Ribosome-Templated Azide–Alkyne Cycloadditions: Synthesis of Potent Macrolide Antibiotics by In Situ Click Chemistry

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• 作者：Ian Glassford ; Christiana N. Teijaro ; Samer S. Daher ; Amy Weil ; Meagan C. Small ; Shiv K. Redhu ; Dennis J. Colussi ; Marlene A. Jacobson ; Wayne E. Childers ; Bettina Buttaro ; Allen W. Nicholson ; Alexander D. MacKerell Jr. ; Barry S. Cooperman ; Rodrigo B. Andrade
• 刊名：Journal of the American Chemical Society
• 出版年：2016
• 出版时间：March 9, 2016
• 年：2016
• 卷：138
• 期：9
• 页码：3136-3144
• 全文大小：542K
• ISSN：1520-5126

文摘

Over half of all antibiotics target the bacterial ribosome—nature’s complex, 2.5 MDa nanomachine responsible for decoding mRNA and synthesizing proteins. Macrolide antibiotics, exemplified by erythromycin, bind the 50S subunit with nM affinity and inhibit protein synthesis by blocking the passage of nascent oligopeptides. Solithromycin (1), a third-generation semisynthetic macrolide discovered by combinatorial copper-catalyzed click chemistry, was synthesized in situ by incubating either E. coli 70S ribosomes or 50S subunits with macrolide-functionalized azide 2 and 3-ethynylaniline (3) precursors. The ribosome-templated in situ click method was expanded from a binary reaction (i.e., one azide and one alkyne) to a six-component reaction (i.e., azide 2 and five alkynes) and ultimately to a 16-component reaction (i.e., azide 2 and 15 alkynes). The extent of triazole formation correlated with ribosome affinity for the anti (1,4)-regioisomers as revealed by measured K_d values. Computational analysis using the site-identification by ligand competitive saturation (SILCS) approach indicated that the relative affinity of the ligands was associated with the alteration of macrolactone+desosamine-ribosome interactions caused by the different alkynes. Protein synthesis inhibition experiments confirmed the mechanism of action. Evaluation of the minimal inhibitory concentrations (MIC) quantified the potency of the in situ click products and demonstrated the efficacy of this method in the triaging and prioritization of potent antibiotics that target the bacterial ribosome. Cell viability assays in human fibroblasts confirmed 2 and four analogues with therapeutic indices for bactericidal activity over in vitro mammalian cytotoxicity as essentially identical to solithromycin (1).