Microrna inhibitors comprising locked nucleotides

Abstract

The invention provides chemically modified oligonucleotides capable of inhibiting the expression (e.g., abundance) of miR-208 family miRNAs, including miR-208a, miR-208b, and/or miR-499. The invention provides in some embodiments, oligonucleotides capable of inhibiting, in a specific fashion, the expression or abundance of each of miR-208a, miR-208b, and miR-499. The invention further provides pharmaceutical compositions comprising the oligonucleotides, and methods of treating patients having conditions or disorders relating to or involving a miR-208 family miRNA, such as a cardiovascular condition. In various embodiments, the oligonucleotides provide advantages in one or more of potency, efficiency of delivery, target specificity, toxicity, and/or stability.

1 . An oligonucleotide comprising a nucleotide sequence that is substantially complementary to a nucleotide sequence of human miR-208a or miR-208b, and having a mix of locked and non-locked nucleotides,
 wherein the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces miR-208a, miR-208b, and/or miR-499 activity at an oligonucleotide concentration of about 50 nM or less in an in vitro luciferase assay, or at a dose of 25 mg/kg or less in a mouse model.   
     
     
         2 - 47 . (canceled) 
     
     
         48 . A method of preventing or treating pathologic cardiac hypertrophy in a subject associated with or mediated by miR-208a, miR-208(b), and/or miR-499, comprising administering to the subject a pharmaceutical composition comprising an effective amount of an oligonucleotide comprising a nucleotide sequence that is substantially complementary to a nucleotide sequence of human miR-208a or miR-208b, and having a mix of locked and non-locked nucleotides,
 wherein the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces miR-208a, miR-208b, and/or miR-499 activity at an oligonucleotide concentration of about 50 nM or less in an in vitro luciferase assay, or at a dose of 25 mg/kg or less in a mouse model.   
     
     
         49 . A method of preventing or treating myocardial infarction in a subject associated with or mediated by miR-208a, miR-208(b), and/or miR-499, comprising administering to the subject a pharmaceutical composition comprising an effective amount of an oligonucleotide comprising a nucleotide sequence that is substantially complementary to a nucleotide sequence of human miR-208a or miR-208b, and having a mix of locked and non-locked nucleotides,
 wherein the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces miR-208a, miR-208b, and/or miR-499 activity at an oligonucleotide concentration of about 50 nM or less in an in vitro luciferase assay, or at a dose of 25 mg/kg or less in a mouse model.   
     
     
         50 . A method of preventing or treating heart failure in a subject associated with or mediated by miR-208a, miR-208(b), and/or miR-499, comprising administering to the subject a pharmaceutical composition comprising an effective amount of an oligonucleotide comprising a nucleotide sequence that is substantially complementary to a nucleotide sequence of human miR-208a or miR-208b, and having a mix of locked and non-locked nucleotides,
 wherein the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces miR-208a, miR-208b, and/or miR-499 activity at an oligonucleotide concentration of about 50 nM or less in an in vitro luciferase assay, or at a dose of 25 mg/kg or less in a mouse model.   
     
     
         51 . A method of preventing or treating vascular damage in a subject associated with or mediated by miR-208a, miR-208(b), and/or miR-499, comprising administering to the subject a pharmaceutical composition comprising an effective amount of an oligonucleotide comprising a nucleotide sequence that is substantially complementary to a nucleotide sequence of human miR-208a or miR-208b, and having a mix of locked and non-locked nucleotides,
 wherein the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces miR-208a, miR-208b, and/or miR-499 activity at an oligonucleotide concentration of about 50 nM or less in an in vitro luciferase assay, or at a dose of 25 mg/kg or less in a mouse model.   
     
     
         52 . A method of preventing or treating restenosis in a subject associated with or mediated by miR-208a, miR-208(b), and/or miR-499, comprising administering to the subject a pharmaceutical composition comprising an effective amount of an oligonucleotide comprising a nucleotide sequence that is substantially complementary to a nucleotide sequence of human miR-208a or miR-208b, and having a mix of locked and non-locked nucleotides,
 wherein the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces miR-208a, miR-208b, and/or miR-499 activity at an oligonucleotide concentration of about 50 nM or less in an in vitro luciferase assay, or at a dose of 25 mg/kg or less in a mouse model.   
     
     
         53 . A method of preventing or treating pathologic cardiac fibrosis in a subject associated with or mediated by miR-208a, miR-208(b), and/or miR-499, comprising administering to the subject a pharmaceutical composition comprising an effective amount of an oligonucleotide comprising a nucleotide sequence that is substantially complementary to a nucleotide sequence of human miR-208a or miR-208b, and having a mix of locked and non-locked nucleotides,
 wherein the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces miR-208a, miR-208b, and/or miR-499 activity at an oligonucleotide concentration of about 50 nM or less in an in vitro luciferase assay, or at a dose of 25 mg/kg or less in a mouse model.