Notch-based fusion proteins and uses thereof

Abstract

This invention provides a method for treating a subject having a tumor and a method for inhibiting angiogenesis in a subject, both comprising administering to the subject an effective amount of a composition of matter comprising the extracellular domain of a Notch receptor protein operably affixed to a half-life-increasing moiety. This invention also provides a composition of matter comprising the extracellular domain of Notch4 receptor protein operably affixed to a half-life-increasing moiety. This invention further provides an article of manufacture. Finally, this invention provides a replicable vector which encodes a polypeptide comprising the extracellular domain of a Notch receptor protein operably affixed to a half-life-increasing moiety, a host vector system which comprises such replicable vector and a method of producing such polypeptide.

1 - 90 . (canceled) 
     
     
         91 . A method for treating a subject having a tumor comprising administering to the subject an effective amount of a fusion protein comprising an extracellular domain of a human Notch receptor protein and an Fc portion of an antibody, wherein the extracellular domain of the Notch receptor protein comprises the EGF repeats of such Notch receptor protein and the extracellular domain is linked at its carboxyl terminus to the amino terminus of the Fc portion of an antibody, wherein the Notch receptor protein is a human Notch1, Notch2, Notch3, or Notch4 receptor protein, so as to thereby treat the subject. 
     
     
         92 . The method of  claim 91 , wherein the fusion protein comprises an extracellular domain of a human Notch receptor protein and an Fc portion of an antibody, wherein the extracellular domain of the Notch receptor protein comprises the EGF repeats of such Notch receptor protein and the extracellular domain is linked at its carboxyl terminus to the amino terminus of the Fc portion of an antibody by means of a peptide linker, wherein the Notch receptor protein is a human Notch1, Notch2, Notch3, or Notch4 receptor protein, the amino acid sequence of which is encoded by the nucleic acid sequence set forth in SEQ ID NOs. 11, 12, 13 and 14, respectively, and wherein the extracellular domain is bound to the Fc portion of the antibody by means of a peptide linker which comprises the consecutive amino acids DLGPG (SEQ ID NO:2). 
     
     
         93 . The method of  claim 91 , wherein the fusion protein consists of consecutive amino acids, the amino acid sequence of which, commencing at the amino terminus, is identical to the amino acid sequence of the EGF repeats of a human Notch receptor protein, followed by an amino acid sequence identical to the sequence of an Fc portion of an antibody, wherein the amino acid sequence of the EGF repeats of the human Notch receptor protein is selected from the group consisting of:
 (a) the amino acid sequence of EGF repeats 1-36 of human Notch1 encoded by the nucleic acid coding sequence of Notch1 EGF repeats 1-36 set forth in SEQ ID NO: 11;   (b) the amino acid sequence of EGF repeats 1-36 of human Notch2 encoded by the nucleic acid coding sequence of Notch2 EGF repeats 1-36 set forth in SEQ ID NO: 12;   (c) the amino acid sequence of EGF repeats 1-34 of human Notch3 encoded by the nucleic acid coding sequence of Notch3 EGF repeats 1-34 set forth in SEQ ID NO: 13; and   (d) the amino acid sequence of EGF repeats 1-29 of human Notch4 is encoded by the nucleic acid coding sequence of Notch4 EGF repeats 1-29 set forth in SEQ ID NO: 14.   
     
     
         94 . The method of  claim 91 , wherein the fusion protein consists of consecutive amino acids, the amino acid sequence of which, commencing at the amino terminus, is identical to the amino acid sequence of the EGF repeats of a human Notch receptor protein, followed by a linker sequence of amino acids which is followed by an amino acid sequence identical to the sequence of an Fc portion of an antibody, wherein the amino acid sequence of the EGF repeats of the human Notch receptor protein is selected from the group consisting of:
 (a) the amino acid sequence of EGF repeats 1-36 of human Notch1 encoded by the nucleic acid coding sequence of Notch1 EGF repeats 1-36 set forth in SEQ ID NO: 11;   (b) the amino acid sequence of EGF repeats 1-36 of human Notch2 encoded by the nucleic acid coding sequence of Notch2 EGF repeats 1-36 set forth in SEQ ID NO: 12;   (c) the amino acid sequence of EGF repeats 1-34 of human Notch3 encoded by the nucleic acid coding sequence of Notch3 EGF repeats 1-34 set forth in SEQ ID NO: 13; and   (d) the amino acid sequence of EGF repeats 1-29 of human Notch4 is encoded by the nucleic acid coding sequence of Notch4 EGF repeats 1-29 set forth in SEQ ID NO: 14.   
     
     
         95 . The method of  claim 94 , wherein the amino acid sequence of the EGF repeats of the human Notch receptor protein is identical to the amino acid sequence of EGF repeats 1-36 of human Notch1 encoded by the nucleic acid coding sequence of Notch1 EGF repeats 1-36 set forth in SEQ ID NO: 11. 
     
     
         96 . The method of  claim 94 , wherein the amino acid sequence of the EGF repeats of the human Notch receptor protein is identical to the amino acid sequence of EGF repeats 1-34 of human Notch3 encoded by the nucleic acid coding sequence of Notch3 EGF repeats 1-34 set forth in SEQ ID NO: 13. 
     
     
         97 . The method of  claim 94 , wherein the amino acid sequence of the EGF repeats of the human Notch receptor protein is identical to the amino acid sequence of EGF repeats 1-29 of human Notch4 is encoded by the nucleic acid coding sequence of Notch4 EGF repeats 1-29 set forth in SEQ ID NO: 14. 
     
     
         98 . The method of  claim 94 , wherein the Fc portion of the antibody is the Fc portion of a human antibody. 
     
     
         99 . A method of inhibiting angiogenesis in a subject comprising administering to the subject an effective amount of a fusion protein comprising an extracellular domain of a human Notch receptor protein and an Fc portion of an antibody, wherein the extracellular domain of the Notch receptor protein comprises the EGF repeats of such Notch receptor protein and the extracellular domain is linked at its carboxyl terminus to the amino terminus of the Fc portion of an antibody, wherein the Notch receptor protein is a human Notch1, Notch2, Notch3, or Notch4 receptor protein, so as to thereby treat the subject. 
     
     
         100 . The method of  claim 99 , wherein the fusion protein comprises an extracellular domain of a human Notch receptor protein and an Fc portion of an antibody, wherein the extracellular domain of the Notch receptor protein comprises the EGF repeats of such Notch receptor protein and the extracellular domain is linked at its carboxyl terminus to the amino terminus of the Fc portion of an antibody by means of a peptide linker, wherein the Notch receptor protein is a human Notch1, Notch2, Notch3, or Notch4 receptor protein, the amino acid sequence of which is encoded by the nucleic acid sequence set forth in SEQ ID NOs. 11, 12, 13 and 14, respectively, and wherein the extracellular domain is bound to the Fc portion of the antibody by means of a peptide linker which comprises the consecutive amino acids DLGPG (SEQ ID NO:2). 
     
     
         101 . The method of  claim 99 , wherein the fusion protein consists of consecutive amino acids, the amino acid sequence of which, commencing at the amino terminus, is identical to the amino acid sequence of the EGF repeats of a human Notch receptor protein, followed by an amino acid sequence identical to the sequence of an Fc portion of an antibody, wherein the amino acid sequence of the EGF repeats of the human Notch receptor protein is selected from the group consisting of:
 (a) the amino acid sequence of EGF repeats 1-36 of human Notch1 encoded by the nucleic acid coding sequence of Notch1 EGF repeats 1-36 set forth in SEQ ID NO: 11;   (b) the amino acid sequence of EGF repeats 1-36 of human Notch2 encoded by the nucleic acid coding sequence of Notch2 EGF repeats 1-36 set forth in SEQ ID NO: 12;   (c) the amino acid sequence of EGF repeats 1-34 of human Notch3 encoded by the nucleic acid coding sequence of Notch3 EGF repeats 1-34 set forth in SEQ ID NO: 13; and   (d) the amino acid sequence of EGF repeats 1-29 of human Notch4 is encoded by the nucleic acid coding sequence of Notch4 EGF repeats 1-29 set forth in SEQ ID NO: 14.   
     
     
         102 . The method of  claim 99 , wherein the fusion protein consists of consecutive amino acids, the amino acid sequence of which, commencing at the amino terminus, is identical to the amino acid sequence of the EGF repeats of a human Notch receptor protein, followed by a linker sequence of amino acids which is followed by an amino acid sequence identical to the sequence of an Fc portion of an antibody, wherein the amino acid sequence of the EGF repeats of the human Notch receptor protein is selected from the group consisting of:
 (a) the amino acid sequence of EGF repeats 1-36 of human Notch1 encoded by the nucleic acid coding sequence of Notch1 EGF repeats 1-36 set forth in SEQ ID NO: 11;   (b) the amino acid sequence of EGF repeats 1-36 of human Notch2 encoded by the nucleic acid coding sequence of Notch2 EGF repeats 1-36 set forth in SEQ ID NO: 12;   (c) the amino acid sequence of EGF repeats 1-34 of human Notch3 encoded by the nucleic acid coding sequence of Notch3 EGF repeats 1-34 set forth in SEQ ID NO: 13; and   (d) the amino acid sequence of EGF repeats 1-29 of human Notch4 is encoded by the nucleic acid coding sequence of Notch4 EGF repeats 1-29 set forth in SEQ ID NO: 14.   
     
     
         103 . The method of  claim 102 , wherein the amino acid sequence of the EGF repeats of the human Notch receptor protein is identical to the amino acid sequence of EGF repeats 1-29 of human Notch4 is encoded by the nucleic acid coding sequence of Notch4 EGF repeats 1-29 set forth in SEQ ID NO: 14. 
     
     
         104 . The method of  claim 102 , wherein the Fc portion of the antibody is the Fc portion of a human antibody. 
     
     
         105 . The method of  claim 99 , wherein the angiogenesis is tumor angiogenesis. 
     
     
         106 . The method of  claim 99 , wherein the subject is afflicted with a pathologic vascular hyperplasia or lymphatic vascular proliferative disease.