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EP-4308716-B1 - HYBRID MOLECULE COMPRISING A FIBRIN-DERIVED CITRULLINATED PEPTIDE AND AN ANTIBODY OR ANTIBODY FRAGMENT WHICH BINDS TO CD38 AND/OR CD138, AND USES THEREOF

EP4308716B1EP 4308716 B1EP4308716 B1EP 4308716B1EP-4308716-B1

Inventors

  • VACHIN, Pauline
  • LOUIS-PLENCE, Pascale
  • JORGENSEN, CHRISTIAN
  • ESQUERRE, Camille
  • CLAVEL, CYRIL
  • SERRE, Guy, Bruno, René
  • COMBES, Eve
  • ROBERT, BRUNO
  • MARTINEAU, PIERRE

Dates

Publication Date
20260513
Application Date
20220318

Claims (11)

  1. A hybrid molecule comprising at least one fibrin-derived peptide having at least one citrullyl residue, said peptide being covalently bound to at least one antibody or to at least one antibody fragment, said antibody or fragment being capable of binding to CD38 and/or CD138, one or more spacers being optionally present between said peptide and said antibody or said fragment, said peptide being recognised by the anti-citrullinated protein autoantibodies and being derived from all or part of the sequence of the α or β chain of a vertebrate fibrin by substitution of at least one arginyl residue by a citrullyl residue.
  2. A hybrid molecule according to claim 1, wherein said peptide is covalently bound to a spacer, said spacer being itself covalently bound to said antibody or antibody fragment.
  3. A hybrid molecule according to either one of the preceding claims, said vertebrate fibrin being a mammalian fibrin, preferably a human fibrin.
  4. A hybrid molecule according to any one of the preceding claims, wherein said spacer is a polymer containing one or more repeat units containing the ether group, said spacer preferably being polyethylene glycol of formula PEGn, in which n represents an integer between 1 and 100, preferably between 1 and 10 and in particular 1, 2, 3, 4 or 8.
  5. A hybrid molecule according to any one of the preceding claims, wherein said peptide comprises at least one citrullyl residue, and is selected from the group consisting of: a) a peptide defined by the sequence X 1 PAPPPISGGGYX 2 AX 3 (SEQ ID NO: 1) in which X 1 , X 2 , and X 3 each represent a citrullyl residue or an arginyl residue, and at least one of the X 1 or X 2 or X 3 residues is a citrullyl residue; b) a peptide defined by the sequence GPX 1 VVEX 2 HQSACKDS (SEQ ID NO: 2) in which X 1 and X 2 each represent a citrullyl residue or an arginyl residue, and at least one of the X 1 or X 2 residues is a citrullyl residue; c) a peptide defined by the sequence SGIGTLDGFX 1 HX 2 HPD (SEQ ID NO:3) in which X 1 and X 2 each represent a citrullyl residue or an arginyl residue, and at least one of the X 1 or X 2 residues is a citrullyl residue; d) a peptide defined by the sequence VDIDIKIX 1 SCX 2 GSCS (SEQ ID NO:4) in which X 1 and X 2 each represent a citrullyl residue or an arginyl residue, and at least one of the X 1 or X 2 residues is a citrullyl residue; e) a peptide defined by the sequence X 1 GHAKSX 2 PVX 3 GIHTS (SEQ ID NO: 12) in which X 1 , X 2 and X 3 each represent a citrullyl residue or an arginyl residue, and at least one of the X 1 or X 2 or X 3 residues is a citrullyl residue; f) a peptide comprising at least 5 consecutive amino acids, including at least one citrullyl residue, from one of peptides a) to e) above.
  6. A hybrid molecule according to any one of claims 1-4, wherein said peptide is selected from the group consisting of: SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, more particularly selected from the group consisting of: SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 14, SEQ ID NO: 18 and SEQ ID NO: 19, more particularly SEQ ID NO: 18 and SEQ ID NO: 19.
  7. A hybrid molecule according to any one of the preceding claims, wherein the fragment is selected from fragments, scFc, Fv, Fab or F(ab') 2 .
  8. A hybrid molecule according to any one of the preceding claims, wherein said antibody or F(ab') 2 fragment is a bispecific antibody or F(ab') 2 fragment, directed against: - CD38 and another plasma cell target, or - CD138 and another plasma cell target, or - CD38 and CD138.
  9. A hybrid molecule according to any one of the preceding claims, said molecule being represented by: - an antibody fragment capable of binding to CD38 bound to a PEGn spacer, itself coupled to an azide covalently bound to a cyclooctyne coupled to a peptide represented by SEQ ID NO: 19, or SEQ ID NO: 18, - an antibody fragment capable of binding to CD38 bound to a PEGn spacer, itself coupled to an azide covalently bound to a cyclooctyne coupled to a PEGn spacer, itself bound to a peptide represented by SEQ ID NO:19 or SEQ ID NO: 18, n preferably representing an integer between 1 and 10, more particularly 1, 2, 3, 4 or 8.
  10. A hybrid molecule according to any one of the preceding claims for use as a medicinal product, in particular for use in the treatment of autoimmune diseases associated with the production of anti-citrullinated protein autoantibodies, in particular Gougerot-Sjögren syndrome and rheumatoid arthritis.
  11. A hybrid molecule according to any one of the preceding claims, for use as a medicinal product, in combination with a second hybrid molecule, said second molecule comprising at least one Fc antibody fragment covalently bound to at least one fibrin-derived peptide carrier of a citrullyl residue, a spacer being optionally present between said Fc fragment and said peptide.

Description

The present invention relates to a hybrid molecule comprising a citrullinated fibrin-derived peptide and an antibody or antibody fragment binding to CD38 and/or CD138, the uses of such a hybrid molecule, and its production process. Context of the invention Rheumatoid arthritis is the most common inflammatory rheumatic disease, and also the most common autoimmune disease. The disease is characterized by chronic inflammation of the synovial joints, leading to irreversible joint destruction. The presence of class G autoantibodies directed against citrullinated proteins, known as anti-citrullinated protein autoantibodies (ACPA), is highly specific for rheumatoid arthritis. Patient sera containing ACPA, the lymphocytes that express them on their membrane, and the patients themselves are said to be 'ACPA-positive'. Several studies have demonstrated that these ACPA are central to the autoimmune reactions characteristic of rheumatoid arthritis and thus represent a promising therapeutic target. The antigenic targets of ACPAs have been characterized. They are specifically directed against deiminated or citrullinated forms of the α and β polypeptide chains of fibrin, a protein abundant in inflammatory synovial tissue. This citrullination corresponds to the enzymatic deimination of arginyl residues of a protein, under the action of peptidyl-arginine deiminases (PADs). More specifically, the immunodominant epitopes recognized by ACPA on the α and β polypeptide chains of fibrin have been characterized and published, notably in applications PCT/FR00/01857 Or PCT/FR2007/000758 The five citrullinated peptides carrying immunodominant epitopes are specifically named α36-50Cit (as represented in SEQ ID NO: 5), α171-185Cit (as represented in SEQ ID NO: 6), α501-515Cit (as represented in SEQ ID NO: 14), α621-635Cit (as represented in SEQ ID NO: 18), and β60-74Cit (as represented in SEQ ID NO: 19). Sera from ACPA-positive patients recognize one or more of these five peptides. Secreted into the rheumatoid synovial tissue by local plasma cells, ACPAs are present in high concentrations, close to their main target, citrullinated fibrin, which is also abundant in the interstitial deposits. The binding of ACPAs to these deposits, and thus the formation of immobilized immune complexes, which in turn bind rheumatoid factors—another autoantibody associated with rheumatoid arthritis and also secreted by local plasma cells—triggers a cascade of pro-inflammatory events. Stimulation of macrophage cells by these immune macro-complexes, primarily via their membrane Fcgamma receptors, leads them to secrete cytokines. pro-inflammatory and in particular TNF-alpha which has been identified as the main cytokine responsible for rheumatoid inflammation. Currently, there is no cure for rheumatoid arthritis. Treatments are only aimed at managing flare-ups and/or preventing their occurrence. One object of the present invention is thus to provide a treatment for rheumatoid arthritis. ACPAs are oligoclonal and therefore secreted by only a few plasma cell clones, themselves resulting from the differentiation of a few B lymphocyte clones. In cases of rheumatoid arthritis, clones of B lymphocytes express on their membrane immunoglobulins carrying the ACPA specificity (these are ACPA-positive B lymphocytes), while plasma cells resulting from the differentiation of ACPA-positive B lymphocyte clones (these are ACPA-positive plasma cells) secrete ACPA in abundance into their microenvironment. The present invention is based on the Inventors' results showing that it is possible to target and eliminate plasma cells that abundantly secrete ACPAs in their microenvironment using a hybrid molecule comprising (i) a fibrin-derived peptide having at least one citrullyl residue and (ii) an antibody or antibody fragment binding to CD38 and/or CD138. These hybrid molecules will target the plasma cells (using the antibody or antibody fragment binding to CD38 and/or CD138, which are molecular markers expressed on the surface of said plasma cells) and, thanks to the fibrin-derived peptide having at least one citrullyl residue, the hybrid molecule will be specifically recognized by the ACPAs (the peptide being the target of these ACPAs). After bridging (ACPA binding to the hybrid molecule, itself bound to plasma cells), the plasma cells will be lysed by apoptosis, by phagocytosis in the presence of macrophages, by ADCC (antibody-dependent cell-mediated cytotoxicity) in the presence of NK cells, or by complement activation. By targeting plasma cells that secrete ACPA, the hybrid molecules of the invention aim to eliminate ACPA from the patients' bodies and the source of these ACPA in order to induce remission of the disease. The targeting of plasma cells and the antibodies released by said plasma cells using hybrids has already been described, for example in the application EP2892926 However, no specific construct or outcome related to rheumatoid arthritis is described. Descripti