Darobactine mit Überlegener Antibiotischer Aktivität Generiert durch Strukturgeleitetes Kryo‐EM‐Biosynthese‐Design**

Seyfert, Carsten E.; Porten, Christoph; Yuan, Biao; Panter, Fabian; Bader, Chantal D.; Coetzee, Janetta; Deschner, Felix; Tehrani, Kamaleddin Haj Mohammad Ebrahim; Higgins, Paul G.; Seifert, Harald; Marlovits, Thomas C.; Herrmann, Jennifer; Müller, Rolf

doi:10.1002/ange.202214094

Cited by 2 publications

(6 citation statements)

References 41 publications

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“…Even though it was not detected in the HTS performed here, it shows high structural similarity and was reported from rationale-based DAR engineering approaches as a potent DAR derivative. (16,17,24) Table 1. Minimum inhibitory concentration of selected darobactin variants.…”

Section: Resultsmentioning

confidence: 99%

“…The minimum inhibitory concentration of the purified compounds were determined as previously described. (16,24) Briefly, overnight cultures of the test strains were cultivated at 37°C and 180rpm in Mueller Hinton II broth. For the BamA inhibitor sensitive E. coli MG1655 bamB::aac(3)-IV, we added 50 µg mL -1 apramycin and kanamycin to the culture medium.…”

Section: Mic Testingmentioning

confidence: 99%

“…(14,15) To engineer functional heterologous expression systems for new DAR-like BamA inhibitors only the precursor peptide and the modifying RaS enzyme are essential. (13,(16)(17)(18) The small core BGC (darAE) make the bicyclic heptapeptidic BamA inhibitors a prime target for mutasynthetic engineering. Certainly, not all theoretically possible 1.28 billion precursor heptamers will result in an active bicyclic molecule.…”

Section: Introductionmentioning

confidence: 99%

“…16,18) Additionally, a genome mining-based search for related RiPP structures that have the potential to inhibit the outer membrane protein BamA was sparked (12,16,42,43). Of these, the bicyclic heptameric DAR-like structures still represent the most promising class for antibiotic development.…”

mentioning

confidence: 99%

“…Hypothesis-driven bioengineering of the core peptide sequence helped formulate first SAR conclusions for BamA interacting with bicyclic heptapetides. The side chains of AAs, which are not directly involved in the DarE mediated macrocyclization, are promising moieties for optimization efforts (16)(17)(18). Sophisticated in silico design and selective in vitro bioactivity evaluation comprehensively summarized the impact of R2, R4 and R6 modifications as well as interacting effects among them on BamA bonding (45).…”

mentioning

confidence: 99%

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Nanoliter-scale selection of optimized bioengineered peptide antibiotics that rescue mice with bacterial lung infection

Böhringer,

Wuisan,

Marner

et al. 2024

Preprint

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Increasing numbers of multi-drug resistant pathogens call for new chemical scaffolds, addressing novel targets, that can serve as lead structures for the development of life-saving drugs. For antibiotics, natural product-inspired molecules represent a most promising resource. Natural products evolved to high chemical complexity and occupy a chemical space different than synthetic libraries. However, clinical translation of promising natural products is often impeded by their relative inaccessibility to medicinal chemistry optimization,e.g.iterative synthesis of large series of derivatives. Here, this limitation is addressed with a randomized library of bicyclic heptapeptides based on the natural product darobactin that hits the clinically not addressed target BamA. Variants of the ribosomally synthesized and post-translationally modified peptides were generated using heterologous mutasynthesis. A parallelized screening assay is adapted in nanoliter-scale beads to test the darobactin derivatives against our sensor strain. Loss of fluorescence sorting prioritized 563 events out of the analyzed ∼500k beads. Re-testing confirmed 48 hit events, of which 40 proved to produce distinct darobactin-type molecules. Most promising structures were isolated and the growth inhibitory effects against Gram-negative pathogens validated. One of our current frontrunner compounds (i.e., darobactin B) was reinforced by the randomized screen. While microbiological investigations of the new derivatives is ongoing, darobactin B was profiled in later tier assays and compared to another promising, rationally-designed analog (i.e., darobactin B9, “D22”). Early ADMET profiling and efficacy tests in a mouse pneumonia model were performed. Darobactin B reduced bacterial load ofPseudomonas aeruginosaandKlebsiella pneumoniaeby intraperitoneal, as well as intratracheal administration. Our study showcases the potential of mutasynthetic libraries for high-throughput screening and identification of functional peptides for drug lead discovery.

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Section: Resultsmentioning

confidence: 99%

Section: Mic Testingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Nanoliter-scale selection of optimized bioengineered peptide antibiotics that rescue mice with bacterial lung infection

Böhringer,

Wuisan,

Marner

et al. 2024

Preprint

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Fighting antibiotic resistance—strategies and (pre)clinical developments to find new antibacterials

et al. 2022

Self Cite

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Antibacterial resistance is one of the greatest threats to human health. The development of new therapeutics against bacterial pathogens has slowed drastically since the approvals of the first antibiotics in the early and mid-20 th century. Most of the currently investigated drug leads are modifications of approved antibacterials, many of which are derived from natural products. In this review, we highlight the challenges, advancements and current standing of the clinical and preclinical antibacterial research pipeline. Additionally, we present novel strategies for rejuvenating the discovery process and advocate for renewed and enthusiastic investment in the antibacterial discovery pipeline.

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Darobactine mit Überlegener Antibiotischer Aktivität Generiert durch Strukturgeleitetes Kryo‐EM‐Biosynthese‐Design**

Cited by 2 publications

References 41 publications

Nanoliter-scale selection of optimized bioengineered peptide antibiotics that rescue mice with bacterial lung infection

Nanoliter-scale selection of optimized bioengineered peptide antibiotics that rescue mice with bacterial lung infection

Fighting antibiotic resistance—strategies and (pre)clinical developments to find new antibacterials

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