Anti-lgr5 antibodies and uses thereof

Abstract

The invention provides anti-LgR5 antibodies and methods of using the same.

What is claimed is: 
     
         1 . An isolated antibody that binds to LgR5, wherein the antibody comprises:
 a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 12, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 13, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 14, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 9, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 10, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 11; or   b) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 18, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 19, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 20, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 15, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 16, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 17.   
     
     
         2 . The antibody of  claim 1 , comprising:
 a) (i) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 6; (ii) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 5; or (iii) a VH sequence as in (i) and a VL sequence as in (ii); or   b) (i) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8; (ii) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7; or (iii) a VH sequence as in (i) and a VL sequence as in (ii).   
     
     
         3 . The antibody of  claim 1  or  claim 2 , comprising a VH sequence of SEQ ID NO: 6 or SEQ ID NO: 8. 
     
     
         4 . The antibody of any one of the preceding claims, comprising a VL sequence of SEQ ID NO: 7 or SEQ ID NO: 9. 
     
     
         5 . An antibody comprising a VH sequence of SEQ ID NO: 6 and a VL sequence of SEQ ID NO: 5. 
     
     
         6 . An antibody comprising a VH sequence of SEQ ID NO: 8 and a VL sequence of SEQ ID NO: 7. 
     
     
         7 . The antibody of any one of the preceding claims, which is a monoclonal antibody. 
     
     
         8 . The antibody of  claim 7 , which is a mouse, rabbit, human, humanized, or chimeric antibody. 
     
     
         9 . The antibody of any one of  claims 1  to  8 , which is an IgG selected from IgG1, IgG2a, IgG2b, IgG3, and IgG4. 
     
     
         10 . Isolated nucleic acid encoding the antibody of any one of the preceding claims. 
     
     
         11 . A host cell comprising the nucleic acid of  claim 10 . 
     
     
         12 . A method of producing an antibody comprising culturing the host cell of  claim 11  so that the antibody is produced. 
     
     
         13 . An immunoconjugate comprising the antibody of any one of  claims 1  to  9  and a cytotoxic agent. 
     
     
         14 . A pharmaceutical formulation comprising the immunoconjugate of  claim 13  and a pharmaceutically acceptable carrier. 
     
     
         15 . The antibody of any one of  claims 1  to  9  conjugated to a label. 
     
     
         16 . The antibody of  claim 15 , wherein the label is a positron emitter. 
     
     
         17 . The antibody of  claim 16 , wherein the positron emitter is  89 Zr. 
     
     
         18 . A method of detecting human LgR5 in a biological sample comprising contacting the biological sample with the anti-LgR5 antibody of any one of  claims 1  to  9  and  15  to  17  under conditions permissive for binding of the anti-LgR5 antibody to human LgR5, and detecting whether a complex is formed between the anti-LgR5 antibody and human LgR5 in the biological sample. 
     
     
         19 . The method of  claim 18 , wherein the anti-LgR5 antibody is an antibody of  claim 4  or  claim 5 . 
     
     
         20 . The method of  claim 18  or  claim 19 , wherein the biological sample is a colon cancer sample, a colorectal cancer sample, small intestine cancer sample, endometrial cancer sample, pancreatic cancer sample, or ovarian cancer sample. 
     
     
         21 . A method for detecting a LgR5-positive cancer comprising (i) administering a labeled anti-LgR5 antibody to a subject having or suspected of having a LgR5-positive cancer, wherein the labeled anti-LgR5 antibody comprises the anti-LgR5 antibody of any one of  claims 1  to  9 , and (ii) detecting the labeled anti-LgR5 antibody in the subject, wherein detection of the labeled anti-LgR5 antibody indicates a LgR5-positive cancer in the subject. 
     
     
         22 . The method of  claim 21 , wherein the labeled anti-LgR5 antibody is an antibody of  claim 4  or  claim 5  that is labeled. 
     
     
         23 . The method of  claim 21  or  claim 22 , wherein the labeled anti-LgR5 antibody comprises an anti-LgR5 antibody conjugated to a positron emitter. 
     
     
         24 . The method of  claim 23 , wherein the positron emitter is  89 Zr. 
     
     
         25 . A method of identifying a cancer patient as having a LgR5-positive cancer, comprising contacting a cancer sample from the patient with the anti-LgR5 antibody of any one of  claims 1  to  9  and  15  to  17  under conditions permissive for binding of the anti-LgR5 antibody to human LgR5, and detecting whether a complex is formed between the anti-LgR5 antibody and human LgR5 in the cancer sample. 
     
     
         26 . The method of  claim 25 , wherein the cancer patient is identified as having a LgR5-positive cancer if a complex is between the anti-LgR5 antibody and human LgR5 in the cancer sample is detected. 
     
     
         27 . A method of selecting a cancer patient for treatment with an immunoconjugate comprising an anti-LgR5 antibody, comprising determining the level of LgR5 expression in a cancer sample from the patient using immunohistochemistry (IHC), wherein an elevated level of LgR5 expression indicates that the cancer patient is more likely to benefit from treatment with an immunoconjugate comprising an anti-LgR5 antibody. 
     
     
         28 . The method of  claim 27 , wherein an elevated level of LgR5 expression is 2+ or 3+ staining by IHC. 
     
     
         29 . The method of  claim 28 , wherein an elevated level of LgR5 expression is 3+ staining by IHC. 
     
     
         30 . The method of any one of  claims 27  to  29 , wherein IHC is performed using an antibody of any one of  claims 1  to  9  and  15  to  17 . 
     
     
         31 . A method of selecting a cancer patient for treatment with an immunoconjugate comprising an anti-LgR5 antibody, comprising contacting a cancer sample from the patient with the anti-LgR5 antibody of any one of  claims 1  to  9  and  15  to  17  under conditions permissive for binding of the anti-LgR5 antibody to human LgR5, and detecting whether a complex is formed between the anti-LgR5 antibody and human LgR5 in the cancer sample. 
     
     
         32 . The method of  claim 31 , wherein the cancer patient is selected if a complex is between the anti-LgR5 antibody and human LgR5 in the cancer sample is detected. 
     
     
         33 . A method of treating a cancer patient comprising administering to the patient a therapeutically effective amount of an immunoconjugate comprising an anti-LgR5 antibody, wherein a cancer sample from the patient has been determined to have an elevated level of LgR5 expression using immunohistochemistry (IHC). 
     
     
         34 . The method of  claim 33 , wherein an elevated level of LgR5 expression is 2+ or 3+ staining by IHC. 
     
     
         35 . The method of  claim 34 , wherein an elevated level of LgR5 expression is 3+ staining by IHC. 
     
     
         36 . The method of any one of  claims 33  to  35 , wherein IHC is performed using an antibody of any one of  claims 1  to  9  and  15  to  17 . 
     
     
         37 . The method of any one of  claims 27  to  36 , wherein the immunoconjugate comprises an anti-LgR5 antibody comprising:
 a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 55, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 56, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 57, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 52; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 53; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 54; or 
 b) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 73, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 74, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 75, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 70; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 72; or 
 c) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 79, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 80, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 81, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 76; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 77; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 78; or 
 d) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 85, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 86, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 87, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 82; HVR-L2 comprising the amino acid sequence of SEQ ID NO: 83; and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 84. 
 
     
     
         38 . The method of  claim 37 , wherein the anti-LgR5 antibody comprises:
 a) a VH sequence of SEQ ID NO: 33 and a VL sequence of SEQ ID NO: 32; or   b) a VH sequence of SEQ ID NO: 51 and a VL sequence of SEQ ID NO: 50.   
     
     
         39 . The method of any one of  claims 27  to  38 , wherein the immunoconjugate comprises an anti-LgR5 antibody conjugated to a cytotoxic agent. 
     
     
         40 . The method of any one of  claims 25  to  39 , wherein the cancer sample is a colon cancer sample, a colorectal cancer sample, small intestine cancer sample, endometrial cancer sample, pancreatic cancer sample, or ovarian cancer sample.