Cytotoxicity and vitreous stability of chemically modified connexin43 mimetic peptides for the treatment of optic neuropathy
Cytotoxicity and vitreous stability of chemically modified connexin43 mimetic peptides for the treatment of optic neuropathy
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DOI：
10.1002/jps.23617
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发表时间：
2013-07-01
影响因子：
3.8
通讯作者：
Rupenthal, Ilva D.
中科院分区：
文献类型：
--
作者：
Chen, Ying-Shan;Toth, Istvan;Rupenthal, Ilva D.
视神经病变与视网膜神经节细胞（RGC）丢失有关，导致视神经损伤和视力障碍。无调节连接蛋白（Cx）半通道开放在RGC丢失中起作用。因此，通过cx43特异性模拟肽（MP）抑制可以防止进一步的细胞死亡。然而，天然多肽的高亲水性和较差的稳定性阻碍了它们在生物膜上的有效传递。本研究旨在通过偶联c12 -脂氨基酸（C12-Laa）或糖基来提高Cx43 MP的稳定性。采用固相肽合成法合成未修饰和修饰的Cx43 MP。它们的功能是通过碘化丙烯（PI）摄取到NT2细胞（一种能够分化为星形胶质细胞的人睾丸癌祖细胞系）来评估的，而在眼玻璃体中的稳定性是通过反相高效液相色谱法测量的。PI摄取研究显示未修饰和修饰的Cx43 MP对半通道开放有抑制作用。稳定性测量表明，改进的Cx43 MP的稳定性得到了改善，两个Laa基团使牛玻璃体中的肽半衰期增加了两倍以上。与C12-Laa或糖偶联不影响Cx43 MP的功能，但添加两个C12-Laa基团可显著提高肽的稳定性。因此，laa修饰可以改善多肽在体内的稳定性和视网膜递送。(3)中国医药科学，2013 (2):1 - 3
Optic neuropathy is associated with retinal ganglion cell (RGC) loss leading to optic nerve damage and visual impairment. Unregulated connexin (Cx) hemichannel opening plays a role in RGC loss. Thus, inhibition via Cx43-specific mimetic peptides (MP) may prevent further cell death. However, the highly hydrophilic character and poor stability of native peptides prevent their efficient delivery across biological membranes. The present study aimed to improve the stability of Cx43 MP by conjugation to C12-lipoamino acid (C12-Laa) or sugar groups. Unmodified and modified Cx43 MP were synthesized using solid-phase peptide synthesis. Their functionality was assessed by propidium iodide (PI) uptake into NT2 cells, a human testicular carcinoma progenitor cell line able to differentiate into astrocytes, whereas the stability in ocular vitreous was measured by reversed-phase high-performance liquid chromatography. PI uptake studies showed inhibition of hemichannel opening for unmodified and modified Cx43 MP. Stability measurements revealed improved stability of modified Cx43 MP, with two Laa groups increasing the peptide half-life in bovine vitreous more than twofold. Conjugation to C12-Laa or sugar did not affect the functionality of Cx43 MP, but addition of two C12-Laa groups significantly improved peptide stability. Laa-modifications may therefore offer improved stability and retinal delivery of peptides in vivo. (c) 2013 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:2322-2331, 2013