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EP-4737573-A1 - POLYPEPTIDE PERMEABLE THROUGH OUTER MEMBRANE OF GRAM-NEGATIVE BACTERIUM, ANTIBACTERIAL PROTEIN COMPRISING SAID POLYPEPTIDE, ANTIBACTERIAL AGENT OR DISINFECTANT, PHARMACEUTICAL COMPOSITION, AND ANTIBACTERIAL AGENT COMPOSITION OR DISINFECTANT COMPOSITION

EP4737573A1EP 4737573 A1EP4737573 A1EP 4737573A1EP-4737573-A1

Abstract

The present invention provides a polypeptide as defined in one of (i) to (iii) below, as well as an antibacterial protein, an antibacterial agent, a disinfectant, a pharmaceutical composition, an antibacterial agent composition, and a disinfectant composition, each comprising the following polypeptide: (i) a polypeptide comprising the amino acid sequence represented by SEQ ID NO: 1; (ii) a polypeptide comprising an amino acid sequence in which one or more amino acid residues are deleted, substituted, inserted, and/or added in the amino acid sequence represented by SEQ ID NO: 1; or (iii) a polypeptide comprising an amino acid sequence having sequence identity of 80% or higher with the amino acid sequence represented by SEQ ID NO: 1.

Inventors

  • UCHIYAMA, Jumpei
  • UCHIYAMA, Iyo
  • FUKUDA, KEN
  • YAMASHIRO, KENJI

Assignees

  • National University Corporation Okayama University
  • National University Corporation Kochi University

Dates

Publication Date
20260506
Application Date
20240628

Claims (12)

  1. A polypeptide capable of permeating a lipopolysaccharide-containing outer membrane of Gram-negative bacteria, as defined in one of (i) to (iii) below: (i) a polypeptide comprising the amino acid sequence represented by SEQ ID NO: 1; (ii) a polypeptide comprising an amino acid sequence in which one or more amino acid residues are deleted, substituted, inserted, and/or added in the amino acid sequence represented by SEQ ID NO: 1; or (iii) a polypeptide comprising an amino acid sequence having sequence identity of 80% or higher with the amino acid sequence represented by SEQ ID NO: 1.
  2. A polynucleotide as defined in one of (iv) to (vi) below: (iv) a polynucleotide comprising a DNA sequence encoding the polypeptide according to claim 1; (v) a polynucleotide comprising the DNA sequence represented by SEQ ID NO: 2; or (vi) a polynucleotide comprising a DNA sequence having sequence identity of 80% or higher with the polynucleotide comprising a DNA sequence as defined in (iv) or (v), and encoding a polypeptide capable of permeating a lipopolysaccharide-containing outer membrane of Gram-negative bacteria.
  3. An antibacterial protein comprising an endolysin and the polypeptide according to claim 1, wherein the polypeptide according to claim 1 is located at the N-terminus or C-terminus of the endolysin.
  4. A polynucleotide encoding the antibacterial protein according to claim 3.
  5. An antibacterial agent or disinfectant against Gram-negative bacteria, comprising the antibacterial protein according to claim 3.
  6. The antibacterial agent or disinfectant according to claim 5, wherein the Gram-negative bacteria are multidrug-resistant bacteria or Pseudomonas aeruginosa.
  7. A pharmaceutical composition comprising the antibacterial protein according to claim 3.
  8. The pharmaceutical composition according to claim 7, wherein a disease caused by the Pseudomonas aeruginosa is one or more diseases selected from the group consisting of keratitis, dacryocystitis, endophthalmitis, cellulitis-induced eye infections, sepsis, pneumonia, multidrug-resistant Gram-negative bacterial infections, opportunistic infections, and other Pseudomonas aeruginosa-induced diseases.
  9. An antibacterial composition or disinfectant composition comprising: the antibacterial protein according to claim 3; and an outer membrane permeabilizer, wherein the outer membrane permeabilizer is ethylenediaminetetraacetic acid (EDTA) or a salt thereof.
  10. The antibacterial composition or disinfectant composition according to claim 9, wherein the Gram-negative bacteria are multidrug-resistant bacteria or Pseudomonas aeruginosa.
  11. A method for preventing a disease caused by a Gram-negative bacterial infection or for killing Gram-negative bacteria, the method comprising administering a pharmaceutical composition containing the antibacterial protein according to claim 3 to a human.
  12. A method for preventing a disease caused by a Gram-negative bacterial infection or for killing Gram-negative bacteria, the method comprising administering a pharmaceutical composition containing the antibacterial protein according to claim 3 to an animal.

Description

Technical Field The present invention relates to a polypeptide capable of permeating an outer membrane of Gram-negative bacteria, as well as an antibacterial protein, an antibacterial agent or disinfectant, a pharmaceutical composition, and an antibacterial composition or disinfectant composition, each containing the polypeptide. In the present specification, the "polypeptide capable of permeating a lipopolysaccharide-containing outer membrane of Gram-negative bacteria" is sometimes abbreviated as "outer membrane-permeable polypeptide." Background Art Pseudomonas aeruginosa is a pathogenic bacterium that is inherently drug-resistant and that acquires drug resistance, causing intractable infections in humans and animals (such as cattle and dogs). The bacterium is also included in WHO's list of drug-resistant bacteria of high emergency importance. Today, there is a need to develop a novel antibacterial agent against Pseudomonas aeruginosa. Investigation on conventional antibiotics, which are low-molecular-weight compounds, is continuing due to the presence of established development frameworks. However, the library of low-molecular-weight compounds is limited, making it very difficult to discover anti-infective drugs with novel mechanisms of action. Given this situation, antimicrobial peptides, phages, and endolysins are currently being investigated as entirely different and novel modalities for anti-infective drugs. Endolysins have been investigated, with the focus on natural-phage endolysins. PTL 1 discloses an endolysin protein targeting staphylococci. NPL 1 to NPL 3 each disclose an endolysin protein against Gram-negative bacteria. Basically, the cell wall of Gram-negative bacteria includes an outer membrane, and a natural endolysin alone cannot kill the bacteria. Meanwhile, a natural endolysin capable of killing such Gram-negative bacteria does not have the bactericidal effect necessary and sufficient for drugs to treat infectious diseases. Citation List Patent Literature PTL 1: JP2021-112152A Non-patent Literature NPL 1: Lai WCB, Chen X, Ho MKY, Xia J, Leung SSY. Bacteriophage-derived endolysins to target Gram-negative bacteria. Int J Pharm. 589, 119833, 2020.NPL 2: De Maesschalck V, Gutierrez D, Paeshuyse J, Lavigne R, Briers Y. Advanced engineering of third-generation lysins and formulation strategies for clinical applications. Crit Rev Microbiol. 46(5), 548-564, 2020.NPL 3: Gontijo MTP, Jorge GP, Brocchi M. Current Status of Endolysin-Based Treatments against Gram-negative Bacteria. Antibiotics (Basel). 10(10), 1143, 2021. Summary of Invention Technical Problem One aim of the present invention is to provide an antibacterial agent or disinfectant having efficacy against Gram-negative bacteria, including multidrug-resistant bacteria, particularly Pseudomonas aeruginosa. Another aim of the present invention is to provide a polypeptide capable of increasing the outer membrane permeability of Gram-negative bacteria to endolysin, and a polynucleotide encoding the polypeptide. Solution to Problem The present invention provides an outer membrane-permeable polypeptide, an antibacterial protein, a polynucleotide encoding the polypeptide and the protein, an antibacterial agent or disinfectant, a pharmaceutical composition, and an antibacterial composition or disinfectant composition. (1) A polypeptide capable of permeating a lipopolysaccharide-containing outer membrane of Gram-negative bacteria, as defined in one of (i) to (iii) below: (i) a polypeptide comprising the amino acid sequence represented by SEQ ID NO: 1; (ii) a polypeptide comprising an amino acid sequence in which one or more amino acid residues are deleted, substituted, inserted, and/or added in the amino acid sequence represented by SEQ ID NO: 1, and capable of permeating a lipopolysaccharide-containing outer membrane of Gram-negative bacteria; or(iii) a polypeptide comprising an amino acid sequence having sequence identity of 80% or higher with the amino acid sequence represented by SEQ ID NO: 1, and capable of permeating a lipopolysaccharide-containing outer membrane of Gram-negative bacteria.(2) A polynucleotide as defined in one of (iv) to (vi) below: (iv) a polynucleotide comprising a DNA sequence encoding the polypeptide according to (1);(v) a polynucleotide comprising the DNA sequence represented by SEQ ID NO: 2: or(vi) a polynucleotide comprising a DNA sequence having sequence identity of 80% or higher with the polynucleotide comprising a DNA sequence as defined in (iv) or (v), and encoding a polypeptide capable of permeating a lipopolysaccharide-containing outer membrane of Gram-negative bacteria.(3) An antibacterial protein comprising an endolysin and the polypeptide according to (1), wherein the polypeptide according to (1) is located at the N-terminus or C-terminus of the endolysin.(4) A polynucleotide encoding the antibacterial protein according to (3).(5) An antibacterial agent or disinfectant against Gram-negative bacteria, compri