HUMANIZED ANTIBODIES THAT BIND LGR5
Embodiments include humanized anti-LGR5 antibodies for the treatment of cancer. Some embodiments include antibodies which may bind LGR5 without disrupting LGR5-RSPO1 binding or signaling, and may disrupt LGR5 signaling through Wnt that is independent of RSPO1. Some embodiments include heavy and light chain polypeptide sequences for the binding of LGR5, for example without disrupting LGR5-RSPO binding or signaling.
1 . A human or humanized antibody or epitope-binding fragment thereof that specifically binds human leucine-rich repeat containing G-protein-coupled receptor 5 (LGR5) in a human LGR5 expressing tumor cell in vivo comprising:
a heavy chain complementarity determining region 1 (CDR1) comprising an amino acid sequence as shown in SEQ ID NO:23, or the amino acid sequence as shown in SEQ ID NO:23 comprising a substitution selected from (i) a serine to threonine substitution at residue 3, (ii) a threonine to serine substitution at residue 5, and (iii) an alanine to glycine substitution at residue 6;
a heavy chain complementarity determining region 2 (CDR2) comprising an amino acid sequence as shown in SEQ ID NO:25;
a heavy chain complementarity 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.
2 . The human or humanized antibody or epitope-binding fragment thereof of claim 1 , wherein the heavy chain CDR1 comprises the amino acid sequence as shown in SEQ ID NO:23 comprising no more than one substitution selected from (i) a serine to threonine substitution at residue 3, (ii) a threonine to serine substitution at residue 5, and (iii) an alanine to glycine substitution at residue 6.
3 . The human or humanized antibody or epitope-binding fragment thereof of claim 1 , wherein the heavy chain CDR1 comprises the amino acid sequence as shown in SEQ ID NO:23 comprising no more than two substitutions selected from (i) a serine to threonine substitution at residue 3, (ii) a threonine to serine substitution at residue 5, and (iii) an alanine to glycine substitution at residue 6.
4 . The human or humanized antibody or epitope-binding fragment thereof of claim 1 , wherein the heavy chain CDR1 comprises the amino acid sequence as shown in SEQ ID NO:23 having the conservative substitutions (i) a serine to threonine substitution at residue 3, (ii) a threonine to serine substitution at residue 5, and (iii) an alanine to glycine substitution at residue 6.
5 . The human or humanized antibody or epitope-binding fragment thereof of claim 1 , wherein the heavy chain CDR1 comprises an amino acid sequence as shown in SEQ ID NO:23.
6 . The human or humanized antibody of claim 1 comprising:
a heavy chain variable domain comprising SEQ ID NO: 19; or
a light chain variable domain comprising SEQ ID NO: 21.
7 . The human or humanized antibody or epitope-binding fragment thereof of claim 1 comprising:
a heavy chain comprising SEQ ID NOs: 13; or
a light chain comprising SEQ ID NOs: 14.
8 . The human or humanized antibody or epitope-binding fragment thereof of claim 1 comprising activity to inhibit growth of a human colon LGR5+ tumor cell in vivo, wherein the tumor cell is a cancer stem cell.
9 . The human or humanized antibody or epitope-binding fragment thereof of claim 8 , wherein the cancer stem cell expresses a CD44 or a CD166 marker on its surface.
10 . The humanized antibody of or epitope-binding fragment thereof claim 1 , comprising activity to inhibit growth of a human neoplastic LGR5+ cell in vivo, wherein the neoplastic cell is selected from the group consisting of a lung tumor cell, a breast tumor cell, a colon cancer cell, and a pancreatic tumor cell.
11 . The human or humanized antibody or epitope-binding fragment thereof of claim 1 comprising activity to inhibit growth of a human neoplastic LGR5+ cell in vivo, wherein the neoplastic cell is selected from the group consisting of a triple negative breast cancer cell, a colon cancer cell, and a small cell lung cancer cell, wherein the colon cancer cell comprises 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.
12 . An epitope-binding fragment of the human or humanized antibody of claim 1 .
13 . The human or humanized antibody or epitope-binding fragment thereof of claim 1 in a lyophilized form.
14 . A pharmaceutical composition comprising the human or humanized or epitope-binding fragment thereof antibody of claim 1 and a pharmaceutically acceptable carrier.
15 . The pharmaceutical composition of claim 14 suitable for intravenous administration.
16 . (canceled)
17 . (canceled)
18 . A method of treating a patient with a human LGR5 expressing tumor including the step of administering an effective amount of a human or humanized antibody or epitope-binding fragment thereof of claim 1 .
19 . A method of inhibiting a human LGR5 expressing tumor cell in vivo including the step of contacting said tumor cell with an effective amount of a human or humanized antibody or epitope-binding fragment thereof of claim 1 .