KR101481416B1 - Angiotensin ⅰconverting enzyme inhibitory peptides - Google Patents
Angiotensin ⅰconverting enzyme inhibitory peptides Download PDFInfo
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- KR101481416B1 KR101481416B1 KR20120115443A KR20120115443A KR101481416B1 KR 101481416 B1 KR101481416 B1 KR 101481416B1 KR 20120115443 A KR20120115443 A KR 20120115443A KR 20120115443 A KR20120115443 A KR 20120115443A KR 101481416 B1 KR101481416 B1 KR 101481416B1
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- South Korea
- Prior art keywords
- pro
- peptide
- ala
- ace
- angiotensin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
본 발명은 안지오텐신 Ⅰ 전환효소 (angiotensin Ⅰ converting enzyme, ACE)의 저해활성을 갖는 천연물 유래 생리활성 펩타이드에 관한 것이다.
상기와 같은 본 발명에 따르면, 오리피부 (duck skin)으로부터 분리 및 정제한 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드 및 이의 용도를 제공함으로써, 천연물에서 유래하여 부작용이 없는 고혈압 예방 또는 치료용 약학조성물 뿐만 아니라 고혈압 예방 또는 개선용 건강기능식품을 제공할 수 있는 효과가 있다.The present invention relates to a physiologically active peptide derived from a natural product having an inhibitory activity of an angiotensin converting enzyme (ACE).
According to the present invention, by providing the peptide having the amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro separated and purified from duck skin and its use, There can be provided a pharmaceutical composition for preventing or treating hypertension as well as a health functional food for preventing or improving hypertension.
Description
본 발명은 안지오텐신 Ⅰ 전환효소 (angiotensin Ⅰ converting enzyme, ACE)의 저해활성을 갖는 천연물 유래 생리활성 펩타이드에 관한 것으로서, 더욱 상세하게는 오리피부 (duck skin)으로부터 분리 및 정제한 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드 및 이의 고혈압 예방 또는 치료를 위한 용도에 관한 것이다.The present invention relates to a physiologically active peptide derived from a natural product having an inhibitory activity of angiotensin I converting enzyme (ACE), and more particularly to a physiologically active peptide derived from Trp-Tyr-Pro- Ala-Ala-Pro and its use for the prevention or treatment of hypertension.
고혈압은 심혈관계 질병, 뇌졸중 및 말기신장병의 전개에 있어 주요 위험인자 중 하나이다. 본태성 (本態性) 고혈압이 전체 고혈압의 약 90%를 차지하며 혈액 내의 안지오텐신 (Angiotensin) I 이 안지오텐신 I 전환효소 (Angiotensin I converting enzyme; ACE)에 의하여 안지오텐신 II로 바뀌게 되고 이 안지오텐신 Ⅱ가 혈압상승을 일으켜 본태성 고혈압이 발생하는 것으로 알려져 있다.Hypertension is one of the major risk factors for the development of cardiovascular disease, stroke and end-stage renal disease. Angiotensin I is converted to angiotensin II by an angiotensin I converting enzyme (ACE) in the blood, and angiotensin II is elevated in blood pressure Which is known to cause essential hypertension.
안지오텐신 I 전환효소 (Angiotensin I converting enzyme)는 효소적 활성화를 위하여 아연과 염화물을 필요로 하는 아연 단백질분해효소 계열에 속한다. 상기 효소는 10개의 펩타이드 (decapeptide, angiotensin Ⅰ)로 구성된 비활성형태의 안지오텐신 I을 8개의 펩타이드 (octapeptide, angiotensin Ⅱ)로 된 잠재적 혈관수축물질인 안지오텐신 II로 전환시킴으로써 그리고 혈관을 확장시켜 혈압을 강하시키는 작용을 하는 브래디키닌의 촉매작용을 비활성화시킴으로써 혈압을 조절하는 생리학적 중요한 역할을 한다 (skeggs, kahn, & shumway, 1957). 따라서 ACE를 저해시키는 것이 고혈압을 조절하는 중요한 치료적 접근이라 사료된다.Angiotensin I converting enzyme belongs to the zinc protease family, which requires zinc and chloride for enzymatic activation. The enzyme is produced by converting an inactive form of angiotensin I composed of 10 peptides (decapeptide, angiotensin I) into angiotensin II, a potential vasoconstrictor of octapeptide (angiotensin II), and by expanding blood vessels to lower blood pressure (Skeggs, kahn, & shumway, 1957), which regulates blood pressure by inactivating the catalytic action of acting Bradykinin. Therefore, inhibition of ACE is considered to be an important therapeutic approach to control hypertension.
한편, 최근 연구는 단백질 가수분해물에 포함된 기능성 물질의 설계 및 생산에 초점이 맞추어져 있으며 (Jeon, Byun, & Kim, 2000; Kim, Byun, Jeon, 1999), 뱀독 (snake venom)이 ACE 저해제로서 효과가 있다는 것이 발견된 이후, 현재 인간의 심장마비 및 고혈압의 치료에 사용되는 ACE 저해제를 합성하기 위한 많은 연구가 시도되었다 (예를 들어, captopril, enalapril, alacepril, lisinopril).Recent studies have focused on the design and production of functional materials in protein hydrolysates (Jeon, Byun, & Kim, 2000; Kim, Byun, Jeon, 1999). Snake venom is an ACE inhibitor Many studies have been attempted to synthesize ACE inhibitors currently used in the treatment of human heart failure and hypertension (for example, captopril, enalapril, alacepril, lisinopril).
그러나, 상기 합성된 약물 중 몇몇이 항고혈압 약품으로 매우 효과적임에도 불구하고 종종 부정적인 부작용을 일으키고 있으며, 그러한 잠재적인 부작용 위험 때문에 엄격한 규제아래 사용되고 있는 실정이다.However, although some of the above synthesized drugs are highly effective as antihypertensive agents, they often cause negative side effects and are being used under strict regulations due to such potential side effects risks.
따라서 기존의 합성 항고혈압 조성물을 대체할 수 있는 천연물에 존재하는 항고혈압 화합물을 규명하기 위한 연구가 활발하게 이루어지고 있으며, 최근 천연물에서 추출한 항고혈압 화합물로 cheese whey, casein, zein, corn gluten, bovine blood plasma 등이 알려졌으나, 이에 대한 항고혈압 효과는 아직 평가된 바 없다.Therefore, researches for identifying antihypertensive compounds present in natural products that can replace conventional synthetic antihypertensive compounds have been actively conducted. Recently, antihypertensive compounds extracted from natural products have been used for the production of cheese whey, casein, zein, corn gluten, bovine blood plasma, etc. have been known, but the antihypertensive effect against it has not been evaluated yet.
한편, 오리고기는 혈중 콜레스테롤 수치를 줄이고 항산화 활성을 나타내기 때문에 건강식품으로 널리 알려져 있다. 더불어, 오리고기를 이용한 식품은 심혈관의 기능을 향상시키는 기능식품으로도 사용되고 있다. On the other hand, duck meat is widely known as a health food because it reduces blood cholesterol levels and exhibits antioxidant activity. In addition, duck meat foods are also used as functional foods to improve cardiovascular function.
또한, 안지오텐신 전환 효소 저해 활성을 갖는 조성물과 관련된 종래기술로는 한국공개특허 특2002-0036981호 (산국 꽃잎의 열수 추출물로부터 저분량 안지오텐신 전환 효소 저해제의 제조방법), 한국등록특허 제10-0447534호 (씀바귀뿌리 열수 추출물로부터 저분자량 안지오텐신 전환 효소 저해제를 제조하는 방법 및 상기 방법에 의해 제조된 저분자량 안지오텐신 전환 효소 저해제를 함유하는 식품) 등이 있으며, 최근 많은 ACE 저해제가 천연물 또는 식품 단백질로부터 분리 및 정제되고, 그것들의 항고혈압 효과는 동물모델 (SHR)을 통하여 평가되었다. 명태 껍질 (Alaska pollack skin)로부터 분리 및 정제된 ACE 저해제 펩타이드에 관하여는 이미 보고된바 있다. 또한, 담륜충 (rotifer)으로부터 분리 및 정제된 ACE 저해제 펩타이드에 관하여는 이미 보고된바 있으며, 그 펩타이드의 아미노산 서열은 Asp-Asp-Thr-Gly-His-Asp-Phe-Glu-Asp-Thr-Gly-Glu-Ala-Met이고, 분자량은 1538 Da이다.In addition, prior art related to compositions having an angiotensin converting enzyme inhibitory activity is Korean Patent Publication No. 2002-0036981 (a method for producing a low-dose angiotensin converting enzyme inhibitor from hot water extract of a flower of a country of origin), Korean Patent No. 10-0447534 (A method of producing a low molecular weight angiotensin converting enzyme inhibitor from the extract of roasted roots, and a food containing the low molecular weight angiotensin converting enzyme inhibitor prepared by the above method). Recently, many ACE inhibitors have been separated from natural products or food proteins And their antihypertensive effects were assessed through animal models (SHR). ACE inhibitor peptides isolated and purified from Alaska pollack skin have already been reported. Gly-Asp-Phe-Glu-Asp-Thr-Asp-Asp-Phe-Gly-Asp-Thr- Gly-Glu-Ala-Met, and the molecular weight is 1538 Da.
이에 본 발명자는 오리피부 (duck skin)으로부터 분리 및 정제한 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드가 안지오텐신 Ⅰ 전환효소 (angiotensin Ⅰ converting enzyme, ACE)의 저해활성을 갖는 것을 실험적으로 확인함으로서 본 발명을 완성하게 되었다.Therefore, the present inventors have found that a peptide having an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro separated and purified from duck skin has an inhibitory activity of angiotensin converting enzyme (ACE) The present invention has been completed.
결국 본 발명의 목적은, 천연물에서 유래하여 부작용이 없는 항고혈압 활성을 가지는 오리피부 (duck skin)으로부터 분리 및 정제한 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드 및 이의 용도를 제공함에 있다.Accordingly, an object of the present invention is to provide a peptide having an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro isolated from duck skin having antihypertensive activity derived from a natural product and having no side effects and its use .
또한, 본 발명의 다른 목적은 안지오텐신 Ⅰ 전환효소 (ACE) 저해 효과를 갖는 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열의 펩타이드를 포함하는 고혈압 예방 또는 치료용 약학조성물을 제공함에 있다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating hypertension comprising a peptide having an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro having an angiotensin converting enzyme (ACE) inhibitory effect.
상기 목적을 달성하기 위하여, 본 발명은 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드을 제공한다.In order to achieve the above object, the present invention provides a peptide having an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro.
상기 펩타이드는 혈압강하 효과, 안지오텐신 Ⅰ 전환효소 (ACE) 저해 효과를 가지는 것을 특징으로 한다.The peptide is characterized by having a blood pressure lowering effect and an angiotensin I converting enzyme (ACE) inhibitory effect.
상기 펩타이드는 오리피부 (duck skin)으로부터 분리한 것을 특징으로 한다.
The peptide is characterized in that it is separated from a duck skin.
또한, 본 발명을 상기 펩타이드를 함유하는 식품조성물을 제공한다.
In addition, the present invention provides a food composition containing the peptide.
또한, 본 발명은 상기 펩타이드를 유효성분으로 함유하는 고혈압 예방 또는 개선용 건강기능식품을 제공한다.
The present invention also provides a health functional food for preventing or ameliorating hypertension containing the peptide as an active ingredient.
또한, 본 발명은 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드를 함유하는 안지오텐신 Ⅰ 전환효소 (ACE) 저해제를 제공한다.
The present invention also provides an angiotensin converting enzyme (ACE) inhibitor comprising a peptide having an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro.
또한, 본 발명은 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드를 함유하는 혈압 강하제를 제공한다.
Further, the present invention provides a hypotensive agent containing a peptide having an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro.
또한, 본 발명은 안지오텐신 Ⅰ 전환효소 (ACE) 저해 효과를 갖는 Trp-Tyr-Pro-Ala-Ala-Pro 아미노산 서열의 펩타이드를 포함하는 고혈압 예방 또는 치료용 약학조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating hypertension comprising a peptide of Trp-Tyr-Pro-Ala-Ala-Pro amino acid sequence having an angiotensin I converting enzyme (ACE) inhibitory effect.
상기 약학조성물에 포함되는 펩타이드는 오리피부 (duck skin)으로부터 분리한 것을 특징으로 한다.The peptide contained in the pharmaceutical composition is characterized in that it is separated from a duck skin.
상기와 같은 본 발명에 따르면, 천연물에서 유래하여 부작용이 없는 항고혈압 활성을 가지는 오리피부 (duck skin)으로부터 분리 및 정제한 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드를 식품 및 약학조성물의 용도로 활용할 수 있는 효과가 있다.According to the present invention as described above, a peptide having an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro separated and purified from duck skin having antihypertensive activity derived from a natural product and having no side effects, And a pharmaceutical composition.
또한, 오리피부 (duck skin)으로부터 분리 및 정제한 안지오텐신 Ⅰ 전환 효소 저해활성이 있는 Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 갖는 펩타이드를 유효성분으로 포함하는 고혈압 예방 또는 치료용 약학조성물 뿐만 아니라 고혈압 예방 또는 개선용 건강기능식품을 제공함으로써, 천연물에서 유래하여 부작용이 없는 약학조성물을 제공할 수 있는 효과가 있다.In addition, the pharmaceutical composition for prevention or treatment of hypertension comprising, as an active ingredient, a peptide having an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro having an angiotensin I converting enzyme inhibitory activity separated and purified from duck skin It is possible to provide a pharmaceutical composition derived from a natural product and free from side effects by providing a health functional food for prevention or improvement of hypertension as well as a composition.
도 1 은 α-chymotrypsin을 효소로 사용한 오리피부 (duck skin) 가수분해물을 고속 단백질 액체 크로마토그래피 (fast protein liquid chromatography, FPLC)한 결과 얻은 분획물 및 그것들의 IC50값을 나타낸 그래프.
도 2a 는 도 1에서의 분획물 E를 역상-고성능 크로마토그래피 (reverse-phase high performance liquid chromatography, RP-HPLC)하여 얻은 분획물 및 그것들의 IC50값을 나타낸 그래프.
도 2b 는 도 2a에서의 분획물 E-Ⅰ을 역상-고성능 크로마토그래피 (reverse-phase high performance liquid chromatography, RP-HPLC)하여 얻은 분획물 및 그것들의 IC50값을 나타낸 그래프.
도 3a 는 도 2b에서의 E-Ⅰ-ⅱ을 역상-고성능 크로마토그래피 (reverse-phase high performance liquid chromatography, RP-HPLC)하여 얻은 분획물 및 그것들의 IC50값을 나타낸 그래프.
도 3b 는 도 3a에서의 E-Ⅰ-ⅱ-Ⅲ을 역상-고성능 크로마토그래피 (reverse-phase high performance liquid chromatography, RP-HPLC)하여 얻은 최종적인 펩타이드 및 그것의 IC50값을 나타낸 그래프.
도 4 는 본 발명에 의한 ACE 저해제의 라인위버-버크 플롯 (Lineweaver??Burk plots)을 도시한 그래프.
도 5a 는 자연발생고혈압쥐 (spontaneously hypertensive rats, SHR)에 ACE 저해제를 주산한 경우 및 captopril을 주사한 경우와 정상 쥐의 수축기 혈압의 변화를 나타낸 그래프.
도 5b 는 자연발생고혈압쥐 (spontaneously hypertensive rats, SHR)에 ACE 저해제를 주산한 경우 및 captopril을 주사한 경우와 정상 쥐의 심장박동수 변화를 나타낸 그래프. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the fractions obtained by fast protein liquid chromatography (FPLC) of duck skin hydrolysates using α-chymotrypsin as an enzyme and their IC 50 values.
FIG. 2A is a graph showing the fractions obtained by reverse-phase high performance liquid chromatography (RP-HPLC) of fraction E in FIG. 1 and IC 50 values thereof. FIG.
Graph showing the fractions and their IC 50 values obtained by high-performance chromatography (reverse-phase high performance liquid chromatography , RP-HPLC) - Figure 2b is a fraction E-Ⅰ in Figure 2a reverse phase.
FIG. 3A is a graph showing the IC 50 values of fractions obtained by reverse-phase high performance liquid chromatography (RP-HPLC) of E-I-II in FIG. 2B.
Graph showing the final peptide and its IC 50 value is obtained by high-performance chromatography (reverse-phase high performance liquid chromatography , RP-HPLC) - Figure 3b E-Ⅰ-ⅱ-Ⅲ the reverse phase in Figure 3a.
Figure 4 is a graph showing Lineweaver ' Burk plots of ACE inhibitors according to the present invention.
FIG. 5A is a graph showing changes in the systolic blood pressure of spontaneously hypertensive rats (SHR), when ACE inhibitors are injected and when captopril is injected, and in normal mice.
FIG. 5B is a graph showing changes in heart rate of spontaneously hypertensive rats (SHR), ACE inhibitors, captopril, and normal rats.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.
한편, 이하에서 오리피부 (duck skin)은 오리의 피부 또는 껍질을 의미하며, 오리를 가공하는 과정에서 발생하는 부산물의 하나인 오리 피부 또는 껍질을 포함한다.
Hereinafter, duck skin refers to the skin or skin of a duck, and includes a duck skin or a skin, which is one of the byproducts generated in the processing of the duck.
실시예 1. 실험재료Example 1. Experimental material
오리피부 (duck skin)은 지역 오리농장 (음성, 한국)에서 구입하였다. 토끼 폐에서 얻은 ACE (angiotensin Ⅰ converting enzyme) 및 ACE의 기질 펩타이드 (히푸릴-히스티딜-류신, hippuryl-histidyl-leucine(HHL)), 소 췌장에서 얻은 알파-키모트립신 (α-chymotrypsin) 및 아세토니트릴 (acetonitrile)은 Sigma Aldrich (St. Louis, MO)에서 구입하였고, 그 외 모든 시약들은 상업적으로 이용가능한 최상급이었다.The duck skin was purchased from a local duck farm (Voice, Korea). The angiotensin I converting enzyme (ACE) and the substrate peptide of ACE (hippuryl-histidyl-leucine (HHL)) obtained from rabbit lungs, α-chymotrypsin obtained from small pancreas Acetonitrile was purchased from Sigma Aldrich (St. Louis, Mo.) and all other reagents were commercially available top class.
실시예 2. 오리피부 효소 가수분해물 제조Example 2 Preparation of Duck Skin Enzyme Hydrolyzate
오리피부 효소 가수분해물을 제조하기 위해, 오리피부를 그라인더 (grinder)를 사용하여 분말로 분쇄한 다음 사용하였다.To prepare duck skin enzymatic hydrolysates, duck skin was ground into powder using a grinder and then used.
오리피부 분말 1.0 ㎏ 당 100 ㎖의 완충용액 (buffer solution)을 첨가한 후 30분간 배양시킨 다음에 9개의 샘플에 9가지 효소 각각을 20 ㎖ (또는 g) 첨가하였고, 상기 9가지 효소는 알카레이즈 (alcalase), 콜라겐네이즈 (collaganase), 플라보자임 (flavourzyme), 뉴트레이즈 (neutrase), 파파인 (papain), 펩신 (pepsin), 프로타멕스 (protamex), 트립신 (trypsin) 및 알파-키모트립신 (α-chymotrypsin)이다.100 ml of a buffer solution was added per 1.0 kg of duck skin powder, and the mixture was incubated for 30 minutes. Then, 20 ml (or g) of each of 9 enzymes was added to 9 samples, a protein selected from the group consisting of alcalase, collaganase, flavorzyme, neutrase, papain, pepsin, protamex, trypsin and alpha-chymotrypsin α-chymotrypsin).
효소 가수분해 반응은 최적의 가수분해 수준을 달성하기 위해 8시간 동안 수행하였고, 그 후 즉시 효소를 비활성화하기 위해 100 ℃에서 10 분 동안 가열하였다. 그 다음 얼음을 이용하여 신속하게 20 내지 25 ℃로 냉각시켰으며, 최종적인 효소 가수분해물은 3일 동안 감압하에 동결건조하여 얻었다.The enzymatic hydrolysis reaction was carried out for 8 hours to achieve optimal hydrolysis levels and then immediately heated to 100 < 0 > C for 10 minutes to inactivate the enzyme. It was then rapidly cooled to 20-25 ° C using ice and the final enzymatic hydrolyzate was obtained by lyophilization under reduced pressure for 3 days.
실험예 1. ACE 저해 활성 측정Experimental Example 1. Measurement of ACE inhibitory activity
ACE 저해 활성을 쿠시만 및 청 (Chshman DW, & Cheung, 1971)의 방법을 활용하여 측정하였다.ACE inhibition activity was measured using the method of Kushman and Chung (Chshman DW, & Cheung, 1971).
샘플용액 50 ㎕를 ACE 용액 (25 munits/㎖) 50 ㎕와 섞은 후 37 ℃에서 5 분간 전배양하였고, 상기 혼합물을 150 ㎕의 기질 (pH 8.3에서 0.5 M의 염화나트륨 [NaCl]을 포함한 50 mM의 나트륨 붕산염 [sodium borate buffer] 하의 8.3 mM Hip-His-Leu)과 혼합한 후 동일 온도에서 60분간 더 배양하였다. 마지막으로, 1.0 M HCl 250 ㎕를 첨가하여 반응을 정지시켰고, 1.5 ㎖의 에틸아세테이트 (ethylacetate)로 추출하여 결과적으로 히프루산 (hippuric acid)을 얻었다. 그 다음 800 g으로 15 분간 원심분리한 후 상층액 1 ㎖는 테스트튜브에 옮긴 후 2 시간 동안 실온에서 진공ㅇ증발시켰다. 상기 히프루산을 3.0 ㎖의 증류수에 녹인 후, UV-분광광도계 (UV-spectrophotometer, Varian Inc., Australia)를 사용하여 228 nm에서 흡광도를 측정하였다. IC50값은 ACE 활성을 50% 억제하는데 필요한 억제제의 농도로 정의하였다.50 μl of the sample solution was mixed with 50 μl of ACE solution (25 munits / ml), followed by preincubation at 37 ° C for 5 minutes. The mixture was added to 150 μl of a substrate (pH 8.3 at 50 mM containing 0.5 M sodium chloride [NaCl] 8.3 mM Hip-His-Leu under sodium borate buffer) and incubated for 60 min at the same temperature. Finally, 250 μl of 1.0 M HCl was added to quench the reaction and extracted with 1.5 ml of ethylacetate, resulting in hippuric acid. After centrifugation at 800 g for 15 minutes, 1 ml of the supernatant was transferred to a test tube and then vacuum-evaporated at room temperature for 2 hours. The hyphrulic acid was dissolved in 3.0 ml of distilled water and the absorbance was measured at 228 nm using a UV-spectrophotometer (Varian Inc., Australia). The IC 50 value was defined as the concentration of inhibitor required to inhibit ACE activity by 50%.
상기 실시예 2.에 의해 제조된 가수분해물의 항고혈압 활성은 상술한 ACE의 억제분석을 통하여 평가되었으며, 9개의 효소를 사용한 가수분해물 중에서 α-chymotrypsin을 효소로 사용한 가수분해물이 가장 높은 ACE 저해 활성을 나타내는 것으로 확인되었고, IC50값은 1.72 ㎎/㎖이었다.The antihypertensive activity of the hydrolyzate prepared in Example 2 was evaluated by the inhibition assay of ACE described above. Among the hydrolysates using 9 enzymes, the hydrolyzate using? -Chymotrypsin as the enzyme showed the highest ACE inhibitory activity , And the IC 50 value was 1.72 mg / ml.
실험예 2. 오리피부로부터 항고혈압 펩타이드의 정제Experimental Example 2: Purification of antihypertensive peptides from duck skin
ACE 저해 활성이 있는 펩타이드를 정제하기 위해서는 오리피부의 α-chymotrypsin 가수분해물이 30-K MWCO, 10-K MWCO 및 5-KMWCO 멤브레인 (membranes)을 통해 순차적으로 필터링되어야 하며, 오리피부 단백질의 TFF system에서 저분자량 분획물 (5-KMWCO 이하)이 고분자량 분획물보다 높은 ACE 억제 활성을 보였고, IC50값은 0.816 ㎎/㎖이었다. 한편, 많은 연구결과를 통하여 긴 펩타이드에 비해 짧은 펩타이드가 항산화활성 및 생리활성 효과가 크다는 것이 알려져 있다. 따라서, 5-KMWCO 멤브레인을 통해 얻어진 분획물로부터 항고혈압활성이 있는 펩타이드를 정제하기 위해 하기의 실험을 진행하였다.In order to purify ACE inhibitory peptides, α-chymotrypsin hydrolysates of duck skin must be sequentially filtered through 30-K MWCO, 10-K MWCO and 5-KMWCO membranes, and the TFF system of duck skin protein (Lower than 5-KMWCO) showed higher ACE inhibitory activity than the high molecular weight fractions, and IC 50 value was 0.816 ㎎ / ㎖. On the other hand, it is known from the results of many studies that short peptides have a greater antioxidant activity and physiological activity than long peptides. Therefore, the following experiment was conducted to purify the peptide having antihypertensive activity from the fraction obtained through the 5-KMWCO membrane.
실험예 3. CExperimental Example 3. C 18 18 과 GPC 칼럼 (column)을 사용한 액체 크로마토그래피 (liquid chromatography)And liquid chromatography using a GPC column.
(1) 상기 실험예 2에 의한 오리피부의 α-chymotrypsin 가수분해물 (IC50값= 1.72 ㎎/㎖)을 5-KMWCO 멤브레인으로 필터링하여 얻어진 분획물을 고속 단백질 액체 크로마토그래피 (fast protein liquid chromatography, FPLC)로 분획하였다. 사용조건은 GPC 칼럼 (HiPrep™ 26/60 Sephacryl™ S-100 HR)을 사용하였고, 증류수를 분당 4.0 ㎖의 속도로 흘려주면서 240 nm에서 흡광도를 측정하였다.(1) The fraction obtained by filtering the? -Chymotrypsin hydrolyzate of the duck skin according to Experimental Example 2 (IC 50 value = 1.72 mg / ml) with a 5-KMWCO membrane was subjected to fast protein liquid chromatography (FPLC ). The conditions of use were GPC column (HiPrep ™ 26/60 Sephacryl ™ S-100 HR), and the absorbance was measured at 240 nm while distilled water was flowed at a rate of 4.0 ml per minute.
그 결과 도 1.에 도시한 바와 같이 8개 (A, B, C, D, E, F, G, H)의 분획물이 용출 (elution)되었고, 이를 모두 수집 및 감압하에 동결건조하였다.As a result, fractions of eight (A, B, C, D, E, F, G, H) fractions were eluted as shown in Fig. 1 and all were collected and lyophilized under reduced pressure.
동결건조된 상기 8개의 분획물에 대하여 상기 실험예 1.과 동일한 방법으로 ACE 저해 활성을 측정한 결과 도 1.에 도시한 바와 같이 분획물 E (IC50값= 0.591 ㎎/㎖)가 가장 강력한 ACE 저해 활성을 나타내었다.The ACE inhibitory activities of the eight lyophilized fractions were measured in the same manner as in Experimental Example 1. As shown in Figure 1, fraction E (IC 50 value = 0.591 mg / ml) showed the strongest ACE inhibition Activity.
(2) 상기 분획물 E를 역상-고성능 크로마토그래피 (reverse-phase high performance liquid chromatography, RP-HPLC)를 사용하여 다시 분획하였다. 사용조건은 C18칼럼 (250×20 mm)을 사용하였고, 아세토니트릴 (0-80%)을 분당 4.0 ㎖의 속도로 흘려주면서 215 nm에서 흡광도를 측정하였다.(2) The fraction E was further fractionated using reverse-phase high performance liquid chromatography (RP-HPLC). C 18 column (250 x 20 mm) was used, and absorbance was measured at 215 nm while flowing acetonitrile (0-80%) at a rate of 4.0 ml / min.
그 결과 도 2a.에 도시한 바와 같이 E-Ⅰ, E-Ⅱ, E-Ⅲ, E-Ⅳ, E-Ⅴ 5개의 분획물이 용출되었고, 이를 모두 수집 및 감압하에 동결건조 하였다. As a result, 5 fractions of E-I, E-II, E-III, E-IV and E-V were eluted as shown in FIG. 2A. All the fractions were collected and lyophilized under reduced pressure.
동결건조된 상기 5개의 분획물에 대하여 상기 실험예 1.과 동일한 방법으로 ACE 저해 활성을 측정한 결과 도 2a.에 나타낸 바와 같이 분획물 E-Ⅰ (IC50값= 0.282 ㎎/㎖)이 가장 강력한 ACE 저해 활성을 나타내었다.The ACE inhibitory activity of the 5 freeze-dried fractions was measured in the same manner as in Experimental Example 1. As shown in FIG. 2a, the fraction E-I (IC 50 value = 0.282 mg / ml) Lt; / RTI >
(3) 상기 분획물 E-Ⅰ을 RP-HPLC를 사용하여 다시 분획하였다. 사용조건은 C18칼럼 (250×4.6 mm)을 사용하였고, 아세토니트릴 (0-70%)을 분당 1.0 ㎖의 속도로 흘려주면서 215 nm에서 흡광도를 측정하였다.(3) The fraction E-I was further fractionated using RP-HPLC. The C 18 column (250 x 4.6 mm) was used, and the absorbance at 215 nm was measured while flowing acetonitrile (0-70%) at a rate of 1.0 ml per minute.
그 결과 도 2b.에 나타난 바와 같이 E-Ⅰ-ⅰ, E-Ⅰ-ⅱ, E-Ⅰ-ⅲ, E-Ⅰ-ⅳ 4개의 분획물이 용출되었고, 이를 모두 수집 및 감압하에 동결건조하여, 상기 실험예 1.과 동일한 방법으로 ACE 저해 활성을 측정한 결과 도 2b.에 도시한 바와 같이 E-Ⅰ-ⅱ (IC50값= 0.236 ㎎/㎖)가 가장 강력한 ACE 저해 활성을 나타내었다.As a result, four fractions of E-I-I, E-I-II, E-I-III and E-I-IV were eluted as shown in FIG. 2b. All the fractions were collected and lyophilized under reduced pressure, ACE inhibitory activity was measured in the same manner as in Experimental Example 1. As a result, E-Ⅰ-ⅱ (IC 50 value = 0.236 mg / ml) showed the most potent ACE inhibitory activity as shown in FIG. 2b.
(4) 정제된 펩타이드를 얻기 위해 상기 분획물 E-Ⅰ-ⅱ을 GPC SB-802.5 칼럼 (300×8.0 mm)을 사용하여 RP-HPLC를 실시하였다. 그 결과, 도 3a.에 도시한 바와 같이 E-Ⅰ-ⅱ-Ⅰ, E-Ⅰ-ⅱ-Ⅱ, E-Ⅰ-ⅱ-Ⅲ 3개의 분획물을 수집하였고, 상술한 방법과 동일한 방법으로 ACE 저해 활성을 측정한 결과 도 3a.에 나타난 바와 같이 E-Ⅰ-ⅱ-Ⅲ (IC50값= 0.109 ㎎/㎖)이 가장 강력한 ACE 저해 활성을 보였다.(4) The fraction E-I-ii was subjected to RP-HPLC using a GPC SB-802.5 column (300 x 8.0 mm) to obtain a purified peptide. As a result, three fractions of E-I-II-I, E-I-II-II and E-I-II-III were collected as shown in FIG. As a result of measuring the activity, E-I-II-III (IC 50 value = 0.109 mg / ml) showed the most potent ACE inhibitory activity as shown in FIG.
(5) 또한, 상기 E-Ⅰ-ⅱ-Ⅲ 분획물에 대하여 GPC SB-803 칼럼 (300×3.0 mm)을 사용하여 RP-HPLC를 실시한 결과 최종적으로 정제된 펩타이드(IC50값= 0.095 ㎎/㎖)를 수득하게 되었다.(도 3b)
(5) Further, RP-HPLC was performed on the E-Ⅰ-Ⅱ-Ⅲ fraction using a GPC SB-803 column (300 × 3.0 mm) to obtain a final purified peptide (IC 50 value = 0.095 mg / ) (Figure 3b). ≪ RTI ID = 0.0 >
[표 1][Table 1]
실험예 4. 정제된 펩타이드의 아미노산 서열 확인Experimental Example 4. Identification of Amino Acid Sequence of Purified Peptides
(1) 상기 실험예 3.에 의해 최종적으로 정제된 펩타이드의 정확한 분자량과 아미노산 서열은 한국기초과학연구소 (Korea Basic Science Institute)의 GC/LC Triple Quadrupole Mass Spectrometer (Shinhan Scientific)를 사용하여 결정하였다. 또한 정제된 펩타이드는 NCBI non-redundant peptide database로 검색되었다.(1) The exact molecular weight and amino acid sequence of the peptide finally purified by the above Experimental Example 3 were determined using GC / LC Triple Quadrupole Mass Spectrometer (Shinhan Scientific) of Korea Basic Science Institute. The purified peptides were also screened for the NCBI non-redundant peptide database.
(2) 최종적으로 정제된 ACE 저해 펩타이드의 분자량은 693.90 Da 이고, Trp-Tyr-Pro-Ala-Ala-Pro의 아미노산 서열을 가지는 것으로 확인되었다.
(2) The molecular weight of the finally purified ACE inhibitory peptide was 693.90 Da, and it was confirmed to have an amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro.
실시예 3. 펩타이드의 합성 및 정제Example 3. Synthesis and purification of peptides
펩타이드는 ASP48S (Peprton Inc)를 사용하여 Fmoc SPPS (solid phase peptide synthesis)에 의해 합성하였다. 그리고 펩타이드의 정제는 Vydac Everest C18 column (250 mm × 22 mm, 10 μm)을 사용하여 RP-HPLC에 의하였다.The peptides were synthesized by Fmoc SPPS (solid phase peptide synthesis) using ASP48S (Peprton Inc). Purification of the peptides was done by RP-HPLC using a Vydac Everest C 18 column (250 mm × 22 mm, 10 μm).
용출은 0.1% (v/v)의 trifluoroacetic acid를 포함하는 water-acetonitrile linear gradient (3~40% (v/v)의 acetonitrile)로 실시되었다.Elution was carried out with a water-acetonitrile linear gradient (3-40% (v / v) acetonitrile) containing 0.1% (v / v) trifluoroacetic acid.
실험예 5. ACE 저해 패턴 규명Experimental Example 5. Identification of ACE Inhibition Pattern
상기 실험예 3.에 의해 오리피부로부터 분리 및 정제된 ACE 저해 펩타이드의 억제 메카니즘은 라인위버-버크 플롯 (Lineweaver??Burk plots)을 사용하여 확인하였다.The inhibition mechanism of ACE inhibitory peptides isolated and purified from duck skin according to Experimental Example 3 was confirmed using Lineweaver-Burk plots.
그 결과, 도 4.에 도시한 바와 같이 경쟁적인 것으로 밝혀졌다. 이는 ACE 활성 부위에 결합하는 기질과 상기 펩타이드가 경쟁한다는 것을 의미한다. As a result, it was proved to be competitive as shown in Fig. This means that the peptide competes with the substrate binding to the ACE active site.
실험예 6. 실험동물의 혈압과 심장박도수의 측정Experimental Example 6. Measurement of Blood Pressure and Heart Rate in Experimental Animals
(1) 실험준비(1) Preparing the experiment
8주령의 수컷 자연발생고혈압쥐 (spontaneously hypertensive rats, SHR)와 정상혈압의 실험용 쥐를 동양생명 (성남, 한국)에서 구입하였고, 이를 건국대학교 동물 연구센터에서 1주일 동안 12시간의 light-dark cycle, labortory chow (표준 실험실 먹이공급) 및 증류수의 임의섭식을 유지하였다. 그 후 임의로 선택된 SHR은 각각 untreated, anti-hypertensive peptide (항고혈압 합성 펩타이드) 및 captopril-treated 그룹으로 나누어 실험하였다.A 8-week-old male spontaneously hypertensive rats (SHR) and normotensive rats were purchased from Tong Yang Life Insurance Company (Seongnam, Korea) , labortory chow (standard lab feeding) and distilled water. The randomly selected SHRs were then divided into untreated, anti-hypertensive peptides and captopril-treated groups, respectively.
(2) SHR에서의 항고혈압 활성 효과 확인(2) Confirming the effect of antihypertensive activity in SHR
1) 실험방법1) Experimental method
SHR에 상기 실시예 3.에 의해 제조한 항고혈압 합성 페타이드를 1.0 mg/kg (체중)의 양으로 정맥주사하였고, captopril도 10 mg/kg (체중)의 양으로 정맥주사하였다. 한편, 정상형압의 쥐는 1.0 mg/kg (체중)의 양으로 생리식염수를 정맥주사하였다.SHR was intravenously injected with 1.0 mg / kg body weight of the anti-hypertensive synthetic petide prepared in Example 3, and captopril was intravenously injected in an amount of 10 mg / kg (body weight). On the other hand, normal-pressure rats were intravenously injected with physiological saline at a dose of 1.0 mg / kg (body weight).
또한, 심장박동은 beat와 beat의 간격에 따라 측정하였고, 수축기 혈압은 쥐를 40 ℃의 챔버에서 10분간 예열한 뒤에 tail-cuff 방법 (UR-5000, Ueda Co. Ltd., Tokyo, Japan)을 이용하여 측정하였다. The heart rate was measured according to the interval between beat and beat, and the systolic blood pressure was measured by using a tail-cuff method (UR-5000, Ueda Co. Ltd., Tokyo, Japan) after preheating the rats in a chamber at 40 ° C for 10 minutes Respectively.
2) 실험결과2) Experimental results
도 5.에 구체적으로 도시한 바와 같이, 실험기간 동안 정상 쥐 (wistar rat)의 수축기 혈압은 약 120 mmHg (110 ?? 131 mmHg)였다.As shown in FIG. 5, the systolic blood pressure of the wistar rat was about 120 mmHg (110 ?? 131 mmHg) during the experimental period.
SHR에서 ACE 저해제 (합성 펩타이드)를 주입한 다음 0.5 시간이 경과한 후에 수축기 혈압이 160 mmHg 로 강하하였고, 주사 후 6 시간이 경과될때까지 지속적으로 혈압이 강하되었다 (도 5a). 또한, ACE 저해제 (합성 펩타이드)를 주사하는 경우 captopril를 주사하는 경우보다 혈압을 더 강하시킬 수 있는 것으로 나타났고, 이러한 차이는 주사 후 6 시간이 경과한 시점에 수축기 혈압을 19% 더 강하하는 것 (P = .037, 1-way ANOVA)으로서 통계학적으로 의미가 있는 차이였다.After 0.5 hour of injection of the ACE inhibitor (synthetic peptide) in the SHR, the systolic blood pressure dropped to 160 mmHg and the blood pressure was continuously lowered until 6 hours after the injection (FIG. 5A). In addition, the ACE inhibitor (synthetic peptide) injection showed a stronger drop in blood pressure than the captopril injection, with a 19% drop in systolic blood pressure at 6 hours after injection (P = .037, 1-way ANOVA).
한편, ACE 저해제의 항고혈압활성은 심장박동수의 변화 및 정맥주사 후 (1.0 mg/kg of body weight) 0.5 시간, 3 시간 및 6 시간 후의 수축기 혈압의 변화를 측정하여 평가하였다. 실험기간 동안 정상 쥐의 심장 박동수 및 수축기 혈압은 변화가 없었다. 그러나, SHR에서 ACE 저해제 (합성 펩타이드)를 정맥주사 한 다음 0.5 시간이 경과한 후에 심장박동수 및 수축기 혈압이 감소하는 것으로 나타났고, 정맥주사한 후 6 시간이 경과한 후에는 심장박동수 및 수축기 혈압이 큰 폭으로 감소하였으며, 이러한 감소의 폭은 captopril을 주사한 경우보다 크게 나타났다.
The antihypertensive activity of ACE inhibitors was evaluated by measuring changes in heart rate and changes in systolic blood pressure after 0.5 hour, 3 hours and 6 hours after intravenous injection (1.0 mg / kg of body weight). During the experimental period, heart rate and systolic blood pressure of normal rats were not changed. However, heart rate and systolic blood pressure decreased after 0.5 hours of intravenous injection of ACE inhibitor (synthetic peptide) in SHR. After 6 hours of intravenous injection, heart rate and systolic blood pressure decreased And the width of this decrease was larger than that of captopril.
상기 실험데이터는 Windows 용 SPSS패키지를 사용하여 통계적 유의미성에 대해 평가하였다. 실험값은 평균ㅁ표준 오차 (SE)로 표현되었다. 평균값은 p는<0.05의 수준에서 의미있는 치료수단사이의 차이를 측정하기 위해 Duncan's multiple range test에 의한 변량분석 (analysis of variance, ANOVA)을 사용하여 비교하였다.
The experimental data were evaluated for statistical significance using the SPSS package for Windows. The experimental values were expressed as mean ㅁ standard error (SE). Mean values were compared using a Duncan's multiple range test of analysis of variance (ANOVA) to determine the difference between meaningful treatment measures at a level of p <0.05.
덧붙여서, 최근 한 연구에서 돼지의 골격근 단백질 가수분해물로부터 ACE 저해 펩타이드를 분리한 바 있다. 그 연구에서 밝힌 ACE 저해 펩타이드는 Met-Asn-Pro, Thr-Asn-Pro, Asn-Pro-Pro 및 Ile-Thr-Thr-Asn-Pro의 아미노산 서열을 가지고 있다 (Naskashima, Arihara, Sasaki, Ishikawa, & Itoh, 2002). 또한, 종래 ACE 저해 펩타이드가 오리 고기 단백질의 가수분해물로부터 정제된 바 있다. 상기 정제된 펩타이드의 아미노산 서열은 Glu-Asp-Leu-Glu 이다 (Kim, Kim, & Song, 2003). 상기에서 살펴본 최근의 연구결과에서 알 수 있듯이 대부분의 ACE 저해 펩타이드는 적어도 하나의 Proline residues을 가지고 있고, 본 발명에 의해 정제된 펩타이드도 또한 Proline residues을 포함하고 있다. 또한, 일부의 di- 및 tri- 펩타이드는 aromatic amino acid residues (Trp or Typ) 뿐만 아니라 Proline 또는 Histidine을 포함하여 강력한 항산화 활성 효과를 보여준다. 한편, 본 발명에서 정제한 ACE 저해 펩타이드는 N-terminal에 Trp-Tyr를 포함하고 있다.
In addition, a recent study has isolated ACE inhibitory peptides from skeletal muscle protein hydrolysates of pigs. The ACE inhibitory peptides identified in the study have amino acid sequences of Met-Asn-Pro, Thr-Asn-Pro, Asn-Pro-Pro and Ile-Thr-Thr-Asn-Pro (Naskashima, Arihara, Sasaki, Ishikawa, & Itoh, 2002). In addition, conventional ACE inhibitory peptides have been purified from hydrolysates of duck meat proteins. The amino acid sequence of the purified peptide is Glu-Asp-Leu-Glu (Kim, Kim, & Song, 2003). As can be seen from the above-mentioned recent studies, most of the ACE inhibitory peptides have at least one proline residues, and the peptides purified by the present invention also include proline residues. In addition, some di- and tri-peptides show strong antioxidant activity including proline or histidine as well as aromatic amino acid residues (Trp or Typ). Meanwhile, the ACE inhibitory peptide purified in the present invention contains Trp-Tyr at the N-terminal.
상술한 본 발명에 의해 정제된 펩타이드에 대한 실험결과 및 최근 연구결과들을 고려해 볼 때 본 발명에 의해 정제된 펩타이드가 효과적인 ACE 저해제로 활용될 수 있음을 보여준다. 또한, 정제된 펩타이드의 ACE 저해 활성은 실험적으로 확인한 결과 captopril보다 우수한 것으로 나타났다.
Considering the experimental results of the peptides purified according to the present invention and recent research results, it is shown that the peptides purified by the present invention can be used as effective ACE inhibitors. In addition, ACE inhibitory activity of the purified peptides was found to be superior to captopril by experiment.
이상, 본 발명내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 실시태양일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다.
Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
<110> KONKUK UNIVERSITY INDUSTRIAL COOPERATION CORP
<120> ANGIOTENSIN CONVERTING ENZYME INHIBITORY PEPTIDES
<130> P12-E221
<140> 10-2012-0115443
<141> 2012-10-17
<160> 1
<170> KopatentIn 2.0
<210> 1
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> PEPTIDE
<400> 1
Trp Tyr Pro Ala Ala Pro
1 5
<110> KONKUK UNIVERSITY INDUSTRIAL COOPERATION CORP
<120> ANGIOTENSIN CONVERTING ENZYME INHIBITORY PEPTIDES
<130> P12-E221
<140> 10-2012-0115443
<141> 2012-10-17
<160> 1
<170> Kopatentin 2.0
<210> 1
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> PEPTIDE
<400> 1
Trp Tyr Pro
Claims (10)
A peptide consisting of the amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro.
상기 펩타이드는 혈압강하 효과를 가지는 펩타이드.
The method according to claim 1,
The peptide has a blood pressure lowering effect.
상기 펩타이드는 안지오텐신 Ⅰ 전환효소 (ACE) 저해 효과를 가지는 펩타이드.
The method according to claim 1,
The peptide has an angiotensin I converting enzyme (ACE) inhibitory effect.
상기 펩타이드는 오리피부 (duck skin)으로부터 분리한 것을 특징으로 하는 펩타이드.
The method according to claim 1,
Wherein the peptide is isolated from a duck skin.
A food composition comprising a peptide according to any one of claims 1 to 4.
A health functional food for preventing or improving hypertension comprising the peptide according to any one of claims 1 to 4 as an active ingredient.
Angiotensin I converting enzyme (ACE) inhibitor comprising a peptide consisting of the amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro.
A hypotensive agent comprising a peptide consisting of the amino acid sequence of Trp-Tyr-Pro-Ala-Ala-Pro.
A pharmaceutical composition for preventing or treating hypertension comprising a peptide of Trp-Tyr-Pro-Ala-Ala-Pro amino acid sequence having an angiotensin I converting enzyme (ACE) inhibitory effect.
상기 펩타이드는 오리피부 (duck skin)으로부터 분리한 것을 특징으로 하는 고혈압 예방 또는 치료용 약학조성물.
10. The method of claim 9,
Wherein the peptide is isolated from a duck skin.
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| JP2012046450A (en) | 2010-08-27 | 2012-03-08 | Unitika Ltd | Angiotensin-converting enzyme inhibitory peptide, and method for producing the peptide |
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| US20100093640A1 (en) | 2007-03-06 | 2010-04-15 | Alfred Willy Bonte | Ace-inhibitory peptides from whey and methods for providing the same |
| JP2010037261A (en) | 2008-08-05 | 2010-02-18 | Yamada Bee Farm Corp | Hypotensive peptide |
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