Circular dumbbell decoy oligodeoxynucleotides (cdodn) containing dna bindings sites of transcription

Abstract

The present invention provides a circular dumbbell oligodeoxynucleotide (CDODN) comprising two loop structures and a stem structure, wherein the stem structure comprises a nucleotide sequence capable of binding the DNA-binding domain of a transcriptional factor. The present invention further provides a pharmaceutical composition comprising said CDODN. The pharmaceutical composition can be used for treating and/or preventing a disease or disorder related to such a transcriptional factor. The present invention also provides a method for treating and/or preventing a disease or disorder related to such a transcriptional factor, comprising administering to the subject a therapeutically effective amount of a CDODN comprising two loop structures and a stem structure, wherein the stem structure comprises a nucleotide sequence capable of binding the DNA-binding domain of the transcriptional factor.

1 . A circular dumbbell oligodeoxynucleotide (CDODN) comprising two loop structures and a stem structure, wherein the stem structure comprises a nucleotide sequence capable of binding the DNA-binding domain of a transcriptional factor.  
     
     
         2 . The CDODN according to  claim 1 , wherein said transcriptional factor is selected from the group consisting of NFκB, STAT-1, GATA-3, STAT-6, AP-1, E2F, Ets and CRE.  
     
     
         3 . The CDODN of  claim 1 , which comprises two identical stem loop structures covalently linked by enzymatic ligation.  
     
     
         4 . The CDODN of  claim 1 , which does not contain any chemically modified nucleotides.  
     
     
         5 . The CDODN of  claim 1 , which stem structure additionally comprises nucleotide sequences capable of binding the DNA-binding domain of two or more transcription factors.  
     
     
         6 . The CDODN according to  claim 1 , said transcriptional factor is AP-1.  
     
     
         7 . The CDODN of  claim 6 , wherein the nucleotide sequence capable of binding the DNA-binding domain of AP-1 is 5′-TGACTCA-3′.  
     
     
         8 . The CDODN of  claim 6 , wherein each of the identical stem loop structures has the sequence of SEQ ID. NO. 3.  
     
     
         9 . The CDODN of  claim 6 , which stem structure additionally comprises a nucleotide sequence capable of binding the DNA-binding domain of another transcription factor.  
     
     
         10 . The CDODN of  claim 6 , which has an AP-1 sequence specificity of greater than about 5 times that of a phosphorothiolated oligonucleotide with the sequence of SEQ ID. NO. 4, as assessed by in vitro competitive binding assay.  
     
     
         11 . The CDODN of  claim 1 , wherein said transcriptional factor is E2F.  
     
     
         12 . The CDODN of  claim 11 , wherein the nucleotide sequence capable of binding the DNA-binding domain of E2F is 5′-TTTCGCGC-3′.  
     
     
         13 . The CDODN of  claim 11 , wherein each of the identical stem loop structures has the sequence of SEQ ID. NO. 6.  
     
     
         14 . The CDODN of  claim 11 , which has an E2F sequence specificity of greater than about 5 times that of a phosphorothiolated oligonucleotide with the sequence of SEQ ID. NO. 7, as assessed by in vitro competitive binding assay.  
     
     
         15 . The CDODN of  claim 1 , wherein said transcriptional factor is NFκB.  
     
     
         16 . A method for treating or preventing a disease or disorder related to a transcriptional factor in a subject, comprising administering to the subject a therapeutically effective amount of a CDODN comprising two loop structures and a stem structure, wherein the stem structure comprises a nucleotide sequence capable of binding the DNA-binding domain of the transcriptional factor.  
     
     
         17 . The method according to  claim 16 , wherein said transcriptional factor is selected from the group consisting of NFκB, STAT-1, GATA-3, STAT-6, AP-1, E2F, Ets and CRE.  
     
     
         18 . The method of  claim 16 , where the pharmaceutically acceptable carrier is a HVJ-liposome composition.  
     
     
         19 . The method according to  claim 16 , wherein said transcriptional factor is AP-1.  
     
     
         20 . The method according to  claim 18 , wherein the disease or disorder related to a transcriptional factor comprises vascular smooth muscle cell proliferation or neointimal hyperplasia in the subject following vessel injury.  
     
     
         21 . The method of  claim 19 , where the amount of the CDODN is sufficient to prevent restenosis in the subject.  
     
     
         22 . The method of  claim 20 , where the compound is administered prior to the vessel injury.  
     
     
         23 . The method of  claim 16 , wherein said transcriptional factor is E2F.  
     
     
         24 . The method of  claim 23 , wherein said disease or disorder comprises vascular smooth muscle cell proliferation or neointimal hyperplasia in the subject following vessel injury.  
     
     
         25 . The method of  claim 23 , wherein the therapeutically effective amount of the CDODN is effective to prevent restenosis in the subject.  
     
     
         26 . The method of  claim 24 , wherein the CDODN is administered after the vessel injury.  
     
     
         27 . The method of  claim 16 , wherein said transcriptional factor is NFκB.  
     
     
         28 . The method of  claim 16 , wherein said disease or disorder comprises inflammatory bowel disease.  
     
     
         29 . A pharmaceutical composition for treating or preventing a disease or disorder related to a transcriptional factor in a subject, comprising a therapeutically effective amount of a CDODN comprising two loop structures and a stem structure, wherein the stem structure comprises a nucleotide sequence capable of binding the DNA-binding domain of the transcriptional factor, and a pharmaceutically acceptable carrier.  
     
     
         30 . The pharmaceutical composition according to  claim 29 , wherein said transcriptional factor is selected from the group consisting of NFκB, STAT-1, GATA-3, STAT-6, AP-1, E2F, Ets and CRE.  
     
     
         31 . The pharmaceutical composition of  claim 29 , where the pharmaceutically acceptable carrier is a HVJ-liposome composition.  
     
     
         32 . The pharmaceutical composition according to  claim 29 , wherein said transcriptional factor is AP-1.  
     
     
         33 . The pharmaceutical composition according to  claim 29 , wherein said disease or disorder is vascular smooth muscle cell proliferation or neointimal hyperplasia in the subject following vessel injury.  
     
     
         34 . The pharmaceutical composition of  claim 29 , where the amount of the CDODN is sufficient to prevent restenosis in the subject.  
     
     
         35 . The pharmaceutical composition according to  claim 29 , wherein said transcriptional factor is E2F.  
     
     
         36 . The pharmaceutical composition according to  claim 35 , wherein said disease or disorder is vascular smooth muscle cell proliferation or neointimal hyperplasia in the subject following vessel injury.  
     
     
         37 . The pharmaceutical composition of  claim 36 , wherein the therapeutically effective amount of the CDODN is effective to prevent restenosis in the subject.  
     
     
         38 . The pharmaceutical composition of  claim 29 , wherein said transcriptional factor is NFκB.  
     
     
         39 . The pharmaceutical composition  claim 29 , wherein said disease or disorder comprises inflammatory bowel disease.  
     
     
         40 . Use, in the manufacture of a medicament for treating or preventing a disease or disorder related to transcriptional factor in a subject, of a therapeutically effective amount of a CDODN comprising two loop structures and a stem structure, wherein the stem structure comprises a nucleotide sequence capable of binding the DNA-binding domain of the transcriptional factor.  
     
     
         41 . The use according to  claim 40 , wherein said transcriptional factor is selected from the group consisting of NFκB, STAT-1, GATA-3, STAT-6, AP-1, E2F, Ets and CRE.  
     
     
         42 . The use according to  claim 40 , wherein said medicament is in the form of a HVJ-liposome composition.  
     
     
         43 . The use according to  claim 40 , wherein said transcriptional factor is AP-1.  
     
     
         44 . The use according to  claim 43 , wherein said disease or disorder is vascular smooth muscle cell proliferation or neointimal hyperplasia in the subject following vessel injury.  
     
     
         45 . The use according to  claim 43 , wherein the amount of the CDODN is sufficient to prevent restenosis in the subject.  
     
     
         46 . The use according to  claim 40 , wherein said transcriptional factor is E2F.  
     
     
         47 . The use according to  claim 46 , wherein said disease or disorder is vascular smooth muscle cell proliferation or neointimal hyperplasia in the subject following vessel injury.  
     
     
         48 . The use of  claim 40 , wherein the therapeutically effective amount of the CDODN is effective to prevent restenosis in the subject.  
     
     
         49 . The use of  claim 40 , wherein said transcriptional factor is NFκB.  
     
     
         50 . The use  claim 46 , wherein said disease or disorder comprises inflammatory bowel disease.