Her-2 binding antagonists

Abstract

There is disclosed a pharmaceutical composition for treating solid tumors that overexpress HER-2, comprising an agent selected from the group consisting of (a) an isolated polypeptide having from about 50 to 79 amino acids taken from the sequence of SEQ ID NO. 1 or SEQ ID NO:12, wherein the polypeptide binds to the extracellular domain ECD of HER-2 at an affinity of at least 10 8 , (b) an isolated and glycosylated polypeptide having from about 300 to 419 amino acids taken from the sequence of SEQ ID NO:2 or SEQ ID NO:13, wherein the C terminal 79 amino acids are present, and wherein at least three N-linked glycosylation sites are present, (c) a monoclonal antibody that binds to the ECD of HER-2, and (d) combinations thereof, with the proviso that the agent cannot be the monoclonal antibody alone, and pharmaceutically acceptable carrier. Also disclosed are prognostic and diagnostic assays.

1 .- 23 . (canceled) 
     
     
         24 . A method for determining the prognosis of tumor treatment in a patient for a tumor that overexpresses HER-2, comprising: (a) obtaining a bodily fluid sample from a patient, wherein the bodily fluid is selected from the group consisting blood, serum, urine, lymph, saliva, tumor tissue, placental tissue, umbilical cord tissue, amniotic fluid, chorionic villi tissue and combinations thereof; and (b) measuring the amount of p68HER-2 expressed using an anti-p68HER-2 antibody-based assay, wherein the assay is selected from the group consisting of ELISA, immunoprecipitation, immunohistocytochemistry, and Western analysis. 
     
     
         25 . The method for determining the prognosis of tumor treatment for a tumor that overexpresses HER-2 of  claim 24 , further comprising measuring the amount of p185HER-2 ECD in the bodily fluid. 
     
     
         26 . The method for determining the prognosis of tumor treatment for a tumor that overexpresses HER-2 of  claim 25 , further comprising determining a ratio between the amount of p68HER-2 and p185HER-2, whereby the higher the p68HER-2 to p185HER-2 ratio, the better the prognosis of the patient. 
     
     
         27 . An assay for cancer treatment, prognosis or diagnosis in a patient comprising:
 (a) obtaining a bodily fluid sample from the patient, wherein the bodily fluid is selected from the group consisting of blood, serum, urine, lymph, saliva, tumor tissue, placental tissue, umbilical cord tissue, amniotic fluid, chorionic villi tissue and combinations thereof;   (b) determining whether an ECDIIIa variant protein or DNA sequence, or a HER-2 intron 8 variant DNA sequence is present in the bodily fluid sample using a sequence identity assay; and   (c) correlating the presence of the ECDIIIa variant protein or DNA sequence, or the HER-2 intron 8 variant DNA sequence to cancer treatment and diagnosis using an historical database.   
     
     
         28 . The diagnostic assay of  claim 27 , wherein the sequence identity assay is selected from the group consisting of DNA sequencing, PCR assays, ELISA immunologic assays, immunoassays, hybridization assays, and combinations thereof. 
     
     
         29 . The diagnostic assay of  claim 27 , further comprising measuring the amount of p185HER-2 ECD in the bodily fluid. 
     
     
         30 . An assay for cancer treatment, prognosis or diagnosis in a patient comprising:
 (a) obtaining a bodily fluid sample from the patient, wherein the bodily fluid is selected from the group consisting of blood, serum, urine, lymph, saliva, tumor tissue, placental tissue, umbilical cord tissue, amniotic fluid, chorionic villi tissue and combinations thereof;   (b) determining whether a HER-2 intron 8 variant DNA sequence is present in the bodily fluid sample using a sequence identity assay; and   (c) correlating the presence of the HER-2 intron 8 variant DNA sequence to cancer treatment and diagnosis using an historical database.   
     
     
         31 . The diagnostic assay of  claim 30 , wherein the sequence identity assay is selected from the group consisting of DNA sequencing, PCR assays, hybridization assays, and combinations thereof. 
     
     
         32 . The diagnostic assay of  claim 30 , further comprising measuring the amount of p185HER-2 ECD in the bodily fluid. 
     
     
         33 . An assay for cancer treatment, prognosis or diagnosis in a patient comprising:
 (a) obtaining a bodily fluid sample from the patient, wherein the bodily fluid is selected from the group consisting of blood, serum, urine, lymph, saliva, tumor tissue, placental tissue, umbilical cord tissue, amniotic fluid, chorionic villi tissue and combinations thereof;   (b) determining whether an amount of an p68HER-2 ECDIIIa variant is present in the bodily fluid sample using an anti-p68HER-2 antibody-based assay, wherein the assay is selected from the group consisting of ELISA, immunoprecipitation, immunohistocytochemistry, and Western analysis; and   (c) correlating the presence or amount of the p68HER-2 ECDIIIa variant to cancer treatment and diagnosis using an historical database.   
     
     
         34 . The diagnostic assay of  claim 33 , wherein the sequence identity assay is selected from the group consisting of DNA sequencing, PCR assays, ELISA immunologic assays, hybridization assays, and combinations thereof. 
     
     
         35 . The diagnostic assay of  claim 33 , further comprising measuring the amount of p185HER-2 ECD in the bodily fluid. 
     
     
         36 . The diagnostic assay of  claim 35 , further comprising determining a ratio between the amount of p68HER-2 and p185HER-2 ECD. 
     
     
         37 . An antibody specific for an ECDIIIa variant of the sequence in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:12 or SEQ ID NO:13. 
     
     
         38 . An antibody specific for p68HER-2 ECDIIIa variant 3. 
     
     
         39 . A diagnostic kit comprising:
 (a) a monoclonal antibody or antigen-binding fragment thereof that specifically binds to an ECDIIIa variant of the sequence in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:12 or SEQ ID NO:13; and   (b) a detectable label, whereby the binding of the antibody in step (a) can be detected.   
     
     
         40 . The diagnostic kit of  claim 36 , wherein the label is selected from the group consisting of enzymes, radiolabels, chromohphores, chemiluminescent tags, and fluorescers.