Liposomal entrapment of the neutrophil-derived peptide indolicidin endows it with in vivo antifungal activity

Abstract

Indolicidin, a cationic tridecapeptide amide isolated from the granules of bovine neutrophils, has been found to possess potent antimicrobial activity in vitro but its nonselective toxicity could restrict its therapeutic utility. We found that the concentration at which indolicidin disrupts washed human red blood cell membranes coincided with the concentration at which indolicidin self associates. Because of a preponderance of hydrophobic residues, we believed that indolicidin would partition into liposomes which would restrict its exchange with biological tissues and consequently reduce its toxicity. Fluorescence spectroscopy of indolicidin added to 100 nm liposomes comprised of POPC, POPC/cholesterol (60:40 mol%), DPPC, or DPPC/cholesterol (60:40) revealed a large blue-shift and an increase in intensity of the emission profile indicating insertion into the bilayer. Of the lipids tested, POPC exhibited the highest degree of indolicidin binding as determined by fluorescence and encapsulation efficiency. By sequestering indolicidin within the lipid bilayer of 100 nm POPC liposomes we significantly reduced its toxicity to CHO/K1 cells. Likewise, the systemic toxicity of liposomal indolicidin in Balb/c mice was decreased dramatically relative to aqueous solutions; the maximum dose at which no deaths occurred was 0.4 mg/kg for free indolicidin versus 40 mg/kg for indolicidin-POPC. Because of this decrease in toxicity, we were able to administer liposomally encapsulated material at significantly higher concentrations than unencapsulated aqueous material and achieve efficacy in treating animals systemically infected with Aspergillus fumigatus. Liposomal but not free indolicidin was found to be effective in obtaining cures. This report is the first description of the in vivo therapeutic activity of a neutrophil-derived antimicrobial peptide and suggests that liposomal treatment modalities will provide effective strategies for endowing this class of compounds with pharmacological utility.

Pharmacodynamic Behavior of Liposomal Antisense Oligonucleotides Targeting Her-2/neu and Vascular Endothelial Growth Factor in an Ascitic MDA435/LCC6 Human Breast Cancer Model

“…As is true for drugs such as anti-fungals, 37 anti-bacterials 38 and several anticancer agents, [39][40][41][42] various ASO are being developed as liposomal formulations in order to protect the therapeutic agent from degradation. [43][44][45] A commonly used modification of ASO is the chemical variation of the backbone, the most commonly used being that of the substitution of a sulfur atom for one of the nonbridging oxygen atoms in the phosphodiester bond.…”

Determination of the Secondary Structure and Conformation of Puroindolines by Infrared and Raman Spectroscopy

“…Ahmad et al (1995) have observed that indolicidin, a FIGURE 9: Comparison of the predicted R-helices for puroindoline-a and puroindoline-b. Helices were predicted from the sequence homology (Align.)…”

Porcine antimicrobial peptides: New prospects for ancient molecules of host defense

“…Structure prediction analysis and circular dichroism spectra suggest that PMAP-36, and -37 adopt an amphipathic α-helix, whereas PMAP-23 assumes a hairpin-like structure (an antiparallel β-sheet connected by a loop at center). Sequence comparison reveals that PMAP-23 shows no significant similarity to any known AMPs, whereas PMAP-36 (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) has a moderate homology of 35% to a rabbit cathelicidin CAP18. Although PMAP-37 shows negligible identity to porcine cecropin P1, it has more than 50% similarity to two insect AMPs, cecropins A and B.…”

“…Prominent induction of genes for many epithelial β-defensins occurs during skin injuries and airway and enteric infections [98,111,117]. In addition, administration of AMPs to mice has been shown to provide significant protection against lethal endotoxic shock or experimental infections [3,45,107].…”