Solid-phase synthesis of porcine cholecystokinin-33 in a new resin via FMOC-strategy.
Solid-phase synthesis of porcine cholecystokinin-33 in a new resin via FMOC-strategy.
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通过 FMOC 策略在新树脂中固相合成猪胆囊收缩素-33。
DOI：
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发表时间：
1991
期刊：
影响因子：
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通讯作者：
L. Nyerges
中科院分区：
文献类型：
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作者：
B. Penke;L. Nyerges
用固相合成法合成了猪缩胆囊素-33，并对其进行了化学和生物学表征。为了开发一种成功的合成策略，设计了一种新的锚分子(4-succinylamido-2，2‘，4’-trimethoxybenzhydrylamine)，并将其偶联到氨基甲基聚苯乙烯上。合成的4-succinylamido-2，2‘，4’-trimethoxybenzhydrylamine树脂被成功地用于以N-荧甲氧基甲氧基氨基酸对称酸酐为原料合成胆囊素-33。通过酪氨酸与氯磺酸的直接硫酸盐化反应合成了酪氨酸-O-硫酸酯，并以N-fluorenylmethoxy-O-sulfatotyrosine-pentafluorophenyl酯的形式偶联到多肽序列中。三官能团氨基酸的侧链主要由叔丁基型保护基团保护。精氨酸的胍基功能被2，2，5，7，8-五甲基色曼-6-磺酰基保护。合成完成后，用50%三氟乙酸(15min)将多肽从载体上切割下来，此处理同时切割侧链保护基团。制备性高效液相色谱分离得到具有完全生物学活性的纯CCK-33。通过氨基酸分析、红外光谱、紫外光谱、快原子轰击质谱仪以及合成的和天然的缩胆囊素-33的比较高效液相色谱鉴定了该多肽的结构。
Porcine cholecystokinin-33 has been synthesized on solid phase and characterized both chemically and biologically. In order to develop a successful synthetic strategy, a new anchor molecule (4-succinylamido-2,2',4'-trimethoxybenzhydrylamine) was designed and coupled to aminomethyl-polystyrene. The resulting 4-succinylamido-2,2',4'-trimethoxybenzhydrylamine resin was successfully used for the synthesis of cholecystokinin-33 using N-fluorenylmethoxycarbonyl amino acid symmetric anhydrides. Tyrosine-O-sulfate has been synthesized by direct sulfation of tyrosine with chlorosulfonic acid and incorporated into the peptide sequence by coupling as N-fluorenylmethoxy-O-sulfatotyrosine-pentafluorophenyl ester. Side chains of the trifunctional amino acids were protected mostly by t-butyl-type protecting groups. The guanidino function of arginine was protected by the 2,2,5,7,8-pentamethylchromane-6-sulfonyl group. After completion of the synthesis, the peptide was cleaved off the support with 50% trifluoroacetic acid (15 min); this treatment cleaved the side-chain protecting groups simultaneously. Preparative high-performance liquid chromatography resulted in pure cholecystokinin-33 of full biological activity. The structure of the peptide was proved by amino acid analysis, IR and UR spectroscopy, fast atomic bombardment mass spectroscopy and comparative high-performance liquid chromatography of the synthetic and native cholecystokinin-33.