CN102432670B - 蚕蛹蛋白源二肽ss及其用途 - Google Patents
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Abstract
本发明公开了一种蚕蛹蛋白源二肽SS，该蚕蛹蛋白源二肽SS的氨基酸序列为Ser-Ser。该蚕蛹蛋白源二肽SS的用途是：作为ACE抑制肽。
Description
技术领域
本发明属于生物技术领域，特别涉及一个能与血管紧张素转换酶结合，抑制其活性的二肽Ser-Ser(SS)。
背景技术
血管紧张素转换酶(Angiotensin converting enzyme，ACE，EC3.4.15.1，文献中曾用名有kininaseIl，dipeptidyl carboxypeptidase I等)是一种二羧肽酶，是导致高血压的一个关键酶，它通过水解作用把血管紧张肽I转化成血管紧张肽II，与此同时，ACE还可钝化血管舒缓激肽，这两种作用均可导致血管收缩，从而引起高血压。因此ACE被认为是引起高血压的一个重要因素。 经研究发现血管紧张素转换酶抑制剂(ACEI)，可通过抑制ACE的活性而达到降血压的作用。ACE抑制剂广泛用于治疗心血管、高血压、心力衰竭、肾衰竭等疾病。ACE抑制剂最初是从蛇毒中发现的，随后人们发现从食物原料中提取的ACE抑制肽，如明胶、酪蛋白、鱼、无花果树胶、α-玉米蛋白等都可作为制备ACE抑制肽的原料。
发明内容
本发明要解决的技术问题是提供一种蚕蛹蛋白源二肽及其用途。
为了解决上述技术问题，本发明提供一种蚕蛹蛋白源二肽SS，该蚕蛹蛋白源二肽SS的氨基酸序列为Ser Ser。
本发明还同时提供了上述蚕蛹蛋白源二肽SS的用途：作为ACE抑制肽。
本发明的蚕蛹蛋白源二肽，其来源于蚕蛹蛋白Bmb000010序列，此二肽是上述序列中的59-60、96-97、251-252、267-268、682-683、886-887、1047-1048、1154-1155、1253-1254、1306-1307等位点的两个氨基酸残基组成。
本发明的二肽的获得途径是：
1、基于药效团模型的ACE抑制肽的筛选：
药效团是一种与生物活性相关的抽象分子特征的表示方法。这些抽象分子特征包括三维的(亲疏水性基团、带电/可离子化基团、氢键供体/受体等)，二维的(子结构)和一维的(物化和生物性质)。本发明利用23个已经报道的二肽结构和ACE抑制活性具有一定差别的化合物数据集构建三维药效团模型(表1)。
表1、训练集肽库
| 序号 | 二肽序列 | IC50(μmol/L) | pIC50 |
| 1 | VW | 1.4 | 5.6635 |
| 2 | IY | 2.1 | 5.2581 |
| 3 | YW | 10 | 3.6974 |
| 4 | DG | 12 | 3.5151 |
| 5 | VY | 10 | 3.6974 |
| 6 | RY | 10.5 | 3.6486 |
| 7 | AF | 15.2 | 3.2787 |
| 8 | LY | 18 | 3.1096 |
| 9 | FY | 25 | 2.7811 |
| 10 | VK | 13 | 3.4351 |
| 11 | MF | 45 | 2.1933 |
| 12 | LW | 50 | 2.0880 |
| 13 | YN | 51 | 2.0682 |
| 14 | SF | 130 | 1.1325 |
| 15 | GY | 259 | 0.4432 |
| 16 | FP | 315 | 0.2474 |
| 17 | LF | 349 | 0.1449 |
| 18 | IR | 696 | -0.5453 |
| 19 | YP | 720 | -0.5793 |
| 20 | QK | 885 | -0.7856 |
| 21 | GK | 2500 | -1.8240 |
| 22 | GS | 3800 | -2.2428 |
| 23 | SG | 8500 | -3.0478 |
本发明所采用的数据处理软件为MOE(Molecular Operating Environment)，通过MOE软件的AUTOGPA模块对训练集进行定量构效分析，所得训练集的分子叠合结果如图1。从叠合结果获得训练集的药效团模型为：2个氢键受体，1个芳香环中心和1个氢键给体(如图2)。通过上述所得药效团模型，进行验证数据集的活性预测，验证药效团模型的预测能力，结果通过10个已经报道的二肽结构的ACE活性验证，结果证明模型的r2＝0.8378，说明药效团模型具有理想的预测能力，用pIC50值代表活性值的大小，pIC50＝6-logIC50(如表2和图3)。
表2 验证集肽库
| 序号 | 二肽序列 | IC50(μmol/L) | pIC50 | $pIC50 |
| 1 | VW | 1.4 | 5.6635 | 5.7873 |
| 2 | AW | 15.4 | 3.2656 | 4.9016 |
| 3 | AA | 51.4 | 2.0604 | 0.7336 |
| 4 | GP | 66 | 1.8103 | -0.9967 |
| 5 | AP | 270 | 0.4016 | 0.8752 |
| 6 | FP | 315 | 0.2474 | 0.5412 |
| 7 | AG | 2500 | -1.8240 | -0.5164 |
| 8 | GK | 5400 | -2.5942 | -2.9616 |
| 9 | AH | 9000 | -3.1050 | -4.9316 |
| 10 | GD | 9200 | -3.1270 | -3.3091 |
注：$pIC50代表的是活性的预测值，与pIC50实测值的大小作比较。
2、虚拟肽库中特定二肽的活性预测：
将Bmb000010蛋白的序列输入PeptideCutter(http://web.expasy.org/peptide_cutter/)模拟酶解软件，利用软件提供的所有蛋白酶进行复合酶解，从酶解所得一系列不同长度肽段中选择出其中的二肽结构，SS是其中之一。
通过药效团模型对二肽SS(本发明所述)进行活性预测，所得预测活性为$pIC50＝3.935，对应的IC50＝7.88μmol/L，预示这个二肽结构可能具有良好的ACE抑制活性。
3、ACE抑制活性的检测方法。
ACE在37℃，pH值为8.3的条件下催化分解血管紧张素I的模拟物Hippuryl-L-Histidyl-L-Leucine(HHL)产生马尿酸(HA)，该物质在紫外225nm处具有特征吸收峰；当加入ACE抑制剂时ACE对HHL的催化分解作用受到抑制，马尿酸的生成量减少，通过HPLC方法，测定加入抑制剂前后所生成马尿酸的量的变化即可算出抑制活性的大小。
反应体系为：依次分别加入20μL 0.1U/mL的ACE、50μL ACE抑制肽(即SS二肽)在37℃温浴5min，然后加入10μL 5mM的HHL底物启动ACE的催化反应，在37℃振荡水浴30min后加入250μL 1.0moL/L的HCl终止反应，体系溶液过0.45μm滤膜后进行RP-HPLC检测分析马尿酸(HA)的含量。上述同样条件，以50μL 0.1moL/L的硼酸缓冲液中(含0.3moL/L的NaCl，pH＝8.3)代替ACE抑制剂作为空白反应体系。
注：上述ACE、HHL底物均是以0.1moL/L的硼酸缓冲液(含0.3moL/L的NaCl，pH＝8.3)为溶剂。
ACE抑制肽(SS二肽)，以不同浓度溶解在0.1moL/L的硼酸缓冲液中(含0.3moL/L的NaCl，pH＝8.3)中而得。
RP-HPLC检测：溶剂I为0.05％(V/V)三氟乙酸(TFA)和0.05％(v/v)的三乙胺(TTA)溶于去离子水中，溶剂II为100％的色谱纯乙睛。溶剂I与溶剂II的比例为70％∶30％(体积比)，流速为0.5mL/min，检测波长为225nm，检测柱温为30℃。
ACE抑制活性根据下式计算：
I％＝(A-B)/A×100％
A：不加入短肽抑制剂时的马尿酸的峰面积；
B：加入短肽抑制剂时的马尿酸的峰面积；
ACE：1U单位定义为，在标准检测条件下，在37℃，1min时间内催化底物(Hippuryl-L-Histidyl-L-Leucine，HHL)，产生1μM马尿酸所消耗ACE的量。即，为ACE的活性单位。
本发明的优点和积极效果：
1)本发明的二肽具有明确的靶标分子。
利用基于药效团的虚拟筛选方法，获得潜在具有活性的ACE抑制肽，能加速新的活性肽的发明速度，为高血压治疗提供更多活性更强的药物或药物先导物。
2)利用计算机辅助虚拟筛选的活性肽，通过化学合成可以验证其生物活性。
该二肽分子可以抑制血管紧张素转换酶的活性。
依据本发明所述的SEQ ID NO：1所述的氨基酸序列，可委托吉尔生化(上海)有限公司合成，从而获得本发明所述的ACE抑制肽(或简称为二肽SS)。
本发明的ACE抑制肽(或简称为二肽SS)的用法和用量如下：
本发明的二肽为口服型，用量为口服一次0.8g(成人)，每日2～3次。
附图说明
下面结合附图对本发明的具体实施方式作进一步详细说明。
图1是训练集中23个二肽结构的分子叠合结果图。
图2是基于训练集产生的药效团模型图；包括1个氢键受体，1个芳香环中心，1个氢键给体。
图3是预测活性与实际活性的相关性分析图；GPA预测值与6-logIC50实际值的相关系数r＝0.9153，r2＝0.8378
图4是二肽SS在1.0mg/mL浓度的ACE抑制活性色谱图；
图4中：
A为空白，不添加二肽SS的ACE抑制活性色谱图；
B为添加1.0mg/mL浓度的二肽SS的ACE抑制活性色谱图。
图5是二肽SS在70.3mg/mL浓度的ACE抑制活性色谱图；
图5中：
A为空白，不添加二肽SS的ACE抑制活性色谱图；
B为添加70.3mg/mL浓度的二肽SS的ACE抑制活性色谱图。
具体实施方式
实施例1、二肽SS在1.0mg/mL的浓度下的ACE抑制活性：
色谱条件：溶剂I为0.05％三氟乙酸(TFA)和0.05％的三乙胺(TTA)溶于去离子水中(即每升溶剂I中含有0.5mL的三氟乙酸和0.5mL的三乙胺)，溶剂II为100％的色谱纯乙睛。溶剂I与溶剂II的比例为70％∶30％(体积比)，ultimate3000戴安液相色谱仪，色谱柱为waters Symmetry C18 5μm 4.6×250mm，流速为0.5mL/min，进样量10μL，检测波长为225nm，检测柱温为30℃。
活性检测(检测方法同上)，结果色谱图(图4)。此时SS浓度为1.0mg/mL。
结果：
A：不加入短肽抑制剂时的马尿酸的峰面积为10.422mAU，
B：加入短肽抑制剂时的马尿酸的峰面积为2.652mAU。
所以，二肽SS在1.0mg/mL时的ACE抑制活性为74.55％。
实施例2、二肽SS在70.3mg/mL的浓度下的ACE抑制活性：
色谱条件：溶剂I为0.05％三氟乙酸(TFA)和0.05％的三乙胺(TTA)溶于去离子水中；溶剂II为100％的色谱纯乙睛。溶剂I与溶剂II的比例为70％∶30％，ultimate3000戴安液相色谱仪，色谱柱为waters Symmetry C18 5μm 4.6×250mm，流速为0.5mL/min，进样量10μL，检测波长为225nm，检测柱温为30℃。
检测方法：将通过化学合成法获得此二肽结构，进行活性检测(检测方法同上)，结果色谱图(图5)。此时SS浓度为70.3mg/mL。
结果：
A：不加入短肽抑制剂时的马尿酸的峰面积为10.422mAU，
B：加入短肽抑制剂时的马尿酸的峰面积为0.650mAU。
所以，二肽SS在70.3mg/mL时的ACE抑制活性为93.76％。
通过实施例1和实施例2中的抑制浓度和活性数据，通过药物抑制浓度计算中的LOGIT法，得出SS的IC50值为0.104mg/mL，既541μmol/L。所得实际验证活性值是理想的，此二肽结构未见报道，属于新的ACE抑制活性肽。
本发明的二肽SS，活性比IR、YP、QK、GK、GS、SG等这些活性二肽的ACE抑制活性都强。
最后，还需要注意的是，以上列举的仅是本发明的若干个具体实施例。显然，本发明不限于以上实施例，还可以有许多变形。本领域的普通技术人员能从本发明公开的内容直接导出或联想到的所有变形及依此序列进行的结构衍生，均应认为是本发明的保护范围。
Claims (1)
1.蚕蛹蛋白源二肽SS的用途，该蚕蛹蛋白源二肽SS 的氨基酸序列为Ser-Ser；其特征是：在制备血管紧张素转换酶抑制剂中的应用。
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Non-Patent Citations (6)
| Title |
|---|
| Angiotensin-1-converting enzyme inhibitory peptides:Chemical feature based pharmacophore generation;Wang Z;《European Journal of Medicinal Chemistry》;20110512;第46卷(第8期);3428-3433 * |
| Wang Z.Angiotensin-1-converting enzyme inhibitory peptides:Chemical feature based pharmacophore generation.《European Journal of Medicinal Chemistry》.2011,第46卷(第8期),3428-3433. |
| 一种源于蚕蛹蛋白组分的新活性肽分离鉴定及其与ACE作用模式研究;王伟;《中国食品科学技术学会第七届年会论文摘要集》;20101231;30-31 * |
| 周宁.液相色谱-质谱联用技术分析三氯氧磷辅助下丝氨酸和组氨酸的成肽产物.《厦门大学学报(自然科学版)》.2004,第43卷(第4期),摘要. |
| 液相色谱-质谱联用技术分析三氯氧磷辅助下丝氨酸和组氨酸的成肽产物;周宁;《厦门大学学报(自然科学版)》;20040731;第43卷(第4期);摘要 * |
| 王伟.一种源于蚕蛹蛋白组分的新活性肽分离鉴定及其与ACE作用模式研究.《中国食品科学技术学会第七届年会论文摘要集》.2010,30-31. |
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