US2003032609A1PendingUtilityA1

Methods of modulating of angiogenesis

Priority: Aug 14, 1998Filed: Apr 12, 2002Published: Feb 13, 2003
Est. expiryAug 14, 2018(expired)· nominal 20-yr term from priority
A61K 38/00G06F 16/289C07K 14/47
52
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Claims

Abstract

A method of inhibiting angiogenesis in a mammal by administering to the mammal a compound which inhibits binding of endothelial PAS domain protein-1 to cis-acting transcription regulatory sequence in the promoter region of a gene encoding an angiogenic factor.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A method of inhibiting angiogenesis in a mammal comprising administering to said mammal a compound which inhibits binding of endothelial PAS domain protein-1 (EPAS1) to cis-acting transcription regulatory DNA of an angiogenic factor.  
     
     
         2 . The method of  claim 1 , wherein said angiogenic factor is a vascular endothelial growth factor receptor (VEGF-R).  
     
     
         3 . The method of  claim 2 , wherein said receptor is KDR/flk-1.  
     
     
         4 . The method of  claim 2 , wherein said receptor is flt-1.  
     
     
         5 . The method of  claim 1 , wherein said angiogenic factor is vascular endothelial growth factor (VEGF).  
     
     
         6 . The method of  claim 1 , wherein said angiogenic factor is Tie2.  
     
     
         7 . The method of  claim 1 , wherein said compound inhibits transcription of said angiogenic factor.  
     
     
         8 . The method of  claim 1 , wherein said regulatory DNA comprises GCCCTACGTGCTGTCTCA (SEQ ID NO: 1 ).  
     
     
         9 . The method of  claim 1 , wherein said compound is an EPAS1 polypeptide lacking a transactivation domain.  
     
     
         10 . The method of  claim 9 , wherein said transactivation domain comprises the amino acid sequence of SEQ ID NO:  2 .  
     
     
         11 . The method of  claim 9 , wherein said transactivation domain comprises the amino acids  486 - 639  of SEQ ID NO:  6 .  
     
     
         12 . The method of  claim 9 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:  4 .  
     
     
         13 . The method of  claim 1 , wherein said compound is a nucleic acid encoding an EPAS1 polypeptide lacking the amino acid sequence of SEQ ID NO:  2 .  
     
     
         14 . The method of  claim 1 , wherein said compound is a nucleic acid encoding an EPAS1 polypeptide lacking amino acids  486 - 639  of SEQ ID NO:  6 .  
     
     
         15 . The method of  claim 1 , wherein said compound is a antisense nucleic acid molecule comprising at least 10 nucleotides, wherein the sequence of said molecule is complementary to part of or all ofan MRNA encoding EPAS1 polypeptide.  
     
     
         16 . The method of  claim 1 , wherein said compound is an EPAS1-specific intrabody.  
     
     
         17 . The method of  claim 1 , wherein said compound is administered to a site of an atherosclerotic lesion in said mammal.  
     
     
         18 . The method of  claim 1 , wherein said compound is administered to a tumor site in said mammal.  
     
     
         19 . An antibody which binds to EPAS1 .  
     
     
         20 . The antibody of  claim 19 , wherein said antibody binds to a C-terminal activation domain of EPAS1.  
     
     
         21 . The antibody of  claim 20 , wherein said activation domain comprises SEQ ID NO:  2 .  
     
     
         22 . A method of promoting angiogenesis in a mammal comprising administering to said mammal a compound which increases expression of VEGF or a VEGF-R in an endothelial cell.  
     
     
         23 . The method of  claim 19 , wherein said VEGF-R is KDR/flk-1 or flt-1.  
     
     
         24 . A substantially pure DNA comprising a sequence encoding a aryl hydrocarbon receptor nuclear translocator-4 (ARNT4) polypeptide.  
     
     
         25 . The DNA of  claim 24 , wherein said DNA encodes a human ARNT4 polypeptide.  
     
     
         26 . The DNA of  claim 24 , wherein said polypeptide comprises the amino acid sequence of residues  75  to  128 , inclusive, of SEQ ID NO:  19 .  
     
     
         27 . The DNA of  claim 24 , wherein said polypeptide comprises the amino acid sequence of residues  155  to  207 , inclusive, of SEQ ID NO:  19 .  
     
     
         28 . The DNA of  claim 24 , wherein said polypeptide comprises the amino acid sequence of residues  232  to  384 , inclusive, of SEQ ID NO:  19 .  
     
     
         29 . A substantially pure DNA comprising a nucleotide sequence having at least 50% sequence identity to SEQ ID NO:  20 .  
     
     
         30 . The DNA of  claim 24 , wherein said DNA comprises the coding sequences of SEQ ID NO:  20 .  
     
     
         31 . The DNA of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:  19 .  
     
     
         32 . A substantially pure DNA comprising a strand which hybridizes at high stringency to a strand of DNA consisting of the coding sequence of SEQ ID NO:  20 , or the complement thereof.  
     
     
         33 . A substantially pure DNA comprising a sequences at least 50% sequence identity to the coding sequence of SEQ ID NO:  20 , and encoding a polypeptide having the biological activity of an ARNT4 polypeptide.  
     
     
         34 . A substantially pure ARNT4 polypeptide.  
     
     
         35 . The polypeptide of  claim 34 , wherein said polypeptide is human ARNT4.  
     
     
         36 . The polypeptide of  claim 34 , wherein said polypeptide comprises the amino acid sequence of residues  75  to  128 , inclusive, of SEQ ID NO:  19 .  
     
     
         37 . The polypeptide of  claim 34 , wherein said polypeptide comprises the amino acid sequence of residues  155  to  207 , inclusive, of SEQ ID NO:  19 .  
     
     
         38 . The polypeptide of  claim 34 , wherein said polypeptide comprises the amino acid sequence of residues  232  to  384 , inclusive, of SEQ ID NO:  19 .  
     
     
         39 . The polypeptide of  claim 34 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:  19 .  
     
     
         40 . The polypeptide of  claim 34 , wherein said polypeptide comprises an amino acid sequence at least 50% identical to SEQ ID NO:  19 .  
     
     
         41 . The polypeptide of  claim 34 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:  19 .  
     
     
         42 . A vector comprising the DNA of  claim 24 .  
     
     
         43 . A host cell comprising the DNA of  claim 24 .  
     
     
         44 . A transgenic non-human animal the germ cells and nucleated somatic cells of which comprise a null mutation in a gene encoding ARNT4.  
     
     
         45 . A method of inhibiting angiogenesis in a mammal comprising administering to said mammal a compound which inhibits binding of EPAS1 to ARNT4.  
     
     
         46 . The method of  claim 45 , wherein said compound is an ARNT4 polypeptide.  
     
     
         47 . The method of  claim 45 , wherein said polypeptide comprises the amino acid sequence of residues  75  to  128 , inclusive, of SEQ ID NO:  19 .  
     
     
         48 . The method of  claim 45 , wherein said polypeptide comprises the amino acid sequence of residues  155  to  207 , inclusive, of SEQ ID NO:  19 .  
     
     
         49 . The method of  claim 45 , wherein said polypeptide comprises the amino acid sequence of residues  232  to  384 , inclusive, of SEQ ID NO:  19 .  
     
     
         50 . The method of  claim 45 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:  19 .  
     
     
         51 . An EPAS1 polypeptide lacking a transactivation domain.  
     
     
         52 . The polypeptide of  claim 51 , wherein said transactivation domain comprises the amino acid sequence of SEQ ID NO:  2 .  
     
     
         53 . The polypeptide of  claim 51 , wherein said transactivation domain comprises the amino acids  486 - 639  of SEQ ID NO:  6 .  
     
     
         54 . The polypeptide of  claim 51 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:  4 .  
     
     
         55 . A nucleic acid encoding an EPAS1 polypeptide lacking the amino acid sequence of SEQ ID NO:  2 .  
     
     
         56 . The nucleic acid of  claim 55 , wherein said nucleic acid encodes an EPASI polypeptide lacking amino acids  486 - 639  of SEQ ID NO:  6 .

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