A spontaneous gold(I)-azide alkyne cycloaddition reaction yields gold-peptide bioconjugates which overcome cisplatin resistance in a p53-mutant cancer cell line
A spontaneous gold(I)-azide alkyne cycloaddition reaction yields gold-peptide bioconjugates which overcome cisplatin resistance in a p53-mutant cancer cell line
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DOI：
10.1039/c2sc01127a
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发表时间：
2012-01-01
期刊：
影响因子：
8.4
通讯作者：
Metzler-Nolte, Nils
中科院分区：
文献类型：
--
作者：
Koester, S. David;Alborzinia, Hamed;Metzler-Nolte, Nils
固相肽合成（SPPS）是一种用于组装小到中等大小肽的通用技术，其可以帮助将结合的金属络合物递送到某些细胞区室，例如在癌细胞中。本工作显示了一种新的路线，通过非催化的[ 3 + 2]环加成反应的金叠氮化物与炔基肽的金-肽生物缀合物。合成了具有靶向序列的金（I）四肽缀合物，并在含巯基的生物介质存在下显示出延长的稳定性。它们对所选癌细胞的抗增殖效力（2-50 μ M）对应于缀合物的亲脂性。通过原子吸收光谱法（AAS）测定的Au的细胞摄取表明，高初始摄取等于强细胞毒性。呼吸和酸化率反应后立即与Au-肽共轭物治疗，和基本细胞功能的终端故障是在约10分钟内完成。最多12小时，如通过在微流体生物传感器装置（Bionas传感器芯片系统）中在线监测癌细胞代谢所观察到的。这些Au-肽生物缀合物的作用模式通过各种生化和细胞生物学实验阐明。首先，发现硫氧还蛋白还原酶（TrxR）的强烈选择性抑制，细胞氧化还原过程的调节剂。在这种情况下，发现活性氧（ROS）水平升高，并对分离的小鼠肝线粒体的呼吸产生强烈影响。这些最终导致细胞通过凋亡途径死亡，如流式细胞术，线粒体膜电位（MMP）降低和DNA片段化所示。有趣的是，p53突变型MDA-MB 231乳腺癌细胞中的顺铂抗性可以通过本文提供的Au-肽缀合物克服。
Solid-phase peptide synthesis (SPPS) is a versatile technique for the assembly of small to medium size peptides, that can help in the delivery of bound metal complexes to certain cellular compartments, for example in cancer cells. This work shows a new route to gold-peptide bioconjugates via a non-catalyzed [ 3 + 2] cycloaddition reaction of gold azides with alkynyl peptides. Gold(I) tetrapeptide conjugates with a mitochondria-targeting sequence were synthesized and display prolonged stability in the presence of thiol-containing biological media. Their antiproliferative potency against selected cancer cells (2-50 mu M) corresponds to the lipophilicity of the conjugates. The cellular uptake of Au, determined by atomic absorption spectroscopy (AAS), shows that high initial uptake equals strong cytotoxicity. Respiration and acidification rates react immediately upon treatment with the Au-peptide conjugates, and a terminal breakdown of essential cellular functions is complete within ca. 12 h at most, as observed by online monitoring of the cancer cell metabolism in a microfluidic biosensor device (Bionas sensorchip system). The mode of action of these Au-peptide bioconjugates was elucidated by a variety of biochemical and cell biological experiments. First, a strong selective inhibition of the enzyme thioredoxin reductase (TrxR), a regulator of cellular redox processes, was found. In this context, elevated levels of reactive oxygen species (ROS) and strong effects on the respiration of isolated mouse liver mitochondria were found. These finally lead to cell death via apoptotic pathways, as indicated by flow cytometry, low mitochondrial membrane potential (MMP) and DNA fragmentation. Intriguingly, cisplatin-resistance in p53-mutant MDA-MB231 breast cancer cells could be overcome by the Au-peptide conjugates presented herein.