RESUMO
A new strategy for the preparation of distinct N-substituted muropeptides is described. Different orthogonally N-protected disaccharide thioglycosides were designed and synthesized. Among them, compound 4, qualified as a key intermediate, was utilized for further chemical transformations to develop a series of diverse N-substituted-glucosaminyl N-substituted-muramyl dipeptides (GMDPs). These unique muropeptides were applied for the study of human NOD2 stimulation. Intriguingly, structural modification of the MurNAc residue to N-non-substituted muramic acid (MurNH2 ) in GMDP dramatically impaired NOD2 stimulatory activity, but GMDPs possessing the glucosamine residue with a free amino group retained NOD2 stimulation activity. This work is the first study to illustrate the impact of both N-substituents of GMDPs on immunostimulatory activities of human NOD2.
Assuntos
Dipeptídeos , Dissacarídeos , Humanos , Proteína Adaptadora de Sinalização NOD2/metabolismoRESUMO
The imaging of peptidoglycan (PGN) dynamics in living bacteria facilitates the understanding of PGN biosynthesis and wall-targeting antibiotics. The main tools for imaging bacterial PGN are fluorescent probes, such as the well-known PGN metabolic labeling probes. However, fluorescent small-molecule probes for labeling key PGN-synthesizing enzymes, especially for transglycosylases (TGases), remain to be explored. In this work, the first imaging probe for labeling TGase in bacterial cell wall studies is reported. We synthesized various fluorescent MoeA-based molecules by derivatizing the natural antibiotic moenomycin A (MoeA), and used them to label TGases in living bacteria, monitor bacterial growth and division cycles by time-lapse imaging, and study cell wall growth in the mecA-carrying methicillin-resistant Staphylococcus aureus (MRSA) strains when the ß-lactam-based probes were unsuitable.
Assuntos
Antibacterianos/farmacologia , Bambermicinas/farmacologia , Parede Celular/efeitos dos fármacos , Corantes Fluorescentes/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Imagem Óptica , Peptidoglicano/análise , Antibacterianos/química , Bambermicinas/química , Parede Celular/metabolismo , Corantes Fluorescentes/química , Staphylococcus aureus Resistente à Meticilina/metabolismo , Testes de Sensibilidade Microbiana , Estrutura Molecular , Peptidoglicano/biossínteseRESUMO
A series of muramyl dipeptide (MDP) analogues with structural modifications at the C4 position of MurNAc and on the d-iso-glutamine (isoGln) residue of the peptide part were synthesized. The C4-diversification of MurNAc was conveniently achieved by using CuAAC click strategy to conjugate an azido muramyl dipeptide precursor with structurally diverse alkynes. d-Glutamic acid (Glu), replaced with isoGln, was applied for the structural diversity through esterification or amidation of the carboxylic acid. In total, 26â MDP analogues were synthesized and bio-evaluated for the study of human NOD2 stimulation activity in the innate immune response. Interestingly, MDP derivatives with an ester moiety are found to be more potent than reference compound MDP itself or MDP analogues containing an amide moiety. Among the varied lengths of the alkyl chain in ester derivatives, the MDP analogue bearing the d-glutamate dodecyl (C12) ester moiety showed the best NOD2 stimulation potency.
Assuntos
Acetilmuramil-Alanil-Isoglutamina/farmacologia , Proteína Adaptadora de Sinalização NOD2/metabolismo , Acetilmuramil-Alanil-Isoglutamina/análogos & derivados , Acetilmuramil-Alanil-Isoglutamina/química , Linhagem Celular , Humanos , Modelos Moleculares , Conformação MolecularRESUMO
Lipid II analogues bearing major modifications on the second sugar (GlcNAc) were synthesized and evaluated for their substrate activity toward TGases. Unexpectedly, N-deacetyled lipid II decreased its activity dramatically, and the C4-axial OH lipid II became an inhibitor (IC50 = 8 µM) with an approximately 14-fold increase in binding affinity toward TGase (25 vs. 27).
Assuntos
Clostridioides difficile/enzimologia , Inibidores Enzimáticos/farmacologia , Escherichia coli/enzimologia , Lipídeos/farmacologia , Peptidoglicano Glicosiltransferase/antagonistas & inibidores , Açúcares/farmacologia , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Lipídeos/química , Peptidoglicano Glicosiltransferase/metabolismo , Relação Estrutura-Atividade , Especificidade por Substrato , Açúcares/síntese química , Açúcares/químicaRESUMO
A convenient, efficient protocol to prepare diverse spiroisoxazolino-diketopiperazines via a parallel solid-supported synthesis was developed. The key steps are (1) a coupling reaction of an amino acid; (2) tosylation with concomitant ß-elimination to form an α, ß-unsaturated ester; (3) a 1,3-dipolar cycloaddition with an oxime to form isoxazoline rings; and (4) cyclic cleavage to release the product from the resin. All reaction steps and workup procedures were modified to allow the use of automated or semiautomated equipment. A 100-member demonstration library with two diversity sites was prepared in good purity and acceptable overall yields.
Assuntos
Piperazinas/síntese química , Técnicas de Síntese em Fase Sólida , Modelos QuímicosRESUMO
The preparation of natural product-like polyhydroxylated pyrrolidine and piperidine alkaloids using a combination of solid- and solution-phase organic synthesis is described. The key intermediates, enantiopure five- or six-membered tri-O-benzyl cyclic nitrones, were efficiently prepared on solid support from accessible chiral furanosides and pyranosides, respectively. The substituent diversity was achieved by a diastereoselective addition of a variety of Grignard reagents to the cyclic nitrones in solution-phase synthesis. All reaction steps and work-up procedures were modified to allow the use of automated equipment. A 36-membered demonstration library with three diversity elements (core, configuration, and substituent) was prepared in good yield and purity.
Assuntos
Alcaloides/química , Alcaloides/síntese química , Produtos Biológicos/química , Piperidinas/química , Pirrolidinas/química , Hidroxilação , Óxidos de Nitrogênio/síntese química , Óxidos de Nitrogênio/química , Bibliotecas de Moléculas Pequenas/síntese química , Bibliotecas de Moléculas Pequenas/química , Soluções , VolatilizaçãoRESUMO
To identify new transglycosylase inhibitors with potent anti-methicillin-resistant Staphylococcus aureus (MRSA) activities, a high-throughput screening against Staphylococcus aureus was conducted to look for antibacterial cores in our 2M compound library that consists of natural products, proprietary collection, and synthetic molecules. About 3600 hits were identified from the primary screening and the subsequent confirmation resulted in a total of 252 compounds in 84 clusters which showed anti-MRSA activities with MIC values as low as 0.1 µg/ml. Subsequent screening targeting bacterial transglycosylase identified a salicylanilide-based core that inhibited the lipid II polymerization and the moenomycin-binding activities of transglycosylase. Among the collected analogues, potent inhibitors with the IC(50) values below 10 µM against transglycosylase were identified. The non-carbonhydrate scaffold reported in this study suggests a new direction for development of bacterial transglycosylase inhibitors.
Assuntos
Antibacterianos/química , Glicosiltransferases/efeitos dos fármacos , Ensaios de Triagem em Larga Escala , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Antibacterianos/farmacologia , Concentração Inibidora 50 , Staphylococcus aureus Resistente à Meticilina/enzimologia , Testes de Sensibilidade Microbiana , Bibliotecas de Moléculas Pequenas , Infecções Estafilocócicas/tratamento farmacológico , Relação Estrutura-AtividadeRESUMO
The preparation of a novel fluorescent lipid II-based substrate for transglycosylases (TGases) is described. This substrate has characteristic structural features including a shorter lipid chain, a fluorophore tag at the end of the lipid chain rather than on the peptide chain, and no labeling with a radioactive atom. This fluorescent substrate is readily utilized in TGase activity assays to characterize TGases and also to evaluate the activities of TGase inhibitors.
Assuntos
Fluorescência , Glicosiltransferases/metabolismo , Fosfatos de Poli-Isoprenil/síntese química , Fosfatos de Poli-Isoprenil/metabolismo , Uridina Difosfato Ácido N-Acetilmurâmico/análogos & derivados , Bambermicinas/farmacologia , Inibidores Enzimáticos/farmacologia , Glicosiltransferases/antagonistas & inibidores , Estrutura Molecular , Fosfatos de Poli-Isoprenil/química , Relação Estrutura-Atividade , Especificidade por Substrato , Uridina Difosfato Ácido N-Acetilmurâmico/química , Uridina Difosfato Ácido N-Acetilmurâmico/metabolismoRESUMO
A practical and efficient solution-phase parallel synthesis of spirooxazolinoisoxazolines has been developed. Starting from methyl serine ester, the key steps are (1) acylation with concomitant beta-elimination to form an alpha,beta-unsaturated ester, (2) 1,3-dipolar cycloaddition with an oxime to form an isoxazoline ring, (3) reduction with NaBH(4), and (4) mesylation and in situ cyclization to form an oxazoline ring. All reaction steps and workup procedures were modified to allow the use of automated equipment including a synthesizer, a multifunctional liquid handler, and a vacuum centrifuge. Using this equipment, we synthesized a 100-membered library of spirooxazolinoisoxazoline in high yield, high purity, and excellent regioselectivity.
Assuntos
Técnicas de Química Combinatória/métodos , Oxazóis/química , Compostos de Espiro/química , Técnicas de Química Combinatória/economia , Oxazóis/síntese química , Soluções/química , Compostos de Espiro/síntese químicaRESUMO
Solid-phase organic synthesis of polyprenols with a traceless sulfone linker is described. The polymer-bound benezenesulfinate is first linked with the "tail" building blocks of isoprenyl chlorides via S-alkylation. With use of dimsyl anion as an appropriate base, the polymer-bound alpha-sulfonyl carbanion is generated and coupled with other "body" building blocks in an efficient manner. After repeated processes and a global palladium-catalyzed desulfonation with LiEt 3BH as the reducing agent, the desired polyprenols with various chain lengths and geometrical configurations are obtained in 32-59% overall yields. The solid-phase synthesis offers the advantage in facile isolation of polyprenols without tedious operation or time-consuming purification.
Assuntos
Pentanóis/síntese química , Polímeros/síntese química , Sulfonas/química , Hemiterpenos , Estrutura Molecular , Pentanóis/química , Polímeros/química , EstereoisomerismoRESUMO
The crystal structures of the dimer form of copper(II) tetraphenylporphyrin N-oxide, [Cu(tpp-N-O)]2 (3-dimer), and zinc(II) tetraphenylporphyrin N-oxide, [Zn(tpp-N-O)]2 (4-dimer), were established. The geometry at the copper ion in 3-dimer is essentially square-pyramidal with one oxygen bridge [O(1A)] occupying the apical site, giving a much larger Cu-O bond distance compared to those at the basal plane. The respective Cu...Cu distance and Cu-O-Cu angle in the core of 3-dimer are 3.987(4) A and 148.1(3) degrees. The Zn(1) atom in 4-dimer has a distorted square-pyramidal [4 + 1] coordination geometry that gives a tau-value of 0.19. The respective Zn...Zn distance and Zn-O-Zn angle in the dimeric unit of 4-dimer are 4.025(3) A and 148.1(2) degrees. The 3-dimer displays axial X-band electron paramagnetic resonance spectral features (Es = 0) in the powder state at 4 K, giving g parallel = 2.51 (A(parallel,s) = (9.6 +/- 0.2) x 10-3 cm(-1)) and g(perpendicular) = 2.11 and in the same powder state at 293 K giving Ds = 0.0731 cm(-1) (as derived from DeltaMs = 1 lines) or 0.0743 cm(-1) (as derived from the DeltaMs = 2 lines). In addition, 3-dimer displays a DeltaMs = 2 transition at g = 4.17 indicating the presence of spin-exchange coupling. The anisotropic exchange interaction (Ds(ex)= 0.132 cm(-1)) gives the main contribution to Ds in 3-dimer. The theoretical fit of the susceptibility and effective magnetic moment data of 3-dimer in the temperature range of 5-300 K gives 2J = 68 cm(-1), g = 2.01, p = 0.06, and a temperature-independent paramagnetism of 10(-6) cm3 mol(-1). This magnetic susceptibility data indicates that the copper(II) ions in 3-dimer are coupled in a ferromagnetic manner with the ground-spin triplet stabilized by 68 cm(-1) with regard to the singlet.