JP2020019756A - Nitric oxide production promoter - Google Patents
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本発明は、特定の植物を原料として使用した一酸化窒素（ＮＯ）産生促進剤及びその利用に関するものである。より詳細には、ツバキ科ツバキ属に属するツバキの種子の脱脂粕の水性成分及び／又はアカショウマ等のチダケサシ属に属する植物の根茎の抽出物を有効成分として含有してなることを特徴とする血管内皮細胞でのＮＯ産生促進剤、及び、該剤を含有してなることを特徴とする血管内皮機能を改善するための経口組成物に関する。 The present invention relates to a nitric oxide (NO) production promoter using a specific plant as a raw material and its use. More specifically, a blood vessel characterized by containing, as an active ingredient, an aqueous component of defatted lees of camellia seeds belonging to the Camellia genus Camellia and / or an extract of a rhizome of a plant belonging to the genus Tadatake, such as red ginger. The present invention relates to an agent for promoting NO production in endothelial cells, and an oral composition comprising the agent, for improving vascular endothelial function.
血管内皮は血管の拡張及び／又は収縮、血管平滑筋細胞の増殖及び／又は抗増殖、血液凝固及び／又は抗凝固、血管の炎症及び／又は抗炎症、酸化及び／又は抗酸化等の様々な作用に影響を及ぼすことが知られている。そのため、血管内皮機能の低下は、高血圧、高脂血症、糖尿病、肥満等の原因となる。更に、血管内皮機能の低下は、運動不足、喫煙、塩分の過剰摂取等により引き起こされるため、日常生活における血管内皮機能の維持・改善が重要である。 The vascular endothelium may have a variety of effects, such as dilation and / or contraction of blood vessels, proliferation and / or anti-proliferation of vascular smooth muscle cells, blood coagulation and / or anticoagulation, inflammation and / or anti-inflammation of blood vessels, oxidation and / or antioxidation, etc. It is known to affect action. Therefore, a decrease in vascular endothelial function causes hypertension, hyperlipidemia, diabetes, obesity and the like. Furthermore, since the decrease in vascular endothelial function is caused by lack of exercise, smoking, excessive intake of salt, and the like, it is important to maintain and improve the vascular endothelial function in daily life.
ＮＯは、ＮＯ合成酵素（ＮＯＳ）によりアルギニンから産生される。ＮＯＳには、異なる３種のアイソフォームが存在し、誘導型一酸化窒素合成酵素（ｉＮＯＳ）、神経型一酸化窒素合成酵素（ｎＮＯＳ）及び血管内皮型一酸化窒素合成酵素（ｅＮＯＳ）が存在する（非特許文献１）。ｉＮＯＳは、カルシウム非依存性の酵素であり、炎症やストレスにより強く誘導され、主に免疫機能に関与している。ｉＮＯＳによる過剰なＮＯ産生は、敗血症や関節リウマチの原因となる。ｎＮＯＳは、カルシウム依存性の酵素であり、脳においてシナプス可塑性の調節に関与している。ｅＮＯＳは、カルシウム依存性の酵素であり、血管拡張に関与している。 NO is produced from arginine by NO synthase (NOS). There are three different isoforms of NOS, including inducible nitric oxide synthase (iNOS), neuronal nitric oxide synthase (nNOS), and vascular endothelial nitric oxide synthase (eNOS). (Non-Patent Document 1). iNOS is a calcium-independent enzyme, is strongly induced by inflammation and stress, and is mainly involved in immune functions. Excessive NO production by iNOS causes sepsis and rheumatoid arthritis. nNOS is a calcium-dependent enzyme and is involved in regulating synaptic plasticity in the brain. eNOS is a calcium-dependent enzyme and is involved in vasodilation.
血管内皮細胞でのＮＯ産生の低下は、骨格筋でのエネルギー消費を抑制し、肥満や糖尿病の進行を誘導することが報告されている（非特許文献２）。ＮＯによる血管拡張以外の作用は、血小板凝集抑制作用、血管壁への単球・マクロファージの結合の抑制、血小板の接着・凝集の阻害による血栓形成の抑制等がある。血管内皮細胞から放出される他の血管拡張因子として、プロスタグランジンＩ２、内皮由来血管過分極因子等があり、血管収縮因子として、エンドセリン、アンジオテンシンＩＩ、トロンボキサンＡ２等の様々な生理活性物質が産生・分泌されることが明らかとなっている。これらの因子のバランスにより血管機能を調節・維持している。 It has been reported that reduction of NO production in vascular endothelial cells suppresses energy consumption in skeletal muscle and induces progression of obesity and diabetes (Non-Patent Document 2). Actions other than vasodilation by NO include platelet aggregation inhibitory action, suppression of monocyte / macrophage binding to the blood vessel wall, suppression of thrombus formation by inhibition of platelet adhesion and aggregation, and the like. Other vasodilators released from vascular endothelial cells include prostaglandin I2, endothelium-derived vascular hyperpolarizing factor, and the like, and vasoconstrictors include various physiologically active substances such as endothelin, angiotensin II, and thromboxane A2. It is clear that it is produced and secreted. The balance of these factors regulates and maintains vascular function.
後述するツバキについては次のようなことが知られている。すなわち、ツバキは古くから観賞用園芸植物として利用されており、その種子から得られる油脂は燃料油、整髪料、高級食用油等に利用されてきた。又、木部は灰化し日本酒の醸造に利用され、実や種子の脱脂粕は農作物の肥料等に利用されてきた。実や種子の脱脂粕にはサポニンやタンニンが含まれ、この加工物を殺虫防虫剤（特許文献１）、農園芸用線虫防除剤（特許文献２）等に利用する提案がなされている。その他に、脂肪蓄積抑制作用（特許文献３）及び育毛作用（特許文献４）等が提案されている。しかしながら、ツバキの種子の脱脂粕に含まれる成分を経口的に摂取することにより、血管内皮細胞におけるＮＯ産生を促進する知見は見当たらない。 The following is known about camellia described later. That is, camellia has been used as an ornamental horticultural plant for a long time, and oils and fats obtained from its seeds have been used for fuel oils, hairdressings, high-grade edible oils, and the like. In addition, xylem is incinerated and used for sake brewing, and defatted cakes of fruits and seeds have been used as fertilizers for agricultural products. Saponin and tannin are contained in defatted meal of seeds and seeds, and proposals have been made to use this processed product as an insecticide and insecticide (Patent Document 1), a nematode control agent for agricultural and horticultural use (Patent Document 2), and the like. In addition, a fat accumulation inhibitory action (Patent Document 3) and a hair-growth action (Patent Document 4) have been proposed. However, there is no finding that orally ingesting a component contained in defatted lees of camellia seeds promotes NO production in vascular endothelial cells.
・・・（これ以降の段落番号を整理されたい。）
又、アカショウマ等のチダケサシ属に属する植物については、次のようなことが知られている。すなわち、チダケサシ属に属する植物はユキノシタ科に分類され、本発明に係るものは種々あるが、その代表例としてアカショウマ（学名：Ａｓｔｉｌｂｅ ｔｈｕｎｂｅｒｇｉｉ（ＳＩＥＢ．ｅｔ ＺＵＣＣ．）ＭＩＱ．）を挙げることができる。アカショウマは日本では本州、四国、九州各地の山地に自生する多年草で、その根茎を赤升麻とよび、升麻（キンポウゲ科のサラシナショウマ：Ｃｉｍｉｃｉｆｕｇａ ｓｉｍｐｌｅｘ ＷＯＲＭＳＫＪＯＲＤ等）の代用品として古来より下熱や解毒等の目的で利用されてきた。これまでに血中脂質改善剤（特許文献５）、血中コレステロール低減剤（特許文献６）、むくみ予防剤（特許文献７）、肥満抑制剤（特許文献８）等が提案されてきたが、アカショウマ等のチダケサシ属に属する植物の根茎に含まれる成分を経口的に摂取することにより、血管内皮細胞においてＮＯ産生を促進させる報告は見当たらない。
... (I would like to sort out the paragraph numbers after this.)
Further, the following is known about plants belonging to the genus Zinnia, such as red ginger. That is, plants belonging to the genus Zinnia are classified into the family Saxifragaceae, and there are various plants according to the present invention. A typical example thereof is red ginger (scientific name: Astilbe thunbergii (SIEB. Et ZUCC.) MIQ.). Red ginger is a perennial plant that grows naturally in mountains in Honshu, Shikoku, and Kyushu in Japan. It has been used for purposes such as detoxification. So far, blood lipid improving agents (Patent Document 5), blood cholesterol lowering agents (Patent Document 6), swelling preventive agents (Patent Document 7), obesity suppressants (Patent Document 8), and the like have been proposed. There is no report of promoting NO production in vascular endothelial cells by orally ingesting a component contained in the rhizome of a plant belonging to the genus Zinnia, such as red ginger.
かかる現状に鑑み、本発明は、血管内皮細胞でのＮＯ産生を促進するための、安全かつ安定な新規素材を開発するとともに、これを産業上有効活用できる態様の組成物を提供することを課題とした。 In view of the above situation, an object of the present invention is to develop a safe and stable new material for promoting NO production in vascular endothelial cells, and to provide a composition of an embodiment that can effectively utilize the material. And
前記課題を解決するために、本発明者らは、血管内皮細胞におけるＮＯ産生を促進するための素材について鋭意検討を重ねた結果、意外にもツバキの実や種子、又、アカショウマ等のチダケサシ属に属する植物の根が極めて有効であり、これらの植物にはヒトや動物のＮＯ産生を顕著に促進し得る成分が含まれており、これらの併用はより一層顕著に血管内皮細胞でのＮＯ産生を促進すること、更に、これを飲食品、医薬品、医薬部外品、飼料等の分野において有効利用できることを見出し、本発明を完成するに至った。 In order to solve the above problems, the present inventors have conducted intensive studies on materials for promoting NO production in vascular endothelial cells. As a result, surprisingly, camellia nuts and seeds, Are extremely effective, and these plants contain components that can significantly promote NO production in humans and animals, and the combination of these plants makes the production of NO in vascular endothelial cells even more remarkable. And found that it can be effectively used in the fields of foods and drinks, pharmaceuticals, quasi-drugs, feeds, and the like, and completed the present invention.
すなわち、本発明によれば、ツバキ科ツバキ属（Ｃａｍｅｌｌｉａ）に属するツバキ（Ｃａｍｅｌｌｉａ ｊａｐｏｎｉｃａ）の種子から採取される脱脂粕の水性成分を有効成分として含有してなることを特徴とするＮＯ産生促進剤が提供される。この水性成分は、前記脱脂粕を水及び／又は低級アルコールで抽出処理して得られる抽出物であることが望ましい。 That is, according to the present invention, a NO production promoter comprising, as an active ingredient, an aqueous component of defatted cake collected from the seeds of camellia (Camellia japonica) belonging to the Camellia genus Camellia. Is provided. The aqueous component is desirably an extract obtained by extracting the defatted cake with water and / or a lower alcohol.
前記水性成分にはサポニン類が含まれており、該サポニン類はとりわけカメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ａ１、カメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ａ２、カメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ｂ１、カメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ｂ２、カメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ｃ１及び／又はカメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ｃ２であることがより好ましい。 The aqueous component contains saponins, which are, inter alia, Camellia saponin A1, Camellia saponin A2, Camellia saponin B1, Camellia saponin (Camellia saponin, Camellia saponin B, Camellia saponin B). ) C1 and / or Camellia saponin C2 are more preferred.
又、アカショウマ等のチダケサシ属（Ａｓｔｉｌｂｅ）に属する植物あるいはその抽出物を有効成分として含有してなることを特徴とするＮＯ産生促進剤が提供される。この抽出物は、アカショウマ等のチダケサシ属（Ａｓｔｉｌｂｅ）に属する植物の根茎を水及び／又は低級アルコールで抽出処理して得られる抽出物であることが望ましい。 The present invention also provides a NO production promoter comprising a plant belonging to the genus Astilbe such as red ginger or an extract thereof as an active ingredient. This extract is desirably an extract obtained by extracting the rhizome of a plant belonging to the genus Astilbe such as red ginger with water and / or a lower alcohol.
前述のように処理して得られる抽出物は、量的な差はあるものの多種多様な成分を含有する複雑な組成物であり、ベルゲニン、アスチルビン、タキシフォリン、ポリフェノール類、タンニン類、各種水溶性成分（多糖、オリゴ糖、単糖、蛋白、ペプチド、アミノ酸、これらの複合体など）を含んでいる。 The extract obtained by processing as described above is a complex composition containing various components, albeit with a quantitative difference, such as bergenin, astilbin, taxifolin, polyphenols, tannins, and various water-soluble components. (Polysaccharides, oligosaccharides, monosaccharides, proteins, peptides, amino acids, and complexes thereof).
本発明の前記ＮＯ産生促進剤は、血管内皮細胞においてＮＯ産生を促進するものであることを特徴とする。 The NO production promoter of the present invention is characterized in that it promotes NO production in vascular endothelial cells.
本発明の他の特徴は、前記のＮＯ産生促進剤を含有してなる経口組成物が提供される点にある。この経口組成物の態様は飲食品であることが望ましい。 Another feature of the present invention is to provide an oral composition comprising the above-mentioned NO production promoter. This embodiment of the oral composition is desirably a food or drink.
又、本発明によれば、前記ＮＯ産生促進剤を含有してなることを特徴とする抗疲労のための経口組成物あるいは飲食品が提供される。 Further, according to the present invention, there is provided an oral composition for anti-fatigue, or a food or drink, comprising the NO production promoter.
本発明のさらなる他の特徴は、ツバキ科ツバキ属に属するツバキの種子の脱脂物を水及び／又は低級アルコールで処理した水性成分、及び／又は、アカショウマ等のチダケサシ属に属する植物の根茎を水及び／又は低級アルコールで処理した抽出物を有効成分として含有せしめるＮＯ産生促進剤の製造方法にある。 Still another feature of the present invention is that an aqueous component obtained by treating a defatted material of camellia seeds belonging to the Camellia genus Camellia with water and / or a lower alcohol, and / or a rhizome of a plant belonging to the genus Tadatake such as red ginger. And / or a method for producing a NO production promoter containing an extract treated with a lower alcohol as an active ingredient.
本発明のさらなる他の特徴は、血管内皮細胞でのＮＯ産生を促進するために前記ＮＯ産生促進剤あるいは経口組成物を経口投与又は摂取する方法にある。 Still another feature of the present invention is a method for orally administering or ingesting the NO production promoter or the oral composition for promoting NO production in vascular endothelial cells.
本発明に係るツバキの種子の脱脂粕から抽出された水性成分及び／又はアカショウマ等のチダケサシ属に属する植物の根茎の抽出物は、品質安定性に優れ、血管内皮細胞におけるＮＯ産生を促進することにより、レイノー症、動脈硬化症、バージャー病、高血圧症、脳血栓、脳梗塞、心筋梗塞、充血、鬱血、出血、貧血、認知症、免疫力の低下、脱毛症、倦怠感、疲労、筋肉痛、疲れ目、むくみ、肩こり、腰痛、皮膚のかゆみ、くま、くすみ、しもやけ、凍傷、シワ、肌の弾力低下、歯周病等の血管機能の障害により引き起される症状を予防及び／又は改善する効果、及び、発毛、育毛、養毛、滋養強壮、美肌等の効果を奏する。かかる効果は、前記加工物を有効成分として含有してなるＮＯ産生促進剤を経口的に摂取又は投与することによって顕著に発現される。したがって、本発明によれば、ツバキの種子の脱脂粕から抽出された水性成分及び／又はアカショウマ等のチダケサシ属に属する植物の根茎の抽出物を有効成分として含有してなる経口用組成物が提供され、これを血管内皮細胞でのＮＯ産生の低下により引き起こされる諸症状を予防及び／又は改善するための飲食品、医薬品、医薬部外品、飼料等として有効利用することができる。 The aqueous component extracted from the defatted cake of camellia seeds and / or the rhizome extract of a plant belonging to the genus Tadatake, such as red ginger, according to the present invention has excellent quality stability and promotes NO production in vascular endothelial cells. Due to Raynaud's disease, arteriosclerosis, Burger's disease, hypertension, cerebral thrombosis, cerebral infarction, myocardial infarction, hyperemia, congestion, bleeding, anemia, dementia, decreased immunity, alopecia, malaise, fatigue, muscle pain, Prevents and / or ameliorates symptoms caused by impaired vascular function such as tired eyes, swelling, stiff shoulders, back pain, itching of the skin, darkness, dullness, bruising, frostbite, wrinkles, reduced skin elasticity, periodontal disease and the like. It produces effects such as hair growth, hair growth, hair growth, nourishing tonic, and beautiful skin. Such effects are remarkably exhibited by orally ingesting or administering the NO production promoter containing the processed product as an active ingredient. Therefore, according to the present invention, there is provided an oral composition comprising, as an active ingredient, an aqueous component extracted from defatted cake of camellia seeds and / or an extract of a rhizome of a plant belonging to the genus Zinnia, such as red ginger. It can be effectively used as a food and drink, a pharmaceutical, a quasi-drug, a feed, etc. for preventing and / or improving various symptoms caused by a decrease in NO production in vascular endothelial cells.
以下に本発明を詳細に説明する。まず、本発明のＮＯ産生促進剤は、ヒトや動物の血管内皮細胞でのＮＯ産生を促進する機能を有するものであり、ツバキ科（Ｔｈｅａｃｅａｅ）のツバキ属（Ｃａｍｅｌｌｉａ）に属するツバキ（Ｃａｍｅｌｌｉａ ｊａｐｏｎｉｃａ）の種子から得られる脱脂粕の水性成分及び／又はアカショウマ等のチダケサシ属（Ａｓｔｉｌｂｅ）に属する植物の根茎の抽出物を有効成分として含有してなることを特徴とする。 Hereinafter, the present invention will be described in detail. First, the NO production promoter of the present invention has a function of promoting NO production in vascular endothelial cells of humans and animals. Camellia japonica belonging to the genus Camellia of theaceaceae family (Theaceae) is provided. And / or an extract of a rhizome of a plant belonging to the genus Astilbe such as red ginger and the like, as an active ingredient.
ツバキ属に属する植物として、一般に、ツバキ節に属するツバキ（Ｃａｍｅｌｌｉａ ｊａｐｏｎｉｃａ）等、チャ節に属するチャ（Ｃ．ｓｉｎｅｎｓｉｓ）等、サザンカ節に属するサザンカ（Ｃ．ｓａｓａｎｑｕａ）等、カワリバツバキ節に属するグランサムツバキ（Ｃ．ｇｒａｎｔｈａｍｉａｎａ）等、ヤナギバサザンカ節に属するヤナギバサザンカ（Ｃ．ｓａｌｉｃｉｆｏｌｉａ）等、ヒメサザンカ節に属するヒメサザンカ（Ｃ．ｌｕｔｃｈｕｅｎｓｉｓ）等が知られているが、本発明ではツバキ節に属するものを用いる。この例としてヤブツバキ（Ｃ．ｊａｐｏｎｉｃａ ｖａｒ．ｊａｐｏｎｉｃａ）、ユキツバキ（Ｃ．ｊａｐｏｎｉｃａ ｓｕｂｓｐ．ｒｕｓｔｉｃａｎａ）、リンゴツバキ（Ｃ．ｊａｐｏｎｉｃａ ｖａｒ．ｍａｃｒｏｃａｒｐａ）、ホウザンツバキ（Ｃ．ｊａｐｏｎｉｃａ ｓｕｂｓｐ．ｈｏｚａｎｅｎｓｉｓ）、ホンコンツバキ（Ｃ．ｈｏｎｇｋｏｎｇｅｎｅｓｉｓ）、トウツバキ（Ｃ．ｒｅｔｉｃｕｌａｔａ）、サルウィンツバキ（Ｃ．ｓａｌｕｅｎｅｎｓｉｓ）、ピタールツバキのピタルディー種（Ｃ．ｐｉｔａｒｄｉｉ ｖａｒ．ｐｉｔａｒｄｉｉ）及びユンナン種（Ｃ．ｐｉｔａｒｄｉｉ ｖａｒ．ｙｕｎｎａｎｉｃａ）、金花茶（Ｃ．ｎｉｔｉｄｉｓｓｉｍａ）、ヤマツバキ（ヤブツバキと同種）、山茶花（ヤブツバキと同種）、ヤクシマツバキ（リンゴツバキと同種）等を挙げることができる。これらのツバキは日本列島、中国山東半島、朝鮮半島等で自生し又は栽培されているものを適宜に利用すればよい。 As plants belonging to the Camellia genus, generally, Camellia japonica belonging to Camellia section, Cha (C. sinensis) belonging to Cha section, etc., Sasanqua (C. sasanqua) belonging to Sasanqua section, etc. (C. granthamiana) and the like, C. salicifolia and the like belonging to the genus Salixasanka, and C. lutchuensis and the like belonging to the genus Himesazanka are known. In the present invention, those belonging to the camellia sect. Used. Examples of this are camellia (C. japonica var. Japonica), snow camellia (C. japonica subsp. Rusticana), apple camellia (C. japonica var. Macrocarpa), and C. japonica (C. japonica). Hong Kongensis, C. reticulata, C. saluenensis, Citadel var. pitardii varieties of Pittal camellia and C. pitardii var. si. Japanese pine wasp (similar to Japanese camellia), sasanqua (similar to Japanese camellia), Kushimatsubaki can be mentioned (apple camellia and the like) and the like. These camellias may be appropriately grown or grown on the Japanese archipelago, the Shandong Peninsula of China, the Korean Peninsula, and the like.
本発明では、前記のツバキの種子を圧搾処理、ヘキサンやヘプタン等の疎水性有機溶媒又は液化二酸化炭素、液化プロパン等の液化ガスを用いた超臨界抽出処理等に供して、常法により油分を抽出した残渣である脱脂物を、前記水性成分を採取するための原料とすることが望ましい。ここで、ツバキの種子は早熟実及び成熟実のいずれから採取されるものでもよく、これらの種子を含む実全体を用いてもよいが、脱脂物及び有効成分の収量の点から成熟実又はその種子を用いることが望ましい。より好ましくは成熟ないしは完熟した種子を用いる。本発明では、成熟実から得られる種子を１〜２週間程度、天日等で乾燥させたものを用いるのがよい。 In the present invention, the above camellia seeds are subjected to squeezing treatment, subjected to a supercritical extraction treatment using a hydrophobic organic solvent such as hexane or heptane or liquefied carbon dioxide, or a liquefied gas such as liquefied propane, etc. It is desirable to use a defatted product as an extracted residue as a raw material for collecting the aqueous component. Here, camellia seeds may be collected from any of precocious fruit and mature fruit, and whole fruit including these seeds may be used. It is desirable to use seeds. More preferably, mature or ripe seeds are used. In the present invention, it is preferable to use a seed obtained from a matured fruit, which is dried for about 1 to 2 weeks under sunlight or the like.
前記脱脂粕の水性成分は任意の方法で製造することができるが、水及び／又は低級アルコールを用いて抽出処理するのが好ましい。低級アルコールは、その炭素数が大きくなると脱脂粕中の油性物質が抽出される傾向が大きくなるため、炭素数が５程度までのものが望ましく、メタノール、エタノール、ノルマルプロパノール、イソプロパノール、ノルマルブタノール、イソブタノール等を例示できる。炭素数が大きい低級アルコールを使用する場合は、脱脂粕中の油性成分の抽出を抑制するために含水率を高めるのがよい。例えば、ｎ−プロパノールの場合の含水率は約２０質量％〜約５０質量％とし、ｎ−ブタノールの場合の含水率は約４０質量％〜約７０質量％とする。望ましい抽出溶媒は水、メタノール及びエタノール、及び、これらの含水アルコール（含水率：０〜１００質量％）である。 The aqueous component of the defatted cake can be produced by any method, but is preferably subjected to an extraction treatment using water and / or a lower alcohol. As the lower alcohol has a larger carbon number, the oily substance in the defatted cake has a greater tendency to be extracted. Therefore, those having a carbon number of up to about 5 are desirable. Methanol, ethanol, normal propanol, isopropanol, normal butanol, isobutanol Butanol and the like can be exemplified. When a lower alcohol having a large number of carbon atoms is used, it is preferable to increase the water content in order to suppress the extraction of oily components in the defatted cake. For example, the water content for n-propanol is about 20% to about 50% by weight, and the water content for n-butanol is about 40% to about 70% by weight. Desirable extraction solvents are water, methanol and ethanol, and their hydrated alcohols (water content: 0 to 100% by mass).
脱脂粕を抽出するには、脱脂粕１質量部に対して前記抽出溶媒を約１質量倍〜約３０質量倍加え、常圧下又は１〜５気圧の加圧下、常温ないしは約１２０℃で、約１０分〜約３時間、必要に応じて撹拌して混合後、常温に冷却して濾過し、濾液を減圧乾燥、噴霧乾燥、凍結乾燥等の適当な手段により濃縮、乾燥する。尚、乾燥物は適宜に粉砕処理してもよい。このようにして本発明に係る脱脂粕の水性成分である淡黄色ないし黄赤色の固体を得ることができる。前記抽出方法は、一旦抽出処理した抽出残渣を繰り返し抽出処理したり、１〜３気圧の加圧下、約１００℃〜約１３０℃で行うことが望ましい。これにより本発明に係る水性成分の収量が増える。この水性成分はサポニン、タンニン等の多種成分を含む。 To extract the defatted cake, the extraction solvent is added in an amount of about 1 to about 30 times by weight based on 1 part by mass of the defatted cake, and at normal pressure or under a pressure of 1 to 5 atm. After stirring and mixing as necessary for 10 minutes to about 3 hours, the mixture is cooled to room temperature and filtered, and the filtrate is concentrated and dried by an appropriate means such as drying under reduced pressure, spray drying, freeze drying and the like. The dried product may be appropriately pulverized. Thus, a pale yellow to yellow-red solid, which is an aqueous component of the defatted lees according to the present invention, can be obtained. It is desirable that the extraction method is performed by repeatedly extracting the extraction residue once subjected to the extraction process, or at a temperature of about 100 ° C. to about 130 ° C. under a pressure of 1 to 3 atm. This increases the yield of the aqueous component according to the invention. This aqueous component contains various components such as saponin and tannin.
水性成分に含まれるサポニンとして、３β−［２−Ｏ−β−Ｄ−ガラクトピラノシル−３−Ｏ−（２−Ｏ−β−Ｄ−グルコピラノシル−α−Ｌ−アラビノピラノシル）−β−Ｄ−グルコピラヌロノシルオキシ］オレアナ−１２−エン−１６α，２２α，２８−トリオール２２−［（Ｚ）−２−メチル−２−ブテノアート］であるカメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ａ１、３β−［２−Ｏ−β−Ｄ−ガラクトピラノシル−３−Ｏ−（２−Ｏ−β−Ｄ−グルコピラノシル−α−Ｌ−アラビノピラノシル）−β−Ｄ−グルコピラヌロノシルオキシ）オレアナ−１２−エン−１６α，２２α，２８−トリオール２２−［（Ｅ）−２−メチル−２−ブテノアート］であるカメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ａ２、３β−［２−Ｏ−β−Ｄ−ガラクトピラノシル−３−Ｏ−（２−Ｏ−β−Ｄ−グルコピラノシル−α−Ｌ−アラビノピラノシル）−β−Ｄ−グルコピラヌロノシルオキシ）−１６α，２８−ジヒドロキシ−２２α−［［（Ｚ）−２−メチル−２−プテノイル］オキシ］オレアナ−１２−エン−２３−アールであるカメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ｂ１、３β−［［２−Ｏ−β−Ｄ−ガラクトピラノシル−３−Ｏ−（２−Ｏ−β−Ｄ−グルコピラノシル−α−Ｌ−アラビノピラノシル）−β−Ｄ−グルコピラヌロノシル）オキシ］−１６α，２８−ジヒドロキシ−２２α−［［（Ｅ）−２−メチル−２−ブテノイル］オキシ］オレアナ−１２−エン−２３−アールであるカメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ｂ２、３β−［２−Ｏ−β−Ｄ−ガラクトピラノシル−３−Ｏ−（２−Ｏ−β−Ｄ−グルコピラノシル−α−Ｌ−アラビノピラノシル）−β−Ｄ−グルコピラヌロノシルオキシ］オレアナ−１２−エン−１６α，２２α，２３，２８−テトラオール２２−［（Ｚ）−２−メチル−２−ブテノアート］であるカメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ｃ１、及び、３β−［２−Ｏ−β−Ｄ−ガラクトピラノシル−３−Ｏ−（２−Ｏ−β−Ｄ−グルコピラノシル−α−Ｌ−アラビノピラノシル）−β−Ｄ−グルコピラヌロノシルオキシ］オレアナ−１２−エン−１６α，２２α，２３，２８−テトラオール２２−［（Ｅ）−２−メチル−２−ブテノアート］であるカメリアサポニン（Ｃａｍｅｌｌｉａｓａｐｏｎｉｎ）Ｃ２等を例示することができ、これらのサポニンはツバキに特異的に含まれている。 As a saponin contained in the aqueous component, 3β- [2-O-β-D-galactopyranosyl-3-O- (2-O-β-D-glucopyranosyl-α-L-arabinopyranosyl)- β-D-Glucopyranuronosyloxy] oleana-12-ene-16α, 22α, 28-triol 22-[(Z) -2-methyl-2-butenoate] Camellia saponin A1,3β- [2-O-β-D-galactopyranosyl-3-O- (2-O-β-D-glucopyranosyl-α-L-arabinopyranosyl) -β-D-glucopyranuronosyloxy ) Camellia saponin A2,3β- [2) which is oleana-12-ene-16α, 22α, 28-triol 22-[(E) -2-methyl-2-butenoate] O-β-D-galactopyranosyl-3-O- (2-O-β-D-glucopyranosyl-α-L-arabinopyranosyl) -β-D-glucopyranuronosyloxy) -16α , 28-Dihydroxy-22α-[[(Z) -2-methyl-2-ptenoyl] oxy] oleana-12-en-23-al Camelliasaponin B1,3β-[[2-O-β -D-galactopyranosyl-3-O- (2-O-β-D-glucopyranosyl-α-L-arabinopyranosyl) -β-D-glucopyranuronosyl) oxy] -16α, 28 -Camelliasaponin B2,3β- [2, which is -dihydroxy-22α-[[(E) -2-methyl-2-butenoyl] oxy] oleana-12-en-23-al O-β-D-galactopyranosyl-3-O- (2-O-β-D-glucopyranosyl-α-L-arabinopyranosyl) -β-D-glucopyranuronosyloxy] oleana Camellia saponin C1, which is 12-ene-16α, 22α, 23,28-tetraol 22-[(Z) -2-methyl-2-butenoate], and 3β- [2-O-β-D -Galactopyranosyl-3-O- (2-O-β-D-glucopyranosyl-α-L-arabinopyranosyl) -β-D-glucopyranuronosyloxy] oleana-12-en-16α , 22α, 23,28-tetraol 22-[(E) -2-methyl-2-butenoate], such as Camellia saponin C2, and the like. Ponin is specifically contained in camellia.
一方、アカショウマ等のチダケサシ属に属する植物は、生の植物体あるいはその乾燥物を使用することができるが、その部位は根あるいは根茎が望ましく、根茎を含む根全体でもよく、その態様は根あるいは根茎に乾燥、細断あるいは粉砕等の加工処理を適宜に施したものが望ましい。 On the other hand, a plant belonging to the genus Zinnia, such as red ginger, can use a raw plant or a dried product thereof, but the site is desirably a root or rhizome, and the whole root including the rhizome may be used. It is desirable that the rhizome is appropriately subjected to processing such as drying, shredding, or pulverization.
チダケサシ属に属する植物として、アカショウマ（Ａｓｔｉｌｂｅ ｔｈｕｎｂｅｒｇｉｉ（ＳＩＥＢ．ｅｔ ＺＵＣＣ．）Ｍｉｑ．）、トリアシショウマ（Ａ．ｔｈｕｎｂｅｒｇｉｉ（ＳＩＥＢ．ｅｔ ＺＵＣＣ．）ＭＩＱ．ｖａｒ．ｃｏｎｇｅｓｔａ ＢＯＩＳＳ．（＝Ａ．ｏｄｏｎｔｏｐｈｙｌｌａ ＭＩＱ．））、アワモリショウマ（Ａ．ｊａｐｏｎｉｃａ（ＭＯＲＲ．ｅｔ ＤＥＣＮＥ．）Ａ．ＧＲＡＹ）、チダケサシ（Ａ．ｍｉｃｒｏｐｈｙｌｌａ ＫＮＯＬＬ）等を例示することができる。本発明においてはアカショウマ、トリアシショウマが好適であり、望ましくはアカショウマである。 As plants belonging to the genus Chidaceae, red ginger (Astilbe thunbergii (SIEB. Et ZUCC.) Miq.) And trillium (A. thunbergii (SIEB. Et ZUCC.) MIQ. Var. Congesta BOISS. )), A. japonica (MORR. Et DECNE.) A. GRAY), Chidake Sashimi (A. microphylla KNOLL) and the like. In the present invention, red ginseng and triascenta are preferable, and red ginseng is preferable.
前記抽出処理にあたって、抽出溶媒としては水、親水性有機溶剤又はこれらの混合溶剤を用いるのがよい。親水性有機溶剤としてメタノール、エタノール、プロパノール、イソプロパノール等の低級一価アルコール類、プロピレングリコール、１，３−ブタンジオール、グリセリン等の多価アルコール類、アセトン、メチルエチルケトン、エーテル、石油エーテル、酢酸エチル及びこれらの含水物や混合物を例示することができる。本発明の所望の効果を奏するための抽出物（エキス）を効率的に得るには、エタノール、アセトン、及びこれらの含水物を抽出用溶媒とすることが望ましい。なお、該含水物の水分含量は、例えば、エタノールの場合では１〜９９質量％、より好ましくは１０〜５０質量％であり、アセトンの場合には１〜５０質量％、より好ましくは１０〜３０質量％である。これらの範囲を外れると本発明の所望の効果が減少し又は抽出物（エキス）の収量が低下する。又、以下に述べる精製物を簡便かつ効率的に得るには、例えば、含水エタノール又は含水アセトンで抽出し、該抽出物（エキス）を採取するのがよい。 In the extraction treatment, it is preferable to use water, a hydrophilic organic solvent, or a mixed solvent thereof as the extraction solvent. As a hydrophilic organic solvent, methanol, ethanol, propanol, lower alcohols such as isopropanol, propylene glycol, 1,3-butanediol, polyhydric alcohols such as glycerin, acetone, methyl ethyl ketone, ether, petroleum ether, ethyl acetate and These hydrates and mixtures can be exemplified. In order to efficiently obtain an extract (extract) for achieving the desired effects of the present invention, it is desirable to use ethanol, acetone, and a hydrate thereof as a solvent for extraction. The water content of the hydrate is, for example, 1 to 99% by mass, more preferably 10 to 50% by mass in the case of ethanol, and 1 to 50% by mass, more preferably 10 to 30% by mass in the case of acetone. % By mass. Outside of these ranges, the desired effect of the present invention is reduced or the yield of the extract (extract) is reduced. In addition, in order to obtain the purified product described below simply and efficiently, it is preferable to extract the extract with hydrated ethanol or hydrated acetone, for example.
抽出処理は前記原料に対して１〜１００質量倍程度の抽出溶媒を加え、常圧もしくは加圧下、常温又は加熱状態で、適宜に攪拌して１０分〜数日間抽出処理する。ついで不溶物を濾過又は遠心分離して除き本発明に係るエキス液を得ることができ、さらに該エキス液から減圧蒸留、噴霧乾燥、凍結乾燥等の公知の手段で溶媒を除去することによって本発明に係る抽出物（エキス）を得ることができる。又、前記抽出物を常法により、吸着剤（シリカゲル、ケイ酸マグネシウム、イオン交換樹脂、活性アルミナ、セルロース、活性炭等）による分画、溶剤分別、これらの組み合わせ等の公知の精製処理に供することによって精製物を得ることができる。本発明ではいずれの処理物も使用することができるが、後述する用途の利便性や本発明の効果の点から前記の抽出物あるいはその精製物が望ましい。 In the extraction treatment, an extraction solvent of about 1 to 100 times the mass of the raw material is added, and the extraction treatment is carried out for 10 minutes to several days by stirring appropriately at normal pressure or under pressure, at normal temperature or in a heated state. Next, the extract solution according to the present invention can be obtained by filtering or centrifuging off the insoluble matter, and the solvent is removed from the extract solution by a known means such as distillation under reduced pressure, spray drying, freeze drying, etc. Can be obtained. In addition, the extract may be subjected to a known purification treatment such as fractionation with an adsorbent (silica gel, magnesium silicate, ion exchange resin, activated alumina, cellulose, activated carbon, etc.), solvent fractionation, or a combination thereof by a conventional method. Thus, a purified product can be obtained. In the present invention, any of the treated products can be used, but the above-mentioned extract or a purified product thereof is desirable from the viewpoint of the convenience of use described below and the effects of the present invention.
前述のように処理して得られる生成物は、量的な差はあるものの多種多様な成分を含有する複雑な組成物であり、ベルゲニン、アスチルビン、タキシフォリン、ポリフェノール類、タンニン類、各種水溶性成分（多糖、オリゴ糖、単糖、蛋白、ペプチド、アミノ酸、これらの複合体等）を含んでいる。 The product obtained by processing as described above is a complex composition containing various components, albeit with a quantitative difference, such as bergenin, astilbin, taxifolin, polyphenols, tannins, and various water-soluble components. (Polysaccharide, oligosaccharide, monosaccharide, protein, peptide, amino acid, complex thereof, etc.).
本発明のＮＯ産生促進剤において、ツバキ由来の前記水性成分とアカショウマ等のチダケサシ属の植物に由来する前記抽出物を併用すると、血管内皮細胞でのＮＯ産生促進作用がさらに顕著に発現する。この場合のＮＯ産生促進剤における、両原料の配合比率は、所望の効果を奏するための使用目的と用途、製造コスト等により適宜に変動させることができる。ツバキ由来の前記水性成分とチダケサシ属植物由来の前記抽出物との好適な配合比率は、概ね２０：１〜１：２０であり、望ましくは約１０：１〜約１：１０である。 In the NO production promoter of the present invention, when the aqueous component derived from camellia is used in combination with the extract derived from a plant of the genus Zinnia, such as red ginger, the effect of promoting NO production in vascular endothelial cells is more remarkably exhibited. In this case, the mixing ratio of the two raw materials in the NO production promoter can be appropriately varied depending on the purpose of use and application for achieving the desired effect, production cost, and the like. The preferred blending ratio of the aqueous component derived from camellia and the extract derived from the genus Zinnia is generally about 20: 1 to 1:20, preferably about 10: 1 to about 1:10.
本発明のＮＯ産生促進剤は、その有効成分としての前記水性成分及び／又は前記抽出物を固体状、ペースト状又は液体状の形態となし、これをそのままＮＯ産生促進剤としてよいが、必要に応じて本発明のＮＯ産生促進剤が利用される用途における公知の添加物を併用して、常法により含有せしめて組成物として調製することもできる。ここで、公知の添加物は経口摂取するために通常利用されるものが望ましく、例えば、賦形剤、結合剤、崩壊剤、滑沢剤、湿潤剤、流動化剤、保存剤、界面活性剤、安定剤、希釈剤、溶解剤、等張化剤、殺菌剤、防腐剤、矯味剤、矯臭剤、着色剤、香料等の添加物質を使用でき、又、ＮＯ産生促進効果を有する既知成分やその含有素材を併用してもよい。 In the NO production promoter of the present invention, the aqueous component and / or the extract as an active ingredient is formed into a solid, paste or liquid form, which may be used as it is as an NO production promoter. Accordingly, the composition can be prepared by using a known additive in combination with a known additive in which the NO production promoter of the present invention is used and incorporating the additive by a conventional method. Here, the known additives are preferably those usually used for oral ingestion, for example, excipients, binders, disintegrants, lubricants, wetting agents, fluidizing agents, preservatives, surfactants Additives such as stabilizers, diluents, solubilizers, isotonic agents, bactericides, preservatives, flavoring agents, flavoring agents, coloring agents, and flavors can be used. You may use the containing material together.
血管内皮細胞でのＮＯ産生促進作用が既知の成分や素材としては、前記のもののほか黒酢、グルタチオン、イチョウ葉エキス、ヘスペリジン、シトルリン、ショウガ、黒ショウガ、ヒハツ、ブドウ茎抽出物、ブドウ果皮抽出物、トランスレスベラトロール、リンゴ果皮抽出物、ルチン、黒大豆ポリフェノール、ココアポリフェノール、ピクノジェノール、フラバンジェノール、ケイヒ、高麗人参、卵白ペプチド、鶏や豚又は魚由来のコラーゲンペプチド等を例示できる。尚、本発明はこれらの例示によって何ら限定されるものではない。 Ingredients and materials known to promote NO production in vascular endothelial cells include, in addition to those described above, black vinegar, glutathione, ginkgo biloba extract, hesperidin, citrulline, ginger, black ginger, hihats, grape stalk extract, grape skin extract , Trans resveratrol, apple peel extract, rutin, black soybean polyphenol, cocoa polyphenol, pycnogenol, flavangenol, calyx, ginseng, egg white peptide, chicken, pig or fish-derived collagen peptide. Note that the present invention is not limited by these examples.
本発明においては、前述したＮＯ産生促進剤をそのままの形態で飲食品、医薬品、医薬部外品、飼料、その他産業分野の様々な製品として利用することができ、あるいは該各種製品の配合原料の一部として使用する態様でも利用できる。とりわけＮＯ産生を促進するための経口組成物となすことが好ましく、この経口組成物の最も好適な態様は飲食品である。この例を以下に述べるが、本発明はこれにより限定されるものではない。 In the present invention, the above-mentioned NO production promoter can be used as it is as a food and drink, a pharmaceutical, a quasi-drug, a feed, and various other products in the industrial field, or as a compounding raw material for the various products. It can also be used in a mode used as a part. In particular, it is preferable to form an oral composition for promoting NO production, and the most preferred embodiment of the oral composition is a food or drink. This example is described below, but the present invention is not limited thereby.
本発明のＮＯ産生促進剤を経口組成物とする場合の形態は、顆粒剤、錠剤、カプセル剤、液剤等の経口用製剤となすことが可能である。かかる製剤組成物における前記のツバキ由来水性成分やチダケサシ属植物由来抽出物の含有量は、併用原料の種類や含有量等により一律に規定し難いが、各々概ね０．００１質量％〜９０質量％程度、より望ましくは約０．０１質量％〜約７０質量％である。前記含有量が約０．００１質量％を下回ると本発明の所望効果が認められなくなり、約９０質量％を超えると実用的な製剤組成物を調製することが難しくなる。本発明のＮＯ産生促進剤は、これを望ましくは経口的に摂取又は投与する態様で使用する。経口摂取又は投与する場合の本発明のＮＯ産生促進剤の好適な量の目安は、該剤に含まれる前記水性成分ベースあるいは前記抽出物ベースで、ヒト成人１日あたり約１０ｍｇ〜約１，０００ｍｇ、望ましくは約３０ｍｇ〜約５００ｍｇ、更に望ましくは約５０ｍｇ〜約３００ｍｇである。 When the NO production promoter of the present invention is used as an oral composition, it can be made into oral preparations such as granules, tablets, capsules, and liquids. The content of the above-mentioned camellia-derived aqueous component and the extract of the genus Pteridophyta in such a pharmaceutical composition is difficult to uniformly define depending on the type and content of the combined raw material, but is generally about 0.001% by mass to 90% by mass. Degree, more desirably from about 0.01% to about 70% by weight. If the content is less than about 0.001% by mass, the desired effects of the present invention cannot be obtained. If the content exceeds about 90% by mass, it becomes difficult to prepare a practical pharmaceutical composition. The NO production promoter of the present invention is desirably used orally ingested or administered. A suitable amount of the NO production promoter of the present invention when taken orally or administered is, based on the aqueous component or the extract contained in the agent, from about 10 mg to about 1,000 mg per day for a human adult. , Preferably about 30 mg to about 500 mg, more preferably about 50 mg to about 300 mg.
本発明のＮＯ産生促進剤は、これ自体を飲食品、医薬品、飼料その他産業分野の様々な形態の製品とすることができ、あるいは、かかる製品の配合原料の一部とする態様でも利用することができる。実用的な用途としては飲食品がよい。 The NO production promoter of the present invention can be used as a product in various forms in the food and drink, pharmaceutical, feed, and other industrial fields, or can be used as a part of a compounding material of such a product. Can be. Food and drink are good for practical use.
飲食品の具体例として、野菜ジュース、果汁飲料、清涼飲料、茶等の飲料類、即席麺、スープ、ゼリー、プリン、ヨーグルト、ケーキプレミックス製品、菓子類、ふりかけ、味噌、醤油、ソース、ドレッシング、マヨネーズ、植物性クリーム、焼肉用たれや麺つゆ等の調味料、麺類、うどん、蕎麦、スパゲッティ、ハムやソーセージ等の畜肉魚肉加工食品、ハンバーグ、コロッケ、ふりかけ、佃煮、ジャム、牛乳、クリーム、バター、スプレッドやチーズ等の粉末状、固形状又は液状の乳製品、マーガリン、パン、ケーキ、クッキー、チョコレート、キャンディー、グミ、ガム、ゼリー等の各種一般加工食品のほか、粉末状、顆粒状、丸剤状、錠剤状、ソフトカプセル状、ハードカプセル状、ペースト状又は液体状の栄養補助食品、特定保健用食品、機能性食品、健康食品、濃厚流動食や嚥下障害用食品の治療食等を挙げることができる。 Specific examples of foods and beverages include beverages such as vegetable juices, fruit drinks, soft drinks, tea, instant noodles, soups, jellies, puddings, yogurts, cake premix products, confectionery, sprinkles, miso, soy sauce, sauces, and dressings , Mayonnaise, vegetable cream, seasonings such as sauces and noodle soup for yakiniku, noodles, udon, buckwheat, spaghetti, processed meat and meat products such as ham and sausage, hamburgers, croquettes, sprinkles, tsukudani, jam, milk, cream, Butter, powders such as spreads and cheeses, solid or liquid dairy products, margarine, bread, cake, cookies, chocolate, candy, gummy, gum, jelly, and other general processed foods, as well as powders, granules, Pill, tablet, soft capsule, hard capsule, paste or liquid dietary supplement, specific health Food, functional food, mention may be made of health food products, the concentrated liquid diet and dysphagia for food diet and the like.
尚、本発明の飲食品においては、これがツバキ科（Ｔｈｅａｃｅａｅ）のツバキ属（Ｃａｍｅｌｌｉａ）に属するツバキ（Ｃａｍｅｌｌｉａ ｊａｐｏｎｉｃａ）の種子の水性成分及び／又はアカショウマ等のチダケサシ属（Ａｓｔｉｌｂｅ）に属する植物の根茎の抽出物を有効成分として含有してなる旨、血管内皮細胞でのＮＯ産生を促進するためのものである旨のうち少なくとも１つの表示を付した態様とすることができる。 In the food and drink of the present invention, the aqueous component of the seeds of camellia (Camellia japonica) belonging to the camellia genus (Camellia) of theaceaceae (Theaceae) and / or the rhizome of a plant belonging to the genus Astilbe (Astilbe) such as red ginger. And at least one indication that the extract is contained as an active ingredient and that the extract is for promoting NO production in vascular endothelial cells.
これらの飲食品を製造するには、本発明のＮＯ産生促進剤と公知の原材料を用い、あるいは公知の原材料の一部を前記のＮＯ産生促進剤で置き換え、常法によって製造すればよい。例えば、本発明のＮＯ産生促進剤を、必要に応じてグルコース（ブドウ糖）、デキストリン、乳糖、澱粉又はその加工物、セルロース粉末等の賦形剤、ビタミン、ミネラル、動植物や魚介類の油脂、たん白（動植物や酵母由来の蛋白質、その加水分解物等を含む）、糖質、色素、香料、酸化防止剤、界面活性剤、その他の食用添加物、各種栄養機能成分を含む粉末やエキス類等の食用素材とともに混合して粉末、顆粒、ペレット、錠剤等の形状に加工したり、常法により前記例の一般加工食品に加工処理したり、これらを混合した液状物をゼラチン、アルギン酸ナトリウム、カルボキシメチルセルロース等の被覆剤で被覆してカプセルに成形したり、飲料（ドリンク類）の形態に加工して、栄養補助食品や健康食品として利用することは好適である。とりわけ錠剤、カプセル剤やドリンク剤が望ましい。 In order to produce these foods and drinks, the NO production promoter of the present invention and known raw materials may be used, or a part of the known raw materials may be replaced with the aforementioned NO production promoter, and the production may be carried out by a conventional method. For example, the NO production promoter of the present invention may be used, if necessary, with glucose (dextrose), dextrin, lactose, starch or a processed product thereof, excipients such as cellulose powder, vitamins, minerals, oils and fats of animals, plants and seafood, and protein. White (including proteins and hydrolysates derived from animals and plants or yeast, their hydrolysates, etc.), carbohydrates, pigments, fragrances, antioxidants, surfactants, other edible additives, powders and extracts containing various nutritive functional ingredients, etc. Powdered, granulated, pelletized, processed into the form of tablets, etc. by mixing with edible materials, processed into the general processed food of the above example by a conventional method, and a liquid material obtained by mixing these is gelatin, sodium alginate, carboxylate It is suitable to be coated with a coating agent such as methylcellulose and molded into capsules, or processed into the form of beverages (drinks) and used as dietary supplements or health foods A. Particularly, tablets, capsules and drinks are desirable.
かかる飲食品に配合する本発明のＮＯ産生促進剤の比率は、飲食品の形態、本発明のＮＯ産生促進剤中の前記水性成分（ツバキ科ツバキ属ツバキの種子の水性成分）及び／又は前記抽出物（アカショウマ等のチダケサシ属に属する植物の根茎の抽出物）の含量、他の配合原料の種類や成分や配合量等のちがいにより一律に規定しがたいが、飲食品中の前記水性成分及び／又は前記抽出物の含量が約０．０１質量％〜約９０質量％、より望ましくは約１質量％〜約５０質量％となるように、本発明のＮＯ産生促進剤をその他の飲食品製造用公知原料と適宜に組み合わせて処方を設計し、常法に従い目的とする飲食品を調製すればよい。本発明の飲食品は、ヒト成人の場合１日あたりの前記水性成分の摂取量の目安を約１０ｍｇ〜約１，０００ｍｇ、望ましくは約３０ｍｇ〜約５００ｍｇ、更に望ましくは約５０ｍｇ〜約３００ｍｇとして任意の方法、例えば、食事の摂取と同時又は前後に、経口摂取、経管投与等の方法で体内に取り込むことができる。 The ratio of the NO production promoter of the present invention to be added to the food or drink depends on the form of the food or drink, the aqueous component in the NO production promoter of the present invention (the aqueous component of the seeds of Camellia Camellia spp.) And / or Although the content of the extract (extract of the rhizome of a plant belonging to the genus Chidacea such as red ginger), the type of other compounding ingredients, the components, the amount of the compounding and the like are difficult to uniformly define, but the aqueous component in the food or drink is difficult. And / or other foods and beverages so that the content of the extract is about 0.01% to about 90% by weight, more preferably about 1% to about 50% by weight. The formulation may be designed by appropriately combining with known raw materials for production, and the intended food or drink may be prepared according to a conventional method. The food and drink of the present invention may have a guideline of the amount of the aqueous component per day for human adults of about 10 mg to about 1,000 mg, preferably about 30 mg to about 500 mg, and more preferably about 50 mg to about 300 mg. , For example, at the same time as or before and after ingestion of a meal, oral ingestion, tube administration and the like.
本発明のＮＯ産生促進剤を用いる医薬品は、前記のＮＯ産生促進剤に本発明の趣旨に反しない公知の賦形剤や添加物を適宜に加え、常法により加工して錠剤、カプセル剤、顆粒剤、散剤、液剤等の製剤となすことができる。経口あるいは経腸投与して、ＮＯ産生の促進あるいは前記諸症状の予防又は治療のために適用する。本発明のＮＯ産生促進剤の配合量はその形態や前記医薬製剤の種類、形態、用法及び用量等により一律に設定し難いが、前記水性成分及び／又は前記抽出物の含量として概ね０．００１質量％〜５０質量％である。経口投与する場合の摂取量はとくに限定されるものではないが、例えば、前記水性成分をベースにして、ヒト成人１日あたり約０．１ｍｇ〜約１，０００ｍｇ、望ましくは約１ｍｇ〜約５００ｍｇ、更に望ましくは約１０ｍｇ〜約３００ｍｇである。 Pharmaceutical products using the NO production promoter of the present invention are appropriately added with known excipients and additives not inconsistent with the gist of the present invention to the NO production promoter, processed by a conventional method, tablets, capsules, Preparations such as granules, powders, and liquids can be made. Oral or enteral administration is applied to promote NO production or prevent or treat the above-mentioned various symptoms. It is difficult to uniformly set the amount of the NO production promoter of the present invention depending on the form, type, form, usage, dosage, and the like of the pharmaceutical preparation, but the content of the aqueous component and / or the extract is generally 0.001. % By mass to 50% by mass. The oral intake is not particularly limited. For example, about 0.1 mg to about 1,000 mg, preferably about 1 mg to about 500 mg per day for a human adult, based on the aqueous component. More preferably, it is about 10 mg to about 300 mg.
又、本発明のＮＯ産生促進剤をペットフードや家畜用飼料に適用するには、前記飲食品の場合と同様に、公知の各種飼料や飲用水に配合したり、公知の原材料、添加物とともに錠剤状、顆粒状、カプセル状等の製剤形態のものに加工することができる。これらの場合、本発明のＮＯ産生促進剤の配合量及び摂取量は、前記水性成分の含量として約０．０１質量％〜約９０質量％、より望ましくは約１質量％〜約５０質量％であり、１日あたりの摂取量の目安は、前記水性成分を基準として、適用動物の体重（ｋｇ）あたり約０．１ｍｇ〜約１００ｍｇ、より望ましくは約０．５ｍｇ〜約５０ｍｇである。 Further, in order to apply the NO production promoter of the present invention to pet food or livestock feed, as in the case of the food or drink, it may be blended with various known feeds or drinking water, or together with known raw materials and additives. It can be processed into preparations such as tablets, granules and capsules. In these cases, the blending amount and intake amount of the NO production promoter of the present invention is about 0.01% to about 90% by weight, more preferably about 1% to about 50% by weight as the content of the aqueous component. The standard of daily intake is about 0.1 mg to about 100 mg, more preferably about 0.5 mg to about 50 mg, based on the weight of the aqueous component.
製造例１
長崎県五島産ヤブツバキ（Ｃ．ｊａｐｏｎｉｃａ ｖａｒ．ｊａｐｏｎｉｃａ）の乾燥種子を粗粉砕して蒸煮後、圧搾して圧搾油を分離した圧搾粕を得、次いで圧搾粕にノルマルヘキサンを加えて常法により抽出処理し、抽出液を分離して抽出粕を採取した。この抽出粕をノルマルヘキサンで洗浄して油分を取り除き脱脂物を採取した。この脱脂物１００ｇに水３００ｍＬを加え、常圧下、８５℃に加熱して１時間適宜に撹拌した後、室温まで冷却し、濾過して濾液を分離した。この濾過残渣に再度水２００ｍＬを加えて同様に加熱、攪拌、冷却後、濾過して濾液を採取した。両濾液を合わせて減圧下に濃縮し、凍結乾燥及び粉砕して、本発明に係る水性成分を含む粉末（試料Ｔ−１とする）１６．４ｇを得た。この粉末は、これを加水分解してＨＰＬＣ分析したところ、サポニンのアグリコンであるサポゲニンを１６．８％、フラボノールの一種であるケンフェロールを２．１％含むものであった。
Production Example 1
The dried seeds of the Japanese camellia (C. japonica var. Japonica) from Goto, Nagasaki are coarsely pulverized, steamed, and pressed to obtain a pressed cake from which pressed oil is separated. Then, normal hexane is added to the pressed cake and extracted by a conventional method. After the treatment, the extract was separated and the extracted cake was collected. This extracted cake was washed with normal hexane to remove oil, and a defatted product was collected. To 100 g of the defatted product, 300 mL of water was added, and the mixture was heated to 85 ° C. under normal pressure, stirred appropriately for 1 hour, cooled to room temperature, and filtered to separate a filtrate. To the filtration residue, 200 mL of water was added again, and the mixture was similarly heated, stirred, cooled, and then filtered to collect a filtrate. The two filtrates were combined, concentrated under reduced pressure, freeze-dried and pulverized to obtain 16.4 g of a powder containing an aqueous component according to the present invention (referred to as sample T-1). This powder was hydrolyzed and analyzed by HPLC. As a result, it was found that the powder contained 16.8% of sapogenin, an aglycone of saponin, and 2.1% of kaempferol, a kind of flavonol.
製造例２
屋久島産ヤクシマツバキ（Ｃ．ｊａｐｏｎｉｃａ ｖａｒ．ｍａｃｒｏｃａｒｐａ）の乾燥種子を製造例１に記載の方法で脱脂して脱脂物を採取した。この脱脂物１００ｇに水３００ｍＬを加え、２気圧の加圧下、１２０℃で２５分間加熱した後、室温まで冷却し、濾過して濾液を分離した。この濾過残渣に再度水２００ｍＬを加えて同様に加熱、攪拌、冷却後、濾過して濾液を採取した。両濾液を合わせて減圧下に濃縮し、凍結乾燥及び粉砕して、本発明に係る水性成分を含む粉末（試料Ｔ−２とする）１６．９ｇを得た。該粉末を製造例１と同様に加水分解してＨＰＬＣ分析した結果、サポゲニン含量は１５．１％であり、ケンフェロール含量は２．５％であった。
Production Example 2
Dry seeds of Yakushima-grown Japanese yam (C. japonica var. Macrocarpa) were defatted by the method described in Production Example 1 to collect defatted matter. To 100 g of the degreased product, 300 mL of water was added, and the mixture was heated at 120 ° C. for 25 minutes under a pressure of 2 atm, cooled to room temperature, and filtered to separate a filtrate. To the filtration residue, 200 mL of water was added again, and the mixture was similarly heated, stirred, cooled, and then filtered to collect a filtrate. The two filtrates were combined, concentrated under reduced pressure, freeze-dried and pulverized to obtain 16.9 g of a powder containing the aqueous component according to the present invention (referred to as sample T-2). The powder was hydrolyzed and analyzed by HPLC in the same manner as in Production Example 1. As a result, the sapogenin content was 15.1% and the kaempferol content was 2.5%.
製造例３
製造例１に記載の方法で得た脱脂物１００ｇに含水エタノール（含水率５０％）２５０ｍＬを加え、８０℃で１時間加熱還流した後、室温まで冷却し、濾過して濾液を分離した。この濾過残渣に再度含水エタノール（含水率５０％）２００ｍＬを加えて同様に加熱し、冷却後、濾過して濾液を採取した。両濾液を合わせて減圧下に濃縮し、凍結乾燥及び粉砕して、本発明に係る水性成分を含む粉末（試料Ｔ−３とする）１２．３ｇを得た。該粉末を製造例１と同様に加水分解してＨＰＬＣ分析した結果、サポゲニン含量は１２．５％であり、ケンフェロール含量は２．７％であった。
Production Example 3
To 100 g of the defatted product obtained by the method described in Production Example 1, 250 mL of aqueous ethanol (water content: 50%) was added, and the mixture was heated and refluxed at 80 ° C. for 1 hour, cooled to room temperature, filtered, and the filtrate was separated. 200 mL of aqueous ethanol (water content: 50%) was added again to the filtration residue, heated in the same manner, cooled, filtered, and the filtrate was collected. The two filtrates were combined, concentrated under reduced pressure, freeze-dried and pulverized to obtain 12.3 g of a powder containing an aqueous component according to the present invention (referred to as sample T-3). The powder was hydrolyzed and analyzed by HPLC in the same manner as in Production Example 1. As a result, the sapogenin content was 12.5% and the kaempferol content was 2.7%.
製造例４
製造例１に記載の方法で得た脱脂物１００ｇにエタノール（純度９９．５％）２００ｍＬを加え、８０℃で１時間加熱還流した後、室温まで冷却し、濾過して濾液を分離した。この濾過残渣に再度エタノール（純度９９．５％）２００ｍＬを加えて同様に加熱し、冷却後、濾過して濾液を採取した。両濾液を合わせて減圧下に濃縮し、凍結乾燥及び粉砕して、本発明に係る水性成分を含む粉末（試料Ｔ−４とする）４．３ｇを得た。該粉末を製造例１と同様に加水分解してＨＰＬＣ分析した結果、サポゲニン含量は１４．０％であり、ケンフェロール含量は２．５％であった。
Production Example 4
To 100 g of the defatted product obtained by the method described in Production Example 1, 200 mL of ethanol (purity: 99.5%) was added, and the mixture was heated under reflux at 80 ° C. for 1 hour, cooled to room temperature, filtered, and the filtrate was separated. 200 mL of ethanol (purity: 99.5%) was again added to the filtration residue, heated in the same manner, cooled, filtered, and the filtrate was collected. The two filtrates were combined, concentrated under reduced pressure, freeze-dried and pulverized to obtain 4.3 g of a powder containing an aqueous component according to the present invention (referred to as sample T-4). The powder was hydrolyzed in the same manner as in Production Example 1 and analyzed by HPLC. As a result, the sapogenin content was 14.0% and the kaempferol content was 2.5%.
製造例５
製造例１において、乾燥種子を未熟実（種子を含む実全体）におきかえること以外は同様に処理して、脱脂粕を得た後、これから水性成分を含む粉末（試料Ｔ−５とする）１２．８ｇを得た。該粉末を製造例１と同様に加水分解してＨＰＬＣ分析した結果、サポゲニン含量は１３．２％であり、ケンフェロール含量は２．４％であった。
Production Example 5
In Production Example 1, the same treatment was carried out except that the dried seeds were replaced with immature seeds (the whole seeds and seeds) to obtain defatted lees, and then powder containing an aqueous component (hereinafter referred to as sample T-5) 12 0.8 g was obtained. The powder was hydrolyzed and analyzed by HPLC in the same manner as in Production Example 1, and as a result, the sapogenin content was 13.2% and the kaempferol content was 2.4%.
製造例６
アカショウマ（Ａｓｔｉｌｂｅ ｔｈｕｎｂｅｒｇｉｉ（ＳＩＥＢ．ｅｔＺＵＣＣ．）ＭＩＱ．）の根茎を水洗し、長さ１〜２ｃｍ、幅３〜５ｍｍにみじん切りした後、天日干しで乾燥してアカショウマ乾燥物を調製した。このアカショウマ乾燥物１Ｋｇをステンレス製抽出釜に仕込み、含水率４５％の含水エタノール９Ｌを加え、適宜に撹拌しながら７０℃で６時間抽出処理した。この後、不溶の残渣を濾別してエキス液を採取し、該エキス液を減圧濃縮、凍結乾燥及び粉砕処理に供して赤褐色のエキス末（試料Ａ−１）を得た。
Production Example 6
The rhizome of red stinging ginger (Astilbe thunbergii (SIEB. EtZUCC.) MIQ.) Was washed with water, minced to a length of 1 to 2 cm and a width of 3 to 5 mm, and then dried by sun drying to prepare a dried red ginger. 1 kg of the dried red ginger was charged into a stainless steel extraction kettle, 9 L of aqueous ethanol having a water content of 45% was added, and the mixture was extracted at 70 ° C. for 6 hours with appropriate stirring. Thereafter, the insoluble residue was separated by filtration to collect the extract, and the extract was subjected to concentration under reduced pressure, freeze-drying and pulverization to obtain a red-brown extract powder (sample A-1).
製造例７
製造例１で調製したアカショウマ乾燥物１Ｋｇをステンレス製抽出釜に仕込み、水９Ｌを加え、適宜に撹拌しながら７０℃で６時間抽出処理した。この後、不溶の残渣を濾別してエキス液を採取し、該エキス液を減圧濃縮、凍結乾燥及び粉砕処理に供して赤褐色のエキス末（試料Ａ−２）を得た。
Production Example 7
1 Kg of the dried red ginger prepared in Production Example 1 was charged into a stainless steel extraction kettle, 9 L of water was added, and the mixture was extracted at 70 ° C. for 6 hours with appropriate stirring. Thereafter, the extract solution was collected by filtering off the insoluble residue, and the extract solution was concentrated under reduced pressure, freeze-dried and pulverized to obtain a red-brown extract powder (sample A-2).
製造例８
製造例１で調製したアカショウマ乾燥物１Ｋｇをステンレス製抽出釜に仕込み、エタノール９Ｌを加え、適宜に撹拌しながら７０℃で６時間抽出処理した。この後、不溶の残渣を濾別してエキス液を採取し、該エキス液を減圧濃縮、凍結乾燥及び粉砕処理に供して薄い赤褐色のエキス末（試料Ａ−３）を得た。
Production Example 8
1 kg of the dried red ginger prepared in Production Example 1 was charged into a stainless steel extraction kettle, 9 L of ethanol was added, and the mixture was extracted at 70 ° C. for 6 hours with appropriate stirring. Thereafter, the insoluble residue was separated by filtration to collect the extract, and the extract was subjected to concentration under reduced pressure, freeze-drying and pulverization to obtain a light red-brown extract powder (sample A-3).
試験例
前記製造例で得た各試料のＮＯ産生促進作用について以下に述べる方法で調べた。
Test Example The NO production promoting effect of each sample obtained in the above Production Example was examined by the method described below.
試験例１：ツバキ種子由来水性成分及びアカショウマ根茎由来抽出物によるウシ大動脈内皮細胞（ＢＡＥＣ）のＮＯ産生促進作用
動物細胞でのＮＯ産生促進作用を調べるため、ＢＡＥＣを使用した。ＢＡＥＣを９６Ｗｅｌｌプレートに１×１０４個／Ｗｅｌｌずつ播種し、３７℃で１日間培養した。その後、ＮＯ検出用蛍光プローブであるＤＡＦ−ＦＭ ＤＡを細胞に取り込ませ、試料Ｔ−１又は試料Ａ−１を１０、２５、５０、１００μｇ／ｍＬで添加し、３７℃で２４時間インキュベートした。又、これらを１０：１〜１：１０の割合で組み合わせた試料も１０μｇ／ｍＬで添加して同様に処理した。培養終了後、励起波長：４９０ｎｍ、蛍光波長：５２０ｎｍの蛍光強度を測定し、試料無添加の場合のＮＯ産生量を１としたときの相対値（活性）として、平均値±標準偏差（ｎ＝３）で表わした（表１）。
Test Example 1: Bovine aortic endothelial cell (BAEC) NO production promoting effect of camellia seed-derived aqueous component and red ginger rhizome-derived extract BAEC was used to examine the NO production promoting effect in animal cells. BAECs were seeded at 1 × 10 4 cells / well on a 96-well plate and cultured at 37 ° C. for 1 day. Thereafter, DAF-FMDA, which is a fluorescent probe for NO detection, was incorporated into the cells, and sample T-1 or sample A-1 was added at 10, 25, 50, or 100 μg / mL, and incubated at 37 ° C. for 24 hours. Also, a sample obtained by combining these at a ratio of 10: 1 to 1:10 was added at 10 μg / mL and treated similarly. After completion of the culture, the fluorescence intensity at an excitation wavelength of 490 nm and a fluorescence wavelength of 520 nm was measured, and the average value ± standard deviation (n = 3) (Table 1).
この結果を表１に示す。表１のデータから、本発明に係るツバキ種子由来水性成分及びアカショウマ根茎由来抽出物は濃度依存的にウシ大動脈内皮細胞のＮＯ産生促進作用を有することを確認した。更に、これらを１０：１〜１：１０の割合で混合した混合物も、各試料を単独で添加した場合と比較してより強いＮＯ産生促進作用を有することを確認した。 Table 1 shows the results. From the data in Table 1, it was confirmed that the aqueous component derived from camellia seed and the extract derived from rhizome rhizome according to the present invention have a concentration-dependent NO production promoting effect on bovine aortic endothelial cells. Furthermore, it was confirmed that a mixture obtained by mixing these at a ratio of 10: 1 to 1:10 also had a stronger NO production promoting action than the case where each sample was added alone.
試験例２：ツバキ（種子、実）由来水性成分及びアカショウマ根茎由来抽出物によるウシ大動脈内皮細胞（ＢＡＥＣ）のＮＯ産生促進作用
試験例１と同様にＢＡＥＣを培養した後、ＮＯ検出用蛍光プローブであるＤＡＦ−ＦＭ ＤＡを細胞に取り込ませた。試料Ｔ−１〜試料５又は試料Ａ−１〜試料Ａ−３を５０μｇ／ｍＬで添加し、３７℃で２４時間インキュベートした。培養終了後、励起波長：４９０ｎｍ、蛍光波長：５２０ｎｍの蛍光強度を測定し、試料無添加の場合のＮＯ産生量を１としたときの相対値（活性）として、平均値±標準偏差（ｎ＝３）で表わした（表２）。
Test Example 2: Promotion of NO production of bovine aortic endothelial cells (BAEC) by aqueous component derived from camellia (seed, fruit) and extract derived from rhizome rhizome After culturing BAEC in the same manner as in Test Example 1, using a fluorescent probe for NO detection Certain DAF-FM DAs were incorporated into cells. Sample T-1 to Sample 5 or Sample A-1 to Sample A-3 were added at 50 μg / mL and incubated at 37 ° C. for 24 hours. After completion of the culture, the fluorescence intensity at an excitation wavelength of 490 nm and a fluorescence wavelength of 520 nm was measured, and the average value ± standard deviation (n = 3) (Table 2).
この結果を表２に示す。表２のデータから、本発明に係るツバキの種子と実から得られる各脱脂物の水性成分はいずれもウシ大動脈内皮細胞の強いＮＯ産生促進作用を発現することを確認した。又、抽出溶媒が異なっても同様のＮＯ産生促進作用を有することを確認した。アカショウマ根茎由来抽出物によるＮＯ産生促進作用は、ツバキ由来水性成分と比べて小さかった。 Table 2 shows the results. From the data in Table 2, it was confirmed that all of the aqueous components of the defatted material obtained from camellia seeds and fruits according to the present invention exert a strong NO production promoting action on bovine aortic endothelial cells. Further, it was confirmed that the same NO production promoting action was obtained even when the extraction solvent was different. The NO production promoting effect of the extract from Rhizoma rhizome was smaller than that of the aqueous component derived from camellia.
試験例３：ツバキ種子由来水性成分及びアカショウマ根茎由来抽出物によるヒト臍帯静脈内皮細胞（ＨＵＶＥＣ）のＮＯ産生促進作用
ヒト細胞でのＮＯ産生促進作用を調べるため、ＨＵＶＥＣを用いた。ＨＵＶＥＣを９６Ｗｅｌｌプレートに１×１０４個／Ｗｅｌｌずつ播種し、３７℃で１日間培養した。その後、ＮＯ検出用蛍光プローブであるＤＡＦ−ＦＭ ＤＡを細胞に取り込ませ、試料Ｔ−１又は試料Ａ−１を２５、５０、１００μｇ／ｍＬで添加し、３７℃で２４時間インキュベートした。又、これらを１０：１〜１：１０の割合で混合した試料も２５μｇ／ｍＬで添加して同様処理した。培養終了後、励起波長：４９０ｎｍ、蛍光波長：５２０ｎｍの蛍光強度を測定し、試料無添加の場合のＮＯ産生量を１としたときの相対値（活性）として、平均値±標準偏差（ｎ＝３）で表わした（表３）。
Test Example 3: Promoting NO production of human umbilical vein endothelial cells (HUVEC) by an aqueous component derived from camellia seeds and extracts derived from rhizome rhizome HUVEC was used to examine the promotion of NO production by human cells. HUVECs were seeded at 1 × 10 4 cells / well on a 96-well plate and cultured at 37 ° C. for 1 day. Thereafter, DAF-FMDA, which is a fluorescent probe for NO detection, was incorporated into the cells, and Sample T-1 or Sample A-1 was added at 25, 50, or 100 μg / mL, and incubated at 37 ° C. for 24 hours. Also, a sample obtained by mixing these at a ratio of 10: 1 to 1:10 was added at 25 μg / mL and treated similarly. After completion of the culture, the fluorescence intensity at an excitation wavelength of 490 nm and a fluorescence wavelength of 520 nm was measured, and the average value ± standard deviation (n = 3) (Table 3).
この結果を表３に示す。表３のデータから、本発明に係るツバキ種子由来水性成分及びアカショウマ根茎由来抽出物はヒト臍帯静脈内皮細胞においても濃度依存的にＮＯ産生促進作用を有することを確認した。これらを１０：１〜１：１０の割合で混合した混合物も又、各単独を添加した場合と比較してより強いＮＯ産生促進作用を有することを確認した。 Table 3 shows the results. From the data in Table 3, it was confirmed that the aqueous component derived from camellia seed and the extract derived from rhizome rhizome according to the present invention also have a concentration-dependent NO production promoting effect on human umbilical vein endothelial cells. It was confirmed that a mixture obtained by mixing these at a ratio of 10: 1 to 1:10 also had a stronger NO production promoting action than the case where each was added alone.
試験例４：他の植物由来抽出物によるヒト臍帯静脈内皮細胞（ＨＵＶＥＣ）のＮＯ産生促進作用
本試験において、本発明のツバキ由来水性成分と比較するＮＯ産生促進物として、ヒハツエキスパウダー（Ｔｉｅ２ヒハツエキスパウダーＭＦ、丸善製薬社製）（比較試料１とする。）、ポリフェノール抽出物（リンゴ及びブドウ由来）（Ｖｉｎｉｔｒｏｘ Ｊ、ネキシラ社製）（比較試料２とする。）、ヘスペリジン（αＧヘスペリジンＰＡ−T、東洋精糖社製）（比較試料３とする。）及び大豆サポニン（サポニン，大豆由来、富士フィルム和光純薬社製）（比較試料４とする。）を用いた。大豆サポニンはサポゲニンとして８μｇ／ｍＬ（試料Ｔ−１ ５０μｇ／ｍＬと同量のサポゲニン量）を添加した。ＮＯ産生量の測定は試験例３と同様の方法を用いて行い、試料を添加しない場合のＮＯ産生量を１としたときの相対値（活性）として、平均値±標準偏差（ｎ＝３）で表わした（表４）。
Test Example 4: Effect of other plant-derived extracts on promotion of NO production of human umbilical vein endothelial cells (HUVEC) In this test, as a NO production promoter compared to the camellia-derived aqueous component of the present invention, Hihitsu extract powder (Tie2 Hihatsu extract powder) was used. MF, manufactured by Maruzen Pharmaceutical Co., Ltd.) (comparative sample 1), polyphenol extract (derived from apples and grapes) (Vinitrox J, manufactured by Nexira Co., Ltd.) (comparative sample 2), hesperidin (αG hesperidin PA-T, Toyo Seika Co., Ltd. (comparative sample 3) and soybean saponin (saponin, soybean-derived, Fujifilm Wako Pure Chemical Industries, Ltd.) (comparative sample 4) were used. As the soybean saponin, 8 μg / mL (the same amount of sapogenin as 50 μg / mL of sample T-1) was added as sapogenin. The measurement of the NO production amount was performed using the same method as in Test Example 3, and the relative value (activity) when the NO production amount when the sample was not added was set to 1, the average value ± standard deviation (n = 3) (Table 4).
この結果を表４に示す。表５のデータから、本発明に係るツバキ種子の脱脂物はヒト臍帯静脈内皮細胞において他のＮＯ産生促進剤よりも顕著なＮＯ産生促進作用を有することを確認した。 Table 4 shows the results. From the data in Table 5, it was confirmed that the degreased material of camellia seeds according to the present invention has a more remarkable NO production promoting effect on human umbilical vein endothelial cells than other NO production promoters.
試験例５：マウスにおけるＮＯ産生促進試験及び抗疲労試験
ＮＯ産生促進作用及び抗疲労作用を以下の方法で調べた。すなわち、６週齢ｄｄｙ系雄マウス（三協ラボサービス株式会社）を１週間予備飼育した後、１群４匹とし、蒸留水を投与する対照群、試料Ｔ−１投与群、試料Ａ−１投与群、及び、これらを１：１の割合で混合した試料投与群に群分けした。共通飼料（日本クレア株式会社製、商品名CE-2）を自由摂取させて、それぞれの試料２００ｍｇ／ｋｇ体重／日を毎日経口投与し、全てのマウスに１週間３回の遊泳運動を実施させて４週間飼育した。飼育２週目にマウスの尾に２ｇの錘をつけ、水槽で遊泳させ、疲労困憊してマウスの頭頂部が水面下に沈み込むような遊泳運動が継続不能になるまでの時間（限界遊泳時間）を測定した。また、飼育終了後にマウスの血液を採取し、遠心分離して得た血漿中のＮＯをＮＯ２／ＮＯ３ Ａｓｓａｙ Ｋｉｔ−Ｃ２（Ｃｏｌｏｒｉｍｅｔｒｉｃ）〜Ｇｒｉｅｓｓ Ｒｅａｇｅｎｔ Ｋｉｔ〜（同仁化学研究所製）を用いて測定した。血中ＮＯ濃度は対照群の値を１００としたときの相対値として、平均値±標準偏差で表わした（表６）。
Test Example 5: NO production promotion test and anti-fatigue test in mice The NO production promotion effect and anti-fatigue effect were examined by the following methods. That is, 6-week-old ddy strain male mice (Sankyo Lab Service Co., Ltd.) were preliminarily reared for 1 week, and each group was divided into 4 mice, and a control group to which distilled water was administered, a sample T-1 administration group, and a sample A-1. Each group was divided into an administration group and a sample administration group in which these were mixed at a ratio of 1: 1. A common feed (manufactured by Nippon Clea Co., Ltd., trade name CE-2) was freely ingested, and each sample was orally administered at a dose of 200 mg / kg body weight / day every day. All mice were allowed to perform swimming exercise three times a week. For 4 weeks. At the second week of breeding, a mouse was attached with a 2 g weight on its tail and allowed to swim in an aquarium. The time required for a swimming movement in which the head of the mouse became submerged under water and became unable to continue (limit swimming time) ) Was measured. After the rearing of the mice, the blood of the mice was collected, and the NO in the plasma obtained by centrifugation was measured using NO2 / NO3 Assay Kit-C2 (Colorimetric)-Grease Reagent Kit-(manufactured by Dojindo Laboratories). . The blood NO concentration was expressed as a mean value ± standard deviation as a relative value when the value of the control group was taken as 100 (Table 6).
この結果を表５に示す。表６のデータから、本発明に係るツバキ種子由来水性成分及びアカショウマ根茎由来抽出物は血中のＮＯ濃度を顕著に増加させること及び限界遊泳時間も著しく延長することを確認した。更に、これらを１：１の割合で混合した混合物の場合は、各単独での摂取と比較してより一層強い血中のＮＯ増強作用及び限界遊泳時間の延長作用を有することを確認した。 Table 5 shows the results. From the data of Table 6, it was confirmed that the aqueous component derived from camellia seeds and the extract derived from rhizome rhizome according to the present invention significantly increase the NO concentration in blood and significantly prolong the limit swimming time. Furthermore, it was confirmed that the mixture obtained by mixing them at a ratio of 1: 1 has a stronger effect of enhancing NO in blood and extending the limit swimming time as compared with the case of taking each alone.
試験例６：ヒトにおける抗疲労試験
試験に参加することに同意が得られた、日常的に運動を行う健常男性５０名（２１歳〜２８歳、平均年齢：２２．８歳）を、１群１０名で４群に分け、それぞれの群に各試料２００ｍｇを充填したゼラチンカプセルを１日１カプセル摂取してもらい、これを４週間続けた。被験者は、普段の生活通りに運動も継続的に実施し、疲労の評価として、Ｖｉｓｕａｌ Ａｎａｌｏｇ Ｓｃａｌｅ（ＶＡＳ）アンケートを実施した。なお、本試験は、プラセボ（マルトデキストリン）、前記の試料Ｔ−１、試料Ａ−１及び、これらを１：１の割合で混合した試料で行った。データは、平均値±標準偏差で表わした（表７）。
Test Example 6: A group of 50 healthy males (21 to 28 years old, average age: 22.8 years) who exercise daily and obtained consent to participate in the anti-fatigue test in humans Ten groups were divided into four groups, and each group received gelatin capsules filled with 200 mg of each sample, one capsule per day, which was continued for 4 weeks. The subjects also continuously exercised as usual, and performed a Visual Analog Scale (VAS) questionnaire as an evaluation of fatigue. This test was performed on a placebo (maltodextrin), the above-mentioned sample T-1, sample A-1, and a sample obtained by mixing these at a ratio of 1: 1. Data were expressed as mean ± standard deviation (Table 7).
この結果を表６に示す。表６のデータから、本発明に係るツバキ種子由来水性成分及びアカショウマ根茎由来抽出物は、疲労感を低減させることが明らかとなった。更に、これらを１：１の割合で混合した混合物は、混合前の以下の試料を単独摂取した場合と比較してより強い抗疲労効果を有することを確認した。 Table 6 shows the results. From the data in Table 6, it was clarified that the aqueous component derived from camellia seed and the extract derived from rhizome rhizome according to the present invention reduce the feeling of fatigue. Furthermore, it was confirmed that the mixture obtained by mixing them at a ratio of 1: 1 had a stronger anti-fatigue effect as compared with the case where the following sample before mixing was taken alone.
試作例１
本発明のＮＯ産生促進剤としての試料Ｔ−１〜５、試料Ａ−１〜３又は試料Ｔ−１／試料Ａ−１（１／１）混合物のいずれか１種をカプセル充填機に供して、常法により１粒あたり内容量が２００ｍｇのゼラチン被覆ハードカプセル製剤を試作した。その他の試料についても同様に処理して５種類のゼラチン被覆ハードカプセル製剤を試作した。これらのカプセル製剤は経口摂取が可能な栄養補助食品、医薬品等として使用することができる。
Prototype example 1
Any one of sample T-1 to 5, sample A-1 to 3, or sample T-1 / sample A-1 (1/1) mixture as the NO production promoter of the present invention is provided to a capsule filling machine. A gelatin-coated hard capsule preparation having a content of 200 mg per grain was produced by a conventional method. Other samples were treated in the same manner to prepare five kinds of gelatin-coated hard capsule preparations. These capsule preparations can be used as dietary supplements, medicines and the like that can be taken orally.
試作例２
本発明のＮＯ産生促進剤として試料Ｔ−１、試料Ａ−２又は試料Ｔ−１／試料Ａ−２（２／１）混合物：１５０部（質量基準。以下同様）、ミツロウ：４０部及び月見草油（英国エファモール社製）：８０部を約５０℃に加熱混合して均質にした後、カプセル充填機に供して、常法により１粒あたり内容量が２００ｍｇのゼラチン被覆ソフトカプセル製剤を試作した。これらのカプセル製剤は経口摂取可能な栄養補助食品として使用することができる。
Prototype example 2
As the NO production promoter of the present invention, sample T-1, sample A-2 or a mixture of sample T-1 / sample A-2 (2/1): 150 parts (by mass; hereinafter the same), beeswax: 40 parts and evening primrose Oil (manufactured by Ephamol Co., UK): 80 parts of the mixture were heated and mixed at about 50 ° C. to homogenize, and then supplied to a capsule filling machine to prepare a gelatin-coated soft capsule preparation having a content of 200 mg per grain by a conventional method. These capsule formulations can be used as dietary supplements that can be taken orally.
試作例３
本発明のＮＯ産生促進剤として試料Ｔ−３、試料Ａ−１又は試料Ｔ−３／試料Ａ−１（２／１）混合物：３０部、緑茶抽出物（ビーエイチエヌ（株）製）：０．５部、コーンスターチ（日本コーンスターチ（株）製）：１０５部、リン酸三カルシウム（米山化学工業（株）製）：５０部及びリボフラビン（ＤＳＭニュートリション・ジャパン（株）製）：７部を混合機に仕込み、１０分間攪拌混合した。この混合物を直打式打錠機に供して直径７ｍｍ、高さ４ｍｍ、質量１５０ｍｇ／個の素錠を作成し、ついでコーティング機でシェラック被膜を形成させて錠剤形状の食品を試作した。
Prototype example 3
As the NO production promoter of the present invention, sample T-3, sample A-1 or a mixture of sample T-3 / sample A-1 (2/1): 30 parts, green tea extract (manufactured by BNH Co., Ltd.): 0. 5 parts, corn starch (manufactured by Japan Corn Starch Co., Ltd.): 105 parts, tricalcium phosphate (manufactured by Yoneyama Chemical Industry Co., Ltd.): 50 parts, and riboflavin (manufactured by DSM Nutrition Japan KK): 7 parts And stirred and mixed for 10 minutes. This mixture was subjected to a direct compression tableting machine to prepare plain tablets having a diameter of 7 mm, a height of 4 mm, and a mass of 150 mg / piece, and then a shellac film was formed by a coating machine to produce a tablet-shaped food product.
試作例４
市販の栄養ドリンク１００ｍＬに本発明のＮＯ産生促進剤として試料Ｔ−３、試料Ａ−２又は試料Ｔ−１／試料Ａ−２（１／２）混合物：２００ｍｇ加えて十分に混合し飲料を試作した。これは冷蔵庫で１年間保存しても外観及び風味に異状及び違和感は認められなかった。尚、本品は、血管内皮細胞でのＮＯ産生促進、持久力の向上、疲労防止のために使用することができる。
Prototype example 4
200 mg of a sample T-3, sample A-2, or a mixture of sample T-1 / sample A-2 (1/2) as a NO production promoter of the present invention is added to 100 mL of a commercially available nutritional drink, and 200 mg of the mixture is thoroughly mixed to prepare a beverage. did. No abnormality or unpleasant appearance and flavor were observed in this product after storage in a refrigerator for one year. This product can be used for promoting NO production in vascular endothelial cells, improving endurance, and preventing fatigue.
試作例５
即席麺の製造工程において、公知の原料に本発明のＮＯ産生促進剤として試料Ｔ−１、試料Ａ−１又は試料Ｔ−２／試料Ａ−２（１／１）混合物を３００ｍｇ加えて即席麺を試作した。これは常温で６ヵ月間保存しても外観及び風味に異状及び違和感は認められなかった。尚、本品は、血管内皮細胞でのＮＯ産生促進、持久力の向上、疲労防止のために使用することができる。
Prototype example 5
In the instant noodle production process, 300 mg of the sample T-1, sample A-1 or sample T-2 / sample A-2 (1/1) mixture was added to the known raw material as the NO production promoter of the present invention, and the instant noodle was added. Was prototyped. No abnormal or uncomfortable appearance and flavor were observed even after storage at room temperature for 6 months. This product can be used for promoting NO production in vascular endothelial cells, improving endurance, and preventing fatigue.
本発明の、ツバキ科ツバキ属に属するツバキの種子の脱脂粕の水性成分及び／又はアカショウマ等のチダケサシ属に属する植物の根茎の抽出物を有効成分として含有してなるＮＯ産生促進剤は、これを経口摂取することにより血管内皮細胞でのＮＯ産生を促進する作用を有するため、ＮＯ産生の促進及び／又は血管内皮細胞でのＮＯ産生の低下に起因する症状を改善するための飲食品、医薬品、医薬部外品、飼料等に有効利用することができる。
The NO production promoter of the present invention comprising, as an active ingredient, an aqueous component of defatted lees of camellia seeds belonging to the Camellia genus Camellia and / or an extract of a rhizome of a plant belonging to the genus Tadatake, such as red ginger. Has an action of promoting NO production in vascular endothelial cells by orally ingesting it, so that foods and beverages and pharmaceuticals for promoting NO production and / or improving symptoms caused by a decrease in NO production in vascular endothelial cells , Quasi-drugs, feed, etc.
Claims (10)
A composition for improving endurance or a composition for preventing fatigue, comprising a NO production promoter obtained by the production method according to claim 9.
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| JP2003511094A (en) * | 1999-10-18 | 2003-03-25 | マッスルテク リサーチ アンド ディヴェロプメント インコーポレーション | Dietary supplements to increase lean body mass and physical fitness |
| JP2007131609A (en) * | 2005-11-11 | 2007-05-31 | Bhn Kk | Composition for reinforcing endurance, preventing or recovering fatigue, or strengthening nourishment |
| JP2008179592A (en) * | 2007-01-24 | 2008-08-07 | Ruiko Ito | Antioxidant, antiinflammatory agent and skin-lightening agent, and utilization thereof |
| JP2010265251A (en) * | 2009-05-13 | 2010-11-25 | Bhn Kk | Bloodstream-promoting/improving agent |
| JP2013184974A (en) * | 2012-03-06 | 2013-09-19 | Bhn Kk | Maillard reaction inhibitor and use thereof |
| CN105348375A (en) * | 2015-11-25 | 2016-02-24 | 华南理工大学 | Tea seed glucoprotein, preparation method therefor and application of tea seed glucoprotein |
| JP2016150910A (en) * | 2015-02-17 | 2016-08-22 | 日本メナード化粧品株式会社 | Endurance-improving agent |
| JP2017521482A (en) * | 2014-05-29 | 2017-08-03 | グゥアンヂョウ マグパイ ファーマシューティカルズ カンパニー リミテッド | Amantadine nitrate compound with neuroprotective effect and its preparation and medical application |
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| JP2003511094A (en) * | 1999-10-18 | 2003-03-25 | マッスルテク リサーチ アンド ディヴェロプメント インコーポレーション | Dietary supplements to increase lean body mass and physical fitness |
| JP2007131609A (en) * | 2005-11-11 | 2007-05-31 | Bhn Kk | Composition for reinforcing endurance, preventing or recovering fatigue, or strengthening nourishment |
| JP2008179592A (en) * | 2007-01-24 | 2008-08-07 | Ruiko Ito | Antioxidant, antiinflammatory agent and skin-lightening agent, and utilization thereof |
| JP2010265251A (en) * | 2009-05-13 | 2010-11-25 | Bhn Kk | Bloodstream-promoting/improving agent |
| JP2013184974A (en) * | 2012-03-06 | 2013-09-19 | Bhn Kk | Maillard reaction inhibitor and use thereof |
| JP2017521482A (en) * | 2014-05-29 | 2017-08-03 | グゥアンヂョウ マグパイ ファーマシューティカルズ カンパニー リミテッド | Amantadine nitrate compound with neuroprotective effect and its preparation and medical application |
| JP2016150910A (en) * | 2015-02-17 | 2016-08-22 | 日本メナード化粧品株式会社 | Endurance-improving agent |
| CN105348375A (en) * | 2015-11-25 | 2016-02-24 | 华南理工大学 | Tea seed glucoprotein, preparation method therefor and application of tea seed glucoprotein |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20220005162A (en) * | 2020-07-06 | 2022-01-13 | 황은경 | Pharmaceutical composition comprising the flower extract of camellia japonica as an effective component for prevention or treatment of thrombosis and health functional food comprising the same |
| KR102381447B1 (en) | 2020-07-06 | 2022-04-01 | 황은경 | Pharmaceutical composition comprising the flower extract of camellia japonica as an effective component for prevention or treatment of thrombosis and health functional food comprising the same |
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| JP7296611B2 (en) | 2023-06-23 |
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