Methods for Reversing Multiple Resistance in Animal Cells

Abstract

The present invention is related to the use of a virus, preferably an adenovirus for reversing resistance in cells.

1 - 2 . (canceled) 
     
     
         3 . A method for restoring drug sensitivity of cells in a subject comprising the step of administering an effective amount of an adenovirus to said subject, whereby the adenovirus is replicating in a YB-1 dependent manner. 
     
     
         4 - 5 . (canceled) 
     
     
         6 . The method according to  claim 3 , whereby the cells are tumor cells. 
     
     
         7 - 9 . (canceled) 
     
     
         10 . The method according to  claim 3 , characterised in that the resistance is mediated by an ABC transporter. 
     
     
         11 . The method according to  claim 10 , characterised in that the resistance is a multiple resistance or polyresistance, particular a multiple or polyresistance against cytostatics and/or radiation. 
     
     
         12 . (canceled) 
     
     
         13 . The method according to  claim 3 , whereby the method also comprises the step of administering to said subject a further pharmaceutically active agent, or radiation. 
     
     
         14 . (canceled) 
     
     
         15 . The method according to  claim 10 , whereby the ABC transporter is selected from the group comprising MRP and MDR, in particular MDR-1. 
     
     
         16 - 17 . (canceled) 
     
     
         18 . The method according to  claim 13  characterised in that the adenovirus is administered about 1 to 3 days, preferably about 1 to 2 days prior to the administration of the further pharmaceutically active agent or prior to the radiation. 
     
     
         19 . (canceled) 
     
     
         20 . The method according to  claim 13 , characterised in that the further pharmaceutically active agent is selected from the group comprising cytostatics. 
     
     
         21 - 25 . (canceled) 
     
     
         26 . The method according to  claim 3 , characterised in that the adenovirus is E1A minus. 
     
     
         27 . The method according to  claim 3 , characterized in that the adenovirus is an oncolytic adenovirus. 
     
     
         28 . The method according to  claim 3 , whereby the adenovirus, is replication deficient in cells which lack YB-1 in the nucleus, and whereby the virus encodes an oncogene or oncogene product, in particular an oncogene protein, which transactivates at least one viral gene, preferably an adenoviral gene, whereby the gene is selected from the group comprising Et B55kDa, E4orf6, E4orf3 and E3ADP. 
     
     
         29 . The method according to  claim 28 , characterised in that the adenovirus replicates in cells which have YB-1 in the nucleus. 
     
     
         30 . The method according to  claim 28 , characterised in that the viral oncogene protein is E1A and/or the oncogene is the gene coding for E1A and/or the oncogene protein E1A. 
     
     
         31 - 32 . (canceled) 
     
     
         33 . The method according to  claim 30 , characterized in that the viral oncoprotein E1A does not induce the localization of YB-1 into the nucleus. 
     
     
         34 - 36 . (canceled) 
     
     
         37 . The method according to  claim 28 , characterised in that the oncogene protein exhibits one or several mutations or deletions compared to the wildtype oncogene protein E1A, whereby the deletion is preferably one selected from the group comprising deletions of the CR3 stretches and deletions of the N-terminus and deletions of the C-terminus. 
     
     
         38 - 41 . (canceled) 
     
     
         42 . The method according to  claim 28 , characterized in that the adenovirus codes for YB-1. 
     
     
         43 . The method according to  claim 42 , characterised in that YB-1 is under the control of a tissue-specific and/or tumor-specific promoter. 
     
     
         44 - 45 . (canceled) 
     
     
         46 . The method according to  claim 29 , characterised in that the tumor comprises YB-1 in the nucleus after induction of the transport of YB-1 into the nucleus. 
     
     
         47 . The method according to  claim 46 , characterised in that the transport of YB-1 into the nucleus is triggered by at least one measure selected from the group comprising irradiation, administration of cytostatics and hyperthermia. 
     
     
         48 . (canceled) 
     
     
         49 . The method according to  claim 28 , characterised in that the virus, preferably the adenovirus, is selected from the group comprising AdΔ24, dl922-947, E1Ad/01/07, dl1119/1131, CB 016, dl520 and viruses lacking an expressed viral oncogene which is capable of binding a functional Rb tumor suppressor gene product. 
     
     
         50 . (canceled) 
     
     
         51 . The method according to  claim 28 , whereby the virus comprises a nucleic acid coding for a transgene. 
     
     
         52 - 53 . (canceled) 
     
     
         54 . The method according to  claim 51 , whereby the transgene is selected from the group comprising prodrug genes, cytokines and genes for cytokines, apoptosis-inducing genes, tumor suppressor genes, genes for metalloproteinase inhibitors and genes for angiogenesis inhibitors. 
     
     
         55 . The method according to  claim 51 , whereby the transgene is selected from the group comprising nucleic acids for siRNA, for aptamers, for antisense molecules and for ribozymes, whereby the siRNA, the aptamer, the antisense molecule and/or the ribozyme are targeting a target molecule. 
     
     
         56 - 57 . (canceled) 
     
     
         58 . The method according to  claim 13 , whereby the pharmaceutically active agent is selected from the group comprising cytokines, metalloproteinase inhibitors, angiogenesis inhibitors, cytostatic s, cell cycle inhibitors, proteosome inhibitors, recombinant antibodies, inhibitors of the signal transduction pathway and inhibitors of protein kinases. 
     
     
         59 - 72 . (canceled) 
     
     
         73 . The method according to  claim 3 , whereby the virus, preferably the adenovirus expresses a first protein which is selected from the group comprising an E1B protein and an E4 protein, prior to a second protein which is selected from the group comprising an E1A-protein. 
     
     
         74 . The method according to  claim 73 , characterised in that the first protein is an E1B protein, preferably an E1B55kd protein. 
     
     
         75 . The method according to  claim 73 , characterised in that the first protein is an E4 protein, preferably an E4orf6 protein. 
     
     
         76 . The method according to  claim 73 , characterised in that the first protein is a combination of E1B protein and E4 protein, preferably a combination of E1B55kD protein and E4orf6 protein. 
     
     
         77 . The method according to  claim 73 , characterised in that the E1A protein is an E1A12S protein. 
     
     
         78 - 85 . (canceled) 
     
     
         86 . The method according to  claim 73 , characterised in that the expression of the E1B protein is controlled by a promoter, whereby the promoter is selected from the group comprising tumor-specific promoters, organ-specific promoters, tissue-specific promoters, heterologous promoters and adenoviral promoters, whereby the adenoviral promoter is different from the E1B promoter. 
     
     
         87 . The method according to  claim 73 , characterised in that the expression of the E4 protein is controlled by a promoter, whereby the promoter is selected from the group comprising tumor-specific promoters, organ-specific promoters, tissue-specific promoters, heterologous promoters and adenoviral promoters, whereby the adenoviral promoter is different from the E4 promoter. 
     
     
         88 . (canceled) 
     
     
         89 . The method according to  claim 73 , characterised in that the expression of the E1A protein is controlled by a promoter, whereby the promoter is selected from the group comprising tumor-specific promoters, organ-specific promoters, tissue-specific promoters, heterologous promoters and adenoviral promoters, whereby the adenoviral promoter is different from the E1A promoter. 
     
     
         90 . The method according to  claim 89 , characterised in that the expression of the E1A protein is YB-1 controlled or can be regulated by YB-1. 
     
     
         91 . The method according to  claim 90 , characterised in that the promoter controlling the expression of the E1A protein is the adenoviral E2 late promoter. 
     
     
         92 - 122 . (canceled) 
     
     
         123 . The method according to  claim 73 , characterised in that the virus is replication deficient. 
     
     
         124 . The method according to  claim 123 , characterised in that the virus is capable of replicating in cells comprising deregulated YB-1 or having YB-1 in the nucleus. 
     
     
         125 - 140 . (canceled) 
     
     
         141 . The method according to  claim 3 , characterised in that the adenovirus comprises:
 a lacking functional wildtype E1 region, and   a transporter for the transport of YB-1 into the nucleus of a cell which is infected by the virus.   
     
     
         142 . (canceled) 
     
     
         143 . The method according to  claim 141 , characterised in that the lacking functional wildtype E1A region is E1A-minus. 
     
     
         144 . (canceled) 
     
     
         145 . The method according to  claim 141 , characterised in that the lacking functional wildtype E1 region is E1B55k-minus and/or E1B19k-minus and/or protein IX-minus. 
     
     
         146 - 147 . (canceled) 
     
     
         148 . The method according to  claim 141 , characterised in that the transporter comprises the protein E4orf6. 
     
     
         149 . The method according to  claim 141 , characterised in that the transporter comprises the protein E1B55k. 
     
     
         150 . The method according to  claim 141 , characterised in that the transporter comprises a complex consisting of E4orf4 and E1B55k. 
     
     
         151 - 162 . (canceled) 
     
     
         163 . The method according to  claim 141 , characterised in that the lacking functional wildtype E1 region is E1A13S-minus and/or E1A12S-minus. 
     
     
         164 - 165 . (canceled) 
     
     
         166 . The method according to  claim 141 , characterised in that the adenovirus comprises a heterologous nucleic acid coding for the E1A12S protein and that said nucleic acid is under the control of a promoter, whereby the promoter is preferably a YB-1-dependent promoter and is more preferably selected from the group comprising the E2-late promoter, tumor-specific promoters and tissue-specific promoters. 
     
     
         167 - 170 . (canceled) 
     
     
         171 . The method according to  claim 141 , characterised in that the virus adenovirus comprises a nucleic acid coding for YB-1. 
     
     
         172 - 201 . (canceled) 
     
     
         202 . The method according to  claim 13 , whereby the administration of the adenovirus causes or increases the efficacy of the further pharmaceutically active agent and/or of the radiation.