RESUMO
Drugs are designed and validated based on physicochemical data on their interactions with target proteins. For low water-solubility drugs, however, quantitative analysis is practically impossible without accurate estimation of precipitation. Here we combined quantitative NMR with NMR titration experiments to rigorously quantify the interaction of the low water-solubility drug pimecrolimus with its target protein FKBP12. Notably, the dissociation constants estimated with and without consideration of precipitation differed by more than tenfold. Moreover, the method enabled us to quantitate the FKBP12-pimecrolimus interaction even under a crowded condition established using the protein crowder BSA. Notably, the FKBP12-pimecrolimus interaction was slightly hampered under the crowded environment, which is explained by transient association of BSA with the drug molecules. Collectively, the described method will contribute to both quantifying the binding properties of low water-solubility drugs and to elucidating the drug behavior in complex crowded solutions including living cells.
Assuntos
Soroalbumina Bovina/química , Proteína 1A de Ligação a Tacrolimo/química , Tacrolimo/análogos & derivados , Animais , Bovinos , Espectroscopia de Ressonância Magnética , Solubilidade , Tacrolimo/química , Água/químicaRESUMO
Although enantiomeric separation of single-wall carbon nanotubes is possible, their enantiomeric purity (EP) remains an issue due to a lack of effective evaluation methods. In this work, we report the EP of (6,5) carbon nanotube enantiomers using flavin mononucleotide (FMN) as an enantiomer-sensitive dispersant. The enantiomers (6,5) and (11,-5) were separated by a gel column chromatography method and dispersed in a FMN aqueous solution. In these solutions, (6,5) and (11,-5) showed E11 optical transitions at different wavelengths due to handedness-dependent interactions with the FMN molecule, which enabled us to estimate each concentration, namely, the EP. We prepared six intermediate-purity enantiomer samples by mixing the (6,5) and (11,-5) enantiomers and measured their circular dichroism (CD) spectra. The CD signal was confirmed to change linearly with the EP. Using this relationship, we can estimate the EP of any mixture of (6,5) and (11,-5) from its CD intensity.
Assuntos
Mononucleotídeo de Flavina/química , Nanotubos de Carbono/química , Cromatografia Líquida de Alta Pressão , Dicroísmo Circular , Nanotecnologia , EstereoisomerismoRESUMO
We report the chirality and enantiomer separation of metallic single-wall carbon nanotubes (SWCNTs) using gel chromatography, which has been the last remaining issue in SWCNT separation that has yet to be achieved. The key to the separation is summarized as the following three points: (i) the use of a preseparated metallic SWCNT mixture to eliminate the semiconducting SWCNTs that are more interactive with the gel; (ii) the reduction of the concentration of dispersant to increase the interaction between the metallic SWCNTs and the gel; and (iii) the use of a long column to increase the number of interaction sites that enhance the slight differences between metallic SWCNT species. Using these three separation conditions, we obtained chirality-sorted metallic SWCNTs, especially (10,4) metallic SWCNTs were highly enriched. Circular dichroism spectra demonstrated the enantiomer separation of metallic SWCNTs. The discrimination of the enantiomers is derived from the dextran in the gel, which is the only enantiomeric moiety in this system. This is the first report on the enantiomer separation of metallic SWCNTs and will contribute to progress in the fundamental physics and applications of SWCNTs.
RESUMO
The palladium-catalyzed regio- and enantioselective allylic amination of trifluoromethyl group substituted racemic and unsymmetrical 1,3-disubstituted allylic esters has been accomplished. The enantioselective formation of the α-type allylic amines was attained by the dynamic kinetic asymmetric transformation (DYKAT).
RESUMO
It found that the Ru(3)(CO)(12) coordinated with 2-(diphenylphosphino)benzonitrile (2-DPPBN) to effectively catalyze the [2 + 2 + 2] cyclotrimerization of the trifluoromethyl group substituted internal alkynes in high yields with up to >98% regioselectivity. Isolation of a ruthenacyclopentadiene was successful and confirmed that the complex is a reaction intermediate.