Antisense-oligonucleotides for the treatment of immuno-suppressive effects of transforming growth factor-beta (TGF-beta)

Abstract

Antisense-oligonucleotides or effective derivatives thereof hybridizing with an area of a gene coding for transforming growth factor-β (TGF-β) comprising the following nucleic acid sequences identified in the sequence listing under SEQ ID NO. 1-56 and 137 or comprising the following nucleic acid sequences identified in the sequence listing under SEQ ID NO. 57 to 136 each of the nucleic acids having a DNA- or RNA-type structure.

1 . Antisense-oligonucleotides or effective derivatives thereof hybridizing with an area of a gene coding for transforming growth factor-β (TGF-β) comprising the following nucleic acid sequences identified in the sequence listing under SEQ ID NO. 1-56 and 137 or comprising the following nucleic acid sequences identified in the sequence listing under SEQ ID NO. 57 to 136 each of the nucleic acids having a DNA- or RNA-type structure.  
     
     
         2 . Antisense-oligonucleotides according to  claim 1  wherein said nucleic acid hybridizes with an area of a gene coding for transforming growth factor-β 1 , -β 2  and/or -β 3 .  
     
     
         3 . Antisense-oligonucleotides according to  claim 1  and/or  2  wherein said nucleic acid is hybridizing with areas of a gene region coding for TGF-β and/or areas of a gene region coding and non-coding for TGF-β.  
     
     
         4 . Antisense-oligonucleotide according to any one of the claims  1 - 3  wherein the antisense-oligonucleotide is a phosphorothioate oligodeoxynucleotide.  
     
     
         5 . Antisense-oligonucleotide according to any one of the  claims 1  to  4  obtainable by solid phase synthesis using phosphite triester chemistry by growing the nucleotide chain in 3′-5′ direction in that the respective nucleotide is coupled to the first nucleotide which is covalently attached to the solid phase comprising the steps of 
 cleaving 5′DMT protecting group of the previous nucleotide,  
 adding the respective nucleotide for chain propagation,  
 modifying phosphite groups subsequently cap unreacted 5′-hydroxyl groups and  
 cleaving the oligonucleotide from the solid support,  
 followed by working up the synthesis product.  
 
     
     
         6 . Antisense-oligonucleotide according to any on of the  claims 1  to  5  wherein the oligonucleotides of the SEQ ID NO. 1 to 56 and 137 are antisense-oligodeoxy-nucleotides of TGF-β 1  and oligonucleotides of the SEQ ID NO. 57 to 136 are phosphorothioate-antisense-oligo-deoxynucleotide of TGF-β 2 .  
     
     
         7 . Oligonucleotide as nonsense-control-nucleotide with an identical amount of GC as the antisense-oligonucleotide according to any one of the  claims 1  to  6 .  
     
     
         8 . Pharmaceutical composition comprising an antisense-nucleic acid according to any one of the  claims 1  to  6 .  
     
     
         9 . Use of antisense-oligonucleotides according to any one of the  claims 1  to  6  for the manufacturing of a pharmaceutical composition of  claim 8  for the treatment of tumors in which expression of TGF-β is of relevance for pathogenicity and/or inhibition of pathological angiogenesis.  
     
     
         10 . Use according to  claim 9  for manufacturing pharmaceutical composition for the treatment of the immunosuppressive effect of TGF-β, augmentation of the proliferation of cytotoxic lymphocytes, for the treatment of endogenous hyper expression of TGF-β, for treatment of breast tumors, for treatment of neuro-fibroma, malignant glioma inculding glioblastoma for the treatment and prophylaxis of skin carcinogenesis as well as treatment of esophageal and gastric carcinomas.  
     
     
         11 . Method of treating tumors in which the expression of TGF-β is of relevance for pathogenicity by inhibiting TGF-β and thereby reducing an immuno suppression and/or inhibition of pathological angiogenesis by administration of an effective dose of an antisense-TGF-β-oligonucleotides according to any one of the  claims 1  to  6 .  
     
     
         12 . Method according to  claim 11  wherein the reduction of immuno suppression is accompanied by an augmented proliferation of cytotoxic lymphocytes in comparison with the status before administration of the antisense-TGF-β-oligonucleotides and thereupon starting cytotoxic activity decreases the numbers of tumor cells.