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中国精品科技期刊2020

多肽类ACE抑制剂的设计合成及生物活性

周佳琪 马春燕 李晓晖

周佳琪,马春燕,李晓晖. 多肽类ACE抑制剂的设计合成及生物活性[J]. 食品工业科技,2022,43(23):26−34. doi:  10.13386/j.issn1002-0306.2022030117
引用本文: 周佳琪,马春燕,李晓晖. 多肽类ACE抑制剂的设计合成及生物活性[J]. 食品工业科技,2022,43(23):26−34. doi:  10.13386/j.issn1002-0306.2022030117
ZHOU Jiaqi, MA Chunyan, LI Xiaohui. Design, Synthesis and Bioactivity of Polypeptide ACE Inhibitors[J]. Science and Technology of Food Industry, 2022, 43(23): 26−34. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022030117
Citation: ZHOU Jiaqi, MA Chunyan, LI Xiaohui. Design, Synthesis and Bioactivity of Polypeptide ACE Inhibitors[J]. Science and Technology of Food Industry, 2022, 43(23): 26−34. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022030117

多肽类ACE抑制剂的设计合成及生物活性

doi: 10.13386/j.issn1002-0306.2022030117
详细信息
    作者简介:

    周佳琪(1997−),女,硕士研究生,研究方向:功能性多肽药物的设计与合成,E-mail:jqzhoudut@163.com

    通讯作者:

    李晓晖(1964−),女,博士,教授,研究方向:多肽药物研究,E-mail:lxhxh@dlut.edu.cn

  • 中图分类号: TS201.1

Design, Synthesis and Bioactivity of Polypeptide ACE Inhibitors

  • 摘要: 本研究以血管紧张素I转化酶(angiotensin converting enzyme, ACE)抑制肽PHP1和PHP2为研究母肽,通过替换氨基酸残基改变目标多肽的疏水性、带电性等因素,利用生物信息学工具评估多肽潜在的生物活性,设计了19个多肽类似物。采用多肽固相合成法合成目的多肽类似物,并进行体外生物活性检测。结果显示多肽类似物均具有较高的ACE抑制活性,其中,PHP1A-6(IC50=3.87 μmol/L)、PHP2A-3(IC50=3.33 μmol/L)、PHP2A-4(IC50=2.86 μmol/L)和PHP2A-7(IC50=4.58 μmol/L)的ACE抑制活性最高,较母肽有显著提高(P<0.05),PHP1A-3、PHP1A-4、PHP1A-7、PHP2A-1和PHP2A-10具有同母肽相当的抑制活性,IC50<10 μmol/L。绝大部分多肽类似物的α-葡萄糖苷酶抑制活性与母肽相比均有明显提高,PHP1A-3(IC50=3.09 μmol/L)、PHP1A-7(IC50=9.51 μmol/L)、PHP2A-6(IC50=5.58 μmol/L)、PHP2A-11(IC50=2.35 μmol/L)和PHP2A-12(IC50=3.98 μmol/L)的活性最高。其中,PHP1A-3和PHP1A-7具备较强的ACE抑制和α-葡萄糖苷酶抑制双重活性。含Cys的多肽类似物在1 mg/mL浓度下,ABTS + · 清除率均在85%以上,具备潜在的抗氧化活性。分子对接研究了ACE抑制肽与ACE的构效关系,表明抑制肽可以与ACE的氨基酸残基产生多个稳定的氢键、疏水相互作用、π-π堆积及盐桥,增加了对ACE的抑制作用。
  • 图  1  ACE与多肽类似物的复合物模型

    Figure  1.  The models of ACE in complex with the peptide analogues

    表  1  PHP1、PHP2及其类似物的序列

    Table  1.   Sequence of PHP1, PHP2 and their analogues

    名称序列名称序列
    PHP1Val-Val-Tyr-Pro-Trp-ThrPHP2Thr-Lys-Thr-Tyr-Phe-Pro-His-Phe
    PHP1A-1Cys-Val-Tyr-Pro-Trp-ThrPHP2A-1Thr-Lys-Leu-Tyr-Phe-Pro-His-Phe
    PHP1A-2Val-Cys-Tyr-Pro-Trp-ThrPHP2A-2Ile-Lys-Leu-Tyr-Phe-Pro-His-Phe
    PHP1A-3Val-Val-Tyr-Pro-Trp-PhePHP2A-3Ala-Lys-Leu-Tyr-Phe-Pro-His-Phe
    PHP1A-4Cys-Val-Tyr-Pro-Trp-PhePHP2A-4Ile-Ala-Leu-Tyr-Phe-Pro-His-Phe
    PHP1A-5Val-Cys-Tyr-Pro-Trp-PhePHP2A-5Ala-Ala-Leu-Tyr-Phe-Pro-His-Phe
    PHP1A-6Gly-Val-Tyr-Pro-Trp-PhePHP2A-6Ala-Ala-Leu-Tyr-Phe-Pro-Leu-Phe
    PHP1A-7Val-Ala-Tyr-Pro-Trp-PhePHP2A-7Ala-Lys-Ala-Tyr-Phe-Pro-Arg-Phe
    PHP2A-8Gly-Lys-Leu-Tyr-Phe-Pro-His-Phe
    PHP2A-9Ile-Gly-Leu-Tyr-Phe-Pro-His-Phe
    PHP2A-10Gly-Gly-Leu-Tyr-Phe-Pro-His-Phe
    PHP2A-11Gly-Gly-Leu-Tyr-Phe-Pro-Leu-Phe
    PHP2A-12Gly-Lys-Gly-Tyr-Phe-Pro-Arg-Phe
    注:PHP1/2A(Analogue of Pig Hemoglobin Peptide 1/2,猪血红蛋白源多肽1/2的类似物)。
    下载: 导出CSV

    表  2  PHP1、PHP2及其类似物的理化性质

    Table  2.   Physicochemical properties of PHP1, PHP2 and their analogues

    名称pIGRAVY净电荷脂肪族氨基酸指数非稳定性指数半衰期(h)生物活性预测分数
    mammalian reticulocytesYeast cellE.coli
    PHP15.490.650096.67−13.52100>20>100.50
    PHP1A-15.520.367048.33−26.081.2>20>100.82
    PHP1A-25.490.367048.33−0.95100>20>100.82
    PHP1A-35.491.233096.6711.53100>20>100.83
    PHP1A-45.520.950048.33−1.301.2>20>100.91
    PHP1A-55.490.950048.3324.10100>20>100.98
    PHP1A-65.520.467048.3311.5330>20>100.98
    PHP1A-75.490.833065.0024.10100>20>100.97
    PHP28.29−0.725+10.0019.867.2>20>100.60
    PHP2A-18.29−0.163+148.759.257.2>20>100.76
    PHP2A-28.600.487+197.50−1.36200.5>100.87
    PHP2A-38.640.150+161.259.254.4>20>100.92
    PHP2A-46.741.2000110.0019.86200.5>100.92
    PHP2A-56.780.863073.7519.864.4>20>100.95
    PHP2A-65.571.7380122.5032.834.4>20>100.97
    PHP2A-79.99−0.263+225.0023.404.4>20>100.95
    PHP2A-88.60−0.125+148.75−1.3630>20>100.94
    PHP2A-96.740.925097.5019.86200.5>100.93
    PHP2A-106.740.312048.7535.2930>20>100.96
    PHP2A-115.521.188097.5048.2530>20>100.98
    PHP2A-129.99−0.812+20.00−8.4430>20>100.96
    注:mammalian reticulocytes:哺乳动物的红细胞;Yeast cell:酵母细胞;E.coli:大肠杆菌。
    下载: 导出CSV

    表  3  合成肽的生物活性

    Table  3.   Bioactivity of synthetic peptides

    名称(MW m/z)RP-HPLCACE IC50
    (μmol/L)
    α-Glucosidase IC50
    (μmol/L)
    ABTS+·清除率
    (%)
    Calcd.[M+H]+Rt(min)Purity(%)
    PHP1763.39764.45.55998.316.34±0.378897.09±1.795.80±0.51
    PHP1A-1767.33768.35.25198.6720.25±5.54**116.03±1.09**95.89±0.43**
    PHP1A-2767.33768.25.22799.7336.66±7.90**88.87±1.74**89.76±1.22**
    PHP1A-3809.41810.47.36099.678.48±0.783.09±0.43**8.18±0.69
    PHP1A-4813.35814.57.39197.978.64±1.92104.46±1.79**94.00±0.28**
    PHP1A-5813.35814.47.32998.9513.34±2.2854.49±2.39**85.53±4.26**
    PHP1A-6767.36768.46.98499.843.87±0.21*331.95±2.60**10.10±0.59**
    PHP1A-7781.38782.46.48399.579.67±0.429.51±0.25**12.00±1.21**
    PHP21039.511040.510.16998.048.64±0.14296.79±2.1427.73±0.37
    PHP2A-11051.551052.26.53398.149.48±1.061889.59±1.30**23.28±1.11**
    PHP2A-21063.591064.28.47495.8040.06±2.84**1872.49±1.12**25.73±0.50*
    PHP2A-31021.541022.46.57298.493.33±0.34**306.32±3.76**25.53±1.30*
    PHP2A-41006.531007.59.81797.432.86±0.46**278.23±1.71**28.54±0.69
    PHP2A-5964.48965.18.37098.3558.11±2.30**21.24±2.32**35.43±1.52**
    PHP2A-6940.51941.512.75298.2333.93±2.87**5.58±0.88**37.62±0.28**
    PHP2A-7998.53999.110.99598.154.58±0.57*7949.34±0.96**18.47±1.47**
    PHP2A-81007.521008.46.27298.4815.18±2.72**957.91±1.71**23.14±0.60**
    PHP2A-9992.51993.49.86698.1125.13±1.86**90.82±1.27**30.44±1.15**
    PHP2A-10936.45937.38.19297.738.22±1.021022.05±1.97**28.60±0.84
    PHP2A-11912.47913.210.33999.2911.08±1.472.35±0.27**29.88±1.32*
    PHP2A-12970.50971.34.13499.8724.94±2.71**3.98±0.25**21.49±1.73**
    注:Rt为多肽在液相分析图中的保留时间;PHP1系列设计肽与PHP1比较,PHP2系列设计肽与PHP2比较,“*”代表差异显著(0.01<P<0.05),“**”代表差异极显著(P<0.01)。
    下载: 导出CSV
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  • 收稿日期:  2022-03-10
  • 网络出版日期:  2022-10-20
  • 刊出日期:  2022-11-23

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