BACKGROUND: Among the non-traditional antibacterial agents in development, only a few targets critical Gram-negative bacteria such as carbapenem-resistant Pseudomonas aeruginosa, Acinetobacter baumannii or cephalosporin-resistant Enterobacteriaceae. Endolysins and their genetically modified versions meet the World Health Organization criteria for innovation, have a novel mode of antibacterial action, no known bacterial cross-resistance, and are being intensively studied for application against Gram-negative pathogens. METHODS: The study presents a multidisciplinary approach, including genetic engineering of LysECD7-SMAP and production of recombinant endolysin, its analysis by crystal structure solution following molecular dynamics simulations and evaluation of antibacterial properties. Two types of antimicrobial dosage forms were formulated, resulting in lyophilized powder for injection and hydroxyethylcellulose gel for topical administration. Their efficacy was estimated in the treatment of sepsis, and pneumonia models in BALB/c mice, diabetes-associated wound infection in the leptin receptor-deficient db/db mice and infected burn wounds in rats. RESULTS: In this work, we investigate the application strategies of the engineered endolysin LysECD7-SMAP and its dosage forms evaluated in preclinical studies. The catalytic domain of the enzyme shares the conserved structure of endopeptidases containing a putative antimicrobial peptide at the C-terminus of polypeptide chain. The activity of endolysins has been demonstrated against a range of pathogens, such as Klebsiella pneumoniae, A. baumannii, P. aeruginosa, Staphylococcus haemolyticus, Achromobacter spp, Burkholderia cepacia complex and Haemophylus influenzae, including those with multidrug resistance. The efficacy of candidate dosage forms has been confirmed in in vivo studies. Some aspects of the interaction of LysECD7-SMAP with cell wall molecular targets are also discussed. CONCLUSIONS: Our studies demonstrate the potential of LysECD7-SMAP therapeutics for the systemic or topical treatment of infectious diseases caused by susceptible Gram-negative bacterial species and are critical to proceed LysECD7-SMAP-based antimicrobials trials to advanced stages.

• Klíčová slova
• Dosage forms, Engineered endolysin, Enzyme-based antibacterial, Gram-negative bacteria, Preclinical efficacy,
• MeSH
• antibakteriální látky farmakologie   aplikace a dávkování   MeSH
• endopeptidasy * farmakologie   aplikace a dávkování   MeSH
• gramnegativní bakteriální infekce * farmakoterapie   MeSH
• gramnegativní bakterie * účinky léků   MeSH
• krysa rodu Rattus MeSH
• myši inbrední BALB C * MeSH
• myši MeSH
• proteinové inženýrství metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• endolysin MeSH Prohlížeč
• endopeptidasy * MeSH

The effect of combined proteolytic enzymes, administered by the rectal route, on the metastatic process and the time of survival in C57Bl6 mice with the Lewis lung carcinoma inoculated subcutaneously was investigated. In the control group, which received no enzyme treatment, 90% of animals died of the metastatic spread of cancer by day 18 after primary tumor extirpation. In Group A, which received the multi-enzyme solution from the time of primary tumor extirpation, 30% of mice died of disseminated cancer by day 25. In Group B, which was treated with the enzymes from 6 days before primary tumor extirpation, only 10% of animals showed the metastatic process by day 15. In Group C, which received the enzymes from 24 hours after intracutaneous tumor inoculation, no metastatic dissemination was discernible. In these three groups, the enzyme treatment was carried out throughout the study. None of the control animals survived for 100 days when the study was ended. The treated groups A, B and C showed survival rate 60%, 90% and 100% of animals, respectively, by 100 days.

• MeSH
• aplikace rektální MeSH
• chymotrypsin * MeSH
• endopeptidasy aplikace a dávkování   farmakologie   MeSH
• extrakty z thymu aplikace a dávkování   farmakologie   MeSH
• fixní kombinace léků MeSH
• karcinom plic Lewisové mortalita   patologie   prevence a kontrola   MeSH
• metastázy nádorů patologie   prevence a kontrola   MeSH
• míra přežití MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory kůže mortalita   patologie   prevence a kontrola   MeSH
• pankreatické extrakty aplikace a dávkování   farmakologie   MeSH
• papain aplikace a dávkování   farmakologie   MeSH
• transplantace nádorů MeSH
• trypsin * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chymotrypsin, papain, trypsin drug combination MeSH Prohlížeč
• chymotrypsin * MeSH
• endopeptidasy MeSH
• extrakty z thymu MeSH
• fixní kombinace léků MeSH
• pankreatické extrakty MeSH
• papain MeSH
• trypsin * MeSH

• MeSH
• aktivátor plazminogenu urokinázového typu aplikace a dávkování   MeSH
• dospělí MeSH
• endopeptidasy aplikace a dávkování   MeSH
• injekce MeSH
• krvácení farmakoterapie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• oční nemoci farmakoterapie   MeSH
• senioři MeSH
• sklivec * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• aktivátor plazminogenu urokinázového typu MeSH
• endopeptidasy MeSH