JPH04159297A - Peptide inhibiting angiotensin i transferase and preparation thereof - Google Patents
Peptide inhibiting angiotensin i transferase and preparation thereofInfo
- Publication number
- JPH04159297A JPH04159297A JP2284301A JP28430190A JPH04159297A JP H04159297 A JPH04159297 A JP H04159297A JP 2284301 A JP2284301 A JP 2284301A JP 28430190 A JP28430190 A JP 28430190A JP H04159297 A JPH04159297 A JP H04159297A
- Authority
- JP
- Japan
- Prior art keywords
- angiotensin
- peptide
- ala
- converting enzyme
- residue
- 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.)
- Pending
Links
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Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、アンジオテンシン■変換酵素阻害ペプチドと
その製造法に関する。より詳しくは、本発明は、血圧を
上昇させる働きのあるアンジオテンシン■をアンジオテ
ンシンＩから変換する酵素であるアンジオテンシン■変
換酵素を阻害するペプチドに関する。DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to an angiotensin-converting enzyme inhibitory peptide and a method for producing the same. More specifically, the present invention relates to a peptide that inhibits angiotensin-converting enzyme, which is an enzyme that converts angiotensin-I from angiotensin-I, which has the effect of increasing blood pressure.
（従来の技術）
高血圧症は、最大血圧が１６０ｍｎＨｇ以上か、最小血
圧が９５閣Ｈｇ以上の状態である。わが国では患者数が
約２０００万人であるといわれ、り患率の多い疾病であ
る。(Prior Art) Hypertension is a condition in which the systolic blood pressure is 160 mnHg or more or the diastolic blood pressure is 95 mnHg or more. It is said that there are approximately 20 million patients in Japan, and it is a disease with a high incidence rate.
高血圧症は、脳出血、脳梗塞、クモ膜下出血、狭心症、
心筋梗塞、腎硬化症、腎不全、網膜静脈閉塞症など広範
囲の臓器にわたってさまざまな合併症を生じることが知
られており、有効な治療薬が望まれている。Hypertension includes cerebral hemorrhage, cerebral infarction, subarachnoid hemorrhage, angina pectoris,
It is known that various complications occur in a wide range of organs, including myocardial infarction, nephrosclerosis, renal failure, and retinal vein occlusion, and effective therapeutic agents are desired.
生体内において血圧を調節するメカニズムの１つとして
、昇圧系であるレニン・アンジオテンシン系と、降圧系
であるカリクレイン・キニン系がある。レニン・アンジ
オテンシン系では酵素レニンが腎臓の前糸球体細胞（Ｊ
−Ｇ細胞）で生成され、血中でレニン基質であるところ
のアンジオテンシンゲンに作用してアンジオテンシン■
を生成する。このアンジオテンシン■をアンジオテンシ
ン■に変換する酵素がアンジオテンシンＩ変換酵素であ
り、生じたアンジオテンシン■は、細動脈に作用して収
縮を起こさせる。One of the mechanisms that regulates blood pressure in vivo is the renin-angiotensin system, which is a pressor system, and the kallikrein-kinin system, which is a blood pressure lowering system. In the renin-angiotensin system, the enzyme renin is released into the preglomerular cells of the kidney (J
- Angiotensin is produced by G cells) and acts on angiotensingen, which is a renin substrate in the blood.
generate. The enzyme that converts this angiotensin (2) to angiotensin (2) is angiotensin I converting enzyme, and the generated angiotensin (2) acts on arterioles to cause them to contract.
また、アンジオテンシン■は副腎皮質にも作用してアル
ドステロンの合成と分泌を促し、腎臓でのＮａの再吸収
を促進し、体液量を保持する働きもある。このようにし
てアンジオテンシン■により血圧が上昇する。Angiotensin ■ also acts on the adrenal cortex to promote the synthesis and secretion of aldosterone, promotes the reabsorption of Na in the kidneys, and has the function of retaining body fluids. In this way, angiotensin ■ increases blood pressure.
一方、カリクレイン・キニン系では、蛋白分解酵素であ
るカリクレインが、基質であるところのキニノーゲンに
作用してキニンを生じる。キニンは血管を拡張させ、血
圧を下げる作用を有するがキニナーゼＨにより分解をう
ける。キニナーゼ■はアンジオテンシン■変換酵素と同
一物質であることが知られている。On the other hand, in the kallikrein-kinin system, the proteolytic enzyme kallikrein acts on the substrate kininogen to produce kinin. Kinin has the effect of dilating blood vessels and lowering blood pressure, but is degraded by kininase H. Kininase ■ is known to be the same substance as angiotensin ■ converting enzyme.
以上のことから、アンジオテンシンＩ変換酵素を阻害す
ることにより、昇圧物質であるアンジオテンシン■の生
成が抑制され、また、降圧物質であるキニンの分解が抑
制され降圧作用を示す。そこで、アンジオテンシンＩ変
換酵素を阻害することによる高血圧の治療が考えられ、
現在までにカプトプリル、エナラプリル、アラセブリル
、デラプリル等が開発されている。また、カゼインやツ
ェイン、ゼラチン、魚肉などに由来するペプチドも、ア
ンジオテンシンＩ変換酵素を阻害する働きがあることが
知られている（特開昭５９−４４３２４号、特開昭６４
−５４９７号）。From the above, by inhibiting angiotensin I converting enzyme, the production of angiotensin (2), which is a pressor substance, is suppressed, and the decomposition of kinin, which is a hypotensive substance, is suppressed, thereby exhibiting a hypotensive effect. Therefore, treatment of hypertension by inhibiting angiotensin I converting enzyme has been considered.
To date, captopril, enalapril, alacebril, delapril, etc. have been developed. Furthermore, peptides derived from casein, zein, gelatin, fish meat, etc. are known to have the effect of inhibiting angiotensin I-converting enzyme (Japanese Patent Laid-Open Nos. 59-44324 and 64).
-5497).
しかし、カブトリルは強力な血圧降下作用を有するが、
合成法で作られ高価であり、安易に入手することは難し
い。その上、用量が不適当であると腎機能障害や低血圧
をもたらす。また、分子内に存在するＳＨ基のため、発
疹や味覚異常を引き起こすとも言われている。カゼイン
やツェイン、ゼラチン、魚肉などに由来するペプチドは
、天然物を原料とするものであり、製造工程で多くの未
利用物を生成し、その処理に真人な費用がかかるだけで
なく、廃棄による環境汚染の心配がある。However, although Cabtril has a strong antihypertensive effect,
It is produced by a synthetic method and is expensive, making it difficult to obtain it easily. Moreover, inappropriate dosage results in renal dysfunction and hypotension. It is also said to cause rashes and taste abnormalities due to the SH group present in the molecule. Peptides derived from casein, zein, gelatin, fish meat, etc. are natural raw materials, and the manufacturing process generates a lot of unused material, which not only costs a lot of money to dispose of, but also wastes money due to disposal. There are concerns about environmental pollution.
（発明が解決しようとする課題）
したがって、安価に入手でき、副作用の少ない天然のア
ンジオテンシンＩ変換酵素阻害物の開発、さらには、そ
の適切な製造法の開発が望まれている。(Problems to be Solved by the Invention) Therefore, it is desired to develop a natural angiotensin I converting enzyme inhibitor that is inexpensively available and has few side effects, and furthermore, to develop an appropriate method for producing the same.
（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種々検討した結
果、本発明を完成するに至った。(Means for Solving the Problems) As a result of various studies to solve the above problems, the present inventors have completed the present invention.
すなわち、本発明は、Ａｌａ−Ｌｅｕ−Ｐｒｏ−Ｈｉｓ
−Ａｌａの構造を有するアンジオテンシンＩ変換酵素阻
害ペプチドであり、その製造法は、アクチンの存在する
動物臓器から溶媒を用いて抽出して得られる抽出液を分
離精製することを特徴とする上記アンジオテンシンＩ変
換酵素阻害ペプチドの製造法、さらに、Ｆｍｏｃ−アラ
ニン、Ｆ＋＋＋ｏｃ−ロイシン、Ｆｍｏｃ −プロリン
、Ｆｍｏｃ−ヒスチジンを固相合成することを特徴とす
る上記アンジオテンシンＩ変換酵素阻害ペプチドの製造
法である。That is, the present invention provides Ala-Leu-Pro-His
An angiotensin I converting enzyme inhibiting peptide having the structure of -Ala, the method for producing the angiotensin I converting enzyme is characterized by separating and purifying the extract obtained by extracting from an animal organ in which actin is present using a solvent. This is a method for producing a converting enzyme-inhibiting peptide, which further comprises solid-phase synthesis of Fmoc-alanine, F+++oc-leucine, Fmoc-proline, and Fmoc-histidine.
本ペプチドは魚介類の内臓から水抽出、加熱した後、抽
出したペプチド含有物を各種クロマトグラフィーの手法
を使って分離することにより得られる。This peptide can be obtained by extracting water from the internal organs of seafood, heating it, and then separating the extracted peptide-containing substances using various chromatography techniques.
本ペプチドの分離は、水抽出液中のペプチドを逆相クロ
マト用充填剤Ｃ＋＠（シリカ担体にオクタデシル基を結
合させたもの）に吸着させ、アセトニトリル１５％溶液
で溶出し、イオン交換用クロマト充填剤ＣＭ（担体にカ
ルボキシメチル基を結合させたもの）に吸着させ、ＰＨ
６，０，７，０゜８．０，９．Ｏの段階的溶出法で得た
両分のうち、アンジオテンシンＩ変換酵素阻害画分をさ
らに、逆相ＨＰＬＣで分離することにより得られる。To separate this peptide, the peptide in the aqueous extract is adsorbed onto reverse phase chromatography packing material C+@ (a silica carrier with an octadecyl group bonded to it), eluted with a 15% acetonitrile solution, and ion exchange chromatography packing. The agent is adsorbed onto CM (carboxymethyl group bonded to a carrier), and the PH
6,0,7,0°8.0,9. Of the two fractions obtained by the O stepwise elution method, the angiotensin I converting enzyme inhibiting fraction is further separated by reverse phase HPLC.
本ペプチドは、動物の体内に広く分布しているアクチン
の一部である。したがって、アクチンの存在する全ての
動物から抽出、分離することが可能である。This peptide is part of actin, which is widely distributed in the animal body. Therefore, it is possible to extract and isolate actin from all animals in which it exists.
また、本ペプチドは、Ｆｍｏｃ（９−フルオレニルメチ
ルオキシカルボニル）−アミノ酸である、Ｆ−ｏｃ−ア
ラニン、Ｆｍｏｃ−ロイシン、Ｆｍｏｃ−プロリン、Ｆ
＋＋＋ｏｃ−ヒスチジンを用いた固相合成法で製造する
こともできる。In addition, this peptide contains Fmoc (9-fluorenylmethyloxycarbonyl)-amino acids, F-oc-alanine, Fmoc-leucine, Fmoc-proline, and Fmoc-proline.
It can also be produced by solid phase synthesis using +++oc-histidine.
（実施例） 以下、実施例により本発明を具体的に説明する。(Example) Hereinafter, the present invention will be specifically explained with reference to Examples.
実施例１
カツオ腸１０６．３ｇに４倍量の水を加え、ホモジナイ
ザーにッセイＡＭ−１型）を用いて、水冷下１０．００
Ｏｒｐｍで５分間ホモジナイズした。Example 1 Add 4 times the amount of water to 106.3 g of bonito intestines, and use a homogenizer (Sssey AM-1 model) to homogenize the intestine for 10.00 g under water cooling.
Homogenized for 5 minutes in Orpm.
１２０°Ｃで５分間オートクレーブした後４０，０００
Ｇで４０分間４℃で遠心分離し、その上清を濃縮後、Ｃ
Ｉ８充填剤（Ｗａｔｅｒｓ社Ｂｕｌｋ ＣＩｓ ５５
〜１０５μｍ）１００ｇを詰めたカラムに通し、ペプチ
ドを吸着させた。このカラムを蒸留水１４３５、ｗｆで
洗浄後、アセトニトリル１５％溶液でペプチドを溶出し
た。この操作により４．２ｇのペプチド画分を得た。40,000 after autoclaving at 120°C for 5 minutes
Centrifuge at 4°C for 40 minutes at G, concentrate the supernatant, and then centrifuge at 4°C.
I8 filler (Waters Bulk CIs 55
~105 μm) was passed through a column packed with 100 g to adsorb the peptide. After washing this column with distilled water 1435, wf, the peptide was eluted with a 15% acetonitrile solution. Through this operation, 4.2 g of peptide fraction was obtained.
本ペプチド画分は、１０ｍＭリン酸バッファー（ｐＨ６
，０）で平衡化したアクセルｐｌｕｓ ＣＭ（Ｗａｔｅ
ｒｓ社）７５ｇを詰めた簡易カラムに添加し、１０ｍＭ
リン酸バッファー（ｐＨ６，Ｏ）１５００ｍ１、 （ｐ
Ｈ７，０）１５００ｍ、 （ｐＨ８゜０）１５００ｄ、
（ｐＨ９，０）１５００戚で溶出した。This peptide fraction was prepared using 10mM phosphate buffer (pH 6).
, 0), the accelerator plus CM (Wate
rs) to a simple column packed with 75 g of 10mM
Phosphate buffer (pH 6, O) 1500ml, (p
H7,0) 1500m, (pH8゜0) 1500d,
It was eluted at pH 9,0 and 1500.
このうちアンジオテンシン■変換酵素阻害能の強い画分
をさらに細分画するため、逆相ＨＰＬＣで分離を行った
。ＨＰＬＣ条件はＨＰＬＣ条件■に示した。Among these, in order to further subdivide the fraction with strong angiotensin-converting enzyme inhibitory ability, separation was performed by reverse phase HPLC. HPLC conditions are shown in HPLC conditions (■).
ＨＰＬＣ条件■
カラム ＲＰ−１８（ｅ） Ｉ Ｘ ２５ Ｃ１ｌ
メルク社検出 ＵＶ ２１０ｎｍ
流速 ４ａｄ！／ｓｉｎ
溶離液 Ａ ： ０．０５％ＴＦＡ
Ｂ ： ０．０５％ＴＦＡ ５０χアセトニトリルＡ液
１００χから、８００分後Ｂ液が
５０χになるような直線グラジェン
ト
条件■では、リテンションタイム１６２分のピークに高
いＡＣＥ阻害活性があった。したがって、このピークを
分取し、条件■で示したＨＰＬＣ条件で、さらに精製し
た。HPLC conditions ■ Column RP-18(e) I X 25 C1l
Merck detection UV 210nm flow rate 4ad! /sin Eluent A: 0.05% TFA B: 0.05% TFA 50χ Acetonitrile Under linear gradient conditions ■ where solution A becomes 100χ and solution B becomes 50χ after 800 minutes, a peak with a retention time of 162 minutes appears. It had high ACE inhibitory activity. Therefore, this peak was fractionated and further purified under the HPLC conditions shown in condition (2).
ＨＰＬＣ条件■
カラム ＲＰ−１８（ｅ） ４ ｍ Ｘ ２５ ｃ
ｍ メルク社検出 ＵＶ ２１０ｎｓ
流速 １．０ｄ／ｓｉｎ
溶離液 Ａ ： ０．０５％ＴＦＡ
Ｂ ： ０．０５％ＴＦＡ ５０χアセトニトリルＡ液
１００χから、８００分後Ｂ液が
５０χになるような直線グラジェン
ト
条件■でのＨＰＬＣにより、本ピークはチャート１ （
第１図）に示したように、１０個のピークに分かれた。HPLC conditions ■ Column RP-18(e) 4 m x 25 c
m Merck detection UV 210ns Flow rate 1.0d/sin Eluent A: 0.05% TFA B: 0.05% TFA 50χ Acetonitrile Linear gradient conditions such that from A solution 100χ to B solution 50χ after 800 minutes By HPLC at ■, this peak was found in Chart 1 (
As shown in Figure 1), it was divided into 10 peaks.
このうち、ピーク４に高いアンジオテンシンｌ変換酵素
阻害活性が認められたので、ピーク４をさらにＨＰＬＣ
条件■で分離した。Among these, peak 4 was found to have high angiotensin l-converting enzyme inhibitory activity, so peak 4 was further analyzed by HPLC.
Separation was performed under condition ■.
ＨＰＬＣ条件■
カラム Ａｓａｈｉｐａｃｋ Ｇ５−２２０ （７，
５ｍｘ５０ｃｍ）旭化成工業株式会社製
溶離液 ５０ｇＭ酢酸アンモニウム
流速 １．０Ｉ１１／ａｅｉｎ
検出 ＵＶ ２１０ｎｗ
チャート１のピーク４をＨＰＬＣ条件■で分離したチャ
ートをチャート２（第２図）に示した。HPLC conditions ■ Column Asahipack G5-220 (7,
5m x 50cm) manufactured by Asahi Kasei Corporation Eluent 50gM ammonium acetate Flow rate 1.0I11/aein Detection UV 210nw Chart 2 (Figure 2) shows a chart in which peak 4 in Chart 1 was separated under HPLC condition (■).
このうち、ピーク３に非常に強いアンジオテンシンｌ変
換酵素阻害活性が認められた。Among these, very strong angiotensin l-converting enzyme inhibitory activity was observed in peak 3.
該ピークをＡｐｐｌｉｅｄ Ｂｉｏｓｙｓｔｅｒ社の気
相プロティン−シーケンサ−（Ｍｏｄｅｌ ４７０Ａ
）とオンラインの高速液体クロマトグラフィーを用いて
Ｅｄｍａｎ分解反応を行い、各サイクルで得られるＰＴ
）（−アミノ酸を同定した。その結果、
Ａｌａ−Ｌｅｕ−Ｐｒｏ−Ｈｉｓ−Ａｌａの構造をもつ
ペンタペプチドであることがわかった。The peak was measured using an Applied Biosyster gas phase protein sequencer (Model 470A).
) and online high-performance liquid chromatography to perform the Edman decomposition reaction, and the PT obtained in each cycle was
) (-amino acid was identified. As a result, it was found to be a pentapeptide with the structure Ala-Leu-Pro-His-Ala.
実施例２
本ペプチドをＦｅ＋ｏｃ（９−フルオレニルメチルオキ
シカルボニル）−アミノ酸（Ｆｍｏｃ−アラニン、Ｆｍ
ｏｃ−ロイシン、Ｆｍｏｃ−プロリン、Ｆｍｏｃ−ヒス
チジン）を用いるペプチド固相合成で合成した。このペ
プチドはＹＭＣ−ＯＤＳ−Ｒ５カラム（株式会社ワイエ
ムシイ製）で０．１％ＴＦＡ （）リフルオロ酢酸）水
溶液のイニシャル液から０．１％ＴＦＡの７０％アセト
ニトリル溶液をファイナル液とする２５分の直線グラジ
ェントで分析（流速１、 ０ａｄ！／ｓｉｎ 、検出Ｕ
Ｖ２１０ｎｍ） シた結果、９８％の純度を示した。Example 2 This peptide was synthesized with Fe+oc(9-fluorenylmethyloxycarbonyl)-amino acid (Fmoc-alanine, Fm
It was synthesized by peptide solid phase synthesis using oc-leucine, Fmoc-proline, Fmoc-histidine). This peptide was analyzed using a YMC-ODS-R5 column (manufactured by YMC Co., Ltd.) in a 25-minute straight line starting from an initial solution of 0.1% TFA (refluoroacetic acid) aqueous solution to a final solution of 70% acetonitrile solution of 0.1% TFA. Analysis with gradient (flow rate 1, 0ad!/sin, detection U
V210nm) The result showed a purity of 98%.
この物質を用いてアンジオテンシンＩ変換酵素を５０％
阻害する濃度であるＩＣｓ。は７９．４ＭＭであった。Using this substance, angiotensin I converting enzyme was converted to 50%
ICs are the inhibiting concentrations. was 79.4MM.
アンジオテンシン、Ｉ変換酵素阻害の測定は、カッシュ
マンらの方法（バイオケミカル・ファーマクロジー２０
巻 １６３７〜１６４８頁（１９７１） ）を改良しり
丸山らの方法（アグリカルチエアル・バイオロジカル・
ケミストリー４６巻５号（１３９３〜１３９４頁（１９
Ｂ２） ）にしたがった。Angiotensin, I-converting enzyme inhibition was measured by the method of Cushman et al. (Biochemical Pharmacology 20
Volume 1637-1648 (1971)) was improved by the method of Shirimaruyama et al.
Chemistry Vol. 46 No. 5 (pp. 1393-1394 (19
B2) ) was followed.
すなわち、試験管に本発明ペプチド含有組成物水溶液３
０μ！と酵素基質としてＬ−ヒプリルヒスチジルロイシ
ン（シグマ社製）とＮａＣＩを含有したｐＨ８，３のホ
ウ酸バッファー２５０μｌを加え、３７℃で１０分間ブ
レインキュベーションした。その後、アンジオテンシン
Ｉ変換酵素含有液１００μｌを加え酵素反応を開始した
。このときホウ酸バッファーの濃度は０．１Ｍ、Ｌ−ヒ
プリルヒスチジルロイシン濃度は５１μｌＭ、ＮａＣ１
３００ｍＭであり、阻害がかからない場合の酵素活性は
８Ｕである。３７℃、ｐＨ８，３で３０分間振動しつつ
反応せしめた後、ｌＮＨＣｌ２５０μｌを加え、反応を
停止させた。酢酸エチル１． ５Ｉ１１を加え、１５秒
間振盪させて酵素反応で生じた馬尿酸を抽出し、２００
０ｒｐｍ、１０分間遠心分離し、酢酸エチル層１．０Ｍ
１を試験管に採取した。酢酸エチルをホットドライパス
の中で１２０℃、３０分間加温し、完全に除去した後、
室温で５分間放置した。そして、ＦＩｚＯ１，Ｏａｄ！
を加え、生成した馬尿酸の量を２２８ｎ−の吸光度を測
定して求めた。酵素反応に使用したアンジオテンシン■
変換酵素含有液は、ラビットラングアセトンパウダー（
シグマ社製）Ｉｇを０．１Ｍホウ酸バッファー（ｐＨ８
，３）１０ｄに溶かし、よく攪拌した後、４°Ｃ１４０
、０００ｇで４０分間遠心分離した上清を０．１Ｍホウ
酸バッファー（ｐＨ８，３）で希釈して作製した。That is, an aqueous solution 3 of the peptide-containing composition of the present invention is placed in a test tube.
0μ! 250 μl of a pH 8.3 boric acid buffer containing L-hypril histidyl leucine (manufactured by Sigma) and NaCI as enzyme substrates were added to the mixture, and the mixture was incubated at 37° C. for 10 minutes. Thereafter, 100 μl of an angiotensin I converting enzyme-containing solution was added to start the enzyme reaction. At this time, the concentration of the boric acid buffer was 0.1M, the concentration of L-hypril histidylleucine was 51μlM, and the concentration of NaC1
At 300 mM, the enzyme activity without inhibition is 8 U. After reacting with shaking at 37° C. and pH 8.3 for 30 minutes, 250 μl of 1NHCl was added to stop the reaction. Ethyl acetate 1. Add 5I11 and shake for 15 seconds to extract hippuric acid produced by the enzyme reaction.
Centrifuge at 0 rpm for 10 minutes, and remove the 1.0M ethyl acetate layer.
1 was collected in a test tube. After heating ethyl acetate in a hot dry path at 120°C for 30 minutes and completely removing it,
It was left at room temperature for 5 minutes. And FIzO1, Oad!
was added, and the amount of hippuric acid produced was determined by measuring the absorbance at 228n-. Angiotensin used in enzyme reaction■
The converting enzyme-containing solution is Rabbit Lang Acetone Powder (
Sigma) Ig was added to 0.1M borate buffer (pH 8).
, 3) Dissolve in 10d, stir well, and then heat to 4°C140
The supernatant obtained by centrifugation at ,000g for 40 minutes was diluted with 0.1M boric acid buffer (pH 8.3).
阻害率は下記の式を使用して求めた。The inhibition rate was determined using the following formula.
（発明の効果）
以上説明したとおり、本発明によれば、従来廃棄されて
いた魚介類内臓から、血圧上昇を押さえる作用を有する
アンジオテンシン１変換酵素阻害ペプチドが得られる。(Effects of the Invention) As explained above, according to the present invention, an angiotensin 1-converting enzyme-inhibiting peptide having the effect of suppressing an increase in blood pressure can be obtained from conventionally discarded fish and shellfish viscera.
第１図は実施例１において、アンジオテンシン２変換酵
素阻害能の強い画分をＨＰＬＣ条件■で分離したチャー
ト、第２図は第１図のピーク４をさらにＨＰＬＣ条件■
で分離したチャートを示す。
（ばか１名）Figure 1 is a chart in which the fraction with strong angiotensin 2-converting enzyme inhibitory ability was separated under HPLC condition ■ in Example 1, and Figure 2 is a chart in which peak 4 in Figure 1 was further separated by HPLC condition ■.
Shows a chart separated by . (1 idiot)
Claims (3)
を有するアンジオテンシン I 変換酵素阻害ペプチド。 （ただし、Ａｌａはアラニン、Ｌｅｕはロイシン、Ｐｒ
ｏはプロリン、Ｈｉｓはヒスチジン残基を示す。(1) Angiotensin I converting enzyme inhibitory peptide having a structure of Ala-Leu-Pro-His-Ala. (However, Ala is alanine, Leu is leucine, Pr
o represents proline and His represents histidine residue.
出して得られる抽出液を分離、精製することを特徴とす
るＡｌａ−Ｌｅｕ−Ｐｒｏ−Ｈｉｓ−Ａｌａの構造をも
つアンジオテンシン I 変換酵素阻害ペプチドを製造す
る方法。(2) Angiotensin I converting enzyme inhibitory peptide having the structure of Ala-Leu-Pro-His-Ala, which is characterized by separating and purifying the extract obtained by extracting with a solvent from animal organs where actin exists. How to manufacture.
ｏｃ−プロリン、Ｆｍｏｃ−ヒスチジンを固相合成する
ことを特徴とするＡｌａ−Ｌｅｕ−Ｐｒｏ−Ｈｉｓ−Ａ
ｌａの構造をもつアンジオテンシン I 変換酵素阻害ペ
プチドを製造する方法。(3) Fmoc-alanine, Fmoc-leucine, Fm
Ala-Leu-Pro-His-A characterized by solid phase synthesis of oc-proline and Fmoc-histidine
A method for producing an angiotensin I converting enzyme inhibitory peptide having a structure of la.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2284301A JPH04159297A (en) | 1990-10-24 | 1990-10-24 | Peptide inhibiting angiotensin i transferase and preparation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2284301A JPH04159297A (en) | 1990-10-24 | 1990-10-24 | Peptide inhibiting angiotensin i transferase and preparation thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH04159297A true JPH04159297A (en) | 1992-06-02 |
Family
ID=17676759
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2284301A Pending JPH04159297A (en) | 1990-10-24 | 1990-10-24 | Peptide inhibiting angiotensin i transferase and preparation thereof |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH04159297A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000264845A (en) * | 1999-02-04 | 2000-09-26 | Nippon Synthetic Chem Ind Co Ltd:The | Hypocholesterolemic agent and its use |
-
1990
- 1990-10-24 JP JP2284301A patent/JPH04159297A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000264845A (en) * | 1999-02-04 | 2000-09-26 | Nippon Synthetic Chem Ind Co Ltd:The | Hypocholesterolemic agent and its use |
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