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EP-4308715-B1 - HYBRID MOLECULE COMPRISING AN ANTIBODY FC FRAGMENT AND AT LEAST ONE FIBRIN-DERIVED CITRULLINATED PEPTIDE, AND USES THEREOF

EP4308715B1EP 4308715 B1EP4308715 B1EP 4308715B1EP-4308715-B1

Inventors

  • COMBES, Eve
  • ROBERT, BRUNO
  • MARTINEAU, PIERRE
  • LOUIS-PLENCE, Pascale
  • VILLALBA, Martin
  • JORGENSEN, CHRISTIAN
  • TROEGELER, Anthony
  • CLAVEL, CYRIL
  • SERRE, Guy, Bruno, René

Dates

Publication Date
20260513
Application Date
20220318

Claims (11)

  1. A hybrid molecule comprising at least one antibody Fc fragment covalently linked to at least one fibrin-derived peptide having at least one citrullyl residue, at least one spacer being optionally present between said Fc fragment and said peptide, said peptide being recognized 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 the preceding claim, said vertebrate fibrin being a mammalian fibrin, more particularly a human fibrin.
  3. A hybrid molecule according to any of the preceding claims, wherein said spacer is a polymer containing one or more repeating units containing the ether group, said spacer preferably being polyethylene glycol of formula PEGn, wherein n represents an integer between 1 and 100, preferably between 1 and 10 and in particular 1, 2, 3, 4 or 8.
  4. 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) wherein 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) wherein 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) wherein X 1 and X 2 each represents 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) wherein 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) wherein 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 the peptides a) to e) above.
  5. A hybrid molecule according to any one of claims 1-3, 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.
  6. A hybrid molecule according to any one of the preceding claims, wherein the said Fc fragment is a human Fc fragment, in particular of IgG, more particularly of IgG1.
  7. A hybrid molecule according to any of the preceding claims, wherein said Fc fragment is wild-type or mutated, said mutated Fc fragment preferably comprising at least the following mutations: - L234A and L235A, or - L234A, L235A and P329G, or - G236A, S239D and I332E, or - G236A, S239D, A330L and I332E, or - S239D, H268F, S324T and I332E, the numbering being indicated in the sequence of a human IgG1 according to the EU index.
  8. A hybrid molecule according to any one of the preceding claims, wherein: - the Fc fragment is coupled to at least one azide and said peptide is coupled to an alkyne, such as a cyclooctyne, and in particular DBCO, or - the Fc fragment is coupled to at least one alkyne, such as a cyclooctyne, and in particular DBCO, and said peptide is coupled to an azide, or - the Fc fragment is coupled to at least one azide and said peptide is bound to a spacer, itself coupled to an alkyne, such as a cyclooctyne, and in particular DBCO, or - the Fc fragment is coupled to at least one alkyne, such as a cyclooctyne, and in particular DBCO, and said peptide is bound to a spacer, itself coupled to an azide, or - the Fc fragment is bound to at least one spacer, itself coupled to an azide, and said peptide is coupled to an alkyne, such as a cyclooctyne, and in particular DBCO, or - the Fc fragment is bound to at least one spacer, itself coupled to an alkyne, such as a cyclooctyne, and in particular DBCO, and said peptide is coupled to an azide, or - the Fc fragment is bound to at least one spacer, itself coupled to an azide, and said peptide is bound to a spacer, itself coupled to an alkyne, such as a cyclooctyne, and in particular DBCO, or - the Fc fragment is bound to at least one spacer, itself coupled to an alkyne, such as a cyclooctyne, and in particular DBCO, and said peptide is bound to a spacer, itself coupled to an azide, the covalent bond between said Fc fragment and said peptide, optionally in the presence of one or more spacers, being created between the azide and the alkyne.
  9. A hybrid molecule according to any one of the preceding claims, said molecule being represented by: - a wild-type Fc fragment that is bound to at least one PEGn spacer that is itself coupled to an azide covalently bound to a cyclooctyne that is coupled to a peptide represented by SEQ ID NO: 19 or SEQ ID NO: 18, - an Fc fragment comprising the S239D, H268F, S324T and I332E mutations that is bound to at least one PEGn spacer that is itself coupled to an azide covalently bound to a cyclooctyne that is coupled to a peptide represented by SEQ ID NO: 19 or SEQ ID NO: 18, - an Fc fragment comprising the L234A, L235A and P329G mutations that is bound to at least one PEGn spacer that is itself coupled to an azide covalently bound to a cyclooctyne that is coupled to a peptide represented by SEQ ID NO: 19 or SEQ ID NO: 18, - a wild-type Fc fragment that is bound to a PEGn spacer that is itself coupled to at least one azide covalently bound to a cyclooctyne coupled to a PEGn spacer that is itself bound to a peptide represented by SEQ ID NO: 19 or SEQ ID NO: 18, - an Fc fragment comprising the S239D, H268F, S324T and I332E mutations that is bound to at least one PEGn spacer that is itself coupled to an azide covalently bound to a cyclooctyne coupled to a PEGn spacer that is 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 or 4.
  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 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.

Description

The present invention relates to a hybrid molecule comprising at least one Fc fragment of antibody covalently linked to at least one fibrin-derived peptide comprising one or more citrullyl residue(s), 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, lymphocytes expressing them on their membranes, 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 the arginyl residues of the polypeptide chains, 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—other autoantibodies 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 the case of rheumatoid arthritis, clones of B lymphocytes express on their membrane immunoglobulins carrying the ACPA specificity (these are ACPA-positive B lymphocytes), while the plasma cells resulting from the differentiation of ACPA-positive B lymphocyte clones (these are ACPA-positive plasma cells) secrete these same ACPAs in abundance into their microenvironment. The present invention is based on the Inventors' findings that it is possible to target and eliminate B lymphocyte clones expressing ACPA on their surface using a hybrid molecule comprising (i) at least one fibrin-derived peptide having at least one citrullyl residue and (ii) an Fc fragment of human immunoglobulin. These hybrid molecules will specifically target ACPA-positive B lymphocytes (using the fibrin-derived peptide having at least one citrullyl residue, which is recognized by said ACPA), which will then be eliminated, after binding of the Fc fragment to Fc receptors, by macrophages (via phagocytosis) and/or NK cells (via antibody-dependent cell-mediated cytotoxicity - ADCC), and/or by activation of the complement cascade. By targeting B lymphocyte clones expressing ACPAs and cells that differentiate into plasma cells that themselves secrete ACPAs, the hybrid molecules of the invention aim to eliminate ACPAs from the patients' bodies. Description of the invention Hybrid molecule according to the invention In a first aspect, the invention relates to a hybrid molecule comprising at least one Fc fragment of an antibody covalently linked to at least one fibrin-derived peptide having at least one citrullyl residue, at least one spacer being optionally present between said Fc fragment and said peptide, said peptide being recognized by anti-citrullinated protein autoantibodies and being derived from all or part