van Abel R J, Tang Y Q, Rao V S, Dobbs C H, Tran D, Barany G, Selsted M E
Department of Chemistry, University of Minnesota, Minneapolis, USA.
Int J Pept Protein Res. 1995 May;45(5):401-9. doi: 10.1111/j.1399-3011.1995.tb01055.x.
Indolicidin, a novel tryptophan-rich microbicidal tridecapeptide amide isolated originally from granules of bovine neutrophils, has been prepared by optimized manual and automated protocols of stepwise solid-phase synthesis with N alpha-9-fluorenylmethyloxycarbonyl (Fmoc) amino acid derivatives. Both standard polystyrene (PS) and polyethylene glycol-polystyrene (PEG-PS) graft supports were used in combination with handles that provide C-terminal peptide amides: 5-(4-Fmoc-aminomethyl-3,5-dimethoxyphenoxy)valeric acid (PAL) or 5-(9-Fmoc-aminoxanthen-2-oxy)valeric acid (XAL). Final deprotection/cleavage was carried out with reagent K, trifluoroacetic acid-phenol-water-thioanisole-1,2-ethanedithiol (82.5:5:5:5:2.5), or reagent B, trifluoroacetic acid-phenol-water-tri(isopropyl)silane (88:5:5:2), and related cocktails. Initial purities as high as 93% were obtained immediately following cleavage. In the largest-scale synthesis carried out, 0.8 g of HPLC-purified indolicidin (> 99% pure) was obtained, representing a 39% overall yield based on C-terminal Arg(Pmc) anchored to PAL-PS-resin. The main synthetic product, and some by-products, were characterized by analytical high-performance liquid chromatography (HPLC), sequencing, and fast atom bombardment mass spectrometry (FABMS). The antimicrobial potencies of natural and synthetic indolicidin, as determined by in vitro antibacterial and antifungal assays, were identical. Further, the reactivities of natural and synthetic peptides with anti-indolicidin antibody were indistinguishable.
吲哚杀菌素是一种最初从牛嗜中性粒细胞颗粒中分离出的富含色氨酸的新型杀微生物十三肽酰胺，已通过使用Nα-9-芴甲氧羰基（Fmoc）氨基酸衍生物的优化手动和自动逐步固相合成方案制备而成。标准聚苯乙烯（PS）和聚乙二醇-聚苯乙烯（PEG-PS）接枝载体均与提供C端肽酰胺的手柄结合使用：5-（4-Fmoc-氨基甲基-3,5-二甲氧基苯氧基）戊酸（PAL）或5-（9-Fmoc-氨基呫吨-2-氧基）戊酸（XAL）。最终脱保护/裂解使用试剂K、三氟乙酸-苯酚-水-硫代苯甲醚-1,2-乙二硫醇（82.5:5:5:5:2.5）或试剂B、三氟乙酸-苯酚-水-三（异丙基）硅烷（88:5:5:2）以及相关混合液进行。裂解后立即获得高达93%的初始纯度。在进行的最大规模合成中，获得了0.8 g经高效液相色谱（HPLC）纯化的吲哚杀菌素（纯度>99%），基于锚定在PAL-PS树脂上的C端精氨酸（Pmc），总产率为39%。主要合成产物和一些副产物通过分析型高效液相色谱（HPLC）、测序和快原子轰击质谱（FABMS）进行了表征。通过体外抗菌和抗真菌试验测定，天然和合成吲哚杀菌素的抗菌效力相同。此外，天然和合成肽与抗吲哚杀菌素抗体的反应性无法区分。
