Modulation of glucagon receptor expression

Abstract

Compounds, compositions and methods are provided for modulating the expression of glucagon receptor. The compositions comprise oligonucleotides, targeted to nucleic acid encoding glucagon receptor. Methods of using these compounds for modulation of glucagon receptor expression and for diagnosis and treatment of disease associated with expression of glucagon receptor are provided.

1 . An antisense compound 12 to 50 nucleobases in length comprising at least one of a modified internucleoside linkage, a high affinity modified sugar or a modified nucleobase, wherein the compound targets and specifically hybridizes with at least 90% complementarity within nucleotides 532 to 558 of a nucleic acid molecule encoding a human glucagon receptor having SEQ ID NO:4. 
     
     
         2 . The compound of  claim 1  which is 15 to 30 nucleobases in length. 
     
     
         3 . The compound of  claim 1  which is 20 nucleobases in length. 
     
     
         4 . The compound of  claim 1  having at least 95%. complementarity with a nucleic acid molecule encoding human glucagon receptor having SEQ ID NO: 4. 
     
     
         5 . The compound of  claim 1  having at least one 2′-O-methoxyethyl sugar moiety. 
     
     
         6 . The compound of  claim 1  having at least one phosphorothioate internucleoside linkage. 
     
     
         7 . The compound of  claim 1  having at least one 5-methylcytosine. 
     
     
         8 . The compound of  claim 1  that is a pharmaceutically acceptable salt. 
     
     
         9 . A pharmaceutical composition comprising the compound of  claim 1  and a pharmaceutically acceptable diluent or carrier. 
     
     
         10 . The compound of  claim 1  comprising at least one high affinity modified sugar moiety selected from the group consisting of a 2′-O-(2-methoxyethyl), a 2′-O-methyl or a locked nucleic acid. 
     
     
         11 . The compound of  claim 1  wherein the compound is chimeric, comprising deoxynucleotides in a gap region, at least one high affinity modified sugar in each of a first wing region, and a second wing region, said wing regions flanking the gap region on the 5′ end and the 3′ end, respectively, and at least one phosphorothioate modified internucleoside linkage. 
     
     
         12 . The compound of  claim 11  wherein the gap segment is ten deoxynucleotides in length, the first and second wing segments are each five nucleotides in length and comprise five 2′-O-(2-methoxyethyl) nucleotides, and each internucleoside linkage in the chimeric oligonucleotide is a phosphorothioate. 
     
     
         13 . The compound of  claim 1  wherein at least a portion of said compound hybridizes with RNA to form an oligonucleotide-RNA duplex. 
     
     
         14 . The compound of  claim 11  further comprising at least one 5-methylcytosine modified nucleobase. 
     
     
         15 . The compound of  claim 1  comprising a pharmaceutically acceptable salt. 
     
     
         16 . The compound of  claim 1 , wherein the compound targets and specifically hybridizes within nucleotides 532 to 558. 
     
     
         17 . A method of inhibiting the expression of human glucagon receptor in cells or tissues comprising contacting said cells or tissues with an antisense compound 12 to 50 nucleobases in length comprising at least one of a modified internucleoside linkage, a high affinity modified sugar or a modified nucleobase, wherein the compound is targeted to a nucleic acid molecule encoding human glucagon receptor and wherein the compound specifically hybridizes with at least 90% complementarity with nucleotides 532 to 558 of a nucleic acid molecule having the sequence SEQ ID NO:4 and inhibits the expression of the human glucagon receptor. 
     
     
         18 . A method of treating or delaying the onset of a disease or condition associated with glucagon receptor in a human comprising administering to said human a therapeutically or prophylactically effective amount of an antisense compound 12 to 50 nucleobases in length comprising at least one of a modified internucleoside linkage, a high affinity modified sugar or a modified nucleobase, wherein the compound is targeted to a nucleic acid molecule encoding human glucagon receptor and wherein the compound specifically hybridizes with at least 90% complementarity with nucleotides 532 to 558 of a nucleic molecule having the sequence SEQ ID NO:4, and inhibits the expression of the human glucagon receptor, thereby treating or delaying the onset of the disease or condition. 
     
     
         19 . The method of  claim 18 , wherein the disease or condition is a metabolic disease or condition. 
     
     
         20 . The method of  claim 19 , wherein the metabolic disease or condition is diabetes, obesity, hyperglycemia, primary hyperglucagonemia, insulin deficiency or insulin resistance. 
     
     
         21 . The method of  claim 20  wherein the diabetes is Type 2 diabetes. 
     
     
         22 . The method of  claim 19 , wherein the metabolic disease or condition is associated with elevated blood glucose levels, elevated blood triglyceride levels or elevated blood cholesterol levels. 
     
     
         23 . A method of decreasing blood glucose levels in a human comprising administering to said human a therapeutically or prophylactically effective amount of an antisense compound 12 to 50 nucleobases in length comprising at least one of a modified internucleoside linkage, a high affinity modified sugar or a modified nucleobase, wherein the compound is targeted to a nucleic acid molecule encoding human glucagon receptor and wherein the compound specifically hybridizes with at least 90% complementarity within nucleotides 532 to 558 of a nucleic acid molecule having the sequence SEQ ID NO:4 and inhibits the expression of the human glucagon receptor. 
     
     
         24 . The method of  claim 23  wherein the blood glucose levels are plasma glucose levels. 
     
     
         25 . The method of  claim 23  wherein the human suffers from diabetes or obesity. 
     
     
         26 . A method of preventing or delaying the onset of an increase in blood glucose levels in a human comprising administering to said human a therapeutically or prophylactically effective amount of an antisense compound 12 to 50 nucleobases in length comprising at least one of a modified internucleoside linkage, a high affinity modified sugar or a modified nucleobase, wherein the compound is targeted to a nucleic acid molecule encoding human glucagon receptor and wherein the compound specifically hybridizes with at least 90% complementarity within nucleotides 532 to 558 of a nucleic acid molecule having the sequence SEQ ID NO:4 and inhibits the expression of the human glucagon receptor. 
     
     
         27 . The method of  claim 26  wherein the human suffers from diabetes, obesity or insulin resistance. 
     
     
         28 . The method of  claim 26  wherein the blood glucose levels are plasma glucose levels. 
     
     
         29 . The method of  claim 17 , wherein the compound is at least 95% complementary to SEQ ID NO: 4. 
     
     
         30 . The method of  claim 17 , wherein the compound is 15 to 30 nucleobases in length. 
     
     
         31 . The method of  claim 17 , wherein the compound is 20 nucleobases in length. 
     
     
         32 . The method of  claim 17 , wherein the compound comprises an oligonucleotide. 
     
     
         33 . The method of  claim 32 , wherein the oligonucleotide is chimeric. 
     
     
         34 . The method of  claim 17 , wherein the compound comprises at least one chemical modification. 
     
     
         35 . The method of  claim 34 , wherein the compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase. 
     
     
         36 . The method of  claim 35 , wherein the compound comprises at least one 2′-O-(2-methoxyethyl) sugar moiety. 
     
     
         37 . The method of  claim 35 , wherein the compound comprises at least one phosphorothioate internucleoside linkage. 
     
     
         38 . The method of  claim 35 , wherein the compound comprises at least one 5-methylcytosine. 
     
     
         39 . A method of  claim 33 , wherein every internucleoside linkage of the oligonucleotide is a phosphorothioate linkage, nucleobases 1-5 and 16-20 of the oligonucleotide comprise a 2′-O-(2-methoxyethyl) modification and every cytidine residue comprises a 5-methyl modification. 
     
     
         40 . The method of  claim 17 , wherein the compound is a sodium salt. 
     
     
         41 . The method of  claim 17 , wherein the compound is administered as a composition comprising a pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         42 . The method of  claim 41 , wherein the composition farther comprises a colloidal dispersion system. 
     
     
         43 . The method of  claim 18 , wherein administering comprises delivering a plurality of doses. 
     
     
         44 . The method of  claim 43 , wherein each dose of said plurality of doses comprises from about 1 mg about 20 mg of the compound per kg of body weight. 
     
     
         45 . The method of  claim 17 , wherein the compound is 100% complementary to SEQ ID NO:4. 
     
     
         46 . The method of  claim 17 , wherein the compound targets and specifically hybridizes within nucleotides 532 to 558.