Antimicrobial peptides (AMPs) have been identified as a potentially new class of antibiotics to combat bacterial resistance to conventional drugs. The design of de novo AMPs with high therapeutic indexes, low cost of synthesis, high resistance to proteases and high bioavailability remains a challenge. Such design requires computational modeling of antimicrobial properties. Currently, most computational methods cannot accurately calculate antimicrobial potency against particular strains of bacterial pathogens. We developed a tool for AMP prediction (Special Prediction (SP) tool) and made it available on our Web site (https://dbaasp.org/prediction). Based on this tool, a simple algorithm for the design of de novo AMPs (DSP) was created. We used DSP to design short peptides with high therapeutic indexes against gram-negative bacteria. The predicted peptides have been synthesized and tested in vitro against a panel of gram-negative bacteria, including drug resistant ones. Predicted activity against Escherichia coli ATCC 25922 was experimentally confirmed for 14 out of 15 peptides. Further improvements for designed peptides included the synthesis of D-enantiomers, which are traditionally used to increase resistance against proteases. One synthetic D-peptide (SP15D) possesses one of the lowest values of minimum inhibitory concentration (MIC) among all DBAASP database short peptides at the time of the submission of this article, while being highly stable against proteases and having a high therapeutic index. The mode of anti-bacterial action, assessed by fluorescence microscopy, shows that SP15D acts similarly to cell penetrating peptides. SP15D can be considered a promising candidate for the development of peptide antibiotics. We plan further exploratory studies with the SP tool, aiming at finding peptides which are active against other pathogenic organisms.

Antimicrobial peptides (AMPs) have emerged as promising candidates in combating antimicrobial resistance – a growing issue in healthcare. However, to develop AMPs into effective therapeutics, a thorough analysis and extensive investigations are essential. In this study, we employed an in silico approach to design cationic AMPs de novo, followed by their experimental testing. The antibacterial potential of de novo designed cationic AMPs, along with their synergistic properties in combination with conventional antibiotics was examined. Furthermore, the effects of bacterial inoculum density and metabolic state on the antibacterial activity of AMPs were evaluated. Finally, the impact of several potent AMPs on E. coli cell envelope and genomic DNA integrity was determined. Collectively, this comprehensive analysis provides insights into the unique characteristics of cationic AMPs.

Antimicrobial peptides (AMPs) have been identified as a potentially new class of antibiotics. There is a lot of computational methods of AMP prediction. Although most of them can predict antimicrobial potency against any microbe (microbe is not identified) with rather high accuracy, prediction quality of these tools against particular bacterial strains is low (Bioinformatics, 2018, in press; Journal of Chemical Information and Modeling 58, 1141-1151). Special prediction is a tool for the prediction of antimicrobial potency of peptides against particular target species with high accuracy. This tool is included into the Database of Antimicrobial Activity and Structure of Peptides (DBAASP, https://dbaasp.org, Nucleic acids research 44 (D1), D1104-D1112). In this presentation we describe this tool and predictive models for some Gram+ bacterial strains (Staphylococcus aureus ATCC 25923 and Bacillus subtilis) and a model for the prediction of hemolytic activity. Predictive model for Gram- Escherichia coli ATCC 25922 was presented earlier (Journal of Chemical Information and Modeling 58, 1141-1151, In 2nd Int. Electron. Conf. Med. Chem. 01-30 November 2016; Sciforum, 2016, A031). Special prediction tool can be used for the design of peptides being active against particular strain. To demonstrate the capability of the tool, peptides predicted as active against E. coli ATCC 25922 and S. aureus ATCC 25923 have been synthesized, and tested in vitro. The results have shown the justification of using special prediction tool for the design of new AMPs.

Emerging bacterial resistance to the existing antibiotics makes the development of new types of antibiotics an increasingly important challenge. Antimicrobial peptides (AMPs) can be considered as novel and efficient type of antibiotics that are hard to acquire resistance against. We have developed an algorithm to design peptides that are active against certain species. The prediction is based on clusterization of peptides with known biological activities by physicochemical properties. The Database of Antimicrobial Activity and Structure of Peptides (DBAASP, https://dbaasp.org) now includes Special Prediction (SP) tool, which allows to apply this algorithm to any amino acid sequence to predict whether this peptide is active against particular microbes. To verify the efficiency of the algorithm, we designed several variants of active peptides and tested them in vitro against two strains Escherichia coli ATCC 25922 and Staphylococcus aureus 25923. Prediction precision for the designed peptides against Escherichia coli ATCC was 95% and against Staphylococcus aureus was 68%. To improve prediction precision against Staphylococcus aureus we applied the linear regression analysis based on binary classification. This approach allows us to improve the prediction precision of the peptides designed for Staphylococcus aureus 25923 up to 92%.