RESUMO
Structural mass spectrometry (MS) techniques are fast and sensitive analytical methods to identify noncovalent guest/host complexation phenomena for desirable solution-phase properties. Current MS-based studies on guest/host complexes of drug and drug-like molecules are sparse, and there is limited guidance on how to interpret MS information in the context of host nanoencapsulation and inclusion. Here, we use structural MS strategies, combining energy-resolved MS (ERMS), ion mobility-MS (IM-MS), and computational modeling, to characterize 14 chemically distinct drug and drug-like compounds for their propensity to form guest/host complexes with the widely used excipient, beta-cyclodextrin (ßCD). The majority (11/14) yielded a 1:1 guest/host complex, and ion mobility collision cross section (CCS) analysis provided subtle evidence of gas-phase compaction of complexes in both polarities. The three distinct dissociation channels observed in ERMS (i.e., charged ßCD, charged guest, and partial guest loss) were used to direct charge-site assignments for computational modeling, and structural candidates were prioritized using helium-derived CCS measurements combined with root-mean-square distance analysis. The combined analytical information from ERMS, IM-MS, and computational modeling suggested that the majority of anhydrous complexes are inclusion complexes with ßCD. Taken together, this work demonstrates a roadmap for how multiple MS-based analytical measurements can be combined to interpret the structures that guest/host complexes adopt in the absence of water.
RESUMO
Drug enantiomers can possess vastly different pharmacological properties, yet they are identical in their chemical composition and structural connectivity. Thus, resolving enantiomers poses a great challenge in the field of separation science. Enantiomer separations necessitate interaction of the analyte with a chiral environmentâin mass spectrometry-based analysis, a common approach is through a three-point interaction with a chiral selector commonly introduced during sample preparation. In select cases, the structural difference imparted through noncovalent complexation results in enantiomer-specific structural differences, facilitating measurement using a structurally selective analytical technique such as ion mobility-mass spectrometry (IM-MS). In this work, we investigate the direct IM-MS differentiation of chiral drug compounds using mononuclear copper complexes incorporating an amino acid chiral selector. A panel of 20 chiral drugs and drug-like compounds were investigated for separation, and four l-amino acids (l-histidine, l-tryptophan, l-proline, and l-tyrosine) were evaluated as chiral selectors (CS) to provide the chiral environment necessary for differentiation. Enantiomer differentiation was achieved for four chiral molecule pairs (R/S-thalidomide, R/S-baclofen, R/S-metoprolol, and d/l-panthenol) with two-peak resolution (Rp-p) values ranging from 0.7 (>10% valley) to 1.5 (baseline separation). Calibration curves relating IM peak areas to enantiomeric concentrations enabled enantiomeric excess quantitation of racemic thalidomide and metoprolol with residuals of 5.7 and 2.5%, respectively. Theoretical models suggest that CuII and l-histidine complexation around the analyte chiral center is important for gas-phase stereoselectivity. This study demonstrates the potential of combining enantioselective noncovalent copper complexation with structurally selective IM-MS for differentiating chiral drugs and drug-like compounds.
Assuntos
Aminoácidos , Cobre , Espectrometria de Mobilidade Iônica , Cobre/química , Estereoisomerismo , Aminoácidos/química , Aminoácidos/análise , Espectrometria de Mobilidade Iônica/métodos , Espectrometria de Massas/métodos , Preparações Farmacêuticas/química , Preparações Farmacêuticas/análise , Complexos de Coordenação/química , Estrutura MolecularRESUMO
Cyclodextrins (CDs) are a family of macrocyclic oligosaccharides with amphiphilic properties, which can improve the stability, solubility, and bioavailability of therapeutic compounds. There has been growing interest in the advancement of efficient and reliable analytical methods that assist with elucidating CD host-guest drug complexation. In this study, we investigate the noncovalent ion complexes formed between naturally occurring dextrins (αCD, ßCD, γCD, and maltohexaose) with the poorly water-soluble antimalarial drug, artemisinin, using a combination of ion mobility-mass spectrometry (IM-MS), tandem MS/MS, and theoretical modeling approaches. This study aims to determine if the drug can complex within the core dextrin cavity forming an inclusion complex or nonspecifically bind to the periphery of the dextrins. We explore the use of group I alkali earth metal additives to promote the formation of various noncovalent gas-phase ion complexes with different drug/dextrin stoichiometries (1:1, 1:2, 1:3, 1:4, and 2:1). Broad IM-MS collision cross section (CCS) mapping (n > 300) and power-law regression analysis were used to confirm the stoichiometric assignments. The 1:1 drug:αCD and drug:ßCD complexes exhibited strong preferences for Li+ and Na+ charge carriers, whereas drug:γCD complexes preferred forming adducts with the larger alkali metals, K+, Rb+, and Cs+. Although the ion-measured CCS increased with cation size for the unbound artemisinin and CDs, the 1:1 drug:dextrin complexes exhibit near-identical CCS values regardless of the cation, suggesting these are inclusion complexes. Tandem MS/MS survival yield curves of the [artemisinin:ßCD + X]+ ion (X = H, Li, Na, K) showed a decreased stability of the ion complex with increasing cation size. Empirical CCS measurements of the [artemisinin:ßCD + Li]+ ion correlated with predicted CCS values from the low-energy theoretical structures of the drug incorporated within the ßCD cavity, providing further evidence that gas-phase inclusion complexes are formed in these experiments. Taken together, this work demonstrates the utility of combining analytical information from IM-MS, MS/MS, and computational approaches in interpreting the presence of gas-phase inclusion phenomena.
Assuntos
Artemisininas , Ciclodextrinas , Dextrinas , Espectrometria de Massas em Tandem , Ciclodextrinas/química , Cátions/químicaRESUMO
Previous work has demonstrated that copper complexation strategies can be used with tandem MS (MS/MS) and, more recently, ion mobility-mass spectrometry (IM-MS) to differentiate chiral isomers based upon enantiomeric-specific binding. In this study, we investigate the separation of chiral amino acids (AAs) forming trinuclear complexes that can be directly resolved by IM-MS analyses. Twenty standard AAs of both d- and l-chirality were investigated. Specific AAs including d/l-histidine, d/l-proline, d/l-glutamine, d/l-tyrosine, and d/l-tryptophan were evaluated as "chiral selectors" that, when combined with copper, were found to promote selective complexation with specific AA enantiomers. Significant enantiomer differentiation was observed in the IM spectra for hydrophobic AAs acids with peak-to-peak resolutions ranging from 0.63 to 1.15. Among the chiral selectors investigated, histidine provided the best enantioselectivity, followed by tryptophan, suggesting the aromatic structure plays an important role in forming chiral-specific ion complexes. Unlike MS/MS methods where chiral selectors with l-stereochemistry enhance the differentiation, the chirality of the selector was found to have no significant effect on observed IM separation with both d- and l-selectors providing similar resolutions but with inverted IM arrival time ordering. To investigate the structural differences between resolvable chiral complexes, a combination of MS/MS, collision cross-section (CCS) measurements, and molecular mechanics techniques was used. Candidate trinuclear structures of the stoichiometry [(Cu2+)3(d/lIle)3(lHis)2 - 5H]+ were constructed with guidance from empirical MS/MS results. Of the 48 theoretical structures generated, one enantiomeric cluster pair yielded close correlation (<1%) with experimental CCS measurements, suggesting the most enantioselective ion complexes observed in this work are bridged by three coppers.
Assuntos
Aminoácidos , Cobre , Aminoácidos/química , Cobre/química , Histidina/química , Espectrometria de Mobilidade Iônica , Estereoisomerismo , Espectrometria de Massas em Tandem/métodos , TriptofanoRESUMO
RATIONALE: Commercial-grade polymer synthesis is performed via melt polymerization, which leads to polydispersion. The work reported herein provides a synthetic strategy to produce mono-dispersive polyurethane oligomers and an analytical strategy to distinguish these oligomers, providing chemists with the tools necessary to synthesize and identify specific polymer structures that exhibit a desired property. METHODS: Three isomeric poly(ethylene glycol)-polyurethane (PEG-PUR) oligomers were synthesized and analyzed via flow-injection ion mobility mass spectrometry (IM-MS). Each polymer oligomer was injected and run independently via flow injection at 100 µLâ¢min-1 and analyzed in positive ion mode on a drift tube quadrupole time-of-flight (QTOF) instrument. Mobility measurements were determined using a single-field approach. For tandem mass spectrometry (MS/MS) experiments, the sodium-adducted singly charged precursor ion was isolated in the quadrupole and subjected to a range of collision energies. RESULTS: In MS experiments, both +1 and +2 sodium-adducted species were observed for each oligomer at m/z 837.4 and 430.2, respectively. When isolated and fragmented via MS/MS, the +1 precursor yielded distinct product ions for each of the three isomeric oligomers. Fragmentation generally occurred at urethane linkages via 1,3- and 1,5-H shift mechanisms. IM was also used to distinguish the three isomers, with greater IM separation observed for the +2 versus the +1 species. CONCLUSIONS: Mono-disperse PEG-PUR oligomers were synthesized and analyzed. Although the polymeric oligomers analyzed in this study are quite small and structurally simple, this work serves as a model system for the synthesis and structural characterization of larger, more complex block copolymers.