Antisense modulation of nuclear hormone receptors

Abstract

Provided are antisense oligonucleotides and other agents that target and modulate nuclear hormone receptors (NHRs) such as the glucocorticoid receptor (GR), compositions that comprise the same, and methods of use thereof.

1 .- 50 . (canceled) 
     
     
         51 . An antisense morpholino oligomer of 20 to 30 bases comprising a base sequence that is complementary to at least 15 contiguous bases of SEQ ID NO:1, wherein the antisense morpholino oligomer alters splicing of the human glucocorticoid receptor (GR) pre-mRNA transcript and increases expression of a ligand independent form of the GR; or a pharmaceutically acceptable salt thereof. 
     
     
         52 . The antisense morpholino oligomer of  claim 51 , wherein the antisense morpholino oligomer has 20 to 23 bases. 
     
     
         53 . The antisense morpholino oligomer of  claim 51 , wherein the base sequence is complementary to at least 17 contiguous bases of SEQ ID NO:1. 
     
     
         54 . The antisense morpholino oligomer of  claim 51 , wherein the base sequence is complementary to at least 20 contiguous bases of SEQ ID NO:1. 
     
     
         55 . The antisense morpholino oligomer of  claim 51 , wherein the base sequence is 100% complementary to contiguous bases of SEQ ID NO:1. 
     
     
         56 . The antisense morpholino oligomer of  claim 51 , wherein the antisense morpholino oligomer contains about 10%-50% intersubunit cationic linkages. 
     
     
         57 . The antisense morpholino oligomer of  claim 51 , wherein the antisense morpholino oligomer comprises an arginine-rich carrier peptide. 
     
     
         58 . The antisense morpholino oligomer of  claim 57 , wherein the peptide is linked at its C-terminus to the 5′ end of the antisense morpholino oligomer through a one or two-amino acid linker. 
     
     
         59 . The antisense morpholino oligomer of  claim 57 , wherein the peptide is linked at its C-terminus to the 3′ end of the antisense morpholino oligomer through a one or two-amino acid linker. 
     
     
         60 . The antisense morpholino oligomer of  claim 59 , wherein the linker is AhxβAla, wherein Ahx is 6-amino hexanoic acid and βAla is β-alanine. 
     
     
         61 . The antisense morpholino oligomer of  claim 51 , wherein the arginine-rich carrier peptide is selected from any one of SEQ ID NOS: 18-26. 
     
     
         62 . The antisense morpholino oligomer of  claim 51 , wherein the antisense morpholino oligomer comprises phosphorus-containing intersubunit linkages in accordance with the structure: 
       
         
           
           
               
               
           
         
         wherein Z═O, Y 1 ═O, and X═N(CH 3 ) 2 . 
       
     
     
         63 . The antisense morpholino oligomer of  claim 56 , wherein the antisense morpholino oligomer comprises phosphorus-containing intersubunit linkages in accordance with the structure: 
       
         
           
           
               
               
           
         
         wherein Z is S or O, 
         X═NR 1 R 2  or OR 6 , 
         Y 1 ═O or NR 7 , 
         and each phosphorus-containing linkage is independently selected from: 
         (a) uncharged linkage (a), wherein each of R 1 , R 2 , R 6 , and R 7  is independently selected from hydrogen and lower alkyl; 
         (b1) cationic linkage (b1), wherein X═NR 1 R 2  and Y 1 ═O, wherein NR 1 R 2  represents a group of the formula: 
       
       
         
           
           
               
               
           
         
         wherein each R is independently H or CH 3 , 
         R 4  is H, CH 3  or an electron pair, and 
         R 3  is selected from H, lower alkyl, C(═NH)NH 2 , Z-L-NHC(═NH)NH 2 , and 
         [C(O)CHR′NH] m H, wherein Z is carbonyl (C(O)) or a direct bond, L is an optional linker up to 18 atoms in length having bonds selected from alkyl, alkoxy, and alkylamino, R′ is a side chain of a naturally occurring amino acid or a one- or two-carbon homolog thereof, and m is 1 to 6; 
         (b2) cationic linkage (b2), wherein X═NR 1 R 2  and Y 1 ═O, wherein R 1 ═H or CH 3 , and R 2 =LNR 3 R 4 R 5 , wherein L, R 3 , and R 4  are defined as above, and R 5  is H, lower alkyl, or lower (alkoxy)alkyl; and 
         (b3) cationic linkage (b3), wherein Y═NR′ and X═OR 6 , wherein R 7 =LNR 3 R 4 R 5 , wherein L, R 3 , R 4 , and R 5  are defined as above, and R 6  is H or lower alkyl; 
         wherein at least one phosphorus-containing intersubunit linkage is selected from cationic linkages (b1), (b2), and (b3). 
       
     
     
         64 . The antisense morpholino oligomer of  claim 63 , wherein each of R 1  and R 2 , in linkages of type (a), is methyl. 
     
     
         65 . The antisense morpholino oligomer of  claim 63 , wherein at least one phosphorus-containing intersubunit linkage is of type (b1), where each R is H, R 4  is H, CH 3 , or an electron pair, and R 3  is selected from H, CH 3 , C(═NH)NH 2 , and C(O)-L-NHC(═NH)NH 2 . 
     
     
         66 . The antisense morpholino oligomer of  claim 63 , wherein at least one phosphorus-containing intersubunit linkage is of type (b1), where each R is H, R 4  is an electron pair, and R 3  is selected from C(═NH)NH 2  and C(O)-L-NHC(═NH)NH 2 . 
     
     
         67 . The antisense morpholino oligomer of  claim 66 , wherein R 3  is C(O)-L-NHC(NH)NH 2 , and L is a hydrocarbon having the structure —(CH 2 ) n —, where n is 1 to 12. 
     
     
         68 . The antisense morpholino oligomer of  claim 63 , wherein at least one phosphorus-containing intersubunit linkage is of type (b1), where each R is H, and each of R 3  and R 4  is independently H or CH 3 . 
     
     
         69 . The antisense morpholino oligomer of  claim 51 , wherein the base sequence comprises any one of SEQ ID NOS:2-14. 
     
     
         70 . The antisense morpholino oligomer of  claim 51 , wherein the base sequence consists of any one of SEQ ID NOS:2-14. 
     
     
         71 . A pharmaceutical composition comprising: (i) an antisense morpholino oligomer of  claim 51  or a pharmaceutically acceptable salt thereof, and (ii) a pharmaceutically acceptable carrier. 
     
     
         72 . An antisense morpholino oligomer of 20 to 30 bases comprising a base sequence that is 100% complementary to contiguous bases of SEQ ID NO:1, wherein the antisense morpholino oligomer comprises phosphorodiamidate intersubunit linkages, and wherein the antisense morpholino oligomer alters splicing of the human glucocorticoid receptor (GR) pre-mRNA transcript and increases expression of a ligand independent form of the GR; or a pharmaceutically acceptable salt thereof. 
     
     
         73 . A pharmaceutical composition comprising: (i) an antisense morpholino oligomer of 20 to 30 bases comprising a base sequence that is 100% complementary to contiguous bases of SEQ ID NO:1, wherein the antisense morpholino oligomer comprises phosphorodiamidate intersubunit linkages, and wherein the antisense morpholino oligomer alters splicing of the human glucocorticoid receptor (GR) pre-mRNA transcript and increases expression of a ligand independent form of the GR; or a pharmaceutically acceptable salt thereof, and (ii) a pharmaceutically acceptable carrier.