在之前的工作中，我们报道了作为两亲性缀合物的铱（III）（Ir（III））复合物-肽杂化物（IPH-ACs）和作为两亲性缀合物的三蝶烯-肽杂化物（TPH-ACs），并发现这些杂化物含有三个阳离子 KK(K)GG 肽单元通过 C6-C8 烷基连接体诱导细胞凋亡 II，这是 Jurkat 细胞中非凋亡性程序性细胞死亡 (PCD) 类型之一，与之前报道的细胞凋亡不同。该研究的细节表明，IPH-AC（和 TPH-AC）诱导的 Paraptosis II 通过内质网 (ER) 和线粒体的膜融合或束缚进行，Ca2+ 从 ER 转移到线粒体，从而导致Jurkat 细胞中线粒体膜电位 (ΔΨm) 损失​​。然而，Paraptosis II 的详细机制研究仅在 Jurkat 细胞中进行。在目前的工作中，我们决定对 HeLa-S3 和 A549 细胞以及 Jurkat 细胞中的 paraptosis II 进行机制研究，以研究 paraaptosis II 的一般机制。同时，我们设计并合成了用含有环己基丙氨酸的肽功能化的新TPH-AC，据报道，这种肽可以增强肽在线粒体中的定位。我们发现含有环己基丙氨酸的 TPH-AC 可促进 Jurkat、HeLa-S3 和 A549 细胞中的 Paraptosis II 过程。使用线粒体和细胞质中的荧光Ca2+探针、线粒体和ER的荧光染色剂以及Paraptosis II抑制剂的实验结果表明，TPH-ACs几乎同时诱导线粒体中Ca2+的增加以及ER和线粒体之间的膜融合，这表明我们之前关于paraptosis II机制的假设应该被修改。

In previous work, we reported on iridium(III) (Ir(III)) complex-peptide hybrids as amphiphilic conjugates (IPH-ACs) and triptycene-peptide hybrids as amphiphilic conjugates (TPH-ACs) and found that these hybrid compounds containing three cationic KK(K)GG peptide units through C6-C8 alkyl linkers induce paraptosis II, which is one of the nonapoptotic programmed cell death (PCD) types in Jurkat cells and different from previously reported paraptosis. The details of that study revealed that the paraptosis II induced by IPH-ACs (and TPH-ACs) proceeds via a membrane fusion or tethering of the endoplasmic reticulum (ER) and mitochondria, and Ca2+ transfer from the ER to mitochondria, which results in a loss of mitochondrial membrane potential (ΔΨm) in Jurkat cells. However, the detailed mechanistic studies of paraptosis II have been conducted only in Jurkat cells. In the present work, we decided to conduct mechanistic studies of paraptosis II in HeLa-S3 and A549 cells as well as in Jurkat cells to study the general mechanism of paraptosis II. Simultaneously, we designed and synthesized new TPH-ACs functionalized with peptides that contain cyclohexylalanine, which had been reported to enhance the localization of peptides to mitochondria. We found that TPH-ACs containing cyclohexylalanine promote paraptosis II processes in Jurkat, HeLa-S3 and A549 cells. The results of the experiments using fluorescence Ca2+ probes in mitochondria and cytosol, fluorescence staining agents of mitochondria and the ER, and inhibitors of paraptosis II suggest that TPH-ACs induce Ca2+ increase in mitochondria and the membrane fusion between the ER and mitochondria almost simultaneously, suggesting that our previous hypothesis on the mechanism of paraptosis II should be revised.