WO2013192486A1 - Inhibitors of the mir-15 family of micro-rnas - Google Patents
Inhibitors of the mir-15 family of micro-rnas Download PDFInfo
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- WO2013192486A1 WO2013192486A1 PCT/US2013/046960 US2013046960W WO2013192486A1 WO 2013192486 A1 WO2013192486 A1 WO 2013192486A1 US 2013046960 W US2013046960 W US 2013046960W WO 2013192486 A1 WO2013192486 A1 WO 2013192486A1
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- oligonucleotide
- mir
- nucleotides
- seq
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Classifications
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- C12N15/09—Recombinant DNA-technology
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- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
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- A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
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- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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Definitions
- the present invention relates to chemical motifs for microRNA (miRNA or nilR) inhibitors. More particularly, the present invention relates to chemically modified oligonucleotides that target one or more miR- 15 family members and having advantages in potency, efficiency of delivery, target specificity, stability, and/or toxicity when administered to a patient.
- miRNA miRNA or nilR
- MieroRNAs have been implicated in a number of biological processes including regulation and maintenance of cardiac function (see, E. Van Rooij et a!., J. Clin, invest. 117(9): 2369-2376 (2007); K. Chien, Nature 447: 389-390 (2007)). Therefore, miRs represent a relatively new class of therapeutic targets for conditions such as cardiac hypertrophy, myocardial infarction, heart failure, vascular damage, and pathologic cardiac fibrosis, among others.
- miRs are small, non-protein coding RNAs of about 18 to about 25 nucleotides in length, and act as repressors of target mRNAs by promoting their degradation, when their sequences are perfectly complementary, or by inhibiting translation, when their sequences contain mismatches.
- the mechanism involves incorporation of the mature miRNA strand into the RNA-induced silencing complex (RISC), where it associates with its target RNAs by base-pair complementarity.
- miRNA function may be targeted therapeutically by antisense polynucleotides or by polynucleotides that mimic miRNA function ("miRNA mimetic").
- RNA-binding affinity and specificity RNA-binding affinity and specificity
- efficiency of cellular uptake RNA-binding affinity and specificity
- nuclease resistance RNA-binding affinity and specificity
- polynucleotides when polynucleotides are introduced into intact cells they are attacked and degraded by nucleases leading to a loss of activity.
- polynucleotide analogues have been prepared in an attempt to avoid their degradation, e.g. by means of 2' substitutions (B. Sproat et al, Nucleic Acids Research 17: 3373-3386, (1989)), the modifications often affect the polynucleotide's potency for its intended biological action.
- Such reduced potency may be due to an inability of the modified polynucleotide to form a stable duplex with the target RNA and/or a loss of interaction with the cellular machinery.
- Other modifications include the use of locked nucleic acid, which has the potential to improve RNA-binding affinity (see, R. Veedu et a!., RNA Biology 6(3): 321-323 (2009)).
- Oligonucleotide chemistry patterns or motifs for miRNA inhibitors have the potential to improve the delivery, stability, potency, specificity, and/or toxicity profile of the inhibitors, and such are needed for effectively targeting miRNA function in a therapeutic context.
- the present invention provides chemically modified oligonucleotides capable for reducing or inhibiting the activity of one or more mill- 15 family members including miR- 15a, miR-15b, miR-16, miR-195, miR-424, and miR-497.
- the invention further provides pharmaceutical compositions comprising the oligonucleotides, and methods of treating subjects having conditions or disorders relating to, or involving, one or more miR-15 family members.
- the disclosed oligonucleotides provide advantages in one or more of potency, efficiency of delivery, target specificity, toxicity, and/or stability.
- the invention provides a chemically modified oligonucleotide capable of reducing or inhibiting the activity of one or more mi.R-15 family members.
- the activity or potency of the oligonucleotide may be determined in vitro and/or in vivo.
- the oligonucleotide may significantly inhibit ⁇ e.g., about 50% inhibition) the activity of one or more miR-15 family members (as determined, for example, in a two-step real time PGR assay or a dual luciterase assay) at a concentration of about 50 nM or less, or in other embodiments, about 40 nM or less, about 20 nM or less, or about 10 nM or less.
- the activity of the oligonucleotide may be determined in a suitable mouse or rat model, or non-human primate model, where inhibition ⁇ e.g., by at least about 50%) of one or more miR-15 family members is observed at a dose of about 50 mg/kg or less, such as about 25 mg/kg or less, about 10 mg/kg or less, or about 5 mg kg or less. Exemplary markers for determining target de-repression are described herein.
- the oligonucleotide may be dosed subcutaneously or intravenously (and as described herein), and may be formulated in an aqueous preparation (e.g., saline).
- the oligonucleotide comprises the nucleotide sequence 5'-GTGCTGCT-3' and is substantially complementary to a nucleotide sequence of one or more miR-15 family members.
- the oligonucleotide further contains at least one locked nucleotide.
- the oligonucleotide contains at least eight locked nucleotides.
- the locked nucleotides may have a 2' to 4' bridge structure as described in WO 2012/083005, which is incorporated herein in its entirety.
- the locked nucleotide may have a 2' to 4' bridge comprising an ethylene or methylene group.
- the locked nucleotide may have a 2 " to 4' methylene bridge.
- the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces the activity of one or more miR-15 family members at an oligonucleotide concentration of about 50 nM or less in an in vitro real time PGR assay or a luciterase assay, or at a dose of about 25 mg kg or less in a suitable rodent model or non-human primate model as described herein.
- the oligonucleotide is from about 8 to about 18 nucleotides in length. In another embodiment, the oligonucleotide is from about 12 to about 17 nucleotides in length.
- the oligonucleotide is about 16 nucleotides in length.
- the oligonucleotide may comprise or consist essentially of a nucleotide sequence selected from 5 ' - ACC ATT ATGTGCTGCT-3 ' (SEQ ID NO. 1), 5 ' - ACC ATGATGTGCTGCT-
- the oligonucleotide may include nine locked nucleotides and seven non- locked nucleotides.
- the pattern of locked nucleotides may be such that at least positions 1 , 5, 8, 10, and 16 are locked nucleotides.
- the oligonucleotide is about 12 nucleotides in length.
- the oligonucleotide may comprise or consist essentially of a nucleotide sequence selected from 5 '- TTATGTGCTGCT-3 '(SEQ ID NO. 5), 5 '-TTACGTGCTGCT-3 '(SEQ ID NO. 6), 5 '-TTCTGTGCTGCT-3 '(SEQ ID NO. 7), 5 '-TTCCGTGCTGCT-3 '(SEQ ID NO. 8), 5 ' -TG ATGTGCTG CT-3 ' (SEQ ID NO. 9), 5 '-TGACGTGCTGCT-3 '(SEQ ID NO.
- the oligonucleotide may include eight locked nucleotides and four non- locked nucleotides.
- the pattern of locked nucleotides may be such that at least positions 1 , 4, 9, and 12 are locked nucleotides.
- the oligonucleotide is about 8 nucleotides in length .
- the oligonucleotide may consist essentially of, or consists of the nucleotide sequence 5 '- GTGCTGCT-3 ', where all or substantially all (e.g., at least about 6 or at least about 7) are locked nucleotides.
- the region complementary to the seed region of the one or more mill- 15 family members comprises at least about four locked nucleotides, in another embodiment, the oligonucleotide does not contain a stretch of nucleotides with more than three contiguous non-locked nucleotides and/or more than three contiguous locked nucleotides.
- the nucleotide may contain a 2' modification with respect to a 2' hydroxy!.
- the 2' modification may be independently selected from deoxy, O-alkyl, O-methyl, halo, and fluoro.
- the 2' modification may be deoxy.
- all the non-locked nucleotides in the oligonucleotide are 2' deoxy.
- the oligonucleotide may also contain one or more phosphorothioate linkages.
- the oligonucleotide may be fully phosphorothioate-linked or may contain about half or 3 ⁇ 4 phosphorothioate linkages.
- oligonucleotide inhibitors are shown in Table 1.
- the oligonucleotide may contain a 5' and or a 3' cap structure.
- the oligonucleotide may further include a pendent lipophilic group.
- the invention provides pharmaceutical compositions and formulations comprising the oligonucleotides of the invention, which may involve incorporation of the oligonucleotide within a variety of macromolecular assemblies, micelle, or liposome compositions for cellular delivery.
- the oligonucleotides are formulated for conventional intravenous, subcutaneous, or intramuscular dosing. Such formulations may be conventional aqueous preparations, such as formulation in saline.
- the compositions are suitable or formulated for intradermal, subcutaneous, intramuscular, intraperitoneal or intravenous injection, or by direct injection into target tissue (e.g., cardiac tissue).
- the invention provides a method for delivering the oligonucleotide and the pharmaceutical composition to mammalian cells either in vitro or ex vivo, e.g., for treating, ameliorating, or preventing the progression of a condition in a mammalian subject.
- the method may comprise administering the oligonucleotide or composition comprising the same to a population of target cells or a mammalian subject at a dose effective for target de-repression of Piml .
- the subject may have a condition associated with, mediated by, or resulting from, the expression of one or more miR-15 family members.
- Such conditions include, for example, cardiac hypertrophy, myocardial infarction, heart failure (e.g., congestive heart failure), ischemia, ischemia reperfusion injury, vascular damage, restenosis, pathologic cardiac fibrosis, or conditions associated with cardiac transplantation.
- heart failure e.g., congestive heart failure
- ischemia ischemia reperfusion injury
- vascular damage vascular damage
- restenosis pathologic cardiac fibrosis
- pathologic cardiac fibrosis or conditions associated with cardiac transplantation.
- FIG. 1 Abundance of miR-15 family members in cardiocytes. MicroRNA copy numbers per cell are measured by real-time PGR analysis and normalized to a commercially available standard (Ambion).
- FIG. 1 miR-15 targets in cardiac tissue.
- Real-time PGR analysis demonstrates that subcutaneous delivery of 25 mg/kg of anti-miR.15b (M-10134) induces robust changes in the expression of Bcl2L2, Birc5, Grn, and Cdc2A in mice.
- Anti-miR15b (M-10134) treatment affects target gene expression in a dose- dependent manner.
- Real-time PCR. analysis demonstrates a dose-dependent de-repression of miR-15 targets, particularly, Birc5, Grn, and Cdc2A in mice.
- M-10591 is a non- targeting control oligonucleotide.
- Figure 5 The TaqMan* MicroRNA Assay (Applied Biosystems) for quantifying inhibitor activity in vitro using real-time PCR. analysis.
- Figure 6A shows miR-15 inhibitor efficacy measured by two-step real-time PCR analysis for mi.R-15a, miR-15b, mi.R-16, and miR-195.
- Figure 6B is a chart summarizing the efficacy of each miR-15 inhibitor.
- Figure 7 The psiCHEC TM-2 construct (Promega) for quantifying inhibitor activity in vitro using the dual luciferase assay.
- Figure 8A shows miR-15 inhibitor efficacy measured by dual luciferase assay for miR-15a, miR ⁇ 15b, miR-16, and miR-195.
- Figure 8B is a chart, summarizing the efficacy of each miR-15 inhibitor.
- Figure 9A shows that a therapeutic regimen of three doses (25 mg/kg per dose) of miR-15 inhibitors induces target gene de-repression in mice. Statistically significant groups with the indicated p values are indicated with an asterisk.
- Figure 9B shows that a therapeutic regimen of three doses (25 mg kg per dose) of miR-15 inhibitors is more potent than a single dose regimen (25 mg/kg per dose).
- Figure 10. Kinetics of target gene de-repression by miR-15 inhibitors.
- Figure 10 shows that delivery of a single dose (25 mg/kg) of M- 1 .121.5 induces potent, silencing of miR-15 targets in mice as early as 24 hours post injection.
- Figure 11 Global and specific target, gene de-repression by miR-15 inhibitors.
- Figure 11 shows that M- 11214 elicits target gene de-repression that is specific to the miR-15 family (p-value: 0.01).
- the p-value for enrichment was calculated using a hypergeometric distribution function.
- Figure 12 Efficacy of a panel of miR-15 inhibitors on target gene de-repression in rats.
- Figure 12A shows subcutaneous injection of a single dose of miR-15 inhibitors (25 mg/kg) induces target gene de-repression four days after treatment. In rats, the 16-mer inhibitors appear to be more potent than the 12-mer inhibitors.
- Figure 12B compares target gene de-repression by miR-15 inhibitors in rats versus in mice at day two after treatment. The data for the graphs on the left were generated in rats, and the data for the graphs on the right were generated in mice. In general, miR-15 inhibitors appear to have a more potent effect in rats than in mice, with the exception of 12-mer inhibitors, which do not appear to be efficacious in rats.
- Figure 13 Efficacy of miR-15 inhibitors on target gene de-repression in a rat stress model.
- Baseline represents non-infarcted controls.
- IR/Saline represents saline treated controls.
- M-10591 is a non-targeting control oligonucleotide.
- Figure 14 Efficacy of miR- 15 inhibitors on the expression of inflammatory markers in a rat stress model.
- Baseline represents non-infarcted controls.
- IR/Saline represents saline treated controls.
- M-10591 is a non-targeting control oligonucleotide.
- Figure 15 Efficacy of miR- 15 inhibitors on myocardial infarction in a rat stress model.
- Saline represents saline treated controls.
- M-10591 is a non- targeting control oligonucleotide.
- Figure 16 Efficacy of miR- 15 inhibitors (M-1 121 1 and M-1 1214) on ejection fraction in a rat stress model.
- Figure 16 shows the effects of miR- 15 inhibitors on the ejection fraction in a rat model of ischemia-reperfusion injury.
- AMC represents age matched controls without injury.
- IR/Saline represents saline treated controls.
- the invention provides chemically modified oligonucleotides capable of inhibiting the expression (e.g., abundance) of miR- 15 family miR As, including miR- 15a, miR- 15b, miR- 16, miR-195, miR-424, and miR-497.
- the invention provides in some embodiments, oligonucleotides capable of inhibiting, in a specific fashion, the expression or abundance of each of miR-1 a, miR- 15b, mi R- 1 6, miR-195, miR-424, and miR-497.
- the invention further provides pharmaceutical compositions comprising the oligonucleotides, and methods of treating patients having conditions or disorders relating to or involving a miR- 15 family miRNA, such as a various cardiovascular conditions, in various embodiments, the oligonucleotides provide advantages in one or more of potency, efficiency of delivery, target specificity, toxicity, and/or stability. in one aspect, the invention provides an oligonucleotide capable of reducing the expression or abundance of miR-15 family miRNAs. The activity of the oligonucleotides may be determined in vitro and/or in vivo. For example, when inhibition of miR-15 a, miR-!
- the activity may be determined using a two-step real-time PGR assay or a dual luciferase assay as described herein.
- the oligonucleotide significantly inhibits such activity, as determmed, for example, in the dual luciferase assay, at a concentration of about 75 nM or less, or in other embodiments, about 50 nM or less, about 40 nM or less, about 20 nM or less, or about 10 nM or less.
- the oligonucleotide may have an IC50 for inhibition of miR-15 a, miR-15b, miR-16, miR-195, miR-424, and miR-497 activity of about 50 nM or less, about 40 n ⁇ 1 or less, about 30 nM or less, or about 20 nM or less, as determined in the dual luciferase assay.
- the two-step PCR analysis involves a quantitative PCR readout which can be inhibited at two steps by an oligonucleotide inhibitor: 1 ) by inhibiting the ability of the reverse transcriptase primer to elongate during cDNA synthesis; and 2) by inhibiting the ability of the PCR primer to amplify the cDNA product.
- the dual luciferase assay involves placement of the miR recognition site in the 3' UTR of a gene for a detectable protein ⁇ e.g., renilla luciferase).
- the construct is co-expressed with the target miRNA, such that inhibitor activity can be determined by change in signal.
- a second gene encoding a detectable protein ⁇ e.g., firefly luciferase can be included on the same plasmid, and the ratio of signals determined as an indication of anti-rm ' R activity .
- the activity of the oligonucleotide may be determined in a suitable mouse or rat model, such as those described herein, where inhibition ⁇ e.g., by at least about 50%) of a miR-15 family miRNA. is observed at an oligonucleotide dose of about 50 mg kg or less, about 25 mg/kg or less, such as about 10 mg/kg or less or about 5 mg/kg or less.
- the activity of the oligonucleotides is determined in an animal model described in WO 2008/016924, which descriptions are hereby incorporated by reference.
- the oligonucleotide may exhibit at least about 50% target miRNA inhibition or target de-repression at a dose of about 50 mg/kg or less, about 25 mg/kg or less, such as about 10 mg kg or less or about 5 mg/kg or less.
- the oligonucleotide may be dosed intravenously or subcutaneously to mice or to rat, and the oligonucleotide may be formulated in saline.
- the oligonucleotides of the invention are stable after administration, being detectable in the circulation and/or target organ for at least about three weeks, at least about four weeks, at least about five weeks, or at least about six weeks, or more, following administration.
- the oligonucleotides of the invention have the potential to provide for less frequent admin stration, lower doses, and/or longer duration of therapeutic effect.
- the oligonucleotide comprises the nucleotide sequence 5 '-GTGCTGCT-3 ' and is substantially complementary to a nucleotide sequence of human miR-15a, mi.R-15b, miR- 16, miR-195, miR-424, and miR-497.
- the oligonucleotide further contains at least one locked nucleotide(s).
- the oligonucleotide may contain at least eight or at least nine locked nucleotides.
- the length of the oligonucleotide and number and position of locked nucleotides is such that the oligonucleotide reduces the activity of miR- 15a, miR-15b, miR-16, miR-195, miR-424, and miR-497 at an oligonucleotide concentration of about 50 nM or less in the in vitro luciferase assay, or at a dose of about 50 mg/kg or less, or about 25 mg/kg or less in a suitable mouse or rat model, each as described herein.
- a substantially complementary oiigonucleotide may have from about 1 to about 5 mismatches (e.g., 1 or about 2 or about 3 or about 4 or about 5 mismatches) with respect to its target sequence of miR-15a, miR-15b, miR-16, miR-195, miR-424, and miR-497.
- miR-15 family members including miR-15a, miR- 15b, miR-16, miR-195, miR-424, and miR-497, as well as their potential for treating cardiac hypertrophy, heart failure, or myocardial infarction (among other conditions), are described in WO 2009/062169, which is hereby incorporated by reference.
- the pre- miR A sequences for human miR- 15 family members e.g. stem loop sequences
- which may be used for designing inhibitory miRNAs in accordance with the invention are listed below: Human pre-miR- 15a CCUUGGAGUA AAGUAGCAGC ACAUAAUGGU UUGUGGAUUU UGAAAAGGUG
- CAAGAUGCGA AUCAUUAUUU GCUGCUCUAG AAAUUUAAGG AAAUUCAU (SEQ ID 14)
- Each of the pre-miRNA sequences for each miR-15 family member is processed into a mature sequence and a star sequence.
- the star sequence is processed from the other strand of the stem loop stnicture.
- the mature and star sequences for each of the miR-15 family members are given below: Human mature miR ⁇ 15a
- the seed region e.g. bases spanning 2 to 8 nucleotides of mature miRNA sequence
- the 3 ' end of the mature miRNA differs among the different family members.
- multiple members of the miR- 15 family may be inhibited simultaneously by administering a single miR-15 inhibitor or by administering multiple inhibitors.
- each inhibitor may target a single miR-15 family member or may target multiple miR-15 family members.
- a single miR-15 inhibitor inhibits the expression or activity of two or more miR-15 family members.
- Such inhibitors include, but are not limited to, M- 10670, M- 1 121 1 , M-1 1213, M- 1 1214, M- 1 1215, V!- l 1 . ?..?. ⁇ ) . M-1 1221 , and M-10222.
- a single miR-15 inhibitor inhibits the expression or activity of three or more miR-15 family members.
- Such inhibitors include, but are not limited to, M- 10670, M-1 121 1 , M-1 1213, M-1 1214, M-1 1215, M-1 1220, M-1 1221 , and M- 10222.
- a single miR-15 inhibitor inhibits the expression or activity of four or more miR-15 family members.
- Such inhibitors may include, but are not limited to, M-1 121 1 , M-1 1213, M-1 1214, M-1 1215, M-1 1220, and M-1 1221.
- the oligonucleotide contains one or more locked nucleic acid (LNAs) residues, or
- LNAs locked nucleotides. LNAs are described, for example, in US Patent 6,268,490, US Patent No. 6,316, 198, US Patent No. 6,403,566, US Patent No. 6,770,748, US Patent No. 6,998,484, US Patent No. 6,670,461 , and US Patent No. 7,034, 133, ail of which are hereby incorporated by reference in their entireties. LNAs are modified nucleotides or ribonucleotides that contain an extra bridge between the 2' and 4' carbons of the ribose sugar moiety resulting in a "locked" conformation, and/or bicyclic structure.
- the oligonucleotide contains one or more LNAs having the structure shown by structure A below.
- the oligonucleotide may contain one or more LNAs having the structure shown by structure B below.
- the oligonucleotide contains one or more LNAs having the structure shown by structure C below.
- Suitable locked nucleotides that can be incorporated in the oligonucleotides of the invention include those described in US Patent No. 6,403,566 and US Patent No. 6,833,36! , both of which are hereby incorporated by reference in their entireties.
- the locked nucleotides have a 2' to 4' methylene bridge, as shown in structure A, for example.
- the locked nucleotides have a bridge comprising an ethylene group, which may or may not contain an ether linkage at the 2" position.
- the oligonucleotide may comprise, consist essentially of, or consist of, a full length or truncated antisense sequence of a miR-15 family member.
- the term "full length" in reference to a miRNA sequence refers to the length of the mature miRNA antisense counterpart.
- the inhibitors described herein may be truncated or full-length, antisense, mature miRNA sequences, or may comprise these sequences in combination with other polynucleotide sequences.
- the chemical modification motif described herein renders full length antisense miRNA (mature) sequences unnecessary.
- the oligonucleotide is from about 8 to about 20 nucleotides in length, or is from about 8 to about 18 nucleotides in length, or is from about 12 to about 17 nucleotides in length.
- the oligonucleotide in some embodiments is about 8, about 9, about 10, about 1 1 , about 12, about 13, about 14, about 15, about 16 nucleotides in length.
- the truncated oligonucleotide may have a sequence that targets, by antisense inhibition, a miR-15a sequence within 5'- UAGCAGCACAUAAUGGU-3 ' (SEQ ID NO.
- a miR-15b sequence within 5 '- UAGCAGCACAUCAUGGU-3 ' (SEQ ID NO. 34), a miR- 16 sequence within 5 '- UAGCAGCACGUAAAUAU-3 ' (SEQ ID NO. 35), a miR-195 sequence within UAGC AGCAC AGAAAUAU-3 ' (SEQ ID NO. 36), a miR-424 sequence within 5'- CAGCAGCAAUUCAUGUU-3 ' (SEQ ID NO. 37), or a miR-497 sequence within 5'- CAGCAGCACACUGUGGU-3 ' (SEQ ID NO. 38).
- the oligonucleotide generally has a nucleotide sequence designed to target mature miR-I5a, miR-15b, miR-16, miR-195, miR-424, and miR-497.
- the oligonucleotide may, in these or other embodiments, also or alternatively be designed to target the pre- or pri- miRNA forms.
- the oligonucleotide may be designed to have a sequence containing from about 1 to about 5 (e.g., about 1 , about 2, about 3, or about 4) mismatches relative to the fully complementary (mature) miR- 15 sequence.
- such antisense sequences may be incorporated into shRNAs or other R A structures containing stem and loop portions, for example.
- the oligonucleotide is about 16 nucleotides in length.
- the oligonucleotide may comprise or consist essentially of a nucleotide sequence selected from 5 ' - ACCATT ATGTGCTGCT-3 ' (SEQ ID NO. 1), 5 ' - ACC ATGATGTGCTGCT-3 ' (SEQ ID NO. 2), 5 '-ATATTTACGTGCTGCT-3 ' (SEQ ID NO. 3), and 5 '- ATATTTCTGTGCTGCT-3 ' (SEQ ID NO. 4).
- the oligonucleotide may include a mix of locked and non-locked nucleotides.
- the oligonucleotide may include nine locked nucleotides and seven non-locked nucleotides.
- the pattern of locked nucleotides may be such that at least positions 1 , 5, 8, 10, and 16 are locked nucleotides.
- the oligonucleotide is about 12 nucleotides in length.
- the oligonucleotide may comprise or consist essentially of a nucleotide sequence selected from 5'- TT ATGTGCTGCT-3 ' (SEQ ID NO. 5), 5 ' -TT ACGTGCTGCT-3 ' (SEQ ID NO. 6), 5 ' -TTCTGTGCTGCT-3 ' (SEQ ID NO.
- the oligonucleotide may include a mix of locked and non-locked nucleotides.
- the oligonucleotide may include eight locked nucleotides and four non-locked nucleotides.
- the pattern of locked nucleotides may be such that at least positions 1, 4, 9, and 12 are locked nucleotides.
- the oligonucleotide is about 8 nucleotides in length.
- the oligonucleotide may consist essentially of, or consists of the nucleotide sequence 5'- GTGCTGCT-3', where all or substantially all (e.g., at least about 6 or at least about 7) are locked nucleotides.
- the oligonucleotide generally contains at least about 8 locked nucleotides, but in various embodiments is not fully comprised of locked nucleotides.
- the number and position of locked nucleotides is such that the oligonucleotide reduces the activity of miR ⁇ 15a, miR-15b, miR-16, miR-195, miR-424, and miR-497 as determined in vitro or in vivo as described.
- the oligonucleotide does not contain a stretch of nucleotides with more than about four, or more than about three, contiguous non-locked nucleotides, in these or other embodiments, the region complementary to the miR-15a, miR-15b, miR-16, miR-195, miR-424, and miR-497 seed region comprises at least three or at least four locked nucleotides.
- the nucleotide may contain a 2 " modification with respect to a 2' hydroxy!.
- the 2' modification may be 2' deoxy.
- Incorporation of 2 -modified nucleotides in antisense oligonucleotides may increase both resistance of the oligonucleotides to nucleases and their thermal stability with complementary RNA.
- Various modifications at the 2' positions may be independently selected from those that provide increased nuclease sensitivity, without compromising molecular interactions with the RNA target or cellular machinery. Such modifications may be selected on the basis of their increased potency in vitro or in vivo. Exemplary methods for determining increased potency (e.g., IC50) for miRNA inhibition are described herein, including the dual luciferase assay and in vivo miRNA expression or target de-repression.
- the 2' modification may be independently selected from O- alkyl (which may be substituted), halo, and deoxy (H).
- nucleotide 2' positions of the non-locked nucleotides may be modified in certain embodiments, e.g., as independently selected from O-aikyi (e.g., O-methyl), halo (e.g., fluoro), deoxy (H), and amino.
- the 2 " modifications may each be independently selected from O-methyl and fluoro.
- purine nucleotides each have a 2' OMe and pyrimidine nucleotides each have a 2'-F. In certain embodiments, from one to about five 2' positions, or from about one to about three 2' positions are left unmodified (e.g., as hydroxyls).
- the hydrocarbon substituents include alkyl, alkenyl, alkynyl, and alkoxyalkyl, where the alkyl (including the alkyl portion of aikoxy), alkenyl and alkynyl may be substituted or unsubstituted.
- the alkyl, alkenyl, and alkynyl may be CI to CIO alkyl, alkenyl or alkynyl, such as CI, C2, or C3.
- the hydrocarbon substituents may include one or two or three non-carbon atoms, which may be independently selected from N, O, and/or S.
- the 2' modifications may further include the alkyl, alkenyl, and alkynyl as O-alkyl, O-alkenyl, and O-alkynyl.
- Exemplary modifications in accordance with the invention include 2' -O-alkyl (CI -3 alkyl, such as 2'OMe or 2'OEt), 2'-0-methoxyethyl (2'-0 ⁇ MOE), 2'-0-ammopropyl (2'-0-AP), 2'-0-dimemylaminoethyl (2'-0-DMAOE), 2'-0-dime laminopropyl (2'-0- DMAP), 2'-0-dimethyiaminoethyloxyethyi (2 -O-DMAEOE), or 2'-0-N-methyiacetamido (2'-0-NMA) substitutions.
- the oligonucleotide contains at least one 2 '-halo modification (e.g., in place of a hydroxy!), such as 2' -fluoro, 2'-chloro, 2'-bromo, and 2'-iodo.
- the 2' halo modification is fluoro.
- the oligonucleotide may contain from 1 to about 5 2' -halo modifications (e.g., fluoro), or from 1 to about 3 2'- halo modifications (e.g., fluoro).
- the oligonucleotide contains all 2 ' - Hiioro nucleotides at non-locked positions, or 2'-fluoro on all non-locked pvrimidine nucleotides.
- the 2' ⁇ fluoro groups are independently di-, tri-, or un-methylated.
- the oligonucleotide may have one or more 2'-deoxy modifications (e.g., H for 2' hydroxy 1), and in some embodiments, contains from about 2 to about 10 2'-deoxy modifications at non-iocked positions, or contains 2'deoxy at ail non-locked positions.
- 2'-deoxy modifications e.g., H for 2' hydroxy 1
- the oligonucleotide contains 2' positions modified as 2'OMe in non-locked positions.
- non-iocked purine nucleotides are modified at the 2' position as 2'OMe, with non-iocked pvrimidine nucleotides modified at the 2" position as 2'-fluoro.
- the oligonucleotide further comprises at least one terminal modification or "cap".
- the cap may be a 5' and/or a 3 -cap structure.
- the terms “cap” or “end-cap” include chemical modifications at either terminus of the oligonucleotide (with respect to terminal ribonucleotides), and including modifications at the linkage between the last two nucleotides on the 5' end and the last two nucleotides on the 3' end.
- the cap structure as described herein may increase resistance of the oligonucleotide to exonucleases without compromising molecular interactions with the RNA. target or cellular machinery. Such modifications may be selected on the basis of their increased potency in vitro or in vivo.
- the cap can be present at the 5'-terminus (5'-cap) or at the 3 - terminus (3 -cap) or can be present on both ends.
- the 5'- and/or 3 '-cap is independently selected from phosphorothioate monophosphate, abasic residue (moiety), phosphorothioate linkage, 4'-thio nucleotide, carbocyclic nucleotide, phosphorodithioate linkage, inverted nucleotide or inverted abasic moiety (2'-3' or 3 '-3'), phosphorodithioate monophosphate, and methylphosphonate moiety.
- the phosphorothioate or phosphorodithioate linkage(s), when part of a cap structure, are generally positioned between the two terminal nucleotides on the 5' end and the two terminal nucleotides on the 3 ' end.
- the oligonucleotide has at least one terminal phosphorothioate monophosphate.
- the phosphorothioate monophosphate may support a higher potency by inhibiting the action of exonucleases.
- the phosphorothioate monophosphate may be at the 5' and/or 3' end of the oligonucleotide.
- a phosphorothioate monophosphate is defined by the following structures, where B is base, and R is a 2' modification as described above:
- the cap structure may incorporate a locked nucleotide as described herein,
- Phosphorothioate linkages may be present in some embodiments, such as between the last two nucleotides on the 5' and the 3' end (e.g., as part of a cap structure), or as alternating with phosphodiester bonds.
- the oligonucleotide may contain at least one terminal abasic residue at either or both the 5' and 3' ends.
- An abasic moiety does not contain a commonly recognized purine or pyrimidine nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine.
- abasic moieties lack a nucleotide base or have other non-nucleotide base chemical groups at the 1 ' position.
- the abasic nucleotide may be a reverse abasic nucleotide, e.g., where a reverse abasic phosphoramidite is coupled via a 5' amidite (instead of 3' amidite) resulting in a 5 '-5' phosphate bond .
- the structure of a reverse abasic nucleoside for the 5' and the 3' end of a polynucl eotide is shown below.
- the oligonucleotide may contain one or more phosphorothioate linkages. Phosphorothioate linkages have been used to render oligonucleotides more resistant to nuclease cleavage.
- the polynucleotide may be partially phosphorothioate- linked, for example, phosphorothioate linkages may alternate with phophodiester linkages. In certain embodiments, however, the oligonucleotide is fully phosphorothioate-linked , In other embodiments, the oligonucleotide has from about one to about five or about one to about three phosphate linkages.
- the nucleotide has one or more earboxamido-modified bases as described in WO 2012/061810, which is hereby incorporated by reference, including with respect to all exemplary pyrrolidine carboxamido modifications disclosed therein with heterocyclic substituents.
- the oligonucleotide has the structure of a compound listed in Table 1 , below. As depicted in Table 1 , a plus sign (e.g., +A) indicates a LNA base. All other bases are DNA. Each miR-I5 inhibitor has a folly phosphorothioate linked backbone. Table 1 : Exemplaiy Oligonucleotides
- oligonucleotides including modified polynucleotides, by solid phase synthesis is well known and is reviewed in New Chemical Methods for Synthesizing Polynucleotides.
- the oligonucleotide may be incorporated within a variety of macromolecular assemblies or compositions. Such complexes for delivery may include a variety of liposomes, nanoparticles, and micelles, formulated for delivery to a patient.
- the complexes may include one or more fusogenic or lipophilic molecules to initiate cellular membrane penetration. Such molecules are described, for example, in US Patent No. 7,404,969 and US Patent No. 7,202,227, which are hereby incorporated by reference in their entireties.
- the oligonucleotide may further comprise a pendant lipophilic group to aid cellular delivery, such as those described in WO 2010/129672, which is hereby incorporated by reference.
- composition or formulation may employ a plurality of therapeutic oligonucleotides, including at least one described herein.
- the composition or formulation may employ at least about 2, about 3, about 4, or about 5 miRNA inhibitors described herein.
- the oligonucleotides of the invention may be formulated as a variety of pharmaceutical compositions.
- Pharmaceutical compositions will be prepared in a form appropriate for the intended application. Generally, this will entail preparing compositions that are essentially free of pyrogens, as well as other impurities that could be harmful to humans or animals.
- Exemplary delivery/formulation systems include colloidal dispersion systems, macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in- water emulsions, micelles, mixed micelles, and liposomes.
- fat emulsions that are suitable for delivering the nucleic acids of the invention to cardiac and skeletal muscle tissues include Intralipid®, Liposyn®, Liposyn® II, Liposyn® III, Nutrilipid, and other similar lipid emulsions.
- a preferred colloidal system for use as a deliver ⁇ ' vehicle in vivo is a liposome (i.e., an artificial membrane vesicle). The preparation and use of such systems is well known in the art. Exemplary formulations are also disclosed in US Patent No. 5,981 ,505; US Patent No. 6,217,900; US Patent No. 6,383,512; US Patent No. 5,783,565; US Patent No. 7,202,227; US Patent No. 6,379,965; US Patent No. 6,127,170; US Patent No. 5,837,533; US Patent
- the oligonucleotide is formulated for conventional subcutaneous or intravenous administration, for example, by formulating with appropriate aqueous diluent, including sterile water and normal saline.
- compositions and formulations may employ appropriate salts and buffers to render delivery vehicles stable and allow for uptake by target cells.
- Aqueous compositions of the present invention comprise an effective amount of the delivery vehicle comprising the inhibitor oligonucleotide (e.g. liposomes or other complexes), dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.
- pharmaceutically acceptable or “pharmacologically acceptable” refers to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal or a human.
- pharmaceutically acceptable carrier may include one or more solvents, buffers, solutions, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like acceptable for use in formulating pharmaceuticals, such as pharmaceuticals suitable for administration to humans.
- pharmaceuticals such as pharmaceuticals suitable for administration to humans.
- the use of such media and agents for pharmaceutically active substances is well known in the art.
- Supplementary active ingredients also can be incorporated into the compositions.
- Administration or delivery of the pharmaceutical compositions according to the present invention may be via any route so long as the target tissue is available via that route.
- admin stration may be by intradermal, subcutaneous, intramuscular, intraperitoneal or intravenous injection, or by direct injection into target tissue (e.g., cardiac tissue).
- compositions comprising miRNA inhibitors may also be administered by catheter systems or systems that isolate coronary circulation for delivering therapeutic agents to the heart.
- catheter systems for delivering therapeutic agents to the heart and coronary vasculature are known in the art.
- Some non-limiting examples of catheter-based delivery methods or coronary isolation methods suitable for use in the present invention are disclosed in U.S. Patent No. 6,416,510; U.S. Patent No. 6,716,196; U.S. Patent No. 6,953,466, WO 2005/082440, WO 2006/089340, U.S. Patent Publication No.
- compositions or formulations may also be administered parenteral! ⁇ ' or intraperitoneal [y.
- solutions of the conjugates as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose.
- Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations generally contain a preservative to prevent the growth of microorganisms.
- the pharmaceutical forms suitable for injectable use or catheter deliveiy include, for example, sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
- these preparations are sterile and fluid to the extent that easy injectabiiity exists.
- Preparations should be stable under the conditions of manufacture and storage and should be preserved against the contaminating action of microorganisms, such as bacteria and fungi.
- Appropriate solvents or dispersion media may contain, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof!, and vegetable oils.
- the proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
- a coating such as lecithin
- surfactants for example, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium sorbic acid, thimerosai, and the like.
- isotonic agents for example, sugars or sodium chloride.
- Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
- Sterile injectable solutions may be prepared by incorporating the conjugates in an appropriate amount into a solvent along with any other ingredients (for example as enumerated above) as desired.
- dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the desired other ingredients, e.g., as enumerated above.
- the preferred methods of preparation include vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient(s) plus any additional desired ingredient from a previously sterile-filtered solution thereof.
- solutions are preferably administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective.
- the formulations may easily be administered in a variety of dosage forms such as injectable solutions, drug release capsules and the like.
- aqueous solution for example, the solution generally is suitably buffered and the liquid diluent first rendered isotonic for example with sufficient saline or glucose.
- aqueous solutions may be used, for example, for intravenous, intramuscular, subcutaneous and intraperitoneal administration.
- sterile aqueous media are employed as is known to those of skill in the art, particular!)' in light of the present disclosure.
- a single dose may be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoc lysis fluid or injected at the proposed site of infusion, (see for example, "Remington's Pharmaceutical Sciences” 15th Edition, pages 1035-1038 and 1570-1580).
- Some variation in dosage will necessarily occur depending on the condition of the subject being treated.
- the person responsible for administration will, in any event, determine the appropriate dose for the individual subject.
- preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologies standards.
- the invention provides a method for delivering oligonucleotides to a mammalian cell (e.g., as part of a composition or formulation described herein), and methods for treating, ameliorating, or preventing the progression of a condition in a mammalian patient.
- the oligonucleotide or pharmaceutical composition may be contacted in vitro or in vivo with a target cell (e.g., a mammalian cell).
- the cell may be a heart cell.
- the method generally comprises administering the oligonucleotide or composition comprising the same to a mammalian patient or population of target cells.
- the oligonucleotide as already described, is a iR A inhibitor (e.g., having a nucleotide sequence designed to inhibit expression or activity of a miR-15 family miRNA).
- the patient may have a condition associated with, mediated by, or resulting from, miR-15 family expression.
- miR-15 family miRNA e.g., having a nucleotide sequence designed to inhibit expression or activity of a miR-15 family miRNA.
- the invention provides a use of the modified oligonucleotides and compositions of the invention for treating such conditions, and for the preparation of medicaments for such treatments.
- the patient e.g.
- human patient has one or more risk factors including, for example, long standing uncontrolled hypertension, uncorrected valvular disease, chronic angina, recent myocardial infarction, congestive heart, failure, congenital predisposition to heart disease and pathological hypertrophy.
- risk factors including, for example, long standing uncontrolled hypertension, uncorrected valvular disease, chronic angina, recent myocardial infarction, congestive heart, failure, congenital predisposition to heart disease and pathological hypertrophy.
- the patient may have been diagnosed as having a genetic predisposition to, for example, cardiac hypertrophy, or may have a familial history of, for example, cardiac hypertrophy.
- administering results in the improvement of one or more symptoms of, for example, cardiac hypertrophy, heart failure, ischemia, ischemia reperfusion injury, or myocardial infarction in the subject, or in the delay in the transition from cardiac hypertrophy to heart failure.
- the one or more improved symptoms may be, for example, increased exercise capacity, increased cardiac ejection volume, decreased left ventricular end diastolic pressure, decreased pulmonary capil lary wedge pressure, increased cardiac output, increased cardiac index, lowered pulmonary artery pressures, decreased left ventricular end systolic and diastolic dimensions, decreased left and right ventricular wail stress, decreased wall tension, increased quality of life, and decreased disease related morbidity or mortality.
- use of inhibitors of miR-15 family members may prevent cardiac hypertrophy and its associated symptoms from arising.
- an mhibitor of the miR- 15 family members may be administered in combination with other therapeutic modalities.
- a miR-15 inhibitor of the invention may be administered in conjunction with other types of therapeutic agents such as anti-hyperlipoproteinemic agents, anti-arteriosclerotic agents, an ti-thrombotic/fibrino lytic agents, blood coagulants, anti-arrhythmic agents, antihypertensive agents, treatment agents for congestive heart failure, anti-anginal agents, anti-bacterial agents, vasodilators, hormone antagonists, iontropes, diuretics, endothelin receptor antagonists, calcium channel blockers, phosphodiesterase inhibitors, angiotensin II converting enzyme (ACE) inhibitors, cytokine blockers/ nhibitors, HDAC inhibitors or a combination thereof.
- ACE angiotensin II converting enzyme
- the combination therapy also may involve inhibiting the expression or activity of additional miRNAs involved in cardiac remodeling such as miR- 499, miR-208, miR-208b and miR-21, which are described in WO 2012/083005 and WO2009/058818, the contents of which are hereby incorporated by reference.
- a miR- 15 inhibitor may be administered in conjunction with surgery.
- a miR-! 5 inhibitor may also be administered together with non-pharmacological means of treatment.
- non-pharmacological treatment may involve reducing sodium in the diet,
- the pharmaceutical composition is administered by parenteral administration or by direct injection into heart tissue.
- the parenteral administration may be intravenous, subcutaneous, or intramuscular.
- the composition is administered by oral, transdermal, sustained release, controlled release, delayed release, suppository, catheter, or sublingual administration.
- the oligonucleotide is administered at a dose of about 25 mg/kg or less, or a dose of about 10 mg/kg or less, or a dose of about 5 mg/kg or less.
- the oligonucleotide or composition may be administered by intramuscular or subcutaneous injection, or intravenously.
- the activity of miR- 15a, miR- 15b, miR- 16, miR- 195, miR-424, and miR-497 in cardiac tissue, or as determined in patient serum is reduced or inhibited.
- the method of the invention may comprise administering a miR- 15 family inhibitor, which may be an oligonucleotide described herein, to a population of target cells or a mammalian subject at a dose effective for target de- repression of Piml .
- the present invention also provides method of regulating expression of Piml and other target genes such as Bcl2L2, Birc5, Grn, and Cdc2A in a cell comprising contacting the cell with a miR- 15 inhibitor.
- the expression of Pim! , Bcl2L2, Birc5, Grn, and Cdc2A are increased following administration of a miR- 15 inhibitor.
- Expression of Piml and/or other markers described herein may be monitored before, during, or after treatment to determine treatment response.
- the methods further comprise scavenging or clearing the miRNA inhibitors following treatment.
- an oligonucleotide having a nucleotide sequence that is complementary to the inhibitor may be administered after therapy to attenuate or stop the function of the inhibitor.
- Example 1 Abundance of miR- 15 family members in cardiac tissue
- FIG. 1 The results ( Figure 1) show that miR- 16 is the most abundant of the mi.R-15 family members in cardiac tissue, with approximately 10,000 copes per cell. Other miR- 15 family members including miR-15a, miR-15b, miR-195, miR-497, miR-424, and miR- 322 are expressed at approximately 1,000 copies per cell. In addition, miR-322 appears to be only expressed in rodents while miR-424 expression appears to be restricted to larger animals (e.g., human and pigs).
- Example 2 Identification of gene targets regulated by miR- 15 family members
- a panel of miRNA inhibitors single stranded oligonucleotides
- the sequences and modification patterns are shown in Table 1.
- the panel included multiple lengths of reverse complement inhibitors ranging from 8 nucleotides to 16 nucleotides.
- the number of LNA modifications was varied as well as the location of the LNA modifications in the oligonucleotide.
- wild-type C57/BL6 mice were subcutaneously injected with either saline, or 2.8 mg/kg, 10 mg/kg, or 25 mg kg of a control oligonucleotide or an anti-miRl 5b oligonucleotide (M-10134). Three doses were given during a treatment period of three days. Cardiac tissue was harvested at 24 hours following the last injection, and target gene expression was assessed by real-time PCR.
- Example 3 Activit of mi RN A inhibitors targeting the mill- 15 fami ly Twenty five miRNA inhibitors (as shown in Table 1 ) were tested for their ability to inhibit miR-15 family members.
- wild-type C57/BL6 mice were subcutaneously injected with either saline, or 25 mg/kg of a control oligonucleotide or individual anti-miR15 oligonucleotides. Three doses were given during a treatment period of three days. Cardiac tissue was harvested at 24 hours following the last injection, and target gene expression was assessed by real-time PCR. As shown in Figure 4, the length of the rniRNA inhibitor as well, as LNA patterns impart distinct inhibitory activities.
- inhibitors M-10670, M-1121 1 , M-1 1213, M- 11214, M-11215, M-11220, M-11221, M-11222 strongly affect the expression of miR- 15 target genes such as Birc5, Cdc2A, Grn, CcnDl, and CcnD2. These inhibitors vary from 12 nucleotides in length to 16 nucleotides in length. Of these, the 12-mer inhibitor M- 1 1211 appears to exhibit the most potent inhibitory activity, affecting the expression of four out of five miR- 15 target genes.
- a two-step real-time PCR assay (Applied Biosystems) was utilized to assess the ability of rniRNA. inhibitors to inhibit individual miR- 15 family members. Specifically, the potent compounds identified previously including M-10670, M-1 1211, M-11213, M- 11214, M-11215, M-11220, M-11221, M-11222 were tested for their effects on miR- 15a, miR- 15b, miRl 6, and mi.R-195 expression. Wild-type C57/BL6 mice were
- rniRNA inhibitors Activities of the rniRNA inhibitors were tested in vitro utilizing a dual-l ciferase assay. Specifically, HeLa cells were transfected with varying concentrations of a miR- 15 inhibitor as well as 25 ng/well of a reporter plasmid. In particular, the miR- 195 dual luciferase construct did not appear to be as sensitive as the other constmcts with regard to microRNA inhibition, as demonstrated by the need to transfect inhibitor concentrations in the range of 2 to 50 nM, compared to 0.31 to 8.3 iiM for the other constmcts.
- Example 4 In vivo activity of miRNA inhibitors targeting the miR-15 family
- mice were subcutaneously injected with 25 mg/kg of M-11215. Only a single dose was administered. Cardiac tissue was harvested at either 24 hours, 48 hours, 72 hours, or 98 hours post injection. As shown in Figure 10, a single dose of M-11215 elicits de-repression of target genes such as Birc5 as early as 24 hours post treatment.
- C Global and specific target de-repression by miRNA. inhibitors
- Example 5 In vivo activity of miR-15 inhibitors in rats
- the in vivo activities of the miR-15 inhibitors were further tested in rats. More specifically, Sprague-Dawley rats (49 to 52 days of age) were injected with 25 mg/kg of miR-15 inhibitors including M- 10670, M-l 121 1 , M- 1 1213, M-1 1214, M-l 1215, M- 1 1220, M-l 1221 , or M-l 1222. Only a single dose was administered. Cardiac tissues were harvested at day four post injection, and target gene expression was analyzed. The results show (Figure 12A) that 16-mer inhibitors appear to have a more potent effect on target gene expression in rats.
- CPCs cardiac progenitor cel ls
- CPCs are self-renewing cel ls that produce daughter cells which supply the heart with new myocytes and vessels, thereby stimulating myocardial regeneration.
- the link between miR-15 and Piml suggests that inhibition of miR-15 results in the production of new cardiomyocytes, which can potential ly be beneficial for repairing cardiac damage.
- Example 6 In vivo activity of miR-15 inhibitors in a rat ischemia-reperfusion injury model
- the in vivo activities of the miR-15 inhibitors were tested in a rat ischemia- reperfusion injury model. Specifically, in the ischemia-reperfusion model, the rats were injected intravenously with a single dose (25 mg/kg) of miR-15 inhibitors including M ⁇ 10670, M-l 121 1 , M-1 1214, or M-10591. The rats were sacrificed at 72 hours after reperfusion, and tissues were harvested and analyzed for target gene expression (as measured by real-time PGR). As shown in Figure 13, all miR-15 mhibitors induced significant de-repression of target genes including Birc5, Cdc2a, and Granulin when compared to non-infarcted controls (baseline).
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Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016022536A2 (en) | 2014-08-04 | 2016-02-11 | MiRagen Therapeutics, Inc. | Inhibitors of myh7b and uses thereof |
US9487783B2 (en) | 2014-08-07 | 2016-11-08 | Regulus Therapeutics Inc. | Targeting microRNAs for metabolic disorders |
WO2017219170A1 (en) * | 2016-06-19 | 2017-12-28 | 毛侃琅 | Construction and application of lentiviral vector for specifically inhibiting human mirna-29a and mir-424 expression |
US10337005B2 (en) | 2012-06-21 | 2019-07-02 | MiRagen Therapeutics, Inc. | Oligonucleotide-based inhibitors comprising locked nucleic acid motif |
WO2021020412A1 (en) | 2019-07-30 | 2021-02-04 | 塩野義製薬株式会社 | Nucleic acid drug targeting murf1 |
WO2022018269A1 (en) * | 2020-07-23 | 2022-01-27 | Johann Wolfgang Goethe-Universität Frankfurt | Combinatorial inhibition of mirnas for treatment of heart failure |
EP4035659A1 (en) | 2016-11-29 | 2022-08-03 | PureTech LYT, Inc. | Exosomes for delivery of therapeutic agents |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10011833B2 (en) | 2013-03-15 | 2018-07-03 | MiRagen Therapeutics, Inc. | Bridged bicyclic nucleosides |
US10443044B2 (en) | 2014-04-17 | 2019-10-15 | Ips Heart | Generating cardiac progenitor cells from pluripotent stem cells using isoxazole or isoxazole like compounds |
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US10941403B2 (en) | 2018-04-02 | 2021-03-09 | Oregon Health & Science University | Microrna inhibitors as anti-cancer therapeutics |
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090175827A1 (en) * | 2006-12-29 | 2009-07-09 | Byrom Mike W | miR-16 REGULATED GENES AND PATHWAYS AS TARGETS FOR THERAPEUTIC INTERVENTION |
US20100298410A1 (en) * | 2007-10-04 | 2010-11-25 | Santaris Pharma A/S | MICROMIRs |
US20100317713A1 (en) * | 2007-11-09 | 2010-12-16 | Eric Olson | Micro-rnas of the mir-15 family modulate cardiomyocyte survival and cardiac repair |
Family Cites Families (88)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003029459A2 (en) | 2001-09-28 | 2003-04-10 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Micro-rna molecules |
WO2003093441A2 (en) | 2002-05-03 | 2003-11-13 | Duke University | A method of regulating gene expression |
US7892793B2 (en) | 2002-11-04 | 2011-02-22 | University Of Massachusetts | Allele-specific RNA interference |
AU2003291433B2 (en) | 2002-11-13 | 2008-05-22 | Thomas Jefferson University | Compositions and methods for cancer diagnosis and therapy |
US7482117B2 (en) | 2002-12-20 | 2009-01-27 | Celera Corporation | Genetic polymorphisms associated with myocardial infarction, methods of detection and uses thereof |
CN1768139A (en) | 2003-02-10 | 2006-05-03 | 独立行政法人产业技术总合研究所 | Regulation of gene expression by DNA interference |
WO2005017111A2 (en) | 2003-07-15 | 2005-02-24 | The Trustees Of The University Of Pennsylvania | Methods and systems for identifying micro-rna targets and synthesizing novel micro-rnas and uses of the same |
EP1648914A4 (en) | 2003-07-31 | 2009-12-16 | Regulus Therapeutics Inc | Oligomeric compounds and compositions for use in modulation of small non-coding rnas |
US8106180B2 (en) | 2003-08-07 | 2012-01-31 | Whitehead Institute For Biomedical Research | Methods and products for expression of micro RNAs |
WO2005017145A1 (en) | 2003-08-13 | 2005-02-24 | Japan Biological Informatics Consortium | Method of identifying or presuming gene under regulation regulated by functional rna and method of using the same |
EP1676914A4 (en) | 2003-09-22 | 2007-10-10 | Aichi Prefecture | Disease type of lymphoma and method of assessing prognosis |
JP2007512805A (en) | 2003-10-14 | 2007-05-24 | ノバルティス アクチエンゲゼルシャフト | Oligonucleotide microarray |
US20050182011A1 (en) | 2003-11-13 | 2005-08-18 | Eric Olson | Inhibition of TRP channels as a treatment for cardiac hypertrophy and heart failure |
KR100860087B1 (en) | 2003-12-15 | 2008-09-25 | 포천중문의과대학교 산학협력단 | Novel mirna molecules isolated from human embryonic stem cell |
US20050256072A1 (en) | 2004-02-09 | 2005-11-17 | University Of Massachusetts | Dual functional oligonucleotides for use in repressing mutant gene expression |
EP2295604B1 (en) | 2004-02-09 | 2015-04-08 | Thomas Jefferson University | Diagnosis and treatment of cancers with microRNA located in or near cancer-associated chromosomal features |
AU2005214904B2 (en) | 2004-02-13 | 2011-07-21 | Rockefeller University | Anti-microRNA oligonucleotide molecules |
US8088902B2 (en) | 2004-04-05 | 2012-01-03 | The Rockefeller University | DNA virus microRNA and methods for inhibiting same |
US20060134639A1 (en) | 2004-04-06 | 2006-06-22 | Huffel Christophe V | Method for the determination of cellular transcriptional regulation |
US20050260648A1 (en) | 2004-04-06 | 2005-11-24 | Huffel Christophe V | Method for the determination of cellular transcriptional |
ES2380923T3 (en) | 2004-04-07 | 2012-05-21 | Exiqon A/S | Small interfering microRNA and RNA quantification procedures |
EP2290071B1 (en) | 2004-05-28 | 2014-12-31 | Asuragen, Inc. | Methods and compositions involving microRNA |
CA2576233C (en) | 2004-08-10 | 2016-03-15 | Alnylam Pharmaceuticals, Inc. | Conjugate comprising an antagomir and a ligand |
DK1797183T3 (en) | 2004-09-02 | 2012-10-01 | Univ Yale | REGULATION OF ONCOGENES WITH MICRORNAS |
FR2877350B1 (en) | 2004-11-03 | 2010-08-27 | Centre Nat Rech Scient | IDENTIFICATION AND USE OF miRNAs INVOLVED IN THE DIFFERENTIATION OF CELLS FROM MYELOID LEUKEMIA |
WO2006137941A2 (en) | 2004-11-12 | 2006-12-28 | Ambion, Inc. | Methods and compositions involving mirna and mirna inhibitor molecules |
WO2006063356A1 (en) | 2004-12-10 | 2006-06-15 | Isis Phamaceuticals, Inc. | Regulation of epigenetic control of gene expression |
US20060185027A1 (en) | 2004-12-23 | 2006-08-17 | David Bartel | Systems and methods for identifying miRNA targets and for altering miRNA and target expression |
US20060166920A1 (en) | 2005-12-27 | 2006-07-27 | Regents Of The University Of Michigan | Oligonucleotide based therapeutics |
EP1838870A2 (en) | 2004-12-29 | 2007-10-03 | Exiqon A/S | NOVEL OLIGONUCLEOTIDE COMPOSITIONS AND PROBE SEQUENCES USEFUL FOR DETECTION AND ANALYSIS OF MICRORNAS AND THEIR TARGET MRNAs |
US8071306B2 (en) | 2005-01-25 | 2011-12-06 | Merck Sharp & Dohme Corp. | Methods for quantitating small RNA molecules |
US20070065840A1 (en) | 2005-03-23 | 2007-03-22 | Irena Naguibneva | Novel oligonucleotide compositions and probe sequences useful for detection and analysis of microRNAS and their target mRNAS |
JP2006292367A (en) | 2005-04-05 | 2006-10-26 | Mitsubishi Rayon Co Ltd | Microarray for detecting mirna |
WO2006108582A2 (en) | 2005-04-15 | 2006-10-19 | Cenix Bioscience Gmbh | Human marker genes and agents for diagnosis, treatment and prophylaxis of cardiovascular disorders and artherosclerosis |
US9085774B2 (en) | 2005-04-19 | 2015-07-21 | Basf Plant Science Gmbh | Methods controlling gene expression |
PL2002003T3 (en) | 2005-05-27 | 2016-06-30 | Ospedale San Raffaele Srl | Gene vector comprising mi-rna |
CA2610265A1 (en) | 2005-05-31 | 2007-05-10 | Cold Spring Harbor Laboratory | Methods for producing micrornas |
US20070054287A1 (en) | 2005-05-31 | 2007-03-08 | Applera Corporation | Method for identifying medically important cell populations using micro rna as tissue specific biomarkers |
US20090209621A1 (en) | 2005-06-03 | 2009-08-20 | The Johns Hopkins University | Compositions and methods for decreasing microrna expression for the treatment of neoplasia |
EP1919512B1 (en) | 2005-08-10 | 2014-11-12 | Alnylam Pharmaceuticals Inc. | Chemically modified oligonucleotides for use in modulating micro rna and uses thereof |
US20070213292A1 (en) | 2005-08-10 | 2007-09-13 | The Rockefeller University | Chemically modified oligonucleotides for use in modulating micro RNA and uses thereof |
CN101426912A (en) | 2005-08-17 | 2009-05-06 | 瑟纳治疗公司 | Chemically modified short interfering nucleic acid molecules that mediate RNA interference |
JP2009507918A (en) | 2005-09-12 | 2009-02-26 | ジ・オハイオ・ステイト・ユニバーシティ・リサーチ・ファウンデイション | Compositions and methods for diagnosis and therapy of BCL2-related cancers |
US20070092882A1 (en) | 2005-10-21 | 2007-04-26 | Hui Wang | Analysis of microRNA |
JP2009516518A (en) | 2005-11-21 | 2009-04-23 | ジヨンソン・アンド・ジヨンソン・リサーチ・ピーテイワイ・リミテツド | Multi-targeting interfering RNAs with two active strands and methods for their design and use |
US20070259827A1 (en) | 2006-01-25 | 2007-11-08 | University Of Massachusetts | Compositions and methods for enhancing discriminatory RNA interference |
EP1986609A2 (en) | 2006-01-26 | 2008-11-05 | University of Massachusetts | Rna silencing agents for use in therapy and nanotransporters for efficient delivery of same |
WO2007092181A2 (en) | 2006-01-26 | 2007-08-16 | Unversity Of Massachusetts | Compositions and methods for modulating translational repression |
EP2388328A1 (en) | 2006-01-27 | 2011-11-23 | Isis Pharmaceuticals, Inc. | Oligomeric compounds and compositions for the use in modulation of micrornas |
WO2007095387A2 (en) | 2006-02-17 | 2007-08-23 | Dharmacon, Inc. | Compositions and methods for inhibiting gene silencing by rna interference |
EP2522747A1 (en) | 2006-03-02 | 2012-11-14 | The Ohio State University | MicroRNA expression profile associated with pancreatic cancer |
CA2649045C (en) * | 2006-04-03 | 2019-06-11 | Santaris Pharma A/S | Pharmaceutical composition comprising anti-mirna antisense oligonucleotides |
US20080220423A1 (en) | 2006-05-19 | 2008-09-11 | Soren Moller | Oligonucleotide probes useful for detection and analysis of microRNA precursors |
WO2007147067A2 (en) | 2006-06-14 | 2007-12-21 | Rosetta Inpharmatics Llc | Methods and compositions for regulating cell cycle progression |
US20090307788A1 (en) | 2006-06-19 | 2009-12-10 | Kobenhavns Universitet | Ribozyme mediated stabilization of polynucleotides |
WO2008024499A2 (en) | 2006-08-25 | 2008-02-28 | Duke University | Methods for in vivo identification of endogenous mrna targets of micrornas |
CA2663962A1 (en) | 2006-09-19 | 2008-03-27 | Asuragen, Inc. | Mir-15, mir-26, mir-31,mir-145, mir-147, mir-188, mir-215, mir-216, mir-331, mmu-mir-292-3p regulated genes and pathways as targets for therapeutic intervention |
WO2008042231A2 (en) | 2006-09-29 | 2008-04-10 | Children's Medical Center Corporation | Compositions and methods for evaluating and treating heart failure |
EP2489742A1 (en) | 2006-10-09 | 2012-08-22 | Julius-Maximilians-Universität Würzburg | MicroRNA (miRNA) for the diagnosis and treatment of heart diseases |
WO2008147430A2 (en) | 2006-10-11 | 2008-12-04 | Nucleonics, Inc. | Microrna-formatted multitarget interfering rna vector constructs and methods of using the same |
US20100021914A1 (en) | 2006-11-23 | 2010-01-28 | Querdenker Aps | Oligonucleotides for modulating target rna activity |
CA2671302A1 (en) | 2006-12-08 | 2008-06-19 | Asuragen, Inc. | Mirna regulated genes and pathways as targets for therapeutic intervention |
WO2008073921A2 (en) | 2006-12-08 | 2008-06-19 | Asuragen, Inc. | Mir-126 regulated genes and pathways as targets for therapeutic intervention |
CA2671294A1 (en) | 2006-12-08 | 2008-06-19 | Asuragen, Inc. | Mir-21 regulated genes and pathways as targets for therapeutic intervention |
AU2007333109A1 (en) | 2006-12-08 | 2008-06-19 | Asuragen, Inc. | Functions and targets of let-7 micro RNAs |
US20090137504A1 (en) | 2006-12-21 | 2009-05-28 | Soren Morgenthaler Echwald | Microrna target site blocking oligos and uses thereof |
EP3536788A1 (en) | 2006-12-21 | 2019-09-11 | QIAGEN GmbH | Microrna target site blocking oligos and uses thereof |
US20080287383A1 (en) | 2007-03-02 | 2008-11-20 | Nastech Pharmaceutical Company Inc. | Nucleic acid compounds for inhibiting erbb gene expression and uses thereof |
US9043994B2 (en) | 2007-03-13 | 2015-06-02 | Massachusetts Institute Of Technology | Cre-lox based gene knockdown constructs and methods of use thereof |
AU2008232316A1 (en) | 2007-03-26 | 2008-10-02 | Newcastle Innovation Limited | Therapeutic targets and molecules |
CA2687336C (en) | 2007-05-23 | 2014-12-23 | Dharmacon, Inc. | Micro-rna scaffolds and non-naturally occurring micro-rnas |
WO2009012263A2 (en) | 2007-07-18 | 2009-01-22 | The Trustees Of Columbia University In The City Of New York | Tissue-specific micrornas and compositions and uses thereof |
WO2009012468A2 (en) | 2007-07-18 | 2009-01-22 | The Regents Of The University Colorado | Differential expression of micrornas in nonfailing versus failing human hearts |
US20090092980A1 (en) | 2007-07-20 | 2009-04-09 | Christoph Arenz | miRNA PROCESSING INHIBITOR EFFICACY ASSAYS AND SUBSTANCES |
WO2009044895A1 (en) | 2007-10-03 | 2009-04-09 | Kyowa Hakko Kirin Co., Ltd. | Composition inhibiting the expression of target gene |
CA2704056A1 (en) | 2007-10-29 | 2009-05-07 | University Of Massachusetts | Encapsulated nanoparticles for nucleic acid delivery |
WO2009058818A2 (en) | 2007-10-29 | 2009-05-07 | The Board Of Regents Of The University Of Texas System | Compositions comprising a micro-rna and methods of their use in regulating cardiac remodeling |
US8404659B2 (en) | 2008-03-07 | 2013-03-26 | Santaris Pharma A/S | Pharmaceutical compositions for treatment of MicroRNA related diseases |
EP2279267A4 (en) | 2008-03-27 | 2012-01-18 | Vascular Biosciences Inc | Methods of novel therapeutic candidate identification through gene expression analysis in vascular-related diseases |
EP2105145A1 (en) | 2008-03-27 | 2009-09-30 | ETH Zürich | Method for muscle-specific delivery lipid-conjugated oligonucleotides |
US20090326049A1 (en) | 2008-04-04 | 2009-12-31 | Alexander Aristarkhov | Blocking oligos for inhibition of microrna and sirna activity and uses thereof |
US20100113284A1 (en) | 2008-04-04 | 2010-05-06 | Alexander Aristarkhov | Small interfering rna (sirna) target site blocking oligos and uses thereof |
EP2990487A1 (en) | 2008-05-08 | 2016-03-02 | Asuragen, INC. | Compositions and methods related to mirna modulation of neovascularization or angiogenesis |
CA2726052A1 (en) | 2008-06-04 | 2009-12-10 | The Board Of Regents Of The University Of Texas System | Modulation of gene expression through endogenous small rna targeting of gene promoters |
WO2010036939A2 (en) | 2008-09-26 | 2010-04-01 | University Of Southern California | A system for synergistic expression of multiple small functional rna elements |
EP2358398A2 (en) | 2008-10-24 | 2011-08-24 | Isis Pharmaceuticals, Inc. | Oligomeric compounds and methods |
WO2010126355A1 (en) | 2009-04-29 | 2010-11-04 | Academisch Medisch Centrum Bij De Universiteit Van Amsterdam | Means and methods for counteracting, preventing and/or determining heart failure, or a risk of heart failure |
CN102803284B (en) | 2009-06-08 | 2015-11-25 | 米拉根医疗公司 | For the chemically modified motif of miRNA inhibitor and stand-in |
-
2013
- 2013-06-19 US US13/921,537 patent/US9163235B2/en not_active Expired - Fee Related
- 2013-06-21 MD MDA20150006A patent/MD20150006A2/en not_active Application Discontinuation
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- 2013-06-21 KR KR1020157001535A patent/KR20150036140A/en not_active Application Discontinuation
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090175827A1 (en) * | 2006-12-29 | 2009-07-09 | Byrom Mike W | miR-16 REGULATED GENES AND PATHWAYS AS TARGETS FOR THERAPEUTIC INTERVENTION |
US20100298410A1 (en) * | 2007-10-04 | 2010-11-25 | Santaris Pharma A/S | MICROMIRs |
US20100317713A1 (en) * | 2007-11-09 | 2010-12-16 | Eric Olson | Micro-rnas of the mir-15 family modulate cardiomyocyte survival and cardiac repair |
Non-Patent Citations (2)
Title |
---|
HULLINGER, TG ET AL.: "Inhibition Of miR-15 Protects Against Cardiac Ischemic Injury", CIRCULATION RESEARCH, vol. 110, 3 November 2011 (2011-11-03), pages 71 - 81, XP055181810 * |
See also references of EP2863956A4 * |
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WO2016022536A2 (en) | 2014-08-04 | 2016-02-11 | MiRagen Therapeutics, Inc. | Inhibitors of myh7b and uses thereof |
US9487783B2 (en) | 2014-08-07 | 2016-11-08 | Regulus Therapeutics Inc. | Targeting microRNAs for metabolic disorders |
US9862950B2 (en) | 2014-08-07 | 2018-01-09 | Regulus Therapeutics Inc. | Targeting microRNAs for metabolic disorders |
US10138484B2 (en) | 2014-08-07 | 2018-11-27 | Regulus Therapeutics Inc. | Targeting microRNAs for metabolic disorders |
WO2017219170A1 (en) * | 2016-06-19 | 2017-12-28 | 毛侃琅 | Construction and application of lentiviral vector for specifically inhibiting human mirna-29a and mir-424 expression |
EP4035659A1 (en) | 2016-11-29 | 2022-08-03 | PureTech LYT, Inc. | Exosomes for delivery of therapeutic agents |
WO2021020412A1 (en) | 2019-07-30 | 2021-02-04 | 塩野義製薬株式会社 | Nucleic acid drug targeting murf1 |
WO2022018269A1 (en) * | 2020-07-23 | 2022-01-27 | Johann Wolfgang Goethe-Universität Frankfurt | Combinatorial inhibition of mirnas for treatment of heart failure |
Also Published As
Publication number | Publication date |
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CA2876105A1 (en) | 2013-12-27 |
KR20150036140A (en) | 2015-04-07 |
SG11201408460UA (en) | 2015-01-29 |
AR091539A1 (en) | 2015-02-11 |
HK1209621A1 (en) | 2016-04-08 |
BR112014032239A2 (en) | 2017-08-01 |
CN104540527A (en) | 2015-04-22 |
IN2014DN10899A (en) | 2015-09-11 |
AU2013277033A1 (en) | 2015-01-22 |
EP2863956A1 (en) | 2015-04-29 |
EP2863956A4 (en) | 2016-01-20 |
MD20150006A2 (en) | 2015-06-30 |
MX2014015641A (en) | 2015-07-14 |
US9163235B2 (en) | 2015-10-20 |
EA201590070A1 (en) | 2015-04-30 |
TW201406774A (en) | 2014-02-16 |
US20130345288A1 (en) | 2013-12-26 |
JP2015525081A (en) | 2015-09-03 |
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