RESUMEN
For decades now, the separation of chiral enantiomers of drugs has been gaining the interest and attention of researchers. In 1991, the first guidelines for development of chiral drugs were firstly released by the US-FDA. Since then, the development in chromatographic enantioseparation tools has been fast and variable, aiming at creating a suitable environment where the physically and chemically identical enantiomers can be separated. Among those tools, the immobilization of chiral selectors (CS) on different stationary phases and the chiral mobile phase additives (CMPA) which have been progressed and studied extensively. This review article highlights the major advances in immobilization of CS together with their different recognition mechanisms as well as CMPA as a cheaper and successful alternative for chiral stationary phases. Moreover, the role of molecular modeling tool as a pre-step in the choice of CS for evaluating possible interactions with different ligands has been pointed up. Illustrations of reported methods and updates for immobilized CS and CMPA have been included.
Asunto(s)
Cromatografía Líquida de Alta Presión , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Liquida , Estereoisomerismo , LigandosRESUMEN
Terbutaline is mainly metabolized by sulfoconjugation stereoselectively, favoring its (S)-(+) enantiomer. Reported chiral separations of Terbutaline enantiomers were achieved by various chromatographic methods. However, the simultaneous enantioseparation of Terbutaline and the monosulfate conjugate metabolites was never reported. This study aims at shedding light on the influential factors and interactions leading to successful enantioseparation of Terbutaline and its monosulfate conjugate pairs by Supercritical Fluid Chromatography (SFC) for the first time within a Quality by Design framework using Design of Experiments. The effect of molarity of mobile phase additive, mobile phase flow rate, column temperature and back pressure were evaluated. Compared to previous reports, the response surface interestingly revealed the favorability of high temperature and high flow rate up to 2.25 ml/min for resolution of the two pairs of enantiomers on polysaccharide chiral stationary phase CHIRALPAK IC. In addition, a switch in the elution order of Terbutaline and the sulfate conjugate peak pairs was observed upon elimination of the mobile phase additive where the sulfate conjugate underwent intra-molecular ionic interactions and the change in elution order was only due to TER behavior. The multifactorial interactions would not have been detected with the common one-factor-at-a-time approach during method development, demonstrating the superiority and importance of the Analytical Quality by Design frame in enantioseparation.
Asunto(s)
Cromatografía con Fluido Supercrítico , Cromatografía con Fluido Supercrítico/métodos , Polisacáridos/química , Estereoisomerismo , Sulfatos , TerbutalinaRESUMEN
The emergence of bacterial resistance has triggered a multitude of efforts to develop new antibacterial agents. There are many compounds in literature that were reported as potent antibacterial agents, however, they lacked the required safety to mammalian cells or no clear picture about their toxicity profile was presented. Inspired by discovered hit from our in-house library and by previously reported 2,4-diaminosubstituted quinazolines, we describe the design and synthesis of novel 2,4-disubstituted-thioquinazolines (3-13 and 36), 2-thio-4-amino substituted quinazolines (14-33) and 6-substituted 2,4-diamonsubstituted quinazolines (37-39). The synthesized compounds showed potent antibacterial activity against a panel of Gram-positive, efflux deficient E.coli and Mycobacterium smegmatis. The panel also involved resistant strains including methicillin-resistant Staphylococcus aureus, penicillin-resistant Streptococcus pneumoniae, vancomycin-resistant Enterococcus faecalis and vancomycin-resistant Enterococcus faecium, in addition to Mycobacterium smegmatis. The newly synthesized compounds revealed MIC values against the tested strains ranging from 1 to 64 µg/mL with a good safety profile. Most of the 2-thio-4-amino substituted-quinazolines showed significant antimycobacterial activity with the variations at position 2 and 4 offering additional antibacterial activity against the different strains. Compared to previously reported 2,4-diaminosubstituted quinazolines, the bioisosteric replacement of the 2-amino with sulfur offered a successful approach to keep the high antibacterial potency while substantially improving safety profile as indicated by the reduced activity on different cell lines and a lack of hemolytic activity.
Asunto(s)
Antibacterianos/farmacología , Quinazolinas/farmacología , Animales , Antibacterianos/síntesis química , Antibacterianos/química , Células CHO , Supervivencia Celular/efectos de los fármacos , Cricetulus , Relación Dosis-Respuesta a Droga , Enterococcus faecalis/efectos de los fármacos , Células Hep G2 , Humanos , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Mycobacterium smegmatis/efectos de los fármacos , Quinazolinas/síntesis química , Quinazolinas/química , Staphylococcus aureus/efectos de los fármacos , Streptococcus pneumoniae/efectos de los fármacos , Relación Estructura-ActividadRESUMEN
The design of experiments (DoE) is one of the quality-by-design tools valued in analytical method development, not only for cost reduction and time effectiveness, but also for enabling analytical method control and understanding via a systematic workflow, leading to analytical methods with built-in quality. This work aimed at using DoE to enhance method understanding for a developed UHPLC enantioseparation of terbutaline (TER), a model chiral drug, and to define quality assurance parameters associated with using chiral mobile phase additives (CMPA). Within a response surface methodology workflow, the effect of different factors on both chiral resolution and retention was screened and optimized using Plackett-Burman and central composite designs, respectively, followed by multivariate mathematical modeling. This study was able to delimit method robustness and elucidate enantiorecognition mechanisms involved in interactions of TER with the chiral modifiers. Among many CMPAs, successful TER enantioresolution was achieved using hydroxypropyl ß-cyclodextrin (HP-ß-CD) added to the mobile phase as 5.4 mM HP-ß-CD in 52.25 mM ammonium acetate. Yet, limited method robustness was observed upon switching between the different tested CMPA, concluding that quality can only be assured with specific minimal pre-run conditioning time with the CMPA, namely 16-column volume (60 min at 0.1 mL/min). For enantiorecognition understanding, computational molecular modeling revealed hydrogen bonding as the main binding interaction, in addition to dipole-dipole inside the CD cavity for the R enantiomer, while the S enantiomer was less interactive.