RESUMO
Antiviral drug resistance in influenza infections has been a major threat to public health. To develop a broad-spectrum inhibitor of influenza to combat the problem of drug resistance, we previously identified the highly conserved E339...R416 salt bridge of the nucleoprotein trimer as a target and compound 1 as an inhibitor disrupting the salt bridge with an EC50 = 2.7 µM against influenza A (A/WSN/1933). We have further modified this compound via a structure-based approach and performed antiviral activity screening to identify compounds 29 and 30 with EC50 values of 110 and 120 nM, respectively, and without measurable host cell cytotoxicity. Compared to the clinically used neuraminidase inhibitors, these two compounds showed better activity profiles against drug-resistant influenza A strains, as well as influenza B, and improved survival of influenza-infected mice.
Assuntos
Compostos de Anilina/farmacologia , Antivirais/farmacologia , Vírus da Influenza A/química , Multimerização Proteica/efeitos dos fármacos , Proteínas de Ligação a RNA/metabolismo , Tiazóis/farmacologia , Proteínas do Core Viral/metabolismo , Compostos de Anilina/síntese química , Compostos de Anilina/metabolismo , Animais , Antivirais/síntese química , Antivirais/metabolismo , Sítios de Ligação/efeitos dos fármacos , Feminino , Camundongos Endogâmicos BALB C , Simulação de Acoplamento Molecular , Estrutura Molecular , Proteínas do Nucleocapsídeo , Ligação Proteica , Eletricidade Estática , Relação Estrutura-Atividade , Tiazóis/síntese química , Tiazóis/metabolismoRESUMO
Fluorinated glycosides are known to resist the glycosidase-catalyzed glycosidic bond cleavage; however, the synthesis of such glycans, especially 3-fluoro-sialic acid (3F-Neu5Ac) containing sialosides, has been a major challenge. Though the enzymatic synthesis of α-2,3-linked 3F-sialosides was reported, until recently there has not been any effective method available for the synthesis of 3F-sialosides in the α-2,6-linkage. In order to understand the biological effect of such modification, we report here a chemical synthesis of 3Fax-Neu5Ac-α2,6-Gal as a building block for the assembly of 3Fax-Neu5Ac-containing sialosides and a representative homogeneous antibody glycoform. Our results showed that the sialosides are stable under sialidase catalysis and the rituximab glycoform with a sialylated complex-type biantennary glycan terminated with 3Fax-Neu5Ac in the α-2,6-linkage (α2,6-F-SCT) has a similar binding avidity as its parent glycoform. These findings open up new opportunities for the development of therapeutic glycoproteins with improved pharmacokinetic parameters.
Assuntos
Anticorpos/química , Neuraminidase/química , Oligossacarídeos/síntese química , Ácidos Siálicos/química , Anticorpos/metabolismo , Biocatálise , Configuração de Carboidratos , Neuraminidase/metabolismo , Oligossacarídeos/química , Oligossacarídeos/metabolismo , Ácidos Siálicos/metabolismo , Especificidade por SubstratoRESUMO
The translation of biological glycosylation in humans to the clinical applications involves systematic studies using homogeneous samples of oligosaccharides and glycoconjugates, which could be accessed by chemical, enzymatic or other biological methods. However, the structural complexity and wide-range variations of glycans and their conjugates represent a major challenge in the synthesis of this class of biomolecules. To help navigate within many methods of oligosaccharide synthesis, this Perspective offers a critical assessment of the most promising synthetic strategies with an eye on the therapeutically relevant targets.
Assuntos
Oligossacarídeos/síntese química , Glicosilação , Humanos , Estrutura Molecular , Oligossacarídeos/químicaRESUMO
Described here is the asymmetric synthesis of iminosugar 2b, a Lipid II analog, designed to mimic the transition state of transglycosylation catalyzed by the bacterial transglycosylase. The high density of functional groups, together with a rich stereochemistry, represents an extraordinary challenge for chemical synthesis. The key 2,6-anti- stereochemistry of the iminosugar ring was established through an iridium-catalyzed asymmetric allylic amination. The developed synthetic route is suitable for the synthesis of focused libraries to enable the structure-activity relationship study and late-stage modification of iminosugar scaffold with variable lipid, peptide and sugar substituents. Compound 2b showed 70% inhibition of transglycosylase from Acinetobacter baumannii, providing a basis for further improvement.
Assuntos
Acinetobacter baumannii/efeitos dos fármacos , Antibacterianos/farmacologia , Inibidores Enzimáticos/farmacologia , Glicosiltransferases/antagonistas & inibidores , Uridina Difosfato Ácido N-Acetilmurâmico/análogos & derivados , Acinetobacter baumannii/enzimologia , Antibacterianos/síntese química , Antibacterianos/química , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/química , Glicosiltransferases/metabolismo , Testes de Sensibilidade Microbiana , Modelos Moleculares , Conformação Molecular , Estereoisomerismo , Relação Estrutura-Atividade , Uridina Difosfato Ácido N-Acetilmurâmico/síntese química , Uridina Difosfato Ácido N-Acetilmurâmico/química , Uridina Difosfato Ácido N-Acetilmurâmico/farmacologiaRESUMO
Glycosylation of lipids and proteins is not encoded by genes directly and depends on many factors including the origin of cell-lines, differential expression of carbohydrate enzymes and availability of substrates, as well as environmental conditions. Individual cells from different tissues produce each glycoprotein as heterogeneous mixtures of glycoforms with distinct biological activities in response to different conditions and disease states. As the result, the study of glycosylation could not rely purely on biochemical methods; instead it requires a multidisciplinary approach utilizing a variety of methods including genetic manipulation and glycosylation pathway engineering, structural and functional proteomic analysis, chemical and enzymatic synthesis, development of glycosylation probes and glycan microarrays. This review highlights recent progress and demonstrates how the availability of structure-defined oligosaccharides enables development of new and improved therapies, such as therapeutic homogeneous antibodies and carbohydrate-based vaccines against cancer.
Assuntos
Pesquisa Biomédica , Glicosilação , Polissacarídeos , Sequência de Carboidratos , Humanos , Análise em Microsséries , Modelos Moleculares , Técnicas de Sonda Molecular , VacinasRESUMO
Glycoscience research has been significantly impeded by the complex compositions of the glycans present in biological molecules and the lack of convenient tools suitable for studying the glycosylation process and its function. Polysaccharides and glycoconjugates are not encoded directly by genes; instead, their biosynthesis relies on the differential expression of carbohydrate enzymes, resulting in heterogeneous mixtures of glycoforms, each with a distinct physiological activity. Access to well-defined structures is required for functional study, and this has been provided by chemical and enzymatic synthesis and by the engineering of glycosylation pathways. This review covers general methods for preparing glycans commonly found in mammalian systems and applying them to the synthesis of therapeutically significant glycoconjugates (glycosaminoglycans, glycoproteins, glycolipids, glycosylphosphatidylinositol-anchored proteins) and the development of carbohydrate-based vaccines.
Assuntos
Glicoconjugados/síntese química , Glicoproteínas/síntese química , Glicosaminoglicanos/síntese química , Infecções por Haemophilus/prevenção & controle , Vacinas Anti-Haemophilus/administração & dosagem , Polissacarídeos/síntese química , Sequência de Aminoácidos , Configuração de Carboidratos , Sequência de Carboidratos , Glicoconjugados/imunologia , Glicolipídeos/síntese química , Glicolipídeos/imunologia , Glicoproteínas/imunologia , Glicosaminoglicanos/imunologia , Glicosilação , Glicosilfosfatidilinositóis/síntese química , Glicosilfosfatidilinositóis/imunologia , Infecções por Haemophilus/imunologia , Infecções por Haemophilus/microbiologia , Vacinas Anti-Haemophilus/síntese química , Haemophilus influenzae tipo b/efeitos dos fármacos , Haemophilus influenzae tipo b/crescimento & desenvolvimento , Haemophilus influenzae tipo b/patogenicidade , Humanos , Polissacarídeos/imunologiaRESUMO
Aryl sulfonyl chlorides (e.g. Ts-Cl) are beloved of organic chemists as the most commonly used S(VI) electrophiles, and the parent sulfuryl chloride, O2 S(VI) Cl2 , has also been relied on to create sulfates and sulfamides. However, the desired halide substitution event is often defeated by destruction of the sulfur electrophile because the S(VI) Cl bond is exceedingly sensitive to reductive collapse yielding S(IV) species and Cl(-) . Fortunately, the use of sulfur(VI) fluorides (e.g., R-SO2 -F and SO2 F2 ) leaves only the substitution pathway open. As with most of click chemistry, many essential features of sulfur(VI) fluoride reactivity were discovered long ago in Germany.6a Surprisingly, this extraordinary work faded from view rather abruptly in the mid-20th century. Here we seek to revive it, along with John Hyatt's unnoticed 1979 full paper exposition on CH2 CH-SO2 -F, the most perfect Michael acceptor ever found.98 To this history we add several new observations, including that the otherwise very stable gas SO2 F2 has excellent reactivity under the right circumstances. We also show that proton or silicon centers can activate the exchange of SF bonds for SO bonds to make functional products, and that the sulfate connector is surprisingly stable toward hydrolysis. Applications of this controllable ligation chemistry to small molecules, polymers, and biomolecules are discussed.
Assuntos
Química Click/métodos , Fluoretos/química , Compostos de Enxofre/química , Marcadores de Afinidade , Descoberta de Drogas , OxirreduçãoRESUMO
Molecules that bind selectively to a given protein and then undergo a rapid chemoselective reaction to form a covalent conjugate have utility in drug development. Herein a library of 1,3,4-oxadiazoles substituted at the 2 position with an aryl sulfonyl fluoride and at the 5 position with a substituted aryl known to have high affinity for the inner thyroxine binding subsite of transthyretin (TTR) was conceived of by structure-based design principles and was chemically synthesized. When bound in the thyroxine binding site, most of the aryl sulfonyl fluorides react rapidly and chemoselectively with the pKa-perturbed K15 residue, kinetically stabilizing TTR and thus preventing amyloid fibril formation, known to cause polyneuropathy. Conjugation t50s range from 1 to 4 min, ~1400 times faster than the hydrolysis reaction outside the thyroxine binding site. X-ray crystallography confirms the anticipated binding orientation and sheds light on the sulfonyl fluoride activation leading to the sulfonamide linkage to TTR. A few of the aryl sulfonyl fluorides efficiently form conjugates with TTR in plasma. Eleven of the TTR covalent kinetic stabilizers synthesized exhibit fluorescence upon conjugation and therefore could have imaging applications as a consequence of the environment sensitive fluorescence of the chromophore.
Assuntos
Amiloide/química , Flúor/química , Pré-Albumina/química , Multimerização Proteica/efeitos dos fármacos , Sulfonas/química , Sulfonas/farmacologia , Corantes Fluorescentes/química , Corantes Fluorescentes/farmacologia , Humanos , Cinética , Modelos Moleculares , Estabilidade Proteica/efeitos dos fármacos , Estrutura Secundária de ProteínaRESUMO
The recent development of a cell culture model of hepatitis C virus (HCV) infection based on the JFH-1 molecular clone has enabled discovery of new antiviral agents. Using a cell-based colorimetric screening assay to interrogate a 1,200-compound chemical library for anti-HCV activity, we identified a family of 1,2-diamines derived from trans-stilbene oxide that prevent HCV infection at nontoxic, low micromolar concentrations in cell culture. Structure-activity relationship analysis of ~ 300 derivatives synthesized using click chemistry yielded compounds with greatly enhanced low nanomolar potency and a > 1,000:1 therapeutic ratio. Using surrogate models of HCV infection, we showed that the compounds selectively block the initiation of replication of incoming HCV RNA but have no impact on viral entry, primary translation, or ongoing HCV RNA replication, nor do they suppress persistent HCV infection. Selection of an escape variant revealed that NS5A is directly or indirectly targeted by this compound. In summary, we have identified a family of HCV inhibitors that target a critical step in the establishment of HCV infection in which NS5A translated de novo from an incoming genomic HCV RNA template is required to initiate the replication of this important human pathogen.
Assuntos
Antivirais/farmacologia , Diaminas/farmacologia , Hepacivirus/efeitos dos fármacos , Estilbenos/farmacologia , Replicação Viral/efeitos dos fármacos , Antivirais/química , Antivirais/isolamento & purificação , Antivirais/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Diaminas/química , Diaminas/isolamento & purificação , Diaminas/toxicidade , Avaliação Pré-Clínica de Medicamentos/métodos , Farmacorresistência Viral , Hepatócitos/efeitos dos fármacos , Hepatócitos/virologia , Humanos , Testes de Sensibilidade Microbiana , RNA Viral/metabolismo , Estilbenos/química , Estilbenos/isolamento & purificação , Estilbenos/toxicidade , Relação Estrutura-Atividade , Proteínas não Estruturais Virais/genética , Proteínas não Estruturais Virais/metabolismoRESUMO
A new method for the synthesis of dihydroimidazo[1,2-a][1,3,5]triazin-4(6H)-ones via copper(I)-catalyzed hydroamination was developed. In addition, for the first time, iodoalkynes were shown to participate in the copper(I)-catalyzed intramolecular hydroamination reaction with exclusive formation of E-isomers.
Assuntos
Alcinos/química , Compostos Aza/síntese química , Cobre/química , Hidrocarbonetos Iodados/química , Purinas/síntese química , Aminação , Compostos Aza/química , Catálise , Isomerismo , Estrutura Molecular , Purinas/químicaRESUMO
[reaction: see text] The synthetic utility of alpha-halohydrazones is an underexplored area due to the lack of chemo- and regioselective routes towards these molecules. Herein, we describe a general method for alpha-bromohydrazone synthesis via the rearrangement of alpha-bromoaziridines, which can be readily prepared for the first time from the corresponding alkenyl bromides. The rearrangement of alpha-bromoaziridines into alpha-bromohydrazones proceeds with high yields and with high selectivities.
RESUMO
A variety of olefins were found to react with sulfinyl amides in the presence of POCl(3) to give beta-chlorosulfides and beta-hydroxysulfides in good yields. In the absence of nucleophiles, p-tolylsulfinyl amides were found to react with olefins with the formation of allylsulfoxides.