US2019127736A1PendingUtilityA1

Inhibition of mir-22 mirna by apt-110

Assignee: APTAMIR THERAPEUTICS INCPriority: Apr 29, 2016Filed: Apr 28, 2017Published: May 2, 2019
Est. expiryApr 29, 2036(~9.7 yrs left)· nominal 20-yr term from priority
Inventors:Marc Thibonnier
A61K 47/14A61K 31/202C12N 2310/113C12N 2310/3231A61K 47/66A61K 31/7125C12N 2310/321A61P 3/04A61K 47/6917C12N 15/113C12N 2310/3341C12N 2310/346C12N 2310/315A61K 9/127
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Claims

Abstract

Increase of energy expenditure as an effective treatment of obesity and related disorders is a target for drug research and development. A 15% increase of energy expenditure is believed to be sufficient to achieve significant weight and fat mass reduction while providing meaningful improvement of metabolic parameters. Disclosed herein is a method for pharmacological inhibition of miR-22-3p, which represents a new therapeutic approach for treating human obesity, diabetes, and hypercholesterolemia.

Claims

exact text as granted — not AI-modified
1 . A composition comprising a compound of the formula 
       
         
           
           
               
               
           
         
         wherein X is OH or a sodium or protonated phorphorothioate 5′-hydroxy nucleotide;
 R 1  is cytosine, 5-methyl cytosine, guanine, or thymine; 
 R 2 , R 3 , R 4 , R 7 , and R 13  are each independently cytosine, 5-methyl cytosine, or thymine; 
 R 5 , R 6 , and R 10  are each independently adenine, 5-methyl cytosine, or cytosine; 
 R 8  is guanine or thymine, 
 R 9  and R 12  are each independently cytosine, 5-methyl cytosine, or guanine; 
 R 11  is adenine or guanine; 
 R 12  is cytosine, 5-methyl cytosine, or thymine; and 
 Y 1 -Y 10  are H and Z 1 -Z 10  each independently H or OMe, or 
 the respective Y and Z groups on a sugar moiety join to form an internal ether where Y is methylene and Z is an oxygen atom. 
 
       
     
     
         2 . A mir-22 miRNA antagonist of the formula 
       
         
           
           
               
               
           
         
         wherein X is OH or a sodium or protonated phorphorothioate 5′-hydroxy nucleotide;
 R 1  is cytosine, 5-methyl cytosine, guanine, or thymine; 
 R 2 , R 3 , R 4 , R 7 , and R 13  are each independently cytosine, 5-methyl cytosine, or thymine; 
 R 5 , R 6 , and R 10  are each independently adenine, 5-methyl cytosine, or cytosine; 
 R 8  is guanine or thymine, 
 R 9  and R 12  are each independently cytosine, 5-methyl cytosine, or guanine; 
 R 11  is adenine or guanine; 
 R 12  is cytosine, 5-methyl cytosine, or thymine; and 
 Y 1 -Y 10  are H and Z 1 -Z 10  each independently H or OMe, or 
 the respective Y and Z groups on a sugar moiety join to form an internal ether where Y is methylene and Z is an oxygen atom. 
 
       
     
     
         3 . The mir-22 miRNA antagonist of  claim 2 , wherein the mir-22 miRNA is the mature mir-22-3p nucleotide. 
     
     
         4 . A method of increasing thermogenesis in a subject, wherein increasing thermogenesis comprises administering to the subject the mir-22 antagonist of  claim 2 . 
     
     
         5 . A molecule comprising a therapeutic agent conjugated to a fatty acid. 
     
     
         6 . The molecule of  claim 5 , wherein the fatty acid is a C10-35 chain fatty acid. 
     
     
         7 . The molecule of  claim 5  or  6 , wherein the fatty acid is selected from decanoic acid, dodecanoic acid, oleic acid, stearic acid, docosanoic acid, and dotriacontahexaenoic acid. 
     
     
         8 . The molecule of any one of  claims 5 - 7 , wherein the therapeutic agent is a nucleic acid, a polypeptide, or a small molecule. 
     
     
         9 . The molecule of  claim 8 , wherein the therapeutic agent is a nucleic acid. 
     
     
         10 . The molecule of  claim 9 , wherein the nucleic acid is a miRNA. 
     
     
         11 . The molecule of  claim 9 , wherein the miRNA is a compound of the formula 
       
         
           
           
               
               
           
         
         wherein X is OH or a sodium or protonated phorphorothioate 5′-hydroxy nucleotide;
 R 1  is cytosine, 5-methyl cytosine, guanine, or thymine; 
 R 2 , R 3 , R 4 , R 7 , and R 13  are each independently cytosine, 5-methyl cytosine, or thymine; 
 R 5 , R 6 , and R 10  are each independently adenine, 5-methyl cytosine, or cytosine; 
 R 8  is guanine or thymine, 
 R 9  and R 12  are each independently cytosine, 5-methyl cytosine, or guanine; 
 R 11  is adenine or guanine; 
 R 12  is cytosine, 5-methyl cytosine, or thymine; and 
 Y 1 -Y 10  are H and Z 1 -Z 10  each independently H or OMe, or 
 the respective Y and Z groups on a sugar moiety join to form an internal ether where Y is methylene and Z is an oxygen atom. 
 
       
     
     
         12 . A liposome comprising phospholipids and cholesterol. 
     
     
         13 . The liposome of  claim 12 , wherein the weight ratio of phospholipids to cholesterol is 80:20% weight to weight ratio. 
     
     
         14 . The liposome of  claim 12  or  13 , wherein the phospholipids are selected from sphingomyelin, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), or both. 
     
     
         15 . The liposome of  claim 14 , wherein the liposome comprises both sphingomyelin and DMPC. 
     
     
         16 . The liposome of  claim 15 , wherein the weight ratio of sphingomyelin, DMPC, and cholesterol is 40:40:20% weight to weight ratio. 
     
     
         17 . The liposome of any one of  claims 12 - 16 , wherein the liposome further comprises a therapeutic agent. 
     
     
         18 . The liposome of  claim 17 , wherein the therapeutic agent is a polypeptide, a nucleic acid, or a small molecule. 
     
     
         19 . The liposome of  claim 18 , wherein the therapeutic agent comprises the molecule of any one of  claims 5 - 11 . 
     
     
         20 . The liposome of any one of  claims 12 - 19  wherein the diameter is 100-200 nm. 
     
     
         21 . The liposome of any one of  claims 12 - 20 , wherein the liposome comprises a targeting element. 
     
     
         22 . The liposome of  claim 21 , wherein the targeting element is a TSP-1 polypeptide. 
     
     
         23 . The liposome of  claim 22 , wherein the TSP-1 polypeptide comprises the sequence: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 1) 
                 
                     
                   GVITRIR. 
                 
             
                
                
               
            
           
         
       
     
     
         24 . The liposome of  claim 21 , wherein the targeting element is an Hexarelin polypeptide. 
     
     
         25 . The liposome of  claim 24 , wherein the Hexarelin polypeptice comprises the sequence 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 2) 
                 
                     
                   HWAWFL. 
                 
             
                
                
               
            
           
         
       
     
     
         26 . The liposome of  claim 21 , wherein the targeting element is Prohibitin polypeptide. 
     
     
         27 . The liposome of  claim 26 , wherein the Prohibitin polypeptice comprises the sequence 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 3) 
                 
                     
                   CKGGRAKDC. 
                 
             
                
                
               
            
           
         
       
     
     
         28 . The liposome of any one of  claims 17 - 23 , wherein the therapeutic agent is a compound of the formula: 
       
         
           
           
               
               
           
         
         wherein X is OH or a sodium or protonated phorphorothioate 5′-hydroxy nucleotide;
 R 1  is cytosine, 5-methyl cytosine, guanine, or thymine; 
 R 2 , R 3 , R 4 , R 7 , and R 13  are each independently cytosine, 5-methyl cytosine, or thymine; 
 R 5 , R 6 , and R 10  are each independently adenine, 5-methyl cytosine, or cytosine; 
 R 8  is guanine or thymine, 
 R 9  and R 12  are each independently cytosine, 5-methyl cytosine, or guanine; 
 R 11  is adenine or guanine; 
 R 12  is cytosine, 5-methyl cytosine, or thymine; and 
 Y 1 -Y 10  are H and Z 1 -Z 10  each independently H or OMe, or 
 the respective Y and Z groups on a sugar moiety join to form an internal ether where Y is methylene and Z is an oxygen atom. 
 
       
     
     
         29 . A method for delivering a therapeutic agent to an adipocyte comprising administering the molecule of any one of  claims 5 - 11  or the liposome of any one of  claims 12 - 28  to the adipocyte. 
     
     
         30 . The method of  claim 29 , wherein the adipocyte is in vivo. 
     
     
         31 . The method of  claim 29 , wherein the biodistribution of the therapeutic agent is at least 50% in adipocytes. 
     
     
         32 . A method of inhibiting mir-22 in a cell comprising administering to the cell the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         33 . The method of  claim 30 , wherein the cell is an adipocyte, pre-adipocyte, fibroblast, or vascular endothelial cell. 
     
     
         34 . The method of  claim 33 , wherein the cell is an adipose tissue cell. 
     
     
         35 . The method of  claim 34 , wherein the adipose tissue cell is subcutaneous white adipose cell or brown adipose tissue cell. 
     
     
         36 . A method for upregulating the histone demethylase activity of KDM3A or KDM6B in a cell, comprising administering to the cell the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         37 . The method of  claim 36 , wherein upregulation of KDM3A increases histone demethylation in the PPARA gene region. 
     
     
         38 . The method of  claim 37 , wherein increasing histone demethylation in the PPARA gene region upregulates expression of PPARA. 
     
     
         39 . The method of  claim 38 , wherein upregulating expression of PPARA activates lipid catabolism and thermogenesis. 
     
     
         40 . The method of  claim 36 , wherein upregulation of KDM3A increases histone demethylation in the UCP1 gene region. 
     
     
         41 . The method of  claim 40 , wherein increasing histone demethylation in the UCP1 gene region upregulates expression of UCP1. 
     
     
         42 . The method of  claim 41 , wherein upregulating expression of UCP1 increases mitochondrial proton leak. 
     
     
         43 . The method of  claim 42 , wherein increasing mitochondrial proton leak increases cellular thermogenesis. 
     
     
         44 . The method of  claim 42 , wherein increasing mitochondrial proton leak increases cellular caloric expenditure. 
     
     
         45 . A method of affecting weight loss in a subject, wherein affecting weight loss comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         46 . A method of increasing caloric expenditure in a subject, wherein increasing caloric expenditure comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         47 . A method of decreasing total fat mass in a subject, wherein decreasing total fat mass comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         48 . A method of decreasing blood glucose levels in a subject, wherein decreasing blood glucose levels comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         49 . A method of decreasing blood insulin levels in a subject, wherein decreasing blood insulin levels comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         50 . A method of decreasing blood leptin levels in a subject, wherein decreasing blood leptin levels comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         51 . A method of maintaining insulin sensitivity in a subject, wherein maintaining insulin sensitivity comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         52 . A method for converting white adipocytes to brown adipocytes, wherein converting white adipocytes to brown adipocytes comprises administering to the subject the mir-22 antagonist of  claim 1  or  2  or the molecule or liposome of  claim 22 . 
     
     
         53 . A method for increasing lipolysis in a subject, wherein increasing lipolysis comprises administering to the subject the mir-22 antagonist of  claim 1  or  2  or the molecule or liposome of  claim 22 . 
     
     
         54 . A method for increasing beta-oxidation of fatty acids in a subject, wherein increasing beta-oxidation of fatty acids comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         55 . A method of increasing thermogenesis in a subject, wherein increasing thermogenesis comprises administering to the subject the mir-22 antagonist of  claim 2 , molecule of  claim 5 , or liposome of  claim 28 . 
     
     
         56 . The composition of  claim 1 , wherein the structure is 
       
         
           
           
               
               
           
         
       
     
     
         57 . The composition of  claim 1 , wherein the structure is 
       
         
           
           
               
               
           
         
       
     
     
         58 . The composition of  claim 1 , wherein the structure is 
       
         
           
           
               
               
           
         
       
     
     
         59 . A composition comprising a compound of the formula 
       
         
           
           
               
               
           
         
       
     
     
         60 . The miRNA of  claim 11 , wherein the miRNA is 
       
         
           
           
               
               
           
         
       
     
     
         61 . The miRNA of  claim 11 , wherein the miRNA is 
       
         
           
           
               
               
           
         
       
     
     
         62 . The miRNA of  claim 11 , wherein the miRNA is 
       
         
           
           
               
               
           
         
       
     
     
         63 . The therapeutic agent of  claim 28 , wherein the therapeutic agent is 
       
         
           
           
               
               
           
         
       
     
     
         64 . The therapeutic agent of  claim 28 , wherein the therapeutic agent is 
       
         
           
           
               
               
           
         
       
     
     
         65 . The therapeutic agent of  claim 28 , wherein the therapeutic agent is 
       
         
           
           
               
               
           
         
       
     
     
         66 . A molecule comprising a a fatty acid conjugated to a therapeutic agent of the structure 
       
         
           
           
               
               
           
         
       
     
     
         67 . A liposome comprising phospholipids, cholesterol, and a therapeutic agent of the structure

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