Examples of the acid include inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid, and organic acids such as acetic acid, citric acid, oxalic acid, succinic acid, formic acid, and propionic acid. As the alkali, an alkali inorganic salt is preferably used, and examples thereof include potassium hydroxide and sodium hydroxide.
また、凍結乾燥粉末化法、高圧法や超高圧法を利用する事も可能である。高圧法においては120℃、4時間の高圧加熱処理することにより、当該藻類または植物中の細胞が加圧され破砕される。 It is also possible to use a freeze-drying powdering method, a high pressure method or an ultrahigh pressure method. In the high-pressure method, the cells in the algae or plant are pressurized and crushed by high-pressure heat treatment at 120 ° C. for 4 hours.
粗抽出物の抽出条件の一例を挙げる。抽出溶媒が有機溶媒の場合、抽出温度は特に制限はないが、好ましくは1〜100℃、より好ましくは20〜100℃の範囲で、1〜24時間、好ましくは1時間〜10時間撹拌しながら行う。また、抽出溶媒が水の場合、抽出時の温度は、溶媒の沸点以下であればよく特に限定されないが、加圧熱水による抽出方法を利用する場合もあるため、約0〜200℃が好ましく、約20〜130℃がより好ましい。また、反応時間も特に限定するものではないが、好ましくは数秒間〜数日間に設定すれば良く、更に好ましくはい30分〜6時間に設定すれば良い。 An example of the extraction conditions for the crude extract is given. When the extraction solvent is an organic solvent, the extraction temperature is not particularly limited, but preferably 1 to 100 ° C, more preferably 20 to 100 ° C, with stirring for 1 to 24 hours, preferably 1 hour to 10 hours. Do. Further, when the extraction solvent is water, the temperature at the time of extraction is not particularly limited as long as it is equal to or lower than the boiling point of the solvent. However, since an extraction method using pressurized hot water may be used, about 0 to 200 ° C is preferable. About 20 to 130 ° C. is more preferable. Also, the reaction time is not particularly limited, but it may be set to several seconds to several days, more preferably 30 minutes to 6 hours.
反応温度及び反応時間は、溶媒の種類と濃度により適宜選択すれば良い。 What is necessary is just to select reaction temperature and reaction time suitably according to the kind and density | concentration of a solvent.
このように抽出されたものであれば、LTBP‐4産生促進能を有する成分を十分に抽出することができる。 If it is extracted in this way, the component which has LTBP-4 production promotion ability can fully be extracted.
得られた抽出物を遠心分離してその上清を、抽出液として得る。かかる抽出液に水可溶性、酸性可溶性、アルカリ可溶性、あるいは有機溶媒可溶性の多糖画分が含まれる。 The obtained extract is centrifuged to obtain the supernatant as an extract. Such an extract contains a water-soluble, acidic-soluble, alkali-soluble, or organic solvent-soluble polysaccharide fraction.
抽出物の粗生成物から藻類または植物抽出物を分離精製する方法としては、例えば、塩化セチルピリジニウムなどの4級アンモニウム塩を添加し、藻類または植物抽出物を沈殿させ、アルコール溶液で洗浄する方法やDEAEセルロースなどの陰イオン交換樹脂などを用いたクロマトグラフィーを用いる方法もある。 As a method for separating and purifying an algae or plant extract from a crude product of the extract, for example, a method of adding a quaternary ammonium salt such as cetylpyridinium chloride to precipitate the algae or plant extract and washing with an alcohol solution There is also a method using chromatography using an anion exchange resin such as DEAE cellulose.
このように抽出、精製した藻類または植物抽出物溶液は、そのまま用いることもできる他、稀釈液で希釈したり、濃縮してエキスとして用いることもでき、さらには、凍結乾燥などにより乾燥粉末物としたり、ペースト状に調製して用いることもできる。 The algae or plant extract solution thus extracted and purified can be used as it is, or can be diluted with a diluting solution or concentrated to be used as an extract. Or it can be prepared in the form of a paste and used.
本発明の藻類または植物抽出物を有効成分とするLTBP-4産生促進剤は、LTBP-4の発現量が低下していることを原因とする疾患の治療に用いることができる。例えば、癌、肺気腫の治療、ヒトのCOPDとの関連で発症する遅発性肺気腫等への治療への可能性がある。また、LTBP-4産生が関わっているとされる弾性線維形成に関わる疾患の治療にも有用である。弾性線維形成に関わる疾患とは、弾性線維の再生が所望される状態又は疾患のことであり、例えば、肺気腫、血管損傷、皮膚弛緩症、創傷、弾性線維劣化(例えば、加齢又は紫外線により引き起こされるもの)、肌荒れ、動脈硬化、大動脈瘤、加齢黄斑変性症、会陰ヘルニア、肛門ヘルニア、脳底動脈瘤、消化管運動障害、褥瘡などを指す。さらには、皮膚のたるみやしわの予防や改善といった美容目的のためにも有用である。また、本発明のLTBP-4産生促進剤は、弾性線維の再生能を有する細胞の培養用試薬などとしても有用である。さらに、人工皮膚、皮膚モデル、人工血管などの人工組織の作製に役立つ。さらに、十分解明されているとはいえないLTBP-4自体の研究への応用も考えられ、非常に広い範囲で有用であると考えられる。 The LTBP-4 production promoter comprising the algae or plant extract of the present invention as an active ingredient can be used for the treatment of diseases caused by a decrease in the expression level of LTBP-4. For example, there are possibilities for treatment of cancer, emphysema, and late-onset emphysema that develops in relation to human COPD. It is also useful for the treatment of diseases related to the formation of elastic fibers that are thought to involve LTBP-4 production. A disease associated with elastic fiber formation is a condition or disease in which elastic fiber regeneration is desired, such as pulmonary emphysema, vascular injury, skin laxity, wounds, elastic fiber degradation (eg, caused by aging or ultraviolet radiation). ), Rough skin, arteriosclerosis, aortic aneurysm, age-related macular degeneration, perineal hernia, anal hernia, basilar aneurysm, gastrointestinal motility disorder, pressure ulcer, etc. Furthermore, it is useful for cosmetic purposes such as prevention and improvement of skin sagging and wrinkles. The LTBP-4 production promoter of the present invention is also useful as a cell culture reagent having the ability to regenerate elastic fibers. Furthermore, it is useful for producing artificial tissues such as artificial skin, skin models, and artificial blood vessels. Furthermore, LTBP-4 itself, which has not been fully elucidated, can be applied to research, and is considered useful in a very wide range.
本発明のLTBP-4産生促進剤は、医薬品、医薬部外品、食品、化粧品などとして幅広く利用可能な任意の製剤または組成物の形態で提供され得るが、好ましくは、機能性食品または化粧品として利用可能な製剤または組成物である。 The LTBP-4 production promoter of the present invention can be provided in the form of any preparation or composition that can be widely used as a pharmaceutical, quasi-drug, food, cosmetic, etc., but preferably as a functional food or cosmetic. Available formulation or composition.
本発明のLTBP-4産生促進剤を含む組成物が、医薬品、医薬部外品、食品、化粧品のいずれの形態を取る場合でも、藻類または植物の抽出物の含有量は、乾燥重量に換算して、例えば約0.000001〜99重量%とすればよく、より好ましくは約0.00001〜30重量%、更に好ましくは約0.0001〜10重量%、特に好ましくは0.001〜5重量%である。上記範囲であれば、医薬品、医薬部外品、食品、化粧品の使用で十分なLTBP-4産生促進作用がみられる。 Regardless of whether the composition containing the LTBP-4 production promoter of the present invention takes any form of pharmaceuticals, quasi-drugs, foods, and cosmetics, the algae or plant extract content is converted to dry weight. For example, it may be about 0.000001 to 99% by weight, more preferably about 0.00001 to 30% by weight, still more preferably about 0.0001 to 10% by weight, particularly preferably 0.001 to 5% by weight. It is. If it is the said range, sufficient LTBP-4 production promotion effect is seen by use of a pharmaceutical, a quasi-drug, a foodstuff, and cosmetics.
本発明を医薬品または健康食品の用途に用いる場合の藻類または植物の抽出物の投与量は、乾燥重量にして、成人1日当り0.0001〜5000mg/kgとすることができる。上記範囲であれば、LTBP‐4産生を必要とする疾患または状態の改善や予防が可能となる。 When the present invention is used for pharmaceuticals or health foods, the dose of algae or plant extract can be 0.0001 to 5000 mg / kg per day for adults in dry weight. If it is the said range, the improvement or prevention of the disease or condition which requires LTBP-4 production is attained.
皮膚外用剤形態 Skin topical form
本発明の藻類または植物の抽出物(粗抽出物または精製物)を有効成分とするLTBP-4産生促進剤は、化粧料又は医薬部外品の外用剤形態として公知の形態を採ることができる。このような形態として、例えば、液剤、懸濁剤、乳剤、クリーム剤、軟膏剤、ゲル剤、リニメント剤、ローション剤、エアゾール剤、パウダー剤、パップ剤、不織布等のシートに薬液を含浸させたシート剤、リップスティックのようなスティック剤などが挙げられる。中でも、好ましくは、液剤、懸濁剤、乳剤、クリーム剤、軟膏剤、ゲル剤、ローション剤、ゲル剤の形態で用いられる。かかる形態とすることにより、弾性線維形成効果を十分に発揮することができる。皮膚外用剤には、藻類または植物の抽出物に加えて、本発明の効果を損なわない範囲で、外皮用剤(化粧料、医薬部外品、医薬品)に添加される公知の添加剤、例えば、界面活性剤、油分、アルコール類、保湿剤、増粘剤、保存剤(防腐剤)、酸化防止剤、キレート剤、pH調整剤、安定化剤、分散剤、香料、着色剤、色素、パール光沢付与剤、血行促進成分、保湿成分、紫外線吸収成分、紫外線散乱成分、洗浄成分、抗菌成分、収斂成分、ビタミン類、アミノ酸類、老化防止成分、抗炎症成分、美白成分、角質柔軟成分、細胞賦活化成分、水等を配合することができる。添加剤は、1種を単独で、又は2種以上を組み合わせて使用できる。 The LTBP-4 production promoter comprising the algae or plant extract (crude extract or purified product) of the present invention as an active ingredient can take a form known as an external preparation form of cosmetics or quasi drugs. . As such forms, for example, liquids, suspensions, emulsions, creams, ointments, gels, liniments, lotions, aerosols, powders, poultices, nonwoven fabrics and the like were impregnated with chemicals. Examples thereof include a sheet agent and a stick agent such as a lipstick. Among these, the liquid, suspension, emulsion, cream, ointment, gel, lotion, and gel are preferably used. By setting it as this form, the elastic fiber formation effect can fully be exhibited. For skin external preparations, in addition to algal or plant extracts, known additives that are added to skin external preparations (cosmetics, quasi-drugs, pharmaceuticals) within a range that does not impair the effects of the present invention, for example, , Surfactants, oils, alcohols, humectants, thickeners, preservatives (preservatives), antioxidants, chelating agents, pH adjusters, stabilizers, dispersants, fragrances, colorants, pigments, pearls Gloss imparting agent, blood circulation promoting component, moisturizing component, UV absorbing component, UV scattering component, cleaning component, antibacterial component, astringent component, vitamins, amino acids, anti-aging component, anti-inflammatory component, whitening component, keratin softening component, cell An activation component, water, etc. can be mix | blended. An additive can be used individually by 1 type or in combination of 2 or more types.
また、化粧料組成物の具体的な用途は特に限定されず、化粧水、乳液、クリーム、美容液、日焼け止め用化粧料、パック、ハンドクリーム、ボディローション、ボディークリームのような基礎化粧料;洗顔料、メイク落とし、ボディーシャンプー、シャンプー、リンスのような洗浄用化粧料;ファンデーション、化粧下地、リップクリーム、口紅、チークカラーのようなメークアップ化粧料;入浴剤などが挙げられる。 In addition, the specific use of the cosmetic composition is not particularly limited, and basic cosmetics such as lotion, milky lotion, cream, cosmetic liquid, sunscreen cosmetics, packs, hand creams, body lotions, body creams; Cosmetics for cleaning such as face wash, makeup remover, body shampoo, shampoo, rinse; makeup cosmetics such as foundation, makeup base, lip balm, lipstick, teak color;
<基剤又は担体>
基剤又は担体としては、流動パラフィン、スクワラン、ゲル化炭化水素(プラスチベースなど)、オゾケライト、α−オレフィンオリゴマー、軽質流動パラフィンのような炭化水素;メチルポリシロキサン、架橋型メチルポリシロキサン、高重合メチルポリシロキサン、環状シリコーン、アルキル変性シリコーン、架橋型アルキル変性シリコーン、アミノ変性シリコーン、ポリエーテル変性シリコーン、ポリグリセリン変性シリコーン、架橋型ポリエーテル変性シリコーン、架橋型アルキルポリエーテル変性シリコーン、シリコーン・アルキル鎖共変性ポリエーテル変性シリコーン、シリコーン・アルキル鎖共変性ポリグリセリン変性シリコーン、ポリエーテル変性分岐シリコーン、ポリグリセリン変性分岐シリコーン、アクリルシリコン、フェニル変性シリコーン、シリコーンレジンのようなシリコーン油;エチルセルロース、ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロースのようなセルロース誘導体;ポリビニルピロリドン;カラギーナン;ポリビニルブチラート;ポリエチレングリコール;ジオキサン;ブチレングリコールアジピン酸ポリエステル;ミリスチン酸イソプロピル、ミリスチン酸オクチルドデシル、パルミチン酸イソプロピル、パルミチン酸セチル、イソノナン酸イソノニル、テトラ2−エチルヘキサン酸ペンタエリスリットのようなエステル類;デキストリン、マルトデキストリンのような多糖類;エタノール、イソプロパノールのような低級アルコール;エチレングリコールモノメチルエーテル、エチレングリコールモノエチルエーテル、エチレングリコールモノプロピルエーテル、ジエチレングリコールモノメチルエーテル、ジエチレングリコールモノエチルエーテル、ジエチレングリコールモノプロピルエーテル、ジエチレングリコールモノブチルエーテル、プロピレングリコールモノエチルエーテル、プロピレングリコールモノプロピルエーテル、ジプロピレングリコールモノエチルエーテル、ジプロピレングリコールモノプロピルエーテルのようなグリコールエーテル;ポリエチレングリコール、プロピレングリコール、1,3-ブチレングリコール、グリセリン、イソプレングリコールなどの多価アルコール;水などの水系基剤などが挙げられる。
<Base or carrier>
Bases or carriers include liquid paraffin, squalane, gelled hydrocarbons (such as plastibase), ozokerite, α-olefin oligomers, hydrocarbons such as light liquid paraffin; methyl polysiloxane, cross-linked methyl polysiloxane, highly polymerized methyl Polysiloxane, Cyclic silicone, Alkyl-modified silicone, Cross-linked alkyl-modified silicone, Amino-modified silicone, Polyether-modified silicone, Polyglycerin-modified silicone, Cross-linked polyether-modified silicone, Cross-linked alkyl polyether-modified silicone, Silicone / alkyl chain copolymer Modified polyether-modified silicone, silicone / alkyl chain co-modified polyglycerin-modified silicone, polyether-modified branched silicone, polyglycerin-modified branched silicone, acrylic silicone, Silicone oils such as phenyl-modified silicones and silicone resins; cellulose derivatives such as ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose; polyvinylpyrrolidone; carrageenan; polyvinyl butyrate; polyethylene glycol; dioxane; butylene glycol adipate polyester; isopropyl myristate , Esters such as octyldodecyl myristate, isopropyl palmitate, cetyl palmitate, isononyl isononanoate, pentaerythritol tetra-2-ethylhexanoate; polysaccharides such as dextrin and maltodextrin; lower levels such as ethanol and isopropanol Alcohol; ethylene glycol monomethyl ether, ethylene glycol monoethyl ether , Ethylene glycol monopropyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monopropyl ether, diethylene glycol monobutyl ether, propylene glycol monoethyl ether, propylene glycol monopropyl ether, dipropylene glycol monoethyl ether, dipropylene glycol monopropyl Examples include glycol ethers such as ethers; polyhydric alcohols such as polyethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin, and isoprene glycol; and aqueous bases such as water.
基剤又は担体は、1種を単独で、又は2種以上を組み合わせて使用できる。 A base or a support | carrier can be used individually by 1 type or in combination of 2 or more types.
<添加剤>
本発明の外用組成物に含まれ得る酸化防止剤としては、ジブチルヒドロキシトルエン、ブチルヒドロキシアニソール、ソルビン酸、亜硫酸ナトリウム、アスコルビン酸、エリソルビン酸、L−システイン塩酸塩などが挙げられる。
<Additives>
Examples of the antioxidant that can be contained in the composition for external use of the present invention include dibutylhydroxytoluene, butylhydroxyanisole, sorbic acid, sodium sulfite, ascorbic acid, erythorbic acid, L-cysteine hydrochloride and the like.
界面活性剤としては、例えば、ソルビタンモノイソステアレート、ソルビタンモノラウレート、ソルビタンモノパルミテート、ソルビタンモノステアレート、ペンタ−2−エチルヘキシル酸ジグリセロールソルビタン、テトラ−2−エチルヘキシル酸ジグリセロールソルビタンのようなソルビタン脂肪酸エステル類;モノステアリン酸プロピレングリコールのようなプロピレングリコール脂肪酸エステル類;ポリオキシエチレン硬化ヒマシ油40(HCO−40)、ポリオキシエチレン硬化ヒマシ油50(HCO−50)、ポリオキシエチレン硬化ヒマシ油60(HCO−60)、ポリオキシエチレン硬化ヒマシ油80などの硬化ヒマシ油誘導体;モノラウリル酸ポリオキシエチレン(20)ソルビタン(ポリソルベート20)、モノステアリン酸ポリオキシエチレン(20)ソルビタン(ポリソルベート60)、モノオレイン酸ポリオキシエチレン(20)ソルビタン(ポリソルベート80)、イソステアリン酸ポリオキシエチレン(20)ソルビタンのようなポリオキシエチレンソルビタン脂肪酸エステル類;ポリオキシエチレンモノヤシ油脂肪酸グリセリル;グリセリンアルキルエーテル;アルキルグルコシド;ポリオキシエチレンセチルエーテルのようなポリオキシアルキレンアルキルエーテル;ステアリルアミン、オレイルアミンのようなアミン類;ポリオキシエチレン・メチルポリシロキサン共重合体、ラウリルPEG−9ポリジメチルシロキシエチルジメチコン、PEG−9ポリジメチルシロキシエチルジメチコンのようなシリコーン系界面活性剤などが挙げられる。 Examples of the surfactant include sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, diglycerol sorbitan penta-2-ethylhexylate, and diglycerol sorbitan tetra-2-ethylhexylate. Sorbitan fatty acid esters; propylene glycol fatty acid esters such as propylene glycol monostearate; polyoxyethylene hydrogenated castor oil 40 (HCO-40), polyoxyethylene hydrogenated castor oil 50 (HCO-50), polyoxyethylene hardened Hardened castor oil derivatives such as castor oil 60 (HCO-60), polyoxyethylene hydrogenated castor oil 80; polyoxyethylene (20) sorbitan (polysorbate 20) monolaurate, monostearate Polyoxyethylene sorbitan fatty acid esters such as polyoxyethylene (20) sorbitan (polysorbate 60), polyoxyethylene monooleate (20) sorbitan (polysorbate 80), polyoxyethylene (20) sorbitan isostearate; Glyceryl alkyl ether; alkyl glucoside; polyoxyalkylene alkyl ether such as polyoxyethylene cetyl ether; amines such as stearylamine and oleylamine; polyoxyethylene / methylpolysiloxane copolymer; And silicone surfactants such as lauryl PEG-9 polydimethylsiloxyethyl dimethicone and PEG-9 polydimethylsiloxyethyl dimethicone.
増粘剤としては、例えば、グアーガム、ローカストビーンガム、カラギーナン、キサンタンガム、カルボキシメチルセルロース、ヒドロキシメチルセルロース、ヒドロキシエチルセルロース、ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロース、ポリビニルアルコール、ポリビニルピロリドン、カルボキシビニルポリマー、アクリル酸メタクリル酸アルキル共重合体、ポリエチレングリコール、ベントナイト、(アクリル酸ヒドロキシエチル/アクリロイルジメチルタウリンNa)コポリマー、(アクリロイルジメチルタウリンアンモニウム/ビニルピロリドン)コポリマーなどが挙げられる。 Examples of the thickener include guar gum, locust bean gum, carrageenan, xanthan gum, carboxymethylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, alkyl methacrylate methacrylate. Copolymers, polyethylene glycol, bentonite, (hydroxyethyl acrylate / acryloyldimethyltaurine Na) copolymer, (acryloyldimethyltaurine ammonium / vinylpyrrolidone) copolymer, and the like.
保存剤としては、安息香酸、安息香酸ナトリウム、デヒドロ酢酸、デヒドロ酢酸ナトリウム、パラオキシ安息香酸イソブチル、パラオキシ安息香酸イソプロピル、パラオキシ安息香酸ブチル、パラオキシ安息香酸エチル、パラオキシ安息香酸プロピル、パラオキシ安息香酸ベンジル、パラオキシ安息香酸メチル、フェノキシエタノールなどが挙げられる。 Preservatives include benzoic acid, sodium benzoate, dehydroacetic acid, sodium dehydroacetate, isobutyl paraoxybenzoate, isopropyl paraoxybenzoate, butyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, benzyl paraoxybenzoate, paraoxybenzoate Examples include methyl benzoate and phenoxyethanol.
pH調整剤としては、無機酸(塩酸、硫酸など)、有機酸(乳酸、乳酸ナトリウム、クエン酸、クエン酸ナトリウム、コハク酸、コハク酸ナトリウムなど)、無機塩基(水酸化カリウム、水酸化ナトリウムなど)、有機塩基(トリエタノールアミン、ジイソプロパノールアミン、トリイソプロパノールアミンなど)などが挙げられる。
キレート剤としては、EDTA・2ナトリウム塩、EDTA・カルシウム・2ナトリウム塩などが挙げられる。
Examples of pH adjusters include inorganic acids (hydrochloric acid, sulfuric acid, etc.), organic acids (lactic acid, sodium lactate, citric acid, sodium citrate, succinic acid, sodium succinate, etc.), inorganic bases (potassium hydroxide, sodium hydroxide, etc.) ) And organic bases (such as triethanolamine, diisopropanolamine, and triisopropanolamine).
Examples of the chelating agent include EDTA / disodium salt, EDTA / calcium disodium salt, and the like.
安定化剤としては、ポリアクリル酸ナトリウム、ジブチルヒドロキシトルエン、ブチルヒドロキシアニソールなどが挙げられる。 Examples of the stabilizer include sodium polyacrylate, dibutylhydroxytoluene, butylhydroxyanisole and the like.
添加剤は1種を単独で又は2種以上を組み合わせて使用できる。 An additive can be used individually by 1 type or in combination of 2 or more types.
<その他の有効成分>
本発明の外用組成物は、本発明の効果を損なわない範囲で、その他の有効成分を含むことができる。有効成分の具体例としては、例えば、保湿成分、抗炎症成分、抗菌成分、ビタミン類、ペプチド又はその誘導体、アミノ酸又はその誘導体、細胞賦活化成分、老化防止成分、血行促進成分などが挙げられる。
<Other active ingredients>
The composition for external use of this invention can contain another active ingredient in the range which does not impair the effect of this invention. Specific examples of active ingredients include moisturizing ingredients, anti-inflammatory ingredients, antibacterial ingredients, vitamins, peptides or derivatives thereof, amino acids or derivatives thereof, cell activation ingredients, anti-aging ingredients, blood circulation promoting ingredients, and the like.
内服用形態 Internal form
本発明の有効成分である藻類または植物の抽出物は、薬学的に許容される担体と配合し、錠剤、カプセル剤、顆粒剤、散剤などの固形製剤;またはシロップ剤、注射剤などの液状製剤として経口または非経口的に投与することもできる。これらの製剤は、常法の添加剤を含んでいても良い。薬学的に許容される担体としては、製剤素材として慣用の各種有機あるいは無機担体物質が用いられ、固形製剤における賦形剤、滑沢剤、結合剤、崩壊剤;液状製剤における溶剤、溶解補助剤、懸濁化剤、等張化剤、緩衝剤、無痛化剤などとして配合される。また必要に応じて、防腐剤、抗酸化剤、着色剤、甘味剤などの製剤添加物を用いることもできる。 The algae or plant extract, which is an active ingredient of the present invention, is mixed with a pharmaceutically acceptable carrier, and is a solid preparation such as a tablet, capsule, granule or powder; or a liquid preparation such as a syrup or injection. Can also be administered orally or parenterally. These preparations may contain conventional additives. As the pharmaceutically acceptable carrier, various organic or inorganic carrier substances commonly used as pharmaceutical materials are used. Excipients, lubricants, binders, disintegrants in solid preparations; solvents, dissolution aids in liquid preparations , Suspending agents, isotonic agents, buffers, soothing agents and the like. Further, if necessary, preparation additives such as preservatives, antioxidants, colorants, sweeteners and the like can be used.
賦形剤の好適な例としては、例えばD−ソルビトール、マンニトール、キシリトールなどの糖アルコール、ブドウ糖、白糖、乳糖、果糖などの糖類、結晶セルロース、カルメロースナトリウム、クロスカルメロースナトリウム、リン酸水素カルシウム、コムギデンプン、コメデンプン、トウモロコシデンプン、バレイショデンプン、デキストリン、β−シクロデキストリン、軽質無水ケイ酸、酸化チタン、メタケイ酸アルミン酸マグネシウム、タルク、カオリンなどが挙げられる。 Preferable examples of the excipient include sugar alcohols such as D-sorbitol, mannitol and xylitol, sugars such as glucose, sucrose, lactose and fructose, crystalline cellulose, carmellose sodium, croscarmellose sodium, calcium hydrogen phosphate , Wheat starch, rice starch, corn starch, potato starch, dextrin, β-cyclodextrin, light silicic acid anhydride, titanium oxide, magnesium aluminate metasilicate, talc, kaolin and the like.
結合剤の好適な例としては、例えばメチルセルロース、エチルセルロース、ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロースなどのセルロース誘導体、ポリビニルピロリドン、ポリビニルアルコール、アクリル酸系高分子、ゼラチン、アラビアゴム、プルラン、アルファー化デンプン、カンテン、トラガント、アルギン酸ナトリウム、アルギン酸プロピレングリコールエステルなどが挙げられる。 Preferable examples of the binder include cellulose derivatives such as methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl pyrrolidone, polyvinyl alcohol, acrylic polymer, gelatin, gum arabic, pullulan, pregelatinized starch, agar , Tragacanth, sodium alginate, propylene glycol alginate, and the like.
崩壊剤の好適な例としては、例えばデンプン、低置換度ヒドロキシプロピルセルロース、カルボキシメチルセルロースカルシウム、クロスカルメロースナトリウム、ヒドロキシプロピルスターチ、部分アルファー化デンプンなどが挙げられる。溶剤の好適な例としては、例えば注射用水、アルコール、プロピレングリコール、マクロゴール、ゴマ油、トウモロコシ油などが挙げられる。 Preferable examples of the disintegrant include starch, low-substituted hydroxypropylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, hydroxypropyl starch, and partially pregelatinized starch. Preferable examples of the solvent include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil and the like.
滑沢剤の好適な例としては、ステアリン酸、ステアリン酸マグネシウム、ステアリン酸カルシウム、ステアリン酸ポリオキシル、セタノール、タルク、硬化油、ショ糖脂肪酸エステル、ジメチルポリシロキサン、ミツロウ、サラシミツロウなどが挙げられる。 Preferable examples of the lubricant include stearic acid, magnesium stearate, calcium stearate, polyoxyl stearate, cetanol, talc, hydrogenated oil, sucrose fatty acid ester, dimethylpolysiloxane, beeswax, and white beeswax.
溶解補助剤の好適な例としては、例えばポリエチレングリコール、プロピレングリコール、D−マンニトール、安息香酸ベンジル、エタノール、トリスアミノメタン、コレステロール、トリエタノールアミン、炭酸ナトリウム、クエン酸ナトリウムなどが挙げられる。懸濁化剤の好適な例としては、例えばステアリルトリエタノールアミン、ラウリル硫酸ナトリウム、ラウリルアミノプロピオン酸、レシチン、塩化ベンザルコニウム、塩化ベンゼトニウム、モノステアリン酸グリセリンなどの界面活性剤;例えばポリビニルアルコール、ポリビニルピロリドン、カルボキシメチルセルロースナトリウム、メチルセルロース、ヒドロキシメチルセルロース、ヒドロキシエチルセルロース、ヒドロキシプロピルセルロースなどの親水性高分子などが挙げられる。 Preferable examples of the solubilizer include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like. Suitable examples of the suspending agent include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate; for example, polyvinyl alcohol, Examples thereof include hydrophilic polymers such as polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose.
等張化剤の好適な例としては、例えば塩化ナトリウム、グリセリン、D−マンニトールなどが挙げられる。緩衝剤の好適な例としては、例えばリン酸塩、酢酸塩、炭酸塩、クエン酸塩などの緩衝液などが挙げられる。無痛化剤の好適な例としては、例えばベンジルアルコールなどが挙げられる。防腐剤の好適な例としては、例えばパラオキシ安息香酸エステル類、クロロブタノール、ベンジルアルコール、フェネチルアルコール、デヒドロ酢酸、ソルビン酸などが挙げられる。抗酸化剤の好適な例としては、例えば亜硫酸塩、アスコルビン酸などが挙げられる。 Preferable examples of the isotonic agent include sodium chloride, glycerin, D-mannitol and the like. Preferable examples of the buffer include buffer solutions such as phosphate, acetate, carbonate, citrate and the like. Preferable examples of the soothing agent include benzyl alcohol. Preferable examples of the preservative include paraoxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like. Preferable examples of the antioxidant include sulfite and ascorbic acid.
本発明の液状製剤または固形製剤を製造する方法としては、藻類または植物の抽出物を製剤原料と混合して液状製剤または固形製剤を製造する工程を含むものであれば特に限定はなく、公知の方法を使用することができる。 The method for producing the liquid preparation or solid preparation of the present invention is not particularly limited as long as it includes a step of producing a liquid preparation or solid preparation by mixing an algae or plant extract with a preparation raw material. The method can be used.
固形製剤調製例:
例えば、藻類または植物の抽出物と製剤原料を含む製剤組成物を練合し、スクリーンを通過させることで成型する押出造粒物を、粉砕し、整粒する方法、前記製剤組成物に練合水を加えバーチカルグラニュレーターによって成型する攪拌造粒の後に、コーミルを用いて粉砕・篩過する方法、及び前記製剤組成物をローラーコンパクターで圧縮した後、ロールグラニュレーターで粉砕し篩過する方法、撹拌造粒の後に、流動層乾燥する方法が例示される。
Example of solid preparation:
For example, a method of kneading a formulation composition containing an algae or plant extract and a formulation raw material, and pulverizing and sizing the extruded granulated product by passing through a screen, kneading the formulation composition After stirring and granulating by adding water and molding with a vertical granulator, pulverizing and sieving using a comil, and after compressing the formulation composition with a roller compactor, pulverizing and sieving with a roll granulator, A method of fluidized bed drying after stirring granulation is exemplified.
また、例えば、直打により製造する場合には、藻類または植物の抽出物と製剤原料を含む製剤組成物を混合した後、直接、打錠機に投入して打錠すればよい。 In addition, for example, in the case of producing by direct compression, after mixing an algae or plant extract and a pharmaceutical composition containing a pharmaceutical raw material, it may be directly put into a tableting machine for tableting.
食品 Food
本発明の藻類または植物の抽出物を含むLTBP-4産生促進剤は、食品または機能性食品に含有させて提供され得る。このような食品または機能性食品としては、米飯;そば、うどん、はるさめ、中華麺、即席麺、カップ麺を含む各種の麺類;清涼飲料、炭酸飲料、栄養飲料、果実飲料、乳酸飲料、スポーツ飲料等の飲料;カレールー、シチュー、各種スープ類;アイスクリーム、アイスシャーベット、かき氷等の冷菓;飴、クッキー、キャンディー、ガム、チョコレート、錠菓、スナック菓子、ビスケット、ゼリー、ジャム、クリーム、その他の焼き菓子等の菓子類;かまぼこ、はんぺん、ハム、ソーセージ等の水産・畜産加工食品;加工乳、発酵乳等の乳製品;サラダ油、てんぷら油、マーガリン、マヨネーズ、ショートニング、ホイップクリーム、ドレッシング等の油脂及び油脂加工食品;ソース、ドレッシング、味噌、醤油、たれ等の調味料;スープ、シチュー、サラダ、惣菜、ふりかけ、漬物;その他種々の形態の健康・栄養補助食品などが例示される。 The LTBP-4 production promoter containing the algal or plant extract of the present invention can be provided by being contained in a food or functional food. Examples of such foods or functional foods include cooked rice; various kinds of noodles including soba, udon, harusame, Chinese noodles, instant noodles, cup noodles; soft drinks, carbonated drinks, nutrition drinks, fruit drinks, lactic acid drinks, sports drinks Beverages such as Carrero, stew, various soups; Ice cream, ice sorbet, shaved ice and other frozen desserts; Strawberries, cookies, candy, gum, chocolate, tablet confectionery, snacks, biscuits, jellies, jams, creams, and other baked goods Sweets such as kamaboko, hampen, processed food such as ham, sausage; dairy products such as processed milk and fermented milk; fats and oils such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing Processed foods; seasonings such as sauces, dressings, miso, soy sauce, sauces; soups Stew, salad, delicatessen, sprinkle, pickles; health and nutrition supplement other various forms is illustrated.
さらには、本発明のLTBP-4産生促進剤を含有するサプリメント(散在、顆粒剤、ソフトカプセル、ハードカプセル、錠剤、チュアブル錠、速崩錠、シロップ、液剤等)を調製してもよい。 Furthermore, supplements (scattering, granules, soft capsules, hard capsules, tablets, chewable tablets, quick-disintegrating tablets, syrups, liquids, etc.) containing the LTBP-4 production promoter of the present invention may be prepared.
また、ペットなどの動物用の餌に対して本発明のLTBP-4産生促進剤を含有させることもできる。 Moreover, the LTBP-4 production promoter of this invention can also be contained with respect to animal foods, such as a pet.
食品には、必要に応じて、添加物が加えられる。このような添加物としては、例えばブドウ糖、果糖、ショ糖、マルトース、ソルビトール、ステビオサイド、ルブソサイド、コーンシロップ、乳糖、マンニット、デキストリン、クエン酸、クエン酸ソーダ、酒石酸、リンゴ酸、コハク酸、乳酸、L−アスコルビン酸、dl−α−トコフェロール、エリソルビン酸ナトリウム、グリセリン、プロピレングリコール、グリセリン脂肪酸エステル、ポリグリセリン脂肪酸エステル、ショ糖脂肪酸エステル、ソルビタン脂肪酸エステル、アラビアガム、カラギーナン、カゼイン、ゼラチン、ペクチン、寒天、ビタミンC、ビタミンB類、ビタミンE、ニコチン酸アミド、パントテン酸カルシウム、アミノ酸類、カルシウム塩類、界面活性剤、色素、香料、保存剤などが挙げられる。 Additives are added to the food as needed. Examples of such additives include glucose, fructose, sucrose, maltose, sorbitol, stevioside, rubusoside, corn syrup, lactose, mannitol, dextrin, citric acid, sodium citrate, tartaric acid, malic acid, succinic acid, lactic acid L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerol, propylene glycol, glycerol fatty acid ester, polyglycerol fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, gum arabic, carrageenan, casein, gelatin, pectin, Examples include agar, vitamin C, vitamin B, vitamin E, nicotinic acid amide, calcium pantothenate, amino acids, calcium salts, surfactants, pigments, fragrances, and preservatives.
本発明の藻類または植物抽出物を含むLTBP-4産生促進剤を用いて、LTBP-4をインビトロで作成すること、LTBP-4自体や弾性線維の研究への応用に用いることも可能である。本発明のLTBP-4産生促進剤を用いて、インビトロでLTBP-4を産生させ、人工弾性線維の培養用試薬、人工皮膚、皮膚モデル、人工血管などの人工組織の作製に用いることもできる。このようなインビトロでの研究や実施への応用に用いる場合、藻類または植物抽出物をそのままの形で用いることも好ましい。 It is also possible to prepare LTBP-4 in vitro using the LTBP-4 production promoter containing the algae or plant extract of the present invention, and to use it for the study of LTBP-4 itself and elastic fibers. The LTBP-4 production promoter of the present invention can be used to produce LTBP-4 in vitro and to produce artificial tissues such as artificial elastic fiber culture reagents, artificial skin, skin models, and artificial blood vessels. When used for such in vitro research and application, it is also preferable to use the algal or plant extract as it is.
LTBP-4をインビトロで産生させるには、藻類または植物抽出物を抽出したそのまま、あるいは乾燥重量で1%〜70%程度を含有する組成物を作成し、LTBP-4をインビトロで作成できる細胞と共存させる。このような組成物を、そのまま本発明のLTBP-4産生促進剤として用いることができる。LTBP-4を産生する具体的な方法としては、LTBP-4を産生することのできる細胞を、本発明のLTBP-4産生促進剤と接触させて培養することが含まれる。培養された細胞は、典型的には、細胞外にLTBP-4を分泌する。あるいは、インビボにおいて、LTBP-4の産生を促進させることもできる。用いられる細胞は、任意の動物に由来する細胞であり、好ましくは哺乳動物に由来するものである。このような細胞は、LTBP-4が存在する組織、例えば皮膚、血管、肺、子宮、心臓、腎臓、膵臓、精巣、卵巣、小腸、結腸、軟骨組織に由来する細胞、線維芽細胞、平滑筋細胞、上皮細胞、内皮細胞などの細胞が例示される。好ましくは、皮膚線維芽細胞又は血管平滑筋細胞が用いられる。インビトロにおけるLTBP-4産生には、生体由来の初代培養細胞、確立された細胞株、幹細胞などの未分化細胞の培養によって得られる細胞も用いることができる。 In order to produce LTBP-4 in vitro, a cell containing LTBP-4 can be prepared in vitro by preparing a composition containing about 1% to 70% by dry weight as extracted from an algal or plant extract. Coexist. Such a composition can be used as it is as an LTBP-4 production promoter of the present invention. A specific method for producing LTBP-4 includes culturing cells capable of producing LTBP-4 in contact with the LTBP-4 production promoter of the present invention. Cultured cells typically secrete LTBP-4 extracellularly. Alternatively, LTBP-4 production can be promoted in vivo. The cell to be used is a cell derived from any animal, preferably a mammal. Such cells are cells derived from tissues where LTBP-4 is present, such as skin, blood vessels, lungs, uterus, heart, kidney, pancreas, testis, ovary, small intestine, colon, cartilage, fibroblasts, smooth muscle Examples include cells such as cells, epithelial cells, and endothelial cells. Preferably, dermal fibroblasts or vascular smooth muscle cells are used. For in vitro LTBP-4 production, cells obtained by culturing undifferentiated cells such as primary cultured cells derived from living bodies, established cell lines, and stem cells can also be used.
LTBP-4を産生することのできる細胞としては、本来は、LTBP-4産生能を有さないか十分な量を発現することができない細胞であって、遺伝子導入によって、LTBP-4を産生するように操作された細胞も含まれる。LTBP-4遺伝子を発現し得るベクターを、細胞に導入することにより、LTBP-4産生能を獲得した細胞、あるいはLTBP-4産生能が改善された細胞が作製できる。 Cells that can produce LTBP-4 are originally cells that do not have LTBP-4 production ability or cannot express sufficient amounts, and produce LTBP-4 by gene transfer. Cells that have been manipulated in this way are also included. By introducing a vector capable of expressing the LTBP-4 gene into a cell, a cell having acquired LTBP-4 production ability or a cell having improved LTBP-4 production ability can be produced.
本発明の弾性線維形成促進剤は、さらに、DANCE(developmental arteries and neural crest epidermal growth factor(EGF)-like)発現増強因子(DANCE誘導因子)を含んでいてもよい。DANCE発現増強因子を用いることができ、例えば、ナンテンカズラエキス、シカクマメエキス、フィチン酸及びその生理学的に許容される塩、並びにL−ヒドロキシプロリン及びその生理学的に許容される塩等のDANCE発現増強因子;DANCE発現ベクター、またはDANCE誘導因子発現ベクターなどが挙げられる。 The elastic fiber formation promoter of the present invention may further contain a DANCE (developmental arteries and neural crest epidermal growth factor (EGF) -like) expression enhancing factor (DANCE inducing factor). DANCE expression enhancing factor can be used, for example, DANCE expression enhancement such as Nanten quail extract, winged bean extract, phytic acid and physiologically acceptable salts thereof, and L-hydroxyproline and physiologically acceptable salts thereof Factor: DANCE expression vector, DANCE inducing factor expression vector and the like.
次に、試験例、実施例等により本発明を具体的に説明するが、本発明は以下の実施例に限定されるものではない。 Next, the present invention will be specifically described with reference to test examples and examples, but the present invention is not limited to the following examples.
〔実施例1〜23、比較例1〜6〕(各種藻類および植物の抽出物)
表1〜3に示す市販の各種藻類および植物の抽出物を用いてそのままLTBP-4産生促進の評価試験に供した。
[Examples 1 to 23, Comparative Examples 1 to 6] (various algae and plant extracts)
It used for the evaluation test of LTBP-4 production promotion as it was using the extract of various commercially available algae and plants shown in Tables 1-3.
〔試験例1〕(LTBP-4産生促進の評価試験) [Test Example 1] (Evaluation test of LTBP-4 production promotion)
試験例1:
直接吸着法(ダイレクトELISA法)を利用して試験を実施した。
まず、96ウエルプレートにヒト線維芽細胞を1ウエルあたり3500個播種し、継代用培地(DMEM(Dulbecco’s Modified Eagle Medium、Invitrogenから購入)、10%ウシ胎仔血清(FBS、JRH Bioscienceから購入)、2mMのグルタミン、100units/mLのペニシリン、100g/mLのストレプトマイシン)にて37℃、5%CO2で48時間培養した。さらに、血清を含まない培地、すなわち、DMEM/F12、2mMのグルタミン、100units/mLのペニシリン、100g/mLのストレプトマイシンを用い、この無血清培地に、実施例1〜23および比較例1〜6で得た(表1〜3に記載)抽出物を0.001重量%加えて、あるいは加えないで、37℃、5%CO2で3日間培養した。培養した線維芽細胞の上清(または細胞溶解物)を96ウエルに添加し、4℃で一晩静置し固定相とした。その後、PBSを用いてウエルを洗浄し、ブロックエース(DSファーマバイオメディカルから購入)をPBSで4倍に希釈した溶液をウエルに添加しブロッキングを行った後、抗LTBP-4抗体溶液(サンタクルズから購入 SC-30360)を各ウエルに添加し、室温で1時間反応させた。さらに、PBSを用いてウエルを洗浄し、HRP標識された二次抗体溶液(Zymed Laboratories.Inc.,から購入 ZyMax81-1620)を添加して室温で1時間反応させ、その後発色基質を10分間反応させた。反応停止後の発色強度を吸光光度計(OD450nm)を用いて測定し、コントロールと比較して表わした数字を表1〜3に記載し、さらにグラフ化したものを図1〜3に示す。
Test Example 1:
The test was carried out using the direct adsorption method (direct ELISA method).
First, 3500 human fibroblasts were seeded per well in a 96-well plate, and subculture medium (DMEM (purchased from Dulbecco's Modified Eagle Medium, Invitrogen), 10% fetal bovine serum (purchased from FBS, JRH Bioscience) 2 mM glutamine, 100 units / mL penicillin, 100 g / mL streptomycin) at 37 ° C. and 5% CO 2 for 48 hours. Furthermore, a serum-free medium, that is, DMEM / F12, 2 mM glutamine, 100 units / mL penicillin, 100 g / mL streptomycin was used, and in this serum-free medium, Examples 1 to 23 and Comparative Examples 1 to 6 were used. The obtained extract (described in Tables 1 to 3) was cultured for 3 days at 37 ° C. and 5% CO 2 with or without 0.001% by weight of the extract. The cultured fibroblast supernatant (or cell lysate) was added to 96 wells and allowed to stand at 4 ° C. overnight to form a stationary phase. Subsequently, the wells were washed with PBS, blocked after adding Block Ace (purchased from DS Pharma Biomedical) 4 times with PBS to the wells, and then anti-LTBP-4 antibody solution (from Santa Cruz). Purchased SC-30360) was added to each well and allowed to react for 1 hour at room temperature. Further, the wells were washed with PBS, a secondary antibody solution labeled with HRP (ZyMax81-1620 purchased from Zymed Laboratories. Inc.) was added and allowed to react at room temperature for 1 hour, and then the chromogenic substrate was reacted for 10 minutes. I let you. The color intensity after the reaction was stopped was measured using an absorptiometer (OD450 nm), the numbers expressed in comparison with the control are shown in Tables 1 to 3, and further graphed are shown in FIGS.
〔組成物調製例1〕
化粧料組成物
以下の化粧料組成物の実施例中の数値の単位は「重量%」である。
[Composition Preparation Example 1]
Cosmetic composition The unit of the numerical value in the Example of the following cosmetic compositions is "weight%".
[組成物調製例2]
内服組成物
以下の食品組成物の処方例において、藻類または植物の抽出物の重量は乾燥重量である。
食品組成物の全処方において、藻類または植物の抽出物としては、エタノール抽出物が好ましい。
[Composition Preparation Example 2]
Oral Composition In the formulation examples of the following food compositions, the weight of the algae or plant extract is the dry weight.
In the whole formulation of the food composition, the algae or plant extract is preferably an ethanol extract.
この飲料は、1日3回の食事毎に1本摂取される。 This drink is taken once every three meals a day.
この錠剤は、1日3回の食事毎に3錠摂取される。 Three tablets of this tablet are taken every three meals a day.
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