Antimicrobial peptides (AMPs) are a primary defense against pathogens. Here, we examined the interaction of two BP100 analogs, R²R⁵-BP100 (where Arg substitutes Lys 2 and 5) and R²R⁵-BP100-A-NH-C₁₆ (where an Ala and a C₁₆ hydrocarbon chain are added to the R²R⁵-BP100 C-terminus), with membrane models. Large unilamellar vesicles (LUVs) and giant unilamellar vesicles (GUVs) were prepared with the major lipids in Gram-positive (GP) and Gram-negative (GN) bacteria, as well as red blood cells (RBCs). Fluorescence data, dynamic light scattering (DLS), and zeta potential measurements revealed that upon achieving electroneutrality through peptide binding, vesicle aggregation occurred. Circular dichroism (CD) spectra corroborated these observations, and upon vesicle binding, the peptides acquired α-helical conformation. The peptide concentration, producing a 50% release of carboxyfluorescein (C₅₀) from LUVs, was similar for GP-LUVs. With GN and RBC-LUVs, C₅₀ decreased in the following order: BP100 > R²R⁵-BP100 > R²R⁵BP100-A-NH-C₁₆. Optical microscopy of GP-, GN-, and RBC-GUVs revealed the rupture or bursting of the two former membranes, consistent with a carpet mechanism of action. Using GUVs, we confirmed RBC aggregation by BP100 and R²R⁵-BP100. We determined the minimal inhibitory concentrations (MICs) of peptides for a GN bacterium (Escherichia coli (E. coli)) and two GP bacteria (two strains of Staphylococcus aureus (S. aureus) and one strain of Bacillus subtilis (B. subtilis)). The MICs for S. aureus were strain-dependent. These results demonstrate that Lys/Arg replacement can improve the parent peptide’s antimicrobial activity while increasing hydrophobicity renders the peptide less effective and more hemolytic.

Background: 2-[¹⁸F]Fluoroethyltosylate ([¹⁸F]FEtOTs) is a well-known¹⁸F-fluoroalkylating agent widely used to synthesize radiotracers for positron emission tomography. The widespread use of [¹⁸F]FEtOTs is due in part to its low volatility when compared to other halide and sulfonate building blocks. In this work, the radioactive volatile side-products formed during the synthesis of [¹⁸F]FEtOTs were identified and characterized for the first time, and an optimization of the reaction conditions to minimize their formation was proposed.Results: In order to characterize the volatiles produced during [¹⁸F]FEtOTs synthesis, the reaction mixtures of both cold FEtOTs and [¹⁸F]FEtOTs were co-injected onto the HPLC system. The radioactive peaks corresponding to the volatile compounds were collected, analyzed through headspace gas chromatography mass spectrometry sampler (HS-GC–MS) and identified as vinyl fluoride ([¹⁹F]VF) and 2-fluoroethanol ([¹⁹F]FEOH). By using a rotatable central composite design with a two-level full factorial core of two factors (2²), it was determined that temperature and time are independent variables which affect the generation of [¹⁸F]VF and [¹⁸F]FEOH during the radiosynthesis of [¹⁸F]FEtOTs. In addition, in order to reduce the formation of the volatiles ([¹⁸F]VF and [¹⁸F]FEOH) and increase the yield of [¹⁸F]FEtOTs, it was demonstrated that the molar ratio of base to precursor must also be considered.Conclusion: [¹⁸F]VF and [¹⁸F]FEOH are volatile side-products formed during the radiosynthesis of [¹⁸F]FEtOTs, whose yields depend on the reaction time, temperature, and the molar ratio of base to precursor. Therefore, special care should be taken during the radiosynthesis and subsequent reactions using [¹⁸F]FEOTs in order to avoid environmental contamination and to improve the yield of the desired products.