Antisense modulation of acyl coa cholesterol acyltransferase-2 expression

Abstract

Antisense compounds, compositions and methods are provided for modulating the expression of acyl CoA cholesterol acyltransferase-2. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding acyl CoA cholesterol acyltransferase-2. Methods of using these compounds for modulation of acyl CoA cholesterol acyltransferase-2 expression and for treatment of diseases associated with expression of acyl CoA cholesterol acyltransferase-2 are provided.

1 . A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases of a nucleobase sequence selected from among the nucleobase sequences recited in SEQ ID NOs: 17, 18, 19, 20, 23, 24, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39. 
     
     
         2 . The compound of  claim 1 , wherein the modified oligonucleotide is a single-stranded oligonucleotide. 
     
     
         3 . The compound of  claim 2 , wherein the nucleobase sequence of the modified oligonucleotide is 100% complementary to acyl CoA cholesterol acyltransferase-2. 
     
     
         4 . The compound of  claim 2 , wherein at least one internucleoside linkage is a modified internucleoside linkage. 
     
     
         5 . The compound of  claim 4 , wherein each internucleoside linkage is a phosphorothioate internucleoside linkage. 
     
     
         6 . The compound of  claim 2 , wherein at least one nucleoside comprises a modified sugar. 
     
     
         7 . The compound of  claim 6 , wherein at least one modified sugar is a bicyclic sugar. 
     
     
         8 . The compound of  claim 6 , wherein at least one modified sugar comprises a 2′-O-methoxyethyl. 
     
     
         9 . The compound of  claim 2 , wherein at least one nucleoside comprises a modified nucleobase. 
     
     
         10 . The compound of  claim 9 , wherein the modified nucleobase is a 5-methylcytosine. 
     
     
         11 . The compound of  claim 1 , wherein the modified oligonucleotide comprises:
 a gap segment consisting of linked deoxynucleosides;   a 5′ wing segment consisting of linked nucleosides;   a 3′ wing segment consisting of linked nucleosides;   wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.   
     
     
         12 . The compound of  claim 11 , wherein the modified oligonucleotide comprises:
 a gap segment consisting of ten linked deoxynucleosides;   a 5′ wing segment consisting of five linked nucleosides;   a 3′ wing segment consisting of five linked nucleosides;   wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethyl sugar; and wherein each internucleoside linkage is a phosphorothioate linkage.   
     
     
         13 . The compound of  claim 2 , wherein the modified oligonucleotide consists of 20 linked nucleosides. 
     
     
         14 . A composition comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases of a nucleobase sequence selected from among the nucleobase sequences recited in SEQ ID NOs: 17, 18, 19, 20, 23, 24, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or a salt thereof and a pharmaceutically acceptable carrier or diluent. 
     
     
         15 . The composition of  claim 14 , wherein the modified oligonucleotide is a single-stranded oligonucleotide. 
     
     
         16 . The composition of  claim 14 , wherein the modified oligonucleotide consists of 20 linked nucleosides. 
     
     
         17 . A method comprising administering to an animal a compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases of a nucleobase sequence selected from among the nucleobase sequences recited in SEQ ID NOs: 17, 18, 19, 20, 23, 24, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39. 
     
     
         18 . The method of  claim 17 , wherein the animal is a human. 
     
     
         19 . The method of  claim 18 , wherein administering the compound slows the progression of a disease or condition is associated with the expression of acyl CoA cholesterol acyltranserase-2. 
     
     
         20 . The method of  claim 19 , wherein the disease or condition is hypercholesterolemia, cardiovascular disease, atherosclerosis, abnormal cholesterol metabolism or abnormal lipid metabolism. 
     
     
         21 . The method of  claim 18 , comprising co-administering the compound and an anti-obesity agent, glucose-lowering agent, and/or cholesterol-lowering agent. 
     
     
         22 . The method of  claim 21 , wherein the compound and an anti-obesity agent, glucose-lowering agent, and/or cholesterol-lowering agent are administered concomitantly. 
     
     
         23 . The method of  claim 18 , wherein the administering is topical, by inhalation or insufflation of powders or aerosols, intratracheal, intranasal, epidermal, transdermal, oral or parenteral. 
     
     
         24 . The method of  claim 23 , wherein parenteral administration comprise subcutaneous or intravenous administration. 
     
     
         25 . A method comprising identifying a human with hypercholesterolemia, cardiovascular disease, atherosclerosis, abnormal cholesterol metabolism or abnormal lipid metabolism and administering to a human a therapeutically effective amount of a composition comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases of a nucleobase sequence selected from among the nucleobase sequences recited in SEQ ID NOs: 17, 18, 19, 20, 23, 24, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39. 
     
     
         26 . The method of  claim 25 , wherein the wherein administering results in a reduction of hypercholesterolemia, cardiovascular disease, atherosclerosis, abnormal cholesterol metabolism or abnormal lipid metabolism. 
     
     
         27 . The method of  claim 25 , comprising co-administering the compound and a anti-obesity agent, glucose-lowering agent, and/or cholesterol-lowering agent. 
     
     
         28 . The method of  claim 23 , wherein the compound and an anti-obesity agent, glucose-lowering agent, and/or cholesterol-lowering agent are administered concomitantly. 
     
     
         29 . A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases, complementary to an equal-length portion of nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         30 . The compound of  claim 29 , wherein the modified oligonucleotide is at least 95% complementary to SEQ ID NO: 3. 
     
     
         31 . The compound of  claim 30 , wherein the modified oligonucleotide is 100% complementary to SEQ ID NO: 3. 
     
     
         32 . The compound of  claim 29 , wherein the modified oligonucleotide hybridizes exclusively within nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         33 . The compound of  claim 30 , wherein the modified oligonucleotide hybridizes exclusively within nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         34 . The compound of  claim 31 , wherein the modified oligonucleotide hybridizes exclusively within nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         35 . A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases fully complementary to an equal length portion of nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         36 . The compound of  claim 35 , wherein the modified oligonucleotide is at least 95% complementary to SEQ ID NO: 3. 
     
     
         37 . The compound of  claim 36 , wherein the modified oligonucleotide is 100% complementary to SEQ ID NO: 3. 
     
     
         38 . The compound of  claim 35 , wherein the modified oligonucleotide hybridizes exclusively within nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         39 . The compound of  claim 36 , wherein the modified oligonucleotide hybridizes exclusively within nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         40 . The compound of  claim 37 , wherein the modified oligonucleotide hybridizes exclusively within nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         41 . The compound of  claim 29 , consisting of a single-stranded modified oligonucleotide. 
     
     
         42 . The compound of  claim 41 , wherein at least one internucleoside linkage is a phosphorothioate internucleoside linkage. 
     
     
         43 . The compound of  claim 29 , wherein each internucleoside linkage is a phosphorothioate internucleoside linkage. 
     
     
         44 . The compound of  claim 43 , wherein at least one nucleoside comprises a modified sugar. 
     
     
         45 . The compound of  claim 44 , wherein at least one modified sugar is a bicyclic sugar. 
     
     
         46 . The compound of  claim 45 , wherein at least one modified sugar comprises a 2′-O-methoxyethyl. 
     
     
         47 . The compound of  claim 43 , wherein at least one nucleoside comprises a modified nucleobase. 
     
     
         48 . The compound of  claim 47 , wherein the modified nucleobase is a 5-methylcytosine. 
     
     
         49 . The compound of  claim 29 , wherein the modified oligonucleotide comprises:
 a gap segment consisting of linked deoxynucleosides;   a 5′ wing segment consisting of linked nucleosides;   a 3′ wing segment consisting of linked nucleosides;   wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.   
     
     
         50 . The compound of  claim 35 , wherein the modified oligonucleotide comprises:
 a gap segment consisting of ten linked deoxynucleosides;   a 5′ wing segment consisting of five linked nucleosides;   a 3′ wing segment consisting of five linked nucleosides;   wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethyl sugar; and wherein each internucleoside linkage is a phosphorothioate linkage.   
     
     
         51 . The compound of  claim 30 , wherein the modified oligonucleotide consists of 20 linked nucleosides. 
     
     
         52 . A method comprising administering to an animal a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to an equal-length portion of nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         53 . The method of  claim 52 , wherein the animal is a human. 
     
     
         54 . The method of  claim 52 , wherein administering the compound slows progression of hypercholesterolemia, cardiovascular disease, or atherosclerosis. 
     
     
         55 . The method of  claim 54 , wherein administering results in a reduction of abnormal cholesterol metabolism or abnormal lipid metabolism or combination thereof. 
     
     
         56 . The method of  claim 52 , comprising co-administering the compound and an anti-obesity agent, glucose-lowering agent, and/or cholesterol-lowering agent. 
     
     
         57 . The method of  claim 56 , wherein the anti-obesity agent, glucose-lowering agent, and/or cholesterol-lowering agent is a therapeutic lifestyle change. 
     
     
         58 . The method of  claim 52 , wherein the administering is topical, by inhalation or insufflation of powders or aerosols, intratracheal, intranasal, epidermal, transdermal, oral or parenteral. 
     
     
         59 . The method of  claim 58 , wherein the parenteral administration comprise subcutaneous or intravenous administration. 
     
     
         60 . A method comprising administering to a human with abnormal cholesterol metabolism, abnormal lipid metabolism, hypercholesterolemia, cardiovascular disease, or atherosclerosis syndrome a therapeutically effective amount of a composition comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to an equal-length portion of nucleotides 18-27, 61-100, 81-130, 111-130, 281-300, 431-450, 751-790, 881-900, 941-960, 1021-1040, 1081-1100, 1181-1200, 1221-1240, 1271-1290, 1401-1420, 1451-1470, or 1541-1560 of SEQ ID NO: 3, and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 3. 
     
     
         61 . The method of  claim 60 , wherein administering the compound slows progression hypercholesterolemia, cardiovascular disease, or atherosclerosis. 
     
     
         62 . The method of  claim 61 , wherein administering results in a reduction of abnormal cholesterol metabolism or abnormal lipid metabolism or combination thereof. 
     
     
         63 . The method of  claim 60 , comprising co-administering the compound and an anti-obesity agent, glucose-lowering agent, and/or cholesterol-lowering agent. 
     
     
         64 . The method of  claim 61 , wherein the anti-obesity agent, glucose-lowering agent, and/or cholesterol-lowering agent is a therapeutic lifestyle change. 
     
     
         65 . The method of  claim 60 , wherein the administering is topical, by inhalation or insufflation of powders or aerosols, intratracheal, intranasal, epidermal, transdermal, oral or parenteral. 
     
     
         66 . The method of  claim 61 , wherein the parenteral administration comprise subcutaneous or intravenous administration. 
     
     
         67 . A method of inhibiting the expression of acyl CoA cholesterol acyltransferase-2 in human cells or tissues, comprising contacting said cells or tissues with a compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides targeted to a nucleic acid molecule encoding human acyl CoA cholesterol acyltransferase-2. 
     
     
         68 . A method of treating hypercholesterolemia, cardiovascular disease, atherosclerosis, abnormal cholesterol metabolism or abnormal lipid metabolism by administering to a human having hypercholesterolemia, cardiovascular disease, atherosclerosis, abnormal cholesterol metabolism or abnormal lipid metabolism a compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides targeted to a nucleic acid molecule encoding hypercholesterolemia, cardiovascular disease, atherosclerosis, abnormal cholesterol metabolism or abnormal lipid metabolism. 
     
     
         69 . The method of  claim 68  wherein treating results in slowed progression and/or amelioration of hypercholesterolemia, cardiovascular disease, atherosclerosis, abnormal cholesterol metabolism or abnormal lipid metabolism. 
     
     
         70 . A method of reducing hypercholesterolemia, cardiovascular disease, atherosclerosis, abnormal cholesterol metabolism, abnormal lipid metabolism or a combination thereof comprising administering a compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides targeted to a nucleic acid molecule encoding human acyl CoA cholesterol acyltransferase-2. 
     
     
         71 . The method of  claim 70 , comprising co-administering the compound and a anti-obesity agent, glucose-lowering agent, or cholesterol-lowering agent. 
     
     
         72 . The method of  claim 71 , wherein the compound and a anti-obesity agent, glucose-lowering agent, or cholesterol-lowering agent are administered concomitantly. 
     
     
         73 . The method of  claim 72 , wherein the anti-obesity agent, glucose-lowering agent, or cholesterol-lowering agent is a therapeutic lifestyle change. 
     
     
         74 . The method of  claim 67 , wherein the compound is 90% complementary to a nucleic acid sequence encoding human acyl CoA cholesterol acyltransferase-2 selected from among the nucleic acid sequences recited in SEQ ID NO: 3. 
     
     
         75 . The method of  claim 67 , wherein the compound is 95% complementary to a nucleic acid sequence encoding human acyl CoA cholesterol acyltransferase-2 selected from among the nucleic acid sequences recited in SEQ ID NO: 3. 
     
     
         76 . The method of  claim 67 , wherein the compound is 100% complementary to a nucleic acid sequence encoding human acyl CoA cholesterol acyltransferase-2 selected from among the nucleic acid sequences recited in SEQ ID NO: 3. 
     
     
         77 . The method of  claim 67 , wherein the compound consists of a single-stranded modified oligonucleotide. 
     
     
         78 . The method of  claim 67 , wherein the compound comprises at least one phosphorothioate internucleoside linkage. 
     
     
         79 . The method of  claim 78 , wherein each internucleoside linkage in the compound is a phosphorothioate internucleoside linkage. 
     
     
         80 . The method of  claim 67 , wherein the compound comprises a modified sugar. 
     
     
         81 . The method of  claim 78 , wherein the modified sugar is a bicyclic sugar. 
     
     
         82 . The method of  claim 78 , wherein the modified sugar is a 2′-O-methoxyethyl. 
     
     
         83 . The method of  claim 67 , wherein at least one nucleoside of the compound comprises a modified nucleobase. 
     
     
         84 . The method of  claim 83 , wherein the modified nucleobase is a 5-methylcytosine. 
     
     
         85 . The method of  claim 67 , wherein the modified oligonucleotide comprises:
 a gap segment consisting of linked deoxynucleosides;   a 5′ wing segment consisting of linked nucleosides;   a 3′ wing segment consisting of linked nucleosides;   wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.   
     
     
         86 . The method of  claim 67 , wherein the modified oligonucleotide comprises:
 a gap segment consisting of ten linked deoxynucleosides;   a 5′ wing segment consisting of five linked nucleosides;   a 3′ wing segment consisting of five linked nucleosides;   wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethyl sugar; and wherein each internucleoside linkage is a phosphorothioate linkage.   
     
     
         87 . The method of  claim 67 , wherein the modified oligonucleotide consists of 20 linked nucleosides. 
     
     
         88 . The method of  claim 67 , wherein the administering is topical, by inhalation or insufflation of powders or aerosols, intratracheal, intranasal, epidermal, transdermal, oral or parenteral. 
     
     
         89 . The method of  claim 67 , wherein the paternteral administration comprise subcutaneous or intravenous administration.