Klebsiella is a common Gram-negative pathogen causing community-acquired bacterial pneumonia, and K. pneumoniae pneumonia is considered the most common cause of hospital-acquired pneumonia in the United States. K. pneumoniae is a difficult infection to treat because of the organism's thick capsule that is usually best treated with the last line antibiotic therapy such as carbapenems. However, carbapenem-resistant K. pneumoniae (CRKP), one of the carbapenem-resistant Enterobacteriaceae (CRE), is an emerging cause of antibiotic-resistant nosocomial infections associated with high rates of morbidity and mortality. New therapies in controlling CRKP-induced infections are urgently needed. Using recombinant protein and genetically modified mouse knockout models, we have demonstrated that the short palate, lung, nasal epithelium clone (PLUNC) 1 (SPLUNC1) contributes to pulmonary host defense against K. pneumoniae induced infection. A novel SPLUNC1- derived peptide from the antimicrobial motif of the SPLUNC1 protein, a4-Short, demonstrated more potent antibacterial properties than the full-length recombinant SPLUNC1 protein and in vivo efficacy in a murine model of respiratory infection. Based on additional modifications of a4-Short, we have recently developed a series of rationally engineered antimicrobial peptides (AMPs) that rapidly kill their microbial targets by permeabilizing bacterial membrane regardless of the specific metabolic state of the bacteria. One of our lead AMPs, A4-153, has demonstrated potent bactericidal activity against diverse difficult-to-treat multidrug resistant (MDR) pathogens, including CRKP. Exciting, A4-153 is active against many CRKP that have developed resistance to other membrane-active compounds, such as the natural AMP LL37 and colistin, an antibiotic of last resort. In addition, we have found a substantially lower tendency for bacteria to develop resistance to A4-153 compared to standard antibiotic agents and natural AMPs. Importantly, we found that similar to natural AMP LL37, A4-153 displayed no detectable hemolysis and could be safely administered to mouse lungs with very high concentrations. We propose in this SBIR application to explore the feasibility of using the newly developed A4- 153 to prevent CRKP-induced pneumonia by killing CRKP and eradicating the CRKP biofilm in the abiotic and biotic system using in vitro and in vivo models. The successful completion of the proposed aims in this Phase I application will prepare us for IND-enabling studies to be presented in a subsequent Phase II application, including multidose MTD, GLP toxicity in rodents and large animals, and initiation of GMP manufacturing.
Public Health Relevance Statement: Project Narrative/Public Health Relevance Statement The increase in respiratory infection with multidrug-resistant bacteria, including carbapenem-resistant Klebsiella pneumoniae (CRKP), worsens the difficulty of effectively treating bacterial pathogens using stand-of-care antibiotics. New therapies in controlling CRKP-induced infections are urgently needed. This Phase I SBIR project aims to explore the feasibility of using the newly developed antimicrobials in treating CRKP-induced pneumonia using biologically relevant models. Successful completion of this proposed project will identify a novel class of peptide-based antibiotics to alleviate CRKP infection-associated healthcare problems.
Project Terms: K pneumoniae; K. pneumoniae; Klebsiella pneumoniae; Air; Animals; Antibiotic Agents; Antibiotic Drugs; Miscellaneous Antibiotic; Antibiotics; Bacteria; Bacterial Infections; bacteria infection; bacterial disease; Bacterial Pneumonia; bacteria pneumonia; Blood; Blood Reticuloendothelial System; capsule; Capsules; Carbapenems; Cell Survival; Cell Viability; Colistin; Colimycin; Colisticin; Polymyxin E; Communities; Engineering; Epithelial Cells; Exhibits; Half-Life; Hemolysis; erythrolysis; In Vitro; indexing; Infection; Kinetics; Klebsiella; Calymmatobacterium; Donovania; Lead; Pb element; heavy metal Pb; heavy metal lead; Lung; Lung Respiratory System; pulmonary; Morbidity - disease rate; Morbidity; mortality; Mus; Mice; Mice Mammals; Murine; Organism; living system; Peptides; Permeability; Drug Kinetics; Pharmacokinetics; Pneumonia; Proteins; Public Health; Recombinant Proteins; Respiratory Tract Infections; Airway infections; Respiratory Infections; Rodent; Rodentia; Rodents Mammals; Safety; Septicemia; Blood Poisoning; septicaemia; septicemic; Testing; Tissues; Body Tissues; Treatment Protocols; Treatment Regimen; Treatment Schedule; United States; Urine; Measures; Microbial Biofilms; biofilm; Resistance to antibiotics; Resistant to antibiotics; antibiotic drug resistance; antibiotic resistant; Antibiotic Resistance; health care; Healthcare; Caring; Blood Sample; Blood specimen; Clinical; Phase; Physiologic; Physiological; Series; Hospital Infections; Hospital acquired infection; institutional infection; Nosocomial Infections; KO mice; Knock-out Mice; Null Mouse; Knockout Mice; Susceptibility; Predisposition; Blood Serum; Serum; Multidrug Resistance; Multiple Drug Resistance; Multiple Drug Resistant; Resistance to Multi-drug; Resistance to Multidrug; Resistance to Multiple Drug; Resistant to Multiple Drug; Resistant to multi-drug; Resistant to multidrug; multi-drug resistant; multidrug resistant; Multi-Drug Resistance; Antibacterial Agents; anti-bacterial; antibacterial; Anti-Bacterial Agents; beta lactam antibiotic; ß lactam antibiotic; ß-Lactams; beta-Lactams; Therapeutic; fluid; liquid; Liquid substance; Metabolic; Antibiotic Therapy; Antibiotic Treatment; bacterial disease treatment; bacterial infectious disease treatment; fighting; Minimum Inhibitory Concentration measurement; Minimum Inhibitory Concentrations; Frequencies; Protocols documentation; Protocol; System; meter; Host Defense; cytotoxicity; Lytotoxicity; Membrane; membrane structure; beta-Lactamase; G24 protein; beta lactam hydrolase; beta-Lactamhydrolase; ß-Lactamase; Nasal Epithelium; microbial; Toxic effect; Toxicities; novel; Palate; Maximum Tolerated Dose; Maximal Tolerated Dose; Maximally Tolerated Dose; Modeling; Property; Nosocomial pneumonia; healthcare-associated pneumonia; hospital acquired pneumonia; hospital associated pneumonia; Bacterial Antibiotic Resistance; antibiotic resistant bacteria; bacterial antibiotic resistant; bacterial resistance to antibiotic; Multiple Bacterial Drug Resistance; Multiple Anti-bacterial Drug Resistance; Multiple Anti-bacterial Drug Resistant; Multiple Antibacterial Drug Resistance; Multiple Antibacterial Drug Resistant; Resistance to Multiple Anti-bacterial Drug; Resistance to Multiple Antibacterial Drug; Resistant to Multiple Anti-bacterial Drug; Resistant to Multiple Antibacterial Drug; multi-drug resistant bacteria; multidrug resistant bacteria; Pathogenicity; anti-microbial peptide LL-37; antimicrobial peptide LL-37; Thickness; Thick; Tissue Sample; preventing; prevent; Address; Length; Dose; global health; Recombinants; in vivo; in vivo Model; Collection; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Modification; antimicrobial peptide; anti-microbial peptide; bactericide; bactericidal; next generation; blood infection; bloodstream infection; Sepsis; Resistant development; developing resistance; Resistance development; pathogen; bacteria pathogen; bacterial pathogen; pathogenic bacteria; anti-microbial; antimicrobial; Respiratory Epithelium; Structure of respiratory epithelium; respiratory tract epithelium; airway epithelium; natural antimicrobial; new drug treatments; new drugs; new pharmacological therapeutic; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel pharmaco-therapeutic; novel pharmacological therapeutic; novel therapy; novel therapeutics; murine model; mouse model; develop therapy; intervention development; treatment development; therapy development; public health relevance; carbapenem resistant; resistance to carbapenem; resistant to carbapenem; carbapenem resistance; MDR organism; MDR pathogen; multi-drug resistant organism; multidrug resistant organism; multidrug resistant pathogen; multiple drug resistant organism; multiple drug resistant pathogen; multi-drug resistant pathogen; carbapenem-resistant Enterobacteriaceae; extensively drug resistant; extreme drug resistance; extensive drug resistance; therapeutic candidate; health care-associated infections; healthcare-associated infections; lead optimization; antibiotic resistant pathogen; drug resistant pathogen; optimal therapies; optimal treatments; pneumonia models; pneumonia model; platelet function; resistance in K pneumoniae; resistance in K. pneumoniae; resistance in Klebsiella pneumoniae; resistant K pneumoniae; resistant K. pneumoniae; resistant Klebsiella pneumoniae; antibiotic resistant infections; manufacture
