Adenovirus-mediated intratumoral delivery of an angiogenesis antagonist for the treatment of tumors

Abstract

The present invention relates to gene therapy for the treatment of tumors. The invention more particularly relates to introduction of a gene encoding an anti-angiogenic factor into cells of a tumor, for example with a defective adenovirus vector, to inhibit growth or metastasis, or both, of the tumor. In a specific embodiment, delivery of a defective adenovirus that expresses the amino terminal fragment of urokinase (ATF) inhibited growth and metastasis of tumors. These effects were correlated with a remarkable inhibition of neovascularization within, and at the immediate vicinity of, the injection site. Delivery of a defective adenovirus vector that expresses kringles 1 to 3 of angiostatin inhibited tumor growth and tumorigenicity, and induced apoptosis of tumor cells. The invention further provides viral vectors for use in the methods of the invention.

What is claimed is:  
     
         1 . A method for inhibiting growth of a tumor comprising introducing into the tumor a defective adenovirus vector comprising a gene encoding an anti-angiogenic factor operably associated with an expression control sequence that provides for expression of the anti-angiogenic factor in a cell of the tumor.  
     
     
         2 . The method according to  claim 1 , wherein the tumor is a lung carcinoma or a breast carcinoma.  
     
     
         3 . The method according to  claim 1 , wherein the anti-angiogenic factor comprises a sequence of an amino terminal fragment of urokinase having an EGF-like domain, with the proviso that the factor is not urokinase.  
     
     
         4 . The method according to  claim 3 , wherein the anti-angiogenic factor is an amino terminal fragment of urokinase having an amino acid sequence of urokinase from about amino acid residue 1 to about residue 135.  
     
     
         5 . The method according to  claim 4 , wherein the urokinase is murine urokinase.  
     
     
         6 . The method according to  claim 4 , wherein the urokinase is human urokinase.  
     
     
         7 . The method according to  claim 1 , wherein the anti-angiogenic factor is angiostatin.  
     
     
         8 . The method according to  claim 7 , wherein the angiostatin comprises kringles 1 to 3.  
     
     
         9 . The method according to  claim 7 , wherein the angiostatin is an amino terminal fragment of plasminogen (Plg) having an amino acid sequence of plasminogen from about amino acid residue 1 to about residue 333.  
     
     
         10 . The method according to  claim 9 , wherein the plasminogen is human plasminogen.  
     
     
         11 . A method for inhibiting growth or metastasis, or both, of a tumor comprising introducing a vector comprising a gene encoding an amino terminal fragment of urokinase having an EGF-like domain into the tumor, with the proviso that the gene does not encode urokinase, wherein the gene is operably associated with an expression control sequence that provides for expression of the gene in a cell of the tumor.  
     
     
         12 . The method according to  claim 11 , wherein the amino terminal fragment of urokinase has an amino acid sequence of urokinase from about amino acid residue 1 to about residue 135.  
     
     
         13 . The method according to  claim 12 , wherein the urokinase in murine urokinase.  
     
     
         14 . The method according to  claim 12 , wherein the urokinase in human urokinase.  
     
     
         15 . A defective adenovirus vector comprising a gene encoding an anti-angiogenic factor operably associated with an expression control sequence.  
     
     
         16 . The virus vector according to  claim 15 , wherein the anti-angiogenic factor comprises a nucleic acid sequence of an amino terminal fragment of urokinase having an EGF-like domain, with the proviso that the factor is not urokinase.  
     
     
         17 . A defective adenovirus vector comprising a gene encoding an amino terminal fragment of urokinase having an EGF-like domain, with the proviso that the gene does not encode urokinase.  
     
     
         18 . The virus vector according to  claim 17 , wherein the amino terminal fragment of urokinase has an amino acid sequence of urokinase from amino acid residue 1 to about residue 135.  
     
     
         19 . The virus vector according to  claim 18 , wherein the urokinase is murine urokinase.  
     
     
         20 . The virus vector according to  claim 18 , wherein the urokinase is human urokinase.  
     
     
         21 . The virus vector according to  claim 15 , wherein the anti-angiogenic factor is angiostatin.  
     
     
         22 . The virus vector according to  claim 21 , wherein the angiostatin comprises kringles 1 to 3.  
     
     
         23 . The virus vector according to  claim 21 , wherein the angiostatin comprises a nucleic acid sequence of an amino terminal fragment of plasminogen having an amino acid sequence of plasminogen from amino acid residue 1 to about residue 333.  
     
     
         24 . The virus vector according to  claim 23 , wherein the plasminogen is human plasminogen.  
     
     
         25 . A pharmaceutical composition comprising a virus vector of any one of claims  15 - 24  and a pharmaceutically acceptable carrier.  
     
     
         26 . Use of the virus vector of any one of claims  15 - 24  in the manufacture of a medicament for inhibiting growth of a tumor.  
     
     
         27 . Use of the virus vector of any one of claims  16 - 20  in the manufacture of a medicament for inhibiting growth, or metastasis, or both of a tumor.  
     
     
         28 . Use of the virus vector of any one of claims  21 - 24  in the manufacture of a medicament for inhibiting tumor growth and inducing apoptosis.  
     
     
         29 . Use of a vector comprising a gene encoding an amino-terminal fragment of urokinase having an EGF-like domain, with the proviso that the gene does not encode urokinase, operably associated with an expression control sequence that provides for expression of the anti-angiogenic factor in the manufacture of a medicament for inhibiting growth or metastasis, or both, of a tumor.  
     
     
         30 . The use according to any of claims  26 - 29 , wherein the tumor is a lung carcinoma or a breast carcinoma.  
     
     
         33 . The method according to claim  31 , wherein the anti-angiogenic factor comprises an amino terminal fragment of urokinase comprising an EGF-like domain, with the exception that the anti-angiogenic factor is not urokinase.  
     
     
         34 . The method according to  claim 33 , wherein the anti-angiogenic factor is an amino terminal fragment of urokinase comprising an amino acid sequence of urokinase from about amino acid residue 1 to about residue 135.  
     
     
         35 . The method according to  claim 34 , wherein the urokinase is murine urokinase.  
     
     
         36 . The method according to  claim 34 , wherein the urokinase is human urokinase.  
     
     
         37 . The method according to claim  31 , wherein the anti-angiogenic factor is angiostatin.  
     
     
         38 . The method according to  claim 37 , wherein the angiostatin comprises kringles 1 to 3.  
     
     
         39 . The method according to  claim 37 , wherein the angiostatin is an amino terminal fragment of plasminogen (Plg) comprising an amino acid sequence of plasminogen from about amino acid residue 1 to about residue 333.  
     
     
         40 . The method according to  claim 39 , wherein the plasminogen is human plasminogen.  
     
     
         41 . A method for inhibiting growth and/or metastasis of a tumor comprising introducing a vector comprising a gene encoding an amino terminal fragment of urokinase comprising an EGF-like domain into the tumor, with the exception that the gene does not encode urokinase, wherein the gene is operably associated with an expression control sequence that provides for expression of the gene in a cell of the tumor.  
     
     
         42 . The method according to  claim 41 , wherein the amino terminal fragment of urokinase comprises an amino acid sequence of urokinase from about amino acid residue 1 to about residue 135.  
     
     
         43 . The method according to  claim 42 , wherein the urokinase is murine urokinase.  
     
     
         44 . The method according to  claim 42 , wherein the urokinase is human urokinase.  
     
     
         45 . A defective adenovirus vector comprising a gene encoding an anti-angiogenic factor operably associated with an expression control sequence.  
     
     
         46 . The defective adenovirus vector according to  claim 45 , wherein the anti-angiogenic factor comprises an amino terminal fragment of urokinase comprising an EGF-like domain, with the exception that the anti-angiogenic factor is not urokinase.  
     
     
         47 . A defective adenovirus vector comprising a gene encoding an amino terminal fragment of urokinase comprising an EGF-like domain, with the exception that the gene does not encode urokinase.  
     
     
         48 . The defective adenovirus vector according to  claim 47 , wherein the amino terminal fragment of urokinase comprises an amino acid sequence of urokinase from about amino acid residue 1 to about residue 135.  
     
     
         49 . The defective adenovirus vector according to  claim 48 , wherein the urokinase is murine urokinase.  
     
     
         50 . The defective adenovirus vector according to  claim 48 , wherein the urokinase is human urokinase.  
     
     
         51 . The defective adenovirus vector according to  claim 45 , wherein the anti-angiogenic factor is angiostatin.  
     
     
         52 . The defective adenovirus vector according to  claim 51 , wherein the angiostatin comprises kringles 1 to 3.  
     
     
         53 . The defective adenovirus vector according to  claim 51 , wherein the angiostatin comprises an amino terminal fragment of plasminogen comprising an amino acid sequence of plasminogen from about amino acid residue 1 to about residue 333.  
     
     
         54 . The defective adenovirus vector according to  claim 53 , wherein the plasminogen is human plasminogen.  
     
     
         55 . A pharmaceutical composition comprising the defective adenovirus vector according to  claim 45  and a pharmaceutically acceptable carrier.  
     
     
         56 . A pharmaceutical composition comprising the defective adenovirus vector according to  claim 47  and a pharmaceutically acceptable carrier.