Modification of a novel angiogenic peptide, AG30, for the development of novel therapeutic agents

Abstract

We previously identified a novel angiogenic peptide, AG30, with antibacterial effects that could serve as a foundation molecule for the design of wound-healing drugs. Toward clinical application, in this study we have developed a modified version of the AG30 peptide characterized by improved antibacterial and angiogenic action, thus establishing a lead compound for a feasibility study. Because AG30 has an α-helix structure with a number of hydrophobic and cationic amino acids, we designed a modified AG30 peptide by replacing several of the amino acids. The replacement of cationic amino acids (yielding a new molecule, AG30/5C), but not hydrophobic amino acids, increased both the angiogenic and the antimicrobial properties of the peptide. AG30/5C was also effective against methicillin-resistant Staphylococcus aureus (MRSA) and antibiotic-resistant Pseudomonas aeruginosa. In a diabetic mouse wound-healing model, the topical application of AG30/5C accelerated wound healing with increased angiogenesis and attenuated MRSA infection. To facilitate the eventual clinical investigation/application of these compounds, we developed a large-scale procedure for the synthesis of AG30/5C that employed the conventional solution method and met Good Manufacturing Practice guidelines. In the evaluation of stability of this peptide in saline solution, RP-HPLC analysis revealed that AG30/5C was fairly stable under 5°C for 12 months. Therefore, we propose the use of AG30/5C as a wound-healing drug with antibacterial and angiogenic actions.

AJP001, a novel helper T‐cell epitope, induces a humoral immune response with activation of innate immunity when included in a peptide vaccine

“…We previously developed a novel functional peptide, AG30 that possesses both angiogenic and antibacterial functions . To move this peptide toward clinical application, we evaluated several candidate peptides by modifying AG30 to enhance its function and stability.…”

“…We previously developed a novel functional peptide, AG30 that possesses both angiogenic and antibacterial functions. [9][10][11] To move this peptide toward clinical application, we evaluated several candidate peptides by modifying AG30 to enhance its function and stability. Through the process of peptide screening, the skin sensitization potency of these peptides was evaluated by the human Cell Line Activation Test (h-CLAT), which is a toxicity screening method that measures CD86 and CD54 expression in THP-1 cells.…”

“…5,6 Alum is a wellknown adjuvant that drives a Th2-biased immune response and induces the release of endogenous danger signals, also called "alarmins", via localized cellular damage, 7 and these alarmins directly stimulate inflammasomes via NLRP3. 8 We previously developed an antimicrobial peptide, termed angiogenic peptide 30 (AG30), with a length of 30 amino acids that possesses both angiogenic and antibacterial functions [9][10][11] similar to the functions of LL-37 and PR39. 12,13 We further designed and synthesized a series of AG30 analogs and identified a candidate adjuvant peptide (AJP001), which strongly induced the activation of inflammasomes and the NF-κB pathway.…”

Investigator‐initiated clinical study of a functional peptide, SR‐0379, for limb ulcers of patients with Werner syndrome as a pilot study

“…We have developed a number of novel peptide drugs to treat skin ulcers over the past 10 years, beginning with a discovery in basic science experiments and extending to clinical trials to evaluate the safety and efficacy for the treatment of severe limb ulcers. 8,13,[15][16][17] In the present study, we evaluated the effect of SR-0379 on limb ulcers in Werner syndrome patients.…”

Physician‐initiated first‐in‐human clinical study using a novel angiogenic peptide, AG30/5C, for patients with severe limb ulcers

“…In the evaluation of angiogenic activity, AG30/5C induced endothelial cell growth, migration and tube formation. Additionally, AG30/5C is effective in treating ulcers in mice . Interestingly, antibacterial effects of AG30/5C are observed against Escherichia coli , Pseudomonas aeruginosa and Staphylococcus aureus , as well as against various antibiotic‐resistant strains, such as methicillin‐resistant S. aureus (MRSA).…”