JP2003300996A - New angiotensin i converting enzyme inhibitor - Google Patents
New angiotensin i converting enzyme inhibitorInfo
- Publication number
- JP2003300996A JP2003300996A JP2002110456A JP2002110456A JP2003300996A JP 2003300996 A JP2003300996 A JP 2003300996A JP 2002110456 A JP2002110456 A JP 2002110456A JP 2002110456 A JP2002110456 A JP 2002110456A JP 2003300996 A JP2003300996 A JP 2003300996A
- Authority
- JP
- Japan
- Prior art keywords
- peptide
- ace
- angiotensin
- converting enzyme
- active ingredient
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Links
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Peptides Or Proteins (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、アンジオテンシン
Ｉ変換酵素（以下、「ＡＣＥ」ともいう）を阻害する作
用を有する新規なＡＣＥ阻害剤、及び該ＡＣＥ阻害剤を
有効成分として含有するＡＣＥ阻害作用を有する医薬用
及び食品用組成物に関する。TECHNICAL FIELD The present invention relates to a novel ACE inhibitor having an action of inhibiting angiotensin I converting enzyme (hereinafter, also referred to as “ACE”), and an ACE inhibitory action containing the ACE inhibitor as an active ingredient. The present invention relates to a pharmaceutical composition and a food composition.
【０００２】[0002]
【従来の技術】高血圧症は、循環器系の種々の疾病の重
要な危険因子であり、血圧を正常な範囲に保つことは、
健康の維持に重要であるが、平成１０年の国民栄養調査
によれば、１５歳以上の男性１９．８％、女性１４．５
％が高血圧である。BACKGROUND ART Hypertension is an important risk factor for various diseases of the circulatory system, and keeping blood pressure in a normal range is
It is important for maintaining good health, but according to the 1998 National Nutrition Survey, 19.8% for men over 15 years old, 14.5% for women
% Have high blood pressure.
【０００３】高血圧症の多くは、明確にその原因が特定
できない本態性高血圧症であるが、近年、遺伝子レベル
で高血圧症とレニン・アンジオテンシン系との関連を示
唆する報告がでる等、高血圧症の要因の一つとしてＡＣ
Ｅの作用異常が挙げられるようになった。Most of the hypertensions are essential hypertensions whose cause cannot be clearly identified. In recent years, however, it has been reported that hypertension is associated with the renin-angiotensin system at the gene level. AC as one of the factors
The abnormal action of E came to be mentioned.
【０００４】ＡＣＥは、主に２つの血圧調節系に関与し
て昇圧作用を示す酵素であるが、この酵素が異常に作用
すると高血圧症を誘発するとされる。そこで、このよう
なＡＣＥの昇圧作用を阻害する物質を投与することによ
り高血圧症を改善しようとする試みがなされ、目立った
副作用もないことから全高血圧治療薬に占める割合も増
加している。[0004] ACE is an enzyme mainly involved in two blood pressure control systems and showing a pressor action, and it is said that hyperactivity is induced when this enzyme acts abnormally. Therefore, attempts have been made to improve hypertension by administering such a substance that inhibits the pressor action of ACE, and since there are no conspicuous side effects, the proportion of all antihypertensive drugs is increasing.
【０００５】一方、食品の三次機能としての生体調節機
能の解明が様々な食品についてなされ、その過程におい
て多くのＡＣＥ阻害物質を単離した報告も数多くなされ
ている。また、このような食品等から得られたＡＣＥ阻
害物質の中には、高血圧症のモデル動物への投与で効果
が確認されているものもあり、このような特徴を利用し
た機能性食品の開発もなされてきている。On the other hand, elucidation of the bioregulatory function as the tertiary function of foods has been made for various foods, and there have been many reports that many ACE inhibitors have been isolated in the process. In addition, some ACE inhibitors obtained from such foods have been confirmed to be effective when administered to model animals with hypertension, and development of functional foods utilizing such characteristics Has also been done.
【０００６】ところで、光合成を行う緑色植物の葉緑素
には、ルビスコ（RuBisCO: ribulose-1,5-bisphosphate
carboxylase/ oxygenase）と呼ばれる酵素が含まれて
いる。本酵素は光合成の暗反応において、リブロース−
１，５−二リン酸と二酸化炭素から二分子のグリセリン
酸−３−リン酸を生成することにより、二酸化炭素をカ
ルビン回路に取り込む働きを持っており、地球上におい
てもっとも量が多い酵素であるとも言われる。現在まで
に、ルビスコからＡＣＥ阻害剤を分離した又はルビスコ
からＡＣＥ阻害ペプチドを単離し、その構造を決定した
という報告は知られていない。By the way, rubisco (RuBisCO: ribulose-1,5-bisphosphate) is known as chlorophyll for photosynthetic green plants.
It contains an enzyme called carboxylase / oxygenase). In the dark reaction of photosynthesis, this enzyme is ribulose-
By producing two molecules of glyceric acid-3-phosphoric acid from 1,5-diphosphoric acid and carbon dioxide, it has a function of taking carbon dioxide into the Calvin cycle and is the most abundant enzyme on earth. It is also said. To date, there are no reports of separating the ACE inhibitor from Rubisco or isolating the ACE inhibitory peptide from Rubisco and determining its structure.
【０００７】[0007]
【発明が解決しようとする課題】本発明は上記観点から
なされたものであり、ＡＣＥ阻害作用を有する新規なＡ
ＣＥ阻害剤、及び該ＡＣＥ阻害剤を有効成分として含有
する医薬用組成物又は食品用組成物を提供することを課
題とする。SUMMARY OF THE INVENTION The present invention has been made from the above viewpoint, and is a novel A having an ACE inhibitory action.
It is an object to provide a CE inhibitor and a pharmaceutical composition or food composition containing the ACE inhibitor as an active ingredient.
【０００８】[0008]
【課題を解決するための手段】本発明者らは、上記課題
を解決するため鋭意研究を重ねた結果、ルビスコのペプ
シン又はペプシンとパンクレアチンによる加水分解物、
及び該加水分解物から得た画分にＡＣＥ阻害作用がある
ことを見出した。さらに、該画分を単離、構造決定した
ところ、新規なＡＣＥ阻害ペプチドであることを同定
し、本発明を完成するに至った。Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have found that rubisco pepsin or a hydrolyzate of pepsin and pancreatin,
And it was found that the fraction obtained from the hydrolyzate had an ACE inhibitory action. Furthermore, when the fraction was isolated and the structure was determined, it was identified as a novel ACE-inhibiting peptide, and the present invention was completed.
【０００９】すなわち、本発明は、以下に示すものであ
る。
（１）配列番号１のアミノ酸配列を有し、かつアンジオ
テンシンＩ変換酵素を阻害する作用を有するペプチド。
（２）配列番号１において、アミノ酸番号１〜３で表さ
れる配列を有する、（１）に記載のペプチド。
（３）配列番号２のアミノ酸配列を有する、（１）に記
載のペプチド。
（４）（１）〜（３）の何れかに記載のペプチドを有効
成分として含有するアンジオテンシンＩ変換酵素阻害
剤。
（５）リブロース−１，５−ビスリン酸カルボキシラー
ゼ／オキシゲナーゼをペプシンで又はペプシンとパンク
レアチンで加水分解して得られる加水分解物を有効成分
として含有する、（４）に記載のアンジオテンシンＩ変
換酵素阻害剤。
（６）（４）に記載のペプチド及び／又はその生理的に
許容される塩の１種又は２種以上、及び／又は、（５）
に記載の加水分解物及び／又はその生理的に許容される
塩の１種又は２種以上を有効成分として含有するアンジ
オテンシンＩ変換酵素阻害剤を、有効成分として含有す
る医薬用組成物。
（７）（４）に記載のペプチド及び／又はその生理的に
許容される塩の１種又は２種以上、及び／又は、（５）
に記載の加水分解物及び／又はその生理的に許容される
塩の１種又は２種以上を有効成分として含有するアンジ
オテンシンＩ変換酵素阻害剤を、有効成分として含有す
る食品用組成物。
（８）リブロース−１，５−ビスリン酸カルボキシラー
ゼ／オキシゲナーゼをペプシンで又はペプシンとパンク
レアチンで加水分解して得られる加水分解物からアンジ
オテンシンＩ変換酵素を阻害する作用を有するペプチド
を採取することを特徴とするアンジオテンシンＩ変換酵
素を阻害する作用を有するペプチドの製造方法。
（９）前記アンジオテンシンＩ変換酵素を阻害する作用
を有するペプチドが（１）〜（３）の何れかに記載のペ
プチドである、（８）に記載のペプチドの製造方法。That is, the present invention is as follows. (1) A peptide having the amino acid sequence of SEQ ID NO: 1 and having an action of inhibiting angiotensin I converting enzyme. (2) The peptide according to (1), which has a sequence represented by amino acid numbers 1 to 3 in SEQ ID NO: 1. (3) The peptide according to (1), which has the amino acid sequence of SEQ ID NO: 2. (4) An angiotensin I converting enzyme inhibitor containing the peptide according to any one of (1) to (3) as an active ingredient. (5) Angiotensin I-converting enzyme inhibitor according to (4), which contains a hydrolyzate obtained by hydrolyzing ribulose-1,5-bisphosphate carboxylase / oxygenase with pepsin or with pepsin and pancreatin as an active ingredient. Agent. (6) One or more kinds of the peptide according to (4) and / or a physiologically acceptable salt thereof, and / or (5)
A pharmaceutical composition comprising, as an active ingredient, an angiotensin I-converting enzyme inhibitor containing, as an active ingredient, one or more of the hydrolyzate and / or a physiologically acceptable salt thereof described in (1). (7) One or more kinds of the peptide according to (4) and / or a physiologically acceptable salt thereof, and / or (5)
A food composition comprising as an active ingredient an angiotensin I-converting enzyme inhibitor containing one or two or more of the hydrolyzate and / or physiologically acceptable salt thereof according to claim 1 as an active ingredient. (8) A peptide having an action of inhibiting angiotensin I converting enzyme is collected from a hydrolyzate obtained by hydrolyzing ribulose-1,5-bisphosphate carboxylase / oxygenase with pepsin or with pepsin and pancreatin. And a method for producing a peptide having an action of inhibiting angiotensin I converting enzyme. (9) The method for producing a peptide according to (8), wherein the peptide having an action of inhibiting the angiotensin I converting enzyme is the peptide according to any one of (1) to (3).
【００１０】[0010]
【発明の実施の形態】以下、本発明を詳細に説明する。
＜１＞本発明のアンジオテンシンＩ変換酵素阻害剤
本発明のＡＣＥ阻害剤は、新規な化合物である本発明の
ＡＣＥ阻害ペプチドを有効成分として含有する。BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below. <1> Angiotensin I-Converting Enzyme Inhibitor of the Present Invention The ACE inhibitor of the present invention contains the ACE inhibitory peptide of the present invention, which is a novel compound, as an active ingredient.
【００１１】本発明のＡＣＥ阻害ペプチドは、Ｍｅｔ−
Ａｒｇ−Ｔｒｐ−Ｘａａ−Ｘａａ（配列番号１）のアミ
ノ酸配列を有し、かつＡＣＥ阻害作用を有するペプチド
である。The ACE-inhibiting peptide of the present invention is Met-
It is a peptide having an amino acid sequence of Arg-Trp-Xaa-Xaa (SEQ ID NO: 1) and having an ACE inhibitory action.
【００１２】ここで、配列番号１におけるＸａａは、そ
れぞれ独立に任意のアミノ酸、又は存在しないことを表
す。Here, Xaa in SEQ ID NO: 1 independently represents any amino acid or does not exist.
【００１３】上記ペプチドとして具体的には、Ｍｅｔ−
Ａｒｇ−Ｔｒｐのアミノ酸配列（配列番号１において、
アミノ酸番号１〜３で表される配列）を有するペプチド
が挙げられる。The above-mentioned peptide is specifically Met-
The amino acid sequence of Arg-Trp (in SEQ ID NO: 1,
Peptides having amino acid numbers 1 to 3) are included.
【００１４】また、Ｍｅｔ−Ａｒｇ−Ｔｒｐ−Ａｒｇ−
Ａｓｐのアミノ酸配列（配列番号２）を有するペプチド
が挙げられる。Further, Met-Arg-Trp-Arg-
A peptide having the amino acid sequence of Asp (SEQ ID NO: 2) can be mentioned.
【００１５】これら本発明のＡＣＥ阻害ペプチドは、ア
ミノ酸配列が明らかなため化学的に合成することにより
得ることもできるし、例えば下記に示す方法等により、
ルビスコをペプシン又はペプシンとパンクレアチンで加
水分解して得られる加水分解物から、単離することによ
り得ることもできる。These ACE-inhibiting peptides of the present invention can be obtained by chemical synthesis because the amino acid sequence is clear. For example, by the method shown below,
It can also be obtained by isolating from the hydrolyzate obtained by hydrolyzing rubisco with pepsin or pepsin and pancreatin.
【００１６】本発明のＡＣＥ阻害ペプチドの高速液体ク
ロマトグラフィー（以下、「ＨＰＬＣ」ともいう）を用
いた抽出例を以下に例示するが、本発明はこの抽出例に
限定されるものではない。
（１）ルビスコの加水分解物の調製
ルビスコの溶解液にペプシンを加えて反応させる。酵素
反応を終了させた後、遠心分離により不溶物を除き、得
られた上清を凍結乾燥してペプシンの加水分解物を得
る。The extraction example of the ACE-inhibiting peptide of the present invention using high performance liquid chromatography (hereinafter, also referred to as "HPLC") is illustrated below, but the present invention is not limited to this extraction example. (1) Preparation of hydrolyzate of rubisco Pepsin is added to a solution of rubisco to react. After the enzymatic reaction is completed, the insoluble matter is removed by centrifugation, and the obtained supernatant is lyophilized to obtain a hydrolyzate of pepsin.
【００１７】また、上記操作において、ペプシンの反応
後にパンクレアチンを加えて反応させ、該反応液に対し
酵素反応を終了させた後、遠心分離で不溶物を除き、得
られた上清を凍結乾燥すれば、ペプシン−パンクレアチ
ンの加水分解物が得られる。
（２）ＨＰＬＣによる分画
上述した加水分解物に対し、溶出液の組成を適宜変更さ
せた条件で、ＨＰＬＣを用いてクロマトグラフ分画を行
う。目的となるペプチドの活性は、以下に記載の実施例
の方法で測定できる。In the above operation, after the reaction of pepsin, pancreatin is added and reacted, the enzymatic reaction is terminated to the reaction solution, the insoluble matter is removed by centrifugation, and the obtained supernatant is freeze-dried. Then, a hydrolyzate of pepsin-pancreatin is obtained. (2) Fractionation by HPLC Chromatographic fractionation is performed on the above-mentioned hydrolyzate using HPLC under the condition that the composition of the eluate is appropriately changed. The activity of the peptide of interest can be measured by the method of Examples described below.
【００１８】それにより、配列番号１においてアミノ酸
番号１〜３で表されるアミノ酸配列を有するＡＣＥ阻害
ペプチド又は配列番号２のアミノ酸配列を有するＡＣＥ
阻害ペプチドの分画物を単離することができる。Thereby, the ACE-inhibiting peptide having the amino acid sequence represented by amino acid numbers 1 to 3 in SEQ ID NO: 1 or the ACE having the amino acid sequence of SEQ ID NO: 2
Fractions of inhibitory peptides can be isolated.
【００１９】分画により単離した化合物の構造決定は、
以下実施例でも示しているように、ペプチドシーケンサ
ーにより定法に従って行った。The structure of the compound isolated by fractionation was determined by
As shown in the following examples, the procedure was carried out according to a standard method using a peptide sequencer.
【００２０】上記の様にしてルビスコの加水分解物から
単離された本発明のＡＣＥ阻害ペプチドは、以下の実施
例で示すようにＡＣＥ阻害作用を有するため、ＡＣＥ阻
害剤として有効に使用することができる。尚、配列番号
２のアミノ酸配列を有する阻害ペプチドは、以下実施例
で述べているようにプロドラッグタイプである可能性が
あるが、いずれにしても、配列番号１においてアミノ酸
番号１〜３で表されるアミノ酸配列を有するＡＣＥ阻害
ペプチド及び配列番号２のアミノ酸配列を有するＡＣＥ
阻害ペプチドの両者とも本発明でいうＡＣＥ阻害効果が
期待できるものとなっている。The ACE-inhibiting peptide of the present invention isolated from the hydrolyzate of rubisco as described above has an ACE-inhibiting action as shown in the following examples, and therefore should be effectively used as an ACE inhibitor. You can The inhibitory peptide having the amino acid sequence of SEQ ID NO: 2 may be a prodrug type as described in Examples below, but in any case, it is represented by amino acid numbers 1 to 3 in SEQ ID NO: 1. ACE-inhibiting peptide having the amino acid sequence shown in FIG.
Both of the inhibitory peptides can be expected to have the ACE inhibitory effect according to the present invention.
【００２１】また上記において、ペプチドを単離する前
のルビスコの加水分解物又はその粗精製物も、該加水分
解物中に含まれる本発明のＡＣＥ阻害ペプチドの示すＡ
ＣＥ阻害作用を利用することができ、以下の実施例で示
すようにＡＣＥ阻害作用を示す。Further, in the above, the hydrolyzate of rubisco or a crudely purified product thereof prior to the isolation of the peptide is the A represented by the ACE-inhibiting peptide of the present invention contained in the hydrolyzate.
The CE inhibitory action can be utilized, and it exhibits the ACE inhibitory action as shown in the following examples.
【００２２】よって、ルビスコの加水分解物を有効成分
として含有する剤も、本発明のＡＣＥ阻害剤として有効
に使用することができる。Therefore, an agent containing a hydrolyzate of rubisco as an active ingredient can also be effectively used as the ACE inhibitor of the present invention.
【００２３】このように、本発明のＡＣＥ阻害ペプチド
又はルビスコの加水分解物、又はそれらの生理的に許容
される塩は、１種又は２種以上組み合わせて、本発明の
効果を発揮するに有効な量含有させて、ＡＣＥ阻害剤と
して利用することができる。As described above, the ACE-inhibiting peptide of the present invention, the hydrolyzate of rubisco, or a physiologically acceptable salt thereof is effective in exerting the effects of the present invention by combining one or two or more thereof. It can be used as an ACE inhibitor by containing a large amount.
【００２４】ここで、上記「生理的に許容される塩」と
は、生理的に許容されるものであれば特段の限定はされ
ないが、例えば、ナトリウム塩やカリウム塩等のアルカ
リ金属塩、カルシウム塩やマグネシウム塩などのアルカ
リ土類金属塩、アンモニウム塩、トリエチルアミン塩や
トリエタノールアミン塩等の有機アミン塩、又はリジン
塩やアルギニン塩等の塩基性アミノ酸塩が好適に例示で
きる。Here, the "physiologically acceptable salt" is not particularly limited as long as it is physiologically acceptable. For example, alkali metal salts such as sodium salt and potassium salt, calcium, etc. Preferable examples thereof include alkaline earth metal salts such as salts and magnesium salts, ammonium salts, organic amine salts such as triethylamine salts and triethanolamine salts, and basic amino acid salts such as lysine salts and arginine salts.
【００２５】またさらに上記ＡＣＥ阻害剤と、医薬用又
は食品用として通常用いられている他の任意成分とを組
み合わせれば、ＡＣＥ阻害作用を有する医薬用組成物や
食品用組成物を提供することができる。Further, by combining the above-mentioned ACE inhibitor with other optional components usually used for medicine or food, it is possible to provide a pharmaceutical composition or food composition having an ACE inhibitory action. You can
【００２６】尚、本発明において、ＡＣＥ阻害作用を示
すか否かを判断する方法としては、後記実施例に示す通
り、公知の測定方法を利用した評価方法を用いることが
できる。
＜２＞本発明のアンジオテンシンＩ変換酵素阻害剤を含
有する医薬用組成物
本発明の医薬用組成物は、上記本発明のＡＣＥ阻害ペプ
チド及び／又はその生理的に許容される塩の１種又は２
種以上、及び／又は、上記のルビスコの加水分解物及び
／又はその生理的に許容される塩の１種又は２種以上を
有効成分として含有する本発明のＡＣＥ阻害剤を、有効
成分として含有する。In the present invention, as a method for determining whether or not it exhibits an ACE inhibitory action, an evaluation method utilizing a known measuring method can be used, as shown in Examples below. <2> Pharmaceutical Composition Containing Angiotensin I-Converting Enzyme Inhibitor of the Present Invention The pharmaceutical composition of the present invention is one of the above-mentioned ACE-inhibiting peptide of the present invention and / or a physiologically acceptable salt thereof, or Two
Containing, as an active ingredient, an ACE inhibitor of the present invention containing one or more hydrolyzate of rubisco and / or one or more kinds of physiologically acceptable salts thereof as an active ingredient. To do.
【００２７】本発明の医薬用組成物は、本発明のＡＣＥ
阻害剤を、定法に従って配合したものであり、ＡＣＥ阻
害作用が期待できるものであれば特に限定されるもので
はない。The pharmaceutical composition of the present invention is the ACE of the present invention.
The inhibitor is compounded according to a standard method, and is not particularly limited as long as an ACE inhibitory effect can be expected.
【００２８】本発明の医薬用組成物の剤型は、特に限定
されないが、一般に製剤上許容される１または２種類以
上の担体、賦型剤、統合剤、防腐剤、安定剤、香味剤等
と共に混合して、錠剤、顆粒剤、カプセル剤、水薬、ド
リンク剤等の内服剤型とすることが好ましい。このよう
な製剤化は、通常、医薬品の製造に用いられる方法に従
って製剤化することができる。The dosage form of the pharmaceutical composition of the present invention is not particularly limited, but generally one or more kinds of carriers, excipients, integrators, preservatives, stabilizers, flavoring agents and the like which are generally acceptable in the formulation. It is preferable to mix them together with each other to give an oral dosage form such as tablets, granules, capsules, drenches, and drinks. Such a formulation can be prepared according to a method usually used for manufacturing a pharmaceutical product.
【００２９】上記医薬用組成物の投与量としては、症
状、患者の年齢、体重等により異なるが、成人１日当た
り、上記本発明のＡＣＥ阻害ペプチドを０．１〜５０mg
含む、又は上記ルビスコの加水分解物を０．２〜５０g
含むＡＣＥ阻害剤を含有する医薬用組成物を、１回ない
し数回に分けて経口投与するのが好ましい。
＜３＞本発明のアンジオテンシンＩ変換酵素阻害剤を含
有する食品用組成物
本発明の食品用組成物は、上記本発明のＡＣＥ阻害ペプ
チド及び／又はその生理的に許容される塩の１種又は２
種以上、及び／又は、上記のルビスコの加水分解物及び
／又はその生理的に許容される塩の１種又は２種以上を
有効成分として含有する本発明のＡＣＥ阻害剤を、有効
成分として含有する。The dose of the above-mentioned pharmaceutical composition varies depending on the symptoms, age and weight of the patient, but 0.1-50 mg of the above-mentioned ACE inhibitory peptide of the present invention per adult per day
0.2 to 50 g of the hydrolyzate of Rubisco
The pharmaceutical composition containing the ACE inhibitor is preferably administered orally in one to several divided doses. <3> Composition for Food Containing Angiotensin I-Converting Enzyme Inhibitor of the Present Invention The food composition of the present invention is one of the above-mentioned ACE-inhibiting peptide of the present invention and / or a physiologically acceptable salt thereof. Two
Containing, as an active ingredient, an ACE inhibitor of the present invention containing one or more hydrolyzate of rubisco and / or one or more kinds of physiologically acceptable salts thereof as an active ingredient. To do.
【００３０】本発明の食品用組成物は、本発明のＡＣＥ
阻害剤を、常法にしたがって配合したものである。The food composition of the present invention is the ACE of the present invention.
The inhibitor is compounded according to a conventional method.
【００３１】本発明の食品用組成物としては、ＡＣＥ阻
害作用が期待できるものであれば特に限定されるもので
はないが、種々の食品に、食品として通常用いられてい
る任意成分とともに、食品原料に抽出物を所要量配合す
ることができる。この抽出物を配合する際に特に留意す
ることはなく、通常の製造方法により加工製造すること
により、健康食品、機能性食品を製造することができ
る。配合量は、食品の種類により異なるが、食品の味を
損なわず、且つ十分なＡＣＥ阻害効果を得るためには、
食品用組成物全量に対して、本発明のＡＣＥ阻害ペプチ
ドを０．００１〜１（重量％）の割合で、又はルビスコ
の加水分解物を１〜５０（重量％）の割合で含むＡＣＥ
阻害剤を、配合させるのが望ましい。The food composition of the present invention is not particularly limited as long as it can be expected to have an ACE inhibitory action, but it is used in various foods together with optional ingredients usually used as foods and food raw materials. The extract can be mixed with the required amount. No particular attention is paid when blending this extract, and health foods and functional foods can be manufactured by processing and manufacturing by ordinary manufacturing methods. The blending amount varies depending on the type of food, but in order to obtain a sufficient ACE inhibitory effect without impairing the taste of the food,
ACE containing 0.001 to 1 (wt%) of the ACE-inhibiting peptide of the present invention or 1 to 50 (wt%) of hydrolyzate of rubisco with respect to the total amount of the food composition.
It is desirable to incorporate an inhibitor.
【００３２】[0032]
【実施例】以下、本発明を実施例によりさらに具体的に
説明する。EXAMPLES The present invention will be described in more detail below with reference to examples.
【００３３】[0033]
【実施例１】＜ルビスコ加水分解物の調製及びＡＣＥ阻
害ペプチドの単離＞
＜１＞ルビスコ加水分解物の調製
（１）ペプシンによる加水分解物の調製
ホウレンソウ由来のルビスコ（Sigma 社製、100mg）を1
0 mLの純水に懸濁し、塩酸を加えてpH 2.0で溶解し、ペ
プシン（Sigma 社製）1 mgを添加して37℃にて５時間反
応させた。Example 1 <Preparation of Rubisco Hydrolyzate and Isolation of ACE Inhibitory Peptide><1> Preparation of Rubisco Hydrolyzate (1) Preparation of Hydrolyzate with Pepsin Rubisco derived from spinach (Sigma, 100 mg) 1
It was suspended in 0 mL of pure water, hydrochloric acid was added to dissolve it at pH 2.0, 1 mg of pepsin (manufactured by Sigma) was added, and the mixture was reacted at 37 ° C for 5 hours.
【００３４】続いて、1M NaOHを加えてpH7.5に調整して
酵素反応を終了させ、さらに遠心分離（10,000 rpm、10
min）により不溶物を除き、得られた上清を凍結乾燥し
てペプシン加水分解物を得た。Subsequently, 1 M NaOH was added to adjust the pH to 7.5 to terminate the enzyme reaction, and further centrifugation (10,000 rpm, 10
min) to remove insoluble matter, and the resulting supernatant was lyophilized to obtain a pepsin hydrolyzate.
【００３５】（２）ペプシン−パンクレアチンによる加
水分解物の調製
ホウレンソウ由来のルビスコ（Sigma 社製、100mg）を1
0 mLの純水に懸濁し、塩酸を加えてpH 2.0で溶解し、ペ
プシン（Sigma 社製）1 mgを添加して37℃にて５時間反
応させた。続いて、1M NaOHを加えてpH7.5に調整した
後、パンクレアチン（Sigma 社製）を5 mg添加し、37℃
にて５時間酵素処理した。(2) Preparation of Hydrolyzate with Pepsin-Pancreatine 1 part of Rubisco (Sigma, 100 mg) derived from spinach was used.
It was suspended in 0 mL of pure water, hydrochloric acid was added to dissolve it at pH 2.0, 1 mg of pepsin (manufactured by Sigma) was added, and the mixture was reacted at 37 ° C for 5 hours. Next, adjust the pH to 7.5 by adding 1M NaOH, and then add 5 mg of pancreatin (Sigma) and incubate at 37 ℃.
Was treated with an enzyme for 5 hours.
【００３６】続いて、塩酸を加えてpH2.0に調整して酵
素反応を終了させ、さらに遠心分離（10,000 rpm、10 m
in）により不溶物を除き、得られた上清を凍結乾燥し、
ペプシン−パンクレアチン加水分解物を得た。
＜２＞ＨＰＬＣによるペプチドの単離及び構造決定
上記実施例１の＜１＞（２）で得られたペプシン−パン
クレアチン加水分解物を純水に再溶解し、以下表１で示
す条件で逆相カラムを順番に用い、ＨＰＬＣにより、Ａ
ＣＥ阻害ペプチドを単離した。Subsequently, hydrochloric acid was added to adjust the pH to 2.0 to terminate the enzymatic reaction, and further centrifugation (10,000 rpm, 10 m
in) to remove insoluble matter and freeze-dry the resulting supernatant,
A pepsin-pancreatin hydrolyzate was obtained. <2> Isolation and Structure Determination of Peptide by HPLC The pepsin-pancreatin hydrolyzate obtained in <1> (2) of Example 1 above was redissolved in pure water and reversed under the conditions shown in Table 1 below. By using a phase column in sequence, by HPLC, A
The CE inhibitory peptide was isolated.
【００３７】[0037]
【表１】
表１ ＡＣＥ阻害物質の単離に用いたＨＰＬＣの条件
──────────────────────────────────
カラム 流速（mL/min）
──────────────────────────────────
ＯＤＳ １０．０
ＰｈＡ １．０
ＣＮ １．０
Ｎ−ＰｈＡ １．０
──────────────────────────────────
溶媒：アセトニトリル(0.1％トリフルオロ酢酸)／純水(0.1％トリフルオロ酢酸)
濃度勾配：アセトニトリル(0.1％トリフルオロ酢酸)：０→４０％ for 40分
カラム：ＯＤＳ・・・Cosmosil 5C18-ARII、Ф20mm×250mm(ナカライテスク社製)
ＰｈＡ・・・Cosmosil 5PE-MS、Ф4.6mm×250mm(ナカライテスク社製)
ＣＮ・・・Cosmosil 5CN-R、Ф4.6mm×250mm(ナカライテスク社製)
Ｎ−ＰｈＡ・・・Cosmosil 5NPE、Ф4.6mm×150mm(ナカライテスク社製)
（１）ＯＤＳカラムを用いたＨＰＬＣによる分画を行っ
た（図１）。図１中、Ｉ〜IIIの３箇所にＡＣＥ阻害活
性が認められた。[Table 1] Table 1 Conditions of HPLC used for isolation of ACE inhibitor ───────────────────────────────── ── Column Flow rate (mL / min) ────────────────────────────────── ODS 10.0 PhA 1. 0 CN 1.0 N-PhA 1.0 ────────────────────────────────── Solvent: Acetonitrile (0.1 % Trifluoroacetic acid) / Pure water (0.1% trifluoroacetic acid) Concentration gradient: Acetonitrile (0.1% trifluoroacetic acid): 0 → 40% for 40 minutes Column: ODS ・ ・ ・ Cosmosil 5C 18 -ARII, φ20mm × 250mm ( PhA ・ ・ ・ Cosmosil 5PE-MS, Φ4.6mm × 250mm (manufactured by Nacalai Tesque) CN ・ ・ ・ Cosmosil 5CN-R, Φ4.6mm × 250mm (manufactured by Nacalai Tesque) N-PhA ・ ・・ Cosmosil 5NPE, Φ 4.6 mm × 150 mm (manufactured by Nacalai Tesque, Inc.) (1) Fractionation by HPLC using an ODS column was performed (FIG. 1). In FIG. 1, ACE inhibitory activity was observed at three points I to III.
【００３８】尚、図１中、ＯＤＳカラムによるＡＣＥ阻
害物質の分画において、画分Ｉのリテンションタイム
は、２２分、画分IIのリテンションタイムは、２７分、
画分IIIのリテンションタイムは、３１分であった。In the fractionation of the ACE inhibitor by the ODS column in FIG. 1, the retention time of fraction I was 22 minutes, the retention time of fraction II was 27 minutes,
The retention time of Fraction III was 31 minutes.
【００３９】（２）上記実施例１＜２＞（１）で得られ
た、３つの画分を、更にＰｈＡカラム、ＣＮカラム、Ｎ
−ＰｈＡカラムで順次分画し、活性画分の単離を行っ
た。フラクションIIの分離過程を図２（ａ）〜（ｃ）に
示す。(2) The three fractions obtained in the above Example 1 <2> (1) were further subjected to PhA column, CN column, N
-The fractions were sequentially fractionated on the PhA column to isolate the active fraction. The separation process of Fraction II is shown in FIGS. 2 (a) to 2 (c).
【００４０】図２（ａ）中、ＰｈＡカラムによる分画に
おいて矢印で示す画分を次のＣＮカラムに供試した。図
２（ｂ）中、ＣＮカラムによる分画において矢印で示す
画分を次のＮ−ＰｈＡカラムに供試した。In FIG. 2 (a), the fraction indicated by the arrow in the PhA column fractionation was applied to the next CN column. In FIG. 2 (b), the fraction indicated by the arrow in the fractionation by the CN column was applied to the next N-PhA column.
【００４１】尚、図２（ａ）中、矢印の画分のリテンシ
ョンタイムは、１．２分、図２（ｂ）中、矢印の画分の
リテンションタイムは、４．９分、図２（ｃ）中、矢印
の画分のリテンションタイムは、３８分であった。In FIG. 2A, the retention time of the arrowed portion is 1.2 minutes, and in FIG. 2B, the retention time of the arrowed portion is 4.9 minutes. In c), the retention time of the arrowed portion was 38 minutes.
【００４２】（３）上記実施例１＜２＞（１）で示した
ようにＯＤＳカラムで分画した画分IIを、上記実施例１
＜２＞（２）で示したようにＰｈＡカラム、ＣＮカラ
ム、Ｎ−ＰｈＡカラムで順次分画し、最終的に得られた
活性画分を、プロテインシーケンサー（アプライドバイ
オシステム社製、４９２型）にて分析し、ペプチドのア
ミノ酸配列を決定した。その結果、「Ｍｅｔ−Ａｒｇ−
Ｔｒｐ−Ａｒｇ−Ａｓｐ（以下、ＭＲＷＲＤとも略
す）」（配列番号２）で示されるアミノ酸配列を有する
ことが確認された。(3) Example 1 <2> The fraction II fractionated by the ODS column as described in (1) above was used in Example 1 above.
<2> As shown in (2), a PhA column, a CN column, and an N-PhA column are sequentially fractionated, and the finally obtained active fraction is a protein sequencer (Applied Biosystems, 492 type). And the amino acid sequence of the peptide was determined. As a result, "Met-Arg-
It was confirmed to have the amino acid sequence shown by "Trp-Arg-Asp (hereinafter, also abbreviated as MRWRD)" (SEQ ID NO: 2).
【００４３】また、上記実施例１＜２＞（１）で示した
ＯＤＳカラムで分画したIIIを、上記実施例１＜２＞
（２）で記載したのと同様の方法を用いて、ＰｈＡカラ
ム、ＣＮカラム、Ｎ−ＰｈＡカラムで順次分画し、最終
的に得られた活性画分を、プロテインシーケンサーにて
分析した。その結果、「Ｍｅｔ−Ａｒｇ−Ｔｒｐ（以
下、ＭＲＷとも略す）」（配列番号１においてアミノ酸
番号１〜３で表される配列）で示されるアミノ酸配列を
有することが確認された。Further, III fractionated by the ODS column shown in the above Example 1 <2> (1) was treated with the above Example 1 <2>.
Using the same method as described in (2), fractionation was sequentially performed on a PhA column, a CN column, and an N-PhA column, and the finally obtained active fraction was analyzed by a protein sequencer. As a result, it was confirmed to have an amino acid sequence represented by “Met-Arg-Trp (hereinafter, also abbreviated as MRW)” (sequence represented by amino acid numbers 1 to 3 in SEQ ID NO: 1).
【００４４】[0044]
【実施例２】＜ＡＣＥ阻害活性の評価＞
＜１＞本発明のＡＣＥ阻害剤のＡＣＥ阻害作用は、山本
らの方法（日本胸部疾患誌、vol.18, p297-303(1980)）
に準じて以下の方法により評価した。Example 2 <Evaluation of ACE Inhibitory Activity><1> The ACE inhibitory action of the ACE inhibitor of the present invention was determined by the method of Yamamoto et al. (Japanese Chest Disease Journal, vol. 18, p297-303 (1980)).
The evaluation was performed according to the following method.
【００４５】ここで、酵素基質は、Bz(ベンジル)-Gly-H
is-Leu(86mgをリン酸緩衝液16mLに溶解した溶液)を、酵
素溶液は、アンジオテンシンＩ変換酵素（ウサギ肺由
来、Ａ６７７８（シグマ社製））を100mMのホウ酸緩衝
液（pH8.3）に、0.1 units/mLになるように溶解したも
のを使用した。Here, the enzyme substrate is Bz (benzyl) -Gly-H.
is-Leu (a solution of 86 mg dissolved in 16 mL of phosphate buffer), the enzyme solution is angiotensin I converting enzyme (derived from rabbit lung, A6778 (manufactured by Sigma)) 100 mM borate buffer (pH 8.3) The solution used was dissolved in 0.1 units / mL.
【００４６】上記の酵素基質を100μL、酵素溶液を12μ
L及び所定濃度の試料（本発明のＡＣＥ阻害剤）を混合
し、水で全体を250μLとした後、37℃で30分間反応を行
った。100 μL of the above enzyme substrate and 12 μL of the enzyme solution
L and a sample of a predetermined concentration (ACE inhibitor of the present invention) were mixed, and the whole was adjusted to 250 μL with water, and then reacted at 37 ° C. for 30 minutes.
【００４７】反応は1N HCl 250μLを用いて終了させ
た。反応終了液に酢酸エチル1.5mLを入れ、15秒攪拌
し、それを遠心分離した。酢酸エチル層から1.0mLをと
り出して、酢酸エチルを留去し、それに1.0mLの蒸留水
を入れて残渣を溶解し、抽出された馬尿酸の紫外吸収22
8nmの吸光度（OD228）を測定した。阻害率は試料を混合
せず阻害剤なしで反応したときの吸光度を100％とし、
反応時間０分のときの吸光度を０％として求め、阻害率
50％の時の試料の濃度IC50（μM）を決定した。
＜２＞結果
（１）実施例１＜１＞で得られたルビスコの加水分解物
のＡＣＥ阻害作用を評価した。上記実施例２＜１＞にお
いて、試料として実施例１＜１＞で得られたルビスコの
加水分解物を用いた。The reaction was terminated with 250 μL of 1N HCl. 1.5 mL of ethyl acetate was added to the reaction-terminated liquid, and the mixture was stirred for 15 seconds and then centrifuged. Take 1.0 mL from the ethyl acetate layer, distill off the ethyl acetate, add 1.0 mL of distilled water to dissolve the residue, and absorb the extracted hippuric acid by UV absorption.
Absorbance at 8 nm (OD 228 ) was measured. For the inhibition rate, the absorbance when reacting without mixing the sample and the inhibitor is 100%,
The absorbance at 0 minutes of reaction time was calculated as 0%, and the inhibition rate
The concentration IC 50 (μM) of the sample at 50% was determined. <2> Results (1) Example 1 The ACE inhibitory action of the hydrolyzed rubisco obtained in <1> was evaluated. In the above Example 2 <1>, the hydrolyzate of rubisco obtained in Example 1 <1> was used as a sample.
【００４８】その結果、表２で示すように、何れの酵素
を使用したルビスコの加水分解物も、ＡＣＥ阻害作用を
有することが認められた。As a result, as shown in Table 2, it was confirmed that the hydrolyzate of rubisco using any of the enzymes had an ACE inhibitory action.
【００４９】[0049]
【表２】
表２ ルビスコの加水分解物のＡＣＥ阻害作用
──────────────────────────────────
使用酵素 IC50（μg/mL）
──────────────────────────────────
ペプシン ６４
ペプシン−パンクレアチン ７２
──────────────────────────────────
（２）実施例１＜２＞で得られたＡＣＥ阻害ペプチドの
ＡＣＥ阻害作用を評価した。上記実施例２＜１＞におい
て、試料として実施例１＜２＞で得られたＡＣＥ阻害ペ
プチドを用いた。[Table 2] Table 2 ACE inhibitory effect of hydrolyzed rubisco ────────────────────────────────── Enzyme IC 50 (μg / mL) ────────────────────────────────── Pepsin 64 Pepsin-Pancreatin 72 ─ ───────────────────────────────── (2) The ACE-inhibiting peptide obtained in Example 1 <2> The ACE inhibitory effect was evaluated. In the above-mentioned Example 2 <1>, the ACE-inhibiting peptide obtained in Example 1 <2> was used as a sample.
【００５０】その結果、表３で示すIC50値を示した。As a result, the IC 50 values shown in Table 3 were shown.
【００５１】[0051]
【表３】 表３ ＡＣＥ阻害ペプチドのIC50値 ────────────────────────────────── アミノ酸配列 IC50（μg/mL） ────────────────────────────────── ＭＲＷＲＤ ２．２ ＭＲＷ ０．６４ ──────────────────────────────────Table 3 Table 3 IC 50 values of ACE-inhibiting peptides ─────────────────────────────────── Amino acid sequence IC 50 (μg / mL) ────────────────────────────────── MRWRD 2.2 MRW 0.64 ── ────────────────────────────────
【００５２】[0052]
【実施例３】＜自然発症性高血圧ラット（ＳＨＲ）への
単回投与試験＞
＜１＞１９及び２０週齢のＳＨＲに対して、ＡＣＥ阻害
ペプチド（ＭＲＷ）は、２０mg/kg（ＳＨＲの体重）、
ＡＣＥ阻害ペプチド（ＭＲＷＲＤ）は、３０mg/kg（Ｓ
ＨＲの体重）となるように生理食塩水に溶解させて投与
し、その後の血圧の変化を室町機械製MK-2000を用いて
２時間ごとに非観血的に測定した。[Example 3] <Single administration test to spontaneously hypertensive rats (SHR)><1> ACE inhibitory peptide (MRW) was 20 mg / kg (body weight of SHR) against 19- and 20-week-old SHR. ),
ACE inhibitory peptide (MRWRD) was 30 mg / kg (S
It was dissolved in physiological saline and administered so as to be (HR body weight), and the change in blood pressure thereafter was measured noninvasively every 2 hours using MK-2000 manufactured by Muromachi Kikai.
【００５３】＜２＞結果
ＭＲＷ（２０mg/kg）の経口投与により、投与２時間後
の測定において、投与前よりも約１５mmHgの降圧作用が
認められた（図３）。これに対して、生理食塩水を投与
した対照においては、収縮期血圧に変化はなかった。ま
た、４時間後の測定時には、ＭＲＷ投与による降圧作用
は保持されていなかった。<2> Results After oral administration of MRW (20 mg / kg), a hypotensive effect of about 15 mmHg was observed in the measurement 2 hours after administration, as compared with before administration (FIG. 3). In contrast, the systolic blood pressure did not change in the control administered with physiological saline. Further, when measured 4 hours later, the antihypertensive effect of MRW administration was not retained.
【００５４】一方、ＭＲＷＲＤ（３０mg/kg）の投与で
は、投与後２時間後及び４時間後において、投与前より
も約１０mmHgの降圧作用が認められた（図４）。６時間
後においては、収縮期血圧は、投与前のレベルに戻って
いたが、ＭＲＷＲＤは、ＭＲＷと比較して、降圧の程度
は弱いものの、作用の継続時間が長いことが示された。
これは、体内のペプチダーゼによってＭＲＷＲＤが代謝
されて遊離するＭＲＷが、ＡＣＥの真の阻害剤であり、
ＭＲＷＲＤがプロドラッグタイプの阻害剤である可能性
がある。On the other hand, with the administration of MRWRD (30 mg / kg), a hypotensive effect of about 10 mmHg was observed at 2 hours and 4 hours after administration, as compared with before administration (FIG. 4). After 6 hours, the systolic blood pressure had returned to the level before the administration, but MWRRD was shown to have a longer duration of action, although the degree of hypotension was weaker than that of MRW.
This is because MRW, which is metabolized and released from MRWRD by peptidase in the body, is a true inhibitor of ACE,
MRWRD may be a prodrug type inhibitor.
【００５５】[0055]
【発明の効果】本発明により、新規なＡＣＥ阻害剤、及
び該ＡＣＥ阻害剤を有効成分として含有する医薬用組成
物又は食品用組成物を提供することができた。INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a novel ACE inhibitor and a pharmaceutical composition or food composition containing the ACE inhibitor as an active ingredient.
【００５６】[0056]
【配列表】 [Sequence list]
【図面の簡単な説明】[Brief description of drawings]
【図１】実施例１において、ＯＤＳカラムによるＡＣＥ
阻害物質の分画を説明する図。FIG. 1 shows the ACE using an ODS column in Example 1.
The figure explaining the fraction of an inhibitor.
【図２】（ａ）実施例１において、画分IIのＰｈＡカラ
ムによる分画を説明する図。
（ｂ）実施例１において、画分IIのＣＮカラムによる分
画を説明する図。
（ｃ）実施例１において、画分IIのＮ−ＰｈＡカラムに
よる分画を説明する図。FIG. 2 (a) is a view for explaining fractionation of Fraction II by PhA column in Example 1. (B) The figure explaining the fractionation by the CN column of the fraction II in Example 1. (C) The figure explaining the fractionation by the N-PhA column of the fraction II in Example 1.
【図３】実施例３において、ＭＲＷの経口投与がＳＨＲ
の血圧に及ぼす影響を説明する図。FIG. 3 shows that in Example 3, SHR was obtained by oral administration of MRW.
The figure explaining the influence which it has on blood pressure.
【図４】実施例３において、ＭＲＷＲＤの経口投与がＳ
ＨＲの血圧に及ぼす影響を説明する図。[Fig. 4] In Example 3, oral administration of MRWRD was S.
The figure explaining the influence which HR has on blood pressure.
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.7 識別記号 ＦＩ テーマコート゛(参考） Ｃ１２Ｎ 9/99 Ｃ１２Ｐ 21/00 Ｚ Ｃ１２Ｐ 21/00 Ａ６１Ｋ 37/64 Ｆターム(参考） 4B018 MD22 ME04 MF12 4B064 AG21 CA21 CB06 CD20 CE10 DA01 4C084 AA02 AA07 BA01 BA08 BA16 BA23 BA43 DC40 NA14 ZA421 ZC202 4H045 AA10 AA20 AA30 BA12 BA13 CA30 DA57 EA23 FA16 FA70 GA21 GA25 ─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. 7 Identification code FI theme code (reference) C12N 9/99 C12P 21/00 Z C12P 21/00 A61K 37/64 F term (reference) 4B018 MD22 ME04 MF12 4B064 AG21 CA21 CB06 CD20 CE10 DA01 4C084 AA02 AA07 BA01 BA08 BA16 BA23 BA43 DC40 NA14 ZA421 ZC202 4H045 AA10 AA20 AA30 BA12 BA13 CA30 DA57 EA23 FA16 FA70 GA21 GA25
Claims (9)
アンジオテンシンＩ変換酵素を阻害する作用を有するペ
プチド。1. A peptide having the amino acid sequence of SEQ ID NO: 1 and having an action of inhibiting angiotensin I converting enzyme.
３で表される配列を有する、請求項１に記載のペプチ
ド。2. Amino acid Nos. 1 to 1 in SEQ ID NO: 1
The peptide according to claim 1, which has a sequence represented by 3.
求項１に記載のペプチド。3. The peptide according to claim 1, having the amino acid sequence of SEQ ID NO: 2.
を有効成分として含有するアンジオテンシンＩ変換酵素
阻害剤。4. An angiotensin I converting enzyme inhibitor containing the peptide according to any one of claims 1 to 3 as an active ingredient.
キシラーゼ／オキシゲナーゼをペプシンで又はペプシン
とパンクレアチンで加水分解して得られる加水分解物を
有効成分として含有する、請求項４に記載のアンジオテ
ンシンＩ変換酵素阻害剤。5. The angiotensin I conversion according to claim 4, which comprises a hydrolyzate obtained by hydrolyzing ribulose-1,5-bisphosphate carboxylase / oxygenase with pepsin or with pepsin and pancreatin as an active ingredient. Enzyme inhibitors.
の生理的に許容される塩の１種又は２種以上、及び／又
は、請求項５に記載の加水分解物及び／又はその生理的
に許容される塩の１種又は２種以上を有効成分として含
有するアンジオテンシンＩ変換酵素阻害剤を、有効成分
として含有する医薬用組成物。6. One or more kinds of the peptide according to claim 4 and / or a physiologically acceptable salt thereof, and / or the hydrolyzate according to claim 5 and / or a physiological salt thereof. A pharmaceutical composition containing, as an active ingredient, an angiotensin I-converting enzyme inhibitor containing, as an active ingredient, one or two or more of the salts acceptable for the above.
の生理的に許容される塩の１種又は２種以上、及び／又
は、請求項５に記載の加水分解物及び／又はその生理的
に許容される塩の１種又は２種以上を有効成分として含
有するアンジオテンシンＩ変換酵素阻害剤を、有効成分
として含有する食品用組成物。7. One or more kinds of the peptide according to claim 4 and / or a physiologically acceptable salt thereof, and / or the hydrolyzate according to claim 5 and / or a physiological salt thereof. A food composition comprising as an active ingredient an angiotensin I-converting enzyme inhibitor, which comprises one or more salts acceptable for the above as an active ingredient.
キシラーゼ／オキシゲナーゼをペプシンで又はペプシン
とパンクレアチンで加水分解して得られる加水分解物か
らアンジオテンシンＩ変換酵素を阻害する作用を有する
ペプチドを採取することを特徴とするアンジオテンシン
Ｉ変換酵素を阻害する作用を有するペプチドの製造方
法。8. A peptide having an action of inhibiting angiotensin I converting enzyme is collected from a hydrolyzate obtained by hydrolyzing ribulose-1,5-bisphosphate carboxylase / oxygenase with pepsin or with pepsin and pancreatin. A method for producing a peptide having an action of inhibiting angiotensin I converting enzyme, which comprises:
する作用を有するペプチドが請求項１〜３の何れかに記
載のペプチドである、請求項８に記載のペプチドの製造
方法。9. The method for producing a peptide according to claim 8, wherein the peptide having an action of inhibiting the angiotensin I converting enzyme is the peptide according to any one of claims 1 to 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002110456A JP4179586B2 (en) | 2002-04-12 | 2002-04-12 | Novel peptide, angiotensin I converting enzyme inhibitor containing the same, pharmaceutical composition and food composition containing such angiotensin I converting enzyme inhibitor |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002110456A JP4179586B2 (en) | 2002-04-12 | 2002-04-12 | Novel peptide, angiotensin I converting enzyme inhibitor containing the same, pharmaceutical composition and food composition containing such angiotensin I converting enzyme inhibitor |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008057964A3 (en) * | 2006-11-02 | 2008-08-28 | Coca Cola Co | High-potency sweetener composition with rubisco protein, rubiscolin, rubiscolin derivatives, ace inhibitory peptides, and combinations thereof, and compositions sweetened therewith |
| JP2009242335A (en) * | 2008-03-31 | 2009-10-22 | Kagome Co Ltd | Anxiolytic agent and medicine |
| US9101160B2 (en) | 2005-11-23 | 2015-08-11 | The Coca-Cola Company | Condiments with high-potency sweetener |
-
2002
- 2002-04-12 JP JP2002110456A patent/JP4179586B2/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9101160B2 (en) | 2005-11-23 | 2015-08-11 | The Coca-Cola Company | Condiments with high-potency sweetener |
| WO2008057964A3 (en) * | 2006-11-02 | 2008-08-28 | Coca Cola Co | High-potency sweetener composition with rubisco protein, rubiscolin, rubiscolin derivatives, ace inhibitory peptides, and combinations thereof, and compositions sweetened therewith |
| JP2010508822A (en) * | 2006-11-02 | 2010-03-25 | ザ・コカ−コーラ・カンパニー | High intensity sweetener composition having rubisco protein, rubiscoline, rubiscoline derivative, ACE inhibitory peptide, and combinations thereof, and sweetened composition thereby |
| US8017168B2 (en) | 2006-11-02 | 2011-09-13 | The Coca-Cola Company | High-potency sweetener composition with rubisco protein, rubiscolin, rubiscolin derivatives, ace inhibitory peptides, and combinations thereof, and compositions sweetened therewith |
| AU2007317459C1 (en) * | 2006-11-02 | 2013-12-05 | The Coca-Cola Company | High-potency sweetener composition with rubisco protein, rubiscolin, rubiscolin derivatives, ace inhibitory peptides, and combinations thereof, and compositions sweetened therewith |
| JP2009242335A (en) * | 2008-03-31 | 2009-10-22 | Kagome Co Ltd | Anxiolytic agent and medicine |
Also Published As
| Publication number | Publication date |
|---|---|
| JP4179586B2 (en) | 2008-11-12 |
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