Title Background To date, the wide spreading of the ever-increasing number of antibiotic-resistant bacterial strain is a serious healthcare problem,

and creates urgent needs for novel antibacterial compounds. Thus, the design and the synthesis of new antimicrobial compounds is a major challenge in medicinal chemistry research and, in this context, a role of primary importance is played by antimicrobial peptides and their synthetic derivatives. Aims Herein is presented the designed and synthesized new small antimicrobial peptidomimetic derivatives, using the dipeptide L-arginine-L-tryptophane benzyl ester, reported by Strm et al. [17] as model compound. Methods In order to obtain structure-activity relationships, have been designed two series of SAMPs comprising arginine analogues and a set of aromatic amino acids in their structure. The synthesized compounds were tested against Gram positive (Staphylococcus aureus ATCC 29213 and Staphylococcus epidermidis ATCC 35984) and Gram negative (Escherichia coli ATCC 8739 and Klebsiella pneumoniae 5F) bacterial strains and one yeast (Candida albicans ATCC 10231), all biofilm producers. Furthermore, for the most active compounds were estimated also the antibiofilm activity against Staphylococcal biofilm. The toxicity of the compounds was also estimated, determining their hemolytic activity against human red blood cells. Results The synthesized compounds 1-8 were reported in Figure 1. The antimicrobial activity of the peptidomimetics 1-8 and of the reference compound ArgTrpOBzl were determined against two reference Gram positive strains (S. aureus and S. epidermidis), two reference Gram negative strain (E. coli and K. pneumoniae) and a yeast (C. albicans). The minimum inhibitory concentration (MIC) values against Gram positive species was ranged from 300 to 7.5 g/ml. The compounds 5 and 6 showed the best antibacterial effect both against S. aureus (10 g/ml) and S. epidermidis (7.5 g/mL), while the compounds 1 and 2 are less active, showing MIC values >25 g/mL. The bactericidal effect (MBC) for these bacterial strains was ranged from 300 to 10 g/mL, with the same value as MIC recorded in the compounds 5, 1 and 3 against S. aureus, compound 7 against S. epidermidis, compounds 3, 7 and 8 against E. coli. The MIC values obtained against E. coli were higher than that obtained with Gram positive strains, ranging from 300 to 25 g/ml. In particular, compounds 2 and 5 displayed the best antibacterial activity respectively against K. pneumoniae (31.3 g/mL) and E. coli (25 g/mL). Both these compounds displayed the highest activity against C. albicans (50 and 62.5 g/mL, respectively). Each compound tested displayed no significant differences between MIC and MBC values. Considering the lower activity against E. coli respect to Gram positive species, the effect of peptides on established biofilms was evaluated only against S. aureus and S. epidermidis. Results showed BIC values ranging from four-fold to twenty-fold higher than the corresponding MIC values, while biofilm eradicating concentrations (BEC) were never under 800 g/mL for all peptides tested. Thus, all synthesized peptidomimetics 1-8, as well as the model compound ArgTrpOBzl, showed a fair inhibiting activity on established Gram-positive biofilms

(BIC < 200), but a lower capacity to eradicate the biofilms (BEC > 800). All analyzed compounds did not produce the human blood hemolysis at their MIC values. Figure 1

Conclusion In conclusion, novel antimicrobial peptidomimetics containing the 1-(4(aminomethyl)benzyl)guanidine or the 3,5-diguanidino benzoic acid and aromatic guanidino amino acids have been synthesized. They resulted active against Grampositive bacteria relative to eukaryotic cells and showed also an appreciable activity against staphylococcal biofilms. This last effect can be related to the recognized activity of SAMPs on dormant bacterial cells and can represent an important goal to overcome the extremely enhanced tolerance to antibiotics expressed by persistent bacteria responsible for the relapse in microbial biofilm infections.