Rapidly measuring reactivities of carboxylic acids to generate equireactive building block mixtures: a spectrometric assay.
The relative reactivity of building blocks is critical for a successful preparation of combinatorial libraries. Here, we present a method for measuring the reactivity of carboxylic acid building blocks in amide-forming reactions. The method involves competitive reactions between a reference and test acid and a tetraphenylporphyrin reaction partner with four reactive sites. Relative reactivities are calculated on the basis of the distribution of substituted porphyrins found in MALDI-TOF mass spectra. Reactivities thus determined were used to prepare reactivity-adjusted building block mixtures. These were reacted with amino-terminal oligonucleotide and peptide scaffolds on solid support, generating small libraries suitable for spectrometrically monitored selection experiments (SMOSE). The rate of building block "drop outs" that fail to couple as expected was not substantially lowered by acquiring spectra from two reactions, performed with different ratios of building blocks, where the effect of a given substituent on the desorption/ionization yield of the porphyrin can be eliminated. Instead, coupling building blocks of similar size together or employing N-hydroxysuccinimide esters rather than activating with a "uronium salt" were found to improve the quality of libraries generated via competitive reactions.
Dombi
Kendra L
KL
Department of Chemistry, University of Constance, 78457 Konstanz, Germany.
Steiner
Ulrich E
UE
Richert
Clemens
C
D000577:Amides / Q000138:chemical synthesis; D002264:Carboxylic Acids / Q000737:chemistry*; D020650:Combinatorial Chemistry Techniques; D009841:Oligonucleotides / Q000138:chemical synthesis; D010455:Peptides / Q000138:chemical synthesis; D011166:Porphyrins / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013057:Spectrum Analysis
D000577:Amides; D002264:Carboxylic Acids; D009841:Oligonucleotides; D010455:Peptides; D011166:Porphyrins; C409084:tetraphenylporphyrin