Compositions and methods for treatment and detection of cancers

Abstract

Pharmaceutical composition comprising antibodies or antigen binding fragments thereof that bind to SSEA-4 are disclosed herein, as well as methods of use thereof. Methods of use include, without limitation, cancer therapies and diagnostics. The antibodies of the disclosure can bind to certain cancer cell surfaces. Exemplary targets of the antibodies disclosed herein can include carcinomas, such as those in brain, lung, breast, mouse, esophagus, stomach, liver, bile duct, pancreas, colon, kidney, cervix, ovary, and/or prostate cancer.

1 . An isolated monoclonal antibody that specifically binds to Neu5Acα2→3Galβ1→3GalNAcβ1→3Galα1→4Galβ1→4Glcβ1. 
     
     
         2 . The isolated antibody of  claim 1 , which further binds to Fucα1→2Galβ1→3GalNAcβ1→3Galα1→4Galβ→4Glcβ1 and Galβ1→3GalNAcβ1→3Galα1→4Galβ1→4Glcβ1. 
     
     
         3 . The isolated antibody of  claim 1  or  2 , wherein the antibody is an IgG or IgM. 
     
     
         4 . The isolated monoclonal antibody of  claim 1 , wherein the antibody is not a mouse IgG3, and wherein the antibody is not a mouse IgM. 
     
     
         5 . The isolated antibody of  claim 4 , which further binds to Neu5Acα2→3Galβ1→3GalNAcβ1→3Galα1. 
     
     
         6 . The isolated antibody of  claim 4 , which further binds to Neu5Acα2→8→Neu5Acα2→3Galβ1→3GalNAcβ1→3Galα1→4Galβ1→4Glcβ1. 
     
     
         7 . The isolated antibody of  claim 1 , wherein the antibody is an IgG1. 
     
     
         8 . The isolated monoclonal antibody of  claim 1  which further binds to Neu5Acα2→3Galβ1→3GalNAcβ1→3Galα1, Fucα1→2Galβ1→3GalNAcβ1→3Galα1→4Galβ1→4Glcβ1, Galβ1→3GalNAcβ1→3Galα1→4Galβ1→4Glcβ1, GalNAcβ1→3Galα1→4Galβ1→4Glcβ1 and Neu5Acα2-8→Neu5Acα2→3Galβ1→3GalNAcβ1→3Galα1→4Galβ1→4Glcβ1. 
     
     
         9 . The isolated antibody of  claim 8 , wherein the antibody is an IgG or IgM. 
     
     
         10 . The isolated antibody of  claim 1 , wherein the antibody is a full-length antibody or an antigen-binding fragment thereof. 
     
     
         11 . The isolated antibody of  claim 10 , wherein the antigen binding fragment is a Fab fragment, a F(ab′)2 fragment, or a single-chain Fv fragment. 
     
     
         12 . The isolated antibody of  claim 1 , wherein antibody is a human antibody, a humanized antibody, a chimeric antibody, or a single-chain antibody. 
     
     
         13 . The isolated antibody of  claim 1 , or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises an H-CDR1, H-CDR2, and H-CDR3 selected from (i)-(iii) as set forth in Table I ( FIG. 16 ):
 (i) H-CDR1 selected from SEQ ID NO:52 or SEQ ID NO: 56;   (ii) H-CDR2 selected from of SEQ ID NO: 54 or SEQ ID NO: 58;   (iii) H-CDR3 selected from SEQ ID NO:60 or SEQ ID NO: 63, respectively;   and comprises an L-CDR1, L-CDR2 and L-CDR3 selected from (iv)-(vi):   (iv) L-CDR1 selected from SEQ ID NO: 66. or SEQ ID NO: 70;   (v) L-CDR2 selected from SEQ ID NO:68 or SEQ ID NO: 72; and   (vi) L-CDR3 selected from SEQ ID NO: 74, or SEQ ID NO: 77, respectively.   
     
     
         14 . The isolated antibody or antigen-binding fragment of  claim 13  wherein the antibody or the antigen binding fragment further comprising H-FR1, H-FR2, H-FR3, and HFR4 selected from (i)-(iv) as set forth in Table I ( FIG. 16 ):
 (i) H-FR1 selected from SEQ ID NO:51 or SEQ ID NO: 55; 
 (ii) H-FR2 selected from SEQ ID NO: 53 or SEQ ID NO: 57; 
 (iii) H-FR3 selected from SEQ ID NO:59 or SEQ ID NO: 62, 
 (iv) H-FR4 selected from SEQ ID NO:61 or SEQ ID NO: 64, respectively; 
 and comprising L-FR1, L-FR2, L-FR3 and L-FR4 selected from (v)-(viii): 
 (v) L-FR1 selected from SEQ ID NO: 65. or SEQ ID NO: 69; 
 (vi) L-FR2 selected from SEQ ID NO:67 or SEQ ID NO: 71; 
 (vii) L-FR3 selected from SEQ ID NO: 73, or SEQ ID NO: 76, 
 (viii) L-FR4 selected from SEQ ID NO: 75, or SEQ ID NO: 78, respectively. 
 
     
     
         15 . The isolated antibody or antigen-binding fragment of  claim 1  wherein the antibody or the antigen binding fragment optionally or substitutionally comprises an amino acid sequence that differs by less than three conservative amino acid substitutions from the amino acid sequence of any of SEQ ID NOs:51-78. 
     
     
         16 . The isolated antibody of  claim 1 , wherein the antibody comprises an anti SSEA4 humanized antibody or an antigen binding fragment thereof, comprising H-CDR1, HCDR2, and H-CDR3 selected from (i)-(iii) as set forth in Table II ( FIG. 16 ):
 (i) H-CDR1 having the sequence of SEQ ID NO:80;   (ii) H-CDR2 having the sequence of SEQ ID NO: 82,   (iii) H-CDR3 having the sequence SEQ ID NO:90 respectively;   and comprising L-CDR1, L-CDR2 and L-CDR3 selected from (iv)-(vi):   (iv) L-CDR1 having the sequence of SEQ ID NO: 99;   (v) L-CDR2 having the sequence of SEQ ID NO:101; and   (vi) L-CDR3 having the sequence of SEQ ID NO: 109, respectively.   
     
     
         17 . The isolated antibody or antigen-binding fragment of  claim 16  wherein the antibody or the antigen binding fragment further comprising H-FR1, H-FR2, H-FR3, and HFR4 selected from (i)-(iv) as set forth in Table I ( FIG. 16 ):
 (i) H-FR1 selected from SEQ ID NO:79, SEQ ID NO: 83, SEQ ID NO:85 or SEQ ID NO: 87; 
 (ii) H-FR2 selected from SEQ ID NO:81, SEQ ID NO: 84, SEQ ID NO:86 or SEQ ID NO: 88; 
 (iii) H-FR3 selected from SEQ ID NO:89, SEQ ID NO: 92, SEQ ID NO:94 or SEQ ID NO: 96; 
 (iv) H-FR4 selected from SEQ ID NO:91, SEQ ID NO: 93, SEQ ID NO:95 or SEQ ID NO: 97, respectively; and comprising L-FR1, L-FR2, L-FR3 and L-FR4 selected from (v)-(viii): 
 (v) L-FR1 selected from SEQ ID NO:98, SEQ ID NO: 102, SEQ ID NO:104 or SEQ ID NO: 106; 
 (vi) L-FR2 selected from SEQ ID NO:100, SEQ ID NO: 103, SEQ ID NO:105 or SEQ ID NO: 107; 
 (vii) L-FR3 selected from SEQ ID NO:108, SEQ ID NO: 111, SEQ ID NO:113 or SEQ ID NO: 114, 
 (viii) L-FR4 selected from SEQ ID NO:110 or SEQ ID NO: 112, respectively. 
 
     
     
         18 . The isolated antibody or antigen-binding fragment of  claim 1  wherein the antibody or the antigen binding fragment optionally or substitionally comprises an amino acid sequence that differs by less than ten conservative amino acid substitutions from the amino acid sequence of any of SEQ ID NOs:79-114. 
     
     
         19 . A method of treating cancer in a subject, the method comprising administering to a subject in need thereof an effective amount of the isolated antibody  claim 1 . 
     
     
         20 . The method of  claim 19 , wherein the cancer is selected from the group consisting of brain cancer, lung cancer, breast cancer, oral cancer, esophagus cancer, stomach cancer, liver cancer, bile duct cancer, pancreas cancer, colon cancer, kidney cancer, cervix cancer, ovarian cancer and prostate cancer. 
     
     
         21 . The method of  claim 19 , wherein the cancer is brain cancer, lung cancer, breast cancer, ovarian cancer, prostate cancer, colon cancer, or pancreas cancer. 
     
     
         22 . The method of  claim 19 , wherein the cancer is breast cancer, pancreas cancer, brain cancer or glioblastoma multiforme (GBM) cancer. 
     
     
         23 . The method of  claim 19 , wherein the subject is a human having or suspected of having cancer. 
     
     
         24 . The method of  claim 1 , further comprising administering an additional therapy to said subject prior to, during or subsequent to the administering of an isolated antibody of  claim 1 . 
     
     
         25 . The method of  claim 24 , wherein the additional therapy is treatment with a chemotherapeutic agent or radiation therapy. 
     
     
         26 . A method for detecting cancer in a subject, comprising:
 (a) applying one or more monoclonal antibodies that detect expression of a panel of markers consisting of GM3, GM2, GM1, GD1, GD1a, GD3, GD2, GT1b, A2B5, LeX, sLeX, LeY, SSEA-3, SSEA-4, sialyl SSEA-4, Globo H, Gb4, TF, Tn, sTn, CD44, CD24, CD45, CD90, CD133/1 and CD133/2 to a cell or tissue sample obtained from the subject;   (b) assaying the binding of the one or more monoclonal antibodies to the cell or the tissue sample; and   (c) comparing the binding with a normal control to determine the presence of the cancer in the subject.   
     
     
         27 . The method of  claim 26 , wherein the markers consist of GD2, GM2, GM1, GD1a, GT1b, A2B5, Tf, Tn, Globo H, Gb4, SSEA-3, SSEA-4, sialyl SSEA-4, CD24, CD44 and CD90. 
     
     
         28 . The method of  claim 26 , wherein the cells are selected from the group consisting of brain cancer cells, lung cancer cells, breast cancer cells, oral cancer cells, esophagus cancer cells, stomach cancer cells, liver cancer cells, bile duct cancer cells, pancreas cancer cells, colon cancer cells, kidney cancer cells, cervix cancer cells, ovarian cancer cells and prostate cancer cells. 
     
     
         29 . The method of  claim 28 , wherein the cancer is brain cancer glioblastoma multiforme (GBM) cancer, lung cancer, breast cancer, ovarian cancer, prostate cancer, colon cancer, pancreas cancer or other epithelial cancers. 
     
     
         30 . The method of  claim 29 , wherein the cancer is brain cancer or glioblastoma multiforme (GBM) cancer. 
     
     
         31 . A method for staging and/or determining prognosis of tumors in a human patient, the method comprising:
 (a) applying one or more monoclonal antibodies that detects one or more markers consisting of SSEA-3, SSEA-4 and Globo H to a cell or tissue sample obtained from the patient;   (b) assaying the binding of the monoclonal antibodies to the cell or the tissue sample;   (c) comparing the expression level of the markers in the test sample with the level in a reference sample, and   (d) determining the stage and/or prognosis of tumors in the patient based upon the outcome identified in step (c).   
     
     
         32 . The method of  claim 31 , wherein the tumor is brain tumor or glioblastoma multiforme (GBM). 
     
     
         33 . The method of  claim 31 , wherein the marker is SSEA-4. 
     
     
         34 . The method of  claim 33 , wherein the antibody further detects one or more of SSEA-3 or Globo H. 
     
     
         35 . The method of  claim 34 , wherein the one or more antibodies further comprise one or more antibodies that detect one or more of GM3, GM2, GM1, GD1, GD1a, GD3, GD2, GT1b, A2B5, LeX, sLeX, LeY, TF, Tn, sTn, CD44, CD24, CD45, CD90, CD133/1 and CD133/2 
     
     
         36 . A method for the isolation, enrichment, and self-renewal of stem cells from GBM tumor cells, using cell surface markers GD2, SSEA-4 and CD133. 
     
     
         37 . A method for isolating a cell population substantially enriched in tumorigenic stem cells derived from a GBM tumor. 
     
     
         38 . A pharmaceutical composition comprising the antibody or antigen binding fragment thereof according to  claim 1 . 
     
     
         39 . A method for administering an anti-SSEA4 antibody to a human in need thereof, the method comprising administering to said human from about 1 mg/kg to about 100 mg/kg of an anti-SSEA-4 antibody that demonstrates a binding affinity for SSEA-4.