Anti-CD303 Monoclonal Antibodies

Abstract

The invention relates to chimeric or humanised anti-CD303 antibodies to nucleic acids coding for the heavy and light chains of these antibodies, expression vectors, host cells, transgenic non-human animals or transgenic plants expressing said antibodies, as well as to the uses thereof in the treatment or prevention of blastic plasmacytoid dendritic cell neoplasms (BPDCN) or inflammatory diseases, in particular autoimmune diseases, involving plasmacytoid dendritic cells.

1 .- 30 . (canceled) 
     
     
         31 . A monoclonal antibody directed against the ectodomain of human CD303 antigen (SEQ ID NO: 130), or a functional fragment or a derivative thereof, wherein:
 a) the monoclonal antibody competes for binding to the human CD303 antigen with at least one antibody selected from:
 i) an antibody whose heavy chain variable region comprises SEQ ID NO: 43 and whose light chain variable region comprises SEQ ID NO: 48; 
 ii) an antibody whose heavy chain variable region comprises SEQ ID NO: 44 and whose light chain variable region comprises SEQ ID NO: 49; 
 iii) an antibody whose heavy chain variable region comprises SEQ ID NO: 45 and whose light chain variable region comprises SEQ ID NO: 50; 
 iv) an antibody whose heavy chain variable region comprises SEQ ID NO: 46 and whose light chain variable region comprises SEQ ID NO: 51; 
 v) an antibody whose heavy chain variable region comprises SEQ ID NO: 47 and whose light chain variable region comprises SEQ ID NO: 52; and 
   b) the light and heavy chain constant regions of the monoclonal antibody are constant regions from a non-murine species.   
     
     
         32 . The antibody, functional fragment or derivative thereof according to  claim 31 , wherein its heavy chains comprise three CDR-H (heavy-chain CDR according to IMGT nomenclature) having the following amino acid sequences, or sequences having at least 80% identity with the following sequences, and its light chains comprise three CDR-L (light-chain CDR according to IMGT nomenclature) having the following amino acid sequences, or sequences having at least 80% identity with the following sequences:
 i) CDR1-H-family 1: SEQ ID NO: 1, CDR2-H-family 1: SEQ ID NO: 2, CDR3-H-family 1: SEQ ID NO: 3, CDR1-L-family 1: SEQ ID NO: 4, CDR2-L-family 1: SEQ ID NO: 5, CDR3-L-family 1: SEQ ID NO: 6; or   ii) CDR1-H-family 2: SEQ ID NO: 7, CDR2-H-family 2: SEQ ID NO: 8, CDR3-H-family 2: SEQ ID NO: 9, CDR1-L-family 2: SEQ ID NO: 10, CDR2-L-family 2: SEQ ID NO: 11, CDR3-L-family 2: SEQ ID NO: 12.   
     
     
         33 . The antibody, functional fragment or derivative thereof according to  claim 32 , wherein the heavy chains comprise three CDR-H (heavy-chain CDR according to IMGT nomenclature) having the following amino acid sequences, or sequences having at least 80% identity with the following sequences, and the light chains comprise three CDR-L (light-chain CDR according to IMGT nomenclature) having the following amino acid sequences, or sequences having at least 80% identity with the following sequences:
 i) CDR1-H-122A2: SEQ ID NO: 13, CDR2-H-122A2: SEQ ID NO: 14, CDR3-H-122A2: SEQ ID NO: 15, CDR1-L-122A2: SEQ ID NO: 16, CDR2-L-122A2: SEQ ID NO: 17, CDR3-L-122A2: SEQ ID NO: 18;   ii) CDR1-H-102E9: SEQ ID NO: 19, CDR2-H-102E9: SEQ ID NO: 20, CDR3-H-102E9: SEQ ID NO: 21, CDR1-L-102E9: SEQ ID NO: 22, CDR2-L-102E9: SEQ ID NO: 23, CDR3-L-102E9: SEQ ID NO: 24;   iii) CDR1-H-104C12: SEQ ID NO: 25, CDR2-H-104C12: SEQ ID NO: 26, CDR3-H-104C12: SEQ ID NO: 27, CDR1-L-104C12: SEQ ID NO: 28, CDR2-L-104C12: SEQ ID NO: 29, CDR3-L-104C12: SEQ ID NO: 30;   iv) CDR1-H-114D11: SEQ ID NO: 31, CDR2-H-114D11: SEQ ID NO: 32, CDR3-H-114D11: SEQ ID NO: 33, CDR1-L-114D11: SEQ ID NO: 34, CDR2-L-114D11: SEQ ID NO: 35, CDR3-L-114D11: SEQ ID NO: 36; or   v) CDR1-H-104E10: SEQ ID NO: 37, CDR2-H-104E10: SEQ ID NO: 38, CDR3-H-104E10: SEQ ID NO: 39, CDR1-L-104E10: SEQ ID NO: 40, CDR2-L-104E10: SEQ ID NO: 41, CDR3-L-104E10: SEQ ID NO: 42.   
     
     
         34 . The antibody, functional fragment or derivative thereof according to  claim 31 , wherein its heavy chains comprise a variable region having a sequence selected from SEQ ID NOs: 43 to 47, 131 to 133, and 138 to 140, or having a sequence with at least 80% identity with one of SEQ ID NOs: 43 to 47, 131 to 133, and 138 to 140. 
     
     
         35 . The antibody, functional fragment or derivative thereof according to  claim 34 , wherein its light chains comprise a variable region having a sequence selected from SEQ ID NOs: 48 to 52, 134 to 137, and 141 to 143, or having a sequence with at least 80% identity with one of SEQ ID NOs: 48 to 52, 134 to 137, and 141 to 143. 
     
     
         36 . The antibody, functional fragment or derivative thereof according to  claim 31 , wherein it is a chimeric antibody. 
     
     
         37 . The antibody, functional fragment or derivative thereof according to  claim 36 , wherein it is a chimeric antibody whose heavy and light chains constant region is of human origin. 
     
     
         38 . The antibody, functional fragment or derivative thereof according to  claim 37 , wherein it has heavy and light chains comprising the following amino acid sequences or sequences having at least 80% identity with the following sequences:
 i) antibody 122A2: heavy chain: SEQ ID NO: 55, light chain: SEQ ID NO: 60,   ii) antibody 102E9: heavy chain: SEQ ID NO: 56, light chain: SEQ ID NO: 61,   iii) antibody 104C12: heavy chain: SEQ ID NO: 57, light chain: SEQ ID NO: 62,   iv) antibody 114D11: heavy chain: SEQ ID NO: 58, light chain: SEQ ID NO: 63, or   v) antibody 104E10: heavy chain: SEQ ID NO: 59, light chain: SEQ ID NO: 64.   
     
     
         39 . The antibody, functional fragment or derivative thereof according to  claim 31 , wherein it is a humanized antibody. 
     
     
         40 . The antibody, functional fragment or derivative thereof according to  claim 39 , wherein its heavy chains comprise a variable region having a sequence selected from SEQ ID NOs: 131 to 133 and 138 to 140 or having a sequence with at least 80% identity with one of SEQ ID NOs: 131 to 133. 
     
     
         41 . The antibody, functional fragment or derivative thereof according to  claim 40 , wherein its light chains comprise a variable region having a sequence selected from SEQ ID NOs: 134 to 137 and 141 to 143 or having a sequence with at least 80% identity with one of SEQ ID NOs: 134 to 137 and 141 to 143. 
     
     
         42 . The antibody, functional fragment or derivative thereof according to  claim 39 , wherein it has heavy and light chains with variable regions having the following amino acid sequences or having sequences with at least 80% identity with the following sequences:
 i) antibody 122A2H5: heavy chain: SEQ ID NO: 131, light chain: SEQ ID NO: 134,   ii) antibody 122A2H6: heavy chain: SEQ ID NO: 132, light chain: SEQ ID NO: 134,   iii) antibody 122A2H7: heavy chain: SEQ ID NO: 133, light chain: SEQ ID NO: 134,   iv) antibody 122A2H9: heavy chain: SEQ ID NO: 131, light chain: SEQ ID NO: 135,   v) antibody 122A2H10: heavy chain: SEQ ID NO: 132, light chain: SEQ ID NO: 135,   vi) antibody 122A2H14: heavy chain: SEQ ID NO: 132, light chain: SEQ ID NO: 136,   vii) antibody 102E9H6: heavy chain: SEQ ID NO: 139, light chain: SEQ ID NO: 141,   viii) antibody 102E9H7: heavy chain: SEQ ID NO: 140, light chain: SEQ ID NO: 141,   ix) antibody 102E9H9: heavy chain: SEQ ID NO: 138, light chain: SEQ ID NO: 142,   x) antibody 102E9H10: heavy chain: SEQ ID NO: 139, light chain: SEQ ID NO: 142.   
     
     
         43 . The antibody, functional fragment or derivative thereof according to  claim 42 , wherein it has heavy and light chains comprising the following amino acid sequences or sequences having at least 80% identity with the following sequences:
 i) antibody 122A2H5: heavy chain: SEQ ID NO: 150, light chain: SEQ ID NO: 153,   ii) antibody 122A2H6: heavy chain: SEQ ID NO: 151, light chain: SEQ ID NO: 153,   iii) antibody 122A2H7: heavy chain: SEQ ID NO: 152, light chain: SEQ ID NO: 153,   iv) antibody 122A2H9: heavy chain: SEQ ID NO: 150, light chain: SEQ ID NO: 154,   v) antibody 122A2H10: heavy chain: SEQ ID NO: 151, light chain: SEQ ID NO: 154,   vi) antibody 122A2H14: heavy chain: SEQ ID NO: 151, light chain: SEQ ID NO: 155,   vii) antibody 102E9H6: heavy chain: SEQ ID NO: 158, light chain: SEQ ID NO: 160,   viii) antibody 102E9H7: heavy chain: SEQ ID NO: 159, light chain: SEQ ID NO: 160,   ix) antibody 102E9H9: heavy chain: SEQ ID NO: 157, light chain: SEQ ID NO: 161,   x) antibody 102E9H10: heavy chain: SEQ ID NO: 158, light chain: SEQ ID NO: 161.   
     
     
         44 . The antibody, functional fragment or derivative thereof according to  claim 31 , wherein it is of IgG isotype. 
     
     
         45 . The antibody, functional fragment or derivative thereof according to  claim 31 , wherein its heavy chain and/or its light chain further comprise(s) a heterologous signal peptide of SEQ ID NO: 65. 
     
     
         46 . The antibody, functional fragment or derivative thereof according to  claim 31 , wherein it is produced in a cell line selected from: SP2/0; YB2/0; IR983F; human myeloma Namalwa; PERC6; CHO cell lines, notably CHO-K-1, CHO-Lec10, CHO-Lec1, CHO-Lec13, CHO Pro-5, CHO dhfr-, or the CHO cell line deleted for both alleles encoding the FUT8 gene and/or the GMD gene; Wil-2; Jurkat; Vero; MoIt-4; COS-7; 293-HEK; BHK; K6H6; NSO; SP2/0-Ag 14, P3X63Ag8.653, duck embryonic cell line EB66® (Valneva); rat hepatoma cell lines H4-II-E (DSM ACC3129) and H4-II-Es (DSM ACC3130), NM-H9D8 (DSM ACC2806), NM-H9D8-E6 (DSM ACC 2807) and NM H9D8-E6Q12 (DSM ACC 2856). 
     
     
         47 . The antibody, functional fragment or derivative thereof according to  claim 31 , having:
 a) fucose content less than or equal to 65%;   b) oligomannose-type N-glycans content greater than or equal to 30%; or   c) galactose content greater than or equal to 50%.   
     
     
         48 . The antibody, functional fragment or derivative thereof according to  claim 31 , wherein:
 a) the Fc fragment has the following mutations or combinations of mutations in their Fc fragment:
 N315D/A330V/N361D/A378V/N434Y, 
 P230S/N315D/M428L/N434Y, 
 E294del/T307P/N434Y, 
 T307A/N315D/A330V/E382V/N389T/N434Y, 
 V259I/N315D/N434Y, or 
 T256N/A378V/S383N/N434Y, 
   wherein the numbering of Fc fragment amino acids is that of the EU index of Kabat; or   b) it has a deletion of the amino acid at position 293 (Del293) or 294 (Del294) of the Fc fragment, where the numbering of Fc fragment amino acids is that of the EU index of Kabat.   
     
     
         49 . The functional fragment according to  claim 31 , wherein it is selected from the fragments Fv, ScFv, Fab, F(ab′)2, Fab′, scFv-Fc and diabodies. 
     
     
         50 . A nucleic acid encoding the heavy and/or light chain of an antibody, functional fragment or derivative thereof according to  claim 31 . 
     
     
         51 . The nucleic acid according to  claim 50 , comprising at least one of SEQ ID NOs: 86 to 95, and 181 to 193. 
     
     
         52 . A vector comprising a nucleic acid according to  claim 50 . 
     
     
         53 . A host cell, transgenic non-human animal or transgenic plant comprising at least one nucleic acid according to  claim 50  or a vector comprising the nucleic acid. 
     
     
         54 . The host cell according to  claim 53 , selected from the following lines: SP2/0; YB2/0; IR983F; human myeloma Namalwa; PERC6; CHO cell lines, notably CHO-K-1, CHO-Lec10, CHO-Lec1, CHO-Lec13, CHO Pro-5, CHO dhfr-, or the CHO cell line deleted for both alleles encoding the FUT8 gene and/or the GMD gene; Wil-2; Jurkat; Vero; MoIt-4; COS-7; 293-HEK; BHK; K6H6; NSO; SP2/0-Ag 14, P3X63Ag8.653, duck embryonic cell line EB66® (Valneva); rat hepatoma cell lines H4-II-E (DSM ACC3129) and H4-II-Es (DSM ACC3130), NM-H9D8 (DSM ACC2806), NM-H9D8-E6 (DSM ACC 2807) and NM H9D8-E6Q12 (DSM ACC 2856). 
     
     
         55 . A method for treating or preventing a hematopoietic tumor expressing the CD303 antigen in a patient, comprising administering to said patient an effective amount of an antibody, functional fragment or derivative thereof according to  claim 31 . 
     
     
         56 . A method for treating or preventing inflammatory disease in a patient, comprising administering to said patient an effective amount of an antibody, functional fragment or derivative thereof according to  claim 31 . 
     
     
         57 . A monoclonal antibody directed against the ectodomain of human CD303 antigen (SEQ ID NO: 130), or a functional fragment or a derivative thereof, wherein:
 the monoclonal antibody has improved affinity for Fey receptor Ma (FcγRIIIa, CD16a) compared to antibodies directed against the ectodomain of the human CD303 antigen, produced in CHO cells; and   the light and heavy chain constant regions of the monoclonal antibody are constant regions from a non-murine species.