Prohibitin as Target for Cancer Therapy

Abstract

The present invention relates to pharmaceutical compositions comprising inhibitors of Prohibitin (PHB) for the prevention or/and treatment of hyperproliferative disorders.

1 - 38 . (canceled) 
     
     
         39 . Use of at least one inhibitor of Prohibitin (PHB) optionally together with pharmaceutically acceptable carriers, adjuvants, diluents or/and additives for the manufacture of a pharmaceutical composition for the treatment or/and prevention of hyperprolerative disorders. 
     
     
         40 . Use as claimed in  claim 39 , wherein the pharmaceutical composition is for the treatment or/and prevention of tumours, metastatic tumours, benign tumours, carcinoma, neoplastic carcinoma, gastric carcinoma, larynx carcinoma, neoplastic thyroid cancer, hepatocellular carcinoma, hyperplasia, adenocarcinoma, bladder carcinoma, EGFR overexpressing tumours, Her-2 family overexpressing tumours, Herceptin resistant tumours, B-Raf transformed tumours, or/and Raf-1 transformed tumours. 
     
     
         41 . Use as claimed in  claim 39 , wherein the at least one inhibitor specifically inhibits the interaction of PHB with a Raf kinase, in particular the binding of PHB to a Raf kinase, preferably Raf-1. 
     
     
         42 . Use as claimed in  claim 39 , wherein the at least one inhibitor inhibits activation of the Ras-Raf signalling pathway, preferably the Ras-Raf-MAPK pathway or/and the PHB dependent branch of the Ras signalling pathway. 
     
     
         43 . Use as claimed in  claim 39 , wherein the at least one inhibitor specifically disrupts a Ras-Raf interaction. 
     
     
         44 . Use as claimed in  claim 39 , wherein the at least one inhibitor inhibits downstream of members of the EGFR family, in-2-particular downstream of EGFR and Her-2. 
     
     
         45 . Use as claimed in  claim 39 , wherein the at least one inhibitor converts tumour cells from a transformed to a non-transformed phenotype at least in vitro and in vivo. 
     
     
         46 . Use as claimed in  claim 39 , wherein the at least one inhibitor prevents angiogenesis in tumours. 
     
     
         47 . Use as claimed in  claim 39 , wherein the at least one inhibitor prevents cell migration in vitro or/and in vivo. 
     
     
         48 . Use as claimed in  claim 39 , wherein the at least one inhibitor downregulates PHB transcription or/and translation, inhibits targeting of PHB to membranes, inhibits targeting of a Raf kinase, in particular Raf-1 (C-Raf) to membranes, or/and inhibits posttranslational modification of PHB, in particular required for the PHB targeting to membranes or/and required for the targeting of a Raf kinase, in particular Raf-1 (C-Raf) to membranes. 
     
     
         49 . Use as claimed in  claim 39 , wherein the at least one inhibitor is selected from the group of nucleic acids, nucleic acid analogues such as ribozymes, peptides, polypeptides, and antibodies. 
     
     
         50 . Use as claimed in  claim 49 , wherein the nucleic acid is
 (i) an RNA molecule capable of RNA interference,   (ii) a precursor of the RNA molecule (i), or   (iii) a DNA molecule encoding the RNA molecule (i) or the precursor (ii).   
     
     
         51 . Use as claimed in  claim 50 , wherein the DNA molecule is a vector. 
     
     
         52 . Use as claimed in  claim 49 , wherein the RNA molecule is a double-stranded RNA molecule, preferably a double-stranded siRNA molecule with or without a single-stranded overhang alone at one end or at both ends. 
     
     
         53 . Use as claimed in  claim 49 , wherein the nucleic acid is an antisense nucleic acid. 
     
     
         54 . Use as claimed in  claim 49 , wherein the peptide or/and the polypeptide is biologically inactive, capable of inhibiting the activity of PHB, and comprises a fragment of SEQ.ID.NO:2, a derivative thereof, a fragment to of a Raf kinase or/and a derivative thereof. 
     
     
         55 . Use as claimed in  claim 49 , wherein the antibody is directed against PHB. 
     
     
         56 . Use as claimed in  claim 55 , wherein the antibody is directed against a peptide or polypeptide comprising
 (a) the amino acid sequence of SEQ.ID.NO:2,   (b) an amino acid sequence which is at least 70%, preferably at least 80%, more preferably at least 90% homologous to the sequence of (a), or/and   (c) an immunogenic fragment of (a) or (b).   
     
     
         57 . A nucleic acid suitable as inhibitor in a pharmaceutical composition as defined in  claim 39  comprising
 (a) a fragment of the nucleotide sequence of SEQ.ID.NO:1, or   (b) a fragment which is at least 70%, preferably at least 80%, more preferably at least 90% homologous to the sequence of (a).   
     
     
         58 . The nucleic acid as claimed in  claim 57  having a length of at least 15, preferably at least 17, more preferably at least 19, most preferably at least 21 nucleotides. 
     
     
         59 . The nucleic acid as claimed in  claim 57 , which has a length of at the maximum 29, preferably at the maximum 27, more preferably at the maximum 25, especially more preferably at the maximum 23, most preferably at the maximum 21 nucleotides. 
     
     
         60 . The nucleic acid as claimed in  claim 57 , which is an RNA molecule, preferably a double-stranded RNA molecule, more preferably a double-stranded siRNA molecule with or without a single-stranded overhang alone at one end or at both ends. 
     
     
         61 . The nucleic acid as claimed in  57  comprising a sequence selected from the group consisting of SEQ.ID.NO:5, SEQ.ID.NO:6, SEQ.ID.NO:7, SEQ.ID,NO:8, SEQ.ID.NO;9, SEQ,ID.NO:10 and fragments thereof which may have a length of at least 15 nucleotides. 
     
     
         62 . A screening method for identification of a compound suitable as inhibitor in a pharmaceutical composition defined in  claim 39 , comprising the steps
 (a) providing a cell or/and a transgenic non-human animal capable of expressing, particularly overexpressing PHB,   (b) contacting a compound with the cell or/and the transgenic non-human animal,   (c) determining the amount or/and the activity of PHB, and   (d) selecting a compound which reduces the amount or/and the activity of PHB.   
     
     
         63 . The screening method as claimed in  claim 62 , wherein the activity of PHB in steps (c) and (d) is determined by measuring PHB expression, by prevention of cell migration, by determining targeting of PHB to membranes, by determining targeting of a Raf kinase, in particular Raf-1 to membranes, or/and by determining posttranslational modification of PHB. 
     
     
         64 . The screening method as claimed in  claim 62 , wherein the activity of PHB in steps (c) and (d) is an interaction of PHB with a Raf kinase, in particular the binding of PHB to a Raf kinase, preferably Raf-1, or/and targeting of a Raf kinase, preferably Raf-1, to membranes. 
     
     
         65 . A screening method for identification of a compound suitable as inhibitor in a pharmaceutical composition defined in  claim 62 , comprising the steps
 (a) contacting a compound with isolated PHB or/and an isolated Raf kinase,   (b) determining the activity of PHB to interact with Raf kinase, and   (c) selecting a compound which reduces the activity of PHB.   
     
     
         66 . The screening method as claimed in  claim 65 , wherein the interaction is the binding of PHB to a Raf kinase, preferably Ref-1 or/and targeting of a Raf kinase, preferably Raf-1, to membranes. 
     
     
         67 . A method for treating or/and prevention of hyperprolerative disorders comprising the administration of a pharmaceutical composition of  claim 39  to a subject in need thereof. 
     
     
         68 . Use of an inhibitor of Prohibitin for prevention of cell migration in vivo or/and in vitro. 
     
     
         69 . A screening method for identification of genes suitable as targets for treatment of a hyperproliferative disorder, comprising the steps
 (a) providing a cell or/and a transgenic non-human animal capable of so expressing, particularly overexpressing PHB,   (b) downregulating PHB expression or/and inhibiting PHB activity within the cell or/and the transgenic non-human animal,   (c) determining the expression or/and activity of at least one gene within the cell or/and the animal, and   (d) identifying a target gene of which the expression is upregulated or downregulated by downregulation of PHB expression or/and inhibition of PHB activity.   
     
     
         70 . The screening method of  claim 69  wherein the downregulation of PHB expression or/and the inhibition of PHB activity in step (b) is performed by an inhibitor of PHB. 
     
     
         71 . The screening method of  claim 69 , wherein in step (d) the target gene is upregulated or downregulated by a factor of at least 2, preferably at least 4. 
     
     
         72 . The screening method of  claim 69 , wherein the expression of the at least one gene in step (c) is determined by a microarray technique. 
     
     
         73 . A screening method for identification of a compound suitable for treatment of a hyperproliferative disorder comprising the steps
 (a) providing a cell or/and a transgenic non-human animal capable of expressing at least one gene selected from Table 1 or/and Table 2,   (b) contacting at least one compound with the cell or/and the transgenic non-human animal,   (c) determining the amount or/and the activity of the gene product of the at least one gene selected from Table 1 or/and Table 2, and   (d) identifying a compound which increases the amount or/and the activity of the gene product of the at least one gene of Table 1 or decreases the amount or/and activity of the gene product of the at least one gene of Table 2.   
     
     
         74 . The screening method of  claim 69 , wherein the hyperproliferative disorder is selected from tumours, metastatic tumours, benign tumours, carcinoma, neoplastic carcinoma, gastric carcinoma, larynx carcinoma, neoplastic thyroid cancer, hepatocellular carcinoma, hyperplasia, adenocarcinoma, bladder carcinoma, EGFR overexpressing tumours, Her-2 family overexpressing tumours, Herceptin resistant tumours, B-Raf transformed tumours, and Raf-1 (C-Raf) transformed tumours. 
     
     
         75 . The screening method of  claim 69 , wherein in step (a) the cell is a mammalian cell. 
     
     
         76 . The screening method wherein the downregulation of PHB expression or/and the inhibition of PHB activity in step (b) is performed by an inhibitor as defined in  claim 39 .