Antibodies binding to citrullinated histone 2a and/or 4

Abstract

The invention provides antibodies or binding fragments thereof directed against citrulline-containing epitopes. The antibodies or binding fragments thereof of the invention can be used in therapy, for example in the treatment or prevention of Neutrophil Extracellular Trap (NET)-associated pathologies.

1 . An antibody or binding fragment thereof that specifically binds to a citrullinated epitope on deiminated human histone 2A and/or histone 4, wherein the antibody or binding fragment thereof comprises:
 a) CDR1 of VL, wherein the CDR comprises or consists of the amino acid sequence QSL-X 1 -D-X 2 -D-X 3 -KTY, wherein X 1  is V or L, X 2  is T, S, A or N and X 3  is G or A (SEQ ID NO:57), provided that the amino acid sequence is not QSLLDSDGKTY (SEQ ID NO: 36) or QSLVDSDGKTY (SEQ ID NO: 37); and   b) at least one CDR selected from: CDR1 of VH set forth in SEQ ID NO:1, CDR2 of VH set forth in SEQ ID NO:2, CDR3 of VH set forth in SEQ ID NO:3, CDR2 of VL set forth in SEQ ID NO:4, and CDR3 of VL set forth in SEQ ID NO:5.   
     
     
         2 . The antibody or binding fragment thereof of  claim 1 , wherein the antibody or binding fragment thereof comprises:
 a) CDR1 of VL, wherein the CDR comprises or consists of the amino acid sequence QSL-X 1 -D-X 2 -D-X 3 -KTY, wherein X 1  is V or L, X 2  is T, S, A or N and X 3  is G or A (SEQ ID NO:57, provided that the amino acid sequence is not QSLLDSDGKTY (SEQ ID NO: 36) or QSLVDSDGKTY (SEQ ID NO: 37); and   b) CDR3 of VH set forth in SEQ ID NO: 3 and CDR3 of VL set forth in SEQ ID NO: 5.   
     
     
         3 . The antibody or binding fragment thereof of  claim 2 , wherein the antibody or binding fragment thereof comprises:
 a) CDR1 of VL, wherein the CDR comprises or consists of the amino acid sequence QSL-X 1 -D-X 2 -D-X 3 -KTY, wherein X 1  is V or L, X 2  is T, S, A or N and X 3  is G or A (SEQ ID NO:57, provided that the amino acid sequence is not QSLLDSDGKTY (SEQ ID NO: 36) or QSLVDSDGKTY (SEQ ID NO: 37); and   b) CDR1 of VH set forth in SEQ ID NO:1, CDR2 of VH set forth in SEQ ID NO:2, CDR3 of VH set forth in SEQ ID NO:3, CDR2 of VL set forth in SEQ ID NO:4, and CDR3 of VL set forth in SEQ ID NO:5.   
     
     
         4 . The antibody or binding fragment thereof of  claim 1 , wherein the antibody or binding fragment thereof comprises:
 a) CDR1 of VL selected from CDR1 of VL set forth in one of SEQ ID NOs: 6, 7, 8, 9 and 10;   b) CDR1 of VH set forth in SEQ ID NO:1, CDR2 of VH set forth in SEQ ID NO:2, CDR3 of VH set forth in SEQ ID NO:3, CDR2 of VL set forth in SEQ ID NO:4, and CDR3 of VL set forth in SEQ ID NO:5.   
     
     
         5 . The antibody or binding fragment thereof of  claim 1 , wherein the antibody or binding fragment thereof comprises:
 a) CDR1 of VL, wherein the CDR comprises or consists of the amino acid sequence QSL-X 1 -D-X 2 -D-X 3 -KTY, wherein X 1  is V or L, X 2  is T, S, A or N and X 3  is G or A (SEQ ID NO:57, provided that the amino acid sequence is not QSLLDSDGKTY (SEQ ID NO: 36) or QSLVDSDGKTY (SEQ ID NO: 37);   b) at least one of: CDR2 of VL set forth in SEQ ID NO:4 and CDR3 of VL set forth in SEQ ID NO:5; and   c)
 (i) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11 or 12; or 
 (ii) a fragment of at least 7 amino acids of (i), wherein the antibody or binding fragment thereof retains the ability of being specifically reactive with a citrullinated epitope on deiminated human histone 2A and/or histone 4; or 
 (iii) a variant of (i) having at least 70% amino acid sequence identity to a sequence of (i), wherein the antibody or binding fragment thereof retains the ability of being specifically reactive with a citrullinated epitope on deiminated human histone 2A and/or histone 4. 
   
     
     
         6 . The antibody or binding fragment thereof of  claim 5 , wherein the antibody or binding fragment thereof comprises:
 a) CDR1 of VL selected from CDR1 of VL set forth in one of SEQ ID NOs: 6, 7, 8, 9 and 10;   b) CDR2 of VL set forth in SEQ ID NO:4 and CDR3 of VL set forth in SEQ ID NO:5; and   c) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11 or 12.   
     
     
         7 . The antibody or binding fragment thereof of  claim 1 , wherein the antibody or binding fragment thereof comprises:
 a) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 13;   b) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 14;   c) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 15;   d) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 16;   e) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 17;   f) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 12 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 13;   g) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 12 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 14;   h) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 12 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 15;   i) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 12 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 16; or   j) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 12 and the light chain variable domain amino acid sequence set forth in SEQ ID NO: 17.   
     
     
         8 . The antibody or binding fragment thereof of  claim 1  that specifically binds to a citrullinated epitope on deiminated human histone 2A and/or histone 4, wherein the antibody or binding fragment thereof comprises:
 a) CDR1 present in one of SEQ ID NOs: 13, 14, 15, 16 and 17; and 
 b) the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11 or 12. 
 
     
     
         9 . The antibody or binding fragment thereof of  claim 1 , that specifically binds to a peptide selected from the group consisting of SEQ ID NOs: 18, 19, 20, 21 and 22, and binds deiminated human histone 2A and/or histone 4. 
     
     
         10 . The antibody or binding fragment thereof of  claim 1 , that specifically binds to a citrullinated epitope on deiminated human histone 2A and/or histone 4 with an affinity of at least 1 nM or less. 
     
     
         11 . The antibody or binding fragment thereof of  claim 1 , wherein the antibody or binding fragment thereof is selected from the group consisting of recombinant antibodies, single chain antibodies, single chain variable fragments (scFv), variable fragments (Fv), fragment antigen-binding regions (Fab), single-domain antibodies (sdAb), VHH antibodies, nanobodies, camelids-derived single-domain antibodies, shark IgNAR-derived single-domain antibody fragments (VNAR), diabodies, triabodies, Anticalins and aptamers, or a full length antibody, optionally comprising an Fc region, such as an IgG1, IgG2, IgG3 or IgG4 Fc region. 
     
     
         12 .- 13 . (canceled) 
     
     
         14 . The antibody or binding fragment thereof of  claim 11 , comprising a heavy chain constant region comprising SEQ ID NO: 23 or 56, and/or a light chain constant region comprising SEQ ID NO: 24. 
     
     
         15 . The antibody or binding fragment thereof of  claim 14 , wherein the antibody comprises the heavy chain variable domain amino acid sequence set forth in SEQ ID NO: 11, the light chain variable domain amino acid sequence set forth in SEQ ID NO: 16, the heavy chain constant region amino acid sequence set forth in SEQ ID NO: 23 or 56, and the light chain constant region amino acid sequence set forth in SEQ ID NO: 24. 
     
     
         16 . The antibody or binding fragment thereof of  claim 1 , that is conjugated to an additional moiety. 
     
     
         17 . A polynucleotide encoding the antibody or binding fragment thereof of  claim 1  . 
     
     
         18 . A process for the production of an antibody or binding fragment thereof that specifically binds to a citrullinated epitope on deiminated human histone 2A and/or histone 4, comprising culturing a host cell expressing the antibody or binding fragment thereof of  claim 1 , and isolating the antibody or binding fragment thereof from said cell. 
     
     
         19 . A pharmaceutical composition comprising the antibody or binding fragment thereof of  claim 1 , and at least one pharmaceutically acceptable diluent or carrier. 
     
     
         20 . The pharmaceutical composition of  claim 19 , further comprising another active ingredient. 
     
     
         21 .- 24 . (canceled) 
     
     
         25 . A method of treating a patient, comprising administering a therapeutically effective amount of the antibody or binding fragment thereof of  claim 1 , to said patient,
 optionally wherein the treatment is of a NET-associated pathology, which is preferably systemic lupus erythematosus (SLE), lupus, sepsis, vasculitis, inflammatory arthritis, rheumatoid arthritis and osteoarthritis, psoriasis, Alzheimer's disease, autoimmune hepatitis, juvenile idiopathic arthritis, Sjögren's disease, Anti-phospholipid Syndrome, Bechet's disease, spondylitis, spondyloarthropathy, multiple system atrophy, Parkinson's disease, Lewy body dementia asthma, allergic rhinovirus exacerbated asthma, allergic asthma, cystic fibrosis, fibrosis and idiopathic pulmonary fibrosis, dry eye disease, uveitis, nongranulomatous uveitis, granulomatous uveitis, dermatitis, atopic dermatitis, COPD, bronchitis, or another NET-associated pathology such as wound healing in diabetes, cancer, cancer metastasis, and transplant organ health in vivo or ex vivo;   and optionally wherein administration is by a parenteral route of administration such as an intravenous, subcutaneous, intraocular, intramuscular, intradermal, intraperitoneal, spinal route or by injection or infusion; or by another administration route such as rectal, oral, ocular, topical, epidermal, mucosal, local, peritumoral, juxtatumoral, intratumoral, to the resection margin of tumors, intralesional, perilesional, by intra cavity infusion, intravesicle administration, or by inhalation.   
     
     
         26 . (canceled) 
     
     
         27 . A cloning or expression vector comprising the polynucleotide of  claim 17 . 
     
     
         28 . A host cell comprising the cloning or expression vector of  claim 27 .