Antisense modulation of connective tissue growth factor expression

Abstract

Antisense compounds, compositions and methods are provided for modulating the expression of connective tissue growth factor. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding connective tissue growth factor. Methods of using these compounds for modulation of connective tissue growth factor expression and for treatment of diseases associated with expression of connective tissue growth factor are provided.

1 . A method of preventing or delaying the onset of diabetic nephropathy in an animal, said method comprising administering to said animal an effective amount of a compound 8 to 50 nucleobases in length targeted to a nucleic acid molecule encoding connective tissue growth factor, wherein said compound specifically hybridizes with said nucleic acid molecule encoding connective tissue growth factor and inhibits the expression of connective tissue growth factor.  
     
     
         2 . The method of  claim 1 , wherein said nucleic acid molecule encodes human connective tissue growth factor and has a nucleotide sequence comprising SEQ ID NO: 3, 17, 18 or 19:  
     
     
         3 . The method of  claim 1 , wherein said nucleic acid molecule encodes mouse connective growth factor and has a nucleotide sequence comprising SEQ ID NO: 10 or 98.  
     
     
         4 . The method according to  claim 1 , wherein said compound is an antisense oligonucleotide.  
     
     
         5 . The method according to  claim 4 , wherein said antisense oligonucleotide has a sequence comprising SEQ ID NO: 24, 25, 27, 28, 33, 34, 35, 36, 38, 39, 41, 45, 46, 47, 48, 50, 52, 58, 62, 63, 64, 68, 70, 72, 73, 81, 86, 88, 90, 91, 92, 95, 97, 30, 31, 32, 37, 40, 42, 103, 104, 106, 108, 110, 111, 116, 117, 120, 122, 124, 126, 127, 128, 130, 134, 135, 136, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 151 or 153.  
     
     
         6 . The method according to  claim 4 , wherein said antisense oligonucleotide is a sequence of up to 30 nucleobases in length comprising at least an 8 nucleobase portion of SEQ ID NO: 24, 25, 27, 28, 33, 34, 35, 36, 38, 39, 41, 45, 46, 47, 48, 50, 52, 58, 62, 63, 64, 68, 70, 72, 73, 81, 86, 88, 90, 91, 92, 95, 97, 30, 31, 32, 37, 40, 42, 103, 104, 106, 108, 110, 111, 116, 117, 120, 122, 124, 126, 127, 128, 130, 134, 135, 136, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 151 or 153.  
     
     
         7 . The method according to  claim 4 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 48.  
     
     
         8 . The method according to  claim 4 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 45.  
     
     
         9 . The method according to  claim 4 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 28.  
     
     
         10 . The method according to  claim 4 , wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.  
     
     
         11 . The method according to  claim 10 , wherein the modified internucleoside linkage is a phosphorothioate linkage.  
     
     
         12 . The method according to  claim 4 , wherein the antisense oligonucleotide comprises at least one modified sugar moiety.  
     
     
         13 . The method according to  claim 12 , wherein the modified sugar moiety is a 2′-O-methoxyethyl sugar moiety.  
     
     
         14 . The method according to  claim 4 , wherein the antisense oligonucleotide comprises at least one modified nucleobase.  
     
     
         15 . The method according to  claim 14 , wherein the modified nucleobase is a 5-methylcytosine.  
     
     
         16 . The method according to  claim 4 , wherein the antisense oligonucleotide is a chimeric oligonucleotide.  
     
     
         17 . The method according to  claim 1 , wherein said compound is administered as a composition comprising said compound and a pharmaceutically acceptable carrier or diluent.  
     
     
         18 . The method according to  claim 17 , wherein said composition further comprises a colloidal dispersion system.  
     
     
         19 . The method according to  claim 1 , wherein the animal is a diabetic animal.  
     
     
         20 . The method according to  claim 19 , wherein the diabetic animal has type 1 diabetes.  
     
     
         21 . The method according to  claim 19 , wherein the diabetic animal has type 2 diabetes.  
     
     
         22 . The method according to  claim 1 , wherein the animal is a human or a rodent.  
     
     
         23 . A method of treating or delaying the onset of type 1 or type 2 diabetes in an animal, said method comprising administering to said animal an effective amount of a compound 8 to 50 nucleobases in length targeted to a nucleic acid molecule encoding connective tissue growth factor, wherein said compound specifically hybridizes with said nucleic acid molecule encoding connective tissue growth factor and inhibits the expression of connective tissue growth factor.  
     
     
         24 . The method according to  claim 23 , wherein said compound is an antisense oligonucleotide.  
     
     
         25 . The method according to  claim 24 , wherein said antisense oligonucleotide has a sequence comprising SEQ ID NO: 24, 25, 27, 28, 33, 34, 35, 36, 38, 39, 41, 45, 46, 47, 48, 50, 52, 58, 62, 63, 64, 68, 70, 72, 73, 81, 86, 88, 90, 91, 92, 95, 97, 30, 31, 32, 37, 40, 42, 103, 104, 106, 108, 110, 111, 116, 117, 120, 122, 124, 126, 127, 128, 130, 134, 135, 136, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 151 or 153.  
     
     
         26 . The method according to  claim 24 , wherein said antisense oligonucleotide is a sequence of up to 30 nucleobases in length comprising at least an 8 nucleobase portion of SEQ ID NO: 24, 25, 27, 28, 33, 34, 35, 36, 38, 39, 41, 45, 46, 47, 48, 50, 52, 58, 62, 63, 64, 68, 70, 72, 73, 81, 86, 88, 90, 91, 92, 95, 97, 30, 31, 32, 37, 40, 42, 103, 104, 106, 108, 110, 111, 116, 117, 120, 122, 124, 126, 127, 128, 130, 134, 135, 136, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 151 or 153.  
     
     
         27 . The method according to  claim 24 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 48.  
     
     
         28 . The method according to  claim 24 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 45.  
     
     
         29 . The method according to  claim 24 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 28.  
     
     
         30 . The method according to  claim 24 , wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.  
     
     
         31 . The method according to  claim 30 , wherein the modified internucleoside linkage is a phosphorothioate linkage.  
     
     
         32 . The method according to  claim 24 , wherein the antisense oligonucleotide comprises at least one modified sugar moiety.  
     
     
         33 . The method according to  claim 32 , wherein the modified sugar moiety is a 2′-O-methoxyethyl sugar moiety.  
     
     
         34 . The method according to  claim 24 , wherein the antisense oligonucleotide comprises at least one modified nucleobase.  
     
     
         35 . The method according to  claim 34 , wherein the modified nucleobase is a 5-methylcytosine.  
     
     
         36 . The method according to  claim 24 , wherein the antisense oligonucleotide is a chimeric oligonucleotide.  
     
     
         37 . The method according to  claim 23 , wherein said compound is administered as a composition comprising said compound and a pharmaceutically acceptable carrier or diluent.  
     
     
         38 . The method according to  claim 37 , wherein said composition further comprises a colloidal dispersion system.  
     
     
         39 . The method according to  claim 23 , wherein the animal is a human or a rodent.  
     
     
         40 . A method of inhibiting the expression of connective tissue growth factor in cells or tissues comprising contacting said cells or tissues with a compound 8 to 50 nucleobases in length targeted to a nucleic acid molecule encoding connective tissue growth factor, wherein said compound specifically hybridizes with said nucleic acid molecule encoding connective tissue growth factor and inhibits the expression of connective tissue growth factor.  
     
     
         41 . A method of treating an animal having a disease or condition associated with connective tissue growth factor comprising administering to said animal a therapeutically or prophylactically effective amount of a compound 8 to 50 nucleobases in length targeted to a nucleic acid molecule encoding connective tissue growth factor, wherein said compound specifically hybridizes with said nucleic acid molecule encoding connective tissue growth factor and inhibits the expression of connective tissue growth factor.  
     
     
         42 . The method of  claim 41 , wherein the disease or condition is a hyperproliferative disorder.  
     
     
         43 . The method of  claim 42 , wherein the hyperproliferative disorder is cancer.  
     
     
         44 . The method of  claim 43 , wherein the cancer is selected from the group consisting of breast, prostate and renal cancer.  
     
     
         45 . The method of  claim 41  wherein the disease or condition is selected from the group consisting of pulmonary fibrosis, renal fibrosis, scleroderma, and atherosclerosis.  
     
     
         46 . A method of inhibiting or preventing an increase in proteinuria or albuminuria or both in a diabetic animal, said method comprising administering to said diabetic animal an effective amount of a compound 8 to 50 nucleobases in length targeted to a nucleic acid molecule encoding connective tissue growth factor, wherein said compound specifically hybridizes with said nucleic acid molecule encoding connective tissue growth factor and inhibits the expression of connective tissue growth factor.  
     
     
         47 . The method according to  claim 46 , wherein said compound is an antisense oligonucleotide.  
     
     
         48 . The method according to  claim 47 , wherein said antisense oligonucleotide has a sequence comprising SEQ ID NO: 24, 25, 27, 28, 33, 34, 35, 36, 38, 39, 41, 45, 46, 47, 48, 50, 52, 58, 62, 63, 64, 68, 70, 72, 73, 81, 86, 88, 90, 91, 92, 95, 97, 30, 31, 32, 37, 40, 42, 103, 104, 106, 108, 110, 111, 116, 117, 120, 122, 124, 126, 127, 128, 130, 134, 135, 136, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 151 or 153.  
     
     
         49 . The method according to  claim 47 , wherein said antisense oligonucleotide is a sequence of up to 30 nucleobases in length comprising at least an 8 nucleobase portion of SEQ ID NO: 24, 25, 27, 28, 33, 34, 35, 36, 38, 39, 41, 45, 46, 47, 48, 50, 52, 58, 62, 63, 64, 68, 70, 72, 73, 81, 86, 88, 90, 91, 92, 95, 97, 30, 31, 32, 37, 40, 42, 103, 104, 106, 108, 110, 111, 116, 117, 120, 122, 124, 126, 127, 128, 130, 134, 135, 136, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 151 or 153.  
     
     
         50 . The method according to  claim 47 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 48.  
     
     
         51 . The method according to  claim 47 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 45.  
     
     
         52 . The method according to  claim 47 , wherein said antisense oligonucleotide has a sequence consisting of SEQ ID NO: 28.  
     
     
         53 . The method according to  claim 47 , wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.  
     
     
         54 . The method according to  claim 53 , wherein the modified internucleoside linkage is a phosphorothioate linkage.  
     
     
         55 . The method according to  claim 47 , wherein the antisense oligonucleotide comprises at least one modified sugar moiety.  
     
     
         56 . The method according to  claim 55 , wherein the modified sugar moiety is a 2′-O-methoxyethyl sugar moiety.  
     
     
         57 . The method according to  claim 47 , wherein the antisense oligonucleotide comprises at least one modified nucleobase.  
     
     
         58 . The method according to  claim 57 , wherein the modified nucleobase is a 5-methylcytosine.  
     
     
         59 . The method according to  claim 47 , wherein the antisense oligonucleotide is a chimeric oligonucleotide.  
     
     
         60 . The method according to  claim 46 , wherein said compound is administered as a composition comprising said compound and a pharmaceutically acceptable carrier or diluent.  
     
     
         61 . The method according to  claim 60 , wherein said composition further comprises a colloidal dispersion system.  
     
     
         62 . The method according to  claim 46 , wherein the diabetic animal has type 1 diabetes.  
     
     
         63 . The method according to  claim 46 , wherein the diabetic animal has type 2 diabetes.  
     
     
         64 . The method according to  claim 46 , wherein the diabetic animal is a human or a rodent.  
     
     
         65 . The method according to  claim 4 , wherein said antisense oligonucleotide comprises a first region consisting of at least 5 contiguous 2′-deoxy nucleosides flanked by second and third regions, each of said second and third regions independently consisting of at least one 2′-O-methoxyethyl nucleoside, and wherein the internucleoside linkages of the first region are phosphorothioate linkages and the internucleoside linkages of the second and third regions are phosphodiester linkages.  
     
     
         66 . The method according to  claim 24 , wherein said antisense oligonucleotide comprises a first region consisting of at least 5 contiguous 2′-deoxy nucleosides flanked by second and third regions, each of said second and third regions independently consisting of at least one 2′-O-methoxyethyl nucleoside, and wherein the internucleoside linkages of the first region are phosphorothioate linkages and the internucleoside linkages of the second and third regions are phosphodiester linkages.  
     
     
         67 . The method according to  claim 40 , wherein said compound is an antisense oligonucleotide, said antisense oligonucleotide comprising a first region consisting of at least 5 contiguous 2′-deoxy nucleosides flanked by second and third regions, each of said second and third regions independently consisting of at least one 2′-O-methoxyethyl nucleoside, and wherein the internucleoside linkages of the first region are phosphorothioate linkages and the internucleoside linkages of the second and third regions are phosphodiester linkages.  
     
     
         68 . The method according to  claim 41 , wherein said compound is an antisense oligonucleotide, said antisense oligonucleotide comprising a first region consisting of at least 5 contiguous 2′-deoxy nucleosides flanked by second and third regions, each of said second and third regions independently consisting of at least one 2′-O-methoxyethyl nucleoside, and wherein the internucleoside linkages of the first region are phosphorothioate linkages and the internucleoside linkages of the second and third regions are phosphodiester linkages.  
     
     
         69 . The method according to  claim 47 , wherein said antisense oligonucleotide comprises a first region consisting of at least 5 contiguous 2′-deoxy nucleosides flanked by second and third regions, each of said second and third regions independently consisting of at least one 2′-O-methoxyethyl nucleoside, and wherein the internucleoside linkages of the first region are phosphorothioate linkages and the internucleoside linkages of the second and third regions are phosphodiester linkages.