THERAPEUTIC UNA OLIGOMERS AND USES THEREOF

Abstract

This invention provides UNA oligomers for regulating the expression of a target gene. The UNA oligomers contain UNA monomer linkers, and may contain one or more nucleotides modified with a 2-O-methyl group, one or more nucleotides modified with a 2-deoxy-2-fluoro group, and one or more phosphorothioate or chiral phosphorothioate intermonomer linkages. UNA oligomers can be used as active agents for preventing or treating disease.

Claims

1. A UNA oligomer comprising a first strand and a second strand, each of the strands being 19-29 monomers in length, the monomers comprising UNA monomers and nucleic acid monomers, wherein the oligomer has a duplex structure of from 14 to 29 monomers in length, wherein the second strand is a guide strand for RNA interference and the first strand is a passenger strand for RNA interference, and wherein the UNA oligomer reduces expression of a target gene.

2. The UNA oligomer of claim 1, comprising one or more chemical modifications of one or more of the nucleic acid monomers in each strand.

3. The UNA oligomer of claim 1, wherein the oligomer contains one or more nucleotides modified with a 2-O-methyl group.

4. The UNA oligomer of claim 1, wherein the oligomer contains from one to nine nucleotides modified with a 2-O-methyl group.

5. The UNA oligomer of claim 1, wherein the oligomer contains one or more nucleotides modified with a 2-deoxy-2-fluoro group.

6. The UNA oligomer of claim 1, wherein one or more of the intermonomer linkages in either strand is a phosphorothioate or chiral phosphorothioate linkage.

7. The UNA oligomer of claim 1, wherein the oligomer contains one or more nucleotides modified with a 2-O-methyl group and one or more nucleotides modified with a 2-deoxy-2-fluoro group.

8. The UNA oligomer of claim 1, wherein the oligomer contains one or more nucleotides modified with a 2-O-methyl group, one or more nucleotides modified with a 2-deoxy-2-fluoro group, and one or more phosphorothioate or chiral phosphorothioate intermonomer linkages in either strand.

9. The UNA oligomer of claim 1, wherein the oligomer contains three or more UNA monomers, one or more nucleotides modified with a 2-O-methyl group, one or more nucleotides modified with a 2-deoxy-2-fluoro group, and one or more phosphorothioate or chiral phosphorothioate intermonomer linkages in either strand.

10. The UNA oligomer of claim 1, wherein the target gene is a viral gene of Hepatitis B virus, TTR, APOB, PCSK9, or APOC3.

11. The UNA oligomer of claim 1, comprising a first strand being SEQ ID NO:87 and a second strand being SEQ ID NO:88.

12. The UNA oligomer of claim 1, comprising a first strand being SEQ ID NO:105 and a second strand being SEQ ID NO:106.

13. The UNA oligomer of claim 1, comprising a first strand being SEQ ID NO:239 and a second strand being SEQ ID NO:240.

14. The UNA oligomer of claim 1, comprising a first strand being SEQ ID NO:263 and a second strand being SEQ ID NO:264.

15. The UNA oligomer of claim 1, wherein the UNA oligomer has long lasting activity in vitro.

16. The UNA oligomer of claim 1, wherein the UNA oligomer has long lasting potency in vivo.

17. A pharmaceutical composition comprising one or more UNA oligomers of any of claims 1-16 and a pharmaceutically acceptable carrier, diluent, or adjuvant.

18. Use of one or more UNA oligomers of any of claims 1-16 to regulate expression of a target gene.

19. Use of the composition of claim 17 in preventing or treating Hepatitis B infection, amyloid neuropathy, amyloidosis, amyloidosis related to transthyretin, hypercholesterolemia, cholesterol disorder, hypertriglyceridemia, or lipoprotein disorder in a subject in need thereof.

20. Use of the composition of claim 17 in the preparation of a medicament for a therapeutic target.

21. Use of one or more UNA oligomers of any of claims 1-16 in the preparation of a medicament for decreasing expression level of a target mRNA in a cell.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0029] This invention provides oligomer structures for therapeutic agents that are long acting in their effect on gene regulation when used in vitro or administered in vivo. The oligomeric structures of this invention contain one or more UNA monomers.

[0030] This invention provides structures, methods and compositions for UNA monomer-containing oligomeric agents that incorporate UNA monomers. The UNA oligomeric molecules of this invention can be used as active agents in formulations for gene silencing therapeutics.

[0031] Among other things, this invention provides unique oligomer structures that incorporate novel combinations of UNA monomers with certain natural nucleotides, non-natural nucleotides, modified nucleotides, or chemically-modified nucleotides.

[0032] Aspects of this disclosure include UNA monomer-containing oligomers that are pharmacologically active molecules. The UNA Oligomers of this invention can be used as active pharmaceutical ingredients for regulating or modulating gene expression, RNA interference methods, as well as antisense strategies.

[0033] Embodiments of this disclosure provide a wide range of novel UNA oligomer molecules, which incorporate one or more UNA monomer linkers.

[0034] In some aspects, this invention provides long acting UNA Oligomers having structures that incorporate novel combinations of UNA Monomers with certain natural nucleotides, non-natural nucleotides, modified nucleotides, or chemically-modified nucleotides.

[0035] In some aspects, this invention provides long acting properties for gene silencing drugs, and can reduce the dose levels required for efficacious therapy.

[0036] In certain aspects, this invention can provide UNA oligomers having increased activity in vitro.

[0037] In some embodiments, this invention can provide UNA oligomers having increased potency in vivo.

[0038] In additional embodiments, this invention can provide UNA oligomers having longer functional half-life.

[0039] In certain embodiments, this invention can provide UNA oligomers having long lasting activity in vitro.

[0040] In particular embodiments, this invention can provide UNA oligomers having long lasting potency in vivo.

[0041] Embodiments of this invention may further provide UNA oligomers having increased enzymatic stability.

[0042] UNA oligomers of this disclosure can provide enhanced stability, in vitro and in vivo, for modulating gene expression.

[0043] In further aspects of this invention, UNA oligomers can exhibit reduced side effects as agents for treatment of a disease or condition. In some embodiments, a UNA oligomer can exhibit reduced off target effects as an agent for treating a disease, or in vitro.

[0044] UNA Monomers

[0045] UNA monomers are small organic molecules based on a propane-1,2,3-tri-yl-trisoxy structure as shown below:

##STR00001##

where R.sup.1 and R.sup.2 are H, and R.sup.1 and R.sup.2 can be phosphodiester linkages, Base can be a nucleobase, and R.sup.3 is a functional group described below.

[0046] In another view, the UNA monomer main atoms can be drawn in IUPAC notation as follows:

##STR00002##

where the direction of progress of the oligomer chain is from the 1-end to the 3-end of the propane residue.

[0047] Examples of a nucleobase include uracil, thymine, cytosine, 5-methylcytosine, adenine, guanine, inosine, and natural and non-natural nucleobase analogues.

[0048] In general, because the UNA monomers are not nucleotides, they can exhibit at least four forms in an oligomer. First, a UNA monomer can be an internal monomer in an oligomer, where the UNA monomer is flanked by other monomers on both sides. In this form, the UNA monomer can participate in base pairing when the oligomer is a duplex, for example, and there are other monomers with nucleobases in the duplex.

[0049] Examples of UNA monomer as internal monomers flanked at both the propane-1-yl position and the propane-3-yl position, where R.sup.3 is OH, are shown below.

##STR00003## ##STR00004##

[0050] Second, a UNA monomer can be a monomer in an overhang of an oligomer duplex, where the UNA monomer is flanked by other monomers on both sides. In this form, the UNA monomer does not participate in base pairing. Because the UNA monomers are flexible organic structures, unlike nucleotides, the overhang containing a UNA monomer will be a flexible terminator for the oligomer.

[0051] A UNA monomer can be a terminal monomer in an overhang of an oligomer, where the UNA monomer is attached to only one monomer at either the propane-1-yl position or the propane-3-yl position. In this form, the UNA monomer does not participate in base pairing. Because the UNA monomers are flexible organic structures, unlike nucleotides, the overhang containing a UNA monomer can be a flexible terminator for the oligomer.

[0052] Examples of a UNA monomer as a terminal monomer attached at the propane-3-yl position are shown below.

##STR00005##

[0053] Because a UNA monomer can be a flexible molecule, a UNA monomer as a terminal monomer can assume widely differing conformations. An example of an energy minimized UNA monomer conformation as a terminal monomer attached at the propane-3-yl position is shown below.

##STR00006##

Thus, UNA oligomers having a terminal UNA monomer are significantly different in structure from conventional nucleic acid agents, such as siRNAs. For example, siRNAs may require that terminal monomers or overhangs in a duplex be stabilized. In contrast, the conformability of a terminal UNA monomer can provide UNA oligomers with different properties.

[0054] Among other things, the structure of the UNA monomer allows it to be attached to naturally-occurring nucleotides. A UNA oligomer can be a chain composed of UNA monomers, as well as various nucleotides that may be based on naturally-occurring nucleosides.

[0055] In some embodiments, the functional group R.sup.3 of a UNA monomer can be OR.sup.4, SR.sup.4, NR.sup.4.sub.2, NH(CO)R.sup.4, morpholino, morpholin-1-yl, piperazin-1-yl, or 4-alkanoyl-piperazin-1-yl, where R.sup.4 is the same or different for each occurrence, and can be H, alkyl, a cholesterol, a lipid molecule, a polyamine, an amino acid, or a polypeptide.

[0056] The UNA monomers are organic molecules. UNA monomers are not nucleic acid monomers or nucleotides, nor are they naturally-occurring nucleosides or modified naturally-occurring nucleosides.

[0057] A UNA oligomer of this invention is a synthetic chain molecule. A UNA oligomer of this invention is not a nucleic acid, nor an oligonucleotide.

[0058] In some embodiments, as shown above, a UNA monomer can be UNA-A (designated ), UNA-U (designated ), UNA-C (designated ), and UNA-G (designated ).

[0059] Designations that may be used herein include mA, mG, mC, and mU, which refer to the 2-O-methyl modified ribonucleotides.

[0060] Designations that may be used herein include lower case c and u, which refer to the 2-O-methyl modified ribonucleotides.

[0061] Designations that may be used herein include dT, which refers to a 2-deoxy T nucleotide.

[0062] Designations that may be used herein include *, which refers to a phosphorothioate linkage.

[0063] Monomers for UNA Oligomers

[0064] As used herein, in the context of oligomer sequences, the symbol X represents a UNA monomer.

[0065] As used herein, in the context of oligomer sequences, the symbol N represents any natural nucleotide monomer, or a modified nucleotide monomer.

[0066] As used herein, in the context of oligomer sequences, the symbol Q represents a non-natural, modified, or chemically-modified nucleotide monomer.

[0067] When a Q monomer appears in one strand of the oligomer, and is unpaired with the other strand, the monomer can have any base attached. When a Q monomer appears in one strand of the oligomer, and is paired with a monomer in the other strand, the Q monomer can have any base attached that would be complementary to the monomer in the corresponding paired position in the other strand.

[0068] Examples of nucleic acid monomers include non-natural, modified, and chemically-modified nucleotides, including any such nucleotides known in the art.

[0069] Examples of non-natural, modified, and chemically-modified nucleotide monomers include 2-O-methyl ribonucleotides, 2-O-methyl purine nucleotides, 2-deoxy-2-fluoro ribonucleotides, 2-deoxy-2-fluoro pyrimidine nucleotides, 2-deoxy ribonucleotides, 2-deoxy purine nucleotides, universal base nucleotides, 5-C-methyl-nucleotides, and inverted deoxyabasic monomer residues.

[0070] Examples of non-natural, modified, and chemically-modified nucleotide monomers include 3-end stabilized nucleotides, 3-glyceryl nucleotides, 3-inverted abasic nucleotides, and 3-inverted thymidine.

[0071] Examples of non-natural, modified, and chemically-modified nucleotide monomers include locked nucleic acid nucleotides (LNA), 2-0,4-C-methylene-(D-ribofuranosyl) nucleotides, 2-methoxyethoxy (MOE) nucleotides, 2-methyl-thio-ethyl, 2-deoxy-2-fluoro nucleotides, and 2-O-methyl nucleotides.

[0072] Examples of non-natural, modified, and chemically-modified nucleotide monomers include 2,4-Constrained 2-O-Methoxyethyl (cMOE) and 2-O-Ethyl (cEt) modified DNAs.

[0073] Examples of non-natural, modified, and chemically-modified nucleotide monomers include 2-amino nucleotides, 2-O-amino nucleotides, 2-C-allyl nucleotides, and 2-O-allyl nucleotides.

[0074] Examples of non-natural, modified, and chemically-modified nucleotide monomers include N.sup.6-methyladenosine nucleotides.

[0075] Examples of non-natural, modified, and chemically-modified nucleotide monomers include nucleotide monomers with modified bases 5-(3-amino)propyluridine, 5-(2-mercapto)ethyluridine, 5-bromouridine; 8-bromoguanosine, or 7-deazaadenosine.

[0076] Examples of non-natural, modified, and chemically-modified nucleotide monomers include 2-O-aminopropyl substituted nucleotides.

[0077] Examples of non-natural, modified, and chemically-modified nucleotide monomers include replacing the 2-OH group of a nucleotide with a 2-R, a 2-OR, a 2-halogen, a 2-SR, or a 2-amino, where R can be H, alkyl, alkenyl, or alkynyl.

[0078] Some examples of modified nucleotides are given in Saenger, Principles of Nucleic Acid Structure, Springer-Verlag, 1984.

[0079] UNA Oligomeric Compounds Containing UNA Monomer Linkers

[0080] Aspects of this invention can provide structures and compositions for UNA-containing oligomeric compounds. The oligomeric agents may incorporate one or more UNA monomers. Oligomeric molecules of this invention can be used as active agents in formulations for gene regulating or gene silencing therapeutics.

[0081] An oligomer can be single stranded, or double stranded, or may have additional strands or non-strand structures.

[0082] In some embodiments, this invention provides oligomeric compounds having a structure that incorporates novel combinations of UNA monomers with certain natural nucleotides, non-natural nucleotides, modified nucleotides, or chemically-modified nucleotides.

[0083] In further aspects, the oligomeric compounds can be pharmacologically active molecules. A UNA oligomer of this invention can be used as an active pharmaceutical ingredient for regulating or modulating gene expression, and in RNA interference methods, as well as antisense, RNA blocking, and micro-RNA strategies.

[0084] A UNA oligomer of this invention can have the structure of Formula I

##STR00007##

wherein L.sup.1 is a linkage, n is from 19 to 29, and for each occurrence L.sup.2 is a UNA linker group having the formula C.sup.1C.sup.2C.sup.3, where R is attached to C.sup.2 and has the formula OCH(CH.sub.2R.sup.3)R.sup.5, where R.sup.3 is OR.sup.4, SR.sup.4, NR.sup.4.sub.2, NH(CO)R.sup.4, morpholino, morpholin-1-yl, piperazin-1-yl, or 4-alkanoyl-piperazin-1-yl, where R.sup.4 is the same or different for each occurrence and is H, alkyl, a cholesterol, a lipid molecule, a polyamine, an amino acid, or a polypeptide, and where R.sup.5 is a nucleobase, or L.sup.2(R) is a sugar such as a ribose and R is a nucleobase, or L.sup.2 is a modified sugar such as a modified ribose and R is a nucleobase. In certain embodiments, a nucleobase can be a modified nucleobase. L.sup.1 can be a phosphodiester linkage.

[0085] A UNA oligomer of this invention can be a short chain molecule. A UNA oligomer can be a duplex pair. Thus, a UNA oligomer can have a first strand of the duplex and a second strand of the duplex, which is complementary to the first strand, with respect to the nucleobases, although up to three mismatches can occur. A UNA oligomer duplex can have overhangs.

[0086] In some embodiments, a 3 overhang can be present on both strands, which can be 2 monomers in length. For example, a 3 overhang can be TT or uu.

[0087] Some UNA oligomers are discussed in U.S. Pat. No. 8,314,227, as well as US Patent Publication No. 20110313020 A1.

[0088] The target of a UNA oligomer can be a target nucleic acid. In some embodiments, the target can be any mRNA of a subject. A UNA oligomer can be active for gene silencing in RNA interference.

[0089] A UNA oligomer may comprise two strands that together provide a duplex. The duplex may be composed of a first strand, which may also be referred to as a passenger strand or sense strand, and a second strand, which may also be referred to as a guide strand or antisense strand.

[0090] In some aspects, a UNA oligomer of this invention can have any number of phosphorothioate intermonomer linkages in any position in any strand, or in both strands of a duplex structure.

[0091] In some embodiments, any one or more of the intermonomer linkages of a UNA oligomer can be a phosphodiester, a phosphorothioate including dithioates, a chiral phosphorothioate, and other chemically modified forms.

[0092] Examples of UNA oligomers of this invention include duplex pairs, which are in general complementary. Thus, for example, SEQ ID NO:1 can represent a first strand of a duplex and SEQ ID NO:2 can represent a second strand of the duplex, which is complementary to the first strand.

[0093] For example, the symbol N in the first strand can represent any nucleotide that is complementary to the monomer in the corresponding position in the second strand. Example UNA oligomers of this disclosure are shown with 2-monomer length overhangs, although overhangs of from 1 to 8 monomers, or longer, can be used.

[0094] The symbol X in a strand or oligomer represents a UNA monomer.

[0095] When a UNA monomer appears in one strand of the oligomer, and is unpaired with the other strand, the monomer can have any base attached. When a UNA monomer appears in one strand of the oligomer, and is paired with a monomer in the other strand, the UNA monomer can have any base attached that would be complementary to the monomer in the corresponding paired position in the other strand.

[0096] Further, when the oligomer terminates in a UNA monomer, the terminal position has a 1-end, according to the positional numbering shown above, instead of a 5-end as for a nucleotide, or the terminal position has a 3-end, according to the positional numbering shown above, instead of a 3-end as for a nucleotide. For example, the UNA oligomer

TABLE-US-00001 SEQIDNO:1 1-X.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N-X.Math.X-3 SEQIDNO:2 3-X.Math.X.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.N.Math.X.Math.X.Math.X.Math.X.Math.X.Math.X.Math.X.Math.N-5
has a UNA monomer 1-end on the first strand, a UNA monomer 3-end on the first strand, a UNA monomer 3-end on the second strand, and a nucleotide 5-end on the second strand.

[0097] In some embodiments, a UNA oligomer of this invention can have one or more UNA monomers at the 1-end of the first strand, and one or more UNA monomers at the 3-end of the first strand.

[0098] In further embodiments, a UNA oligomer of this invention can have one or more UNA monomers at the 3-end of the second strand.

[0099] In certain embodiments, a duplex UNA oligomer of this invention can have one or more UNA monomers at the 1-end of the first strand, one or more UNA monomers at the 3-end of the first strand, and one or more UNA monomers at the 3-end of the second strand.

[0100] A UNA oligomer of this invention the oligomer may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length.

[0101] In certain embodiments, a UNA oligomer of this invention may have a first strand that is 19-23 monomers in length.

[0102] In certain embodiments, a UNA oligomer of this invention may have a duplex region that is 19-21 monomers in length.

[0103] In further embodiments, a UNA oligomer of this invention may have a second strand that is 19-23 monomers in length.

[0104] In certain embodiments, a UNA oligomer of this invention may have a first strand that is 19 monomers in length, and a second strand that is 21 monomers in length.

[0105] In certain embodiments, a UNA oligomer of this invention may have a first strand that is 20 monomers in length, and a second strand that is 21 monomers in length.

[0106] In certain embodiments, a UNA oligomer of this invention may have a first strand that is 21 monomers in length, and a second strand that is 21 monomers in length.

[0107] In certain embodiments, a UNA oligomer of this invention may have a first strand that is 22 monomers in length, and a second strand that is 21 monomers in length.

[0108] A UNA oligomer of this invention for inhibiting gene expression can have a first strand and a second strand, each of the strands being 19-29 monomers in length. The monomers can be UNA monomers and nucleic acid monomers. The oligomer can have a duplex structure of from 14 to 29 monomers in length. The UNA oligomer can be targeted to a target gene and can exhibit reduced off-target effects as compared to a conventional siRNA. In some embodiments, a UNA oligomer of this invention can have a first strand and a second strand, each of the strands being 19-23 monomers in length.

[0109] In another aspect, the UNA oligomer may have a blunt end, or may have one or more overhangs. In some embodiments, the first and second strands may be connected with a connecting oligomer in between the strands, and form a duplex region with a connecting loop at one end.

[0110] In certain embodiments, an overhang can be one or two monomers in length.

[0111] A UNA oligomer can mediate cleavage of a target nucleic acid in a cell. In some processes, the second strand of the UNA oligomer, at least a portion of which can be complementary to the target nucleic acid, can act as a guide strand that can hybridize to the target nucleic acid.

[0112] The second strand can be incorporated into an RNA Induced Silencing Complex (RISC).

[0113] A UNA oligomer of this disclosure may comprise naturally-occurring nucleic acid nucleotides, and modifications thereof that are compatible with gene silencing activity.

[0114] In some aspects, a UNA oligomer is a double stranded construct molecule that is able to inhibit gene expression.

[0115] As used herein, the term strand refers to a single, contiguous chain of monomers, the chain having any number of internal monomers and two end monomers, where each end monomer is attached to one internal monomer on one side, and is not attached to a monomer on the other side, so that it ends the chain.

[0116] The monomers of a UNA oligomer may be attached via phosphodiester linkages, phosphorothioate linkages, gapped linkages, and other variations.

[0117] In some embodiments, a UNA oligomer can include mismatches in complementarity between the first and second strands. In other embodiments, a UNA oligomer may have 1, or 2, or 3 mismatches. The mismatches may occur at any position in the duplex region.

[0118] The target of a UNA oligomer can be a target nucleic acid of a target gene.

[0119] A UNA oligomer may have one or two overhangs outside the duplex region. The overhangs can be an unpaired portion at the end of the first strand or second strand. The lengths of the overhang portions of the first and second strands can be the same or different.

[0120] A UNA oligomer may have at least one blunt end. A blunt end does not have an overhang portion, and the duplex region at a blunt end terminates at the same position for both the first and second strands.

[0121] A UNA oligomer can be RISC length, which means that it has a duplex length of less than 25 base pairs.

[0122] In certain embodiments, a UNA oligomer can be a single strand that folds upon itself and hybridizes to itself to form a double stranded region having a connecting loop at the end of the double stranded region.

[0123] Examples of UNA oligomers containing five UNA Monomers, and which may contain one or more Q monomers are shown in Table 1.

TABLE-US-00002 TABLE1 5XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 3 XQNNQNQNQQQNQNQNQNQNXX 4 XXQNQNQNNNNNQNQNQNQNQ 5 XQNNQNQNQNQNQNQNQNQNXX 6 XXQNQNQNQNQNQNQNQNQNQ 7 XQNNQNQNNNNNQNQNQNQNXX 8 XXQNQNQNQNNNQNQNQNQNQ 9 XQNNQNQNNNNNQNQNQNQNXX 10 XXQNQNQNNNNNNNQNQNQNQ 11 XQNNQNQNNNNNNNQNQNQNXX 12 XXQNQNQNNNNNNNNNQNQNQ 13 XQNNQNQNNNNNNNNNQNQNXX 14 XXQNQNQNNNNNNNNNQNQNQ 15 XQNNQNQNNNNNNNNNQNQNXX 16 XXQNQNQNNNNNNNNNNNQNQ 17 XQNNQNNNNNNNNNNNQNQNXX 18 XXQNQNQNNNNNNNNNNNQNQ 19 XQNNQNNNNNNNNNNNQNQNXX 20 XXQNQNNNNNNNNNNNNNQNQ 21 XQNNNNNNNNNNNNNNQNQNXX 22 XXQNQNNNNNNNNNNNNNQNQ 23 XQNNNNNNNNNNNNNNNNQNXX 24 XXQNQNNNNNNNNNNNNNQNQ 25 XQNNNNNNNNNNNNNNNNQNXX 26 XXQNNNNNNNNNNNNNNNQNQ 27 XQNNNNNNNNNNNNNNNNQNXX 28 XXQNNNNNNNNNNNNNNNNNQ 29 XQNNNNNNNNNNNNNNNNNNXX 30 XXQNNNNNNNNNNNNNNNNNQ

[0124] Examples of UNA oligomers containing four UNA Monomers that are enriched in Q monomers are shown in Table 2.

TABLE-US-00003 TABLE2 4XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 31 XQNNQNQNQQQNQNQNQNQNXQ 32 XXQNQNQNNNNNQNQNQNQNQ 33 XQNNQNQNQNQNQNQNQNQNXQ 34 XXQNQNQNQNQNQNQNQNQNQ 35 XQNNQNQNNNNNQNQNQNQNXQ 36 XXQNQNQNQNNNQNQNQNQNQ 37 XQNNQNQNNNNNQNQNQNQNXQ 38 XXQNQNQNNNNNNNQNQNQNQ 39 XQNNQNQNNNNNNNQNQNQNXQ 40 XXQNQNQNNNNNNNNNQNQNQ 41 XQNNQNQNNNNNNNNNQNQNXQ 42 XXQNQNQNNNNNNNNNQNQNQ 43 XQNNQNQNNNNNNNNNQNQNXQ 44 XXQNQNQNNNNNNNNNNNQNQ 45 XQNNQNNNNNNNNNNNQNQNXQ 46 XXQNQNQNNNNNNNNNNNQNQ 47 XQNNQNNNNNNNNNNNQNQNXQ 48 XXQNQNNNNNNNNNNNNNQNQ 49 XQNNNNNNNNNNNNNNQNQNXQ 50 XXQNQNNNNNNNNNNNNNQNQ 51 XQNNNNNNNNNNNNNNNNQNXQ 52 XXQNQNNNNNNNNNNNNNQNQ 53 XQNNNNNNNNNNNNNNNNQNXQ 54 XXQNNNNNNNNNNNNNNNQNQ 55 XQNNNNNNNNNNNNNNNNQNXQ 56 XXQNNNNNNNNNNNNNNNNNQ 57 XQNNNNNNNNNNNNNNNNNNXQ 58 XXQNNNNNNNNNNNNNNNNNQ

[0125] Examples of UNA oligomers containing four UNA Monomers that are enriched in Q monomers are shown in Table 3.

TABLE-US-00004 TABLE3 4XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 59 XQNQNQNQQQNQNQNQNQNXX 60 QXQNQNQNNNNNQNQNQNQNQ 61 XQNQNQNQNQNQNQNQNQNXX 62 QXQNQNQNQNQNQNQNQNQNQ 63 XQNQNQNNNNNQNQNQNQNXX 64 QXQNQNQNQNNNQNQNQNQNQ 65 XQNQNQNNNNNQNQNQNQNXX 66 QXQNQNQNNNNNNNQNQNQNQ 67 XQNQNQNNNNNNNQNQNQNXX 68 QXQNQNQNNNNNNNNNQNQNQ 69 XQNQNQNNNNNNNNNQNQNXX 70 QXQNQNQNNNNNNNNNQNQNQ 71 XQNQNQNNNNNNNNNQNQNXX 72 QXQNQNQNNNNNNNNNNNQNQ 73 XQNQNNNNNNNNNNNQNQNXX 74 QXQNQNQNNNNNNNNNNNQNQ 75 XQNQNNNNNNNNNNNQNQNXX 76 QXQNQNNNNNNNNNNNNNQNQ 77 XQNNNNNNNNNNNNNQNQNXX 78 QXQNQNNNNNNNNNNNNNQNQ 79 XQNNNNNNNNNNNNNNNQNXX 80 QXQNQNNNNNNNNNNNNNQNQ 81 XQNNNNNNNNNNNNNNNQNXX 82 QXQNNNNNNNNNNNNNNNQNQ 83 XQNNNNNNNNNNNNNNNQNXX 84 QXQNNNNNNNNNNNNNNNNNQ 85 XQNNNNNNNNNNNNNNNNNXX 86 QXQNNNNNNNNNNNNNNNNNQ

[0126] Examples of UNA oligomers containing three UNA Monomers that are enriched in Q monomers are shown in Table 4.

TABLE-US-00005 TABLE4 3XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 87 XQNQNQNQQQNQNQNQNQNXQ 88 QXQNQNQNNNNNQNQNQNQNQ 89 XQNQNQNQNQNQNQNQNQNXQ 90 QXQNQNQNQNQNQNQNQNQNQ 91 XQNQNQNNNNNQNQNQNQNXQ 92 QXQNQNQNQNNNQNQNQNQNQ 93 XQNQNQNNNNNQNQNQNQNXQ 94 QXQNQNQNNNNNNNQNQNQNQ 95 XQNQNQNNNNNNNQNQNQNXQ 96 QXQNQNQNNNNNNNNNQNQNQ 97 XQNQNQNNNNNNNNNQNQNXQ 98 QXQNQNQNNNNNNNNNQNQNQ 99 XQNQNQNNNNNNNNNQNQNXQ 100 QXQNQNQNNNNNNNNNNNQNQ 101 XQNQNNNNNNNNNNNQNQNXQ 102 QXQNQNQNNNNNNNNNNNQNQ 103 XQNQNNNNNNNNNNNQNQNXQ 104 QXQNQNNNNNNNNNNNNNQNQ 105 XQNNNNNNNNNNNNNQNQNXQ 106 QXQNQNNNNNNNNNNNNNQNQ 107 XQNNNNNNNNNNNNNNNQNXQ 108 QXQNQNNNNNNNNNNNNNQNQ 109 XQNNNNNNNNNNNNNNNQNXQ 110 QXQNNNNNNNNNNNNNNNQNQ 111 XQNNNNNNNNNNNNNNNQNXQ 112 QXQNNNNNNNNNNNNNNNNNQ 113 XQNNNNNNNNNNNNNNNNNXQ 114 QXQNNNNNNNNNNNNNNNNNQ

[0127] Examples of UNA oligomers containing six UNA Monomers that are enriched in Q monomers are shown in Table 5.

TABLE-US-00006 TABLE5 6XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 115 XQNQNQNQQQNQNQNQNQNXX 116 XXQNQNQNNNNNQNQNQNQXQ 117 XQNQNQNQQQNQNQNQNQNXX 118 XXQNQNQNNNNNQNQNQNXNQ 119 XQNQNQNQQQNQNQNQNQNXX 120 XXQNQNQNNNNNQNQNQXQNQ 121 XQNQNQNQQQNQNQNQNQNXX 122 XXQNQNQNNNNNQNQNXNQNQ 123 XQNQNQNQQQNQNQNQNQNXX 124 XXQNQNQNNNNNQNQXQNQNQ 125 XQNQNQNQQQNQNQNQNQNXX 126 XXQNQNQNNNNNQNXNQNQNQ 127 XQNQNQNQQQNQNQNQNQNXX 128 XXQNQNQNNNNNQXQNQNQNQ

[0128] Examples of UNA oligomers containing seven UNA Monomers that are enriched in Q monomers are shown in Table 6.

TABLE-US-00007 TABLE6 7XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 129 XQNQNQNQQQNQNQNQNQNXX 130 XXQNQNQNNNNNQNQNQNXXQ 131 XQNQNQNQQQNQNQNQNQNXX 132 XXQNQNQNNNNNQNQNQXQXQ 133 XQNQNQNQQQNQNQNQNQNXX 134 XXQNQNQNNNNNQNQNXNQXQ 135 XQNQNQNQQQNQNQNQNQNXX 136 XXQNQNQNNNNNQNQXQNXNQ 137 XQNQNQNQQQNQNQNQNQNXX 138 XXQNQNQNNNNNQNQNXXQNQ 139 XQNQNQNQQQNQNQNQNQNXX 140 XXQNQNQNNNNNQNXNQXQNQ 141 XQNQNQNQQQNQNQNQNQNXX 142 XXQNQNQNNNNNQNQXXNQNQ

[0129] Examples of UNA oligomers containing five UNA Monomers that are enriched in Q monomers are shown in Table 7.

TABLE-US-00008 TABLE7 5XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 143 XQNQNQNQQQNQNQNQNQNXQ 144 XXQNQNQNNNNNQNQNQNQXQ 145 XQNQNQNQQQNQNQNQNQNXQ 146 XXQNQNQNNNNNQNQNQNXNQ 147 XQNQNQNQQQNQNQNQNQNXQ 148 XXQNQNQNNNNNQNQNQXQNQ 149 XQNQNQNQQQNQNQNQNQNXQ 150 XXQNQNQNNNNNQNQNXNQNQ 151 XQNQNQNQQQNQNQNQNQNXQ 152 XXQNQNQNNNNNQNQXQNQNQ 153 XQNQNQNQQQNQNQNQNQNXQ 154 XXQNQNQNNNNNQNXNQNQNQ 155 XQNQNQNQQQNQNQNQNQNXQ 156 XXQNQNQNNNNNQXQNQNQNQ

[0130] Examples of UNA oligomers containing six UNA Monomers that are enriched in Q monomers are shown in Table 8.

TABLE-US-00009 TABLE8 6XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 157 XQNQNQNQQQNQNQNQNQNXQ 158 XXQNQNQNNNNNQNQNQNXXQ 159 XQNQNQNQQQNQNQNQNQNXQ 160 XXQNQNQNNNNNQNQNQXQXQ 161 XQNQNQNQQQNQNQNQNQNXQ 162 XXQNQNQNNNNNQNQNXNQXQ 163 XQNQNQNQQQNQNQNQNQNXQ 164 XXQNQNQNNNNNQNQXQNXNQ 165 XQNQNQNQQQNQNQNQNQNXQ 166 XXQNQNQNNNNNQNQNXXQNQ 167 XQNQNQNQQQNQNQNQNQNXQ 168 XXQNQNQNNNNNQNXNQXQNQ 169 XQNQNQNQQQNQNQNQNQNXQ 170 XXQNQNQNNNNNQNQXXNQNQ

[0131] Examples of UNA oligomers containing five UNA Monomers that are enriched in Q monomers are shown in Table 9.

TABLE-US-00010 TABLE9 5XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 171 XQNQNQNQQQNQNQNQNQNXX 172 QXQNQNQNNNNNQNQNQNQXQ 173 XQNQNQNQQQNQNQNQNQNXX 174 QXQNQNQNNNNNQNQNQNXNQ 175 XQNQNQNQQQNQNQNQNQNXX 176 QXQNQNQNNNNNQNQNQXQNQ 177 XQNQNQNQQQNQNQNQNQNXX 178 QXQNQNQNNNNNQNQNXNQNQ 179 XQNQNQNQQQNQNQNQNQNXX 180 QXQNQNQNNNNNQNQXQNQNQ 181 XQNQNQNQQQNQNQNQNQNXX 182 QXQNQNQNNNNNQNXNQNQNQ 183 XQNQNQNQQQNQNQNQNQNXX 184 QXQNQNQNNNNNQXQNQNQNQ

[0132] Examples of UNA oligomers containing six UNA Monomers that are enriched in Q monomers are shown in Table 10.

TABLE-US-00011 TABLE10 6XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 185 XQNQNQNQQQNQNQNQNQNXX 186 QXQNQNQNNNNNQNQNQNXXQ 187 XQNQNQNQQQNQNQNQNQNXX 188 QXQNQNQNNNNNQNQNQXQXQ 189 XQNQNQNQQQNQNQNQNQNXX 190 QXQNQNQNNNNNQNQNXNQXQ 191 XQNQNQNQQQNQNQNQNQNXX 192 QXQNQNQNNNNNQNQXQNXNQ 193 XQNQNQNQQQNQNQNQNQNXX 194 QXQNQNQNNNNNQNQNXXQNQ 195 XQNQNQNQQQNQNQNQNQNXX 196 QXQNQNQNNNNNQNXNQXQNQ 197 XQNQNQNQQQNQNQNQNQNXX 198 QXQNQNQNNNNNQNQXXNQNQ

[0133] Examples of UNA oligomers containing four UNA Monomers that are enriched in Q monomers are shown in Table 11.

TABLE-US-00012 TABLE11 4XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 199 XQNQNQNQQQNQNQNQNQNXQ 200 QXQNQNQNNNNNQNQNQNQXQ 201 XQNQNQNQQQNQNQNQNQNXQ 202 QXQNQNQNNNNNQNQNQNXNQ 203 XQNQNQNQQQNQNQNQNQNXQ 204 QXQNQNQNNNNNQNQNQXQNQ 205 XQNQNQNQQQNQNQNQNQNXQ 206 QXQNQNQNNNNNQNQNXNQNQ 207 XQNQNQNQQQNQNQNQNQNXQ 208 QXQNQNQNNNNNQNQXQNQNQ 209 XQNQNQNQQQNQNQNQNQNXQ 210 QXQNQNQNNNNNQNXNQNQNQ 211 XQNQNQNQQQNQNQNQNQNXQ 212 QXQNQNQNNNNNQXQNQNQNQ

[0134] Examples of UNA Oligomers containing five UNA Monomers that are enriched in Q monomers are shown in Table 12.

TABLE-US-00013 TABLE12 5XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 213 XQNQNQNQQQNQNQNQNQNXQ 214 QXQNQNQNNNNNQNQNQNXXQ 215 XQNQNQNQQQNQNQNQNQNXQ 216 QXQNQNQNNNNNQNQNQXQXQ 217 XQNQNQNQQQNQNQNQNQNXQ 218 QXQNQNQNNNNNQNQNXNQXQ 219 XQNQNQNQQQNQNQNQNQNXQ 220 QXQNQNQNNNNNQNQXQNXNQ 221 XQNQNQNQQQNQNQNQNQNXQ 222 QXQNQNQNNNNNQNQNXXQNQ 223 XQNQNQNQQQNQNQNQNQNXQ 224 QXQNQNQNNNNNQNXNQXQNQ 225 XQNQNQNQQQNQNQNQNQNXQ 226 QXQNQNQNNNNNQNQXXNQNQ

[0135] Examples of UNA oligomers containing seven or more UNA Monomers that are enriched in Q monomers are shown in Table 13.

TABLE-US-00014 TABLE13 7X-11XUNAoligomersenrichedinQmonomers SEQ ID NO: OLIGOMER 227 XQNQNQNQQQNQNQNQNQNXX 228 XXQNQNQNNNNNQNQXQXQXQ 229 XQNQNQNQQQNQNQNQNQNXQ 230 XXQNQNQNNNNNQNQNQXXXQ 231 XQNQNQNQQQNQNQNQNQNXX 232 QXQNQNQNNNNNQNXXXNQNQ 233 XQNQNQNQQQNQNQNQNQNXQ 234 QXQNQNQNNNNNQNXXXXXXQ 235 XQNQNQNQQQNQNQNQNQNXX 236 XXQNQNQNNNNNQNXXXXXXQ

[0136] The oligomeric structures shown in Tables 1 to 13 are double stranded structures, composed of a first strand (top) and a second strand (bottom). Each of the first and second strands is an oligomeric molecule that, by itself, can be a single stranded molecule. The single stranded molecules can be active for modulating gene expression.

[0137] An oligomeric compound of this invention may have any one of the structures shown in Tables 1 to 13. An oligomeric compound of this invention may have any one of the structures shown in Tables 1 to 13, where the structure has a nucleobase sequence targeted to a corresponding nucleic acid target.

[0138] An oligomeric compound of this invention may have any one of the sequences shown in Tables 1 to 13.

[0139] An oligomeric compound of this invention may be any one of the strands shown in Tables 1 to 13.

[0140] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer.

[0141] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is less than twenty.

[0142] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is less than twelve.

[0143] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is less than ten.

[0144] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is less than eight.

[0145] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is from 1 to 20.

[0146] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is from 1 to 15.

[0147] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is from 1 to 9.

[0148] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide.

[0149] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is less than twenty.

[0150] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is less than twelve.

[0151] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is less than ten.

[0152] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is less than eight.

[0153] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is from 1 to 20.

[0154] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is from 1 to 15.

[0155] In some embodiments, an oligomeric compound of this invention may have a first strand and a second strand, each of the strands independently being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is from 1 to 9.

[0156] An oligomeric compound of this invention may be a single stranded molecule, wherein the single strand is any one of the strands shown in Tables 1 to 13.

[0157] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer.

[0158] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is less than twenty.

[0159] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is less than twelve.

[0160] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is less than ten.

[0161] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is less than eight.

[0162] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is from 1 to 20.

[0163] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is from 1 to 15.

[0164] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a Q monomer, and where the number of Q monomers is from 1 to 9.

[0165] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide.

[0166] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is less than twenty.

[0167] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is less than twelve.

[0168] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is less than ten.

[0169] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is less than eight.

[0170] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is from 1 to 20.

[0171] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is from 1 to 15.

[0172] In some embodiments, an oligomeric compound of this invention may have a single strand being 19-23 monomers in length, where any monomer that is not a UNA monomer can be a 2-O-Methyl modified ribonucleotide, and where the number of 2-O-Methyl modified ribonucleotides is from 1 to 9.

[0173] Methods for Treating Disease

[0174] Methods of this invention include the treatment and prevention of various diseases in mammalian subjects. A subject can be human.

[0175] In the methods of this invention, a subject in need of treatment or prevention can be administered an effective amount of an oligomeric compound of this invention. A subject can be a human or mammal.

[0176] An effective amount of an oligomeric compound of this invention can be a dose ranging from 0.001 mg/kg to 50.0 mg/kg.

[0177] In the methods of this invention, target mRNA expression can be reduced in a subject for at least 5 days. In certain embodiments, target mRNA expression can be reduced in a subject for at least 10 days, or 15 days.

[0178] In the methods of this disclosure, the administration of an oligomeric compound of this invention may not result in an inflammatory response.

[0179] In further embodiments, this invention includes methods for inhibiting expression of a target gene in a cell, by treating the cell with an oligomeric compound of this invention.

[0180] In additional embodiments, this invention includes methods for inhibiting expression of a target gene in a mammal, by administering to the mammal a composition containing a UNA oligomer.

[0181] UNA Oligomers for TTR

[0182] Embodiments of this invention can provide oligomeric molecules that are active agents targeted to TTR.

[0183] Examples of UNA oligomers of this invention that are targeted to TTR are shown in Tables 14 to 17. Tables 14 and 15 represent sense and antisense pairs, and Tables 16 and 17 represent additional sense and antisense pairs. For example, SEQ ID Nos:237 and 238 are a sense and antisense pair.

TABLE-US-00015 TABLE14 UNAoligomerstargetedtoTTR SEQ ID NO Strand UNAoligomers(5 -3) 237 Sense mGrUmArAmCrCmArArGrAmGrUmArUmUrCm CrAmU 238 Anti mUrGmGrAmArUmArCmUrCmUmUmGrGmUrUmAr CmAmU 239 Sense mGrUmArAmCrCmArArGrAmGrUmArUmUrCm CrAmU 240 Anti mUrGmGrAmArUmArCmUrCmUmUmGrGmUrUmAr CmAmU 241 Sense *mG*rUmArAmCrCmArArGrAmGrUmArUmUr CmCA**mU 242 Anti mU*G*mGrAmArUmArCmUrCmUmUmGrGmUrUm ArCmA**mU 243 Sense *mG*U*mArAmCrCmArArGrAmGrUmArUmUr CmC*A**mU 244 Anti mU*G*mG*rAmArUmArCmUrCmUmUmGrGmUrUm AC*mA**mU 245 Sense *mG*2FU*mA2FAmC2FCmA2FA2FG2FAmG2F UmA2FUmU2FCmC*2FA**2FU 246 Anti mU*2FG*mG*2FAmA2FUmA2FCmU2FCmUmUmG2 FGmU2FUmA2FC*mA**mU 247 Sense *mG*U*mArAmCrCmArArGrAmGrUmArUmUr CmC*A**mU 248 Anti mU*G*mG*rAmArUmArCmUrCmUmUmGrGmUrUm AC*mU**mU

TABLE-US-00016 TABLE15 UNAoligomerstargetedtoTTR SEQ ID NO Strand UNAoligomers(5 -3) 249 Sense mGrUmArAmCrCArArGrAGrUArUUrCmCrA mU 250 Anti mUrGmGrAArUArCUrCUmUmGrGmUrUmArCCrA mU 251 Sense mGrUArACrCmArArGrAGrUArUmUrCCrAmU 252 Anti mUrGmGrAArUArCmUrCUmUmGrGUrUArCmA mU 253 Sense *G*rUmArAmCrCArArGrAGrUArUUrCCA** mU 254 Anti mU*G*mGrAmArUArCUrCUUGrGUrUmArCmA* *mU 255 Sense *mG*U*mArACrCArArGrAGrUArUUrCC*A* *mU 256 Anti mU*G*mG*rAmArUArCUrCUUGrGUrUAC*mA* *mU 257 Sense *mG*2FU*mA2FAmC2FCmA2FA2FG2FAmG2FU mA2FUmU2FCmC*2FA**2FU 258 Anti mU*2FG*mG*2FAmA2FUmA2FCmU2FCmUmUmG2 FGmU2FUmA2FC*mA**mU 259 Sense *mG*U*mArACrCArArGrAGrUArUUrCmC*A* *mU 260 Anti mU*G*mG*rAArUArCUrCUUGrGUrUmAC*mU* *mU

TABLE-US-00017 TABLE16 UNAoligomerstargetedtoTTR SEQ ID NO Strand UNAoligomers(5 -3) 261 Sense mArAmCrCmArAmGrArGrUmArUmUrCmCrAm UrUmU 262 Anti mArAmUrGmGrAmArUmArCmUmCmUrUmGrGmUr UmCmU 263 Sense mArAmCrCmArAmGrArGrUmArUmUrCmCrAm UrUmU 264 Anti mArAmUrGmGrAmArUmArCmUmCmUrUmGrGmUr UmCmU 265 Sense *mA*rAmCrCmArAmGrArGrUmArUmUrCmCr AmUU**mU 266 Anti mA*A*mUrGmGrAmArUmArCmUmCmUrUmGrGm UrUmC**mU 267 Sense *mA*A*mCrCmArAmGrArGrUmArUmUrCmCr AmU*U**mU 268 Anti mA*A*mU*rGmGrAmArUmArCmUmCmUrUmGrGm UU*mC**mU 269 Sense *mA*2FA*mC2FCmA2FAmG2FA2FG2FUmA2FU mU2FCmC2FAmU*2FU**2FU 270 Anti mA*2FA*mU*2FGmG2FAmA2FUmA2FCmUmCmU2 FUmG2FGmU2FU*mC**mU 271 Sense C*mA*A*mCrCmArAmGrArGrUmArUmUrCmCr AmU*U**mU 272 Anti mA*A*mU*rGmGrAmArUmArCmUmCmUrUmGr GmUU*mG**mU

TABLE-US-00018 TABLE17 UNAoligomerstargetedtoTTR SEQ ID NO Strand UNAoligomers(5 -3) 273 Sense mArAmCrCmArAGrArGrUArUUrCCrAmUrUm U 274 Anti mArAmUrGmGrAmArUArCUCUrUmGrGmUrUmC mU 275 Sense mArAmCrCArAGrArGrUArUUrCCrAmUrUmU 276 Anti mArAmUrGmGrAArUArCUCUrUmGrGmUrUmCm U 277 Sense *mA*rAmCrCArAGrArGrUArUUrCCrAUU** mU 278 Anti mA*A*mUrGmGrAArUArCUCUrUGrGmUrUmC* *mU 279 Sense *mA*A*mCrCArAGrArGrUArUUrCCrAU*U* *mU 280 Anti mA*A*mU*rGmGrAArUArCUCUrUGrGUU*mC* *mU 281 Sense *mA*2FA*mC2FCmA2FAmG2FA2FG2FUmA2FU mU2FCmC2FAmU*2FU**2FU 282 Anti mA*2FA*mU*2FGmG2FAmA2FUmA2FCmUmCmU2 FUmG2FGmU2FU*mC**mU 283 Sense C*mA*A*mCrCArAGrArGrUArUUrCCrAmU*U* *mU 284 Anti mA*A*mU*rGGrAArUArCUmCmUrUGrGUU*G* *mU

[0184] For example, a UNA oligomer may have a strand being SEQ ID NO:87. A UNA oligomer may have a strand being SEQ ID NO:88. A UNA oligomer may have a strand being SEQ ID NO:87, and a strand being SEQ ID NO:88. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of TTR. These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:237 to 284.

[0185] For example, a UNA oligomer may have a strand being SEQ ID NO:105. A UNA oligomer may have a strand being SEQ ID NO:106. A UNA oligomer may have a strand being SEQ ID NO:105, and a strand being SEQ ID NO:106. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of TTR. These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:237 to 284.

[0186] A UNA oligomer may comprise a first strand being SEQ ID NO:239 and a second strand being SEQ ID NO:240.

[0187] A UNA oligomer may comprise a first strand being SEQ ID NO:263 and a second strand being SEQ ID NO:264.

[0188] UNA Oligomers for APOB

[0189] Embodiments of this invention can provide oligomeric molecules that are active agents targeted to APOB.

[0190] Examples of UNA oligomers of this invention that are targeted to TTR are shown in Tables 18 and 19. Tables 18 and 19 represent sense and antisense pairs. For example, SEQ ID NOs:285 and 286 are a sense and antisense pair.

TABLE-US-00019 TABLE18 UNAoligomerstargetedtoAPOB SEQ ID NO Strand UNAoligomers(5 -3) 285 Sense rGrGrArArUrCrUrUrArUrArUrUrUrGrArUr CrCrArA 286 Anti rUrUrGrGrArUrCrArArArUrArUrArArGrAr UrUrCrC 287 Sense mGrAmArUmCrUmUrArUrAmUrUmUrGmArUm CrCmArAmU 288 Anti mUrUmGrGmArUmCrAmArAmUmAmUrAmArGmAr UmUrCmCmU 289 Sense rGrGrArArUrCmUmUrAmUrAmUmUmUrGrArUr CmCrArAmU 290 Anti mUmUrGrGrArUmCrArArAmUrAmUrArArGrAm UrUrCmCmU 291 Sense rGrGrArArUrCmUmUrAmUrAmUmUmUrGrArUr CmCrArAmU 292 Anti mUrUmGrGmArUmCrAmArAmUmAmUrAmArGmAr UmUrCmCmU 293 Sense mGrAmArUmCrUmUrArUrAmUrUmUrGmArUm CrCmArAmU 294 Anti mUmUrGrGrArUmCrArArAmUrAmUrArArGrAm UrUrCmCmU

TABLE-US-00020 TABLE19 UNAoligomerstargetedtoAPOB SEQ ID NO Strand UNAoligomers(5 -3) 295 Sense rGrGrArArUrCrUrUrArUrArUrUrUrGrAr UrCrCrArA 296 Anti rUrUrGrGrArUrCrArArArUrArUrArArGr ArUrUrCrC 297 Sense mGrAmArUmCrUUrArUrAUrUUrGArUCrCm ArAmU 298 Anti mUrUmGrGmArUmCrAArAUAUrAArGmArUmUr CmCmU 299 Sense rGrGrArArUrCmUmUrAmUrAUUUrGrArUrCC rArAmU 300 Anti mUmUrGrGrArUmCrArArAUrAmUrArArGrAm UrUrCmCmU 301 Sense rGrGrArArUrCmUmUrAUrAUUUrGrArUrCCr ArAmU 302 Anti mUrUmGrGmArUCrAArAUAUrAArGArUmUrCm CmU 303 Sense mGrAmArUCrUUrArUrAUrUUrGArUCrCArA mU 304 Anti mUmUrGrGrArUCrArArAUrAUrArArGrAmUr UrCmCmU

[0191] For example, a UNA oligomer may have a strand being SEQ ID NO:87. A UNA oligomer may have a strand being SEQ ID NO:88. A UNA oligomer may have a strand being SEQ ID NO:87, and a strand being SEQ ID NO:88. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of APOB. These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:285 to 304.

[0192] For example, a UNA oligomer may have a strand being SEQ ID NO:105. A UNA oligomer may have a strand being SEQ ID NO:106. A UNA oligomer may have a strand being SEQ ID NO:105, and a strand being SEQ ID NO:106. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of APOB. These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:285 to 304.

[0193] A UNA oligomer may comprise a first strand being SEQ ID NO:287 and a second strand being SEQ ID NO:288.

[0194] A UNA oligomer may comprise a first strand being SEQ ID NO:291 and a second strand being SEQ ID NO:292.

[0195] UNA Oligomers for PCSK9.

[0196] Embodiments of this invention can provide oligomeric molecules that are active agents targeted to PCSK9.

[0197] Examples of UNA oligomers of this invention that are targeted to PCSK9 are shown in Tables 20 and 21. Tables 20 and 21 represent sense and antisense pairs. For example, SEQ ID NOs:305 and 306 are a sense and antisense pair.

TABLE-US-00021 TABLE20 UNAoligomerstargetedtoPCSK9 SEQ ID NO Strand UNAoligomers(5 -3) 305 Sense mCrCmUrGmGrAmGrUrUrUmArUmUrCmGrGm ArAmU 306 Anti mUrUmCrCmGrAmArUmArAmAmCmUrCmCrAmGr GmCmU 307 Sense mCmCmUrGrGrArGmUmUmUrAmUmUmCrGrGr ArAmU 308 Anti mUrUmCrCmGrAmArUmArAmAmCmUrCmCrAmGr GmCmU 309 Sense rGrCrCrUrGrGrArGrUrUrUrArUrUrCrGr GrArAmU 310 Anti rUrUrCrCrGrArArUrArArArCrUrCrCrArGr GrCmU

TABLE-US-00022 TABLE21 UNAoligomerstargetedtoPCSK9 SEQ ID NO Strand UNAoligomers(5 -3) 311 Sense mCrCmUrGmGrAGrUrUrUArUUrCGrGmArAm U 312 Anti mUrUmCrCmGrAmArUArAACUrCmCrAmGrGmC mU 313 Sense mCmCUrGrGrArGUUUrAUUmCrGrGrArAmU 314 Anti mUrUmCrCmGrAArUArAACUrCmCrAmGrGmCm U 315 Sense mGrCmCrUrGrGrArGrUrUrUrArUrUrCrGr GrArAmU 316 Anti rUmUrCrCrGrArAmUmAmAmArCrUrCrCrArGr GrCmU

[0198] For example, a UNA oligomer may have a strand being SEQ ID NO:87. A UNA oligomer may have a strand being SEQ ID NO:88. A UNA oligomer may have a strand being SEQ ID NO:87, and a strand being SEQ ID NO:88. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of PCSK9. These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:305 to 316.

[0199] For example, a UNA oligomer may have a strand being SEQ ID NO:105. A UNA oligomer may have a strand being SEQ ID NO:106. A UNA oligomer may have a strand being SEQ ID NO:105, and a strand being SEQ ID NO:106. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of PCSK9. These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:305 to 316.

[0200] A UNA oligomer may comprise a first strand being SEQ ID NO:305 and a second strand being SEQ ID NO:306.

[0201] UNA Oligomers for APOC3.

[0202] Embodiments of this invention can provide oligomeric molecules that are active agents targeted to APOC3.

[0203] Examples of UNA oligomers of this invention that are targeted to APOCIII are shown in Tables 22 and 23. Tables 22 and 23 represent sense and antisense pairs. For example, SEQ ID NOs:317 and 318 are a sense and antisense pair.

TABLE-US-00023 TABLE22 UNAoligomerstargetedtoAPOC3 SEQ ID NO Strand UNAoligomers(5 -3) 317 Sense rArArArGrGrGrArCrArGrUrArUrUrCrUr CrAmU 318 Anti rUrGrArGrArArUrArCrUrGrUrCrCrCrUr UrUrUmU 319 Sense *mArAmArGmGrGmArCrArGmUrAmUrUmCr UmCrA*mU 320 Anti mU*rGmArGmArAmUrAmCrUmGmUmCrCmCrUm UrUmU*mU 321 Sense mArAmArGmGrGmArCrArGmUrAmUrUmCrUm CrAmU 322 Anti mUrGmArGmArAmUrAmCrUrGrUrCrCmCrUm UrUmUmU 323 Sense *mArAmArGmGrGmArCrArGmUrAmUrUmCr UmCrA*mU 324 Anti mU*rGmArGmArAmUrAmCrUrGrUrCrCmCrUm UrUmU*mU

TABLE-US-00024 TABLE23 UNAoligomerstargetedtoAPOC3 SEQ ID NO Strand UNAoligomers(5 -3) 325 Sense rArArArGrGrGrArCrArGrUrArUrUrCrUr CrAmU 326 Anti rUrGrArGrArArUrArCrUrGrUrCrCrCrUrUr UrUmU 327 Sense *mArAmArGmGrGArCrArGUrAUrUCrUmCrA *mU 328 Anti mU*rGmArGmArAmUrACrUGUCrCmCrUmUrUmU *mU 329 Sense mArAmArGGrGArCrArGUrAUrUCrUmCrAmU 330 Anti mUrGmArGmArAmUrACrUrGrUrCrCCrUmUrUm UmU 331 Sense *mArAmArGGrGArCrArGUrAUrUCrUCrA*m U 332 Anti mU*rGmArGmArAUrACrUrGrUrCrCCrUmUrUm U*mU

[0204] For example, a UNA oligomer may have a strand being SEQ ID NO:87. A UNA oligomer may have a strand being SEQ ID NO:88. A UNA oligomer may have a strand being SEQ ID NO:87, and a strand being SEQ ID NO:88. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of APOC3. These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:317 to 332.

[0205] For example, a UNA oligomer may have a strand being SEQ ID NO:105. A UNA oligomer may have a strand being SEQ ID NO:106. A UNA oligomer may have a strand being SEQ ID NO:105, and a strand being SEQ ID NO:106. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of APOC3. These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:317 to 332.

[0206] A UNA oligomer may comprise a first strand being SEQ ID NO:319 and a second strand being SEQ ID NO:320.

[0207] A UNA oligomer may comprise a first strand being SEQ ID NO:321 and a second strand being SEQ ID NO:322.

[0208] UNA Oligomers for HBV

[0209] Embodiments of this invention can provide oligomeric molecules that are active agents targeted to HBV.

[0210] Examples of UNA oligomers of this invention that are targeted to HBV are shown in Tables 24 to 27. Tables 24 and 25 represent sense and antisense pairs, and Tables 26 and 27 represent additional sense and antisense pairs. For example, SEQ ID NOs:333 and 353 are a sense and antisense pair, and so on through SEQ ID NOs:352 and 372.

TABLE-US-00025 TABLE24 UNAoligomerstargetedtoHBV(Sense) REF SEQ POS IDNO Sense(5-3)SEQIDNOS:861to880 1525 333 UNA-C/mGCmACmCUmCUCUmUUmACmGCmGG/ UNA-U/mU 251 334 UNA-G/mACmUCmGUmGGUGmGAmCUmUCmUC/ UNA-U/mU 254 335 UNA-U/mCGmUGmGUmGGACmUUmCUmCUmCA/ UNA-U/mU 374 336 UNA-U/mGGmAUmGUmGUCUmGCmGGmCGmUU/ UNA-U/mU 1575 337 UNA-C/mCGmUGmUGmCACUmUCmGCmUUmCA/ UNA-U/mU 1577 338 UNA-G/mUGmUGmCAmCUUCmGCmUUmCAmCC/ UNA-U/mU 1578 339 UNA-U/mGUmGCmACmUUCGmCUmUCmACmCU/ UNA-U/mU 1579 340 UNA-G/mUGmCAmCUmUCGCmUUmCAmCCmUC/ UNA-U/mU 1581 341 UNA-G/mCAmCUmUCmGCUUmCAmCCmUCmUG/ UNA-U/mU 247 342 UNA-U/mCUmAGmACmUCGUmGGmUGmGAmCU/ UNA-U/mU 248 343 UNA-C/mUAmGAmCUmCGUGmGUmGGmACmUU/ UNA-U/mU 249 344 UNA-U/mAGmACmUCmGUGGmUGmGAmCUmUC/ UNA-U/mU 250 345 UNA-A/mGAmCUmCGmUGGUmGGmACmUUmCU/ UNA-U/mU 1776 346 UNA-G/mGAmGGmCUmGUAGmGCmAUmAAmAU/ UNA-U/mU 1777 347 UNA-G/mAGmGCmUGmUAGGmCAmUAmAAmUU/ UNA-U/mU 1779 348 UNA-G/mGCmUGmUAmGGCAmUAmAAmUUmGG/ UNA-U/mU 1780 349 UNA-G/mCUmGUmAGmGCAUmAAmAUmUGmGU/ UNA-U/mU 1781 350 UNA-C/mUGmUAmGGmCAUAmAAmUUmGGmUC/ UNA-U/mU 1782 351 UNA-U/mGUmAGmGCmAUAAmAUmUGmGUmCU/ UNA-U/mU 256 352 UNA-G/mUGmGUmGGmACUUmCUmCUmCAmAU/ UNA-U/mU

TABLE-US-00026 TABLE25 UNAoligomerstargetedtoHBV(Antisense) REF SEQ Antisense(5-3)SEQIDNOS: POS IDNO 881to900 1525 353 mCCmGCmGUmAAmAGmAmGmAGmGUmGCmG/ UNA-U/mU 251 354 mGAmGAmAGmUCmCAmCmCmACmGAmGUmC/ UNA-U/mU 254 355 mUGmAGmAGmAAmGUmCmCmACmCAmCGmA/ UNA-U/mU 374 356 mAAmCGmCCmGCmAGmAmCmACmAUmCCmA/ UNA-U/mU 1575 357 mUGmAAmGCmGAmAGmUmGmCAmCAmCGmG/ UNA-U/mU 1577 358 mGGmUGmAAmGCmGAmAmGmUGmCAmCAmC/ UNA-U/mU 1578 359 mAGmGUmGAmAGmCGmAmAmGUmGCmACmA/ UNA-U/mU 1579 360 mGAmGGmUGmAAmGCmGmAmAGmUGmCAmC/ UNA-U/mU 1581 361 mCAmGAmGGmUGmAAmGmCmGAmAGmUGmC/ UNA-U/mU 247 362 mAGmUCmCAmCCmACmGmAmGUmCUmAGmA/ UNA-U/mU 248 363 mAAmGUmCCmACmCAmCmGmAGmUCmUAmG/ UNA-U/mU 249 364 mGAmAGmUCmCAmCCmAmCmGAmGUmCUmA/ UNA-U/mU 250 365 mAGmAAmGUmCCmACmCmAmCGmAGmUCmU/ UNA-U/mU 1776 366 mAUmUUmAUmGCmCUmAmCmAGmCCmUCmC/ UNA-U/mU 1777 367 mAAmUUmUAmUGmCCmUmAmCAmGCmCUmC/ UNA-U/mU 1779 368 mCCmAAmUUmUAmUGmCmCmUAmCAmGCmC/ UNA-U/mU 1780 369 mACmCAmAUmUUmAUmGmCmCUmACmAGmC/ UNA-U/mU 1781 370 mGAmCCmAAmUUmUAmUmGmCCmUAmCAmG/ UNA-U/mU 1782 371 mAGmACmCAmAUmUUmAmUmGCmCUmACmA/ UNA-U/mU 256 372 mAUmUGmAGmAGmAAmGmUmCCmACmCAmC/ UNA-U/mU

TABLE-US-00027 TABLE26 UNAoligomerstargetedtoHBV(Sense) REF SEQ POS IDNO Sense(5-3)SEQIDNOS:901to921 1863 373 UNA-U/mUCmAAmGCmCUCCmAAmGCmUGmUG/ UNA-U/mU 1864 374 UNA-U/mCAmAGmCCmUCCAmAGmCUmGUmGC/ UNA-U/mU 1865 375 UNA-C/mAAmGCmCUmCCAAmGCmUGmUGmCC/ UNA-U/mU 1866 376 UNA-A/mAGmCCmUCmCAAGmCUmGUmGCmCU/ UNA-U/mU 376 377 UNA-G/mAUmGUmGUmCUGCmGGmCGmUUmUU/ UNA-U/mU 378 378 UNA-U/mGUmGUmCUmGCGGmCGmUUmUUmAU/ UNA-U/mU 380 379 UNA-U/mGUmCUmGCmGGCGmUUmUUmAUmCA/ UNA-U/mU 1818 380 UNA-A/mACmUUmUUmUCACmCUmCUmGCmCU/ UNA-U/mU 244 381 UNA-G/mAGmUCmUAmGACUmCGmUGmGUmGG/ UNA-U/mU 245 382 UNA-A/mGUmCUmAGmACUCmGUmGGmUGmGA/ UNA-U/mU 246 383 UNA-G/mUCmUAmGAmCUCGmUGmGUmGGmAC/ UNA-U/mU 409 384 UNA-C/mAUmCCmUGmCUGCmUAmUGmCCmUC/ UNA-U/mU 411 385 UNA-U/mCCmUGmCUmGCUAmUGmCCmUCmAU/ UNA-U/mU 412 386 UNA-C/mCUmGCmUGmCUAUmGCmCUmCAmUC/ UNA-U/mU 413 387 UNA-C/mUGmCUmGCmUAUGmCCmUCmAUmCU/ UNA-U/mU 414 388 UNA-U/mGCmUGmCUmAUGCmCUmCAmUCmUU/ UNA-U/mU 252 389 UNA-A/mCUmCGmUGmGUGGmACmUUmCUmCU/ UNA-U/mU 253 390 UNA-C/mUCmGUmGGmUGGAmCUmUCmUCmUC/ UNA-U/mU 1576 391 UNA-C/mGUmGUmGCmACUUmCGmCUmUCmAC/ UNA-U/mU 1580 392 UNA-U/mGCmACmUUmCGCUmUCmACmCUmCU/ UNA-U/mU 1582 393 UNA-C/mACmUUmCGmCUUCmACmCUmCUmGC/ UNA-U/mU

TABLE-US-00028 TABLE27 UNAoligomerstargetedtoHBV(Antisense) REF SEQ Antisense(5-3)SEQIDNOS: POS IDNO 922to942 1863 394 mCAmCAmGCmUUmGGmAmGmGCmUUmGAmA/ UNA-U/mU 1864 395 mGCmACmAGmCUmUGmGmAmGGmCUmUGmA/ UNA-U/mU 1865 396 mGGmCAmCAmGCmUUmGmGmAGmGCmUUmG/ UNA-U/mU 1866 397 mAGmGCmACmAGmCUmUmGmGAmGGmCUmU/ UNA-U/mU 376 398 mAAmAAmCGmCCmGCmAmGmACmACmAUmC/ UNA-U/mU 378 399 mAUmAAmAAmCGmCCmGmCmAGmACmACmA/ UNA-U/mU 380 400 mUGmAUmAAmAAmCGmCmCmGCmAGmACmA/ UNA-U/mU 1818 401 mAGmGCmAGmAGmGUmGmAmAAmAAmGUmU/ UNA-U/mU 244 402 mCCmACmCAmCGmAGmUmCmUAmGAmCUmC/ UNA-U/mU 245 403 mUCmCAmCCmACmGAmGmUmCUmAGmACmU/ UNA-U/mU 246 404 mGUmCCmACmCAmCGmAmGmUCmUAmGAmC/ UNA-U/mU 409 405 mGAmGGmCAmUAmGCmAmGmCAmGGmAUmG/ UNA-U/mU 411 406 mAUmGAmGGmCAmUAmGmCmAGmCAmGGmA/ UNA-U/mU 412 407 mGAmUGmAGmGCmAUmAmGmCAmGCmAGmG/ UNA-U/mU 413 408 mAGmAUmGAmGGmCAmUmAmGCmAGmCAmG/ UNA-U/mU 414 409 mAAmGAmUGmAGmGCmAmUmAGmCAmGCmA/ UNA-U/mU 252 410 mAGmAGmAAmGUmCCmAmCmCAmCGmAGmU/ UNA-U/mU 253 411 mGAmGAmGAmAGmUCmCmAmCCmACmGAmG/ UNA-U/mU 1576 412 mGUmGAmAGmCGmAAmGmUmGCmACmACmG/ UNA-U/mU 1580 413 mAGmAGmGUmGAmAGmCmGmAAmGUmGCmA/ UNA-U/mU 1582 414 mGCmAGmAGmGUmGAmAmGmCGmAAmGUmG/ UNA-U/mU

[0211] For example, a UNA oligomer may have a strand being SEQ ID NO:87. A UNA oligomer may have a strand being SEQ ID NO:88. A UNA oligomer may have a strand being SEQ ID NO:87, and a strand being SEQ ID NO:88. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of a viral gene of Hepatitis B virus (HBV). These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs:333 to 414.

[0212] For example, a UNA oligomer may have a strand being SEQ ID NO:105. A UNA oligomer may have a strand being SEQ ID NO:106. A UNA oligomer may have a strand being SEQ ID NO:105, and a strand being SEQ ID NO:106. These UNA oligomers can have a nucleobase sequence corresponding to a target sequence of a viral gene of Hepatitis B virus (HBV). These UNA oligomers can have a nucleobase sequence corresponding to any of SEQ ID NOs: 333 to 414.

[0213] Methods for Treating Disease

[0214] Methods of this invention include the treatment and prevention of various diseases in mammalian subjects. A subject can be a human or mammal.

[0215] In the methods of this invention, a subject in need of treatment or prevention can be administered an effective amount of an oligomeric compound of this invention.

[0216] In addition, the UNA oligomers of this invention may provide increased activity in vitro, as well as increased potency in vivo.

[0217] Further, the UNA oligomers of this invention may provide increased enzymatic stability.

[0218] Moreover, the UNA oligomers of this invention can provide long lasting activity in vitro, as well as long lasting potency in vivo.

[0219] A UNA oligomer of this invention can retain at least 50% activity in vitro six days after transfection.

[0220] A UNA oligomer of this invention can retain at least 80% activity in vitro six days after transfection.

[0221] A UNA oligomer of this invention can retain at least 30% potency in vivo six days after administration.

[0222] A UNA oligomer of this invention can retain at least 50% potency in vivo six days after administration.

[0223] UNA oligomers of this invention can provide long acting properties, and reduce the dose levels required for efficacious therapy.

[0224] An effective amount of an oligomeric compound of this invention can be a dose ranging from 0.001 mg/kg to 50.0 mg/kg.

[0225] In the methods of this invention, target mRNA expression can be reduced in a subject for at least 5 days. In certain embodiments, target mRNA expression can be reduced in a subject for at least 10 days, or 15 days.

[0226] In the methods of this disclosure, the administration of an oligomeric compound may not result in an inflammatory response.

[0227] In further embodiments, this invention includes methods for inhibiting expression of a target gene in a cell, by treating the cell with an oligomeric compound of this invention.

[0228] In additional embodiments, this invention includes methods for inhibiting expression of a target gene in a mammal, by administering to the mammal a composition containing an oligomeric compound of this invention.

[0229] In some embodiments, a UNA oligomer targeted to a transthyretin nucleic acid, TTR, can be used as an active agent for preventing or treating amyloid neuropathy, amyloidosis, or amyloidosis related to transthyretin in a subject in need thereof.

[0230] In further embodiments, a UNA oligomer targeted to an apolipoprotein B nucleic acid, APOB, can be used as an active agent for preventing or treating hypercholesterolemia, or cholesterol disorder in a subject in need thereof.

[0231] In additional embodiments, a UNA oligomer targeted to a proprotein convertase subtilisin/kexin type 9 nucleic acid, PCSK9, can be used as an active agent for preventing or treating hypercholesterolemia, or cholesterol disorder in a subject in need thereof.

[0232] In certain embodiments, a UNA oligomer targeted to an apolipoprotein C-III nucleic acid, APOC3 or APOCIII, can be used as an active agent for preventing or treating hypertriglyceridemia, or lipoprotein disorder in a subject in need thereof.

[0233] In some embodiments, a UNA oligomer targeted to a sequence of an HBV genome, HBV (Hepatitis B virus), can be used as an active agent for preventing or treating a disease associated with HBV infection, in a subject in need thereof.

[0234] Pharmaceutical Compositions

[0235] In some aspects, this invention provides pharmaceutical compositions containing an oligomeric compound and a pharmaceutically acceptable carrier.

[0236] A pharmaceutical composition can be capable of local or systemic administration. In some aspects, a pharmaceutical composition can be capable of any modality of administration. In certain aspects, the administration can be intravenous, subcutaneous, pulmonary, intramuscular, intraperitoneal, dermal, oral, or nasal administration.

[0237] Embodiments of this invention include pharmaceutical compositions containing an oligomeric compound in a lipid formulation.

[0238] In some embodiments, a pharmaceutical composition may comprise one or more lipids selected from cationic lipids, anionic lipids, sterols, pegylated lipids, and any combination of the foregoing.

[0239] In certain embodiments, a pharmaceutical composition can be substantially free of liposomes.

[0240] In further embodiments, a pharmaceutical composition can include liposomes or nanoparticles.

[0241] Some examples of lipids and lipid compositions for delivery of an active molecule of this invention are given in WO/2015/074085, which is hereby incorporated by reference in its entirety.

[0242] In additional embodiments, a pharmaceutical composition can contain an oligomeric compound within a viral or bacterial vector.

[0243] A pharmaceutical composition of this disclosure may include carriers, diluents or excipients as are known in the art. Examples of pharmaceutical compositions are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro ed. 1985).

[0244] Examples of excipients for a pharmaceutical composition include antioxidants, suspending agents, dispersing agents, preservatives, buffering agents, tonicity agents, and surfactants.

EXAMPLES

Example 1: Luciferase Reporter Assay

[0245] Luciferase-based reporter plasmid was constructed based on psiCHECK2 vector (Promega, Madison, Wis.). Reporter p(1-20) was generated with oligonucleotides containing the sequence from position 1 through 2500 relative to Eco RI digestion site cloned into the multiple cloning region downstream of the stop codon of the SV40 promoted Renilla luciferase gene in psiCHECK2, which made the expression of Renilla luciferase gene under the regulation of the artificial 3UTR sequence. Renilla luciferase activity was then used as an indicator of the effect of the artificial 3UTR on transcript stability and translation efficiency. The psiCHECK-2 Vector also contained a constitutively expressed Firefly luciferase gene, which served as an internal control to normalize transfection efficiency.

[0246] A total of 5,000 HepB3 cells (American Type Culture Collection) were plated onto a well of 96-well plate one day before the transfection. The cells were incubated at 37 C. in 100 l of DMEM (Life Technologies, Carlsbad, Calif.) supplemented with 0.1 mM nonessential amino acids and 10% FBS (Life Technologies, Carlsbad, Calif.). The culture medium was changed to 90 l of fresh medium just before the transfection. The reporter plasmid and UNA Oligomer were co-transfected with transfection reagent, Lipofectamine 3000 (Life Technologies, Carlsbad, Calif.) was used to transfect reporter plasmid (100 ng) and a various amount of UNA Oligomer together with P3000 into the cells according to manufacturer's instruction.

[0247] Dual-Luciferase Reporter Assay System (DLR assay system, Promega, Madison, Wis.) was used to perform dual-reporter assays on psiCHECK2 based reporter systems. Twenty-four hours after transfection, the cells were washed gently with phosphate buffered saline once. A 50 l well of Passive Lysis Buffer (Promega, Madison, Wis.) was added to the cells and incubated with gentle rocking for 20 min at room temperature. Luciferase activities were measured using Cytation 3 imaging reader (BioTek, Winooski, Vt.) and the effect of the UNA Oligomer on reporter expression was calculated based on ratio of Renilla/Firefly to normalize cell number and transfection efficiency.

Example 2: UNA Oligomer Stability

[0248] Protocol for determining the stability of a UNA oligomer measured against snake venom PDE I or FBS. The UNA oligomer (1-10 uM) is incubated with 210.sup.4 units/L Phosphodiesterase I from Crotalus adamanteus venom (code#: VPH, Worthington Biochemial, USA) in 20 mM Tris-HCl (pH 8.0), 100 mM NaCl, 15 mM MgCl2 or 10-50% of FBS (final concentration) at 37 C. After 0.5, 1 and 2 h, aliquots (3 L) are taken from the mixture and the reaction is stopped by mixing with 5.3 L of 0.5 M EDTA (pH 8.0). The sample is loaded on a 15% native polyacrylamide gel for electrophoresis. The gel is stained with SYBR SAFE and visualized under UV.

Example 3: UNA Oligomer Longevity

[0249] Protocol for determining the longevity of a UNA oligomer in vitro or in vivo. Total RNA samples from cell lines or mouse tissue are isolated. Reverse transcription reactions are performed using SuperScript III First-Strand (Life Technologies) with RNA samples, 50 nM stem-loop RT primer. All Reverse transcriptase reactions are run in triplicate. Real-time PCR is performed using a standard TaqMan PCR kit protocol with 400 nM universal primer, 400 nM gene-specific primer, and 300 nM TaqMan probe on an Applied Biosystems 7900HT Sequence Detection System. One l of cDNA template is added to a final volume of 25 l reaction. All reactions are carried out in triplicate with no template control as well as no-RT sample. The reactions are incubated in a 384-well plate at 95 C. for 10 min, followed by 40 cycles of 95 C. for 15 s and 60 C. for 1 min. All reactions are run in triplicate. The threshold cycle (CT) value, which is defined as the fractional cycle number at which the fluorescence passes the fixed threshold, is converted into an absolute copy number using a standard curve from a synthetic UNA oligomer.

Example 4

[0250] A Dose dependent inhibitory effect of UNA oligomers targeted to HBV was observed in Hep3B cells transfected with a reporter construct having HBV binding sites.

[0251] UNA oligomers of this invention were found to exhibit IC50 as shown in Table 28. In Table 28, the UNA oligomers correspond to the structures shown in Tables 24-27 having the same reference position.

TABLE-US-00029 TABLE 28 IC50 of UNA oligomers targeted to HBV IC50 pM Reference Hep3B cells No. Position (6 days) 1 244 917 2 245 328 3 246 816 4 247 5 248 148 6 249 7 250 8 251 554 9 252 374 10 253 703 11 254 44 12 256 8 13 374 1601 14 376 16 15 378 114 16 380 7 17 409 328 18 411 58 19 412 298 20 413 123 21 414 363 22 1525 23 1575 65 24 1576 137 25 1577 472 26 1578 63 27 1579 28 1580 255 29 1581 22 30 1776 461 31 1777 26 32 1779 348 33 1780 151 34 1781 227 35 1782 177 36 1818 49

[0252] The experimental results in Table 28 show that the UNA oligomers provide stable, surprisingly long-lasting activity for modulating gene expression. The activities of the UNA oligomers as measured by Luciferase reporter were in the picomolar range, six days after transfection.

Example 5

[0253] In addition, in experimental results the UNA oligomers of this invention targeted to HBV provide increased activity in vitro, as well as increased potency in vivo.

[0254] Further, in experimental results the UNA oligomers of this invention targeted to HBV provide increased enzymatic stability.

[0255] Moreover, in experimental results the UNA oligomers of this invention targeted to HBV provide long lasting activity in vitro, as well as long lasting potency in vivo.

[0256] UNA oligomers of this invention targeted to HBV provide long acting properties, and reduce the dose levels required for efficacious therapy.

Example 6

[0257] A dose dependent inhibitory effect of UNA oligomers targeted to TTR was observed in HepG2 cells, as shown in Table 29

TABLE-US-00030 TABLE 29 IC50 of UNA oligomers targeted to TTR IC50 pM (TTR) Structure HepG2 cells SEQ ID NOs: 237 and 238 5.23 SEQ ID NOs: 239 and 240 3.89 SEQ ID NOs: 241 and 242 4.98 SEQ ID NOs: 243 and 244 7.20 SEQ ID NOs: 245 and 246 24.8 SEQ ID NOs: 247 and 248 19.3

[0258] The experimental results in Table 29 show that TTR UNA oligomers provided stable, potent knockdown of TTR gene expression.

Example 7

[0259] A dose dependent inhibitory effect of TTR UNA oligomers was observed in HepG2 cells, as shown in Table 30.

TABLE-US-00031 TABLE 30 IC50 of UNA oligomers targeted to TTR IC50 pM (TTR) Structure HepG2 cells SEQ ID NOs: 261 and 262 3.20 SEQ ID NOs: 263 and 264 2.23 SEQ ID NOs: 265 and 266 3.36 SEQ ID NOs: 267 and 268 6.41 SEQ ID NOs: 269 and 270 6.67 SEQ ID NOs: 271 and 272 6.58

[0260] The experimental results in Table 30 show that TTR UNA oligomers provided stable, potent knockdown of TTR gene expression.

Example 8

[0261] In addition, in experimental results the UNA oligomers of this invention targeted to TTR provide increased activity in vitro, as well as increased potency in vivo.

[0262] Further, in experimental results the UNA oligomers of this invention targeted to TTR provide increased enzymatic stability.

[0263] Moreover, in experimental results the UNA oligomers of this invention targeted to TTR provide long lasting activity in vitro, as well as long lasting potency in vivo.

[0264] UNA oligomers of this invention targeted to TTR provide long acting properties, and reduce the dose levels required for efficacious therapy.

Example 9

[0265] A dose dependent inhibitory effect of UNA oligomers targeted to APOB was observed in mouse Hepa1-6 cells, as shown in Table 31.

TABLE-US-00032 TABLE 31 IC50 of UNA oligomers targeted to APOB IC50 nM (APOB) Structure Hepa1-6 cells SEQ ID NOs: 285 and 286 1.31 SEQ ID NOs: 287 and 288 0.48 SEQ ID NOs: 289 and 290 34 SEQ ID NOs: 291 and 292 0.68 SEQ ID NOs: 293 and 294 15

[0266] The experimental results in Table 31 show that APOB UNA oligomers provided stable, potent knockdown of APOB gene expression.

Example 10

[0267] A dose dependent inhibitory effect of UNA oligomers targeted to APOB was observed in mouse Hep3B cells, as shown in Table 32.

TABLE-US-00033 TABLE 32 IC50 of UNA oligomers targeted to APOB IC50 nM (APOB) Structure Hep3B cells SEQ ID NOs: 285 and 286 0.26 SEQ ID NOs: 287 and 288 0.80 SEQ ID NOs: 289 and 290 0.54 SEQ ID NOs: 291 and 292 0.21 SEQ ID NOs: 293 and 294 0.88

[0268] The experimental results in Table 32 show that APOB UNA oligomers provided stable, potent knockdown of APOB gene expression.

Example 11

[0269] In addition, in experimental results the UNA oligomers of this invention targeted to APOB provide increased activity in vitro, as well as increased potency in vivo.

[0270] Further, in experimental results the UNA oligomers of this invention targeted to APOB provide increased enzymatic stability.

[0271] Moreover, in experimental results the UNA oligomers of this invention targeted to APOB provide long lasting activity in vitro, as well as long lasting potency in vivo.

[0272] UNA oligomers of this invention targeted to APOB provide long acting properties, and reduce the dose levels required for efficacious therapy.

Example 12

[0273] A dose dependent inhibitory effect of UNA oligomers targeted to PCSK9 was observed in mouse Hepa1-6 cells, as shown in Table 33

TABLE-US-00034 TABLE 33 IC50 of UNA oligomers targeted to PCSK9 IC50 nM (PCSK9) Structure Hepa1-6 cells SEQ ID NOs: 305 and 306 2.37 SEQ ID NOs: 307 and 308 1.56 SEQ ID NOs: 309 and 310 1.78

[0274] The experimental results in Table 33 show that PCSK9 UNA oligomers provided stable, potent knockdown of PCSK9 gene expression.

Example 13

[0275] A dose dependent inhibitory effect of UNA oligomers targeted to PCSK9 was observed in mouse Hep3B cells, as shown in Table 34

TABLE-US-00035 TABLE 34 IC50 of UNA oligomers targeted to PCSK9 IC50 nM (PCSK9) Structure Hep3B cells SEQ ID NOs: 305 and 306 2.27 SEQ ID NOs: 307 and 308 0.70 SEQ ID NOs: 309 and 310 3.94

[0276] The experimental results in Table 34 show that PCSK9 UNA oligomers provided stable, potent knockdown of PCSK9 gene expression.

Example 14

[0277] In addition, in experimental results the UNA oligomers of this invention targeted to PCSK9 provide increased activity in vitro, as well as increased potency in vivo.

[0278] Further, in experimental results the UNA oligomers of this invention targeted to PCSK9 provide increased enzymatic stability.

[0279] Moreover, in experimental results the UNA oligomers of this invention targeted to PCSK9 provide long lasting activity in vitro, as well as long lasting potency in vivo.

[0280] UNA oligomers of this invention targeted to PCSK9 provide long acting properties, and reduce the dose levels required for efficacious therapy.

Example 15

[0281] A dose dependent inhibitory effect of UNA oligomers targeted to APOC3 was observed in mouse Hep3B cells, as shown in Table 35.

TABLE-US-00036 TABLE 35 IC50 of UNA oligomers targeted to APOC3 IC50 pM (APOC3) Structure Hep3B cells SEQ ID NOs: 317 and 318 2.8 SEQ ID NOs: 319 and 320 2.4 SEQ ID NOs: 321 and 322 3.4 SEQ ID NOs: 323 and 324 17.5

[0282] The experimental results in Table 35 show that APOC3 UNA oligomers provided stable, potent knockdown of APOC3 gene expression.

Example 16

[0283] In addition, in experimental results the UNA oligomers of this invention targeted to APOC3 provide increased activity in vitro, as well as increased potency in vivo.

[0284] Further, in experimental results the UNA oligomers of this invention targeted to APOC3 provide increased enzymatic stability.

[0285] Moreover, in experimental results the UNA oligomers of this invention targeted to APOC3 provide long lasting activity in vitro, as well as long lasting potency in vivo.

[0286] UNA oligomers of this invention targeted to APOC3 provide long acting properties, and reduce the dose levels required for efficacious therapy.

[0287] All publications, patents and literature specifically mentioned herein are incorporated by reference for all purposes.

[0288] It is understood that this invention is not limited to the particular methodology, protocols, materials, and reagents described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which will be encompassed by the appended claims.

[0289] It must be noted that as used herein and in the appended claims, the singular forms a, an, and the include plural reference unless the context clearly dictates otherwise. As well, the terms a (or an), one or more and at least one can be used interchangeably herein. It is also to be noted that the terms comprises, comprising, containing, including, and having can be used interchangeably.

[0290] Without further elaboration, it is believed that one skilled in the art can, based on the above description, utilize the present invention to its fullest extent. The following specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

[0291] All of the features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose.