INHIBITION OF MIR-22 MIRNA BY APT-110
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.
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