Novel antibodies against cancer target block tumor growth, angiogenesis and metastasis

Abstract

The present invention provides antibodies that target the first-third domains of N-cadherein and the fourth domain of N-cadherin, for diagnosis and therapy of cancers related to N-cadherein. Methods of diagnosis and treatment utilizing these antibodies are also described.

1 . A hybridoma cell line deposited as ATCC Accession No. PTA-9387. 
     
     
         2 . An antibody produced by the hybridoma cell line of  claim 1 . 
     
     
         3 . A hybridoma cell line deposited as ATCC Accession No. PTA-9388. 
     
     
         4 . An antibody produced by the hybridoma cell line of  claim 3 . 
     
     
         5 . An antibody or fragment thereof capable of binding to domains 1-3 of N-cadherin, the same antigenic determinant of N-cadherin, in vivo or in vitro, as does the monoclonal antibody produced by the hybridoma cell line deposited at the American Type Culture Collection having ATCC Accession No. PTA-9387. 
     
     
         6 . The antibody of  claim 5 , wherein the antibody is humanized or fully human. 
     
     
         7 . The antibody of  claim 5 , wherein the antibody is a diabody or scFv. 
     
     
         8 . The antibody of  claim 5 , wherein the antibody comprises a heavy chain comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO:1. 
     
     
         9 . The antibody of  claim 5 , wherein the antibody comprises SEQ ID NO:1. 
     
     
         10 . The antibody of  claim 5 , wherein the antibody comprises a light chain comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO:2. 
     
     
         11 . The antibody of  claim 5 , wherein the antibody comprises SEQ ID NO:2. 
     
     
         12 . The antibody of  claim 5 , wherein the antibody comprises SEQ ID NO:1 and SEQ ID NO:2. 
     
     
         13 . An antibody or fragment thereof capable of binding to domain 4 of N-cadherin, the same antigenic determinant of N-cadherin, in vivo or in vitro, as does the monoclonal antibody produced by the hybridoma cell line deposited at the American Type Culture Collection having ATCC Accession No. PTA-9388. 
     
     
         14 . The antibody of  claim 13 , wherein the antibody is humanized or fully human. 
     
     
         15 . The antibody of  claim 13 , wherein the antibody is a diabody or scFv. 
     
     
         16 . The antibody of  claim 13 , wherein the antibody comprises a heavy chain comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO:3. 
     
     
         17 . The antibody of  claim 13 , wherein the antibody comprises SEQ ID NO:3. 
     
     
         18 . The antibody of  claim 13 , wherein the antibody comprises a light chain comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO:4. 
     
     
         19 . The antibody of  claim 13 , wherein the antibody comprises SEQ ID NO:4. 
     
     
         20 . The antibody of  claim 13 , wherein the antibody comprises SEQ ID NO:3 and SEQ ID NO:4. 
     
     
         21 . A method of inhibiting the growth of cancer cells in a patient, the method comprising the step of:
 administering an antibody or fragment thereof of  claim 5  or  13  to a patient under conditions sufficient for binding the antibody or fragment thereof to said cancer cells, wherein the cancer cells express or overexpress N-Cadherin, and wherein the antibody inhibits the growth of cancer cells by
 (a) activating or inhibiting NF kappa-B signaling and transcription; 
 (b) activating or inhibiting N-cadherin internalization; 
 (c) activating or inhibiting PI3 kinase or Akt pathway; 
 (d) activating or inhibiting β-catenin signaling; 
 (e) blocking heterodimerization of N-cadherin with FGFR or other tyrosine kinase receptor; or 
 (f) blocking or enhancing cleavage by ADAM10 or other metallopeptidase. 
   
     
     
         22 . The method of  claim 21 , wherein the cancer cells are urogenital cancer cells. 
     
     
         23 . The method of  claim 21 , wherein the cancer cells are prostate cancer cells. 
     
     
         24 . The method of  claim 21 , wherein the cancer cells are bladder cancer cells. 
     
     
         25 . A method of treating a cancer patient, comprising the steps of:
 (a) obtaining a test tissue sample from an individual at risk of having a cancer that expresses a N-cadherin protein;   (b) determining the presence or absence or amount of the N-cadherin protein in the test tissue sample in comparison to a control tissue sample from an individual known to be negative for the cancer; thereby diagnosing said cancer that expresses a N-cadherin protein, wherein the N-cadherin protein is expressed at normal or low levels, or is expressed by a subset of cells, or is overexpressed; and   (c) administering an effective amount of N-cadherin antibody or fragment of  claim 5  or  13  to the individual at risk of having a cancer that expresses a N-Cadherin protein.   
     
     
         26 . The method of  claim 25 , wherein said tissue sample is prostate or bladder tissue. 
     
     
         27 . The method of  claim 25 , wherein said cancer is a prostate cancer. 
     
     
         28 . The method of  claim 25 , wherein said cancer is a bladder cancer. 
     
     
         29 . The method of  claim 25 , wherein said antibody blocks hormone refractory prostate cancer. 
     
     
         30 . The method of  claim 25 , wherein said antibody blocks cancer stem cells. 
     
     
         31 . The method of  claim 25 , wherein said tissue sample is prostate or bladder tissue. 
     
     
         32 . The method of  claim 25 , wherein said cancer is a metastatic cancer. 
     
     
         33 . The method of  claim 25 , wherein said antibody is a monoclonal antibody. 
     
     
         34 . The method of  claim 25 , wherein the antibody is a scFv. 
     
     
         35 . The method of  claim 25 , wherein the antibody is a diabody. 
     
     
         36 . A method of diagnosing a cancer patient, comprising the steps of:
 (a) obtaining a test tissue sample from an individual at risk of having a cancer that expresses a N-cadherin protein;   (b) determining the presence or absence or amount of the N-cadherin protein in the test tissue sample in comparison to a control tissue sample from an individual known to be negative for the cancer by contacting a sample with an effective amount of N-cadherin antibody or fragment of  claim 5  or  13 ; thereby diagnosing said cancer that expresses a N-cadherin protein, wherein the N-cadherin protein is expressed at normal or low levels, or is expressed by a subset of cells, or is overexpressed.   
     
     
         37 . A method of identifying cancer stem cells, comprising the steps of:
 (a) obtaining a test tissue sample from an individual at risk of having a cancer that expresses a N-cadherin protein;   (b) determining the presence or absence of cancer stem cells in the test tissue sample in comparison to a control tissue sample from an individual known to be negative for the cancer; wherein the N-cadherin protein is expressed at normal or low levels, or is expressed by a subset of the stem cells and is not overexpressed, using the antibodies of  claim 5  or  13 .   
     
     
         38 . The method of  claim 37 , wherein said tissue sample is prostate or bladder tissue. 
     
     
         39 . The method of  claim 37 , wherein said cancer is a prostate cancer. 
     
     
         40 . The method of  claim 37 , wherein said cancer is a bladder cancer. 
     
     
         41 . The method of  claim 37 , wherein said cancer is a hormone refractory prostate cancer. 
     
     
         42 . The method of  claim 37 , wherein said cancer is a metastatic cancer.