Detection of Early-Stage Pancreatic Adenocarcinoma

Abstract

Described herein are compositions and methods of use of anti-pancreatic cancer antibodies or fragments thereof, such as murine, chimeric, humanized or human PAM4 antibodies. The subject antibodies show a number of novel and useful diagnostic characteristics, such as binding with high specificity to pancreatic and other cancers, but not to normal pancreatic tissues and binding to a high percentage of early stage pancreatic cancers. In preferred embodiments, the antibodies bind to pancreatic cancer mucins. The antibodies and fragments are of use for the detection and diagnosis of early stage pancreatic cancer. In preferred embodiments, the anti-pancreatic cancer antibodies can be used for immunoassay of serum samples, wherein the immunoassay can detect a marker for early stage pancreatic cancer in serum. More preferably, the serum is extracted with an organic phase, such as butanol, before immunoassay.

What is claimed is: 
     
         1 . A method of delivering a diagnostic or therapeutic agent to a pancreatic cancer cell comprising:
 a) obtaining an anti-MUC-5ac antibody or antigen-binding fragment thereof that binds to the same epitope as or competes for binding to pancreatic cancer mucin with a chimeric PAM4 (cPAM4) antibody that comprises the light chain variable region CDR sequences CDR1 (SASSSVSSSYLY, (SEQ ID NO:1), CDR2 (STSNLAS, SEQ ID NO:2), and CDR3 (HQWNRYPYT, SEQ ID NO:3) and the heavy chain variable region CDR sequences CDR1 (SYVLH, SEQ ID NO:4), CDR2 (YINPYNDGTQYNEKFKG, SEQ ID NO:5) and CDR3 (GFGGSYGFAY, SEQ ID NO:6), wherein the anti-MUC-5ac antibody or antigen-binding fragment thereof is conjugated to a diagnostic or therapeutic agent; and   b) exposing the pancreatic cancer cell to the conjugated anti-MUC-5ac antibody or antigen-binding fragment thereof.   
     
     
         2 . The method of  claim 1 , wherein the anti-MUC-5ac antibody or antigen-binding fragment thereof binds to early and late stage pancreatic cancer. 
     
     
         3 . The method of  claim 1 , wherein the anti-MUC-5ac antibody or antigen-binding fragment thereof binds to 80% or more of total pancreatic cancers. 
     
     
         4 . The method of  claim 1 , wherein the diagnostic agent is selected from the group consisting of a radionuclide, a contrast agent, a fluorescent agent, a chemiluminescent agent, a bioluminescent agent, a paramagnetic ion, an enzyme and a photoactive diagnostic agent. 
     
     
         5 . The method of  claim 4 , wherein the diagnostic agent is a radionuclide selected from the group consisting of  110 In,  111 In,  177 Lu,  18 F,  52 Fe,  62 Cu,  64 Cu,  67 Cu,  67 Ga,  68 Ga,  86 Y,  90 Y,  89 Zr,  94m Tc,  94 Tc,  99m Tc,  120 I,  123 I,  124 I,  125 I,  131 I,  154-158 Gd,  32 P,  11 C,  13 N,  15 O,  186 Re,  188 Re,  51 Mn,  52m Mn,  55 Co,  72 As,  75 Br,  76 Br,  82m Rb,  83 Sr, or other gamma-, beta-, or positron-emitters. 
     
     
         6 . The method of  claim 5 , wherein the radionuclide is  18 F. 
     
     
         7 . The method of  claim 4 , wherein the paramagnetic ion is selected from the group consisting of chromium (III), manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), gadolinium (III), vanadium (II), terbium (III), dysprosium (III), holmium (III) and erbium (III). 
     
     
         8 . The method of  claim 4 , wherein the diagnostic agent is a fluorescent labeling compound selected from the group consisting of fluorescein isothiocyanate, rhodamine, phycoerytherin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine, or a chemiluminescent labeling compound selected from the group consisting of luminol, isoluminol, an aromatic acridinium ester, an imidazole, an acridinium salt and an oxalate ester, or a bioluminescent compound selected from the group consisting of luciferin, luciferase and aequorin. 
     
     
         9 . The method of  claim 1 , wherein the therapeutic agent is selected from the group consisting of a radionuclide, an immunomodulator, a hormone, a hormone antagonist, an enzyme, an anti-sense oligonucleotide, an siRNA, an enzyme inhibitor, a photoactive therapeutic agent, a cytotoxic agent, a drug, a pro-drug, a toxin, an angiogenesis inhibitor and a pro-apoptotic agent. 
     
     
         10 . The method of  claim 9 , wherein the drug is selected from the group consisting of nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, gemcitabine, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzyme inhibitors, epipodophyllotoxins, platinum coordination complexes, vinca alkaloids, substituted ureas, methyl hydrazine derivatives, adrenocortical suppressants, hormone antagonists, endostatin, taxols, camptothecins, SN-38, doxorubicin, antimetabolites, alkylating agents, antimitotics, anti-angiogenic agents, tyrosine kinase inhibitors, mTOR inhibitors, heat shock protein (HSP90) inhibitors, proteosome inhibitors, HDAC inhibitors, pro-apoptotic agents, methotrexate and CPT-11. 
     
     
         11 . The method of  claim 9 , wherein the toxin is selected from the group consisting of ricin, abrin, alpha toxin, saporin, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin,  Pseudomonas  exotoxin, and  Pseudomonas  endotoxin. 
     
     
         12 . The method of  claim 9 , wherein the immunomodulator is selected from the group consisting of a cytokine, a stem cell growth factor, a lymphotoxin, a hematopoietic factor, a colony stimulating factor (CSF), an interleukin (IL), an interferon (IFN), a stem cell growth factor, erythropoietin, thrombopoietin, tumor necrosis factor (TNF), granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon-α, interferon-β, interferon-γ and the stem cell growth factor designated “S1 factor”. 
     
     
         13 . The method of  claim 12 , wherein the cytokine is selected from the group consisting of human growth hormone, N-methionyl human growth hormone, bovine growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, glycoprotein follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), placenta growth factor (PlGF), hepatic growth factor, prostaglandin, fibroblast growth factor, prolactin, placental lactogen, OB protein, tumor necrosis factor-α, tumor necrosis factor-β, mullerian-inhibiting substance, mouse gonadotropin-associated peptide, inhibin, activin, vascular endothelial growth factor, integrin, thrombopoietin (TPO), NGF-β, platelet-growth factor, TGF-α, TGF-β, insulin-like growth factor-I, insulin-like growth factor-II, erythropoietin (EPO), osteoinductive factors, interferon-α, interferon-β, interferon-γ, macrophage-CSF (M-CSF), IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, IL-25, LIF, FLT-3, angiostatin, thrombospondin, endostatin, TNF-α and LT (lymphotoxin). 
     
     
         14 . The method of  claim 9 , wherein the radionuclide is selected from the group consisting of  111 In,  177 Lu,  212 Bi,  213 Bi,  211 At,  62 Cu,  67 Cu,  90 Y,  125 I,  131 I,  32 P,  33 P,  47 Sc,  111 Ag,  67 Ga,  143 Pr,  153 Sm,  161 Tb,  166 Dy,  166 Ho,  186 Re,  188 Re,  189 Re,  212 Pb,  223 Ra,  225 Ac,  59 Fe,  75 Se,  77 As,  89 Sr,  99 Mo,  105 Rh,  109 Pd,  143 Pr,  149 Pm,  169 Er,  194 Ir,  198 Au,  199 Au,  211 Pb,  58 Co,  80m Br,  99m Tc,  103m Rh,  109 Pt,  119 Sb,  189m Os,  192 Ir,  219 Rn,  215 Po,  221 Fr,  217 At,  255 Fm,  11 C,  13 N,  15 O,  75 Br,  224 Ac,  126 I,  133 I,  77 Br,  113m In,  95 Ru,  97 Ru,  103 Ru,  105 Ru,  107 Hg,  203 Hg,  121m Te,  122m Te,  125m Te,  165 Tm,  167 Tm,  168 Tm,  197 Pt,  143 Pr,  57 Co,  51 Cr,  75 Se,  20 Tl,  76 Br and  169 Yb. 
     
     
         15 . The method of  claim 14 , wherein the radionuclide is  90 Y. 
     
     
         16 . The method of  claim 9 , wherein the radionuclide is an alpha-emitter. 
     
     
         17 . A method of delivering a diagnostic or therapeutic agent to a pancreatic cancer cell comprising:
 a) exposing the pancreatic cancer cell to a bispecific antibody comprising: (i) an anti-MUC-5ac antibody or antigen-binding fragment thereof that binds to the same epitope as or competes for binding to pancreatic cancer mucin with a chimeric PAM4 (cPAM4) antibody that comprises the light chain variable region CDR sequences CDR1 (SASSSVSSSYLY, (SEQ ID NO:1), CDR2 (STSNLAS, SEQ ID NO:2), and CDR3 (HQWNRYPYT, SEQ ID NO:3) and the heavy chain variable region CDR sequences CDR1 (SYVLH, SEQ ID NO:4), CDR2 (YINPYNDGTQYNEKFKG, SEQ ID NO:5) and CDR3 (GFGGSYGFAY, SEQ ID NO:6); and (ii) an anti-hapten antibody or antigen-binding fragment thereof; and   b) exposing the pancreatic cancer cell to a targetable construct attached to at least one therapeutic or diagnostic agent, wherein the targetable construct comprises at least one copy of the hapten.   
     
     
         18 . The method of  claim 17 , wherein the hapten is HSG (histamine-succinyl-glycine) or In-DTPA (diethylenetriaminepentaacetic acid). 
     
     
         19 . The method of  claim 17 , wherein the diagnostic agent is selected from the group consisting of a radionuclide, a contrast agent, a fluorescent agent, a chemiluminescent agent, a bioluminescent agent, a paramagnetic ion, an enzyme and a photoactive diagnostic agent. 
     
     
         20 . The method of  claim 19 , wherein the diagnostic agent is a radionuclide selected from the group consisting of  110 In,  111 In,  177 Lu,  18 F,  52 Fe,  62 Cu,  64 Cu,  67 Cu,  67 Ga,  68 Ga,  86 Y,  90 Y,  89 Zr,  94m Tc,  94 Tc,  99m Tc,  120 I,  123 I,  124 I,  125 I,  131 I,  154-158 Gd,  32 P,  11 C,  13 N,  15 O,  186 Re,  188 Re,  51 Mn,  52m Mn,  55 Co,  72 As,  75 Br,  76 Br,  82m Rb,  83 Sr, or other gamma-, beta-, or positron-emitters. 
     
     
         21 . The method of  claim 20 , wherein the radionuclide is  18 F. 
     
     
         22 . The method of  claim 19 , wherein the paramagnetic ion is selected from the group consisting of chromium (III), manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), gadolinium (III), vanadium (II), terbium (III), dysprosium (III), holmium (III) and erbium (III). 
     
     
         23 . The method of  claim 19 , wherein the diagnostic agent is a fluorescent labeling compound selected from the group consisting of fluorescein isothiocyanate, rhodamine, phycoerytherin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine, or a chemiluminescent labeling compound selected from the group consisting of luminol, isoluminol, an aromatic acridinium ester, an imidazole, an acridinium salt and an oxalate ester, or a bioluminescent compound selected from the group consisting of luciferin, luciferase and aequorin. 
     
     
         24 . The method of  claim 17 , wherein the therapeutic agent is selected from the group consisting of a radionuclide, an immunomodulator, a hormone, a hormone antagonist, an enzyme, an anti-sense oligonucleotide, an siRNA, an enzyme inhibitor, a photoactive therapeutic agent, a cytotoxic agent, a drug, a pro-drug, a toxin, an angiogenesis inhibitor and a pro-apoptotic agent. 
     
     
         25 . The method of  claim 24 , wherein the drug is selected from the group consisting of nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, gemcitabine, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzyme inhibitors, epipodophyllotoxins, platinum coordination complexes, vinca alkaloids, substituted ureas, methyl hydrazine derivatives, adrenocortical suppressants, hormone antagonists, endostatin, taxols, camptothecins, SN-38, doxorubicin, antimetabolites, alkylating agents, antimitotics, anti-angiogenic agents, tyrosine kinase inhibitors, mTOR inhibitors, heat shock protein (HSP90) inhibitors, proteosome inhibitors, HDAC inhibitors, pro-apoptotic agents, methotrexate and CPT-11. 
     
     
         26 . The method of  claim 24 , wherein the toxin is selected from the group consisting of ricin, abrin, alpha toxin, saporin, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin,  Pseudomonas  exotoxin, and  Pseudomonas  endotoxin. 
     
     
         27 . The method of  claim 24 , wherein the immunomodulator is selected from the group consisting of a cytokine, a stem cell growth factor, a lymphotoxin, a hematopoietic factor, a colony stimulating factor (CSF), an interleukin (IL), an interferon (IFN), a stem cell growth factor, erythropoietin, thrombopoietin, tumor necrosis factor (TNF), granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon-α, interferon-β, interferon-γ and the stem cell growth factor designated “S1 factor”. 
     
     
         28 . The method of  claim 27 , wherein the cytokine is selected from the group consisting of human growth hormone, N-methionyl human growth hormone, bovine growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, glycoprotein follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), placenta growth factor (PlGF), hepatic growth factor, prostaglandin, fibroblast growth factor, prolactin, placental lactogen, OB protein, tumor necrosis factor-α, tumor necrosis factor-β, mullerian-inhibiting substance, mouse gonadotropin-associated peptide, inhibin, activin, vascular endothelial growth factor, integrin, thrombopoietin (TPO), NGF-β, platelet-growth factor, TGF-α, TGF-β, insulin-like growth factor-I, insulin-like growth factor-II, erythropoietin (EPO), osteoinductive factors, interferon-α, interferon-β, interferon-γ, macrophage-CSF (M-CSF), IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, IL-25, LIF, FLT-3, angiostatin, thrombospondin, endostatin, TNF-α and LT (lymphotoxin). 
     
     
         29 . The method of  claim 24 , wherein the radionuclide is selected from the group consisting of  111 In,  177 Lu,  212 Bi,  213 Bi,  211 At,  62 Cu,  67 Cu,  90 Y,  125 I,  131 I,  32 P,  33 P,  47 Sc,  111 Ag,  67 Ga,  143 Pr,  153 Sm,  161 Tb,  166 Dy,  166 Ho,  186 Re,  188 Re,  189 Re,  212 Pb,  223 Ra,  225 Ac,  59 Fe,  75 Se,  77 As,  89 Sr,  99 Mo,  105 Rh,  109 Pd,  143 Pr,  149 Pm,  169 Er,  194 Ir,  198 Au,  199 Au,  211 Pb,  58 Co,  80m Br,  99m Tc,  103m Rh,  109 Pt,  119 Sb,  189m Os,  192 Ir,  219 Rn,  215 Po,  221 Fr,  217 At,  255 Fm,  11 C,  13 N,  15 O,  75 Br,  224 Ac,  126 I,  133 I,  77 Br,  113m In,  95 Ru,  97 Ru,  103 Ru,  105 Ru,  107 Hg,  203 Hg,  121m Te,  122m Te,  125m Te,  165 Tm,  167 Tm,  168 Tm,  197 Pt,  143 Pr,  57 Co,  51 Cr,  75 Se,  20 Tl,  76 Br and  169 Yb. 
     
     
         30 . The method of  claim 29 , wherein the radionuclide is  90 Y. 
     
     
         31 . The method of  claim 24 , wherein the radionuclide is an alpha-emitter.