Deprotonation and regioselective addition of 2H-pyrazolo[3,4-c]quinolines to electrophiles.

The deprotonation and regioselective reaction of 2H-pyrazolo[3,4-c]quinolines with a variety of electrophiles is described. Electrophiles include benzaldehyde, DMF, carbon dioxide, and iodine. This method provides a direct route to a class of pharmacologically interesting compounds.

A kinetic study of product cleavage reactions from the solid phase by a biocompatible and removable cleaving reagent, HCl.

TFA has been widely used as a cleaving reagent in solid-phase organic synthesis. However, it is difficult to remove from the final product, and it is toxic to various cells. To search for an alternative, we studied the kinetics of HCl cleavage reactions of 18 resin-bound compounds on various linkers. HCl is very easy to remove completely from samples, and the residual HCl does not have a toxic effect in cell assays. Most compounds studied in this work can be easily cleaved using a low concentration of HCl (0.9-2.3%) and the minimal amount of time (60-90 min). Even in the most difficult case, a moderate 8% HCl and an extended time (10-15 h) are enough to cleave the product. Therefore, our kinetic studies establish HCl as a biocompatible, removable, and effective substitute for TFA when final compounds are used for biological screening and drug discovery.

Quality control in combinatorial chemistry: determination of the quantity, purity, and quantitative purity of compounds in combinatorial libraries.

The quality of combinatorial libraries determines the success of biological screening in drug discovery programs. In this paper, we evaluate and compare various methods for measuring identity, purity, and quantity (yield) of combinatorial libraries. Determination of quantitative purity reveals the true library quality and often indicates potential quality problems before full-scale library production. The relative purity can be determined for every member in a large library in a high-throughput mode, but must be cautiously interpreted. In particular, many impurities are not observable by relative purity measurements using detectors such as UV(214), UV(254), and evaporative light-scattering detection. These "invisible" impurities may constitute a significant portion of the sample weight. We found that TFA, plastic extracts, inorganic compounds, and resin washout are among these impurities. With compelling evidence, we reach a conclusion that purification is the only way to remove "invisible" impurities and improve the quantitative purity of any compound even though some compounds may have a high relative purity before purification.

Kinetics study of amine cleavage reactions of various resin-bound thiophenol esters from marshall linker.

The kinetics of cleavage reactions of seven resin-bound thiophenol esters with three amines has been studied by single-bead FTIR. The reactivity of these seven thiophenol esters was dependent on their structures and could be summarized as follows: 5-benzimidazolecarboxylic thiophenol ester > alkyl thiophenol ester > aromatic thiophenol ester. The reactivity of three amines was summarized as follows: n-butylamine > 3,4-dimethoxyphenethylamine > 1-piperonylpiperazine. The rate of the cleavage reaction increased 2-fold per 10 degrees C rise in reaction temperature. Oxidation of the thiophenol linker increased the rate of the cleavage reaction by 580-fold.

Color test for the detection of resin-bound aldehyde in solid-phase combinatorial synthesis.

We report the development of a sensitive and specific color test for the detection of the presence of resin-bound aldehyde groups using 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole (Purpald). Aldehyde resin turns dark-brown to purple after a 5 min reaction followed by a 10 min air oxidation period. Resins that possess other functional groups (i.e., ketone, ester, amide, alcohol, and carboxylic acid) do not change color under the same conditions. The detection limit is 20 micromol/g for polystyrene-based aldehyde resins.