Efficiency of NZ2114 on Superficial Pyoderma Infected with Staphylococcus pseudintermedius

Abstract

Staphylococcus pseudintermedius (S. pseudintermedius) is the main pathogen causing pyoderma of canines. With the emergence of drug-resistant bacteria, traditional antibiotic treatments are limited. As a potential antibacterial agent, NZ2114 was effective against S. pseudintermedius, including drug-resistant strains. Its bactericidal efficacy was superior to mupiroxacin, ofloxacin and lincomycin. To facilitate the transcutaneous delivery of NZ2114 for the treatment of superficial pyoderma, chemical permeation enhancers were added since water-soluble NZ2114 does not easily penetrate the skin lipid layer. Two different NZ2114 sprays were prepared by combining 1% Azone + 10% propylene glycol (PG) or 5% N-methylpyrrolidone (NMP) + 10% PG with NZ2114 after screening. The cumulative permeability of NZ2114 sprays were 244.149 and 405.245 μg/cm2 at 24 h with an in vitro percutaneous assay of mice skin, which showed a 244% and 405% increase in skin permeability than NZ2114, respectively. In addition, the efficacy of NZ2114 sprays in reducing skin bacteria colonisation was demonstrated in a mouse model of superficial pyoderma (24 mice, 3 mice/group) induced by S. pseudintermedius, and the 5% NMP + 10% PG + NZ2114 group had the best therapeutic effect compared to the other groups. This preparation did not cause any skin irritation, laying the foundation for the development of an effective and non-toxic topical product.

In Vitro/Vivo Mechanisms of Antibacterial Peptide NZ2114 against Staphylococcus pseudintermedius and Its Biofilms

“…In previous studies, NZ2114 displayed bactericidal activity with high efficiency towards Gram-positive bacteria, including S. suis, S. aureus, S. dysgalactiae, and Clostridium perfringens [15,21,22]. In this study, NZ2114 displayed more potent antibacterial activity towards S. pseudintermedius than those of mupirocin (MIC: 0.25-0.5 µM) and lincomycin (MIC: 4.34-69.41 µM) (Table 1) similar to previous observation [19]. Jarosiewicz et al evaluated the antimicrobial activity of seven AMPs (aurein 1.2, CAMEL, citropin 1.1, protegrin-1, pexiganan, temporin A, and uperin 3.6) against fifty-three methicillin-sensitive S. pseudintermedius (MSSP) and seven methicillin-resistant S. pseudintermedius (MRSP); all tested peptides were active against all reference-and clinical strains, in which uperin 3.6 showed the better antimicrobial activity (MIC 90 = 2 µg/mL) [20].…”

“…The in vivo therapy of NZ2114 in a mouse pyoderma model induced by S. pseudintermedius showed that it effectively decreased the number of skin bacteria, alleviated histological damage, and reduced the skin damage area. These results, including efficacy results on preparation optimization together [19], fully demonstrated that NZ2114 can potentially be used in the treatment of S. pseudintermedius infections.…”

“…NZ2114, moreover, a derived peptide of plectasin, exhibited better antibacterial activity toward S. aureus with low hemolytic activity [17], and NZ2114 also effectively inhibited the biofilm-forming ability of Staphylococcus dysgalactiae and S. aureus, eradicating biofilm [15,18]. Therefore, NZ2114 is a promising antimicrobial that can inhibit and eradicate S. pseudintermedius and its biofilms, and the former effect was reported in early work [19]. In addition, the heterologous expression of NZ2114 was successfully established by a high-yield Pichia pastoris expression system [17], which is a step towards a cost-effective means of development.…”

“…What is more important is that an AMP fermentation and purification platform system with an industrial scale of 20 m 3 and 30 m 3 was established in China in 2019 and 2021, respectively [15], which broke the bottleneck of AMP industrialization. However, no systemic studies in vitro and vivo have focused on the process details, mechanisms and mode of action of NZ2114 against S. pseudintermedius and biofilm so far except for our previous partial preliminary observation [19].…”