The 'O-acyl isopeptide method' for the synthesis of difficult sequence-containing peptides: application to the synthesis of Alzheimer's disease-related amyloid beta peptide (Abeta) 1-42.
An efficient 'O-acyl isopeptide method' for the synthesis of difficult sequence-containing peptides was applied successfully to the synthesis of amyloid beta peptide (Abeta) 1-42 via a water-soluble O-acyl isopeptide of Abeta1-42, i.e. '26-O-acyl isoAbeta1-42' (6). This paper describes the detailed synthesis of Abeta1-42 focusing on the importance of resin selection and the analysis of side reactions in the O-acyl isopeptide method. Protected '26-O-acyl isoAbeta1-42' peptide resin was synthesized using 2-chlorotrityl chloride resin with minimum side reactions in comparison with other resins and deprotected crude 26-O-acyl isoAbeta1-42 was easily purified by HPLC due to its relatively good purity and narrow elution with reasonable water solubility. This suggests that only one insertion of the isopeptide structure into the sequence of the 42-residue peptide can suppress the unfavourable nature of its difficult sequence. The migration of O-acyl isopeptide to intact Abeta1-42 under physiological conditions (pH 7.4) via O--N intramolecular acyl migration reaction was very rapid and no other by-product formation was observed while 6 was stable under storage conditions. These results concluded that our strategy not only overcomes the solubility problem in the synthesis of Abeta1-42 and can provide intact Abeta1-42 efficiently, but is also applicable in the synthesis of large difficult sequence-containing peptides at least up to 50 amino acids. This synthesis method would provide a biological evaluation system in Alzheimer's disease research, in which 26-O-acyl isoAbeta1-42 can be stored in a solubilized form before use and then rapidly produces intact Abeta1-42 in situ during biological experiments.
Sohma
Youhei
Y
Department of Medicinal Chemistry, Center for Frontier Research in Medicinal Science, 21st Century COE Program, Kyoto Pharmaceutical University, Yamashina-Ku, Japan.
Hayashi
Yoshio
Y
Kimura
Maiko
M
Chiyomori
Yousuke
Y
Taniguchi
Atsuhiko
A
Sasaki
Masato
M
Kimura
Tooru
T
Kiso
Yoshiaki
Y
D000215:Acylation; D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D004121:Dimethyl Sulfoxide; D004355:Drug Stability; D000432:Methanol; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry; D011137:Polystyrenes; D012694:Serine / Q000737:chemistry; D012995:Solubility; D014867:Water
D016229:Amyloid beta-Peptides; D002021:Buffers; D010446:Peptide Fragments; D011137:Polystyrenes; C075222:amyloid beta-protein (1-42); D014867:Water; C101297:Tentagel resin; D012694:Serine; D000432:Methanol; D004121:Dimethyl Sulfoxide