WO2021197063A1 - 一种具有降血压功效的寡肽及其制备方法与应用 - Google Patents
一种具有降血压功效的寡肽及其制备方法与应用 Download PDFInfo
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- WO2021197063A1 WO2021197063A1 PCT/CN2021/081215 CN2021081215W WO2021197063A1 WO 2021197063 A1 WO2021197063 A1 WO 2021197063A1 CN 2021081215 W CN2021081215 W CN 2021081215W WO 2021197063 A1 WO2021197063 A1 WO 2021197063A1
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- oligopeptide
- blood pressure
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Definitions
- the invention belongs to the technical field of resource utilization of rice wine tanks and blood pressure reduction, and specifically relates to an oligopeptide with the effect of lowering blood pressure, and a preparation method and application thereof.
- Hypertension is the main risk factor for cardiovascular disease, and diet and lifestyle are important ways to prevent hypertension. Although there are differences in various mechanisms for lowering blood pressure, the renin-angiotensin system (Rein-Angiotensin System, RAS) plays a key role in controlling blood pressure, and RAS dysfunction can cause hypertension.
- RAS Rein-Angiotensin System
- Synthetic ACE inhibitors such as captopril and enalapril, are the first class of antihypertensive drugs in the renin-angiotensin system. It can prevent the conversion of inactive angiotensin I into active angiotensin II, thereby blocking the effect of the renin-angiotensin system. But they also inhibit the metabolic degradation of vasodilator kinins including bradykinin, leading to specific side effects, such as allergic reactions, elevated blood potassium levels, dysgeusia, and skin rashes. Unlike most vasodilators, angiotensin-converting enzyme inhibitors (Angiotensin-converting enzyme inhibitors (ACEI) lower blood pressure without causing an increase in heart rate.
- Captopril is the first oral effective non-peptide ACEI. Therefore, inhibitors of ACE inhibitors can not be ignored in lowering blood pressure.
- the primary purpose of the present invention is to provide an oligopeptide with the effect of lowering blood pressure.
- the second object of the present invention is to provide a method for preparing the above-mentioned oligopeptide.
- the third object of the present invention is to provide applications of the above-mentioned oligopeptides.
- An oligopeptide, said antihypertensive peptide having an amino acid sequence as shown in SEQ ID NO:1.
- the present invention uses rice distiller’s grains as the raw material, filtered and centrifuged to remove impurities, guided by the angiotensin-converting enzyme inhibitory activity, and obtained an oligopeptide through multiple reverse liquid chromatography separation and purification.
- the amino acid sequence determination found that the Oligopeptides have as SEQ
- ID NO: 1 The amino acid sequence shown in ID NO: 1, with a molecular weight of 720.857 Da, the inhibitory activity of angiotensin-converting enzyme (ACE) was determined, and it was found that the oligopeptide has a significant inhibitory effect on ACE, and has a significant effect on rice wine waste. Reuse and the application of antihypertensive peptides have certain reference value.
- the oligopeptide has an inhibitory effect on angiotensin converting enzyme.
- the determination of ACE inhibitory activity showed that the IC 50 of the isolated and purified oligopeptide monomer was 280.00 ⁇ 5.77 ⁇ g/mL, which had a good inhibitory effect on angiotensin converting enzyme. At the same time, it shows that the oligopeptide has the effect of lowering blood pressure.
- the present invention also provides a method for preparing the above-mentioned oligopeptides, that is, using rice distiller’s grains as raw materials and obtaining them through separation and purification, specifically: after the rice distiller’s grains are filtered and centrifuged to remove impurities, the angiotensin-converting enzyme inhibitory activity is guided by It is obtained by multiple reverse liquid chromatography separation and purification, and the reverse liquid chromatography separation and purification includes primary separation on a C18 preparative column, secondary separation on a C18 hydrophilic preparative column, and reversed-phase high performance liquid chromatography separation and purification.
- the invention also provides the application of the above-mentioned oligopeptides in the preparation of angiotensin converting enzyme inhibitors.
- the invention also provides the application of the above-mentioned oligopeptides in the preparation of antihypertensive drugs.
- the present invention also provides an angiotensin-converting enzyme inhibitor, including the above-mentioned oligopeptides.
- an angiotensin-converting enzyme inhibitor including the above-mentioned oligopeptides.
- it can also include other adjuvants that can be used in the field of blood pressure reduction.
- the preparation can be prepared into granules. , Capsules and tablets.
- the present invention also provides a blood pressure lowering drug, including the above-mentioned oligopeptide.
- a blood pressure lowering drug including the above-mentioned oligopeptide.
- excipients that can be used in the field of blood pressure reduction can also be included.
- the drug can be prepared in the form of granules, capsules and tablets.
- the present invention uses rice distiller’s grains as a raw material to separate and purify to obtain an oligopeptide with the effect of lowering blood pressure.
- Synthetic, safe and non-toxic, the ACE inhibitory activity test found that the oligopeptide has a significant inhibitory effect on ACE, can be used in the field of lowering blood pressure, and is conducive to the resource utilization of rice wine waste.
- FIG 1 shows the sample connection diagram of the C18 preparative column for one-time separation (D1-5);
- Figure 2 shows the inhibitory activity of C18 preparative column (D1-5) on ACE
- FIG 3 shows the sample connection diagram of the secondary separation of the C18 hydrophilic preparative column (S1-6);
- Figure 4 shows the inhibitory activity of secondary separation samples (S1-6) of C18 hydrophilic preparative column on ACE;
- FIG. 5 is the sample drawing of RP-HCLP separation section (H1-6);
- Figure 6 shows the inhibitory activity of RP-HCLP separated samples (H1-6) on ACE
- Figure 7 is a secondary mass spectrum of component H6
- Figure 8 is a PPSQ sequencing diagram of component H6.
- test methods used in the following experimental examples are conventional methods unless otherwise specified; the materials and reagents used, unless otherwise specified, are commercially available reagents and materials.
- Residual base material (rice distiller's grains) from the fermentation of Shunde Hongli Distillery's rice wine.
- HHL decomposes rapidly under the catalysis of ACE to produce hippuric acid (Hip) and dipeptide (His-Leu, HL).
- Hippuric acid has a maximum absorption at 228nm.
- the ACE enzyme activity is inhibited and the amount of hippuric acid produced decreases. Therefore, the amount of hippuric acid produced can be measured by high performance liquid chromatography to evaluate the inhibition rate of ACEI on ACE activity.
- R the inhibition rate of ACEI samples to ACE (%)
- A The peak area of hippuric acid in the control group
- A0 The peak area of hippuric acid in the blank tube.
- concentration of inhibitor IC 50 is defined as half of ACE inhibition activity under certain conditions require. Since there is not a linear relationship between the inhibition rate and the concentration of the preparation, it is necessary to first draw a curve of the relationship between the inhibitor concentration and the inhibition rate, and then find the IC 50 from the curve.
- oligopeptide material into a 100mg/mL solution, then pass through a 0.45 ⁇ m filter membrane, and use Shimadzu PRC-ODS(K) column (30 mm ⁇ 250 mm, 15 ⁇ m, Shimadzu) chromatographic column for separation and preparation.
- the preparation conditions are as follows: mobile phase A is first-grade water (containing 0.1% TFA), mobile phase B is acetonitrile (containing 0.1% TFA), and the flow rate is 10 mL/min.
- the sample volume is 5 mL, monitored at 245 nm and 280 nm, and the elution procedure is shown in Table 3.
- the sample has about 8 peaks at the commonly used detection wavelength of 214nm for oligopeptides. According to the peak time and peak shape similarity, they are divided into D1, D2, D3, D4 and D5. A total of 5 sections were sampled, and the 5 components were collected separately, and then concentrated and lyophilized in vacuo to reduce the degradation of oligopeptides, and then the inhibitory activity of the 5 components on the ACE enzyme was measured. According to the results of the inhibition rate determination in Figure 2, when the sample concentration is 1 mg/mL, the inhibition rate of the D3 component is the highest, reaching 79.43%, which is selected for the next step of separation and analysis.
- the component D3 with the highest ACE inhibitory activity in the previous step was prepared into a 30mg/mL solution, passed through a 0.45 ⁇ m filter membrane, and then separated and prepared by a C18 hydrophilic preparation column.
- the chromatographic column model was ECOSIL C18 steel column (300 mm ⁇ 20 mm, 10 ⁇ m , Germany), the preparation conditions are: mobile phase A is first-grade water (containing 0.1% TFA), mobile phase B is acetonitrile (containing 0.1% TFA), flow rate is 10mL/min, injection volume is 4mL, at 245nm and 280nm Monitoring, the elution procedure is shown in Table 4.
- the component S2 with the highest ACE inhibitory activity in the previous step was prepared into a 20mg/mL solution, passed through a 0.45 ⁇ m filter membrane, and then separated and prepared by RP-HCLP.
- the chromatographic column model was ECOSIL C18 (260 mm ⁇ 4.6 mm, 5 ⁇ m), the analysis conditions are: mobile phase A is first-grade water (containing 0.1% TFA), mobile phase B is chromatography grade acetonitrile (containing 0.1% TFA), the flow rate is 1 mL/min, and the injection volume is 20 ⁇ L, monitored at 245nm and 280nm, the elution program is shown in Table 5.
- the matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MOLDI-TOF-MS/MS) was used to identify the molecular weights of the two purified peptides, and the mass unit of the attached proton was subtracted from the charge-to-mass ratio.
- the relative molecular mass of oligopeptide H6 is 720.857 Da.
- the amino acid sequencer (PPSQ) is used to determine the amino acid composition of the peptide.
- PPSQ can detect the absolute N-terminal sequence of the oligopeptide.
- the sequence of the oligopeptide H6 was finally determined to be Leu-Ile-Ile-Pro-Glu-His (LIIPEH). This is the first time that LIIPEH (hypertensive oligopeptide H6) has been isolated from natural food products, and all oligopeptides have high ACE inhibitory activity.
- the antihypertensive oligopeptide H6 isolated in the present invention has good ACE inhibitory activity, and it has been reversely verified by artificial synthesis that these two oligopeptides do have the effect of lowering blood pressure, and are expected to be applied In the field of lowering blood pressure, it lays the foundation for the future application of natural product antihypertensive peptides. Further, an IC 50 value of oligopeptides isolated after each step will be significantly decreased, IC 50 values of synthetic oligopeptides isolated lower than, that a greater relationship with the purity of oligopeptides.
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Public Health (AREA)
- Heart & Thoracic Surgery (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Cardiology (AREA)
- Pharmacology & Pharmacy (AREA)
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- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
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- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
属于米酒槽资源化利用和降血压技术领域，具体涉及一种具有降血压功效的寡肽及其制备方法与应用，为促进米酒槽的综合利用，从中开发的具有降血压功效的寡肽，以米酒糟为原料，分离纯化得到一种具有降血压功效的寡肽，该寡肽具有如SEQ ID NO：1所示的氨基酸序列，分子量为720.857Da，纯度高，活性好，能人工合成，安全无毒，经ACE抑制活性的测定发现，该寡肽对血管紧张素转化酶具有显著的抑制作用，可以应用于降血压领域，且有利于米酒废料的资源化利用。
Description
本发明属于米酒槽资源化利用和降血压技术领域，具体涉及一种具有降血压功效的寡肽及其制备方法与应用。
高血压是心血管疾病的主要危险因素，饮食和生活方式是预防高血压的重要途径。尽管各种降血压的机制存在差异，但其中肾素-血管紧张素系统（Rein-AngiotensinSystem，RAS）在控制血压方面起着关键性作用，RAS功能失调可引起高血压。
人工合成的ACE抑制剂，例如卡托普利和依那普利，是肾素血管紧张素系统的第一类抗高血压药物。它能阻止无活性血管紧张素I转化为有活性的血管紧张素II，从而阻断肾素-血管紧张素系统的作用。但它们也抑制包括缓激肽在内的血管扩张剂激肽类的代谢降解，导致特定的副作用，例如过敏反应，血钾水平升高，味觉障碍和皮疹等。与大多数血管扩张剂不同，血管紧张素转化酶抑制剂（Angiotensin-converting
enzyme inhibitors，ACEI）降低血压而不引起心率增加。卡托普利（captopril）是第一个口服有效的非肽类ACEI。因此，抑制药ACE抑制剂在降血压方面具有不容忽视的作用。
在我国，稻米资源深度开发与综合利用仍处起步阶段，我国每吨大米糖化后，大概有半吨的残渣没有得到有效利用，研究水平落后于发达国家，致使它们的经济价值未得到充分体现。因此开发大米蛋白和大米肽具有十分广阔的应用前景和空间。
米酒糟作为米酒工业中的主要副产物，鲜米酒糟其蛋白质含量为28.1%（干计），是一种良好的蛋白质资源。目前仅广东顺德酒厂每年产生的米酒糟就有8-9万吨，估计全省每年产米酒糟40万吨以上。
但除少数厂家将其烘干作饲料外，大部分厂家以低价直接出售给邻近的农户作饲料，还有些厂家以废弃物直接排放，既浪费资源又污染环境。作为大米发酵后的产物，米酒糟中含有大量可利用的寡肽及蛋白，因此，米酒槽的综合利用是亟待开发的领域，可大大提高米酒糟的利用价值，减少环境污染。
为了克服上述现有技术的不足，本发明的首要目的是提供一种具有降血压功效的寡肽。
本发明的第二个目的是提供上述寡肽的制备方法。
本发明的第三个目的是提供上述寡肽的应用。
为实现上述目的，本发明所采用的技术方案为：
一种寡肽，所述降血压肽具有如SEQ ID NO：1所示的氨基酸序列。
本发明以米酒糟为原料，经过过滤、离心除杂后，以血管紧张素转化酶抑制活性为导向，通过多次反向液相色谱分离纯化得到一种寡肽，经氨基酸序列测定发现，该寡肽具有如SEQ
ID NO：1所示的氨基酸序列，分子量为720.857Da，经血管紧张素转化酶（Angiotensin-converting enzyme，ACE）抑制活性的测定发现，该寡肽对ACE具有显著的抑制作用，对米酒废料的再利用及降血压肽的应用有一定参考价值。
优选的，所述寡肽对血管紧张素转化酶具有抑制作用。经ACE抑制活性的测定发现，分离纯化得到的寡肽单体的IC
50为280.00±5.77μg/mL，对血管紧张素转化酶具有较好的抑制作用。同时说明所述寡肽具有降血压的功效。
本发明还提供了上述寡肽的制备方法，即以米酒糟为原料，通过分离纯化得到，具体为：将米酒糟经过过滤、离心除杂后，以血管紧张素转化酶抑制活性为导向，通过多次反向液相色谱分离纯化得到，所述反向液相色谱分离纯化包括C18制备柱一次分离、C18亲水制备柱二次分离和反相高效液相色谱分离纯化。
本发明还提供了上述寡肽在制备血管紧张素转化酶抑制剂中的应用。
本发明还提供了上述寡肽在制备降血压药物中的应用。
本发明还提供了一种血管紧张素转化酶抑制剂，包括上述的寡肽，当然，为提高制剂的适用范围，还可以包括其他可应用于降压领域的辅料，该制剂可制备成颗粒剂、胶囊剂和片剂等形式。
本发明还提供了一种降血压药物，包括上述的寡肽。当然，为提高药物的适用范围，还可以包括其他可应用于降压领域的辅料，该药物可制备成颗粒剂、胶囊剂和片剂等形式。
与现有技术相比，本发明的有益效果是：
本发明以米酒糟为原料，分离纯化得到一种具有降血压功效的寡肽，该寡肽具有如SEQ ID NO：1所示的氨基酸序列，分子量为720.857Da，纯度高，活性好，能人工合成，安全无毒，经ACE抑制活性的测定发现，该寡肽对ACE具有显著的抑制作用，可以应用于降血压领域，且有利于米酒废料的资源化利用。
图1为C18制备柱一次分离分段接样图（D1-5）；
图2为C18制备柱一次分离样品（D1-5）对ACE的抑制活性；
图3为C18亲水制备柱二次分离分段接样图（S1-6）；
图4为C18亲水制备柱二次分离样品（S1-6）对ACE的抑制活性；
图5为RP-HCLP分离分段接样图（H1-6）；
图6为RP-HCLP分离样品（H1-6）对ACE的抑制活性；
图7为组分H6的二级质谱图；
图8为组分H6的PPSQ测序图。
下面对本发明的具体实施方式作进一步说明。在此需要说明的是，对于这些实施方式的说明用于帮助理解本发明，但并不构成对本发明的限定。此外，下面所描述的本发明各个实施方式中所涉及的技术特征只要彼此之间未构成冲突就可以相互组合。
下述实验例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等，如无特殊说明，为可从商业途径得到的试剂和材料。
实施例1 降血压肽的分离
（1）材料与试剂
材料：顺德红荔酒厂米酒发酵剩余基料（米酒糟）。
试剂：具体见表1。
（2）ACE抑制活性的测定方法
2.1 反应液的制备
分别向样品管（B）和空白管（A）加入10μLACE溶液（0.25U/mL，ACE酶溶于pH7的磷酸钾缓冲液中），样品管加入10μL ACEI，空白管10μL 缓冲液，37℃温育5min后加入30μL HHL溶液（6.5mmol/mL，HHL溶于pH8.3的0.1mol·L-1硼酸缓冲液，含0.3mol·L-1 NaCl），在37℃条件下反应1h后，后加入80μL 1.0mol/L HCl 中止反应，得到反应液，如表2。
2.2 色谱条件
色谱柱：ECOSIL C18（260
mm×4.6 mm，5μm）。
流动相与色谱条件：乙腈：超纯水=25：75（含0.1%（v/v）TFA），流速：1mL/min；检测波长：228nm；柱温：30℃；进样量：20µL。
2.3 结果计算
原理：HHL在ACE的催化下快速地分解产生马尿酸（Hip）和二肽（His-Leu，HL），马尿酸在228nm处有最大吸收。当加入ACEI样品时，ACE酶的活性受到抑制，马尿酸生成量减少，所以可通过高效液相色谱测定马尿酸的生成量来评估ACEI对ACE活性的抑制率。
计算公式为：
式中：R：ACEI样品对ACE的抑制率（%）；
A：对照组中马尿酸的峰面积；
B：添加ACEI组中马尿酸的峰面积；
A0：空白管中马尿酸的峰面积。
其中，IC
50的定义为在一定条件下抑制ACE酶活性一半时所需要的抑制剂浓度。由于抑制率与制剂浓度之间不是一个线性的关系，因此必须首先绘制抑制剂浓度与抑制率关系的曲线，再从曲线上查出IC
50。
（3）米酒糟原料的预处理
将米酒糟用400目滤布过滤，弃去滤渣，滤液以55℃旋蒸浓缩，然后4000rpm/min离心15min，保留上清液，将上清液以-80℃冷冻干燥72h，得寡肽原料。
（4）C18制备柱一次分离
先将寡肽原料配制成100mg/mL的溶液，然后过0.45μm滤膜，用Shimadzu
PRC-ODS(K)柱（30 mm×250
mm, 15 μm, Shimadzu）色谱柱进行分离制备，制备条件为：流动相A为一级水（含0.1%TFA），流动相B为乙腈（含0.1%TFA），流速为10mL/min，进样量为5mL，于245nm和280nm处监测，洗脱程序如表3所示。
根据图1的分离检测图谱可以看出，在寡肽常用检测波长214nm下，样品大约有8个峰，根据出峰时间及峰形相似度，一共分为D1、D2、D3、D4和D5，共5段接样，分别收集这5个组分，然后即刻进行浓缩及真空冻干，减少寡肽降解，随后测量5个组分对ACE酶的抑制活性。根据图2的抑制率测定结果可知，样品浓度为1mg/mL时，D3组分的抑制率最高，达79.43%，选择其进行下一步的分离和分析。
（5）C18亲水制备柱二次分离
将上一步中对ACE抑制活性最高的组分D3配制成30mg/mL的溶液，过0.45μm滤膜，采用C18亲水制备柱对其进行再分离制备，色谱柱型号为ECOSIL C18钢柱（300
mm×20 mm,10 μm
,Germany），制备条件为：流动相A为一级水（含0.1%TFA），流动相B为乙腈（含0.1%TFA），流速为10mL/min，进样量为4mL，于245nm和280nm处监测，洗脱程序如表4所示。
根据图3的分离检测图谱可以看出，按照出峰情况及分离度，一共分为S1、S2、S3、S4、S5和S6，共6段接样，分别收集这6个组分，浓缩冻干后测量各组分对ACE酶的抑制活性。根据图4的抑制率测定结果可知，经过进一步分离，当样品浓度为1mg/mL时，S2组分的抑制率最高，高达93.68%。为了明确其中起主要作用的高活性肽段，对S2组分进行下一步的分离和分析。
（6）反相高效液相色谱（RP-HCLP）的分离纯化
将上一步中对ACE抑制活性最高的组分S2配制成20mg/mL的溶液，过0.45μm滤膜，利用RP-HCLP进行再分离制备，色谱柱型号为ECOSIL C18（260
mm×4.6 mm，5μm），分析条件为：流动相A为一级水（含0.1%TFA），流动相B为色谱级乙腈（含0.1%TFA），流速为1mL/min，进样量为20μL，于245nm和280nm处监测，洗脱程序为表5。
经过两次分离纯化后的样品纯度依旧有待提升，调整分离程序后，选取其中分离度好，响应值高的色谱峰富集，根据图5所示的HPLC图谱可以看出，根据出峰情况，一共分为H1、H2、H3、H4、H5和H6，共6个峰接样，收集这6个样品，经旋蒸浓缩和冷冻干燥后测定各组分对ACE的抑制率。根据图6的抑制率测定结果可知，组分H5、H6（1mg/mL）的ACE抑制率分别达到89.27%和91.72%，显著高于其它4个组分（
p<0.05）。故对H5、H6两个寡肽进行详尽的后续研究。
（7）降血压寡肽结构的确定
为了研究H6降血压肽的组成，利用基质辅助激光解吸电离飞行时间质谱仪（MOLDI-TOF-MS/MS）鉴定上述两个纯化肽的分子量，由荷质比减去附着的质子的质量单位，得出寡肽H6的相对分子质量为720.857Da。但由于亮氨酸和异亮氨酸分子量相同，仅从质谱图中难以分辨，故再运用氨基酸测序仪（PPSQ）测定肽的氨基酸组成，PPSQ可测出寡肽的N端绝对序列。根据二级质谱图（图7）和氨基酸测序仪分析图（图8），最终确定寡肽H6的序列为Leu-Ile-Ile-Pro-Glu-His（LIIPEH）。这是首次从食物源天然产物中分离出LIIPEH（降血压寡肽H6），且寡肽皆具有高ACE抑制活性。
实验例2 降血压寡肽的人工合成以及ACE的抑制活性评价
为对降血压寡肽H6进行反向验证，并详细表征其在体外对ACE的抑制作用，由上海杰肽生物科技有限公司合成高纯度的降血压寡肽H6（纯度>98%），并以各阶段分离所得的寡肽的活性进行对比，按照ACE抑制活性的测定方法进行ACE的抑制活性评价试验，试验所得IC
50值如表6所示。
根据表6的结果可以看出，本发明分离得到的降血压寡肽H6具有较好的ACE抑制活性，且通过人工合成的手段反向验证这两种寡肽确实具有降血压的功效，有望应用于降血压领域，为今后天然产物降压肽的应用奠定基础。另外，每步分离后寡肽的IC
50值都会显著下降，人工合成寡肽的IC
50值比分离的要低，这与寡肽的纯度有较大关系。
以上对本发明的实施方式作了详细说明，但本发明不限于所描述的实施方式。对于本领域的技术人员而言，在不脱离本发明原理和精神的情况下，对这些实施方式进行多种变化、修改、替换和变型，仍落入本发明的保护范围内。
Claims (10)
- 一种寡肽，其特征在于，所述寡肽具有如SEQ ID NO：1所示的氨基酸序列。
- 根据权利要求1所述的一种寡肽，其特征在于，所述寡肽对血管紧张素转化酶具有抑制作用。
- 根据权利要求1所述的一种寡肽，其特征在于，所述寡肽具有降血压的功效。
- 权利要求1-3任一项所述的一种寡肽的制备方法，其特征在于，以米酒糟为原料，通过分离纯化得到。
- 根据权利要求4所述的制备方法，其特征在于，将米酒糟经过过滤、离心除杂后，以血管紧张素转化酶抑制活性为导向，通过多次反向液相色谱分离纯化得到。
- 根据权利要求5所述的制备方法，其特征在于，所述反向液相色谱分离纯化包括C18制备柱一次分离、C18亲水制备柱二次分离和反相高效液相色谱分离纯化。
- 权利要求1-3任一项所述的一种寡肽在制备血管紧张素转化酶抑制剂中的应用。
- 权利要求1-3任一项所述的一种寡肽在制备降血压药物中的应用。
- 一种血管紧张素转化酶抑制剂，其特征在于，包括权利要求1-3任一项所述的寡肽。
- 一种降血压药物，其特征在于，包括权利要求1-3任一项所述的寡肽。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115144508A (zh) * | 2022-09-02 | 2022-10-04 | 广州市乾相生物科技有限公司 | 一种适用于多种水溶性肽的hplc分离方法 |
| CN115819503A (zh) * | 2022-10-25 | 2023-03-21 | 上海市农业科学院 | 具有降血压功能的大球盖菇咸味活性肽及其应用 |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113321719B (zh) * | 2021-05-20 | 2022-03-18 | 澳优乳业（中国）有限公司 | 一种寡肽及其制备方法与应用 |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004082709A1 (en) * | 2003-03-18 | 2004-09-30 | Suntory Limited | Angiotensin-converting enzyme inhibitory peptides |
| CN101126105A (zh) * | 2007-06-28 | 2008-02-20 | 武汉工业学院 | 一种用大米分离蛋白制备降血压肽的方法 |
| CN101130801A (zh) * | 2007-08-29 | 2008-02-27 | 王丽萍 | 一种降血压活性肽产品的制备 |
| CN109645498A (zh) * | 2018-12-10 | 2019-04-19 | 四川省农业科学院农产品加工研究所 | 具有血管紧张素转化酶抑制活性的多肽提取物及其制备方法和应用 |
| EP3473640A1 (en) * | 2016-06-16 | 2019-04-24 | Sunstar Inc. | Novel tripeptide |
| CN111484545A (zh) * | 2020-03-31 | 2020-08-04 | 华南农业大学 | 一种米酒糟来源的降血压寡肽及其制备方法与应用 |
| CN111548387A (zh) * | 2020-03-31 | 2020-08-18 | 华南农业大学 | 一种具有降血压功效的寡肽及其制备方法与应用 |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2001245547A1 (en) * | 2000-03-13 | 2001-09-24 | Monsanto Technology Llc | Process for producing peptide sequences possessing anti-hypertension activity |
| CN101906133A (zh) * | 2009-06-03 | 2010-12-08 | 北京大学 | 一种玉米血管紧张素转移酶抑制肽的制备方法及其应用 |
| CN110028550B (zh) * | 2016-07-07 | 2022-08-05 | 华东理工大学 | 降血压肽和降血压蛋白质及其应用 |
-
2020
- 2020-03-31 CN CN202010245463.XA patent/CN111548387B/zh active Active
-
2021
- 2021-03-17 WO PCT/CN2021/081215 patent/WO2021197063A1/zh active Application Filing
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004082709A1 (en) * | 2003-03-18 | 2004-09-30 | Suntory Limited | Angiotensin-converting enzyme inhibitory peptides |
| CN101126105A (zh) * | 2007-06-28 | 2008-02-20 | 武汉工业学院 | 一种用大米分离蛋白制备降血压肽的方法 |
| CN101130801A (zh) * | 2007-08-29 | 2008-02-27 | 王丽萍 | 一种降血压活性肽产品的制备 |
| EP3473640A1 (en) * | 2016-06-16 | 2019-04-24 | Sunstar Inc. | Novel tripeptide |
| CN109645498A (zh) * | 2018-12-10 | 2019-04-19 | 四川省农业科学院农产品加工研究所 | 具有血管紧张素转化酶抑制活性的多肽提取物及其制备方法和应用 |
| CN111484545A (zh) * | 2020-03-31 | 2020-08-04 | 华南农业大学 | 一种米酒糟来源的降血压寡肽及其制备方法与应用 |
| CN111548387A (zh) * | 2020-03-31 | 2020-08-18 | 华南农业大学 | 一种具有降血压功效的寡肽及其制备方法与应用 |
Non-Patent Citations (2)
| Title |
|---|
| CHEN JIWANG, LIU SHANSHAN, YE RAN, CAI GUANGXIA, JI BO, WU YONGNING: "Angiotensin-I converting enzyme (ACE) inhibitory tripeptides from rice protein hydrolysate:Purification and characterization", JOURNAL OF FUNCTIONAL FOODS, vol. 5, no. 4, 8 August 2013 (2013-08-08), NL, pages 1684 - 1692, XP055854829, ISSN: 1756-4646, DOI: 10.1016/j.jff.2013.07.013 * |
| YANG XUE , LI YUNLIANG , WANG YUCHENG: "Angiotensin-I-Converting Enzyme Inhibitory Activity of Rice Protein Hydrolysates and its Molecular Weight Distribution", CHINA FEED, no. 3, 5 February 2017 (2017-02-05), pages 23 - 26, XP055854827, ISSN: 1004-3314, DOI: 10.15906/j.cnki.cn11-2975/s.20170307 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115144508A (zh) * | 2022-09-02 | 2022-10-04 | 广州市乾相生物科技有限公司 | 一种适用于多种水溶性肽的hplc分离方法 |
| CN115819503A (zh) * | 2022-10-25 | 2023-03-21 | 上海市农业科学院 | 具有降血压功能的大球盖菇咸味活性肽及其应用 |
| CN115819503B (zh) * | 2022-10-25 | 2024-01-26 | 上海市农业科学院 | 具有降血压功能的大球盖菇咸味活性肽及其应用 |
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