Monoclonal Antibody hPAM4

Abstract

This invention relates to monovalent and multivalent, monospecific antibodies and to multivalent, multispecific antibodies. One embodiment of these antibodies has one or more identical binding sites where each binding site binds with a target antigen or an epitope on a target antigen. Another embodiment of these antibodies has two or more binding sites where these binding sites have affinity towards different epitopes on a target antigen or different target antigens, or have affinity towards a target antigen and a hapten. The present invention further relates to recombinant vectors useful for the expression of these functional antibodies in a host. More specifically, the present invention relates to the tumor-associated antibody designated PAM4. The invention further relates to humanized and human PAM4 antibodies, and the use of such antibodies in diagnosis and therapy.

1 . A method of delivering a diagnostic or therapeutic agent, or a combination thereof, to a target comprising 
 (a) providing a composition that comprises a humanized PAM4 (hPAM4) antibody or antigen-binding fragment thereof conjugated to at least one diagnostic and/or therapeutic agent, wherein said hPAM4 antibody or fragment binds to a domain located between the amino terminus and the start of the repeat domain of a MUC1 antigen and does not bind to the repeat domain of MUC1; and    (b) administering the composition to a subject.    
     
     
         2 . The method of  claim 1 , wherein said diagnostic agent is selected from the group consisting of a radionuclide, a contrast agent, and a photoactive diagnostic agent.  
     
     
         3 . The method of  claim 2 , wherein said diagnostic agent is a radionuclide.  
     
     
         4 . The method of  claim 3 , wherein said radionuclide has an energy between 20 and 4,000 keV.  
     
     
         5 . The method of  claim 4 , wherein said radionuclide is a gamma-, beta- or a positron-emitting isotope.  
     
     
         6 . The method of  claim 5 , wherein said radionuclide is selected from the group consisting of 110In, 111In, 177Lu, 18F, 52Fe, 62Cu, 64Cu, 67Cu, 67Ga, 2Ga, 16Y, 20Y, 89Zr, 94mTc, 94Tc, 99mTc 120I, 123I, 124I, 125I, 131I, 154-158Gd, 32P, 11C, 13N, 15O, 116Re 188Re, 51Mn, 52mMn, 55Co, 72As, 75Br, 76Br, 82mRb, 83Sr, or other gamma-, beta-, or positron-emitters.  
     
     
         7 . The method of  claim 2 , wherein said diagnostic agent is a contrast agent.  
     
     
         8 . The method of  claim 7 , wherein said contrast agent is a paramagnetic ion.  
     
     
         9 . The method of  claim 8 , wherein said paramagnetic ion is a metal comprising 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) or erbium (III).  
     
     
         10 . The method of  claim 7 , wherein said contrast agent is a metal comprising lanthanum (III), gold (III), lead (II) or bismuth (III).  
     
     
         11 . The method of  claim 7 , wherein said contrast agent is an ultrasound enhancing agent.  
     
     
         12 . The method of  claim 11 , wherein said ultrasound enhancing agent is a liposome.  
     
     
         13 . The method of  claim 7 , wherein said contrast agent is a radiopaque material selected from the group comprising iodine compounds, barium compounds, gallium compounds, and thallium compounds.  
     
     
         14 . The method of  claim 13 , wherein said radiopaque material is selected from the group comprising barium, diatrizoate, ethiodized oil, gallium citrate, iocarmic acid, iocetamic acid, iodamide, iodipamide, iodoxamic acid, iogulamide, iohexyl, iopamidol, iopanoic acid, ioprocemic acid, iosefamic acid, ioseric acid, iosulamide meglumine, iosemetic acid, iotasul, iotetric acid, iothalamic acid, iotroxic acid, ioxaglic acid, ioxotrizoic acid, ipodate, meglumine, metrizamide, metrizoate, propyliodone, and thallous chloride.  
     
     
         15 . The method of  claim 2 , wherein said diagnostic agent is a photoactive diagnostic agent.  
     
     
         16 . The method of  claim 1 , wherein said therapeutic agent is selected from the group consisting of a cytotoxic agent, cytokine, immunomodulator, hormone, a hormone antagonist, growth factor, radionuclide, metal, contrast agent, oligonucleotide, enzyme, enzyme inhibitor, and photoactive therapeutic agent.  
     
     
         17 . The therapeutic conjugate of  claim 16 , wherein said oligonucleotide is an antisense oligonucleotide.  
     
     
         18 . The oligonucleotide of  claim 17 , wherein said antisense oligonucleotide is an antisense oligonucleotide against an oncogene.  
     
     
         19 . The antisense oligonucleotide of  claim 18 , wherein said oncogene is bcl-2 or p53.  
     
     
         20 . The method of  claim 16 , wherein said therapeutic agent is a cytotoxic agent.  
     
     
         21 . The method of  claim 20 , wherein said cytotoxic agent is a drug or a toxin.  
     
     
         22 . The method of  claim 21 , wherein said drug possesses the pharmaceutical property selected from the group consisting of antimitotic, alkylating, antimetabolite, antiangiogenic, apoptotic, alkaloid, and antibiotic agents and combinations thereof.  
     
     
         23 . The method of  claim 21 , wherein said drug is selected from the group consisting of nitrogen mustards, gemcitabine, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, triazenes, folic acid analogs, anthracyclines, SN-38, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzymes, enzyme inhibitors, epipodophyllotoxins, platinum coordination complexes, vinca alkaloids, substituted ureas, methyl hydrazine derivatives, adrenocortical suppressants, hormone antagonists, endostatin, taxols, camptothecins, doxorubicins and their analogs, antimetabolites, alkylating agents, antimitotics, antiangiogenic, apoptotic agents, methotrexate, CPT-11, and a combination thereof.  
     
     
         24 . The method of  claim 21 , wherein said toxin is selected from the group consisting of ricin, abrin, alpha toxin, saporin, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtherin toxin,  Pseudomonas  exotoxin, and  Pseudomonas  endotoxin.  
     
     
         25 . The method of  claim 16 , wherein said therapeutic agent is an immunomodulator.  
     
     
         26 . The method of  claim 25 , wherein said 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 interferon (IFN), a stem cell growth factor, erythropoietin, thrombopoietin and a combination thereof.  
     
     
         27 . The method of  claim 26 , wherein said lymphotoxin is tumor necrosis factor (TNF), said hematopoietic factor is an interleukin (IL), said colony stimulating factor is granulocyte-colony stimulating factor (G-CSF) or granulocyte macrophage-colony stimulating factor (GM-CSF)), said interferon is interferons-alpha, -beta or -gamma, and said stem cell growth factor is designated “SI factor”.  
     
     
         28 . The method of  claim 25 , wherein said immunomodulator is a cytokine.  
     
     
         29 . The method of  claim 25 , wherein said immunomodulator comprises IL-1, IL-2, IL-3, IL-6, IL-10, IL-12, IL-18, IL-21, interferon-alpha, beta, or gamma, TNF-alpha or a combination thereof.  
     
     
         30 . The method of  claim 16 , wherein said therapeutic agent is a radionuclide.  
     
     
         31 . The method of  claim 30 , wherein said radionuclide has an energy between 60 and 700 keV.  
     
     
         32 . The method of  claim 31 , wherein said radionuclide is selected from the group consisting of 32P, 33P, 47Sc, 64Cu, 67Cu, 67Ga, 16Y, 20Y, 111Ag, 111In, 125I, 131I, 142Pr, 153Sm, 161Tb, 166Dy, 166Ho, 177Lu, 116Re, 188Re, 189Re, 212Pb, 212Bi, 213Bi, 211At, 223Ra and 225Ac, and combinations thereof.  
     
     
         33 . The method of  claim 16 , wherein said therapeutic agent is a photoactive therapeutic agent.  
     
     
         34 . The method of  claim 16 , wherein said therapeutic agent is an enzyme.  
     
     
         35 . The method of  claim 1 , wherein the hPAM4 antibody or fragment comprises the complementarity-determining regions (CDRs) of a murine PAM4 monoclonal antibody (MAb) and the framework (FR) regions of the light and heavy chain variable regions of a human antibody and the light and heavy chain constant regions of a human antibody, wherein the CDRs of the light chain variable region of the humanized PAM4 Mab comprise CDR1 (SASSSVSSSYLY, SEQ ID NO:1); CDR2 (STSNLAS, SEQ ID NO:2); and CDR3 (HQWNRYPYT, SEQ ID NO:3); and the CDRs of the heavy chain variable region of the humanized PAM4 Mab comprise CDR1 (SYVLH, SEQ ID NO:4); CDR2 (YINPYNDGTQYNEKFKG, SEQ ID NO:5) and CDR3 (GFGGSYGFAY, SEQ ID NO:6).  
     
     
         36 . A method of delivering a diagnostic agent, a therapeutic agent, or a combination thereof to a target, comprising: 
 (a) administering to a subject a composition comprising a first hPAM4 antibody or fragment thereof that binds to a domain located between the amino terminus and the start of the repeat domain of a MUC1 antigen and does not bind to the repeat domain of MUC1, and a second antibody or fragment thereof that binds to a carrier molecule;    (b) waiting a sufficient amount of time for an amount of the composition to clear the subject's blood stream; and    (c) administering to said subject a carrier molecule comprising a diagnostic agent, a therapeutic agent, or a combination thereof.    
     
     
         37 . The method of  claim 36 , wherein said diagnostic agent or said therapeutic agent is selected from the group comprising isotopes, drugs, toxins, cytokines, hormones, hormone antagonists, oligonucleotides, enzymes, enzyme inhibitors, growth factors, radionuclides, and metals.  
     
     
         38 . The method of  claim 36 , wherein the hPAM4 antibody or fragment comprises the complementarity-determining regions (CDRs) of a murine PAM4 monoclonal antibody (MAb) and the framework (FR) regions of the light and heavy chain variable regions of a human antibody and the light and heavy chain constant regions of a human antibody, wherein the CDRs of the light chain variable region of the humanized PAM4 Mab comprise CDR1 (SASSSVSSSYLY, SEQ ID NO:1); CDR2 (STSNLAS, SEQ ID NO:2); and CDR3 (HQWNRYPYT, SEQ ID NO:3); and the CDRs of the heavy chain variable region of the humanized PAM4 Mab comprise CDR1 (SYVLH, SEQ ID NO:4); CDR2 (YINPYNDGTQYNEKFKG, SEQ ID NO:5) and CDR3 (GFGGSYGFAY, SEQ ID NO:6).  
     
     
         39 . A method for diagnosing or treating cancer, comprising: 
 (a) administering to a subject a composition comprising a first hPAM4 antibody or fragment thereof that binds to a domain located between the amino terminus and the start of the repeat domain of a MUC1 antigen and does not bind to the repeat domain of MUC1, and a second antibody or fragment thereof that binds to a carrier molecule;    (b) waiting a sufficient amount of time for an amount of the antibody to clear the subject's blood stream; and    (c) administering to said subject a carrier molecule comprising a diagnostic agent, a therapeutic agent, or a combination thereof.    
     
     
         40 . The method of  claim 39 , wherein said cancer is pancreatic cancer.  
     
     
         41 . The method of  claim 39 , wherein said method is used for intraoperative identification of diseased tissues.  
     
     
         42 . The method of  claim 39 , wherein said method is used for endoscopic identification of diseased tissues.  
     
     
         43 . The method of  claim 39 , wherein said method is used for intravascular identification of diseased tissues.  
     
     
         44 . The method of  claim 39 , wherein the hPAM4 antibody or fragment comprises the complementarity-determining regions (CDRs) of a murine PAM4 monoclonal antibody (MAb) and the framework (FR) regions of the light and heavy chain variable regions of a human antibody and the light and heavy chain constant regions of a human antibody, wherein the CDRs of the light chain variable region of the humanized PAM4 Mab comprise CDR1 (SASSSVSSSYLY, SEQ ID NO:1); CDR2 (STSNLAS, SEQ ID NO:2); and CDR3 (HQWNRYPYT, SEQ ID NO:3); and the CDRs of the heavy chain variable region of the humanized PAM4 Mab comprise CDR1 (SYVLH, SEQ ID NO:4); CDR2 (YINPYNDGTQYNEKFKG, SEQ ID NO:5) and CDR3 (GFGGSYGFAY, SEQ ID NO:6).