HUMANIZED ANTIBODIES THAT BIND LGR5
Disclosed herein are humanized anti-LGR5 antibodies for the treatment of cancer. Antibodies disclosed herein may bind LGR5 without disrupting LGR5-RSPO1 binding or signaling, and may disrupt LGR5 signaling through Wnt that is independent of RSPO1. Also disclosed are heavy and light chain polypeptide sequences for the biding of LGR5, for example without disrupting LGR5-RSPO binding or signaling.
1 .- 71 . (canceled)
72 . A human or humanized antibody or epitope-binding fragment thereof that specifically binds human leucine-rich repeat containing G-protein-coupled receptor 5 (LGR5) comprising:
a heavy chain complementary determining region 1 (CDR1) comprising an amino acid sequence as shown in SEQ ID NO:23, or conservative variations thereof;
a heavy chain complementary determining region 2 (CDR2) comprising an amino acid sequence as shown in SEQ ID NO:25;
a heavy chain complementary determining region 3 (CDR3) comprising an amino acid sequence as shown in SEQ ID NO:27;
a light chain CDR1 comprising an amino acid sequence as shown in SEQ ID NO:29;
a light chain CDR2 comprising an amino acid sequence as shown in SEQ ID NO:31; and
a light chain CDR3 comprising an amino acid sequence as shown in SEQ ID NO:33.
73 . The human or humanized antibody or epitope-binding fragment thereof of claim 72 , wherein the heavy chain CDR1 comprises an amino acid sequence as shown in SEQ ID NO:23 with conservative substitutions thereof.
74 . The human or humanized antibody or epitope-binding fragment thereof of claim 72 , wherein the heavy chain CDR1 comprises a polypeptide having at least 90% identity with an amino acid sequence as shown in SEQ ID NO:23.
75 . The human or humanized antibody or epitope-binding fragment thereof of claim 72 , wherein the heavy chain CDR1 comprises one or more conservative substitutions at a residue as shown in SEQ ID NO:23 and selected from the group consisting of the third residue, the fifth residue, and the sixth residue.
76 . The human or humanized antibody or epitope-binding fragment thereof of claim 72 , wherein the heavy chain CDR1 comprises an amino acid sequence as shown in SEQ ID NO:23.
77 . The human or humanized antibody of claim 72 comprising:
a heavy chain variable domain comprising SEQ ID NO: 19; or
a light chain variable domain comprising SEQ ID NO: 21.
78 . The human or humanized antibody or epitope-binding fragment thereof of claim 72 comprising:
a heavy chain comprising SEQ ID NOs: 13; or
a light chain comprising SEQ ID NOs: 14.
79 . The human or humanized antibody or epitope-binding fragment thereof of claim 72 comprising activity to inhibit growth of a human colon tumor cell in vivo.
80 . The human or humanized antibody or epitope-binding fragment thereof of claim 79 , wherein the human colon tumor cell is a cancer stem cell.
81 . The human or humanized antibody or epitope-binding fragment thereof of claim 80 , wherein the cancer stem cell expresses a CD44 or a CD166 marker on its surface.
82 . The humanized antibody of or epitope-binding fragment thereof claim 72 , comprising activity to inhibit growth of a human neoplastic cell in vivo, wherein the neoplastic cell is selected from the group consisting of a lung tumor cell, a breast tumor cell, a colon tumor cell, and a pancreatic tumor cell.
83 . The human or humanized antibody or epitope-binding fragment thereof of claim 72 comprising activity to inhibit growth of a human neoplastic cell in vivo, wherein the neoplastic cell is selected from the group consisting of a triple negative breast cancer cell, a colon cancer cell having a mutation in a gene selected from the group consisting of K-Ras, H-Ras, APC, PI3K, PTEN, p53, STK11, RB1, TP53, FGFR2, VANGL2, and ISCO, and a small cell lung cancer cell.
84 . An epitope-binding fragment of the human or humanized antibody of claim 72 .
85 . The human or humanized antibody or epitope-binding fragment thereof of claim 72 in a lyophilized form.
86 . A pharmaceutical composition comprising the human or humanized or epitope-binding fragment thereof antibody of claim 72 and a pharmaceutically acceptable carrier.
87 . The pharmaceutical composition of claim 86 suitable for intravenous administration.
88 . A kit comprising the human or humanized antibody or epitope-binding fragment thereof of claim 72 and a chemotherapeutic agent.
89 . The kit of claim 88 , wherein the chemotherapeutic agent is selected from the group consisting of folinic acid, fluorouracil, irinotecan, gemcitabine and nab-paclitaxel.