Antimicrobial agents

Inventors

Lavigne, Rob • Miller, Stefan • Briers, Yves • Volckaert, Guido • Walmagh, Maarten

Abstract

The application relates to antimicrobial agents against Gram-negative bacteria, in particular to fusion proteins composed of an enzyme having the activity of degrading the cell wall of Gram-negative bacteria and a peptide stretch fused to the enzyme at the N- or C-terminus, as well as pharmaceutical compositions comprising the same. Moreover, it relates to nucleic acid molecules encoding such a fusion protein, vectors comprising said nucleic acid molecules and host cells comprising either said nucleic acid molecules or said vectors. In addition, it relates to such a fusion protein for use as a medicament, in particular for the treatment or prevention of Gram-negative bacterial infections, as diagnostic means or as cosmetic substance. The application also relates to the treatment or prevention of Gram-negative bacterial contamination of foodstuff, of food processing equipment, of food processing plants, of surfaces coming into contact with foodstuff, of medical devices, of surfaces in hospitals and surgeries.

Core Innovation

The invention relates to a fusion protein having Gram-negative bacteria cell wall degrading activity, comprising an endolysin fused to a heterologous peptide segment. The peptide segment is fused to the endolysin at the N-terminus or C-terminus or at both termini, and exhibits an amino acid sequence selected from SEQ ID NO: 6 to 16 and 26 to 31. The resulting fusion protein is described as targeting Gram-negative bacteria by using the endolysin’s cell wall degrading activity together with the heterologous peptide segment.

The heterologous peptide segment is described in terms of sequence selection and peptide properties, including peptide segments that comprise amphipathic and/or cationic and/or hydrophobic characteristics as supported by the described peptide categories. The document further describes the fusion constructs as including peptide stretches exemplified by sushi peptides and defensin/AMP peptide stretches. The disclosed formats also include nucleic acids encoding the fusion proteins, and vectors and host cells for expressing the fusion proteins.

The document describes use cases where the fusion proteins are provided as medicaments and as disinfectant/sanitizing agents for controlling contamination. The disclosed applications include contamination control for food, surfaces, and medical devices, and the document also states diagnostic uses. The disclosure additionally provides supporting examples including fusion proteins with specific phage-derived and bacterial endolysins fused to selected peptide segments, and reported effects against Gram-negative strains.

Claims Coverage

The independent claim is clm-00001, which covers fusion proteins with Gram-negative bacterial cell wall degrading activity and specifies the terminal fusion arrangement plus selected peptide segment sequence ranges. Additional dependent claims refine the peptide segment and the claimed formats and uses, including peptide composition and length constraints, alternative sequence selections, and medicament and expression-related constructs.

Across the independent and dependent claims, the inventive coverage centers on endolysin-based fusion proteins for Gram-negative bacteria where a heterologous peptide segment is fused at the N- and/or C-terminus and is restricted to specified SEQ ID NO ranges, with additional claim refinements covering peptide composition, peptide length, alternative sequence selections, medicament use, and expression formats such as vectors.

Stated Advantages

Documented Applications