Click to enter: activation of oligo-arginine cell-penetrating peptides by bioorthogonal tetrazine ligations
Click to enter: activation of oligo-arginine cell-penetrating peptides by bioorthogonal tetrazine ligations
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DOI：
10.1039/c8sc04394a
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发表时间：
2019-01-21
期刊：
影响因子：
8.4
通讯作者：
Lowik, Dennis W. P. M.
中科院分区：
文献类型：
--
作者：
Bode, Saskia A.;Timmermans, Suzanne B. P. E.;Lowik, Dennis W. P. M.
细胞穿透肽能够跨细胞膜运输多种货物。虽然有希望，但它们通常不被认为是用于治疗目的，因为它们缺乏可控的活性和细胞选择性。我们已经开发了一种激活策略的基础上分裂八精氨酸细胞穿透肽（CPP），可以通过生物正交连接激活。为此，我们制备了两个非穿透的四精氨酸半，用四嗪或用互补的双环[6.1.0]壬炔（BCN）基团官能化。我们证明，一个积极的八精氨酸可以在原位重建后，混合互补的分裂肽。所产生的活化肽与公认的细胞穿透肽八精氨酸一样有效地被吸收。也可以使用反式-环辛烯（TCO）作为连接配偶体来实现寡聚腺苷的活化，而异辛烯对于原位使用而言似乎动力学上太慢。我们进一步表明，这种策略可以成功地应用于运输到活细胞中的大蛋白质。我们的研究结果验证了一个有希望的第一步，在实现控制细胞渗透和使用CPP的治疗方法。
Cell-penetrating peptides are able to transport a wide variety of cargo across cell membranes. Although promising, they are not often considered for therapeutic purposes as they lack controllable activity and cell selectivity. We have developed an activation strategy based on a split octa-arginine cell-penetrating peptide (CPP) that can be activated by means of bioorthogonal ligation. To this end we prepared two non-penetrating tetra-arginine halves, functionalized either with a tetrazine or with a complementary bicyclo[6.1.0] nonyne (BCN) group. We demonstrate that an active octa-arginine can be reconstituted in situ upon mixing the complementary split peptides. The resulting activated peptide is taken up as efficiently as the well-established cell-penetrating peptide octa-arginine. The activation of the oligoarginines can also be achieved using trans-cyclooctene (TCO) as a ligation partner, while norbornene appears too kinetically slow for use in situ. We further show that this strategy can be applied successfully to transport a large protein into living cells. Our results validate a promising first step in achieving control over cell penetration and to use CPPs for therapeutic approaches.