Search

JP-2026076233-A - Chimeric polypeptide

JP2026076233AJP 2026076233 AJP2026076233 AJP 2026076233AJP-2026076233-A

Abstract

[Problem] To provide a vaccine for use in reducing the incidence and/or severity of periodontal pathogen Porphyromonas gingivalis (P. gingivalis) infection, or for treating P. gingivalis infection and disease in a target population. [Solution] A chimeric or fusion protein for inducing an immune response against P. gingivalis is provided, comprising a protein first polypeptide and a second polypeptide, wherein A) the first polypeptide contains or consists of the amino acid sequence of the active site of Arg- or Lys-gingipain of P. gingivalis, or a sequence that is at least 80% identical thereto, and B) the second polypeptide contains or consists of the amino acid sequence of the adhesion factor domain of Arg- or Lys-gingipain of P. gingivalis. [Selection Diagram] None

Inventors

  • レイノルズ，エリック チャールズ
  • スラケスキー，ナダ
  • オブライエン－シンプソン，ニール マーティン

Assignees

  • デントリック ピーティーワイ エルティーディー

Dates

Publication Date
20260511
Application Date
20260120
Priority Date
20220120

Claims (20)

  1. A chimeric or fusion protein for inducing an immune response against P. gingivalis, wherein the protein comprises a first polypeptide and a second polypeptide. A) The first polypeptide contains, or comprises, the amino acid sequence of the active site of Arg- or Lys-gingipain of P. gingivalis, or a sequence that is at least 80% identical thereto. B) The second polypeptide contains or consists of the amino acid sequence of the adhesion factor domain of Arg- or Lys-gingipain of P. gingivalis. The second polypeptide comprises a sequence of one or more adhesion factor-binding motifs (ABMs), The second polypeptide comprises a portion or all of the sequence of the cleaved adhesion factor domain (CAD), and the second polypeptide comprises a) an amino acid sequence substantially corresponding to the full length of the DUF2436 domain of Arg- or Lys-gingipain, or a sequence that is at least 80% identical thereto, b) Compared to naturally occurring Arg- or Lys-gingipain sequences in the corresponding region, it contains one or more cysteine amino acid substitutions in the adhesion factor domain, and c) i) Substitution of proline and/or asparagine residues in sequence PxxN at positions corresponding to or equivalent to residues 6-9 of sequence SEQ ID NO: 14 or 19 (ABM1), ii) Substitution of motif NxFA with SxYQ in sequences corresponding to or equivalent to residues 2-5 of sequence number 14 or 19 (ABM1), iii) The chimeric or fusion protein comprising one or more amino acid motif substitutions selected from: a second tyrosine residue at a position corresponding to or equivalent to the residue at position 5 of SEQ ID NO: 15 or 20 (ABM2), and a substitution of a tryptophan residue at a position corresponding to or equivalent to the residue at position 23 of SEQ ID NO: 14 or 19 (ABM1) with an alanine residue.
  2. The chimeric or fusion protein according to claim 1, wherein one or more ABMs include the sequence described in SEQ ID NO: 15 or 20, SEQ ID NO: 14 or 19, and/or SEQ ID NO: 17 or 21, or a sequence that is at least 80% identical to those sequences.
  3. The chimeric or fusion protein according to claim 1 or 2, wherein one or more ABMs contain the sequence described in SEQ ID NO: 16 or SEQ ID NO: 18 or 22, or a sequence that is at least 80% identical to those sequences.
  4. The chimeric or fusion protein according to any one of claims 1 to 3, wherein the second polypeptide comprises a portion or all of the sequence of CAD having at least 80% the amino acid sequence described in SEQ ID NO: 12 or 13, or a sequence identical thereto to those described therein.
  5. The chimeric or fusion protein according to any one of claims 1 to 4, wherein the amino acid sequence substantially corresponding to the full length of the DUF2436 domain of Arg- or Lys-gingipain is a sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the length of the DUF2436 domain of Arg- or Lys-gingipain.
  6. The chimeric or fusion protein according to claim 5, wherein the amino acid sequence of the DUF2436 domain of Arg- or Lys-gingipain is the sequence described in SEQ ID NO: 23, or at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto.
  7. The chimeric or fusion protein according to any one of claims 1 to 6, wherein the second polypeptide comprises the sequence described in SEQ ID NO: 33, or at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that sequence.
  8. The chimeric or fusion protein according to any one of the prior claims, wherein the second polypeptide does not involve substitution of cysteine residues in the DUF2436 domain.
  9. The chimeric or fusion protein according to any one of claims 1 to 7, wherein the second polypeptide comprises the substitution of the cysteine residue in the DUF2436 domain.
  10. The chimeric or fusion protein according to claim 9, wherein the substitution of the cysteine residue is a substitution of a serine or valine residue.
  11. The chimeric or fusion protein according to claim 10, wherein the substitution of the cysteine residue is a substitution of a serine residue.
  12. The chimeric or fusion protein according to any one of claims 1 to 11, wherein the second polypeptide comprises one or more substitutions of the cysteine residues in the ABM domain.
  13. The chimeric or fusion protein according to claim 12, wherein the substitution of one or more cysteine residues is a substitution of a serine residue or a valine residue.
  14. The chimeric or fusion protein according to claim 13, wherein the substitution of one or more cysteine residues is a substitution of serine residues.
  15. The chimeric or fusion protein according to any one of claims 12 to 14, wherein the sequence of one or more ABM domains is described in SEQ ID NO: 18 or 22, and the one or more cysteine residues correspond to the residues at positions 36 and 50 of SEQ ID NO: 18 or 22.
  16. The chimeric or fusion protein according to any one of the prior claims, wherein the second polypeptide comprises substitutions of proline and/or sparagine residues in the sequence PxxN of the ABM1 of the P. gingivalis adhesion factor domain at positions corresponding to or equivalent to residues 6-9 of the sequence of SEQ ID NO: 14 or 19.
  17. The chimeric or fusion protein according to claim 16, wherein the second polypeptide comprises the sequence AxxN, AxxA, or AxxP in ABM1 at positions corresponding to or equivalent to residues 6-9 of the sequence of SEQ ID NO: 14 or 19.
  18. The chimeric or fusion protein according to any one of the prior claims, wherein the second polypeptide comprises or consists of the amino acid sequence described in any one of SEQ ID NOs: 33 to 48.
  19. The chimeric or fusion protein according to claim 18, wherein the second polypeptide comprises or consists of the amino acid sequence described in SEQ ID NO: 39 or 40.
  20. The chimeric or fusion protein according to claim 17, wherein the second polypeptide comprises or consists of the amino acid sequence described in SEQ ID NO: 47 or 48.

Description

This invention relates to chimeric polypeptides useful for inducing an immune response against P. gingivalis, compositions comprising the same, and their use for the prevention and treatment of P. gingivalis-related conditions and diseases. Related Application This application claims priority from Australian Provisional Application No. 2022/900103, the entirety of which is incorporated herein by reference. If plaque accumulates around the teeth at the gum line, it causes inflammation of the gums (gingivitis). Chronic gingivitis can lead to the development of the periodontal pathogen *Porphyromonas gingivalis* (P. gingivalis) at the base of the periodontal pocket, potentially resulting in chronic infection and severe disease. This severe form of periodontal disease, called periodontitis, can lead to tooth loss, as the immune system's response to eliminate infection is ineffective. Chronic periodontitis is an inflammatory disease of the supporting tissues of teeth, leading to alveolar bone resorption and ultimately tooth loss. This disease is a major public health issue in all societies, affecting up to 30% of the adult population, with an estimated 12–15% of adults suffering from a severe form. One in three adults has moderate to severe periodontitis. Epidemiological studies have shown that periodontitis is associated with an increased risk of cardiovascular disease, certain cancers, premature birth, rheumatoid arthritis, and inflammatory diseases, including dementia. More recent studies have linked chronic infection with P. gingivalis to dementia and rheumatoid arthritis. For example, one study showed that 96% of Alzheimer's disease (AD) brain samples had P. gingivalis. Another study showed that chronic oral infection with P. gingivalis in mice resulted in AD-associated brain plaques in humans, and that P. gingivalis proteases can cleave amyloid precursors and tau proteins to form AD-associated plaques and tangles. Several pathogenic factors contributing to the pathogenicity of P. gingivalis have been reported, including LPS, fimbriae, hemagglutinins, hemolysins, and extracellular hydrolases (particularly Arg-X and Lys-X specific proteinases), also known as "P. gingivalis gingipain." The magnitude of this public health problem suggests that a vaccine and means of providing it that offer a robust protective response against P. gingivalis infection are necessary. One problem was the lack of a clear method for obtaining a robust protective response against P. gingivalis infection, which has an excessive number of pathogenic factors to select. Currently, there are no commercially approved vaccines for use in preventing or reducing the incidence and/or severity of P. gingivalis infection, or for treating P. gingivalis infection and disease in the target population. Therefore, alternative and/or improved approaches to the design and manufacture of P. gingivalis vaccines, and alternative and/or improved vaccines produced from P. gingivalis, are needed. Any reference to prior art in this specification does not constitute an admission or suggestion that such prior art forms part of the common general knowledge in any jurisdiction, or that such prior art can be reasonably expected to be understood, considered relevant, and/or combined with other prior art by those skilled in the art. The present invention provides a chimeric or fusion protein for inducing an immune response against P. gingivalis, wherein the protein comprises a first polypeptide and a second polypeptide. A) The first polypeptide contains, or consists of, the amino acid sequence of the active site of Arg- or Lys-gingipain of P. gingivalis, or a sequence that is at least 80% identical thereto. B) The second polypeptide contains or consists of the amino acid sequence of the adhesion factor domain of Arg- or Lys-gingipain of P. gingivalis. The second polypeptide comprises a sequence of one or more adhesion factor-binding motifs (ABMs), preferably the ABMs correspond to part or all of the ABMs between the DUF2436 domain and the cleaved adhesion factor domain (CAD) of P. gingivalis gingipain. Preferably, one or more ABMs include the sequence described in SEQ ID NO: 15 or 20 (ABM2) and/or SEQ ID NO: 17 or 21 (ABM3), or a sequence that is at least 80% identical to them. More preferably, one or more ABMs include the sequence described in SEQ ID NO: 15 or 20 (ABM2), SEQ ID NO: 14 or 19 (ABM1), and/or SEQ ID NO: 17 or 21 (ABM3), or a sequence that is at least 80% identical to them. Most preferably, one or more ABMs include the sequence described in SEQ ID NO: 16, or SEQ ID NO: 18 or 22, or SEQ ID NO: 27, 63, or 64, or a sequence that is at least 80% identical to those sequences. The second polypeptide comprises a portion or all of a cleaved adhesion factor domain (CAD), preferably the amino acid sequence described in SEQ ID NO: 12 or 13, or a portion or all of a sequence of CAD having at least 80% identical to those sequences. The second polypeptide is a) compr