OH-CATH

King cobra cathelicidin (OH-CATH) is composed of 34 amino acid residues having strong antibacterial and very weak hemolytic activities as reported by us recently. OH-CATH can be served as a valuable template to develop novel therapeutic drugs. In this study, OH-CATH and six of its analogs were synthesized to explore their structure-function relationships based on their bactericidal and hemolytic activities. Experimental results of OH-CATH(3-34) and OH-CATH(5-34) indicated that the N-terminal 4 amino acid residues of OH-CATH played an important role on its hemolytic activity but had weak effects on its bactericidal activity. Among OH-CATH and its analogs, OH-CATH(5-34) had the lowest hemolytic activity while maintained strong antimicrobial activity. To evaluate its potential usage, the biological activities of OH-CATH(5-34) were compared with those of pexiganan. The bactericidal activity of OH-CATH(5-34) against 5 different species (11 laboratory strains) was 2-4 times stronger than that of pexiganan (4-16 microg/ml vs 8-32 microg/ml). Hemolytic activity of OH-CATH(5-34) against human erythrocytes was 0.69% while that of pexiganan was 16.5% at the dosage of 200 microg/ml. OH-CATH(5-34) showed very weak cytotoxic activities against primary rabbit ventricular endothelial cells and four human cancer cell lines whereas pexiganan showed strong cytotoxic activity against these five cell lines (IC(50)=20-90 microg/ml). The intravenous LD(50) value of OH-CATH(5-34) on mice was 7-fold higher than that of pexiganan (175 mg/kg vs 25mg/kg). Taken together, our results suggested that OH-CATH(5-34) should be considered as an excellent candidate for developing therapeutic drugs.

Cationic antimicrobial peptides (AMPs) are considered as important candidate therapeutic agents, which exert potent microbicidal properties against bacteria, fungi and some viruses. Based on our previous findings king cobra cathelicidin (OH-CATH) is a 34-amino acid peptide that exerts strong antibacterial and weak hemolytic activity. The aim of this research is to evaluate the efficacy of both OH-CATH30 and its analog D-OH-CATH30 against clinical isolates comparing with routinely utilized antibiotics in vitro. In this study, 584 clinical isolates were tested (spanning 2013-2016) and the efficacy of the candidate peptides and antibiotics were determined by a broth microdilution method according to the CLSI guidelines. Among the 584 clinical isolates, 85% were susceptible to OH-CATH30 and its analogs. Both L- and D-OH-CATH30 showed higher efficacy against (toward) Gram-positive bacteria and stronger antibacterial activity against nearly all Gram-negative bacteria tested compare with antibiotics. The highest bactericidal activity was detected against Acinetobacter spp., including multi-drug-resistant Acinetobacter baumannii (MRAB) and methicillin-resistant Staphylococcus aureus (MRSA). The overall efficacy of OH-CATH30 and its analogs was higher than that of the 9 routinely used antibiotics. OH-CATH30 is a promising candidate drug for the treatment of a wide variety of bacterial infections which are resistant to many routinely used antimicrobial agents.

To investigate the potential protective effects of the snake venom antimicrobial peptide OH-CATH, we used a series of rabbit urinary tract infection models successfully induced by cephalosporin-resistant E.coli and E. coli ATCC 25922. The experimental models were administered saline, snake venom antimicrobial peptide OH-CATH, Cefoperazone and Sulbactam through the urethra. Urine was collected on days 1, 5, 10 and 14 after model establishment and urine culture was done to check the infection in each experimental animals. On day 14, all the animals were sacrificed and the bladder tissue specimens were taken for observation by H-E staining light microscope and transmission electron microscope. We found that the snake venom antimicrobial peptide OH-CATH reduced bacterial count in urine culture in both cephalosporin-resistant E. coli and the E. coli ATCC 25922 infected animals, while Cefoperazone and Sulbactam were only able to reduce the positive rate induced by the E. coli ATCC 25922 but had no obvious effects on animal model induced by cephalosporin-resistant E. coli strains (P<0.05). We also found less necrosis, degeneration and inflammatory cell infiltration in bladder tissue in OH-CATH groups as compared with the other experimental groups. The snake venom antimicrobial peptide OH-CATH had stable antibacterial activity against cephalosporin-resistant E. coli and E. coli ATCC 25922 and exhibited protective effects on both the cephalosporin-resistant E. coli and E. coli ATCC 25922 rabbit urinary tract infection models, suggesting that the molecule may have potential clinical applications in treating urinary tract infections.