JP2021183592A - Composition for improving vascular endothelial function - Google Patents
Composition for improving vascular endothelial function Download PDFInfo
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- JP2021183592A JP2021183592A JP2021066944A JP2021066944A JP2021183592A JP 2021183592 A JP2021183592 A JP 2021183592A JP 2021066944 A JP2021066944 A JP 2021066944A JP 2021066944 A JP2021066944 A JP 2021066944A JP 2021183592 A JP2021183592 A JP 2021183592A
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Abstract
Description
本発明は、プロシアニジンＢ１（ｐｒｏｃｙａｎｉｄｉｎ Ｂ１；以下、「ＰＢ１」とも言う）及びプロシアニジンＢ３（ｐｒｏｃｙａｎｉｄｉｎ Ｂ３；以下、「ＰＢ３」とも言う）を含有することを特徴する血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物に関し、前記プロシアニジンＢ１及びプロシアニジンＢ３が松樹皮由来であることを特徴とする血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物に関する。 The present invention is a composition for improving vascular endothelial function, vascular flexibility, which comprises procyanidin B1 (hereinafter also referred to as "PB1") and procyanidin B3 (hereinafter also referred to as "PB3"). Regarding the composition for improving sexuality and the composition for improving blood vessel-dependent vasodilatory function, the composition for improving vascular endothelial function, which is characterized in that procyanidin B1 and procyanidin B3 are derived from pine bark, for improving vascular flexibility. The present invention relates to a composition and a composition for improving a blood vessel-dependent vasodilator function.
近年、食生活やライフスタイルの変化に伴って生活習慣病が増加している。生活習慣病とは、高血圧症、高脂血症、糖尿病などの疾患に加え、狭心症、心筋梗塞、脳循環障害、悪性腫瘍など生活習慣を改善することによって発病を予防することができると考えられる疾患の総称である。したがって、生活習慣病に対しては、これらの症状を改善でき、長く安全に摂取できる食品による予防が重要であると考えられ、そのような効果を奏する食品の開発が期待されている。 In recent years, lifestyle-related diseases have been increasing with changes in eating habits and lifestyles. Lifestyle-related diseases can be prevented by improving lifestyle-related diseases such as angina, myocardial infarction, cerebral circulatory disorder, and malignant tumors, in addition to diseases such as hypertension, hyperlipidemia, and diabetes. It is a general term for possible diseases. Therefore, for lifestyle-related diseases, it is considered important to prevent these symptoms with foods that can be safely ingested for a long time, and the development of foods that have such effects is expected.
このような食品素材の一つとして、松樹皮抽出物がある。松樹皮抽出物は、プロアントシアニジンなどの抗酸化作用を有するポリフェノールを含有するとして、様々な食品に用いられてきた。また、松樹皮抽出物の研究により薬理作用を含む種々の作用が判明しており、それに基づいて種々の用途が提案されている。かかる用途としては、例えば皮膚改善剤（特許文献１）、腸内菌叢改善剤（特許文献２）、チロシナーゼ阻害剤（特許文献３）、ＤＮＡ保護剤（特許文献４）及び抗ストレス剤（特許文献５）が挙げられる。 One such food material is pine bark extract. Pine bark extract has been used in various foods as it contains polyphenols with antioxidant activity such as proanthocyanidins. In addition, research on pine bark extract has revealed various actions including pharmacological actions, and various uses have been proposed based on these actions. Such uses include, for example, a skin improving agent (Patent Document 1), an intestinal flora improving agent (Patent Document 2), a tyrosinase inhibitor (Patent Document 3), a DNA protective agent (Patent Document 4), and an anti-stress agent (Patent Document 4). Document 5) can be mentioned.
また、プロシアニジンを豊富に含む食品素材として、ココアやブドウが知られており、これらの食品素材には血管内皮機能改善作用があることが報告されている（特許文献６、７）。しかしながら、これらの食品素材に含まれるプロシアニジンが血管内皮に及ぼす影響については調べられていない。血管内皮機能改善には有効成分を毎日継続的に摂取することが必要であることから、継続的に摂取しやすく、少量の摂取であっても優れた効果を有する組成物の開発が求められている。 In addition, cocoa and grapes are known as food materials rich in procyanidins, and it has been reported that these food materials have an effect of improving vascular endothelial function (Patent Documents 6 and 7). However, the effects of procyanidins contained in these food materials on the vascular endothelium have not been investigated. Since it is necessary to take the active ingredient continuously every day to improve the vascular endothelial function, it is required to develop a composition that is easy to take continuously and has an excellent effect even if it is taken in a small amount. There is.
本発明は、優れた血管内皮機能改善効果を有する血管内皮機能改善用組成物を提供することを目的とする。 An object of the present invention is to provide a composition for improving vascular endothelial function having an excellent effect of improving vascular endothelial function.
本発明者らは、血管内皮機能改善効果について、鋭意調査・研究したところ、ＰＢ１及びＰＢ３を含有することにより、優れた血流依存性血管拡張機能改善効果が得られることを見出した。 The present inventors have diligently investigated and studied the effect of improving vascular endothelial function, and found that the inclusion of PB1 and PB3 provides an excellent effect of improving blood flow-dependent vasodilator function.
すなわち、本発明は、ＰＢ１及びＰＢ３を含有することを特徴とする組成物を摂取することで、優れた血管内皮機能改善効果、血管柔軟性向上効果、及び血流依存性血管拡張機能改善効果を発揮するという知見に基づいて完成したものである。 That is, the present invention has an excellent effect of improving vascular endothelial function, an effect of improving vascular flexibility, and an effect of improving blood flow-dependent vasodilator function by ingesting a composition characterized by containing PB1 and PB3. It was completed based on the knowledge that it will be exhibited.
本発明は要約すると以下の通りである。
［１］松樹皮由来のプロシアニジンＢ１及びプロシアニジンＢ３を含有することを特徴とする血流依存性血管拡張機能改善用組成物。
［２］松樹皮由来のプロシアニジンＢ１及びプロシアニジンＢ３を含有し、血流依存性血管拡張機能改善作用を有することを特徴とする血管内皮機能改善用組成物。
［３］松樹皮由来のプロシアニジンＢ１及びプロシアニジンＢ３を含有し、血流依存性血管拡張機能改善作用を有することを特徴とする血管柔軟性向上用組成物。
［４］錠状であることを特徴とする［１］〜［３］のいずれかに記載の組成物。
［５］プロシアニジンＢ１及びプロシアニジンＢ３を含有し、そのプロシアニジンＢ１とプロシアニジンＢ３の含有比がプロシアニジンＢ１に対してプロシアニジンＢ３が１．５以下であることを特徴とする血管内皮機能改善用組成物。
［６］プロシアニジンＢ１及びプロシアニジンＢ３を含有し、そのプロシアニジンＢ１とプロシアニジンＢ３の含有比がプロシアニジンＢ１に対してプロシアニジンＢ３が１．５以下であることを特徴とする血管柔軟性向上用組成物。
［７］プロシアニジンＢ１及びプロシアニジンＢ３を含有することを特徴とする血流依存性血管拡張機能改善用組成物。
［８］プロシアニジンＢ１とプロシアニジンＢ３の含有比がプロシアニジンＢ１に対してプロシアニジンＢ３が０．７６以下であることを特徴とする［５］〜［７］のいずれかに記載の組成物。
［９］プロシアニジンＢ１とプロシアニジンＢ３の含有比がプロシアニジンＢ１に対してプロシアニジンＢ３が０．６０以下であることを特徴とする［５］〜［７］のいずれかに記載の組成物。
［１０］プロシアニジンＢ１及びプロシアニジンＢ３が松樹皮由来であることを特徴とする［５］〜［９］のいずれかに記載の組成物。
［１１］錠状であることを特徴とする［５］〜［１０］のいずれかに記載の組成物。
The present invention is summarized as follows.
[1] A composition for improving a blood flow-dependent vasodilator function, which comprises procyanidin B1 and procyanidin B3 derived from pine bark.
[2] A composition for improving vascular endothelial function, which contains procyanidin B1 and procyanidin B3 derived from pine bark and has a blood flow-dependent vasodilatory function improving action.
[3] A composition for improving vascular flexibility, which contains procyanidin B1 and procyanidin B3 derived from pine bark and has a blood flow-dependent vasodilatory function improving action.
[4] The composition according to any one of [1] to [3], which is characterized by being tablet-shaped.
[5] A composition for improving vascular endothelial function, which contains procyanidin B1 and procyanidin B3, and the content ratio of procyanidin B1 to procyanidin B3 is 1.5 or less with respect to procyanidin B1.
[6] A composition for improving vascular flexibility, which contains procyanidin B1 and procyanidin B3, and the content ratio of procyanidin B1 to procyanidin B3 is 1.5 or less with respect to procyanidin B1.
[7] A composition for improving a blood flow-dependent vasodilator function, which comprises procyanidin B1 and procyanidin B3.
[8] The composition according to any one of [5] to [7], wherein the content ratio of procyanidin B1 to procyanidin B3 is 0.76 or less with respect to procyanidin B1.
[9] The composition according to any one of [5] to [7], wherein the content ratio of procyanidin B1 and procyanidin B3 is 0.60 or less with respect to procyanidin B1.
[10] The composition according to any one of [5] to [9], wherein procyanidin B1 and procyanidin B3 are derived from pine bark.
[11] The composition according to any one of [5] to [10], which is characterized by being tablet-shaped.
本発明によれば、ＰＢ１及びＰＢ３を含有することにより、血流依存性血管拡張機能改善効果に優れた組成物を提供することができる。また、本発明によれば、ＰＢ１及びＰＢ３を含有することにより、血流依存性血管拡張機能改善による血管柔軟性向上効果、血管内皮機能改善効果に優れた組成物を提供することができる。さらに、本発明によれば、松樹皮由来のＰＢ１及びＰＢ３を含有することにより、に優れた組成物を提供することができる血流依存性血管拡張機能改善効果、血流依存性血管拡張機能改善による優れた血管柔軟性向上効果、血管内皮機能改善効果に優れた組成物を提供することができる。 According to the present invention, by containing PB1 and PB3, it is possible to provide a composition having an excellent effect of improving blood flow-dependent vasodilator function. Further, according to the present invention, by containing PB1 and PB3, it is possible to provide a composition excellent in the effect of improving vascular flexibility and the effect of improving vascular endothelial function by improving the blood flow-dependent vasodilator function. Further, according to the present invention, the blood flow-dependent vasodilator function improving effect and the blood flow-dependent vasodilator function improving can provide an excellent composition by containing PB1 and PB3 derived from pine bark. It is possible to provide a composition having an excellent effect of improving vascular flexibility and an effect of improving vascular endothelial function.
本発明の血管内皮機能改善用組成物、血管柔軟性向上用組成物並びに血流依存性血管拡張機能改善用組成物（以下、これらの組成物をまとめて「本発明の組成物」とも言う）は、プロシアニジンＢ１（ＰＢ１）及びプロシアニジンＢ３（ＰＢ３）を含有することを特徴とする。本発明の組成物は、さらに必要に応じて、その他の成分を含有し得る。以下、各成分及び本発明の組成物について説明する。なお、以下に説明する構成は、本発明を限定するものでなく、本発明の主旨の範囲内で種々改変することができる。 The composition for improving vascular endothelial function, the composition for improving vascular flexibility, and the composition for improving blood flow-dependent vasodilatory function of the present invention (hereinafter, these compositions are collectively referred to as "the composition of the present invention"). Is characterized by containing procyanidin B1 (PB1) and procyanidin B3 (PB3). The composition of the present invention may further contain other components, if desired. Hereinafter, each component and the composition of the present invention will be described. The configuration described below is not limited to the present invention, and various modifications can be made within the scope of the gist of the present invention.
本発明の組成物は、プロシアニジンＢ１を含有する。プロシアニジンとは、フラボノイド類のフラバン−３−オールに属する化合物であり、エピカテキンあるいはカテキンが縮合したオリゴマーあるいはポリマー（２〜１５量体）である。プロシアニジンＢ１は、（−）エピカテキンと（＋）カテキンとがＣ４−Ｃ８結合した二量体である。本発明のプロシアニジンＢ１（ＰＢ１）は、合成物や松、リンゴ、ココアやブドウなどの植物由来物を用いることができ、使用時の安全性の観点から植物由来物が好ましく、ＰＢ１を豊富に含むことから、松がより好ましく、松樹皮を用いることが特に好ましい。 The composition of the present invention contains procyanidin B1. Procyanidins are compounds belonging to the flavonoids flavan-3-ols, and are epicatechins or oligomers or polymers (2 to 15-mer) in which catechins are condensed. Procyanidin B1 is a dimer in which (-) epicatechin and (+) catechin are bound to C4-C8. As the procyanidin B1 (PB1) of the present invention, synthetic products and plant-derived products such as pine, apple, cocoa and grape can be used, and plant-derived products are preferable from the viewpoint of safety during use, and PB1 is abundantly contained. Therefore, pine is more preferable, and pine bark is particularly preferable.
本発明においてＰＢ１の由来として使用できる松は、例えば、フランス海岸松、カラマツ、クロマツ、アカマツ、ヒメコマツ、ゴヨウマツ、チョウセンマツ、ハイマツ、リュウキュウマツ、ウツクシマツ、ダイオウマツ、シロマツ、カナダのケベック地方のアネダなどのマツ目に属する植物が挙げられるが、これらに限定されない。これらの中でも、食用として安全性が確認されており、ＰＢ１を高濃度で含有している観点から、南仏の大西洋沿岸などに生育している海洋性松であるフランス海岸松（Ｐｉｎｕｓ ｍａｒｉｔｉｍａ）が好ましい。 The pine that can be used as the origin of PB1 in the present invention includes, for example, French coastal pine, Karamatsu, Black pine, Japanese red pine, Himeko pine, Pinus parviflora, Japanese red pine, High pine, Ryukyu pine, Utsukushi pine, Daio pine, White pine, Aneda in the Quebec region of Canada, etc. Examples include, but are not limited to, plants belonging to the order Pinus parviflora. Among these, Pinus pinaster, which is a marine pine growing on the Atlantic coast of southern France, is preferable because it has been confirmed to be safe for food and contains a high concentration of PB1. ..
植物由来物の加工方法は特に限定されず、粉砕物、搾汁、抽出物等の処理物を使用することができる。粉砕物としては、粉末、顆粒等が挙げられる。搾汁や抽出物は、液状であってもよいが、ペースト状や乾燥粉末として用いることもできる。ペースト状や乾燥粉末とする場合は、そのもののみを用いて製造しても良いし、賦形剤と共に加工しても良い。本発明においては、血管内皮機能改善効果、血管柔軟性向上効果、及び血流依存性血管拡張機能改善効果の観点から、抽出により得ることが好ましい。以下、松樹皮抽出物の製造方法を例として植物由来物の抽出方法を説明する。 The processing method of the plant-derived product is not particularly limited, and a processed product such as a crushed product, a squeezed product, or an extract can be used. Examples of the crushed product include powder, granules and the like. The squeezed juice and the extract may be liquid, but can also be used as a paste or a dry powder. In the case of a paste or a dry powder, it may be produced by itself or processed together with an excipient. In the present invention, it is preferable to obtain it by extraction from the viewpoints of the effect of improving vascular endothelial function, the effect of improving vascular flexibility, and the effect of improving blood flow-dependent vasodilator function. Hereinafter, a method for extracting a plant-derived product will be described using a method for producing a pine bark extract as an example.
松樹皮抽出物は、松樹皮を溶媒で抽出して得られる。溶媒としては、例えば、水、有機溶媒、含水有機溶媒（含水エタノールなどの含水アルコール）が挙げられるが、これらに限定されない。抽出に用いる有機溶媒としては、通常天然物成分を抽出するのに際して許容される有機溶媒が用いられ、例えば、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、ブタン、アセトン、ヘキサン、シクロヘキサン、プロピレングリコール、含水エタノール、含水プロピレングリコール、エチルメチルケトン、グリセリン、酢酸メチル、酢酸エチル、ジエチルエーテル、ジクロロメタン、食用油脂、１，１，１，２−テトラフルオロエタン、１，１，２−トリクロロエテンなどが挙げられる。これらの溶媒は１種を単独で、又は２種以上を組合せて用いられ得る。抽出の際の温度は、室温から抽出溶媒の沸点以下の温度まで、適宜調整することができる。本発明においては、ＰＢ１を効率的に抽出する観点から、熱水、含水エタノール及び含水プロピレングリコールが好ましく用いられる。 The pine bark extract is obtained by extracting the pine bark with a solvent. Examples of the solvent include, but are not limited to, water, organic solvents, and hydrous organic solvents (hydrous alcohols such as hydrous ethanol). As the organic solvent used for extraction, an organic solvent usually acceptable for extracting natural product components is used, and for example, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, butane, etc. Acetone, hexane, cyclohexane, propylene glycol, hydrous ethanol, hydrous propylene glycol, ethylmethylketone, glycerin, methyl acetate, ethyl acetate, diethyl ether, dichloromethane, edible fats and oils, 1,1,1,2-tetrafluoroethane, 1, 1,2-Trichloroethane and the like can be mentioned. These solvents may be used alone or in combination of two or more. The temperature at the time of extraction can be appropriately adjusted from room temperature to a temperature below the boiling point of the extraction solvent. In the present invention, hot water, hydrous ethanol and hydrous propylene glycol are preferably used from the viewpoint of efficiently extracting PB1.
抽出方法は、通常天然物成分を抽出するのに際して許容される方法であれば特に限定されないが、例えば、加温抽出法、超臨界流体抽出法などの固液抽出法が挙げられる。 The extraction method is not particularly limited as long as it is usually an acceptable method for extracting natural product components, and examples thereof include solid-liquid extraction methods such as a heated extraction method and a supercritical fluid extraction method.
加温抽出法は、例えば、被験物質と溶媒とを接触させ、溶媒の沸点以下の温度などで処理して、被験物質に含まれる成分を溶媒に抽出する方法である。還流抽出法であってもよい。 The heated extraction method is, for example, a method in which a test substance and a solvent are brought into contact with each other and treated at a temperature equal to or lower than the boiling point of the solvent to extract components contained in the test substance into the solvent. It may be a reflux extraction method.
超臨界流体抽出法は、例えば、物質の気液の臨界点（臨界温度、臨界圧力）を超えた状態の流体である超臨界流体を用いて抽出を行う方法である。超臨界流体としては、二酸化炭素、エチレン、プロパン、亜酸化窒素（笑気ガス）などが挙げられるが、好ましくは二酸化炭素である。 The supercritical fluid extraction method is, for example, a method of extracting using a supercritical fluid which is a fluid in a state where the critical point (critical temperature, critical pressure) of the gas or liquid of a substance is exceeded. Examples of the supercritical fluid include carbon dioxide, ethylene, propane, nitrous oxide (laughing gas) and the like, and carbon dioxide is preferable.
超臨界流体抽出法では、目的成分を超臨界流体によって抽出する抽出工程と、目的成分と超臨界流体を分離する分離工程とを行う。分離工程では、圧力変化による抽出分離、温度変化による抽出分離、吸着剤・吸収剤を用いた抽出分離のいずれを行ってもよい。 In the supercritical fluid extraction method, an extraction step of extracting the target component with the supercritical fluid and a separation step of separating the target component and the supercritical fluid are performed. In the separation step, any of extraction separation by pressure change, extraction separation by temperature change, and extraction separation using an adsorbent / absorbent may be performed.
エントレーナー添加法による超臨界流体抽出を行ってもよい。この方法は、抽出流体に、例えば、エタノール、プロパノール、ｎ−ヘキサン、アセトン、トルエン、その他の脂肪族低級アルコール類、脂肪族炭化水素類、芳香族炭化水素類、ケトン類を２〜２０Ｗ／Ｖ％程度添加し、この流体を用いて超臨界流体抽出を行うことによって、ＯＰＣ（ｏｌｉｇｏｍｅｒｉｃ ｐｒｏａｎｔｈｏｃｙａｎｉｄｉｎ：オリゴメリック・プロアントシアニジン）、カテキン類などの目的とする抽出物の抽出溶媒に対する溶解度を飛躍的に上昇させる、又は分離の選択性を増強させる方法であり、効率的な松樹皮抽出物を得る方法である。 Supercritical fluid extraction by the entrainer addition method may be performed. In this method, for example, ethanol, propanol, n-hexane, acetone, toluene, other aliphatic lower alcohols, aliphatic hydrocarbons, aromatic hydrocarbons, and ketones are added to the extraction fluid at 2 to 20 W / V. By adding about% and performing supercritical fluid extraction using this fluid, the solubility of the target extract such as OPC (oligomeric fluorocyanidin) and catechins in the extraction solvent is dramatically increased. It is a method of increasing the selectivity of the pine bark or the separation, and is a method of obtaining an efficient pine bark extract.
超臨界流体抽出法は、比較的低い温度で操作できるため、高温で変質・分解する物質にも適用できるという利点、抽出流体が残留しないという利点、溶媒の循環利用が可能であるため、脱溶媒工程などが省略でき、工程がシンプルになるという利点がある。 Since the supercritical fluid extraction method can be operated at a relatively low temperature, it has the advantage that it can be applied to substances that deteriorate and decompose at high temperatures, the advantage that the extraction fluid does not remain, and the solvent can be recycled, so that the solvent is removed. There is an advantage that the process can be omitted and the process becomes simple.
松樹皮からの抽出は、上述の抽出法以外に、液体二酸化炭素回分法、液体二酸化炭素還流法、超臨界二酸化炭素還流法などにより行ってもよい。 Extraction from the pine bark may be performed by a liquid carbon dioxide batch method, a liquid carbon dioxide reflux method, a supercritical carbon dioxide reflux method, or the like, in addition to the above-mentioned extraction method.
松樹皮からの抽出は、複数の抽出方法を組み合わせてもよい。複数の抽出方法を組み合わせることにより、種々の組成の松樹皮抽出物を得ることが可能となる。 Extraction from pine bark may be a combination of a plurality of extraction methods. By combining a plurality of extraction methods, it becomes possible to obtain pine bark extracts having various compositions.
抽出により得られた松樹皮抽出物は、限外濾過、吸着性担体（ダイヤイオンＨＰ−２０、Ｓｅｐｈａｄｅｘ−ＬＨ２０、キチンなど）を用いたカラム法、バッチ法などにより精製を行うことが安全性の面から好ましい。 It is safe to purify the pine bark extract obtained by extraction by ultrafiltration, column method using adsorptive carrier (Diaion HP-20, Sephadex-LH20, chitin, etc.), batch method, etc. It is preferable from the aspect.
松樹皮抽出物は、保存性や加工性の観点から、粉末状のものであることが好ましく、乾燥粉末状のものであることがより好ましい。乾燥手段は特に限定されず、例えば、溶媒を含む松樹皮抽出物を、加温、日干し、熱風乾燥、凍結乾燥、減圧などによる乾燥手段を挙げることができる。乾燥の程度は、松樹皮抽出物の溶媒含有量が十分に低下したことが確認されるまでの程度であればよく、例えば、溶媒含有量が１０ｗｔ％以下、好ましくは５ｗｔ％以下となるまでの程度である。 The pine bark extract is preferably in the form of powder, more preferably in the form of dry powder, from the viewpoint of storage stability and processability. The drying means is not particularly limited, and examples thereof include drying means of the pine bark extract containing a solvent by heating, sun-drying, hot-air drying, freeze-drying, reduced pressure, and the like. The degree of drying may be such that the solvent content of the pine bark extract is sufficiently reduced, for example, until the solvent content is 10 wt% or less, preferably 5 wt% or less. Degree.
松樹皮抽出物乾燥粉末を得るための粉末化の方法としては、例えば、当業者が通常用いる方法であるボールミル、ハンマーミル、ローラーミルなどにより、松樹皮抽出物を粉砕及び粉末化する方法が挙げられるが、これらに限定されない。乾燥と粉末化の順序を入れ替えて、乾燥前の松樹皮抽出物を予め粉砕しておき、この粉砕物を乾燥して松樹皮抽出物乾燥粉末とすることもできる。 Examples of the powdering method for obtaining the pine bark extract dry powder include a method of pulverizing and pulverizing the pine bark extract by a ball mill, a hammer mill, a roller mill or the like which are usually used by those skilled in the art. However, it is not limited to these. It is also possible to change the order of drying and powdering to crush the pine bark extract before drying in advance and then dry the crushed product to obtain a pine bark extract dry powder.
松樹皮抽出物は市販されているものでもよく、市販の松樹皮抽出物としては、例えば、フラバンジェノール（登録商標；株式会社東洋新薬）などが挙げられる。 The pine bark extract may be commercially available, and examples of the commercially available pine bark extract include flavangenol (registered trademark; Toyo Shinyaku Co., Ltd.).
本発明の組成物は、プロシアニジンＢ３を含有する。プロアントシアニジンＢ３は、（＋）カテキンと（＋）カテキンとがＣ４−Ｃ８結合した二量体である。本発明のプロシアニジンＢ３（ＰＢ３）は、合成物や松、リンゴ、ココアやブドウなどの植物由来物を用いることができ、使用時の安全性の観点から植物由来物が好ましく、松がより好ましく、ＰＢ１を豊富に含むことから、松樹皮を用いることが特に好ましい。本発明の組成物に使用できる松の種類や植物由来物の加工方法は、ＰＢ１に記載した内容を参照できる。 The composition of the present invention contains procyanidin B3. Proanthocyanidin B3 is a dimer in which (+) catechin and (+) catechin are bound to C4-C8. As the procyanidin B3 (PB3) of the present invention, synthetic products and plant-derived products such as pine, apple, cocoa and grape can be used, and from the viewpoint of safety during use, plant-derived products are preferable, and pine is more preferable. It is particularly preferable to use pine bark because it contains abundant PB1. For the types of pine and the processing method of plant-derived products that can be used in the composition of the present invention, the contents described in PB1 can be referred to.
本発明の血管内皮機能改善用組成物はＰＢ１及びＰＢ３を含有することを特徴とする。本発明において、ＰＢ１とＰＢ３との含有比は特に限定されないが、血管内皮機能改善作用の効果を高める観点から、そのＰＢ１とＰＢ３の含有比がＰＢ１に対してＰＢ３が１．５以下であることが好ましく、ＰＢ３が０．７６以下であることがより好ましく、ＰＢ３が０．６８以下であることが特に好ましく、ＰＢ３が０．６０以下であることがより特に好ましい。また、ＰＢ３の含有比の下限は特に限定されないが、血管内皮機能改善作用の効果を高める観点から、ＰＢ１とＰＢ３の含有比がＰＢ１に対してＰＢ３が０．２０以上であることが好ましく、０．３６以上であることがより好ましく、０．４１以上であることが特に好ましい。 The composition for improving vascular endothelial function of the present invention is characterized by containing PB1 and PB3. In the present invention, the content ratio of PB1 and PB3 is not particularly limited, but the content ratio of PB1 and PB3 is 1.5 or less with respect to PB1 from the viewpoint of enhancing the effect of improving the vascular endothelial function. It is more preferable that PB3 is 0.76 or less, PB3 is particularly preferably 0.68 or less, and PB3 is more preferably 0.60 or less. Further, the lower limit of the content ratio of PB3 is not particularly limited, but from the viewpoint of enhancing the effect of improving the vascular endothelial function, the content ratio of PB1 and PB3 is preferably 0.20 or more with respect to PB1 and is 0. It is more preferably .36 or more, and particularly preferably 0.41 or more.
本発明の血管柔軟性向上用組成物はＰＢ１及びＰＢ３を含有することを特徴とする。本発明において、ＰＢ１とＰＢ３との含有比は特に限定されないが、血管柔軟性向上作用の効果を高める観点から、そのＰＢ１とＰＢ３の含有比がＰＢ１に対してＰＢ３が１．５０以下であることが好ましく、ＰＢ３が０．７６以下であることがより好ましく、ＰＢ３が０．６８以下であることが特に好ましく、ＰＢ３が０．６０以下であることがより特に好ましい。また、ＰＢ３の含有比の下限は特に限定されないが、血管柔軟性向上作用の効果を高める観点から、ＰＢ１とＰＢ３の含有比がＰＢ１に対してＰＢ３が０．２０以上であることが好ましく、ＰＢ３が０．３６以上であることがより好ましく、０．４１以上であることが特に好ましい。 The composition for improving vascular flexibility of the present invention is characterized by containing PB1 and PB3. In the present invention, the content ratio of PB1 and PB3 is not particularly limited, but the content ratio of PB1 and PB3 is 1.50 or less with respect to PB1 from the viewpoint of enhancing the effect of improving vascular flexibility. It is more preferable that PB3 is 0.76 or less, PB3 is particularly preferably 0.68 or less, and PB3 is more preferably 0.60 or less. Further, the lower limit of the content ratio of PB3 is not particularly limited, but from the viewpoint of enhancing the effect of improving the vascular flexibility, the content ratio of PB1 and PB3 is preferably 0.20 or more with respect to PB1 and PB3. Is more preferably 0.36 or more, and particularly preferably 0.41 or more.
本発明の血流依存性血管拡張機能改善用組成物はＰＢ１及びＰＢ３を含有することを特徴とする。本発明において、ＰＢ１とＰＢ３との含有比は特に限定されないが、血流依存性血管拡張機能改善作用の効果を高める観点から、ＰＢとＰＢ３の含有比の上限は、ＰＢ１に対してＰＢ３が１．３０以下であることが好ましく、ＰＢ３が０．７６以下であることがより好ましく、０．６８以下であることが特に好ましい。また、同様にＰＢ１とＰＢ３の含有比の下限は、血流依存性血管拡張機能改善作用の効果を高める観点から、ＰＢ１とＰＢ３の含有比がＰＢ１に対してＰＢ３が０．２０以上であることが好ましく、０．３６以上であることがより好ましく、０．４１以上であることが特に好ましい。 The composition for improving blood flow-dependent vasodilator function of the present invention is characterized by containing PB1 and PB3. In the present invention, the content ratio of PB1 and PB3 is not particularly limited, but from the viewpoint of enhancing the effect of improving the blood flow-dependent vasodilator function, the upper limit of the content ratio of PB and PB3 is 1 for PB1. It is preferably .30 or less, more preferably 0.76 or less, and particularly preferably 0.68 or less. Similarly, the lower limit of the content ratio of PB1 and PB3 is that the content ratio of PB1 and PB3 is 0.20 or more with respect to PB1 from the viewpoint of enhancing the effect of improving the blood flow-dependent vasodilator function. Is preferable, 0.36 or more is more preferable, and 0.41 or more is particularly preferable.
ＰＢ１及びＰＢ３を含有する組成物は、後述する実施例に示されているとおり、血流依存性血管拡張機能改善作用を示すことから、血管内皮機能改善作用、血管柔軟性向上作用などが期待できるものである、という態様を採り得る。 As shown in Examples described later, the composition containing PB1 and PB3 exhibits a blood flow-dependent vasodilatory function improving action, and thus can be expected to have a vascular endothelial function improving action, a vascular flexibility improving action, and the like. It is possible to take the aspect of being a thing.
本発明の組成物の１日の使用量は特に限定されず、使用態様や使用者の使用内容などに応じて適宜設定され得るが、例えば、１日摂取量当たりのＰＢ１とＰＢ３の合計量は、０．１〜１００ｍｇであり、好ましくは１ｍｇ〜５０ｍｇであり、血管内皮機能改善用組成物作用、血管柔軟性向上作用、及び血流依存性血管拡張機能改善作用の観点から、さらに好ましくは２〜１０ｍｇである。 The daily amount of the composition of the present invention is not particularly limited and may be appropriately set according to the usage mode and the content of use by the user. For example, the total amount of PB1 and PB3 per daily intake is , 0.1 to 100 mg, preferably 1 mg to 50 mg, and more preferably 2 from the viewpoint of the composition action for improving vascular endothelial function, the vascular flexibility improving action, and the blood flow-dependent vasodilator function improving action. It is 10 mg.
本発明の血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物は、ヒトに対して好適に適用されるものであるが、期待される作用効果が奏される限り特に限定はなく、ヒト以外の動物に対して適用することができる。本発明の松樹皮抽出物含有剤の使用者は特に限定されず、例えば、健常者であってもよいが、血管内皮のリスクファクターを有する者や４０歳以上の中高年者に用いることがより好ましい。本発明の血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物の使用頻度は特に限定されないが、好ましくは２日に１度以上、より好ましくは１日に１度以上、特に好ましくは１日に２度以上である。組成物中に含有する１回当たりのＰＢ１とＰＢ３の合計量は、０．１〜１００ｍｇであり、好ましくは１ｍｇ〜５０ｍｇであり、血管内皮機能改善用組成物作用、血管柔軟性向上作用、及び血流依存性血管拡張機能改善作用の観点から、さらに好ましくは２〜１０ｍｇである。 The composition for improving vascular endothelial function, the composition for improving vascular flexibility, and the composition for improving blood flow-dependent vasodilator function of the present invention are preferably applied to humans, but are expected. There is no particular limitation as long as the action and effect are exhibited, and it can be applied to animals other than humans. The user of the pine bark extract-containing agent of the present invention is not particularly limited, and may be, for example, a healthy person, but it is more preferable to use it for a person having a risk factor for vascular endothelium or a middle-aged person aged 40 years or older. .. The frequency of use of the composition for improving vascular endothelial function, the composition for improving vascular flexibility, and the composition for improving blood flow-dependent vasodilator function of the present invention is not particularly limited, but is preferably once or more every two days. It is preferably at least once a day, and particularly preferably at least twice a day. The total amount of PB1 and PB3 contained in the composition at one time is 0.1 to 100 mg, preferably 1 mg to 50 mg, and has an action of a composition for improving vascular endothelial function, an action of improving vascular flexibility, and an action of improving vascular flexibility. From the viewpoint of blood flow-dependent vasodilatory function improving action, it is more preferably 2 to 10 mg.
本発明の血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物におけるＰＢ１とＰＢ３の合計量の配合割合は、その投与形態や剤形などによって適宜設定することができ、特に限定されない。例えば、ＰＢ１とＰＢ３の合計量は、全体を１００質量部として、ＰＢ１とＰＢ３の合計量の下限値は乾燥質量で、０．００１質量部以上、好ましくは０．０１質量部以上、より好ましくは０．０５質量部以上と設定することができ、ＰＢ１とＰＢ３の合計量の上限値は乾燥質量で、例えば、１００質量部以下、好ましくは５０質量部以下、より好ましくは１０質量部以下と設定することができる。 The compounding ratio of the total amount of PB1 and PB3 in the composition for improving vascular endothelial function, the composition for improving vascular flexibility, and the composition for improving blood flow-dependent vasodilator function of the present invention is the administration form, dosage form, etc. It can be set as appropriate, and is not particularly limited. For example, the total amount of PB1 and PB3 is 100 parts by mass as a whole, and the lower limit of the total amount of PB1 and PB3 is the dry mass of 0.001 part by mass or more, preferably 0.01 part by mass or more, more preferably. It can be set to 0.05 parts by mass or more, and the upper limit of the total amount of PB1 and PB3 is set to dry mass, for example, 100 parts by mass or less, preferably 50 parts by mass or less, and more preferably 10 parts by mass or less. can do.
本発明の血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物における松樹皮抽出物の配合量は、その投与形態や剤形などによって適宜設定することができ、特に限定されない。例えば、松樹皮抽出物の配合量は、全体を１００質量部として、松樹皮抽出物の下限値は乾燥質量で、０．００１質量部以上、０．０１質量部以上、好ましくは０．１質量部以上と設定することができ、松樹皮抽出物の上限値は乾燥質量で、例えば、１００質量部以下、好ましくは５０質量部以下、より好ましくは３０質量部以下と設定することができる。 The blending amount of the pine bark extract in the composition for improving vascular endothelial function, the composition for improving vascular flexibility, and the composition for improving blood flow-dependent vasodilator function of the present invention is appropriately determined depending on the administration form and dosage form. It can be set and is not particularly limited. For example, the blending amount of the pine bark extract is 100 parts by mass as a whole, and the lower limit of the pine bark extract is the dry mass of 0.001 part by mass or more, 0.01 part by mass or more, preferably 0.1 mass by mass. The upper limit of the pine bark extract can be set to 100 parts by mass or less, preferably 50 parts by mass or less, and more preferably 30 parts by mass or less.
本発明の組成物は、ＰＢ１及びＰＢ３に加えて、適宜選択したその他の成分を含有してもよい。その他の成分としては、例えば、種々の賦形剤、結合剤、滑沢剤、安定剤、希釈剤、増量剤、増粘剤、乳化剤、着色料、香料、添加剤、化粧品原料、医薬品原料などを挙げることができる。その他の成分の含有量は、本発明の血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物の利用形態などに応じて適宜選択することができる。 In addition to PB1 and PB3, the composition of the present invention may contain other components appropriately selected. Other ingredients include, for example, various excipients, binders, lubricants, stabilizers, diluents, bulking agents, thickeners, emulsifiers, colorants, fragrances, additives, cosmetic raw materials, pharmaceutical raw materials, etc. Can be mentioned. The content of other components should be appropriately selected according to the usage form of the composition for improving vascular endothelial function, the composition for improving vascular flexibility, and the composition for improving blood flow-dependent vasodilator function of the present invention. Can be done.
本発明の組成物は、血管内皮機能改善作用、血管柔軟性向上作用、若しくは血流依存性血管拡張機能改善作用を得ることを目的とした種々の形態で利用することができ、例えば、経口用又は非経口用の形態とすることができる。本発明の組成物は、その形態に応じて、そのまま経口的又は非経口的に使用してもよいし、松樹皮抽出物を溶解するための溶媒などに溶解して経口的又は非経口的に使用してもよいが、本発明の効果である血管内皮機能改善作用、血管柔軟性向上作用、及び血流依存性血管拡張機能改善作用をより効果的に発揮できる観点から、経口が好ましい。 The composition of the present invention can be used in various forms for the purpose of obtaining a vascular endothelial function improving action, a vascular flexibility improving action, or a blood flow-dependent vasodilator function improving action, for example, for oral use. Alternatively, it can be in the form for parenteral use. The composition of the present invention may be used orally or parenterally as it is, depending on its form, or it may be dissolved orally or parenterally in a solvent for dissolving the pine bark extract. Although it may be used, oral administration is preferable from the viewpoint of more effectively exerting the effects of the present invention, such as the effect of improving vascular endothelial function, the effect of improving vascular flexibility, and the effect of improving blood flow-dependent vasodilator function.
本発明の組成物の形態が経口用である場合、その形態としては、経口的な使用に適した形態であれば良く、例えば、顆粒状、粉末状、錠状、チュアブル状、カプセル状、液状、シロップ状などが挙げられる。これらの中でも、ＰＢ１及びＰＢ３の効率的な摂取の観点から、錠状が好ましい。 When the form of the composition of the present invention is for oral use, the form may be any form suitable for oral use, for example, granules, powders, tablets, chewables, capsules, and liquids. , Syrup-like, etc. Among these, a tablet shape is preferable from the viewpoint of efficient ingestion of PB1 and PB3.
また、本発明の組成物としては、有効成分を含有する血管内皮機能改善用食品の他、食品に対して有効成分を添加して得た血管内皮機能改善用食品を挙げることができ、例えば、通常の食品に比して本発明の有効成分含有量を増加させた食品や、本発明の有効成分を通常含まない食品に対して有効成分を添加した食品を挙げることができる。有効成分の添加は、それぞれの成分を別々に添加してもよいし、同時に添加してもよく、また、有効成分以外の他の成分と共に添加してもよい。 Further, examples of the composition of the present invention include foods for improving vascular endothelial function containing an active ingredient, and foods for improving vascular endothelial function obtained by adding an active ingredient to a food, for example. Examples thereof include foods in which the content of the active ingredient of the present invention is increased as compared with ordinary foods, and foods in which the active ingredient is added to foods that do not normally contain the active ingredient of the present invention. The active ingredient may be added separately, simultaneously, or together with other ingredients other than the active ingredient.
また、本発明の組成物の形態が非経口用である場合、その形態としては、例えば、非経口的な使用に適した形態、具体的には、ローション状、クリーム状、リキッド状、ミスト状、エマルション状、スプレー状、ムース状、ジェル状などが挙げられる。 When the form of the composition of the present invention is for parenteral use, the form thereof is, for example, a form suitable for parenteral use, specifically, a lotion-like, cream-like, liquid-like, or mist-like form. , Emulsion, spray, mousse, gel and the like.
本発明の組成物の製造方法は特に限定されず、使用形態に応じて当業者に知られる一般的な製造方法に準じて製造される。例えば、顆粒状や固形状である場合、ＰＢ１及びＰＢ３をそのまま又は上記のその他の成分と同時又は数段階に分けて混和したものを、流動層造粒法、攪拌造粒法、押出造粒法などの造粒方法に従って造粒して顆粒状とし、さらに打錠機などを用いる常法に従って圧縮成形することによって錠状に成形できる。また、液状やクリーム状の場合は、ＰＢ１及びＰＢ３を上記のその他の成分と同時又は数段階に分けて混合し、調整することができる。 The method for producing the composition of the present invention is not particularly limited, and the composition is produced according to a general production method known to those skilled in the art depending on the mode of use. For example, in the case of granules or solids, PB1 and PB3 are mixed as they are or with the above other components at the same time or in several steps, and the fluidized bed granulation method, stirring granulation method, and extrusion granulation method are used. It can be formed into a tablet shape by granulating according to a granulation method such as, and then compression molding according to a conventional method using a tableting machine or the like. In the case of liquid or cream, PB1 and PB3 can be mixed with the above other components at the same time or in several steps to adjust.
本発明の組成物は、血管内皮機能改善、血管柔軟性向上、若しくは血流依存性血管拡張機能改善に用いられる点において、製品として他の製品と区別することができるものであれば特に制限されるものではなく、例えば、本発明に係る製品の本体、包装、説明書、宣伝物のいずれかに血管内皮機能改善作用の機能がある旨を表示したものが本発明の範囲に含まれる。例えば、医薬品（医薬部外品を含む）や、特定保健用食品、栄養機能食品、機能性表示食品等の所定機関より効能の表示が認められた機能性食品などのいわゆる健康食品や、飼料等を挙げることができる。いわゆる健康食品においては、「血管機能を高める」、「血管機能を維持する」、「血管内皮機能を高める」、「血管内皮機能を維持する」、「加齢によって衰える動脈の血管内皮機能を高める」、「加齢によって衰える動脈の血管内皮機能を維持する」、「血管を締め付けた後の血管の拡張度を高める」、「血管を締め付けた後の血管の拡張度を維持する」、「血管柔軟性を高める」、「血管柔軟性を維持する」、「血管のしなやかさを高める」、「血管のしなやかさを維持する」等を表示したものを例示することができる。 The composition of the present invention is particularly limited as long as it can be distinguished from other products as a product in that it is used for improving vascular endothelial function, improving vascular flexibility, or improving blood flow-dependent vasodilatory function. However, the scope of the present invention includes, for example, a product indicating that any of the main body, packaging, instruction manual, and advertising material of the product according to the present invention has a function of improving vascular endothelial function. For example, so-called health foods such as pharmaceuticals (including quasi-drugs), foods for specified health use, foods with nutritional function, foods with functional claims, etc. Can be mentioned. In so-called health foods, "enhance vascular function", "maintain vascular function", "enhance vascular endothelial function", "maintain vascular endothelial function", "enhance vascular endothelial function of arteries that decline with age" "Maintaining the vascular endothelial function of arteries that decline with age", "Increasing the degree of dilation of blood vessels after tightening blood vessels", "Maintaining the degree of dilation of blood vessels after tightening blood vessels", "Vascular vessels" Examples of those displaying "increasing flexibility", "maintaining vascular flexibility", "increasing vascular suppleness", "maintaining vascular suppleness", etc. can be exemplified.
本発明の別の態様は、ＰＢ１及びＰＢ３を含有する組成物、又は、ＰＢ１及びＰＢ３を含有する松樹皮抽出物を含有する組成物を使用させることを含む、血管内皮機能、血管柔軟性、若しくは血流依存性血管拡張機能を改善、又は維持する方法である。ただし、本発明の方法は、ヒトに対する医療行為を除外するものである。 Another aspect of the invention comprises the use of a composition containing PB1 and PB3, or a composition containing a pine bark extract containing PB1 and PB3, including vascular endothelial function, vascular flexibility, or. A method of improving or maintaining blood flow-dependent vasodilatory function. However, the method of the present invention excludes medical practice for humans.
本発明の血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物の包装形態は特に限定されず、剤形などに応じて適宜選択できるが、例えば、ＰＴＰなどのブリスターパック；ストリップ包装；ヒートシール；アルミパウチ；プラスチックや合成樹脂などを用いるフィルム包装；バイアルなどのガラス容器；アンプルなどのプラスチック容器などが挙げられる。 The packaging form of the composition for improving vascular endothelial function, the composition for improving vascular flexibility, and the composition for improving blood flow-dependent vasodilatory function of the present invention is not particularly limited and may be appropriately selected depending on the dosage form and the like. For example, blister packs such as PTP; strip packaging; heat seals; aluminum pouches; film packaging using plastic or synthetic resin; glass containers such as vials; plastic containers such as ampoules.
以下、本発明を実施例によりさらに詳細に説明するが、本発明はこれら実施例に限定されるものではなく、本発明の課題を解決し得る限り、本発明は種々の態様をとることができる。 Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples, and the present invention can take various aspects as long as the problems of the present invention can be solved. ..
＜試験１ 血流依存性血管拡張機能の評価＞
（１）被験者と試験デザイン
倫理的配慮により試験開始前に同意が得られた４０歳以上、６５歳未満の健常な成人男女３５名で、ＢＭＩが３０未満の人を被験者として選定した。被験者を無作為の２群（実施例群：１８名、比較例群：１７名）に分け、二重盲検法による並行群間比較試験をした。１日摂取目安量である１袋（錠剤０．２５ｇ×２粒）の試験食品を１日１回、水またはぬるま湯とともに摂取させ、８週間摂取後に検査を行った。
<Test 1 Evaluation of blood flow-dependent vasodilatory function>
(1) Subjects and study design Thirty-five healthy adult men and women aged 40 years or older and under 65 years of age who agreed before the start of the study due to ethical considerations were selected as subjects with a BMI of less than 30. Subjects were divided into two randomized groups (Example group: 18 subjects, Comparative example group: 17 subjects), and a double-blind, parallel-group comparative study was conducted. One bag (0.25 g of tablets x 2 tablets) of the test food, which is the recommended daily intake, was ingested once a day with water or lukewarm water, and the test was performed after 8 weeks of ingestion.
（２）試験食品
実施例１には、熱水で抽出されたフランス海岸松の松樹皮抽出物に、粉末還元麦芽糖水飴、微結晶セルロース、ショ糖脂肪酸エステル、微粒二酸化ケイ素を混合し、打錠した錠剤を使用した。比較例１には、外観で実施例との区別がつかないようにするため、実施例中の松樹皮抽出物をカラメル色素に置き換えたものを使用した。実施例１及び比較例１ともに１日摂取目安量（０．２５ｇ×２粒）を無地アルミ個包装し、被験者及び介入実施者への盲検性を確保した。試験食品の１日摂取目安量あたりの熱量及び栄養成分の分析結果を表１に示す。
(2) In Example 1 of the test food, powdered reduced maltose starch syrup, microcrystalline cellulose, sucrose fatty acid ester, and fine-grained silicon dioxide were mixed with the pine bark extract of French coastal pine extracted with hot water and tableted. The tablets used were used. In Comparative Example 1, in order to make it indistinguishable from the examples in appearance, the pine bark extract in the examples was replaced with a caramel color. In both Example 1 and Comparative Example 1, the recommended daily intake (0.25 g x 2 tablets) was individually wrapped in plain aluminum to ensure blindness to the subjects and the intervention performers. Table 1 shows the analysis results of the calorific value and nutritional components per daily intake standard amount of the test food.
（３）ＰＢ１及びＰＢ３含有量の分析
ＰＢ１及びＰＢ３量を以下の方法により測定した。松樹皮抽出物の重量を精密に測り、１０％エタノールを加えてメスアップし、口径０．４５μｍのメンブランフィルターで濾過し、試験溶液を得た。下記の条件でＨＰＬＣに供し、ＰＢ１、ＰＢ３の定量を行った。松樹皮抽出物に含まれるＰＢ１及びＰＢ３を測定し、実施例１（０．２５ｇ×２粒中；１日摂取目安量）に含まれるＰＢ１及びＰＢ３を算定した。実施例１に含まれるＰＢ１は１．７ｍｇであり、ＰＢ３は０．７ｍｇであり、ＰＢ１とＰＢ３の合計は２．４ｍｇであった。ＰＢ１に対するＰＢ３の比率は、０．４１であった。
（ＨＰＬＣ条件）
・分析カラム：Ｌ−Ｃｏｌｕｍｎ ＯＤＳ ３μｍ、４．６×２５０ｍｍ（一般財団法人化学物質評価研究機構製）
・カラム温度：４０℃
・注入量 ：１０μＬ
・流速 ：１．０ｍｌ／ｍｉｎ
・測定波長 ：２８０ｎｍ
・移動相 ：（Ａ液）０．１Ｍ酢酸水溶液
（Ｂ液）０．１Ｍ酢酸アセトニトリル溶液
・グラジエントプログラム：表２参照
(3) Analysis of PB1 and PB3 contents The amounts of PB1 and PB3 were measured by the following methods. The pine bark extract was weighed precisely, 10% ethanol was added, the volumetric flask was increased, and the mixture was filtered through a membrane filter having a diameter of 0.45 μm to obtain a test solution. It was subjected to HPLC under the following conditions, and PB1 and PB3 were quantified. PB1 and PB3 contained in the pine bark extract were measured, and PB1 and PB3 contained in Example 1 (0.25 g × 2 grains; daily intake guideline amount) were calculated. The amount of PB1 contained in Example 1 was 1.7 mg, the amount of PB3 was 0.7 mg, and the total of PB1 and PB3 was 2.4 mg. The ratio of PB3 to PB1 was 0.41.
(HPLC conditions)
-Analytical column: L-Column ODS 3 μm, 4.6 x 250 mm (manufactured by Chemicals Evaluation and Research Institute)
-Column temperature: 40 ° C
・ Injection amount: 10 μL
・ Flow velocity: 1.0 ml / min
-Measurement wavelength: 280 nm
-Mobile phase: (Liquid A) 0.1 M aqueous acetic acid solution
(Liquid B) 0.1 M Acetonitrile acetate solution-Gradient program: See Table 2.
（４）ＦＭＤ値の測定
血流依存性血管拡張機能は、超音波検査機器を用いたＦＭＤ（ｆｌｏｗ−ｍｅｄｉａｔｅｄ ｄｉｌａｔｉｏｎ）により有効性評価をした。具体的には上腕部の安静時動脈血管径を計測した後、前腕部をカフで５分間圧迫し、駆血解放後の最大動脈血管径を計測した。ＦＭＤ値は最大動脈血管径と安静時動脈血管径の差を、安静時動脈血管径で除し、百分率で算出した。結果を表３に示す。
(4) Measurement of FMD value The effectiveness of the blood flow-dependent vasodilator function was evaluated by FMD (flow-mediated dilation) using an ultrasonic inspection device. Specifically, after measuring the resting arterial blood vessel diameter of the upper arm, the forearm was pressed with a cuff for 5 minutes, and the maximum arterial blood vessel diameter after the release of avascularization was measured. The FMD value was calculated as a percentage by dividing the difference between the maximum arterial blood vessel diameter and the resting arterial blood vessel diameter by the resting arterial blood vessel diameter. The results are shown in Table 3.
二元配置分散分析において交互作用が認められ（ｐ＜０．０５）、摂取８週間後の実測値及び摂取前から摂取８週間後の変化量において、実施例１の群は比較例１の群と比較して有意な増加が認められた。 An interaction was observed in the two-way ANOVA (p <0.05), and the group of Example 1 was the group of Comparative Example 1 in terms of the measured values 8 weeks after ingestion and the amount of change from before ingestion to 8 weeks after ingestion. A significant increase was observed in comparison with.
以上より、本試験の結果、ＰＢ１及びＰＢ３を含有する錠剤を摂取することによって、ＦＭＤ値は増加し、血流依存性血管拡張機能の改善が確認された。したがって、本発明の組成物は血流依存性血管拡張機能改善による優れた血管柔軟性向上効果、血管内皮機能改善効果を有することがわかる。特に、ＰＢ１とＰＢ３を合計で１日当たり２．４ｍｇ配合した錠剤を摂取することで、優れた血流依存性血管拡張機能の改善効果が見られることから、本発明の組成物は、ＰＢ１とＰＢ３を合計で１日当たり２〜１０ｍｇ摂取することで、血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果が顕著に発揮されることが示唆された。 From the above, as a result of this test, it was confirmed that the FMD value was increased and the blood flow-dependent vasodilator function was improved by ingesting the tablets containing PB1 and PB3. Therefore, it can be seen that the composition of the present invention has an excellent effect of improving vascular flexibility and an effect of improving vascular endothelial function by improving blood flow-dependent vasodilatory function. In particular, the composition of the present invention has PB1 and PB3 because an excellent effect of improving blood flow-dependent vasodilatory function can be seen by ingesting a tablet containing a total of 2.4 mg of PB1 and PB3 per day. It was suggested that the effect of improving vascular endothelial function, the effect of improving vascular flexibility, and the effect of improving blood flow-dependent vasodilatory function were remarkably exhibited by ingesting 2 to 10 mg of vasodilator per day in total.
＜試験２ 酸化ストレス下での活性酸素種（ＲＯＳ）産生量の評価＞
本試験では、酸化ストレス下での活性酸素種（ＲＯＳ）産生量について評価した。酸化ストレスによって増加したＲＯＳはＮＯを酸化し、酸化されたＮＯは生理活性を低下させる。酸化ストレス下でのＲＯＳ産生量の抑制は、ＮＯの酸化による生理活性の低下を防ぎ、ＮＯ利用能の改善につながるため、血管内皮機能改善、血管柔軟性向上、血流依存性血管拡張機能改善が期待できる。以上より、酸化ストレス下でのＲＯＳ産生量を評価することで、血管内皮機能改善作用、血管柔軟性向上作用及び、血流依存性血管拡張改善作用の評価ができる。
<Test 2 Evaluation of reactive oxygen species (ROS) production under oxidative stress>
In this test, the amount of reactive oxygen species (ROS) produced under oxidative stress was evaluated. ROS increased by oxidative stress oxidizes NO, and oxidized NO reduces bioactivity. Suppression of ROS production under oxidative stress prevents a decrease in physiological activity due to NO oxidation and leads to improvement of NO utilization, so improvement of vascular endothelial function, improvement of vascular flexibility, and improvement of blood flow-dependent vasodilator function. Can be expected. From the above, by evaluating the amount of ROS produced under oxidative stress, it is possible to evaluate the vascular endothelial function improving action, the vascular flexibility improving action, and the blood flow-dependent vasodilation improving action.
被験物質
ＰＢ１及びＰＢ３の量が表４の割合となるように、熱水で抽出されたフランス海岸松の松樹皮抽出物をＤＭＳＯで溶解し、添加濃度でＤＭＳＯが０．５％になるようにＥｎｄｏｔｈｅｌｉａｌ Ｃｅｌｌ Ｇｒｏｗｔｈ Ｍｅｄｉｕｍ ２（ＥＧＭ−２）培地（ＬＯＮＺＡ製）で希釈することで、実施例及び比較例の被験物質を調製した。なお、ＰＢ１及びＰＢ３の量は、「試験１（３）ＰＢ１及びＰＢ３含有量の分析」に記載される方法と同様の方法で測定した。
The pine bark extract of French coastal pine extracted with hot water was dissolved in DMSO so that the amounts of the test substances PB1 and PB3 were in the ratio shown in Table 4, and the DMSO was 0.5% at the added concentration. Test substances of Examples and Comparative Examples were prepared by diluting with Endothelial Cell Growth Medium 2 (EGM-2) medium (manufactured by LONZA). The amounts of PB1 and PB3 were measured by the same method as described in "Test 1 (3) Analysis of PB1 and PB3 contents".
（２）細胞培養と酸化ストレス
ヒト臍帯静脈内皮細胞（ＨＵＶＥＣ）をコラーゲンコートされた９６ウェルのブラックプレート（ＲＯＳ産生量測定用）とクリアプレート（細胞増殖試験用）に８．０×１０３ｃｅｌｌｓ／ｗｅｌｌとなるように播種し、３７℃、５％ＣＯ２インキュベーター内で、２４時間培養した。培地を除去後、被験物質含有培地を１００μＬ／ｗｅｌｌ添加し、３７℃、５％ＣＯ２インキュベーター内で２２時間培養した。その後、過酸化水素（ナカライテスク製）を終濃度が１００μＭとなるように添加することで、酸化ストレスを与えた。その後、３７℃、５％ＣＯ２インキュベーター内で、２時間培養した後に、ＲＯＳ産生量の測定と細胞増殖試験を行った。
(2) Cell culture and oxidative stress 8.0 x 10 3 cells on a 96-well black plate (for measuring ROS production) and a clear plate (for cell proliferation test) coated with collagen of human umbilical vein endothelial cells (HUVEC). The cells were sown at / well and cultured at 37 ° C. in a 5% CO 2 incubator for 24 hours. After removing the medium, 100 μL / well of the test substance-containing medium was added, and the cells were cultured at 37 ° C. in a 5% CO 2 incubator for 22 hours. Then, hydrogen peroxide (manufactured by Nacalai Tesque) was added so as to have a final concentration of 100 μM to give oxidative stress. Then, after culturing in a 5% CO 2 incubator at 37 ° C. for 2 hours, the ROS production amount was measured and a cell proliferation test was performed.
（３）ＲＯＳ産生量の測定
ブラックプレート（ＲＯＳ産生量測定用）に関して、培地を除去後、無血清ＤＭＥＭ培地で２回洗浄し、１０μＭのＤＣＦＨ−ＤＡ試薬を５０μＬ／ｗｅｌｌ 添加し、３７℃、５％ＣＯ２インキュベーター内で３０分間培養した。培地を除去後、無血清ＤＭＥＭ培地で２回洗浄し、ＰＢＳを２００μｌ／ｗｅｌｌ添加した。４８５ｎｍで励起したときの５３０ｎｍにおける蛍光強度をプレートリーダー（Ｍｏｌｅｃｕｌａｒ Ｄｅｖｉｃｅｓ製）測定し、ＲＯＳ産生量を算出した。
(3) Measurement of ROS production amount With respect to the black plate (for measuring ROS production amount), after removing the medium, the medium was washed twice with serum-free DMEM medium, 50 μL / well of 10 μM DCFH-DA reagent was added, and 37 ° C. Incubated in 5% CO 2 incubator for 30 minutes. After removing the medium, the cells were washed twice with serum-free DMEM medium, and PBS was added at 200 μl / well. The fluorescence intensity at 530 nm when excited at 485 nm was measured by a plate reader (manufactured by Molecular Devices), and the amount of ROS produced was calculated.
（４）細胞増殖率の測定
クリアプレート（細胞増殖試験用）に関して、培地を除去後、無血清ＤＭＥＭ培地で２回洗浄し、Ｃｅｌｌ ｃｏｕｎｔｉｎｇ ｋｉｔ−８溶液を１５０μＬ／ｗｅｌｌ添加した。インキュベーター内で適度に発色させ、４５０ｎｍにおける吸光度をプレートリーダー（Ｍｏｌｅｃｕｌａｒ Ｄｅｖｉｃｅｓ製）で測定した。
(4) Measurement of cell proliferation rate With respect to the clear plate (for cell proliferation test), the medium was removed, washed twice with serum-free DMEM medium, and 150 μL / well of Cell counting kit-8 solution was added. The color was appropriately developed in the incubator, and the absorbance at 450 nm was measured with a plate reader (manufactured by Molecular Devices).
（５）ＲＯＳ産生量の評価
ＲＯＳ産生量を細胞増殖率で割り戻すことで、細胞数当たりのＲＯＳ産生量を算出し、酸化ストレスを与え、被験物質を投与しなかった群（比較例２）の細胞数当たりのＲＯＳ産生量を１００とした場合の、相対的なＲＯＳ産生量を算出した。結果を図１に示す。
(5) Evaluation of ROS production amount By dividing the ROS production amount by the cell proliferation rate, the ROS production amount per cell number was calculated, oxidative stress was applied, and the test substance was not administered (Comparative Example 2). The relative ROS production amount was calculated when the ROS production amount per cell number was 100. The results are shown in FIG.
被験物質を投与せず、酸化ストレスを与えなかった群（比較例３）に比べて、被験物質を投与せず、酸化ストレスを与えた群（比較例２）はＲＯＳ産生量が極めて高いことから、酸化ストレスによって、ＲＯＳが産生されることが確認された。比較例２及び実施例２〜７との比較から、ＰＢ１及びＰＢ３を含有する被験物質を投与した群（実施例２〜７）において、酸化ストレス下でのＲＯＳ産生量の抑制が認められた。したがって、ＰＢ１とＰＢ３を含有する組成物を摂取することにより、酸化ストレス下でのＲＯＳ産生量の抑制効果が発揮されることで、血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果が発揮されることが分かった。特に、ＰＢ１に対するＰＢ３の比が０．３６〜１．３０である被験物質を投与した群（実施例２〜６）は、酸化ストレス下でのＲＯＳ産生量の抑制に優れており、ＰＢ１に対するＰＢ３の比が０．４１〜０．６８である被験物質を投与した群（実施例３〜５）は、酸化ストレス下でのＲＯＳ産生量の抑制により優れており、ＰＢ１に対するＰＢ３の比が０．４１〜０．６０である被験物質を投与した群（実施例３、４）は、酸化ストレス下でのＲＯＳ産生量の抑制に極めて優れていることが認められた。このことから、本発明の組成物はＰＢ１及びＰＢ３を含有することにより、優れた血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果が発揮され、ＰＢ１に対するＰＢ３の比が０．３６〜１．３０である場合には、血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果が顕著に発揮され、ＰＢ１に対するＰＢ３の比が０．４１〜０．６８である場合には、血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果がより顕著に発揮され、ＰＢ１に対するＰＢ３の比が０．４１〜０．６０である場合には、血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果が極めて顕著に発揮されることが分かった。 Compared to the group to which the test substance was not administered and oxidative stress was not given (Comparative Example 3), the group to which the test substance was not administered and was given oxidative stress (Comparative Example 2) had an extremely high ROS production amount. , It was confirmed that ROS is produced by oxidative stress. From the comparison with Comparative Example 2 and Examples 2 to 7, suppression of ROS production under oxidative stress was observed in the group to which the test substance containing PB1 and PB3 was administered (Examples 2 to 7). Therefore, by ingesting the composition containing PB1 and PB3, the effect of suppressing the amount of ROS produced under oxidative stress is exhibited, so that the effect of improving vascular endothelial function, the effect of improving vascular flexibility, and the dependence on blood flow are exhibited. It was found that the effect of improving vasodilatory function was exhibited. In particular, the group (Examples 2 to 6) to which the test substance having a ratio of PB3 to PB1 of 0.36 to 1.30 was administered was excellent in suppressing the amount of ROS produced under oxidative stress, and PB3 to PB1. The group to which the test substance having a ratio of 0.41 to 0.68 was administered (Examples 3 to 5) was superior in suppressing the amount of ROS produced under oxidative stress, and the ratio of PB3 to PB1 was 0. It was found that the group to which the test substance of 41 to 0.60 was administered (Examples 3 and 4) was extremely excellent in suppressing the amount of ROS produced under oxidative stress. From this, by containing PB1 and PB3, the composition of the present invention exhibits an excellent effect of improving vascular endothelial function, an effect of improving vascular flexibility, and an effect of improving blood vessel-dependent vasodilatory function, and PB3 with respect to PB1. When the ratio is 0.36 to 1.30, the vascular endothelial function improving effect, the vascular flexibility improving effect, and the blood vessel-dependent vasodilator function improving effect are remarkably exhibited, and the ratio of PB3 to PB1 is 0. When it is 41 to 0.68, the effect of improving vascular endothelial function, the effect of improving vascular flexibility, and the effect of improving blood vessel-dependent vasodilatory function are more prominently exhibited, and the ratio of PB3 to PB1 is 0.41 to 0. When it was .60, it was found that the effect of improving the vascular endothelial function, the effect of improving the vascular flexibility, and the effect of improving the blood vessel-dependent vasodilatory function were remarkably exhibited.
＜試験３ ＮＯ合成酵素遺伝子発現量の評価＞
本試験では、ＮＯ合成酵素遺伝子発現量について評価した。血管内皮でＮＯ合成酵素（ｅＮＯＳ）によりＮＯが産生し、拡散されて血管平滑筋に届くと、筋肉が弛緩されて血管が拡張する。ＮＯ合成酵素遺伝子発現量が増加すると、ＮＯ合成酵素量が増加し、ＮＯ産生量が増加するため、血管内皮機能改善、血管柔軟性向上、血流依存性血管拡張機能改善が期待できる。以上より、ＮＯ合成酵素遺伝子発現量を評価することで、血管内皮機能改善作用、血管柔軟性向上作用及び、血流依存性血管拡張改善作用の評価ができる。
<Test 3 Evaluation of NO synthase gene expression level>
In this test, the expression level of NO synthase gene was evaluated. When NO is produced in the vascular endothelium by NO synthase (eNOS) and diffused to reach vascular smooth muscle, the muscle is relaxed and the blood vessel dilates. When the expression level of the NO synthase gene increases, the amount of NO synthase increases and the amount of NO production increases. Therefore, improvement of vascular endothelial function, improvement of vascular flexibility, and improvement of blood flow-dependent vasodilatory function can be expected. From the above, by evaluating the expression level of the NO synthase gene, it is possible to evaluate the vascular endothelial function improving action, the vascular flexibility improving action, and the blood flow-dependent vasodilation improving action.
（１）被験物質
ＰＢ１及びＰＢ３の量が表５の割合となるように、熱水で抽出されたフランス海岸松の松樹皮抽出物をＤＭＳＯで溶解し、添加濃度でＤＭＳＯが０．５％になるようにＥｎｄｏｔｈｅｌｉａｌ Ｃｅｌｌ Ｇｒｏｗｔｈ Ｍｅｄｉｕｍ ２（ＥＧＭ−２）培地（ＬＯＮＺＡ製）で希釈することで、実施例及び比較例の被験物質を調製した。なお、ＰＢ１及びＰＢ３の量は、「試験１（３）ＰＢ１及びＰＢ３含有量の分析」に記載される方法と同様の方法で測定した。
(1) Dissolve the pine bark extract of French coastal pine extracted with hot water in DMSO so that the amounts of the test substances PB1 and PB3 are in the ratio shown in Table 5, and the DMSO is 0.5% at the addition concentration. The test substances of Examples and Comparative Examples were prepared by diluting with Endothelial Cell Growth Medium 2 (EGM-2) medium (manufactured by LONZA) so as to be. The amounts of PB1 and PB3 were measured by the same method as described in "Test 1 (3) Analysis of PB1 and PB3 contents".
（２）細胞培養
ヒト臍帯静脈内皮細胞（ＨＵＶＥＣ）をコラーゲンコートされた２４ウェルのプレートに５．０×１０４ｃｅｌｌｓ／ｗｅｌｌとなるように播種し、３７℃、５％ＣＯ２インキュベーター内で２４時間培養した。培地を除去後、被験物質含有培地を５００μＬ／ｗｅｌｌ添加し、３７℃、５％ＣＯ２インキュベーター内で２４時間培養した。
(2) Cell culture Human umbilical vein endothelial cells (HUVEC) were seeded on a collagen-coated 24-well plate at 5.0 × 10 4 cells / well, and 24 in a 5% CO 2 incubator at 37 ° C. Incubated for hours. After removing the medium, 500 μL / well of the test substance-containing medium was added, and the cells were cultured at 37 ° C. in a 5% CO 2 incubator for 24 hours.
（３）ＮＯ合成酵素遺伝子発現量の測定
培地を除去後、ＰＢＳで２回洗浄した。ＲＮｅａｓｙ Ｍｉｎｉ Ｋｉｔ（ＱＩＡＧＥＮ製）を用いてＲＮＡを回収し、ＲｅｖｅｒＴｒａ Ａｃｅ（登録商標） ｑＰＣＲ ＲＴ Ｍａｓｔｅｒ Ｍｉｘ ｗｉｔｈ ｇＤＮＡ Ｒｅｍｏｖｅｒ（ＴＯＹＯＢＯ製）を用いてｃＤＮＡを合成した。得られたｃＤＮＡを鋳型として、ｅＮＯＳ遺伝子のプライマー（ＴＡＫＡＲＡ製）を用いて、Ｒｏｔｏｒ−Ｇｅｎｅ ＳＹＢＲ Ｇｒｅｅｎ ＰＣＲ Ｋｉｔ（ＱＩＡＧＥＮ製）により定量リアルタイムＰＣＲを行い、ｅＮＯＳ遺伝子のｍＲＮＡ発現量を測定した。内在性コントロールとして、βＡｃｔｉｎのプライマー（ＱＩＡＧＥＮ製）を用いて、βＡｃｔｉｎのｍＲＮＡ発現量を測定した。
(3) Measurement of NO synthase gene expression level After removing the medium, the cells were washed twice with PBS. RNA was recovered using RNeasy Mini Kit (manufactured by QIAGEN), and cDNA was synthesized using RiverTra Ace® qPCR RT Master Mix with gDNA Remover (manufactured by TOYOBO). Using the obtained cDNA as a template, quantitative real-time PCR was performed by Rotor-Gene SYBR Green PCR Kit (manufactured by QIAGEN) using an eNOS gene primer (manufactured by TAKARA), and the mRNA expression level of the eNOS gene was measured. As an endogenous control, βActin mRNA expression level was measured using a βActin primer (manufactured by QIAGEN).
（５）ＮＯ合成酵素遺伝子発現量の評価
被験物質を投与しなかった群（比較例４）の発現量を１とした場合の、ＮＯ合成酵素遺伝子発現量の相対値を算出した。結果を図２に示す。
(5) Evaluation of NO synthase gene expression level The relative value of the NO synthase gene expression level was calculated when the expression level of the group to which the test substance was not administered (Comparative Example 4) was set to 1. The results are shown in FIG.
ＰＢ１及びＰＢ３を含有する被験物質を投与した群（実施例８〜１２）において、被験物質を投与しなかった群（比較例４）と比べて、ＮＯ合成酵素遺伝子発現量の増加が認められた。したがって、ＰＢ１及びＰＢ３を含有する組成物を摂取することにより、ＮＯ合成酵素遺伝子発現量の増加効果が発揮されることで、血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果が発揮されることが分かった。特に、ＰＢ１に対するＰＢ３の比が０．４１〜０．６８である被験物質を投与した群（実施例９〜１１）は、ＮＯ合成酵素遺伝子発現量の増加により優れており、ＰＢ１に対するＰＢ３の比が０．４１〜０．６０である被験物質を投与した群（実施例９、１０）は、ＮＯ合成酵素遺伝子発現量の増加効果に極めて優れていることが認められた。このことから、本発明の組成物はＰＢ１及びＰＢ３を含有することにより、優れた血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果が発揮され、ＰＢ１に対するＰＢ３の比が０．４１〜０．６８である場合には、血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果がより顕著に発揮され、ＰＢ１に対するＰＢ３の比が０．４１〜０．６０である場合には、血管内皮機能改善効果、血管柔軟性向上効果、血流依存性血管拡張機能改善効果が極めて顕著に発揮されることが分かった。 In the group to which the test substance containing PB1 and PB3 was administered (Examples 8 to 12), an increase in the expression level of the NO synthase gene was observed as compared with the group to which the test substance was not administered (Comparative Example 4). .. Therefore, by ingesting the composition containing PB1 and PB3, the effect of increasing the expression level of the NO synthase gene is exhibited, so that the effect of improving the vascular endothelial function, the effect of improving the vascular flexibility, and the blood flow-dependent vasodilation are exhibited. It was found that the function improvement effect was exhibited. In particular, the group (Examples 9 to 11) to which the test substance having a ratio of PB3 to PB1 of 0.41 to 0.68 was administered was superior in the increase in the expression level of the NO synthase gene, and the ratio of PB3 to PB1. It was found that the group to which the test substance having a value of 0.41 to 0.60 was administered (Examples 9 and 10) was extremely excellent in the effect of increasing the expression level of the NO synthase gene. From this, by containing PB1 and PB3, the composition of the present invention exhibits an excellent effect of improving vascular endothelial function, an effect of improving vascular flexibility, and an effect of improving blood vessel-dependent vasodilatory function, and PB3 with respect to PB1. When the ratio is 0.41 to 0.68, the vascular endothelial function improving effect, the vascular flexibility improving effect, and the blood vessel-dependent vasodilator function improving effect are more prominently exhibited, and the ratio of PB3 to PB1 is 0. When it was .41 to 0.60, it was found that the vascular endothelial function improving effect, the vascular flexibility improving effect, and the blood vessel-dependent vasodilator function improving effect were extremely remarkably exhibited.
製造例：実施例の結果に基づいて、以下の製造例を提示する。 Production Example: The following production example is presented based on the results of the examples.
（製造例１）
健康食品（錠剤）
松樹皮抽出物 ３０ｇ
（ＰＢ１とＰＢ３を合計して、５質量％含む。ＰＢ１：ＰＢ３＝１：０．６）
結晶セルロース １０ｇ
ショ糖脂肪酸エステル ５ｇ
二酸化ケイ素 ２ｇ
還元麦芽糖 ５３ｇ
を混合・撹拌して均一に調整し、打錠し、１錠２５０ｍｇである錠剤を製造した。得られた錠剤を１日２回、１回当たり２粒摂取することで、優れた血管内皮機能改善作用、血管柔軟性向上作用、及び血流依存性血管拡張機能改善作用が得られる。
(Manufacturing Example 1)
Health food (tablet)
Pine bark extract 30g
(PB1 and PB3 are totaled and include 5% by mass. PB1: PB3 = 1: 0.6)
Crystalline cellulose 10g
Sucrose fatty acid ester 5g
Silicon dioxide 2g
Reduced maltose 53g
Was mixed and stirred to make a uniform adjustment, and the tablets were beaten to produce tablets having a dose of 250 mg per tablet. By ingesting the obtained tablets twice a day, 2 tablets at a time, an excellent vascular endothelial function improving action, a vascular flexibility improving action, and a blood flow-dependent vasodilator function improving action can be obtained.
（製造例２）
健康食品（カプセル剤）
松樹皮抽出物 ３ｍｇ
（ＰＢ１とＰＢ３を合計して、８０質量％含む。ＰＢ１：ＰＢ３＝１：０．７６）
ココナッツミルク末 ６０ｍｇ
イソマルトオリゴ糖 ６０ｍｇ
麦芽糖 ６０ｍｇ
二酸化ケイ素 １０ｍｇ
ナイアシン １ｍｇ
マグネシウム １ｍｇ
を混合・撹拌して均一に調整し、ハードカプセルに封入し、カプセル剤を製造した。得られたカプセル剤を１日１回、１回当たり１粒摂取することで、優れた血管内皮機能改善作用、血管柔軟性向上作用、及び血流依存性血管拡張機能改善作用が得られる。
(Manufacturing Example 2)
Health food (capsule)
Pine bark extract 3 mg
(PB1 and PB3 are totaled and include 80% by mass. PB1: PB3 = 1: 0.76)
Coconut milk powder 60mg
Isomaltooligosaccharide 60 mg
Maltose 60mg
Silicon dioxide 10 mg
Niacin 1 mg
Magnesium 1 mg
Was mixed and stirred to make a uniform adjustment, and the mixture was encapsulated in a hard capsule to produce a capsule. By ingesting one capsule of the obtained capsule once a day, an excellent effect of improving vascular endothelial function, an effect of improving vascular flexibility, and an effect of improving blood flow-dependent vasodilator function can be obtained.
（製造例３）
健康食品（顆粒剤）
松樹皮抽出物 ５ｇ
（ＰＢ１とＰＢ３を合計して、８質量％含む。ＰＢ１：ＰＢ３＝１：１．４０）
大麦若葉末 ５０ｇ
フラクトオリゴ糖 １０ｇ
イソマルトオリゴ糖 １０ｇ
還元麦芽糖 ３０ｇ
ショ糖脂肪酸エステル ３ｇ
二酸化ケイ素 ２ｇ
を混合・撹拌して均一に調整し、顆粒剤を製造した。得られた顆粒剤２ｇを１００ｇの水に溶解し、１日１回摂取することで、優れた血管内皮機能改善作用、血管柔軟性向上作用、及び血流依存性血管拡張機能改善作用が得られる。
(Manufacturing Example 3)
Health food (granule)
Pine bark extract 5g
(PB1 and PB3 are totaled and include 8% by mass. PB1: PB3 = 1: 1.40)
Barley young leaf powder 50g
Fructooligosaccharide 10g
Isomaltooligosaccharide 10g
Reduced maltose 30g
Sucrose fatty acid ester 3g
Silicon dioxide 2g
Was mixed and stirred to make a uniform adjustment to produce granules. By dissolving 2 g of the obtained granules in 100 g of water and ingesting it once a day, excellent vascular endothelial function improving action, vascular flexibility improving action, and blood flow-dependent vasodilator function improving action can be obtained. ..
ＰＢ１及びＰＢ３を含有すことを特徴とする組成物に優れた血流依存性血管拡張機能の改善が見出されたことから本発明は、優れた血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物を提供することができる。優れた血管内皮機能改善用組成物、血管柔軟性向上用組成物、及び血流依存性血管拡張機能改善用組成物は、血管内皮機能改善作用、血管柔軟性向上作用、及び血流依存性血管拡張機能改善作用を有し、経口剤として用いることができることから、本発明の産業上の有用性は高い。 Since an excellent improvement in blood flow-dependent vasodilatory function was found in a composition characterized by containing PB1 and PB3, the present invention is an excellent composition for improving vascular endothelial function and improving vascular flexibility. And a composition for improving blood flow-dependent vasodilatory function can be provided. The excellent composition for improving vascular endothelial function, the composition for improving vascular flexibility, and the composition for improving blood flow-dependent vasodilatory function have an effect of improving vascular endothelial function, an effect of improving vascular flexibility, and a blood flow-dependent blood vessel. The present invention is highly industrially useful because it has an effect of improving diastolic function and can be used as an oral preparation.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4698360A (en) * | 1985-04-09 | 1987-10-06 | Societe Civile D'investigations Pharmacologiques D'aquitaine | Plant extract with a proanthocyanidins content as therapeutic agent having radical scavenger effect and use thereof |
| JP2001106634A (en) * | 1999-10-08 | 2001-04-17 | Nobutaka Suzuki | Therapeutic drug for dysmenorrhea or endometriosis |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4698360A (en) * | 1985-04-09 | 1987-10-06 | Societe Civile D'investigations Pharmacologiques D'aquitaine | Plant extract with a proanthocyanidins content as therapeutic agent having radical scavenger effect and use thereof |
| US4698360B1 (en) * | 1985-04-09 | 1997-11-04 | D Investigations Pharmacologiq | Plant extract with a proanthocyanidins content as therapeutic agent having radical scavenger effect and use thereof |
| JP2001106634A (en) * | 1999-10-08 | 2001-04-17 | Nobutaka Suzuki | Therapeutic drug for dysmenorrhea or endometriosis |
Non-Patent Citations (4)
| Title |
|---|
| "スギおよびアカマツ樹皮中でのフラバノール類の蓄積ならびにその構造上の変化", 東京大学農学部演習林報告, vol. 72, JPN6021032669, 1982, pages 17 - 29, ISSN: 0004692113 * |
| JOURNAL OF CARDIOVASCULAR PHARMACOLOGY, vol. 32, JPN6021022631, 1998, pages 509 - 515, ISSN: 0004692111 * |
| JPN PHARMACOL THER, vol. 48, no. 5, JPN6021032668, 20 May 2020 (2020-05-20), pages 853 - 858, ISSN: 0004692114 * |
| NEW FOOD INDUSTRY, vol. 46, no. 5, JPN6021022634, 2004, pages 1 - 7, ISSN: 0004692112 * |
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