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EP-4735478-A1 - NEUTRALIZING HUMAN MONOCLONAL ANTIBODIES AGAINST P. AERUGINOSA

EP4735478A1EP 4735478 A1EP4735478 A1EP 4735478A1EP-4735478-A1

Abstract

The present disclosure relates to antibodies or antigen-binding fragments thereof against Pseudomonas aeruginosa, pharmaceutical compositions comprising such antibodies or antigen-binding fragments thereof, kits comprising such antibodies or antigen-binding fragments thereof, and the antibodies or antigen-binding fragments thereof, the pharmaceutical compositions and the kits for use as a medicament, and in the treatment or prevention of a disease caused by Pseudomonas aeruginosa. The present disclosure further relates to methods of treating, preventing or reducing the severity of an infection with Pseudomonas aeruginosa, and to nucleic acids encoding such antibodies or antigen-binding fragments thereof, expression vectors comprising such nucleic acids, host cells comprising such nucleic acids or expression vectors, and methods for the production of such antibodies or antigen-binding fragments thereof.

Inventors

  • KLEIN, FLORIAN
  • RYBNIKER, Jan
  • SIMONIS, ALEXANDER
  • KREER, Christoph

Assignees

  • Universität zu Köln

Dates

Publication Date
20260506
Application Date
20240627

Claims (15)

  1. 1. Antibody or antigen-binding fragment thereof directed against Pseudomonas aeruginosa, wherein the antibody or antigen-binding fragment thereof comprises the combination of the heavy chain CDR1 to CDR3 and the light chain CDR1 to CDR3 amino acid sequence of one antibody selected from the group comprising: 30-D9 (having a CDR-H1 amino acid sequence of SEQ ID No. 21 , a CDR-H2 amino acid sequence of SEQ ID No. 22, a CDR-H3 amino acid sequence of SEQ ID No. 23, a CDR-L1 amino acid sequence of SEQ ID No. 24, a CDR-L2 amino acid sequence of SEQ ID No. 25, a CDR-L3 amino acid sequence of SEQ ID No. 26), 30-B8 (having a CDR-H1 amino acid sequence of SEQ ID No. 27, a CDR-H2 amino acid sequence of SEQ ID No. 28, a CDR-H3 amino acid sequence of SEQ ID No. 29, a CDR-L1 amino acid sequence of SEQ ID No. 30, a CDR-L2 amino acid sequence of SEQ ID No. 31 , a CDR-L3 amino acid sequence of SEQ ID No. 32), 30-D7 (having a CDR-H1 amino acid sequence of SEQ ID No. 33, a CDR-H2 amino acid sequence of SEQ ID No. 34, a CDR-H3 amino acid sequence of SEQ ID No. 35, a CDR-L1 amino acid sequence of SEQ ID No. 36, a CDR-L2 amino acid sequence of SEQ ID No. 37, a CDR-L3 amino acid sequence of SEQ ID No. 38), 11-A6 (having a CDR-H1 amino acid sequence of SEQ ID No. 39, a CDR-H2 amino acid sequence of SEQ ID No. 40, a CDR-H3 amino acid sequence of SEQ ID No. 41 , a CDR-L1 amino acid sequence of SEQ ID No. 42, a CDR-L2 amino acid sequence of SEQ ID No. 43, a CDR-L3 amino acid sequence of SEQ ID No. 44), 23-A9 (having a CDR-H1 amino acid sequence of SEQ ID No. 45, a CDR-H2 amino acid sequence of SEQ ID No. 46, a CDR-H3 amino acid sequence of SEQ ID No. 47, a CDR-L1 amino acid sequence of SEQ ID No. 48, a CDR-L2 amino acid sequence of SEQ ID No. 49, a CDR-L3 amino acid sequence of SEQ ID No. 50), 11-C10 (having a CDR-H1 amino acid sequence of SEQ ID No. 51 , a CDR-H2 amino acid sequence of SEQ ID No. 52, a CDR-H3 amino acid sequence of SEQ ID No. 53, a CDR-L1 amino acid sequence of SEQ ID No. 54, a CDR-L2 amino acid sequence of SEQ ID No. 55, a CDR-L3 amino acid sequence of SEQ ID No. 56), 23-F9 (having a CDR-H1 amino acid sequence of SEQ ID No. 57, a CDR-H2 amino acid sequence of SEQ ID No. 58, a CDR-H3 amino acid sequence of SEQ ID No. 59, a CDR-L1 amino acid sequence of SEQ ID No. 60, a CDR-L2 amino acid sequence of SEQ ID No. 61 , a CDR-L3 amino acid sequence of SEQ ID No. 62), 11 -C4 (having a CDR-H1 amino acid sequence of SEQ ID No. 63, a CDR-H2 amino acid sequence of SEQ ID No. 64, a CDR-H3 amino acid sequence of SEQ ID No. 65, a CDR-L1 amino acid sequence of SEQ ID No. 66, a CDR-L2 amino acid sequence of SEQ ID No. 67, a CDR-L3 amino acid sequence of SEQ ID No. 68), 30-B9 (having a CDR-H1 amino acid sequence of SEQ ID No. 69, a CDR-H2 amino acid sequence of SEQ ID No. 70, a CDR-H3 amino acid sequence of SEQ ID No. 71 , a CDR-L1 amino acid sequence of SEQ ID No. 72, a CDR-L2 amino acid sequence of SEQ ID No. 73, a CDR-L3 amino acid sequence of SEQ ID No. 74), and 30-C9 (having a CDR-H1 amino acid sequence of SEQ ID No. 75, a CDR-H2 amino acid sequence of SEQ ID No. 76, a CDR-H3 amino acid sequence of SEQ ID No. 77, a CDR-L1 amino acid sequence of SEQ ID No. 78, a CDR-L2 amino acid sequence of SEQ ID No. 79, a CDR-L3 amino acid sequence of SEQ ID No. 80).
  2. 2. Antibody or antigen-binding fragment thereof according to claim 1 , wherein the antibody or antigen-binding fragment thereof comprises the combination of the variable region heavy chain amino acid sequence and of the variable region light chain amino acid sequence of one antibody selected from the group comprising: 30-D9 (having the variable region heavy chain amino acid sequence of SEQ ID No. 1 and the variable region light chain amino acid sequence of SEQ ID No. 2), 30-B8 (having the variable region heavy chain amino acid sequence of SEQ ID No. 3 and the variable region light chain amino acid sequence of SEQ ID No. 4), 30-D7 (having the variable region heavy chain amino acid sequence of SEQ ID No. 5 and the variable region light chain amino acid sequence of SEQ ID No. 6), 11-A6 (having the variable region heavy chain amino acid sequence of SEQ ID No. 7 and the variable region light chain amino acid sequence of SEQ ID No. 8), 23-A9 (having the variable region heavy chain amino acid sequence of SEQ ID No. 9 and the variable region light chain amino acid sequence of SEQ ID No. 10), 11-C10 (having the variable region heavy chain amino acid sequence of SEQ ID No. 11 and the variable region light chain amino acid sequence of SEQ ID No. 12), 23-F9 (having the variable region heavy chain amino acid sequence of SEQ ID No. 13 and the variable region light chain amino acid sequence of SEQ ID No. 14), 11 -C4 (having the variable region heavy chain amino acid sequence of SEQ ID No. 15 and the variable region light chain amino acid sequence of SEQ ID No. 16), 30-B9 (having the variable region heavy chain amino acid sequence of SEQ ID No. 17 and the variable region light chain amino acid sequence of SEQ ID No. 18), and 30-C9 (having the variable region heavy chain amino acid sequence of SEQ ID No. 19 and the variable region light chain amino acid sequence of SEQ ID No. 20).
  3. 3. Antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the amino acid sequences comprised are of one antibody selected from the group comprising, 30-D9, 30-B8, 30-D7, 11-A6, 23-A9, and 11-C10, preferably of one antibody from the group comprising 30-D9, 30-B8, 30-D7, and 11-A6, more preferably of one antibody from the group comprising 30-D9 and 30-B8, particularly preferably of the antibody 30-B8.
  4. 4. Antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, wherein the antibody or antigen-binding fragment thereof is directed against a protein of the type III secretion system of Pseudomonas aeruginosa, preferably against the protein PcrV of Pseudomonas aeruginosa (UniProt accession number G3XD49).
  5. 5. Antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, wherein the amino acid sequences of the CDRs or of the variable regions comprised therein are from an antibody which is able to inhibit Pseudomonas aeruginosa wild type strain PAO1 in a cytotoxicity assay as described in the description with an IC50 of at most 2 pg/ml, preferably at most 1 pg/ml, more preferably at most 0.3 pg/ml, even more preferably at most 0.2 pg/ml, particularly preferably at most 0.1 pg/ml.
  6. 6. Antibody or antigen-binding fragment thereof according to any one of claims 1 to 5, wherein the amino acid sequences of the CDRs or of the variable regions comprised therein are from an antibody which is able to inhibit Pseudomonas aeruginosa strain PAO1 -induced cell death to obtain cell viability in comparison to uninfected controls in an assay as described in the description with at least 80% viability, preferably at least 84% viability, more preferably at least 90% viability, particularly preferably at least 95% viability.
  7. 7. Antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, wherein the antibody or antigen-binding fragment thereof does not display autoreactivity defined as detectable binding when tested against permeabilized HEp-2 cells using an antinuclear antibody (ANA) testing kit (NOVA-Lite HEp-2 ANA kit; Inova Diagnostics) at concentrations of 100 pg/ml of the antibody or binding fragment thereof.
  8. 8. Pharmaceutical composition comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 7 and at least one pharmaceutically acceptable excipient.
  9. 9. Kit comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 7 and a container.
  10. 10. Antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, pharmaceutical composition according to claim 8, or kit according to claim 9 for use as a medicament.
  11. 11. Antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, pharmaceutical composition according to claim 8, or kit according to claim 9 for use in the treatment or prevention of an infection with Pseudomonas aeruginosa in mammalian subjects, preferably in human subjects.
  12. 12. Nucleic acid encoding an antibody or antigen-binding fragment thereof according to any of claims 1 to 7.
  13. 13. An expression vector comprising the nucleic acid of claim 12 in functional association with an expression control sequence.
  14. 14. Host cell comprising a nucleic acid according to claim 12 or the expression vector according to claim 13.
  15. 5. Method of production of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, comprising (a) cultivating the host cell of claim 14 under conditions allowing expression of the antibody or antigen-binding fragment thereof, and (b) recovering the antibody or antigen-binding fragment thereof.

Description

NEUTRALIZING HUMAN MONOCLONAL ANTIBODIES AGAINST P. AERUGINOSA Technical Field The present invention relates to antibodies or antigen-binding fragments thereof against Pseudomonas aeruginosa, pharmaceutical compositions comprising such antibodies or antigenbinding fragments thereof, kits comprising such antibodies or antigen-binding fragments thereof, and the antibodies or antigen-binding fragments thereof, the pharmaceutical compositions and the kits for use as a medicament, and in the treatment or prevention of a disease caused by Pseudomonas aeruginosa. The present invention further relates to methods of treating, preventing or reducing the severity of an infection with Pseudomonas aeruginosa, and to nucleic acids encoding such antibodies or antigen-binding fragments thereof, expression vectors comprising such nucleic acids, host cells comprising such nucleic acids or expression vectors, and methods for the production of such antibodies or antigen-binding fragments thereof. Technological Background Antimicrobial resistance is an emerging global threat with increasing morbidity and mortality worldwide. This critical situation is aggravated by an innovation and discovery gap leading to a tremendous lack of substances with antibacterial activity. Alternative approaches such as antibody or bacteriophage-based therapies as well as anti-virulence or host directed drugs are required to meet the global needs of therapeutics active against drug-resistant bacteria. In the last two decades, several studies demonstrated the therapeutic potential of neutralizing antibodies against viral infections. Here, broadly neutralizing antibodies (bNAbs) were mainly identified by performing a comprehensive assessment of antigen-reactive B cells derived from infected, convalescent, or vaccinated individuals. However, while numerous antibodies have been developed to target viral pathogens, antibody-mediated treatment approaches against bacterial pathogens were rarely successful and only a limited number of antibodies were identified to potently neutralize bacterial pathogens. Pseudomonas aeruginosa (hereinafter also referred to as P. aeruginosa or PA) is a Gram-negative pathogen which frequently causes severe nosocomial infections including pneumonia and sepsis. Pseudomonas aeruginosa has been classified as a serious threat to the public health by the Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO) due to extensive intrinsic and extrinsic resistance mechanisms. Besides acute infections, Pseudomonas aeruginosa is also capable of causing chronic infections, for instance in patients with structural lung diseases such as chronic obstructive pulmonary disease (COPD) or cystic fibrosis (CF), a monogenetic disease determined by Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) mutations. Decreased mucociliary clearance of the bronchial system and production of a nutrient-rich, hyper-viscous airway mucus in this disease provide ideal growth conditions for opportunistic pathogens such as Pseudomonas aeruginosa. A key virulence factor of Pseudomonas aeruginosa is the type III secretion system (T3SS), a syringelike, multiprotein structure which injects effector toxins such as ExoS and Exoll into the cytosol of host cells leading to cell lysis and tissue damage. The T3SS has been linked to bacterial persistence, higher relapse rates and increased mortality in infected patients. PcrV, a pentameric structural protein forms the T3SS needle-tip complex which is required for appropriate assembly of the PopB/D translocon complex and its insertion into the host cell membrane. As immunogenicity of PcrV has been known for decades, several works have been focusing on antibody-mediated abrogation of PcrV function to inhibit virulence of Pseudomonas aeruginosa, for instance, US 2005/0063985 A1 , US 2013/0108627 A1 , and a study by DiGiandomenico (DiGiandomenico et al., 2014). Safety, efficacy, and pharmacokinetics of the anti-PcrV monoclonal antibody of DiGiandomenico A et al. was evaluated in a clinical trial, where it was found that the anti-PcrV monoclonal antibody was unable to reduce the incidence of nosocomially-acquired pneumonia caused by Pseudomonas aeruginosa in mechanically ventilated patients infected with Pseudomonas aeruginosa. Previous works rely on immunization of mice to generate PcrV-specific antibody sequences, while in-depth investigations of the human B cell response to PcrV and subsequent exploitation of the B cell repertoire for the development of patient-derived highly neutralizing antibodies are lacking so far. As Pseudomonas aeruginosa can reside over years in the airways of people with CF (pwCF), it is hypothesized that the repetitive antigen exposure in these patients fosters a highly affinity matured adaptive immune response, which results in the development of antibodies potently inhibiting virulence of Pseudomonas aeruginosa. Therefore, it is essential to develop fully hu