Cleavage kinetics and anchor linked intermediates in solid phase peptide amide synthesis.
Kinetics and cleavage conditions of peptide amide synthesis were studied using the anchor molecules 5-(4'-aminomethyl-3',5'-dimethoxyphenoxy)valeric acid (4-ADPV-OH) and 5-(2'-aminomethyl-3'-5'-dimethoxyphenoxy) valeric acid (2-ADPV-OH). Unexpectedly the anchor amide alanyl-4-ADPV-NH2 was isolated and characterized as an intermediate during the cleavage with trifluoroacetic acid (TFA) of alanyl-4-ADPV-alanyl-aminomethyl-polystyrene to yield the alanine amide. As a matter of fact the NH--CH alpha bond of the alanyl spacer has to be cleaved to form this intermediate. Using TFA-dichloromethane (1:9) alanyl-4-ADPV-NH2 was obtained as a cleavage product in 50% yield within 60 min, whereas the isomeric alanyl-2-ADPV-NH2 was formed more slowly under these mild conditions. At high TFA concentration no difference between the 2- and 4-ADPV anchor was observed in the rate of formation of the free alanine amide. The presence of tryptophan amide in the cleavage mixture resulted in an anchor alkylated tryptophan amide, which remains stable in acidic solution but disappears rapidly in the presence of the resin. A low TFA/high TFA cleavage procedure is recommended for peptide amid synthesis applying the ADPV anchor.
Dürr
H
H
Institute for Organic Chemistry, University of Tübingen, FRG.
Beck-Sickinger
A G
AG
Schnorrenberg
G
G
Rapp
W
W
Jung
G
G
D000478:Alkylation; D000577:Amides / Q000737:chemistry; D005449:Fluorenes / Q000737:chemistry*; D007700:Kinetics; D013058:Mass Spectrometry; D010455:Peptides / Q000138:chemical synthesis*; D012117:Resins, Synthetic / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D014269:Trifluoroacetic Acid / Q000737:chemistry; D014631:Valerates / Q000737:chemistry*
C072877:5-(2-(9-fluorenylmethyloxycarbonyl)aminomethyl-3,5-dimethoxyphenoxy)valeric acid; D000577:Amides; D005449:Fluorenes; D010455:Peptides; D012117:Resins, Synthetic; D014631:Valerates; C064395:5-(4-(9-fluorenylmethyloxycarbonyl)aminomethyl-3,5-dimethoxyphenoxy)valeric acid; D014269:Trifluoroacetic Acid