Lipopeptide Antibiotic Research Articles

Simulations of octapeptin–outer membrane interactions reveal conformational flexibility is linked to antimicrobial potency

The octapeptins are lipopeptide antibiotics that are structurally similar to polymyxins yet retain activity against polymyxin-resistant Gram-negative pathogens, suggesting they might be used to treat recalcitrant infections. However, the basis of their unique activity is unclear because of the difficulty in generating high-resolution experimental data of the interaction of antimicrobial peptides with lipid membranes. To elucidate these structure-activity relationships, we employed all-atom molecular dynamics simulations with umbrella sampling to investigate the conformational and energetic landscape of octapeptins interacting with bacterial outer membrane (OM). Specifically, we examined the interaction of octapeptin C4 and FADDI-115, lacking a single hydroxyl group compared with octapeptin C4, with the lipid A-phosphoethanolamine modified OM of Acinetobacter baumannii Octapeptin C4 and FADDI-115 both penetrated into the OM hydrophobic center but experienced different conformational transitions from an unfolded to a folded state that was highly dependent on the structural flexibility of their respective N-terminal fatty acyl groups. The additional hydroxyl group present in the fatty acyl group of octapeptin C4 resulted in the molecule becoming trapped in a semifolded state, leading to a higher free energy barrier for OM penetration. The free energy barrier for the translocation through the OM hydrophobic layer was ∼72 kcal/mol for octapeptin C4 and 62 kcal/mol for FADDI-115. Our results help to explain the lower antimicrobial activity previously observed for octapeptin C4 compared with FADDI-115 and more broadly improve our understanding of the structure-function relationships of octapeptins. These findings may facilitate the discovery of next-generation octapeptins against polymyxin-resistant Gram-negative 'superbugs.'

Pleiotropic regulation of daptomycin synthesis by DptR1, a LuxR family transcriptional regulator.

Daptomycin, produced by Streptomyces roseosporus is a novel cyclic lipopeptide antibiotic for treatment of Gram-positive bacteria caused infections. While, the regulatory mechanism of daptomycin synthesis has not been fully understood. Here we reported that DptR1, a LuxR family transcriptional regulator, played a pleiotropic regulatory role on daptomycin synthesis, for the first time. Deletion or over-expressing of dptR1 decreases the daptomycin's production, increases the transcriptional levels of the core dpt genes of day 3 and decreases the transcriptional levels of the core dpt genes of day 4, sharply, which indicates the transcriptional regulation of DptR1 on daptomycin synthesis is complex and time-ordered. The transcriptional levels of dptR2 increase in dptR1 deletion mutant (DR1), but decrease in dptR1 over-expression mutant (OR1), dramatically, compared to the starting strain of Streptomyces roseosporus N3 (WT), on the 3rd day, which indicates that DptR1 represses the transcription of dptR2. While, the transcriptional levels of dptR3 both in DR1 and OR1 decrease obviously, compared to WT, on the 3rd and 4th day. Comparative analysis of promoters' activities, using xylE gene as the reporter, showed that DptR1 activated the transcription of its own gene of dptR1 and represses the transcription of the dptR3 by affecting the promoter activities. While DptR1 may affect the expression of dptR2 indirectly, not by affecting the promoter activity of dptR2. DptR1, a LuxR family transcriptional regulator, played a pleiotropic regulation role on daptomycin synthesis.

Implications of Glycopeptide and Lipopeptide Antibiotics on Asthma Exacerbations in a Patient With Hyper-IgE Syndrome and Chronic Severe Asthma: A Case Report

Objective: To review and consider the immunomodulatory ramifications of glycopeptide and lipopeptide antibiotics in a patient with severe asthma, hyper-immunoglobulin E (IgE) syndrome, and osteonecrosis of the jaw. Case Summary: A 36-year-old male patient with a pertinent past medical history of hyper-IgE syndrome, severe asthma, and osteonecrosis of the jaw attributed to steroid use initially presented to the emergency department, where imaging of the jaw suggested an infectious process. Following months of vancomycin therapy as an outpatient, the patient was switched to daptomycin to facilitate once-daily infusions. Following this change, the patient experienced significantly less asthma exacerbations and fewer admissions to the hospital for asthma-related issues. Discussion: Though daptomycin is associated with eosinophilia, and this patient’s eosinophil counts did increase while on the drug, an inverse relationship developed concerning the number of hospital admissions for asthma exacerbations. A review of the literature indicates that select glycopeptides, such as vancomycin, have pro-inflammatory effects, while other related drugs, such as the lipopeptide daptomycin, may not result in clinical manifestations of an inflammatory process. Tumor necrosis factor-α, interleukin-1β, and interleukin-6 have been implicated in this process. This patient may have been particularly susceptible to the effects of the glycopeptides due to his diagnosis of hyper-IgE syndrome, which has been independently associated with reactive airway diseases. Conclusion: Cytokine migration and activation by select glycopeptide and lipopeptide antibiotics warrant consideration from clinicians when determining the most appropriate treatment regimens. Further research is required to elucidate whether the pro-inflammatory properties of vancomycin warrant a change in first-line therapy to daptomycin in select cases.

Novel Cyclic Lipopeptide Antibiotics: Effects of Acyl Chain Length and Position.

Multidrug-resistant bacteria are a global health problem. One of the last-resort antibiotics against Gram-negative bacteria is the cyclic lipopeptide colistin, displaying a flexible linker with a fatty acid moiety. The aim of the present project was to investigate the effect on antimicrobial activity of introducing fatty acid moieties of different lengths and in different positions in a cyclic peptide, S3(B), containing a flexible linker. The lipidated analogues of S3(B) were synthesized by 9-fluorenylmethoxycarbonyl (Fmoc) solid-phase peptide synthesis. Following assembly of the linear peptide by Fmoc solid-phase peptide synthesis, on-resin head-to-tail cyclization and fatty acid acylation were performed. The antimicrobial activity was determined against the ESKAPE pathogens, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Escherichia coli. Furthermore, hemolytic activity was determined against human erythrocytes. A total of 18 cyclic lipopeptides were synthesized and characterized. It was found that introduction of fatty acids in positions next to the flexible linker was more strongly linked to antimicrobial activity. The fatty acid length altered the overall hydrophobicity, which was the driving force for both high antimicrobial and hemolytic activity. Peptides became highly hemolytic when carbon-chain length exceeded 10 (i.e., C10), overlapping with the optimum for antimicrobial activity (i.e., C8–C12). The most promising candidate (C8)5 showed antimicrobial activity corresponding to that of S3(B), but with an improved hemolytic profile. Finally, (C8)5 was further investigated in a time-kill experiment.

Pleiotropic regulation of daptomycin synthesis by DptR1, a LuxR family transcriptional regulator.

Daptomycin, produced by Streptomyces roseosporus is a novel cyclic lipopeptide antibiotic for treatment of Gram-positive bacteria caused infections. While, the regulatory mechanism of daptomycin synthesis has not been fully understood. Here we reported that DptR1, a LuxR family transcriptional regulator, played a pleiotropic regulatory role on daptomycin synthesis, for the first time. Deletion or over-expressing of dptR1 decreases the daptomycin's production, increases the transcriptional levels of the core dpt genes of day 3 and decreases the transcriptional levels of the core dpt genes of day 4, sharply, which indicates the transcriptional regulation of DptR1 on daptomycin synthesis is complex and time-ordered. The transcriptional levels of dptR2 increase in dptR1 deletion mutant (DR1), but decrease in dptR1 over-expression mutant (OR1), dramatically, compared to the starting strain of Streptomyces roseosporus N3 (WT), on the 3rd day, which indicates that DptR1 represses the transcription of dptR2. While, the transcriptional levels of dptR3 both in DR1 and OR1 decrease obviously, compared to WT, on the 3rd and 4th day. Comparative analysis of promoters' activities, using xylE gene as the reporter, shows that DptR1 activated the transcription of its own gene of dptR1 and represses the transcription of the dptR3 by affecting the promoter activities. While DptR1 may affect the expression of dptR2 indirectly, not by affecting the promoter activity of dptR2. DptR1, a LuxR family transcriptional regulator, played a pleiotropic regulation role on daptomycin synthesis.

Screening of Bacillus strains in biocontrol of pathogen Dickeya dadantii causing stem and root rot disease of sweet potato

ABSTRACT Outbreaks of stem and root rot disease of sweet potato caused by Dickeya dadantii recently occurred in major sweet potato planting areas in China. This study aimed to screen effective Bacillus strains in biocontrol of the bacterial soft rot pathogen. We screened plant-associated Bacillus strains capable of inhibiting D. dadantii using an in vitro overlay culture assay and an in vivo tuber slice assay. All Bacillus strains inhibited D. dadantii growth and maceration of sweet potato tubers. Two strains B. amyloliquefaciens A3 and B. velezensis A2 showed significantly higher rates of in vitro inhibition (55% and 50%) on D. dadantii growth and in vivo reduction (82% and 77%) of the maceration of sweet potato tuber slices by D. dadantii than other Bacillus strains did. They showed broad-spectrum inhibition on growth of soft rot Dickeya and Pectobacterium strains. They protected sweet potato seed tubers from infection by D. dadantii and promoted seedling growth. Their culture supernatants inhibited D. dadantii growth, swimming and biofilms and damaged D. dadantii cells. B. amyloliquefaciens A3 is more effective than B. velezensis A2 in producing surfactins, well-known lipopeptide antibiotics and biosurfactants, and inhibiting D. dadantii. Together, we developed an effective method to screen effective Bacillus strains to control the soft rot pathogen D. dadantii and screened out a strong surfactin producer B. amyloliquefaciens A3, which can be used as a reference strain for further screening of more effective Bacillus strains and to develop an agent to control the stem and root rot disease of sweet potato.

Structure-Activity Relationships of Daptomycin Lipopeptides.

Daptomycin is a calcium-dependent cyclic lipodepsipeptide derived from the soil saprotroph Streptomyces roseosporus, and its antibiotic properties make it a key agent for treatment of drug-resistant Gram-positive infections. It is most commonly used clinically for the treatment of Gram-positive skin and skin structure infections (SSSI), Staphylococcus aureus bacteremia, and right-sided endocarditis infections associated with S. aureus, including methicillin resistant S. aureus (MRSA). It has also been used "off-label" for Enterococcal infections. There has been a tremendous amount of research investigating its mode of action, resistance mechanisms, and biosynthesis of this clinically important antimicrobial agent. Although we cover the latter aspects in detail, the primary focus of this review is to provide the most comprehensive and up-to-date reference for the medicinal chemist on the structure-activity-toxicity of this important class of lipopeptide antibiotics.

Evaluation of Daptomycin-Induced Cellular Membrane Injury in Skeletal Muscle.

Daptomycin, a cyclic lipopeptide antibiotic, has bactericidal activity against Gram-positive organisms and is especially effective against methicillin-resistant Staphylococcus aureus. Although daptomycin causes unique adverse drug reactions such as elevation of creatine phosphokinase or rhabdomyolysis, the detailed mechanisms underlying these adverse drug reactions in skeletal muscle are unclear. This study aimed to elucidate whether daptomycin causes direct skeletal muscle cell toxicity and investigate the relationship between daptomycin exposure and musculoskeletal toxicity. First, we evaluated the relationship between daptomycin exposure and skeletal muscle toxicity. Of the 38 patients who received daptomycin intravenously, an elevation in creatine phosphokinase levels was observed in five. The median plasma trough concentration of daptomycin in patients with elevated creatine phosphokinase levels was significantly higher than that in patients whose creatine phosphokinase levels were within the normal range, suggesting that increased exposure to daptomycin is related to elevation in creatine phosphokinase levels. In an in vitro study using human rhabdomyosarcoma cells, daptomycin reduced cell viability and increased membrane damage. These effects were more marked under hypoxic conditions. A necroptotic pathway seemed to be involved because phosphorylated mixed lineage kinase domain-like protein expression was enhanced following daptomycin exposure, which was significantly enhanced under hypoxic conditions. These findings indicate that daptomycin elicits cytotoxic effects against skeletal muscle cells via the necroptotic pathway, and the extent of toxicity is enhanced under hypoxic conditions.

Characterization of the Interaction of Daptomycin With Site II on Human Serum Albumin

Daptomycin, a cyclic lipopeptide antibiotic, is clinically used for the treatment of infections caused by Gram-positive bacteria, including the methicillin-resistant Staphylococcus aureus and the vancomycin-resistant Enterococci. While daptomycin shows high plasma protein binding (90–93%), our knowledge of the binding process is not extensive. To address this issue in more detail, we characterized the binding of daptomycin to plasma proteins and the findings indicate that the association constant for the binding of daptomycin to human serum albumin (HSA) is much higher than that for α1-acid glycoprotein, another plasma protein. Daptomycin was also found to bind to a single site on HSA, which was identified as site II. The findings also suggest that the n-decanoyl moiety of daptomycin penetrates into the hydrophobic pocket of site II and that this acyl moiety interacts with Tyr411 at the entrance to site II. Due to this selective interaction with site II, daptomycin binding was significantly inhibited by drugs (ibuprofen or diazepam) and endogenous compounds (uremic toxins or fatty acids) which also strongly bind to site II. In diseased states, such an inhibition in the binding could result in the pharmacokinetics and therapeutic action of daptomycin being substantially altered.

Role of Biofilm Formation by Bacillus pumilus HR10 in Biocontrol against Pine Seedling Damping-Off Disease Caused by Rhizoctonia solani

The biocontrol process mediated by plant growth-promoting rhizobacteria (PGPR) relies on multiple mechanisms. Biofilm formation plays an important role in the ability of PGPR to control plant diseases. Bacillus pumilus HR10, one such PGPR, promotes the growth of Pinus thunbergii. This study showed that the wild-type strain B. pumilus HR10 produces a stable and mature biofilm in vitro. Biofilm-deficient mutants of B. pumilus HR10 with different phenotypes were screened by mutagenesis. The contents of extracellular polysaccharides (EPS) and proteins produced by the mutant strains were significantly reduced, and the biofilms of the mutants were weakened to varying degrees. The swarming abilities of the wild-type and mutant strains were positively correlated with biofilm formation. A colonization assay demonstrated that B. pumilus HR10 could colonize the roots of Pinus massoniana seedlings in a large population and persist, while biofilm-deficient mutants showed weak colonization ability. Furthermore, a biocontrol assay showed that biocontrol efficacy of the mutants was reduced to a certain degree. We determined the inhibitory activity of B. pumilus HR10 and its ability to induce systemic resistance against Rhizoctonia solani of plants. The synthesis of lipopeptide antibiotics is probably involved in biofilm formation by B. pumilus HR10. These observations not only provide a reference for further research about the coordinated action between biofilm formation and the multiple biocontrol mechanisms of B. pumilus HR10 but also improve the understanding of the regulatory pathway of biofilm formation by B. pumilus HR10.

Stalobacin: Discovery of Novel Lipopeptide Antibiotics with Potent Antibacterial Activity against Multidrug-Resistant Bacteria.

A novel lipopeptide antibiotic, stalobacin I (1), was discovered from a culture broth of an unidentified Gram-negative bacterium. Stalobacin I (1) had a unique chemical architecture composed of an upper and a lower half peptide sequence, which were linked via a hemiaminal methylene moiety. The sequence of 1 contained an unusual amino acid, carnosadine, 3,4-dihydroxyariginine, 3-hydroxyisoleucine, and 3-hydroxyaspartic acid, and a novel cyclopropyl fatty acid. The antibacterial activity of 1 against a broad range of drug-resistant Gram-positive bacteria was much stronger than those of "last resort" antibiotics such as vancomycin, linezolid, and telavancin (MIC 0.004-0.016 μg/mL). Furthermore, compound 1 induced a characteristic morphological change in Gram-positive and Gram-negative strains by inflating the bacterial cell body. The absolute configuration of a cyclopropyl amino acid, carnosadine, was determined by the synthetic study of its stereoisomers, which was an essential component for the strong activity of 1.

Design, Synthesis, and Bioactivity of Cyclic Lipopeptide Antibiotics with Varied Polarity, Hydrophobicity, and Positive Charge Distribution.

Twenty-three polymyxin analogs with variations at nine amino acid positions were synthesized and assessed for antimicrobial activity and renal cytotoxicity. Compounds M2, 14, S2, and 16 (MIC = 0.125-4 μg/mL) had similar or stronger activities against susceptible and drug-resistant strains of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii compared to polymyxin B (MIC = 1-2 μg/mL). Most synthesized compounds (50% cytotoxic concentration, CC50 ≥ 200 μg/mL) exhibited lower cytotoxicity than polymyxin B (CC50 = 99 ± 6 μg/mL). Polymyxin S2 showed high plasma stability in vitro and strong efficacy in a mouse systemic infection model (ED50 = 0.9 mg/kg) against NDM-1-producing Klebsiella pneumoniae, suggesting that it is a potential candidate for drug development. The activity and cytotoxicity results indicated that the amino acids at positions 2, 3, 6, and 7 might be replaced. Effects on activity and cytotoxicity linked to changes in the number of positively charged amino acids varied among different cyclopeptide skeletons, but the underlying mechanisms are unknown.

Daptomycin susceptibility in enterococci isolated from urinary samples in a tertiary care hospital

are gram positive cocci encountered in various human infections.The most striking feature of is their relative and absolute resistance to variety of antibiotics.Vancomycin resistant (VRE) were first described in late 1980 and the incidence of nosocomial infections by VRE being 11.9% in Asia Pacific and 35.5% in U.S. Daptomycin is a lipopeptide antibiotic with bactericidal activity . To study the daptomycin susceptibility in isolated in urinary samples. A total of 76 strains of were isolated during the study period.25 (33%) were identified as and 51 (67%) as Most of the Enterococcal isolates (82.8%) were resistant to Norfloxacin followed by 71% being resistant to tetracycline and 76% of isolates were sensitive to nitrofurantoin. Most of the isolates were multidrug resistant. Of the 76 isolated only 2(2.63%) were VRE and both the VRE strains isolated were . There was no resistance to linezolid including the VRE strains. All the isolates 76 (100%) were sensitive to daptomycin including the VRE strains. Our study mainly focused on susceptibility of enterococcal isolates in urinary samples to daptomycin which has bactericidal activity against gram positive organisms including multidrug resistant strains. This study shows growing concern for increasing antibiotic resistance in enterococcal isolates and treatment of these infections with daptomycin is invited as there are only few reports of daptomycin non susceptible (DNSE).

Characterization of multimeric daptomycin bound to lipid nanodiscs formed by calcium-tolerant styrene-maleic acid copolymer

Daptomycin is a lipopeptide antibiotic that is important in the treatment of infections with Gram-positive bacteria. In the presence of calcium, daptomycin binds to phosphatidylglycerol in the bacterial cytoplasmic membrane and then forms oligomers that mediate its bactericidal effect. The structure of these bactericidal oligomers has not been elucidated. We here explore the feasibility of structural studies on the oligomer by solution-state NMR. To this end, we use nanodiscs that contain DMPC and DMPG, stabilized with a styrene-maleic acid copolymer that has been modified to minimize calcium chelation. We show that these nanodiscs bind daptomycin and induce the formation of stable oligomers under physiologically relevant conditions. The findings suggest that this membrane model is suitable for structural and functional characterization of oligomeric daptomycin, and possibly of other calcium-dependent lipopeptide antibiotics. We show that these nanodiscs bind daptomycin and induce the formation of stable oligomers, under conditions that are suitable for biomolecular NMR. The findings suggest that this membrane model is suitable for structural elucidation of oligomeric daptomycin, and possibly of other calcium-dependent lipopeptide antibiotics.

An endophytic strain of the genus Bacillus isolated from the seeds of maize (Zea mays L.) has antagonistic activity against maize pathogenic strains

Plant endophytes are microbes that colonize plant internal tissues and are ubiquitously associated with plants. In this study, seven endophytic bacterial strains, 665L2, 725L2, 725R2, 92R2, 728R3, 728R4 and 2416T3, were isolated from seeds of five healthy maize varieties (Zea mays L.) and all identified as Bacillus velezensis by polyphasic taxonomy based on 16S rRNA and gyrA gene phylogenetic analysis. In addition, according to the genotyping results from random amplified polymorphic DNA (RAPD), 665L2, 725L2, 725R2 and 92R2 belonged to the same strain, while 728R3 and 2416T3 belonged to another strain. Pathogenic fungal strains 4, 5 and 6 were isolated from three diseased maize varieties (Zea mays L.), and they were identified as Talaromyces funiculosus, Penicillium oxalicum and Fusarium verticillioides, respectively, by polyphasic taxonomy based on morphological identification, ITS rDNA and bio-control gene phylogenetic analyses. Seven endophytic bacterial Bacillus velezensis strains had favourable antagonistic activity, and antagonistic testing was carried out against the three pathogenic strains, Talaromyces funiculosus 4, Penicillium oxalicum 5 and Fusarium verticillioides 6. Biological control lipopeptide antibiotic genes (bioA, bmyB, ituC, fenD, srfAA, srfAB, yngG and yndJ) were amplified using specific primers, and they were found in the seven endophytic bacterial Bacillus velezensis strains. This study provides a scientific basis for future research on the use of resistant endophytic bacterial resources to enhance crop production.

A new validated HPLC-UV method for therapeutic monitoring of daptomycin in comparison with reference mass spectrometry

Daptomycin, a cyclic lipopeptide antibiotic with a broad spectrum of activity against Gram-positive bacteria, is also active against multi-resistant bacterial strains, as well as methicillin-resistant S. aureus, vancomycin-resistant enterococci or penicillin-resistant S. pneumoniae. For these reasons it is a viable alternative for the treatment of persisting infections. However, the therapeutic drug monitoring of daptomycin is recommended because the known variability in drug disposition and the severe clinical conditions of patients. Therefore, we developed a simple and fast UV-HPLC method according to FDA guidelines to monitor plasma concentrations of the drug. Briefly, after a liquid-liquid extraction, plasma calibration samples, quality controls and patients’ samples were injected in a HPLC instrument and peaks of daptomycin and gentamicin (internal standard) were resolved by a C18 250 × 4.6 mm, 5 μm stationary phase and peaks were monitored at UV = 262 nm. Mobile phase (isocratic flow of 1 mL/min) consisted of acetonitrile-buffer (KH2PO4 20 mM pH = 3.2) 46:54, vol/vol. Under these conditions, IS and daptomycin peaked at 4.1 and 5.8 min after injection. Values of limits of detection and quantitation accounted for 1.65 and 5.00 (μg/ml), respectively. Values of method linearity (r2) in range 5−100 mg/L were 0.9975 and 0.9956 plasma samples and solvent standard, respectively. Inter- and intra-day variability coefficients were lower than 15 %. The comparison with a reference, commercially-available LC–MS/MS method on 122 patient plasma samples returned excellent correlation (r2 = 0.9474). In conclusion, the present method demonstrated to be reliable and suitable for daptomycin TDM in clinical routine.

More Than a Pore: A Current Perspective on the In Vivo Mode of Action of the Lipopeptide Antibiotic Daptomycin.

Daptomycin is a cyclic lipopeptide antibiotic, which was discovered in 1987 and entered the market in 2003. To date, it serves as last resort antibiotic to treat complicated skin infections, bacteremia, and right-sided endocarditis caused by Gram-positive pathogens, most prominently methicillin-resistant Staphylococcus aureus. Daptomycin was the last representative of a novel antibiotic class that was introduced to the clinic. It is also one of the few membrane-active compounds that can be applied systemically. While membrane-active antibiotics have long been limited to topical applications and were generally excluded from systemic drug development, they promise slower resistance development than many classical drugs that target single proteins. The success of daptomycin together with the emergence of more and more multi-resistant superbugs attracted renewed interest in this compound class. Studying daptomycin as a pioneering systemic membrane-active compound might help to pave the way for future membrane-targeting antibiotics. However, more than 30 years after its discovery, the exact mechanism of action of daptomycin is still debated. In particular, there is a prominent discrepancy between in vivo and in vitro studies. In this review, we discuss the current knowledge on the mechanism of daptomycin against Gram-positive bacteria and try to offer explanations for these conflicting observations.

Hypothesis: daptomycin permeabilizes membranes by forming self assembled nanotubes

Daptomycin is the only lipopeptide antibiotic that is widely used in clinical practice. It was discovered by Eli Lilly and then studied and commercialized by Cubist Pharmaceuticals in 2003. Although this antibiotic has been used for 17 years, the debate over its mechanism of action is ongoing. In this paper, we discuss the different hypotheses on the mode of action of this antibiotic with a primary focus on the bacterial membrane permeabilization as the main mechanism of action. By comparing the experimental data on the oligomerization of daptomycin in membranes with properties of self-assembling cyclic peptides, we conclude that the structure of daptomycin oligomer should resemble the structures of peptide nanotubes that serve as ion channels in membranes.

The contribution of achiral residues in the laspartomycin family of calcium-dependent lipopeptide antibiotics.

The growing threat of antibacterial resistance is a global concern. The so-called calcium-dependent lipopeptide antibiotics (CDAs) have emerged as a promising source of new antibiotic agents that are rich in structural and mechanistic diversity. Over forty unique CDAs have been identified to date and share a number of common features. Recent efforts in our group have provided new mechanistic and structural insights into the laspartomycin family of CDAs. We here describe investigations aimed at probing the role of the three glycine residues found in the laspartomycin peptide macrocycle. In doing so laspartomycin analogues containing the achiral 2-aminoisobutyric acid (AIB) as well as l- or d-alanine in place of glycine were prepared and their antibacterial activities evaluated.

Solid-Phase Synthesis of Octapeptin Lipopeptides.

Octapeptins are naturally derived cyclic lipopeptide antibiotics with activity against a range of Gram-negative pathogens, including highly resistant strains. Octapeptin C4, an exemplar of the class, was synthesized using a combination of Fmoc solid-phase peptide synthesis (SPPS) and solution-phase cyclization. Utilizing H-L-Leu-2-chlorotrityl resin, peptide couplings were performed using HCTU and collidine in DMF. The linear sequence was terminated by N-acylation with 3-(R)-hydroxydecanoic acid. The residue Dab-2 was orthogonally protected with 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)isovaleryl group (ivDde) to enable selective side-chain deprotection prior to resin cleavage. Resin cleavage was accomplished with hexafluoroisopropanol in DCM, followed by cyclization with diphenylphosphoryl azide (DPPA) and solid sodium bicarbonate in DMF.