马来酰亚胺基烯二炔化合物对生物大分子的损伤作用

孙可; 鲁浩天; 张梦思; 胡爱国

doi:10.14135/j.cnki.1006-3080.20220530002

马来酰亚胺基烯二炔化合物对生物大分子的损伤作用

doi: 10.14135/j.cnki.1006-3080.20220530002

华东理工大学材料科学与工程学院, 上海市先进聚合物材料重点实验室, 上海 200237

基金项目: 国家自然科学基金（21871080）

详细信息

作者简介:
孙可：孙　可 (1997-)，男，硕士生，主要从事有机小分子合成研究。E-mail：kesunqust@163.com

通讯作者:
胡爱国，E-mail：hagmhsn@ecust.edu.cn

中图分类号: O621.3
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- 文章访问数: 122
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- 被引次数: 0
出版历程
- 收稿日期: 2022-05-30
- 网络出版日期: 2022-06-28

Cleavage of Biological Macromolecules by Maleimide-Based Enediyne Compounds

Shanghai Key Laboratory of Advanced Polymeric Materials, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, China

摘要

摘要: 利用Sonogashira偶联反应合成了3种具有不同羟基数量的马来酰亚胺基烯二炔，它们可以通过马来酰亚胺促进的重排和环芳香化反应（MARACA）作用机制产生高活性的双自由基中间体。电子顺磁共振实验结果证实了体系中自由基中间体的产生。DNA凝胶电泳实验结果证实了这些烯二炔能够在生理温度下对质粒DNA产生裂解作用，并表现出浓度依赖性。蛋白质凝胶电泳实验结果表明烯二炔对蛋白质的降解能力与其产生自由基的能力一致。对于炔丙位上具有杂原子的高反应活性烯二炔结构，其可在低浓度（5 mmol/L）下破坏蛋白质主链结构的完整性致使蛋白质失活。研究结果为探究马来酰亚胺基烯二炔分子对病毒蛋白质等生物大分子的损伤奠定基础，为开发此类分子潜在的临床应用价值提供新的思路。
- 烯二炔 /
- MARACA反应 /
- 双自由基 /
- DNA /
- 蛋白质
Abstract: Three kinds of maleimide-based enediynes with different numbers of hydroxyl groups at the alkyne termini were synthesized by Sonogashira coupling reaction between diiodomaleimide and different kinds of terminal alkynes. Through the maleimide assisted rearrangement and cycloaromatization (MARACA) mechanism, these enediyne molecules can be converted to enyne-allene structures at physiological temperatures, leading to the formation of highly reactive diradical intermediates through Myer-Saito cyclization reactions. Electron paramagnetic resonance experiments confirmed the generation of free radical intermediates in the system by using a free radical trapping agent N-tert-Butyl-α-phenylnitrone. DNA gel electrophoresis experiments confirmed that these enediynes could generate highly reactive free radicals and further abstract hydrogen atoms from the DNA backbone at physiological temperature, resulted in the fragmentation of plasmid DNA strands and cleavage of supercoiled DNA into cyclic form. The DNA cleavage process is concentration-dependent, and different enediyne structures exhibit different cleaving effects under the same concentration gradient. Protein gel electrophoresis experiments showed that the ability of the enediyne compounds to degrade proteins is in line with their ability for generating free radicals. For the highly reactive enediyne structure with an oxygen atom at the propargyl position, the structural integrity of the protein backbone can be destroyed at a rather low concentration (5 mmol/L) of enediyne compounds, resulted in protein denaturing. Altogether, this work provides preliminary results for future research on maleimide-based enediyne for the damaging of a variety of biological macromolecules such as viral proteins, tumor cell DNA, and provides new ideas for the potential clinical applications of these highly intriguing synthetic enediyne compounds.
- enediyne /
- MARACA /
- diradicals /
- DNA /
- protein

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图 1 烯二炔化合物的合成路线

Figure 1. Synthetic route of the enediyne compounds

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图 2 EDY-B和EDY-C的核磁共振氢谱

Figure 2. ¹H-NMR spectra of EDY-B and EDY-C

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图 3 化合物EDY-A（红色）与对照组（黑色）的EPR谱图

Figure 3. EPR spectra of compound EDY-A (red) and control (black)

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图 4 在37 ℃下，化合物EDY-A（a）和EDY-C（b）与pUC19 DNA作用48 h后的凝胶电泳图（单位：mmol/L）

Figure 4. Gel electrophoresis of compound EDY-A (a) and EDY-C (b) reacting with pUC19 DNA at 37 ℃ for 48 h (Unit: mmol/L)

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图 5 不同种类烯二炔化合物对牛血清白蛋白和溶菌酶的SDS-PAGE凝胶电泳图

Figure 5. SDS-PAGE gel electrophoresis of the different enediyne compounds on BSA and lysozyme

(a) c/(mmol·L⁻¹): Lane 1~8: 0.1, 0.2, 0.5, 1, 2, 5, 10, 20;lane 9: EDY-A control group; Con: Protein control group. (b) c /(mmol·L⁻¹): lane 1~5: 1, 2, 5, 10, 20；Con: Protein control group. (c) c /(mmol·L⁻¹): lane 1~5: 1, 2, 5, 10, 20; Con: Protein control group. (d) c/(mmol·L⁻¹): lane 1~7: 1, 2, 5, 7.5, 10, 20, 40；lane 8: EDY-C control group; Con: Protein control group

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