Peptides that deliver antisense oligonucleotides which downregulate protein expression in cells

Abstract

The present invention provides a peptide that can complex with an oligonucleotide, e.g. an antisense oligonucleotide, and deliver it into a cell. The present invention also provides compositions comprising a complex of such a peptide and an oligonucleotide and methods of delivering an oligonucleotide into a cell and of inhibiting protein expression using such compositions. The present invention also provides a method of making such a complex, a method of sensitizing cells to anti-cancer agents such as paclitaxel, and pharmaceutical compositions.

1 .- 10 . (canceled)  
     
     
         11 . A peptide comprising consecutive amino acids, the sequence of which amino acids is shown in SEQ ID NO: 1.  
     
     
         12 . (canceled)  
     
     
         13 . A composition comprising a complex between the peptide of  claim 11  and an oligonucleotide.  
     
     
         14 . The composition of  claim 13 , further comprising an aqueous carrier.  
     
     
         15 . The composition of  claim 13 , wherein the oligonucleotide comprises from about 10 to about 40 consecutive nucleotides.  
     
     
         16 . The composition of  claim 15 , wherein the consecutive nucleotides of the oligonucleotide have a sequence capable of inhibiting translation of a mRNA into a protein.  
     
     
         17 . (canceled)  
     
     
         18 . A method of delivering an oligonucleotide into a cell comprising contacting the cell with the composition of  claim 13 , under conditions permitting the composition to enter the cell and thereby deliver the oligonucleotide into the cell.  
     
     
         19 . A method of inhibiting expression of a protein in a cell comprising delivering an oligonucleotide into the cell using the method of  claim 18 , under conditions permitting the oligonucleotide, once inside the cell, to hybridize with a nucleic acid encoding the protein and thereby inhibit expression of the protein from the nucleic acid in the cell.  
     
     
         20 . The method of  claim 18 , wherein the cell is contacted with a lysosomotropic agent prior to contacting the cell with the composition.  
     
     
         21 . (canceled)  
     
     
         22 . (canceled)  
     
     
         23 . The method of  claim 16 , wherein the sequence of the oligonucleotide is shown in SEQ ID NO:6.  
     
     
         24 . The method of  claim 19 , wherein the protein is Protein Kinase C alpha.  
     
     
         25 . The method of  claim 19 , wherein the cell is of mammalian origin.  
     
     
         26 . (canceled)  
     
     
         27 . (canceled)  
     
     
         28 . The method of  claim 19 , wherein the nucleic acid is a deoxyribonucleic acid.  
     
     
         29 . The method of  claim 19 , wherein the nucleic acid is a ribonucleic acid.  
     
     
         30 . (canceled)  
     
     
         31 . A pharmaceutical composition comprising a therapeutically effective amount of the composition of  claim 13  and a pharmaceutically acceptable carrier.  
     
     
         32 . (canceled)  
     
     
         33 . A method of making a composition, comprising contacting an oligonucleotide with the peptide of  claim 11  under conditions permitting the peptide to form a complex with the oligonucleotide.  
     
     
         34 . A method of increasing the sensitivity of a cancer cell to an anti-cancer agent which comprises inhibiting expression of a protein in the cancer cell using the method of  claim 19 .  
     
     
         35 . The method of  claim 34 , wherein the anti-cancer agent is paclitaxel.  
     
     
         36 . The method of  claim 35 , wherein the protein is protein kinase C alpha.  
     
     
         37 . The method of  claim 36 , wherein the cancer cell is a bladder cancer cell.  
     
     
         38 . (canceled)  
     
     
         39 . A method of delivering an oligonucleotide into a cell comprising contacting the cell with the composition of  claim 38 , under conditions permitting the composition to enter the cell and thereby deliver the oligonucleotide into the cell.