Selectin ligands useful in the diagnosis and treatment of cancer

Abstract

The present invention provides methods and compositions useful in the diagnosis and treatment of cancer. More specifically, the present invention provides compositions and methods of use comprising a targeting composition comprising a solid substrate, an antibody composition, and optionally a chemotherapeutic agent.

1 . A targeting composition comprising:
 a. a solid substrate;   b. an antibody composition bound to the substrate, comprising at least one antibody to one or more E-selectin and L-selectin ligand antigens, the antigens comprising podocalyxin-like protein (PCLP) or two or more of PCLP, carcinoembryonic antigen (CEA), and CD44v, the antibody composition binding specifically to metastatic tumor cells having E-selectin and L-selectin binding activity,   wherein the substrate is a patterned array and/or the composition further comprises a therapeutic or imaging agent bound to the substrate or antibody.   
     
     
         2 . The targeting composition of  claim 1 , wherein the substrate is a particle suspendable in a biocompatible medium. 
     
     
         3 . The targeting composition of  claim 1 , wherein the particle is selected from the group consisting of a nanoparticle, a quantum dot, a liposome, a micelle and a polymersome. 
     
     
         4 . The targeting composition of  claim 1 , wherein the substrate is an array. 
     
     
         5 . The composition of  claim 1 , wherein the at least one antibody is selected from the group consisting of a polyclonal antibody, a monoclonal antibody, a recombinant antibody, a humanized antibody, a single chain antibody, and a Fab fragment. 
     
     
         6 . The composition of  claim 1 , wherein the antibody composition comprises an antibody to CEA, an antibody to PCLP, and an antibody to CD44v. 
     
     
         7 . The composition of  claim 1 , wherein the antibody composition comprises an antibody to PCLP and an antibody to CEA. 
     
     
         8 . The composition of  claim 1 , wherein the antibody composition comprises an antibody to PCLP and CD44v. 
     
     
         9 . The composition of  claim 1 , wherein the antibody composition comprises an antibody to CEA and CD44v. 
     
     
         10 . The composition of  claim 1 , wherein each of the at least one antibody is conjugated to the substrate via at least one linker molecule. 
     
     
         11 . The composition of  claim 1 , wherein the substrate is a nanoparticle. 
     
     
         12 . The composition of  claim 1 , wherein the substrate is a quantum dot. 
     
     
         13 . The composition of  claim 11 , wherein the nanoparticle is a gold nanoparticle. 
     
     
         14 . A method for treating cancer comprising administering to a subject with cancer a therapeutically effective amount of a composition comprising the targeting composition of  claim 1 , wherein the substrate is a particle suspendable in biocompatible medium, and further comprises at least one chemotherapeutic agent conjugated to the suspendable particle. 
     
     
         15 . The method of  claim 14 , wherein the cancer is a primary tumor. 
     
     
         16 . The method of  claim 15 , wherein the targeting composition prevents migration of the cancer from the primary tumor site. 
     
     
         17 . The method of  claim 14 , wherein the cancer is a metastatic cancer. 
     
     
         18 . The method of  claim 14 , wherein the targeting composition prevents formation of a secondary tumor. 
     
     
         19 . The method of  claim 14 , wherein the at least one antibody is selected from the group consisting of a polyclonal antibody, a monoclonal antibody, a recombinant antibody, a humanized antibody, a single chain antibody, and a Fab fragment. 
     
     
         20 . The method of  claim 14 , wherein the antibody composition comprises an antibody to CEA, an antibody to PCLP, and an antibody to CD44v. 
     
     
         21 . The composition of  claim 14 , wherein the antibody composition comprises an antibody to PCLP and an antibody to CEA. 
     
     
         22 . The composition of  claim 14 , wherein the antibody composition comprises an antibody to PCLP and CD44v. 
     
     
         23 . The composition of  claim 14 , wherein the antibody composition comprises an antibody to CEA and CD44v. 
     
     
         24 . The method of  claim 14 , the particle is a nanoparticle, a quantum dot, a liposome, a micelle and a polymersome. 
     
     
         25 . The method of  claim 14 , wherein the at least one antibody is conjugated to the particle via at least one linker molecule. 
     
     
         26 . The method of  claim 14 , wherein the particle is a quantum dot. 
     
     
         27 . The method of  claim 14 , wherein the particle is a gold nanoparticle. 
     
     
         28 . The method of  claim 14 , wherein the at least one chemotherapeutic agent is selected from the group consisting of paclitaxel, docetaxel, daunorubicin, cisplatin, carboplatin, oxaliplatin, colchicine, dolastatin 15, nocodazole podophyllotoxin, rhizoxin, vinblastine, vindesine, vinorelbine (navelbine), the epothilones, the mitomycins, bleomycin chlorambucil, cannustine, melphalan, mitoxantrone 5-fluoro-5′-deoxyuridine, camptothecin, topotecan, irinotecanetoposide, tenoposide, geldanamycin, methotrexate, adriamycin, actinomycin D, mifepristone, raloxifene, 5-azacytidine, 5-aza-2′-deoxycytidine, zebularine, tamoxifen, 4-hydroxytamoxifen apigenin, rapamycin, angiostatin K1-3, staurosporine, genistein, fumagillin, endostatin, thalidomide, analogs thereof and combinations thereof. 
     
     
         29 . A method for imaging a cancer cell in a patient comprising:
 a. administering to a subject a pharmaceutically acceptable composition comprising the targeting composition of  claim 1 , wherein the substrate is at least one particle suspendable in a biocompatible medium, and further comprises at least one imaging agent conjugated to the at least one particle; and   b. detecting the at least one imaging agent.   
     
     
         30 . The method of  claim 29 , wherein the imaging agent is selected from the group consisting of a radiologic contrast agent, diatrizoic acid sodium salt dihydrate, an iodine-containing agent, a barium-containing agent, a fluorescent imaging agent, Lissamine Rhodamine PE, a stain, a dye, a radioisotope, a metal, a ferromagnetic compound, a paramagnetic compound, gadolinium, a superparamagnetic compound, iron oxide, a diamagnetic compound, and barium sulfate. 
     
     
         31 . The method of  claim 29 , wherein the at least one antibody is selected from the group consisting of a polyclonal antibody, a monoclonal antibody, a recombinant antibody, a humanized antibody, a single chain antibody, and a Fab fragment. 
     
     
         32 . The method of  claim 29 , wherein the antibody composition comprises an antibody to CEA, an antibody to PCLP, and an antibody to CD44v. 
     
     
         33 . The composition of  claim 29 , wherein the antibody composition comprises an antibody to PCLP and an antibody to CEA. 
     
     
         34 . The composition of  claim 29 , wherein the antibody composition comprises an antibody to PCLP and CD44v. 
     
     
         35 . The composition of  claim 29 , wherein the antibody composition comprises an antibody to CEA and CD44v. 
     
     
         36 . The method of  claim 29 , the particle is a nanoparticle, a quantum dot, a liposome, a micelle and a polymersome. 
     
     
         37 . The method of  claim 29 , wherein the at least one antibody is conjugated to the particle via at least one linker molecule. 
     
     
         38 . The method of  claim 29 , wherein the particle is a quantum dot. 
     
     
         39 . The method of  claim 29 , wherein the article is a gold nanoparticle. 
     
     
         40 . The method of  claim 29 , wherein the detecting step comprises using an imaging device. 
     
     
         41 . An array comprising a fixed matrix and an antibody composition bound to the fixed matrix in a predetermined pattern, comprising at least one antibody to one or more E-selectin and L-selectin ligand antigens, the antigens comprising at least one of podocalyxin-like protein (PCLP), carcinoembryonic antigen (CEA), and CD44v, the antibody composition binding specifically to metastatic tumor cells having E-selectin and L-selectin binding activity. 
     
     
         42 . A method for diagnosing metastatic cancer comprising obtaining a biological sample from a patient and contacting the sample with an array of  claim 41 , wherein the specific binding by the targeting composition indicates the presence of metastatic cancer cells. 
     
     
         43 . The method of  claim 42 , wherein the array is a microarray. 
     
     
         44 . The method of  claim 42 , wherein the biological sample is blood. 
     
     
         45 . A method for preparing a targeting composition for metastatic tumor cells comprising binding an antibody composition to a substrate, the substrate being selected from a particle suspendable in a biocompatible medium and an array, and the antibody composition comprising at least one antibody to an E-selectin and L-selectin ligand antigen. 
     
     
         46 . The method of  claim 42 , wherein the antigen is selected from the group consisting of CEA, PCLP and CD44v. 
     
     
         47 . A composition comprising an antibody to PCLP, CEA, and/or CD44v conjugated with an imaging agent. 
     
     
         48 . The composition of  claim 47 , wherein the imaging agent is a radioactive isotope. 
     
     
         49 . The composition of  claim 48 , wherein the radioactive isotope is Iodine 131 . 
     
     
         50 . The composition of  claim 47 , wherein the antibody is selected from the group consisting of a polyclonal antibody, a monoclonal antibody, a recombinant antibody, a humanized antibody, a single chain antibody, and a Fab fragment. 
     
     
         51 . The composition of  claim 47 , wherein the antibody is to PCLP. 
     
     
         52 . A method for imaging a cancer cell in a patient comprising:
 a. administering to a subject the composition of  claim 47 ; and   b. detecting the at least one imaging agent.   
     
     
         53 . The method of  claim 52 , wherein the imaging agent is selected from the group consisting of a radiologic contrast agent, diatrizoic acid sodium salt dihydrate, an iodine-containing agent, a barium-containing agent, a fluorescent imaging agent, Lissamine Rhodamine PE, a stain, a dye, a radioisotope, a metal, a ferromagnetic compound, a paramagnetic compound, gadolinium, a superparamagnetic compound, iron oxide, a diamagnetic compound, and barium sulfate. 
     
     
         54 . The method of  claim 52 , wherein the antibody is selected from the group consisting of a polyclonal antibody, a monoclonal antibody, a recombinant antibody, a humanized antibody, a single chain antibody, and a Fab fragment. 
     
     
         55 . The method of  claim 52 , wherein the detecting step comprises using an imaging device. 
     
     
         56 . A method of identifying a biomarker specific for metastatic cancer cells comprising:
 a. selecting a putative glycopolypeptide selectin ligand found in a lysate of carcinoma cells;   b. detecting selectin binding activity of the putative glycopolypeptide selectin ligand in a blot rolling assay under shear flow conditions;   c. identifying the putative glycopolypeptide ligand, and   d. measuring selectin binding activity of the identified glycopolypeptide selectin ligand in a cell-free flow-based adhesion study.   
     
     
         57 . The method of  claim 56 , further comprising obtaining an antibody specific to the glycopolypeptide selectin ligand, wherein the antibody is specific to metastatic cells of the carcinoma. 
     
     
         58 . The method of  claim 56 , wherein the carcinoma cells are from colon, breast, prostate, squamous, neural blastoma, pancreatic, or lung cancer. 
     
     
         59 . The method of  claim 56 , wherein the identifying step comprises immunoaffinity chromatography, sequencing of isolated polypeptide fragments, and proteomics analysis. 
     
     
         60 . The method of  claim 56 , further comprising preparing a derivative of the identified glycopolypeptide selectin ligand and retesting in a cell-free flow-based adhesion study. 
     
     
         61 . The method of  claim 60 , wherein the derivative is a glycoprotein, a peptide, or a peptidomimetic. 
     
     
         62 . The method of  claim 60 , wherein the identified glycopolypeptide selectin ligand is selected from PCLP, CEA, and CD44v. 
     
     
         63 . The method of  claim 57 , wherein the antibody is a monoclonal antibody.