Replication deficient adenoviral TNF vector

Abstract

An adenoviral vector comprising (a) an adenoviral genome deficient in the E4 region of the adenoviral genome, (b) a nucleic acid sequence coding for TNF, and (c) a radiation inducible promoter operably linked to the nucleic acid sequence coding for TNF. This invention also provides an adenoviral vector comprising (a) an adenoviral genome deficient in the E4 region of the adenoviral genome, (b) a nucleic acid sequence coding for TNF, and (c) a spacer element of at least 15 base pairs in the E4 region of the adenoviral genome. A method of producing an adenoviral vector and a method of treating a tumor or cancer in a host comprising administering an anti-cancer or anti-tumor effective amount of the adenoviral vector of the present invention also is provided.

What is claimed is:  
     
         1 . An adenoviral vector comprising (a) an adenoviral genome deficient in the E4 region of the adenoviral genome, and optionally deficient in the E1 region, the E2A region, and/or the E3 region of the adenoviral genome, (b) a nucleic acid sequence coding for TNF, and (c) a radiation inducible promoter operably linked to the nucleic acid sequence coding for TNF.  
     
     
         2 . The adenoviral vector of  claim 1 , wherein the adenoviral genome is deficient in the E1 region of the adenoviral genome.  
     
     
         3 . The adenoviral vector of  claim 2 , wherein the nucleic acid sequence coding for TNF is located in the E1 region of the adenoviral genome.  
     
     
         4 . The adenoviral vector of  claim 3 , wherein the TNF is TNF-α.  
     
     
         5 . The adenoviral vector of  claim 3 , wherein the nucleic acid sequence coding for TNF further comprises a transcription-terminating region.  
     
     
         6 . The adenoviral vector of  claim 1 , wherein the radiation inducible promoter is Egr-1.  
     
     
         7 . The adenoviral vector of  claim 2 , wherein the adenoviral vector is deficient in the E3 region of the adenoviral genome.  
     
     
         8 . The adenoviral vector of  claim 7 , wherein the adenoviral genome is deficient in the entire E3 region.  
     
     
         9 . The adenoviral vector of  claim 1 , wherein the adenoviral genome is deficient in the entire coding region of the E4 region.  
     
     
         10 . The adenoviral vector of  claim 9 , wherein the coding region of the E4 region of the adenoviral genome has been replaced with a spacer element having at least 15 base pairs.  
     
     
         11 . The adenoviral vector of  claim 10 , wherein the spacer element is transcriptionally inert.  
     
     
         12 . The adenoviral vector of  claim 11 , wherein the spacer element comprises a polyadenylation sequence that is non-native to the E4 region of the adenoviral genome.  
     
     
         13 . The adenoviral vector of  claim 12 , wherein the spacer element comprises an SV40 polyadenylation sequence.  
     
     
         14 . The adenoviral vector of  claim 10 , wherein the viral production level of the adenoviral vector is greater than the viral production level of the adenoviral vector without the spacer element.  
     
     
         15 . The adenoviral vector of  claim 1 , wherein the adenoviral genome is a serotype-5 adenoviral genome.  
     
     
         16 . A replication competent adenovirus-free stock of the adenoviral vector of  claim 1 .  
     
     
         17 . A host cell comprising the adenoviral vector of  claim 1 .  
     
     
         18 . An adenoviral vector comprising (a) an adenoviral genome deficient in the E4 region of the adenoviral genome, and optionally deficient in the E1 region, the E2A region, and/or the E3 region of the adenoviral genome, (b) a nucleic acid sequence coding for a TNF which is secreted from a cell infected with the adenoviral vector, and (c) a spacer element of at least 15 base pairs in the E4 region of the adenoviral genome.  
     
     
         19 . The adenoviral vector of  claim 18 , wherein the adenoviral genome is deficient in the E1 region of the adenoviral genome.  
     
     
         20 . The adenoviral vector of  claim 18 , wherein the spacer element is transcriptionally inert.  
     
     
         21 . The adenoviral vector of  claim 20 , wherein the spacer element comprises a polyadenylation sequence that is non-native to the E4 region of the adenoviral genome.  
     
     
         22 . The adenoviral vector of  claim 21 , wherein the spacer element comprises an SV40 polyadenylation sequence.  
     
     
         23 . The adenoviral vector of  claim 19 , wherein the nucleic acid sequence coding for TNF is located in the E1 region of the adenoviral genome.  
     
     
         24 . The adenoviral vector of  claim 23 , wherein the TNF is TNF-α.  
     
     
         25 . The adenoviral vector of  claim 23 , wherein the nucleic acid sequence coding for TNF further comprises a transcription-terminating region.  
     
     
         26 . The adenoviral vector of  claim 18 , wherein the adenoviral vector is deficient in the E3 region of the adenoviral genome.  
     
     
         27 . The adenoviral vector of  claim 26 , wherein the adenoviral genome is deficient in the entire E3 region.  
     
     
         28 . The adenoviral vector of  claim 18 , wherein the adenoviral genome is deficient in the entire coding region of the E4 region.  
     
     
         29 . The adenoviral vector of  claim 18 , wherein the viral production level of the adenoviral vector is greater than the viral production level of the adenoviral vector without the spacer element.  
     
     
         30 . The adenoviral vector of  claim 18 , wherein the adenoviral genome is a serotype-5 adenoviral genome.  
     
     
         31 . A replication competent adenoviral-free stock of the adenoviral vector of  claim 18 .  
     
     
         32 . A host cell comprising the adenoviral vector of  claim 18 .  
     
     
         33 . A method of treating a tumor or cancer in a mammal comprising administering an anti-tumor or anti-cancer effective amount of the adenoviral vector of  claim 1  directly to the tumor or cancer of the mammal.  
     
     
         34 . The method of  claim 33 , further comprising the administration of radiation to the host.  
     
     
         35 . The method of  claim 34 , wherein the radiation induces expression of the nucleic acid sequence coding for TNF to produce a therapeutic level of TNF in the host.  
     
     
         36 . The method of  claim 35 , wherein the administration of radiation comprises the use of an internal source of radiation.  
     
     
         37 . The method of  claim 33 , further comprising the administration of a TNF antagonist to the host.  
     
     
         38 . The method of  claim 37 , wherein the TNF antagonist is at least one selected from the group comprising soluble TNF receptors and anti-TNF anti-bodies.  
     
     
         39 . A method of treating a tumor or cancer in a mammal comprising administering an anti-tumor or anti-cancer effective amount of the adenoviral vector of  claim 18  directly to the tumor or cancer of the mammal.  
     
     
         40 . A method of producing an adenoviral vector comprising (a) providing an adenoviral genome that is deficient in the E4 region of the adenoviral genome, (b) inserting a nucleic acid sequence coding for TNF into the adenoviral genome, and (c) inserting a radiation-inducible promoter into the adenoviral genome such that it is operably linked to the nucleic acid sequence coding for TNF.  
     
     
         41 . The method of  claim 40 , wherein the adenoviral genome comprises a spacer element having at least 15 base pairs in the E4 region of the adenoviral genome.  
     
     
         42 . The method of  claim 40 , wherein the radiation inducible promoter is Egr-1.  
     
     
         43 . The adenoviral vector of  claim 1 , wherein the nucleic acid sequence encodes a TNF which is secreted from a cell infected with the adenoviral vector.  
     
     
         44 . The adenoviral vector of  claim 1 , wherein the nucleic acid sequence coding for TNF comprises SEQ ID NO: 2.  
     
     
         45 . The adenoviral vector of  claim 18 , wherein the nucleic acid sequence coding for a TNF which is secreted from a cell infected with the adenoviral vector comprises SEQ ID NO: 2.  
     
     
         46 . The method of  claim 33 , wherein the adenoviral vector is administered to a tumor in a mammal.  
     
     
         47 . The method of  claim 46 , wherein the mammal is a human.  
     
     
         48 . The method of  claim 39 , wherein the adenoviral vector is administered to a tumor in a mammal.  
     
     
         49 . The method of  claim 48 , wherein the mammal is a human.  
     
     
         50 . A pharmaceutical composition comprising the adenoviral vector of  claim 1  and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition does not contain replication-competent adenoviruses.  
     
     
         51 . A pharmaceutical composition comprising the adenoviral vector of  claim 18  and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition does not contain replication-competent adenoviruses.