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JP-7856871-B2 - Klebsiella control

JP7856871B2JP 7856871 B2JP7856871 B2JP 7856871B2JP-7856871-B2

Inventors

  • デンコヴスキエネ エルナ
  • ミシウナス アウドリウス
  • ラザンスキエネ アウスラ

Assignees

  • ノマド バイオサイエンス ゲゼルシャフト ミット ベシュレンクテル ハフツング

Dates

Publication Date
20260512
Application Date
20200605
Priority Date
20190606

Claims (13)

  1. A pharmaceutical composition for use in a method of treating a target infection caused by Klebsiella, comprising a protein having cytotoxic activity against Klebsiella, wherein the protein comprises or consists of a first amino acid sequence segment and a second amino acid sequence segment directly bound in this order . (A) The first amino acid sequence segment consists of amino acid residues 1-127 of (A-ii) Sequence ID No. 2 (KvarM); Alternatively, (B) the first amino acid sequence segment consists of an amino acid sequence having at least 90% sequence identity with the amino acid sequence of amino acid residues 1 to 127 of (B-ii) Sequence ID No. 2; Alternatively, (C) the first amino acid sequence segment consists of an amino acid sequence having 1 to 10 amino acid substitutions, additions, insertions and/or deletions compared to the amino acid sequence of amino acid residues 1 to 127 of (C-ii) SEQ ID NO: 2, and (D) the second amino acid sequence segment consists of the amino acid sequence of amino acid residues 128 to 276 of (D-ii) SEQ ID NO: 2 (KvarM); Alternatively, (E) the second amino acid sequence segment consists of an amino acid sequence having at least 90% sequence identity with the amino acid sequence of amino acid residues 128-276 of (E-ii) Sequence ID No. 2; Alternatively, (F) the second amino acid sequence segment consists of an amino acid sequence having 1 to 15 amino acid substitutions, additions, insertions, or deletions compared to the amino acid sequence of amino acid residues 128 to 276 of (F-ii) SEQ ID NO : 2. Pharmaceutical composition.
  2. In item (B), the sequence identity is at least 95%; and/or in item (C), the number of amino acid substitutions, additions, insertions and/or deletions is 1 to 5 compared to the amino acid sequence of amino acid residues 1 to 127 of SEQ ID NO: 2. The pharmaceutical composition according to claim 1.
  3. The pharmaceutical composition according to claim 1, wherein the first amino acid sequence segment is item A-ii and the second amino acid sequence segment is item D-ii; or, respectively, the first amino acid sequence segment is item B-ii and the second amino acid sequence segment is item E-ii; or, respectively, the first amino acid sequence segment is item C-ii and the second amino acid sequence segment is item F-ii.
  4. A pharmaceutical composition for use in a method of treating a target infection caused by Klebsiella, comprising a protein having cytotoxic activity against Klebsiella, The aforementioned protein, (a) (a-ii) Amino acid sequence of Sequence ID No. 2 (KvarM); Alternatively, (b) (b-ii) an amino acid sequence having at least 90% sequence identity with the amino acid sequence of Sequence ID No. 2; Alternatively, (c) (c-ii) an amino acid sequence having 1 to 30 amino acid substitutions, additions, insertions and/or deletions compared to the amino acid sequence of Sequence ID No. 2, or an amino acid sequence consisting of such, Pharmaceutical composition.
  5. In item (b), the sequence identity is at least 95%; and/or in item (c), the number of amino acid substitutions, additions, insertions and/or deletions is 1 to 20 compared to the amino acid sequence of SEQ ID NO: 2. The pharmaceutical composition according to claim 4.
  6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the protein has cytotoxic activity against Klebsiella pneumoniae, Klebsiella oxytoka, Klebsiella granulomatis, Klebsiella quasipneumoniae, Klebsiella aerogenes, and/or Klebsiella vallicola.
  7. The protein comprises the amino acid sequence of SEQ ID NO: 22, where each X represents one of the 20 standard amino acid residues or the absence of an amino acid residue; or the protein comprises the amino acid sequence of SEQ ID NO: 2. A pharmaceutical composition according to any one of claims 1 to 6.
  8. A pharmaceutical composition according to any one of claims 1 to 7, comprising one or more proteins as defined in any one of claims 1 to 7, and a carrier.
  9. The pharmaceutical composition according to claim 8, wherein the plant material is a plant material or an extract thereof, and the plant material may be a plant-derived material expressing one or more of the aforementioned proteins.
  10. The pharmaceutical composition according to claim 8 or 9, wherein the one or more proteins are formulated for oral delivery to the small or large intestine.
  11. An oral formulation comprising a protein as defined in any one of claims 1 to 7, or a pharmaceutical composition as described in any one of claims 8 to 10, which can protect the protein from gastric conditions and release the protein in the intestine, for use in a method of treating a target infection caused by Klebsiella.
  12. A pharmaceutical composition or formulation according to any one of claims 1 to 11, for use in a method for treating a target infection caused by Klebsiella, wherein the Klebsiella is antibiotic-resistant.
  13. A method for preventing or reducing contamination of an inanimate object by one or more Klebsiella species, comprising the step of contacting the inanimate object with a protein having cytotoxic activity against Klebsiella or a composition containing such protein, wherein the protein is defined in any one of claims 1 to 7.

Description

This invention provides proteins having cytotoxic activity against Klebsiella. The invention also provides compositions, including pharmaceutical compositions containing one or more of the aforementioned proteins. Furthermore, the invention provides proteins having cytotoxic activity against Klebsiella, and compositions containing the protein, for therapeutic use. It also provides proteins or compositions containing the protein, for use in methods of treating infections of a target caused by Klebsiella. Furthermore, it provides oral enteric-coated formulations for delivering the protein or composition to the small or large intestine. Furthermore, it provides pulmonary formulations for delivering the protein or composition to the lungs. It also provides methods for preventing or reducing infection or contamination of objects by Klebsiella, methods for treating Klebsiella infections in a target or patient requiring such treatment, and methods for producing compositions containing the protein. The invention also provides methods for producing compositions containing the protein of the invention. The invention further provides nucleic acid molecules encoding proteins having cytotoxic activity against Klebsiella, plants, plant tissues or plant cells containing the protein, and plants, plant tissues or plants containing the nucleic acid molecule. Klebsiella are non-motile, rod-shaped, Gram-negative bacteria encased in a capsular polysaccharide that confers resistance to many host defense mechanisms. Klebsiella are opportunistic pathogens found in the environment and on the mucous membranes of mammals. Three species of the genus Klebsiella are commonly associated with human diseases: K. pneumoniae, K. oxytoca, and K. granulomatis. Recently, two more Klebsiella species, K. variicola and K. quasipneumoniae, have also been found to cause potentially fatal infections (Long et al. 2017). The primary sources of infection with the pathogen are the patient's gastrointestinal tract and the hands of hospital staff. Outside of hospitals, Klebsiella infection typically occurs in the lungs. The disease typically affects middle-aged and older men with debilitating conditions such as alcoholism, diabetes, or chronic bronchopulmonary spirochetosis (Chan et al., 2009). This patient population is thought to have impaired host defense mechanisms in the respiratory tract. The organism enters the body after the host inhales colonizing oropharyngeal microorganisms into the lower respiratory tract (Hirsche et al., 2005). In recent years, Klebsiella has become a serious pathogen in hospital-acquired infections. Common sites of hospital-acquired infections include the urinary tract, lower respiratory tract, bile duct, and surgical wound sites. The range of clinical syndromes includes pneumonia, bacteremia, thrombophlebitis, urinary tract infections (UTIs), cholecystitis, diarrhea, upper respiratory tract infections, wound infections, osteomyelitis, and meningitis (Miftode et al., 2008). The presence of invasive devices, contamination of respiratory support devices, use of urinary catheters, and use of antibiotics are factors that increase the likelihood of hospital-acquired infections by Klebsiella species (Weisenberg et al., 2009). K. pneumoniae is one of the six pathogens that make up hospital-acquired ESKAPE infections (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter) that rapidly develop antibiotic resistance. In 2016, a highly virulent, carbapenem-resistant K. pneumoniae was reported in China. An outbreak of hospital-acquired pneumonia has been reported, resulting in the deaths of five surgical patients due to infection with the Pneumoniae pneumoniae (CRPK) ST11 strain (Gu et al, 2017). This strain is highly virulent and highly resistant, and can therefore be referred to as a "superbug." The ST11 CR HvKP strain infects relatively healthy populations with normal immunity. These are mucoid strains and adhere to all surfaces in the intensive care unit. Colistin, an antibiotic used as a last resort against carbapenem-resistant enterobacteria, is largely ineffective against this strain. For the time being, ceftazidime/avibactam can be used to treat these infections, but resistance to these antibiotics may soon be acquired. The increasing drug resistance of pathogens observed to date is an international challenge, and the development of a new generation of antimicrobial agents is urgently needed. Bacteria produce toxic proteins called bacteriocins when competing for ecological niches with each other. Bacteriocins typically kill only closely related bacteria belonging to the same species or genus. Their mechanisms of action are diverse, including pore formation, inhibition of DNase and RNase activity, and inhibition of protein synthesis or DNA replication. Bacteriocins produced by Gram-positive bacteria are usually called bacteriocins of a predetermined cl