Haloganan: a novel antimicrobial peptide for treatmentof wound infections.

HG1 is a Leu-rich antimicrobial peptide (AMP). Previously, the peptide was shown to lose its activity in human serum although it possessed potent and broad spectrum antimicrobial activity against a wide range of pathogenic microbes. In an attempt to design an HG1 isomer that can overcome the problem of HG1, a structure­activity relationship study was conducted by substitution of each of five Leu residues with a Gln residue. Each substitute was tested for its antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) or Candida strains. In addition, the antimicrobial activity of HG1 isomers was examined in the presence of glycosaminoglycans or lipid components occurring in the extracellular matrix, human serum and wound fluid. As a result, it was determined that the third residue (Leu) in the sequence of HG1 was mainly responsible for abrogation of its antimicrobial activity in human serum or wound fluid. An HG1 isomer (L3Q) with a Gln-3 substitution exhibited a potent antibacterial activity in 50% human serum. While the anti-MRSA activity of L3Q was equivalent to that of HG1, its anti-Candida activity was found to be substantially reduced. In order to improve anti-Candida activity of L3Q, its cationicity was enhanced by replacement of the C-terminal Ala-19 with a Lys residue. Overall, an HG1 isomer with two substitutions of Gln-3 and Lys-19, named haloganan, was verified to have an advantage over HG1 in that it exerted its potent antimicrobial activity under conditions containing human serum and/or wound fluid.

Therapeutic efficacy of halocidin-derived peptide HG1 in a mouse model of Candida albicans oral infection.

OBJECTIVES: HG1 is an antimicrobial peptide derived from halocidin, which is naturally found in tunicates. The purpose of this study was to evaluate the therapeutic potential of HG1 as a novel antifungal agent for treating oral candidiasis. METHODS: The pharmacokinetic properties of HG1 were explored in mice, which were orally administered a single dose of HG1. Anti-Candida activity of HG1 was investigated in a time-dependent manner in the presence of saliva obtained from healthy donors or patients with oral candidiasis. In addition, HG1 was evaluated for its anti-Candida activity in the presence of proteins extracted from the culture supernatant of Candida albicans. The therapeutic potential in vivo and ex vivo of HG1 against oral candidiasis was investigated using a mouse model of oral candidiasis. RESULTS: Our data showed that absorption of HG1 into the blood did not occur following oral administration. In addition, HG1 exerted marked anti-Candida activity after short-term incubation at a concentration of 20 mg/L and it also caused a considerable reduction in fungal burden in the oral candidiasis mouse model when treated with 1 mg or 0.5 mg. CONCLUSIONS: This study suggests that HG1, as a novel component of mouthwash, might become an alternative antifungal agent to conventional drugs used to manage oral candidiasis.

Therapeutic efficacy of halocidin-derived peptide HG1 in a mouse model of surgical wound infection with methicillin-resistant Staphylococcus aureus.

We evaluated the therapeutic potential of HG1, an antimicrobial peptide, as a novel topical antibiotic by the use of a mouse surgical wound model of infection with methicillin-resistant Staphylococcus aureus. First, we attempted to determine whether or not HG1 infiltrated into the dermis when topically administered. Second, we evaluated the antibiotic effects of HG1 on skin infection via bacterial-enumeration and microscopic analyses. The results showed that topically administered HG1 was capable of penetrating into the dermis at the infection site, where it exerted its antimicrobial effects.

Antimicrobial activity of a halocidin-derived peptide resistant to attacks by proteases.

Cationic antimicrobial peptides (AMPs) have attracted a great deal of interest as a promising candidate for a novel class of antibiotics that might effectively treat recalcitrant infections caused by a variety of microbes that are resistant to currently available drugs. However, the AMPs are inherently limited in that they are inevitably susceptible to attacks by proteases generated by human and pathogenic microbes; this vulnerability severely hinders their pharmaceutical use in human therapeutic protocols. In this study, we report that a halocidin-derived AMP, designated HG1, was found to be resistant to proteolytic degradation. As a result of its unique structural features, HG1 proved capable of preserving its antimicrobial activity after incubation with trypsin, chymotrypsin, and human matrix metalloprotease 7 (MMP-7). Additionally, HG1 was observed to exhibit profound antimicrobial activity in the presence of fluid from human skin wounds or proteins extracted from the culture supernatants of Staphylococcus aureus and Pseudomonas aeruginosa. Greater understanding of the structural motifs of HG1 required for its protease resistance might provide feasible ways to solve the problems intrinsic to the development of an AMP-based antibiotic.