SAA1-targeted oligomeric compounds, compositions and uses

Abstract

The technology relates in part to oligomeric compounds that can reduce SAA1 expression. The technology also relates in part to methods, and uses of the oligomeric compounds, for reducing SAA1 expression, and in certain instances treating a SAA1-associated medical condition.

Description

[0001] PATENT 7328*158807PCT / NLM-1005-PC SAA1-TARGETED OLIGOMERIC COMPOUNDS,

[0002] COMPOSITIONS AND USES RELATED APPLICATIONS

[0003] This Patent Convention Treaty (PCT) International Application claims the benefit of priority under 35 U. S. C. § 119(e) of U. S. Provisional Application Serial No. (USSN) 63 / 711,248, filed October 24, 2024; and USSN 63 / 768,693, filed March 7, 2025. The aforementioned application is expressly incorporated herein by reference in its entirety and for all purposes. All publications, patents, patent applications cited herein are hereby expressly incorporated by reference for all purposes.

[0004] TECHNICAL FIELD

[0005] The technology relates in part to oligomeric compounds that can reduce SAA1 expression. The technology also relates in part to methods, and uses of the oligomeric compounds, for reducing SAA1 expression, and in certain instances treating a SAA1-associated medical condition.

[0006] BACKGROUND

[0007] Less than 10% of rare diseases have approved treatments (Kaufmann et al., Orphanet J. Rare Dis. 13: 1-8 (2018)). Within rare diseases there are ultra-rare diseases with only 1-30 patients with the same pathogenic genetic variant worldwide (Crooke et al., Nat. Biotechnol. 39: 671-677 (2021)). The rarity of these diseases often excludes them from drug development programs. While individually rare, these diseases collectively affect more than 263 million individuals worldwide, and developing strategies to meet the needs of these patients is challenging (Vockley et al., Genet. Med. 25: 100022 (2023)).

[0008] Serum amyloid Al (SAAl) nucleic acid on human chromosome 11 encodes a member of the SSA1 family of apolipoproteins. The encoded preproprotein is proteolytically processed to generate the mature protein. This protein is a major acute phase protein that is highly expressed in response to inflammation and tissue injury. This protein also plays a role in high-density lipoprotein (HDL) metabolism and cholesterol homeostasis. High levels of this protein are associated with chronic inflammatory diseases including atherosclerosis, rheumatoid arthritis, Alzheimer's disease and Crohn's disease. This protein may also be a potential biomarker for certain PATENT 7328*158807PCT / NLM-1005-PC tumors. Alternate splicing results in multiple transcript variants that encode the same protein. A pseudogene of the SAA1 gene is found on chromosome 11.

[0009] Several variants of SAA1 nucleic acid have been identified (see, for example, World Wide Web Uniform Resource Locator (URL) ncbi.nlm.nih.gov / clinvar / ?term=SAAl[gene] and Sun et al., Gene 583(1): 48-57 (2016)). Ac.l01A> T pathogenic genetic variant in exon 3 of SAA1 nucleic acid, causing a p. Asp34Val modification in SAA1 protein, has been identified in only a few individuals worldwide. This particular pathogenic genetic variant has been associated with amyloid A amyloidosis.

[0010] SUMMARY

[0011] Provided in certain aspects are oligomeric compounds comprising a modified oligonucleotide that can reduce SAA1 expression. Also provided in certain aspects are compositions containing the oligomeric compounds. Provided also in certain aspects are methods, and uses of the oligomeric compounds, for reducing SAA1 expression, and in certain instances treating a SAA1 -associated medical condition. In certain aspects, provided are compositions, including products of manufacture and kits, and methods, for treating, preventing, reducing the symptoms or side effects of, or ameliorating a SAA1 -associated medical condition, such as amyloid A amyloidosis, for example. This Summary section is not limiting and certain aspects and embodiments of the technology are described further in the accompanying description, claim(s) and drawings.

[0012] The details of one or more exemplary embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. All publications, patents, patent applications cited herein are hereby expressly incorporated by reference in their entireties for all purposes.

[0013] DESCRIPTION OF DRAWINGS

[0014] The drawings illustrate certain embodiments of the technology and are not limiting. For clarity and ease of illustration, the drawings are not made to scale and, in some instances, various aspects can be shown exaggerated or enlarged to facilitate an understanding of particular embodiments. Like reference symbols in the various drawings indicate like elements. PATENT 7328*158807PCT / NLM-1005-PC FIG. 1 schematically illustrates a flow diagram showing an exemplary manufacturing process for a study drug, as described in Example 2.

[0015] FIG. 2 graphically illustrates patient plasma SAA1 levels, as described in Example 4.

[0016] FIG. 3 illustrates Table 8 showing clinical efficacy and safety laboratory values.

[0017] DETAILED DESCRIPTION

[0018] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0019] Definitions

[0020] Unless specific definitions are provided, the nomenclature used in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well-known and commonly used. Unless otherwise indicated, the following terms have the following meanings.

[0021] In alternative embodiments, “2’-deoxynucleoside” means a nucleoside including a 2’-H(H) deoxyribosyl sugar moiety. In certain embodiments, a 2’-dcoxynucl coside is a 2’-beta-D-deoxynucleoside and includes a 2’-beta-D-deoxyribosyl sugar moiety, which has the beta-D ribosyl configuration as found in naturally occurring deoxyribonucleic acids (DNA). In certain embodiments, a 2’-deoxynucleoside or a nucleoside including an unmodified 2’ -deoxyribosyl sugar moiety may include a modified nucleobase or may include an RNA nucleobase (uracil).

[0022] In alternative embodiments, “2’-M0E” means a 2’-OCH2CH2OCH3 group in place of the 2’ -OH group of a ribosyl sugar moiety. A “2’ -MOE sugar moiety” or a “2’-O-methoxyethyl sugar moiety” or “2’ -MOE ribosyl sugar moiety” means a sugar moiety with a 2’-OCH2CH2OCH3 group in place of the 2’ -OH group of a ribosyl sugar moiety. Unless otherwise indicated, a 2’ -MOE sugar moiety is in the beta-D configuration. “MOE” means O-methoxy ethyl. “2’-M0E nucleoside” or “2’-O(CH2)2OCH3 nucleoside” means a nucleoside including a 2’- MOE sugar moiety (or 2’-O(CH2)2OCH3 ribosyl sugar moiety).

[0023] In alternative embodiments, “2’-0Me” means a 2’-OCH3 group in place of the 2’ -OH group of a ribosyl sugar moiety. A“2’-O-methyl sugar moiety” means a sugar PATENT 7328*158807PCT / NLM-1005-PC moiety with a 2’-OCH3 group in place of the 2’ -OH group of a ribosyl sugar moiety. Unless otherwise indicated, a 2’-0Me has the beta-D ribosyl stereochemical configuration. “2’-0Me nucleoside” means a nucleoside including a 2’-0Me sugar moiety.

[0024] In alternative embodiments, “2’-F” means a 2’-fluoro group in place of the 2’-OH group of a ribosyl sugar moiety. A“2’-F sugar moiety” or “2’ -fluororibosyl sugar moiety” means a sugar moiety with a 2’-F group in place of the 2’ -OH group of a ribosyl sugar moiety. Unless otherwise indicated, a 2’-F has the beta-D ribosyl stereochemical configuration. “2’-F nucleoside” means a nucleoside including a 2’-F sugar moiety.

[0025] In alternative embodiments, “2 ’-substituted nucleoside” means a nucleoside including a 2 ’-substituted furanosyl sugar moiety. “2’ -substituted” in reference to a sugar moiety means a sugar moiety including at least one 2'- substituent group other than H or OH.

[0026] In alternative embodiments, “5-methylcytosine” means a cytosine modified with a methyl group attached to the 5 position. A 5-methylcytosine is a modified nucleobase.

[0027] In alternative embodiments, “abasic sugar moiety” means a sugar moiety of a nucleoside that is not attached to a nucleobase. Such abasic sugar moieties are sometimes referred to as “abasic nucleosides.”

[0028] In alternative embodiments, “administration” or “administering” means providing a pharmaceutical agent or composition to cells. “Administration” or “administering” can be providing a pharmaceutical agent or composition to cells of a subject, and can be providing a pharmaceutical agent or composition to a subject.

[0029] In alternative embodiments, “antisense activity” means any detectable and / or measurable change attributable to the hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity is a decrease in the amount or expression of a target nucleic acid or protein encoded by such target nucleic acid compared to target nucleic acid levels or target protein levels in the absence of the antisense compound.

[0030] In alternative embodiments, “antisense agent” means an antisense compound and optionally one or more additional features, such as a sense compound. PATENT 7328*158807PCT / NLM-1005-PC In alternative embodiments, “antisense compound” means an antisense oligonucleotide and optionally one or more additional features, such as a conjugate group.

[0031] In alternative embodiments, “sense compound” means a sense oligonucleotide and optionally one or more additional features, such as a conjugate group.

[0032] In alternative embodiments, “antisense oligonucleotide” means an oligonucleotide, including the oligonucleotide portion of an antisense compound, that is capable of hybridizing to a target nucleic acid and is capable of at least one antisense activity. Antisense oligonucleotides include but are not limited to antisense RNase H oligonucleotides and antisense RNAi oligonucleotides.

[0033] In alternative embodiments, “sense oligonucleotide” means an oligonucleotide, including the oligonucleotide portion of a sense compound, that is capable of hybridizing to an antisense oligonucleotide. Sense oligonucleotides include, but are not limited to, sense RNAi oligonucleotides.

[0034] In alternative embodiments, “bicyclic nucleoside” or “BNA” means a nucleoside including a bicyclic sugar moiety.

[0035] In alternative embodiments, “bicyclic sugar” or “bicyclic sugar moiety” means a modified sugar moiety including two rings, where the second ring is formed via a bridge connecting two of the atoms in the first ring thereby forming a bicyclic structure. In certain embodiments, the first ring of the bicyclic sugar moiety is a furanosyl sugar moiety. In certain embodiments, the furanosyl sugar moiety is a ribosyl sugar moiety. In certain embodiments, the bicyclic sugar moiety does not include a furanosyl sugar moiety.

[0036] In alternative embodiments, “blunt” or “blunt ended” in reference to an oligomeric duplex formed by two oligonucleotides means that there are no terminal unpaired nucleotides (i.e., no overhanging nucleotides). One or both ends of a doublestranded RNAi agent can be blunt.

[0037] In alternative embodiments, “cell-targeting moiety” means a conjugate group or portion of a conjugate group that is capable of binding to a particular cell type or particular cell types.

[0038] In alternative embodiments, “cerebrospinal fluid” or “CSF” means the fluid filling the space around the brain and spinal cord. “Artificial cerebrospinal fluid” or “aCSF” means a prepared or manufactured fluid that has certain properties (for PATENT 7328*158807PCT / NLM-1005-PC example, osmolarity, pH, and / or electrolytes) similar to cerebrospinal fluid and is biocompatible with CSF.

[0039] In alternative embodiments, “chirally enriched” in reference to a population means a plurality of molecules of identical molecular formula, where the number or percentage of molecules within the population that contain a particular stereochemical configuration at a particular chiral center is greater than the number or percentage of molecules expected to contain the same particular stereochemical configuration at the same particular chiral center within the population if the particular chiral center were stereo-random. Chirally enriched populations of molecules having multiple chiral centers within each molecule may contain one or more stereo-random chiral centers. In certain embodiments, the molecules are modified oligonucleotides. In certain embodiments, the molecules are oligomeric compounds including modified oligonucleotides. In certain embodiments, the chiral center is at the phosphorous atom of a phosphorothioate internucleoside linkage. In certain embodiments, the chiral center is at the phosphorous atom of a mesyl phosphoramidate internucleoside linkage.

[0040] In alternative embodiments, “cleavable moiety” means a bond or group of atoms that is cleaved under physiological conditions, for example, inside a cell, an animal, or a human.

[0041] In alternative embodiments, “complementary” in reference to an oligonucleotide means that at least 70% of the nucleobases of the oligonucleotide or one or more portions thereof and the nucleobases of another nucleic acid or one or more portions thereof are capable of hydrogen bonding with one another when the nucleobase sequence of the oligonucleotide and the other nucleic acid are aligned in opposing directions. “Complementary nucleobases” means nucleobases that are capable of forming hydrogen bonds with one another. Complementary nucleobase pairs include adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), 5 -methylcytosine (mC) and guanine (G). Certain modified nucleobases that pair with unmodified nucleobases or with other modified nucleobases are known. For example, inosine can pair with adenosine, cytosine, or uracil. Complementary oligonucleotides and / or nucleic acids need not have nucleobase complementarity at each nucleoside. Rather, some mismatches are tolerated. “Fully complementary” or “100% complementary ” in reference to an PATENT 7328*158807PCT / NLM-1005-PC oligonucleotide, or a portion thereof, means that the oligonucleotide, or portion thereof, is complementary to another oligonucleotide or nucleic acid at each nucleobase of the shorter of the two oligonucleotides, or at each nucleoside if the oligonucleotides are the same length. “Complementary region” in reference to a region of an oligonucleotide means that at least 70% of the nucleobases of that region and the nucleobases of another nucleic acid or one or more regions thereof are capable of hydrogen bonding with one another when the nucleobase sequence of the oligonucleotide and the other nucleic acid are aligned in opposing directions.

[0042] In alternative embodiments, “conjugate group” means a group of atoms that is directly attached to an oligonucleotide and confers at least one property to the resulting conjugated oligonucleotide. Conjugate groups include a conjugate moiety and a conjugate linker that attaches the conjugate moiety to the oligonucleotide.

[0043] “Conjugate linker” means a single bond or a group of atoms including at least one bond that connects a conjugate moiety to an oligonucleotide. “Conjugate moiety” means a group of atoms covalently bound to an oligonucleotide via a conjugate linker.

[0044] In alternative embodiments, "contiguous" in the context of an oligonucleotide refers to nucleosides, nucleobases, sugar moieties, or internucleoside linkages that are immediately adjacent to each other. For example, “contiguous nucleobases” means nucleobases that are immediately adjacent to each other in a sequence and “contiguous nucleosides” means nucleosides that are immediately adjacent to each other in a sequence.

[0045] In alternative embodiments, “constrained ethyl” or “cEf ’ or “cEt sugar moiety” means a beta-D ribosyl bicyclic sugar moiety where the second ring of the bicyclic sugar is formed via a bridge connecting the 4’-carbon and the 2’-carbon of the beta-D ribosyl sugar moiety, where the bridge has the formula 4'-CH(CH3)-O-2', and where the methyl group of the bridge is in the S configuration. The term “cEt nucleoside” means a nucleoside including a cEt sugar moiety.

[0046] In alternative embodiments, “deoxy region” means a region of 5-12 contiguous nucleosides, where at least 70% of the nucleosides are 2’-beta-D-deoxynucleosides. In certain embodiments, each nucleoside is selected from a 2’-beta-D-deoxynucleoside, a bicyclic nucleoside, and a 2 ’-substituted nucleoside. In certain embodiments, a deoxy region supports RNase H activity. In certain embodiments, a deoxy region is the gap or internal region of a gapmer. PATENT 7328*158807PCT / NLM-1005-PC In alternative embodiments, “diluent” means an ingredient in a composition that lacks pharmacological activity but is pharmaceutically necessary or desirable. For example, a diluent in an injected composition can be a liquid, for example, aCSF, phosphate buffered saline (PBS), or saline solution.

[0047] In alternative embodiments, “double-stranded” in reference to a region or an oligonucleotide means a duplex formed by complementary strands of nucleic acids (including, but not limited to, oligonucleotides) hybridized to one another. In certain embodiments, the two strands of a double-stranded region are separate molecules. In certain embodiments, the two strands are regions of the same molecule that has folded onto itself (for example, a hairpin structure).

[0048] In alternative embodiments, “duplex” or “duplex region” means the structure formed by two oligonucleotides or portions thereof that are hybridized to one another.

[0049] In alternative embodiments, “gapmer” means a modified oligonucleotide including an internal region having a plurality of nucleosides that support RNase H cleavage positioned between external regions having one or more nucleosides, where the nucleosides in the internal region are chemically distinct from the nucleoside or nucleosides in the external regions. The internal region can be referred to as the “gap” and the external regions can be referred to as the “wings” or “wing segments.” In certain embodiments, the internal region is a deoxy region. The positions of the internal region or gap refer to the order of the nucleosides of the internal region and are counted starting from the 5 ’-end of the internal region. Unless otherwise indicated, “gapmer’ refers to a sugar motif. In certain embodiments, each nucleoside of the gap is a 2’-beta-D-deoxynucleoside. In certain embodiments, the gap includes one 2 ’-substituted nucleoside at position 1, 2, 3, 4, or 5 of the gap, and the remainder of the nucleosides of the gap are 2’-beta-D-deoxynucleosides. the term “MOE gapmer” indicates a gapmer having a gap including 2’-beta-D-deoxynucleosides and wings including 2’-M0E nucleosides, the term “mixed wing gapmer” indicates a gapmer having wings including modified nucleosides including at least two different sugar modifications. Unless otherwise indicated, a gapmer may include one or more modified internucleoside linkages and / or modified nucleobases and such modifications do not necessarily follow the gapmer pattern of the sugar modifications.

[0050] In alternative embodiments, “hybridization” means the annealing of oligonucleotides and / or nucleic acids. While not limited to a particular mechanism, PATENT 7328*158807PCT / NLM-1005-PC the most common mechanism of hybridization involves hydrogen bonding, which can be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense compound and a nucleic acid target. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an oligonucleotide and a nucleic acid target.

[0051] In alternative embodiments, “intemucleoside linkage” means the covalent linkage between contiguous nucleosides in an oligonucleotide. “Modified internucleoside linkage” means any internucleoside linkage other than a phosphodiester internucleoside linkage. “Phosphorothioate internucleoside linkage” or “PS intemucleoside linkage” is a modified internucleoside linkage in which one of the non-bridging oxygen atoms of a phosphodiester intemucleoside linkage is replaced with a sulfur atom. “Inverted nucleoside” means a nucleotide having a 3’ to 3’ and / or 5’ to 5’ intemucleoside linkage, as shown herein.

[0052] In alternative embodiments, “inverted sugar moiety” means the sugar moiety of an inverted nucleoside or an abasic sugar moiety having a 3’ to 3’ and / or 5’ to 5’ intemucleoside linkage.

[0053] In alternative embodiments, “linked nucleosides” are nucleosides that are connected in a contiguous sequence, where no additional nucleosides are presented between those that are linked.

[0054] In alternative embodiments, “linker-nucleoside” means a nucleoside that links, either directly or indirectly, an oligonucleotide to a conjugate moiety. Linker-nucleosides are located within the conjugate linker of an oligomeric compound.

[0055] Linker-nucleosides are not considered part of the oligonucleotide portion of an oligomeric compound even if they are contiguous with the oligonucleotide.

[0056] In alternative embodiments, “mismatch” or “non-complementary” means a nucleobase of a first nucleobase sequence that is not complementary with the corresponding nucleobase of a second nucleobase acid sequence or target nucleic acid when the first and second nucleobase sequences are aligned in opposing directions.

[0057] In alternative embodiments, “motif’ means a pattern of unmodified and / or modified sugar moieties, nucleobases, and / or internucleoside linkages, in an oligonucleotide. PATENT 7328*158807PCT / NLM-1005-PC In alternative embodiments, “non-bicyclic modified sugar moiety” means a modified sugar moiety that includes a modification, such as a substituent, that does not form a bridge between two atoms of the sugar to form a second ring.

[0058] In alternative embodiments, "nucleobase" means an unmodified nucleobase or a modified nucleobase. An “unmodified nucleobase” is adenine (A), thymine (T), cytosine (C), uracil (U), or guanine (G). A “modified nucleobase” is a group of atoms other than unmodified A, T, C, U, or G capable of pairing with at least one unmodified nucleobase. A“5-methylcytosine” is a modified nucleobase. A universal base is a modified nucleobase that can pair with any one of the five unmodified nucleobases. “Nucleobase sequence” means the order of contiguous nucleobases in a nucleic acid or oligonucleotide, including such nucleobases that are each optionally independently modified or unmodified, and independent of any sugar or internucleoside linkage modification. “Nucleobase sequence of’ a reference SEQ ID NO, refers only to the nucleobase sequence provided in such SEQ ID NO and therefore, does not limit sugar or intemucleoside linkage modifications. Unless otherwise specified, each nucleobase can be an unmodified nucleobase, or a modified nucleobase as defined herein. For example, “A” represents unmodified or modified adenine; “C” represents unmodified or modified cytosine, “T” represents unmodified or modified thymidine, “U” represents unmodified or modified uracil, and “G” represents unmodified or modified guanine. Modified nucleobases that fall outside of these definitions are represented in the sequence by a different symbol, such as an “X”. “Nucleoside” means a compound, or fragment of a compound, including a nucleobase and a sugar moiety. The nucleobase and sugar moiety each independently are unmodified or modified.

[0059] In alternative embodiments, “modified nucleoside” means a nucleoside including a modified nucleobase and / or a modified sugar moiety. Modified nucleosides include abasic nucleosides, which lack a nucleobase. “Linked nucleosides” are nucleosides that are connected in a contiguous sequence, where no additional nucleosides are presented between those that are linked. “Oligomeric agent” means an oligomeric compound and optionally one or more additional features, such as a second oligomeric compound. An oligomeric agent can be a singlestranded oligomeric compound or can be an oligomeric duplex formed by two complementary oligomeric compounds. PATENT 7328*158807PCT / NLM-1005-PC In alternative embodiments, “oligomeric compound” means an oligonucleotide and optionally one or more additional features, such as a conjugate group or terminal group. An oligomeric compound can be paired with a second oligomeric compound that is complementary to the first oligomeric compound or can be unpaired. A “singled-stranded oligomeric compound” is an unpaired oligomeric compound.

[0060] In alternative embodiments, “oligomeric duplex” means a duplex formed by two oligomeric compounds having complementary nucleobase sequences. Each oligomeric compound of an oligomeric duplex can be referred to as a “duplexed oligomeric compound.”

[0061] In alternative embodiments, “oligonucleotide” means a strand of linked nucleosides connected via internucleoside linkages, where each nucleoside and internucleoside linkage can be modified or unmodified. Unless otherwise indicated, oligonucleotides consist of 8-50 linked nucleosides. “Modified oligonucleotide” means an oligonucleotide, where at least one nucleoside or internucleoside linkage is modified. “Unmodified oligonucleotide” means an oligonucleotide that does not include any nucleoside modifications or internucleoside modifications. An oligonucleotide can be paired with a second oligonucleotide that is complementary to the oligonucleotide or it can be unpaired. A “single-stranded oligonucleotide” is an unpaired oligonucleotide. A “double-stranded oligonucleotide” is an oligonucleotide that is paired with a second oligonucleotide.

[0062] In alternative embodiments, “pharmaceutically acceptable carrier” and “pharmaceutically acceptable diluent” means any substance suitable for use in administering to a subject. Certain carriers and diluents enable pharmaceutical compositions to be formulated as, for example, tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspension and lozenges for the oral ingestion by a subject. In certain embodiments, a pharmaceutically acceptable carrier or diluent is sterile water, sterile saline, sterile buffer solution or sterile artificial cerebrospinal fluid.

[0063] In alternative embodiments, “pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of compounds.

[0064] Pharmaceutically acceptable salts typically retain a desired biological activity of the parent compound and do not impart undesired toxicological effects thereto. PATENT 7328*158807PCT / NLM-1005-PC In alternative embodiments, “pharmaceutical composition” means a mixture of substances suitable for administration to a subject. For example, a pharmaceutical composition may include an oligomeric compound and a sterile aqueous solution. In certain embodiments, a pharmaceutical composition shows activity in free uptake assay in certain cell lines.

[0065] In alternative embodiments, “population” with respect to molecules means a plurality of molecules of identical molecular formula. “Population” with respect to genetic variations refers to a plurality of individuals of the same species (humans for example).

[0066] In alternative embodiments, “prodrug” means a therapeutic agent in a first form outside the body that is converted to a second form within a subject or cells thereof. Typically, conversion of a prodrug within the subject is facilitated by the action of an enzyme (for example, endogenous or viral enzyme) or chemicals present in cells or tissues and / or by physiologic conditions. In certain embodiments, the first form of the prodrug is less active than the second form.

[0067] In alternative embodiments, “reducing” or “inhibiting” an amount or activity refers to a reduction or blockade of the transcriptional expression or activity relative to the transcriptional expression or activity in an untreated or control sample.

[0068] “Reducing” and “inhibiting” do not necessarily indicate a total elimination of transcriptional expression or activity.

[0069] In alternative embodiments, “selectively reducing” or “selectively inhibiting” an amount or activity refers to relative reduction or blockade of gene expression or activity of one species relative to another species. The different species can be different alleles or allelic variants, and can be a mutant allele relative to a wild-type allele in certain embodiments. A compound that selectively reduces expression of a first allele relative to a second allele reduces expression of the first allele 2-fold or more relative to expression of the second allele. Stated another way, an expression level of a first allele is at least 2-fold lower than the expression level of a second allele in the presence of an allele-selective compound. A compound that does not selectively reduce expression of a first allele relative to a second allele reduces expression of the first allele less than 2-fold relative to expression of the second allele. Stated another way, an expression level of a first allele is less than 2-fold lower than the expression level of a second allele in the presence of a non-allele-selective compound. In certain PATENT 7328*158807PCT / NLM-1005-PC embodiments, selective inhibition by an oligomeric compound is assessed by a selectivity ratio as described herein.

[0070] In alternative embodiments, “RNA” means an RNA transcript and includes pre-mRNA and mature mRNA unless otherwise specified. “RNAi agent” means an antisense agent that acts, at least in part, through RISC or Ago2 to modulate a target nucleic acid and / or protein encoded by a target nucleic acid. RNAi agents include, but are not limited to double-stranded siRNA, single-stranded RNA (ssRNAi), and microRNA, including microRNA mimics. RNAi agents may include conjugate groups and / or terminal groups. In certain embodiments, an RNAi agent modulates the amount, activity, and / or splicing of a target nucleic acid. The term RNAi agent excludes antisense agents that act through RNase H.

[0071] In alternative embodiments, “RNase H agent” means an antisense agent that acts through RNase H to modulate a target nucleic acid and / or protein encoded by a target nucleic acid. In certain embodiments, RNase H agents are single-stranded. In certain embodiments, RNase H agents are double-stranded. RNase H agents may include conjugate groups and / or terminal groups. In certain embodiments, an RNase H agent modulates the amount and / or activity of a target nucleic acid. The term “RNase H agent” excludes antisense agents that act principally through RISC / Ago2.

[0072] In alternative embodiments, “antisense RNase H oligonucleotide” means an oligonucleotide including a region that is complementary to a target sequence, and which includes at least one chemical modification suitable for RNase H-mediated nucleic acid reduction.

[0073] In alternative embodiments, “antisense RNAi oligonucleotide” means an oligonucleotide including a region that is complementary to a target sequence, and which includes at least one chemical modification suitable for RNAi-mediated nucleic acid reduction.

[0074] In alternative embodiments, “self-complementary” in reference to an oligonucleotide means an oligonucleotide that at least partially hybridizes to itself.

[0075] In alternative embodiments, “single-stranded” means a nucleic acid, including but not limited to an oligonucleotide, that is unpaired and is not part of a duplex. Single-stranded compounds are capable of hybridizing with complementary nucleic acids to form duplexes, at which point they are no longer single-stranded. PATENT 7328*158807PCT / NLM-1005-PC In alternative embodiments, “stabilized phosphate group” means a 5’-phosphate analog that is metabolically more stable than a 5 ’-phosphate as naturally occurs on DNA or RNA.

[0076] In alternative embodiments, “stereo-random” and “stereo-random chiral center” in the context of a population of molecules of identical molecular formula refer to a chiral center that is not controlled during synthesis, or enriched following synthesis, for a particular absolute stereochemical configuration. The stereochemical configuration of a chiral center is considered random when it is the result of a synthetic method that is not designed to control the stereochemical configuration. For example, in a population of molecules including a stereo-random chiral center, the number of molecules having the (S) configuration of the stereo-random chiral center can be but is not necessarily the same as the number of molecules having (R) configuration of the stereo-random chiral center (“racemic”). In certain embodiments, the stereo-random chiral center is not racemic because one absolute configuration predominates following synthesis, for example, due to the action of non-chiral reagents near the enriched stereochemistry of an adjacent sugar moiety. In certain embodiments, a stereo-random chiral center is a stereo-random phosphorothioate internucleoside linkage or a mesyl phosphoramidate internucleoside linkage.

[0077] In alternative embodiments, “subject” means a human or non-human animal. In certain embodiments, the subject is a human. “Animal” means a human or non-human animal.

[0078] In alternative embodiments, “sugar moiety” means an unmodified sugar moiety or a modified sugar moiety. “Unmodified sugar moiety” means a 2’-0H(H) beta-D-ribosyl sugar moiety, as found in RNA (an “unmodified RNA sugar moiety”), or a 2’-H(H) beta-D-deoxyribosyl sugar moiety, as found in DNA (an “unmodified DNA sugar moiety”). Unmodified sugar moi eties have one hydrogen at each of the 1’, 3’, and 4’ positions, an oxygen at the 3’ position, and two hydrogens at the 5’ position. “Modified sugar moiety” or “modified sugar” means a modified furanosyl sugar moiety or a sugar surrogate.

[0079] In alternative embodiments, "sugar surrogate" means a modified sugar moiety having other than a furanosyl moiety that can link a nucleobase to another group, such as an internucleoside linkage, conjugate group, or terminal group in an oligonucleotide. Modified nucleosides including sugar surrogates can be incorporated PATENT 7328*158807PCT / NLM-1005-PC into one or more positions within an oligonucleotide and such oligonucleotides are capable of hybridizing to complementary oligomeric compounds or target nucleic acids.

[0080] In alternative embodiments, “treating” means improving a subject’s medical condition by administering a pharmaceutical agent or composition, such as a pharmaceutical agent or composition comprising an oligomeric compound described herein for example. In certain embodiments, treating a subject improves a symptom or hallmark relative to the same symptom or hallmark in the absence of the treatment. In certain embodiments, treatment reduces in the severity or frequency of a symptom or hallmark, or delays the onset of a symptom or hallmark, slows the progression of a symptom or hallmark, or slows the severity or frequency of a symptom or hallmark.

[0081] In alternative embodiments, “therapeutically effective amount” means an amount of a pharmaceutical agent or composition that provides a therapeutic benefit to a subject. For example, a therapeutically effective amount improves a symptom of a disease.

[0082] In alternative embodiments, “medical condition” means a disease, disorder or other condition for which a subject receives medical attention, medical service, treatment, diagnosis, consultation or a drug prescription. A medical condition can be a SAA1 -associated medical condition, and in certain embodiments, is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia.

[0083] In alternative embodiments, “symptom or hallmark” means a physical feature or test result that indicates the existence or extent of a medical condition. In certain embodiments, a symptom is apparent to a subject or to a medical professional examining or testing the subject. In certain embodiments, a hallmark is a particular indicator, such as a molecular indicator for example. In certain embodiments, a hallmark is an indicator that can be identified by testing. In certain embodiments, a hallmark is identified by diagnostic testing. In certain embodiments, a hallmark is identified by invasive testing, such as a post-mortem test for example. In certain embodiments, a symptom or hallmark is one or more of proteinuria, nephrotic syndrome, low blood protein level, low blood albumin level, edema, high cholesterol, enlarged spleen, enlarged liver, enlarged thyroid, orthostatic hypotension, gastrointestinal atony, diarrhea and constipation. PATENT 7328*158807PCT / NLM-1005-PC In alternative embodiments, “ameliorate” with reference to a treatment means improvement in at least one symptom or hallmark relative to the same symptom or hallmark in the absence of the treatment. In certain embodiments, amelioration is reduction in severity or frequency of a symptom or hallmark or delayed onset or slowing of progression in severity or frequency of a symptom or hallmark.

[0084] Progression or severity of a symptom or hallmark can be determined by known subjective or objective measures.

[0085] In alternative embodiments, “terminal group” means a chemical group or group of atoms that is covalently linked to a terminus of an oligonucleotide.

[0086] In alternative embodiments, “Target nucleic acid,” which includes “target RNA,” means a nucleic acid that an antisense compound is designed to affect. Target RNA means an RNA transcript and includes pre-mRNA and mature mRNA unless otherwise specified. “Target region” means a portion of a target nucleic acid to which an oligomeric compound is designed to hybridize.

[0087] In alternative embodiments, “targeted to” means an oligomeric compound designed to hybridize to a target region of a SAA1 nucleic acid. A modified oligonucleotide of a oligomeric compound targeted to a SAA1 nucleic acid can consist of a nucleobase sequence that is at least 80% complementary to, or at least 85% complementary to, or at least 90% complementary to, or at least 95% complementary to, or 100% complementary to, an equal length target region portion of a SAA1 nucleic acid. An “equal length portion” and “equal length target region portion” means a portion of contiguous nucleobases within a SAA1 nucleic acid nucleobase sequence, for example within SEQ ID NO:1 or SEQ ID NO:2. In certain embodiments, an oligomeric compound oligonucleotide is 100% complementary to an equal length target region portion of a SAA1 nucleic acid except for a one-nucleobase mismatch.

[0088] In alternative embodiments, “allele” refers to one member of a pair of genes or one member of a series of different forms of a DNA sequence that can exist at a single locus or marker on a specific chromosome. For a diploid organism or cell or for autosomal chromosomes, each allelic pair normally occupies corresponding positions (loci) on a pair of homologous chromosomes, one inherited from the mother and one inherited from the father. If these alleles are identical, the organism or cell is “homozygous” for that allele. If the alleles differ, the organism or cell is PATENT 7328*158807PCT / NLM-1005-PC “heterozygous” for that allele. A“SAA1 allele” refers to one member of a pair of SAA1 genes on a specific chromosome. “Mutant allele” refers to an allele typically containing a pathogenic genetic variant associated with the medical condition. For example, a SAA1 allele containing a pathogenic genetic variant associated with a medical condition is referred to as a “mutant SAA1 allele” or a “mutant SAA1 nucleic acid.” “Wild type allele” refers to an allele not containing a pathogenic genetic variant present in a mutant allele. For example, a SAA1 allele not containing a pathogenic genetic variant associated with a medical condition is referred to as a “wild-type SAA1 allele” or “wild-type SAA1 nucleic acid.” Each of two SAA1 alleles that differ from one another can be referred to as a “SAA1 allelic variant.” “Allelic variant nucleic acid” refers to a chromosome nucleic acid or transcription product nucleic acid such as a pre-mRNA or mRNA corresponding to a particular allele. For example, “mutant SAA1 allele nucleic acid” and “wild-type SAA1 allele nucleic acid” each independently refers to a chromosome nucleic acid or transcription product nucleic acid such as pre-mRNA or mRNA corresponding to the particular allele, for example. A “mutant SAA1 nucleic acid” and a “wild-type SAA1 nucleic acid” refers to a chromosome nucleic acid or transcription product nucleic acid such as a pre-mRNA or mRNA.

[0089] In alternative embodiments, “genetic variation” means a variation of one or more nucleobases between allelic variants of an individual and / or between genomes of individuals of the same species at a particular polymorphic position. A genetic variation can include two or more genetic variants at the polymorphic position. A genetic variation generally occurs at a “genetic variation position” within a “genetic variation site.” “Genetic variation position” refers to the nucleobase position(s) of the polymorphic genetic variation on a reference nucleobase sequence within a genetic variation site. “Genetic variation site” refers to the genetic variation position and linked nucleobases on one or both sides (5’ side and / or 3’ side) of the genetic variation position, which can be contained in a target region to which an oligomeric compound is targeted. A genetic variant associated with a medical condition is referred to as a “pathogenic genetic variant” at a pathogenic genetic variation position and / or site.

[0090] In alternative embodiments, “pathogenic genetic variation site” means a genetic variation site comprising a pathogenic genetic variant present on a mutant allele nucleic acid. A pathogenic genetic variant typically is associated with a medical PATENT 7328*158807PCT / NLM-1005-PC condition and can cause the medical condition. A pathogenic variation site can be a differentiating variation site targeted by an oligomeric compound. A pathogenic genetic variation site can include a polymorphic single nucleotide variation position, for which a pathogenic genetic variant includes a particular nucleobase at the pathogenic genetic variation position. A pathogenic genetic variant can be a gain-of-function (GOF) genetic variant, which typically results in a mutant protein activity not observed for wild-type protein. A GOF genetic variant can be a toxic GOF (TGOF) genetic variant in certain instances. A pathogenic genetic variant of a pathogenic genetic variation position typically is not depicted in a reference sequence (for example, a reference genomic sequence of SEQ ID NO: 1 or a reference cDNA sequence of SEQ ID NO:2). A pathogenic allelic variant can be heterozygous, present on a mutant allele and not on a wild-type allele, in certain embodiments.

[0091] In alternative embodiments, “genotype” means an identification of one or more nucleobases in genomic nucleic acid at a particular genetic location. The particular location or site can include or be a genetic variation position or genetic variation site. A genotype can be identified in nucleic acid from an individual or group of individuals. A genotype can be a nucleobase, nucleobase sequence or symbol representative of the nucleobase or nucleobases. A genotype can be determined in situ or for sample nucleic acid.

[0092] In alternative embodiments, “sample nucleic acid” means nucleic acid from a cell, tissue or subject, and can be nucleic acid isolated from cell, tissue or subject. A sample from a subject can be a fluid or solid sample in certain instances.

[0093] Exemplary Embodiments

[0094] Non-limiting embodiments of the technology are described hereafter in a numbered format starting with embodiment Al (for example, “Al” is “embodiment Al”).

[0095] Al. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to an equal length portion of a SAA1 nucleic acid, and wherein the modified oligonucleotide comprises at least one modification selected from a modified sugar moiety and a modified internucleoside linkage. PATENT 7328*158807PCT / NLM-1005-PC A2. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and comprising or consisting of a nucleobase sequence containing at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of any of (i) SEQ ID NO:4 to SEQ ID NO: 12, (ii) SEQ ID NO:4 to SEQ ID NO:7 and SEQ ID NO:9 to SEQ ID NO: 11, (iii) SEQ ID NO:8, (iv) SEQ ID NO: 12 or (v) SEQ ID NO:4, wherein the modified oligonucleotide comprises at least one modification selected from a modified sugar moiety and a modified internucleoside linkage.

[0096] A3. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and comprising or consisting of a nucleobase sequence containing at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases complementary to:

[0097] an equal length portion of nucleobases 3458-3708, 3624-3727 or 3644-3707 of SEQ ID NO: 1; or

[0098] an equal length portion of nucleobases 3644-3663, 3647-3666, 3652-3671, 3663-3682, 3666-3685, 3677-3696 or 3688-3707 of SEQ ID NO: 1; or an equal length portion of nucleobases 3643-3702, 3653-3692 or 3663-3682 of SEQ ID NO: 1; or

[0099] an equal length portion of nucleobases 720-3074, 2908-2967 or 2928-2947 of SEQ ID NO: 1; or

[0100] an equal length portion of nucleobases 720-2935, 2200-2259 or 2220-2239 of SEQ ID NO: 1;

[0101] wherein the modified oligonucleotide comprises at least one modification selected from a modified sugar moiety and a modified internucleoside linkage. A4. The oligomeric compound of any of embodiments Al -A3, wherein the modified oligonucleotide comprises or consists of a nucleobase sequence that is at least 85%, at least 90%, at least 95%, or 100% complementary to an equal length portion of a SAA1 nucleic acid, wherein the SAA1 nucleic acid has the nucleobase sequence of SEQ ID NO: 1 or SEQ ID NO:2.

[0102] A5. The oligomeric compound of any of embodiments A1-A4, wherein the modified oligonucleotide consists of 12 to 20, 12 to 25, 12 to 30, 12 to 50, 13 to 20, 13 to 25, 13 to 30, 13 to 50, 14 to 20, 14 to 25, 14 to 30, 14 to 50, 15 to 20, 15 to 25, 15 to 30, PATENT 7328*158807PCT / NLM-1005-PC 15 to 50, 16 to 18, 16 to 20, 16 to 25, 16 to 30, 16 to 50, 17 to 20, 17 to 25, 17 to 30, 17 to 50, 18 to 20, 18 to 25, 18 to 30, 18 to 50, 19 to 20, 19 to 25, 19 to 30, 19 to 50, 20 to 25, 20 to 30, or 20 to 50 linked nucleosides.

[0103] A6. The oligomeric compound of any one of embodiments A1-A5, which is not SAA1 allele selective.

[0104] Bl. The oligomeric compound of any one of embodiments A1-A6, wherein the modified oligonucleotide comprises at least one modified nucleoside.

[0105] B2. The oligomeric compound of embodiment Bl, wherein the at least one modified nucleoside comprises a modified sugar.

[0106] B3. The oligomeric compound of embodiment B2, wherein the modified sugar comprises a bicyclic sugar.

[0107] B4. The oligomeric compound of embodiment B3, wherein the bicyclic sugar comprises a 2’ -4’ bridge selected from -O-CH2- and -O-CH(CH3)-.

[0108] B5. The oligomeric compound of embodiment Bl, wherein the at least one modified nucleoside comprises a non-bicyclic modified sugar.

[0109] B6. The oligomeric compound of embodiment B5, wherein the non-bicyclic modified sugar moiety is a 2 ’-substituted sugar moiety or a 5 ’-substituted sugar moiety.

[0110] B7. The oligomeric compound of embodiment B6, wherein the 2’-substitued sugar moiety is a 2'-MOE sugar moiety, a 2’-OMe sugar moiety or a 2’-F sugar moiety; or the 5 ’-substituted sugar moiety is a 5 ’-methyl modified sugar moiety or 5 ’-ethyl modified sugar moiety.

[0111] B8. The oligomeric compound of embodiment Bl, wherein the at least one modified nucleoside comprises a sugar surrogate.

[0112] B9. The oligomeric compound of embodiment B8, wherein the sugar surrogate is selected from morpholino, modified morpholino, glycol nucleic acid (GNA), sixmembered tetrahydropyran (THP), and F-hexitol nucleic acid (F-HNA).

[0113] B10. The oligomeric compound of any one of embodiments A1-B9, wherein the modified oligonucleotide is a gapmer.

[0114] Bll. The oligomeric compound of any one of embodiments Al -B10, wherein the modified oligonucleotide comprises at least one modified internucleoside linkage. B12. The oligomeric compound of embodiment Bll, wherein the at least one modified internucleoside linkage is a phosphorothioate intemucleoside linkage. PATENT 7328*158807PCT / NLM-1005-PC B13. The oligomeric compound of any one of embodiments A1-B12, wherein the modified oligonucleotide comprises at least one phosphodiester internucleoside linkage.

[0115] B14. The oligomeric compound of any one of embodiments B13, wherein the modified oligonucleotide comprises phosphodiester internucleoside linkages and a phosphorothioate internucleoside linkages.

[0116] Bl 5. The oligomeric compound of any one of embodiments B11-B14, wherein the modified oligonucleotide comprises at least one mesyl phosphoramidate linkage. Bl 6. The oligomeric compound of any one of embodiments B11-B15, wherein the modified oligonucleotide comprises at least one busyl phosphoramidate linkage. Bl 7. The oligomeric compound of any one of embodiments B11-B16, wherein at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, or 19 internucleoside linkages of the modified oligonucleotide are phosphorothioate internucleoside linkages.

[0117] Bl 8. The oligomeric compound of any one of embodiments B11-B17, wherein the modified oligonucleotide comprises an intemucleoside linkage motif (from 5' to 3’) selected from sososssssssssssooss, sooosssssssssssooss, soooossssssssssooss or sssssssssssssssssss, wherein each “s” represents a phosphorothioate intemucleoside linkage and each “o” represents a phosphodiester intemucleoside linkage, and optionally each “s” represents a (Sp)-phosphorothioate intemucleoside linkage.

[0118] Bl 9. The oligomeric compound of any one of embodiments Al -Bl 8, wherein the modified oligonucleotide comprises at least one modified nucleobase.

[0119] B20. The oligomeric compound of embodiment Bl 9, wherein the modified nucleobase is a 5-methylcytosine.

[0120] B21. The oligomeric compound of embodiment B20, wherein each cytosine is a 5-methylcytosine.

[0121] B22. The oligomeric compound of any one of embodiments Al -B21, comprising a deoxy region consisting of two or more contiguous nucleosides each comprising a 2'-beta-deoxyribosyl sugar moiety.

[0122] B23. The oligomeric compound of embodiment B22, wherein the deoxy region consists of 3, 4, 5, 6, 7, 8, 9, 10, or 3-10, contiguous nucleosides, each comprising a 2'-beta-deoxyribosyl sugar moiety. PATENT 7328*158807PCT / NLM-1005-PC B24. The oligomeric compound of embodiment B22 or B23, wherein each nucleoside immediately adjacent to the deoxy region comprises a modified sugar.

[0123] B25. The oligomeric compound of any one of embodiments A1-B25, wherein the modified oligonucleotide comprises:

[0124] a 5’-region consisting of 1-6 linked 5’-region nucleosides;

[0125] a central region consisting of 6-10 linked central region nucleosides; and a 3 ’-region consisting of 1-6 linked 3 ’-region nucleosides; wherein:

[0126] each of the 5 ’-region nucleosides and each of the 3 ’-region nucleosides comprises a modified sugar moiety, and three or more of the central region nucleosides comprise a 2’-beta-D-deoxyribosyl sugar moiety.

[0127] B26. The oligomeric compound of any one of embodiments A1-B25, wherein the modified oligonucleotide comprises:

[0128] a 5 ’-region consisting of 5 linked 5 ’-region nucleosides;

[0129] a central region consisting of 10 linked central region nucleosides; and a 3 ’-region consisting of 5 linked 3 ’-region nucleosides; wherein:

[0130] each of the 5 ’-region nucleosides and each of the 3 ’-region nucleosides comprises a modified sugar moiety, and three or more of the central region nucleosides comprise a 2’-beta-deoxyribosyl sugar moiety.

[0131] B27. The oligomeric compound of embodiment B25 or B26, wherein each of the 5’-region nucleosides and each of the 3’-region nucleosides comprises a 2 ’-substituted sugar moiety.

[0132] B28. The oligomeric compound of embodiment B27, wherein each of the 5 ’-region nucleosides and each of the 3 ’-region nucleosides comprises a 2’ -MOE sugar moiety. B29. The oligomeric compound of any one of embodiments B25-B28, wherein the central region comprises four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten central region nucleosides comprising a 2'-beta-deoxyribosyl sugar moiety.

[0133] B30. The oligomeric compound of any one of embodiments B25-B29, wherein the central region comprises 1, 2, 3, 4 or 5, or 1 to 5, modified sugar moieties.

[0134] B31. The oligomeric compound of embodiment B30, wherein one or more of the modified sugar moieties is a non-bicyclic modified sugar moiety.

[0135] B32. The oligomeric compound of embodiment B30 or B31, wherein one or more of the modified sugar moieties is a bicyclic modified sugar moiety. PATENT 7328*158807PCT / NLM-1005-PC B33. The oligomeric compound of any one of embodiments B30-B32, wherein one or more of the modified sugar moieties comprises a 2’ substituent.

[0136] B34. The oligomeric compound of embodiment B33, wherein one or more of the modified sugar moieties is a 2’-0Me modified sugar moiety.

[0137] B35. The oligomeric compound of any one of embodiments B30-B34, wherein one or more of the modified sugar moieties comprises a 5’ substituent.

[0138] B36. The oligomeric compound of embodiment B35, wherein one or more of the modified sugar moieties is a 5 ’-methyl modified sugar moiety or 5 ’-ethyl modified sugar moiety.

[0139] B37. The oligomeric compound of any one of embodiments B25-B36, wherein:

[0140] the central region comprises a 5’ terminus; and

[0141] position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises a modified sugar moiety.

[0142] B38. The oligomeric compound of embodiment B37, wherein position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises a modified sugar moiety comprising a 2’ substituent.

[0143] B39. The oligomeric compound of embodiment B38, wherein the modified sugar moiety independently is a 2’-0Me modified sugar moiety.

[0144] B40. The oligomeric compound of any one of embodiments B25-B39, wherein:

[0145] the central region comprises a 5’ terminus; and

[0146] position 3, or position 4, or positions 3 and 4 of the central region, as counted from the 5' terminus of the central region, each independently comprises a modified sugar moiety.

[0147] B41. The oligomeric compound of embodiment B40, wherein position 3, or position 4, or positions 3 and 4 of the central region, as counted from the 5' terminus of the central region, each independently comprises modified sugar moiety comprising a 5’ substituent.

[0148] B42. The oligomeric compound of embodiment B41, wherein position 3, or position 4, or positions 3 and 4 of the central region, as counted from the 5' terminus of the central region, each independently comprises a 5 ’-methyl modified sugar moiety. PATENT 7328*158807PCT / NLM-1005-PC B43. The oligomeric compound of embodiment B42, wherein the 5 ’-methyl modified sugar moiety independently is a R-5’-methyl modified sugar moiety or a S-5’-methyl modified sugar moiety.

[0149] B44. The oligomeric compound of any one of embodiments B25-B43, wherein the central region comprises 1, 2, 3, 4 or 5, or 1 to 5, mesyl phosphoramidite and / or busyl phosphoramidite internucleoside linkages.

[0150] B45. The oligomeric compound of embodiment B44, wherein:

[0151] the central region comprises a 5’ terminus; and

[0152] position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises a mesyl phosphoramidite and / or a busyl phosphoramidite intemucleoside linkage.

[0153] B46. The oligomeric compound of any one of embodiments B25-B45, wherein the central region comprises 1, 2, 3, 4 or 5, 1 to 5, alkylphosphonate internucleoside linkages.

[0154] B47. The oligomeric compound of embodiment B46, wherein:

[0155] the central region comprises a 5’ terminus; and

[0156] position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises an alkylphosphonate intemucleoside linkage.

[0157] B48. The oligomeric compound of embodiment B46 or B47, wherein the alkylphosphonate intemucleoside linkage comprises methoxypropyl (MOP).

[0158] B49. The oligomeric compound of any one of embodiments B25-B48, wherein the central region comprises the deoxy region of any one of embodiments B22-B24. B50. The oligomeric compound of embodiment B49, wherein the modified oligonucleotide comprises a sugar motif of 5’-eeeeeddddddddddeeeee-3’, wherein each "d " represents a 2’-beta-D~deoxyribosyl sugar moiety, each “e” represents a 2’-MOE sugar moiety.

[0159] B51. The oligomeric compound of any one of embodiments A1-B50, wherein the modified oligonucleotide consists of 12-30, 12-22, 12-20, 14-18, 14-20, 15-17, 15-25, 16-20, 18-22, or 18-20 linked nucleosides, or a pharmaceutically acceptable salt thereof. PATENT 7328*158807PCT / NLM-1005-PC B52. The oligomeric compound of any one of embodiments A1-B51, wherein the modified oligonucleotide consists of 20 linked nucleosides, or a pharmaceutically acceptable salt thereof.

[0160] B53. The oligomeric compound of embodiment B51 or B52, which is a pharmaceutically acceptable salt comprising one or more cations selected from sodium, potassium, calcium and magnesium.

[0161] B54. The oligomeric compound of any one of embodiments A1-B53, wherein the modified oligonucleotide consists of 20 linked nucleosides.

[0162] B55. The oligomeric compound of any one of embodiments A1-B54, wherein the oligomeric compound consists of the modified oligonucleotide.

[0163] B56. The oligomeric compound of any one of embodiments A1-B54, wherein the oligomeric compound comprises a conjugate group.

[0164] B57. The oligomeric compound of embodiment B56, wherein the conjugate group comprises a conjugate moiety and a conjugate linker.

[0165] B58. The oligomeric compound of embodiment B57, wherein the conjugate linker is a phosphodiester linker.

[0166] B59. The oligomeric compound of embodiment B57, wherein the conjugate linker consists of a single bond.

[0167] B60. The oligomeric compound of any one of embodiments B57-B59, wherein the conjugate linker is cleavable.

[0168] B61. The oligomeric compound of any one of embodiments B57-B60, wherein the conjugate linker comprises 1-3 linker-nucleosides.

[0169] B62. The oligomeric compound of any one of embodiments B56-B61, wherein the conjugate group is attached to the modified oligonucleotide at the 5 ’-end of the modified oligonucleotide.

[0170] B63. The oligomeric compound of any one of embodiments B56-B62, wherein the conjugate group is attached to the modified oligonucleotide at the 3 ’-end of the modified oligonucleotide.

[0171] B64. The oligomeric compound of any one of embodiments B56-B63, wherein the conjugate group comprises a targeting moiety and a linker moiety.

[0172] B65. The oligomeric compound of embodiment B64, wherein the targeting moiety comprises a carbohydrate ligand. PATENT 7328*158807PCT / NLM-1005-PC B66. The oligomeric compound of embodiment B65, wherein the carbohydrate ligand has affinity for an asialoglycoprotein receptor (ASGPR).

[0173] B67. The oligomeric compound of embodiment B65 or B66, wherein the carbohydrate ligand can target the oligomeric compound to liver cells and / or hepatocytes.

[0174] B68. The oligomeric compound of any one of embodiments B65-B67, wherein the carbohydrate ligand comprises or consists of aN-acetylgalactosamine (GalNAc) moiety.

[0175] B69. The oligomeric compound of embodiment B68, wherein the targeting moiety comprises multiple GalNAc moieties, a branching group and a tether moiety linking the branching group to each of the GalNAc moieties.

[0176] B70. The oligomeric compound of embodiment B68 or B69, wherein the targeting moiety comprises two, three or four GalNAc moieties, and optionally three GalNAc moieties.

[0177] B71. The oligomeric compound of any one of embodiments B64-B70, wherein the targeting moiety is cleavable.

[0178] B72. The oligomeric compound of any one of embodiments B69-B71, wherein the branching group is cleavable.

[0179] B73. The oligomeric compound of any one of embodiments B64-B72, wherein: the targeting moiety comprises or consists of a structure according to the following general Formula A:

[0180]

[0181] Formula A PATENT 7328*158807PCT / NLM-1005-PC and each m and each n in Formula A independently is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

[0182] B74. The oligomeric compound of embodiment B73, wherein each m independently is an integer of 5, 6 or 7 and each n independently is an integer of 1, 2 or 3 in Formula A.

[0183] B75. The oligomeric compound of embodiment B73 or B74, wherein each m is the integer 6 and each n is the integer 2 in Formula A.

[0184] B76. The oligomeric compound of any one of embodiments B64-B75, wherein: the linker moiety comprises or consists of a structure according to one of the following general Formula B, Formula C or Formula D:

[0185]

[0186] and each n independently is zero or an integer of 1, 2, 3, 4, 5, 6 or 7 in Formula B or Formula C.

[0187] B77. The oligomeric compound of embodiment B76, wherein each n independently is an integer of 1, 2, 3, 4 or 5 in Formula B or Formula C.

[0188] B78. The oligomeric compound of embodiment B76 or B77, wherein each n independently is the integer 2 or 5 in Formula B or Formula C.

[0189] B79. The oligomeric compound of any one of embodiments B64-B78, wherein the targeting moiety and linker moiety together are according to the following general formula: PATENT 7328*158807PCT / NLM-1005-PC

[0190]

[0191] B80. The oligomeric compound of any one of embodiments B64-B79, wherein the linker moiety is joined to the oligonucleotide by a phosphodiester moiety.

[0192] B81. The oligomeric compound of embodiment B80, wherein the oligonucleotide compound comprises a 5’ terminal nucleoside and the phosphodiester moiety is joined to the 5’ end of the 5’ terminal nucleoside.

[0193] B82. The oligomeric compound of embodiment B81, wherein the 5' terminal nucleoside comprises a sugar moiety and a methylene group linked to the 5’ end of the sugar moiety, and the phosphodiester moiety is attached to the methylene group. B83. The oligomeric compound of any one of embodiments A1-B82, wherein the oligomeric compound comprises a terminal group.

[0194] B84. The oligomeric compound of any one of embodiments A1-B83, wherein the oligomeric compound does not comprise linker-nucleosides.

[0195] B85. An oligomeric compound comprising an oligonucleotide and conjugated group according to the following general formula:

[0196] 5'-GalNAc group-p-A-mC-mC-mC-A-T-T-G-T-G-T-A-mC-mC-mC-mU-mC-mU- mC-mC-3' (SEQ ID NO : 15)

[0197] wherein:

[0198] the oligonucleotide is A-mC-mC-mC-A-T-T-G-T-G-T-A-mC-mC-mC-mU- mC-mU-mC-mC (SEQ ID NO:16 containing designated nucleosides; each underlined nucleoside contains a ribose 2'-0 modified by a methoxy ethyl (MOE) group;

[0199] all internucleoside linkages are phosphorothioate linkages;

[0200] each uracil base and cytosine base is modified at the 5 position with a methyl group; PATENT 7328*158807PCT / NLM-1005-PC the conjugated group is the GalNAc group and the GalNAc group consists of a targeting moiety containing N-acetylgalactosamine moieties and a linker moiety joining the targeting moiety to the oligonucleotide;

[0201] p is a phosphodiester linkage between the GalNAc group and the oligonucleotide; and

[0202] the targeting moiety and the linker moiety optionally is according to any one of embodiments B65-B82.

[0203] B86. The oligomeric compound of embodiment B85, wherein the GalNAc group and a portion of the phosphodiester linkage comprise a structure according to the general formula:

[0204]

[0205] B87. An oligomeric compound of any one of embodiments B56-B86, comprising or consisting of a / / - -amZ>o-5'-O-{[(6-{5-[(tris{3-[6-(2-acetamido-2-deoxy-P-D-galactopyranosyloxy)hexylamino]-3-oxopropoxymethyl})methyl]amino-5-oxopentanamido}hexyl)]phospho}-2'-O-(2-methoxyethyl)-P-thioadenylyl-(3'-O— >5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3'-O—>5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3'-O— >5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3 '-O^ 5 '-O)-2'-O-(2-methoxyethyl)-P-thioadenylyl-(3 O—>5 '-O)-P-thiothymidylyl-(3 '-O—>5 '-O)-P-thiothymidylyl-(3 '-O—>5 '-O)-2'-deoxy- -thioguanylyl-(3'-O^5'-O)-P-thiothymidylyl-(3 '-( ^5'-( )-2'-deoxy- / J-thioguanylyl-(3 '-O—>5 '-O)-P-thiothymidylyl-(3 '-O—>5 '-O)-2'-deoxy-P-thioadenylyl-(3 '-O—>5 '-O)~ 2'-deoxy-5-methyl-P-thiocytidylyl-(3'-O— >5'-O)-2'-deoxy-5-methyl-P-thiocytidylyl-(3'-O^5'-O)-2'-deoxy-5-methyl- -thiocytidylyl-(3'-O—>5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiouridylyl-(3'-O— >5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3'-O^5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiouridylyl- PATENT 7328*158807PCT / NLM-1005-PC (3'-O^5'-O)-2'-O-(2-methoxyethyl)-5-methyl- -thiocytidylyl-(3'-<9— >5'-(9)-2'-(9-(2-methoxyethyl)-5-methylcytidine (SEQ ID NO:20), icosasodium salt.

[0206] B88. An oligomeric compound according to the general formula:

[0207]

[0208] B89. An oligomeric compound of any one of embodiments A1-B88, with the proviso that the oligomeric compound is not GalNAcoAmCmCmCATTGTGTAmCmCmCTmCTTmCmC (SEQ ID NO: 17), wherein each mC is a 5 -methylcytidine, GalNAc is a conjugate group containing multiple N-acetylgalactosamine moieties, and “o” is a phosphodiester moiety.

[0209] B90. The oligomeric compound of embodiment B89, wherein each internucleoside linkage of AmCmCmCATTGTGTAmCmCmCTmCTTmCmC (SEQ ID NO: 18) is a phosphorothioate intemucleoside linkage.

[0210] B91. An oligomeric compound of any one of embodiments A1-B90, with the proviso that the oligomeric compound oligonucleotide does not consist of the nucleobase sequence ACCCATTGTGTACCCTCTTCC (SEQ ID NO: 19).

[0211] B92. An oligomeric compound of any one of embodiments A1-B91, with the proviso that the oligomeric compound oligonucleotide does not consist of 21 linked nucleosides.

[0212] B93. An oligomeric compound of any one of embodiments A1-B92, comprising an oligonucleotide having one or more of the following features: (i) comprises one or more methyl uracil (mU) nucleosides; (ii) comprises one or more mU nucleosides and PATENT 7328*158807PCT / NLM-1005-PC one or more thymidine nucleosides; (iii) comprises two, or at least two, mU nucleosides, optionally within a terminal 3’ portion of the oligonucleotide; (iv) comprises two, or at least two, mU nucleosides and one or more thymidine nucleosides; and (v) consists of 20 linked nucleosides.

[0213] Cl. A composition, comprising an oligomeric compound of any one of embodiments A1-B93.

[0214] C2. The composition of embodiment Cl, wherein the oligomeric compound comprises at least one phosphorothioate internucleoside linkage, and the composition comprises a plurality of the oligomeric compound in a population of oligomeric compounds.

[0215] C3. The composition of embodiment C2, wherein the population is a chirally enriched population, wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate intemucleoside linkage having a particular stereochemical configuration.

[0216] C4. The composition of embodiment C3, wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a (Sp) configuration.

[0217] C5. The composition of embodiment C3, wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a (Rp) configuration.

[0218] C6. The composition of embodiment C3, wherein the population is enriched for modified oligonucleotides having a particular, independently selected stereochemical configuration at each phosphorothioate internucleoside linkage.

[0219] C7. The composition of embodiment C6, wherein the population is enriched for modified oligonucleotides having a (Sp) configuration at each phosphorothioate internucleoside linkage or for modified oligonucleotides having a (Rp) configuration at each phosphorothioate internucleoside linkage.

[0220] C8. The composition of embodiment C3, wherein the population is enriched for modified oligonucleotides having a (Rp) configuration at one particular phosphorothioate intemucleoside linkage and a (Sp) configuration at each of the remaining phosphorothioate internucleoside linkages. PATENT 7328*158807PCT / NLM-1005-PC C9. The composition of embodiment C3, wherein the population is enriched for modified oligonucleotides having at least 3 contiguous phosphorothioate internucleoside linkages in (Sp), (Sp) and (Rp) configurations, in the 5’ to 3’ direction. CIO. The composition of embodiment C2, wherein the phosphorothioate internucleoside linkages of the modified oligonucleotides in the population are stereorandom.

[0221] Cll. The composition of any one of embodiments Cl -CIO, wherein the modified oligonucleotides in the population is single stranded.

[0222] C12. A composition of any one of embodiments Cl-Cll, comprising an oligomeric duplex, wherein:

[0223] the oligomeric compound of any one of embodiments A1-B93 is a first oligomeric compound;

[0224] the oligomeric duplex comprises the first oligomeric compound and a second oligomeric compound; and

[0225] the second oligomeric compound comprises a second modified oligonucleotide.

[0226] C13. The composition of embodiment C12, wherein:

[0227] the second modified oligonucleotide consists of 12 to 50 linked nucleosides; and

[0228] the nucleobase sequence of the second modified oligonucleotide comprises a complementary region of at least 12 nucleobases that is at least 90% complementary to an equal length portion of the first modified oligonucleotide.

[0229] C14. The composition of embodiment Cll or C12, wherein the modified oligonucleotide of the first oligomeric compound and / or the second oligomeric compound comprises a 5’-stabilized phosphate group.

[0230] Cl 5. The composition of embodiment Cl 4, wherein the stabilized phosphate group comprises a cyclopropyl phosphonate or a vinyl phosphonate.

[0231] C16. The composition of any one of embodiments C12-C15, wherein at least one nucleoside of the second modified oligonucleotide comprises a modified sugar moiety.

[0232] Cl 7. The composition of embodiment Cl 6, wherein the modified sugar moiety of the second modified oligonucleotide comprises a bicyclic sugar moiety. PATENT 7328*158807PCT / NLM-1005-PC Cl 8. The composition of embodiment Cl 7, wherein the bicyclic sugar moiety comprises a 2’ -4’ bridge selected from -O-CH2- and -0-CH(CH3)-.

[0233] Cl 9. The composition of embodiment Cl 8, wherein the modified sugar moiety of the second modified oligonucleotide comprises a non-bicyclic modified sugar moiety. C20. The composition of embodiment Cl 9, wherein the non-bicyclic modified sugar moiety of the second modified oligonucleotide is a 2’-0Me sugar moiety or a 2’-F sugar moiety.

[0234] C21. The composition of any one of embodiments C12-C120, wherein at least one nucleoside of the second modified oligonucleotide comprises a sugar surrogate. C22. The composition of any one of embodiments C12-C21, wherein the second modified oligonucleotide comprises at least one modified internucleoside linkage. C23. The composition of embodiment C22, wherein the at least one modified internucleoside linkage of the second modified oligonucleotide is a phosphorothioate internucleoside linkage.

[0235] C24. The composition of any one of embodiments C12-C23, wherein the second modified oligonucleotide comprises at least one mesyl phosphoramidate (MsPA) linkage.

[0236] C25. The composition of embodiment C23, wherein each internucleoside linkage of the second modified oligonucleotide is independently selected from a phosphodiester internucleoside linkage and a phosphorothioate internucleoside linkage.

[0237] C26. The composition of any one of embodiments C12-C25, wherein the second modified oligonucleotide comprises at least one modified nucleobase.

[0238] C27. The composition of embodiment C26, wherein the at least one modified nucleobase is 5-methylcytosine.

[0239] C28. The composition of any one of embodiments C12-C27, wherein the second oligomeric compound comprises a conjugate group.

[0240] C29. The composition of embodiment C28, wherein the conjugate group comprises a conjugate moiety and a conjugate linker.

[0241] C30. The composition of embodiment C29, wherein the conjugate linker consists of a single bond.

[0242] C31. The composition of embodiment C29 or C30, wherein the conjugate linker is cleavable. PATENT 7328*158807PCT / NLM-1005-PC C32. The composition of any one of embodiments C29-C31, wherein the conjugate linker comprises 1-3 linker-nucleosides.

[0243] C33. The composition of any one of embodiments C29-C32, wherein the conjugate linker is a phosphodiester linker.

[0244] C34. The composition of any one of embodiments C28-C33, wherein the conjugate group is attached to the 5 ’-end of the second modified oligonucleotide.

[0245] C35. The composition of any one of embodiments C28-C33, wherein the conjugate group is attached to the 3 ’-end of the second modified oligonucleotide.

[0246] C36. The composition of any one of embodiments C28-C33, wherein the conjugate group is attached via the 2’ position of a ribosyl sugar moiety at an internal position of the second modified oligonucleotide.

[0247] C37. The composition of any one of embodiments C28-C36, wherein the conjugate group comprises a C22 alkyl, C20 alkyl, C16 alkyl, CIO alkyl, C21 alkyl, C19 alkyl, Cl 8 alkyl, C17 alkyl, Cl 5 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, Cll alkyl, C9 alkyl, C8 alkyl, C7 alkyl, C6 alkyl, C5 alkyl, C22 alkenyl, C20 alkenyl, C16 alkenyl, CIO alkenyl, C21 alkenyl, C19 alkenyl, Cl 8 alkenyl, C17 alkenyl, Cl 5 alkenyl, C14 alkenyl, C13 alkenyl, C12 alkenyl, Cl 1 alkenyl, C9 alkenyl, C8 alkenyl, C7 alkenyl, C6 alkenyl, or C5 alkenyl.

[0248] C38. The composition of any one of embodiments C28-C37, wherein the conjugate group comprises a cell -targeting moiety.

[0249] C39. The composition of any one of embodiments C28-C38, wherein the oligonucleotide compound is according to any one of embodiments B56-B82.

[0250] C40. The composition of any one of embodiments C12-C39, wherein the second modified oligonucleotide comprises a terminal group.

[0251] C41. The composition of embodiment C40, wherein the terminal group is an abasic sugar moiety.

[0252] C42. A composition, comprising an antisense agent, the antisense agent comprising or consisting of an antisense compound, wherein the antisense compound is the oligomeric compound of any one of embodiments A1-B93 or is in a composition of any one of embodiments Cl-Cll.

[0253] C43. A composition, comprising an antisense agent, wherein the antisense agent is the oligomeric duplex of any one of embodiments C12-C41.

[0254] C44. The composition of embodiment C42 or C43, wherein the antisense agent is: PATENT 7328*158807PCT / NLM-1005-PC (i) an RNase H agent capable of reducing the amount of SAA1 nucleic acid through the activation of RNase H; or

[0255] (ii) an RNAi agent capable of reducing the amount of SAA1 nucleic acid through the activation of RISC / Ago2.

[0256] C45. The composition of any one of embodiments C41-C44, wherein the antisense agent comprises a conjugate group, and wherein the conjugate group comprises a celltargeting moiety.

[0257] DI. A pharmaceutical composition comprising an oligomeric compound of any one of embodiments A1-B93 or a composition of any one of embodiments C1-C45, and a pharmaceutically acceptable diluent.

[0258] D2. The pharmaceutical composition of embodiment DI, wherein the pharmaceutically acceptable diluent is phosphate buffered saline (PBS) or artificial CSF (aCSF).

[0259] D3. The pharmaceutical composition of embodiment DI or D2, consisting essentially of the oligomeric compound of any one of embodiments A1-B93 or a composition of any one of embodiments C1-C45, and aCSF.

[0260] D4. The pharmaceutical composition of embodiment DI or D2, consisting essentially of the oligomeric compound of any one of embodiments A1-B93 or a composition of any one of embodiments C1-C45, and PBS.

[0261] El. A method for administering an oligomeric compound of any one of embodiments A1-B93, or a composition of any one of embodiments C1-C45, or pharmaceutical composition of any one of embodiments D1-D4, to a cell or tissue.

[0262] E2. A method for administering an oligomeric compound of any one of embodiments A1-B93, or a composition of any one of embodiments C1-C45, or pharmaceutical composition of any one of embodiments D1-D4, to a cell or tissue of a subject.

[0263] E3. The method of embodiment El or E2, wherein the oligomeric compound, composition or pharmaceutical composition is administered in vitro or ex vivo.

[0264] E4. The method of embodiment El or E2, wherein the oligomeric compound, composition or pharmaceutical composition is administered in vivo.

[0265] E5. The method of any one of embodiments E1-E4, wherein the oligomeric compound, composition or pharmaceutical composition is administered in an amount sufficient to reduce SAA1 expression. PATENT 7328*158807PCT / NLM-1005-PC E6. The method of embodiment E5, wherein the oligomeric compound, composition or pharmaceutical composition is administered in an amount sufficient to reduce SAA1 expression by 20% or more.

[0266] E7. A method for treating a SAA1 -associated medical condition, comprising administering an oligomeric compound of any one of embodiments A1-B93, or a composition of any one of embodiments C1-C45, or pharmaceutical composition of any one of embodiments D1-D4, to a cell or tissue of a subject in need thereof in an amount sufficient to treat the SAA1 -associated medical condition, wherein the SAA1-associated medical condition optionally is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia.

[0267] E8. The method of embodiment E7, wherein the oligomeric compound, composition or pharmaceutical composition is administered in an amount sufficient to reduce SAA1 expression.

[0268] E9. The method of embodiment E8, wherein the oligomeric compound, composition pharmaceutical composition is administered in an amount sufficient to reduce SAA1 expression by 20% or more.

[0269] E10. The method of any one of embodiments E7-E9, wherein at least one symptom or hallmark of an SAA1 -associated medical condition is ameliorated.

[0270] Ell. The method of embodiment E10, wherein the symptom or hallmark is one or more of proteinuria, nephrotic syndrome, low blood protein level, low blood albumin level, edema, high cholesterol, enlarged spleen, enlarged liver, enlarged thyroid, orthostatic hypotension, gastrointestinal atony, diarrhea and constipation.

[0271] E12. The method of any one of embodiments E7-E11, wherein administering the oligomeric compound, composition or pharmaceutical composition reduces or delays onset or progression of one or more symptoms or hallmarks of the SAA1 -associated medical condition.

[0272] E13. The method of any one of embodiments E1-E12, wherein the cell is or the tissue comprises a kidney cell or liver cell, or optionally a hepatocyte.

[0273] E14. The method of any one of embodiments E1-E13, wherein the cell is or the tissue comprises a human cell.

[0274] E15. The method of any of one of embodiments E2-E14, wherein the oligomeric compound, the composition or the pharmaceutical composition is administered by systemic administration, and optionally by subcutaneous administration. PATENT 7328*158807PCT / NLM-1005-PC E16. The method of embodiment E15, wherein the oligomeric compound, the composition, or the pharmaceutical composition is administered by subcutaneous injection.

[0275] E17. The method of any one of embodiments E2-E16, wherein the subject is human. El 8. The method of any one of embodiments El -El 7, wherein the cell or tissue comprises a SAA1 nucleic acid comprising a pathogenic genetic variant associated with a medical condition at a pathogenic genetic variation site and / or a pathogenic genetic variation position.

[0276] E19. The method of embodiment E18, wherein the pathogenic genetic variant is associated with a dominant negative effect, gain of function or toxic gain of function. E20. The method of embodiment E18 or E19, wherein the medical condition is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia.

[0277] E21. The method of any one of embodiments E18-E20, wherein the pathogenic genetic variation site comprises a pathogenic single nucleotide variation.

[0278] E22. The method of embodiment E21, wherein the pathogenic single nucleotide variation comprises a non-synonymous single nucleotide variation at a pathogenic genetic variation position.

[0279] E23. The method of any one of embodiments E18-E22, wherein a pathogenic genetic variant results in an amino acid substitution in a SAA1 protein selected from p. Leu3Pro, p. Val8Ala, p. Asp34Val, p. Ala70Val, p. Trp71Arg, p. Val75Ala, p. Asp93Glu, p. Gly90Asp, p. Asn101Asp and p. Lys121Arg.

[0280] E24. The method of any one of embodiments E18-E23, wherein the pathogenic genetic variant is in an exon of the SAA1 nucleic acid, optionally exon 3.

[0281] E25. The method of embodiment E24, wherein the pathogenic genetic variant results in a Asp34Val amino acid substitution, and optionally is c,101A> T.

[0282] E26. The method of any one of embodiments E1-E25, wherein the cell, tissue and / or subject comprise an elevated SAA1 level.

[0283] E27. The method of embodiment E26, wherein the elevated SAA1 level is an elevated level of a SAA1 nucleic acid, or SAA1 protein or SAA1 nucleic acid and SAA1 protein.

[0284] E28. The method of embodiment E26 or E27, wherein the elevated SAA1 level is in blood, or optionally in serum, or optionally in plasma. PATENT 7328*158807PCT / NLM-1005-PC Fl. The method of any one of embodiments E18-E25, comprising determining presence or absence of the pathogenic genetic variant; and if the pathogenic genetic variant is present, administering the oligomeric compound, composition or pharmaceutical composition.

[0285] F2. The method of embodiment Fl, comprising:

[0286] genotyping a genetic variation position and / or a genetic variation site of the SAA1 nucleic acid, thereby providing a SAA1 nucleic acid genotype; and determining presence or absence of the pathogenic genetic variant according to the genotype.

[0287] F3. The method of embodiment Fl or F2, wherein the SAA1 nucleic acid is from a cell or tissue.

[0288] F4. The method of any one of embodiments F1-F3, wherein the SAA1 nucleic acid is from a subject.

[0289] F5. The method of embodiment F3 or F4, wherein the cell, tissue or subject is a human cell, human tissue or human subject.

[0290] F6. The method of any one of embodiments F1-F5, wherein the SAA1 nucleic acid is from a subject having a medical condition.

[0291] F7. The method of embodiment F6, wherein the medical condition is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia.

[0292] F8. The method of any one of embodiments F1-F7, wherein determining the presence or absence of the genetic variant, and optionally the genotyping, comprises sequencing the SAA1 nucleic acid.

[0293] F9. The method of embodiment F8, wherein the sequencing comprises short-read sequencing, or long-read sequencing, or short-read sequencing and long-read sequencing.

[0294] F10. The method of any one of embodiments F1-F9, wherein a pathogenic genetic variant results in an amino acid substitution in a SAA1 protein selected from p. Leu3Pro, p. Val8Ala, p. Asp34Val, p. Ala70Val, p. Trp71Arg, p. Val75Ala, p. Asp93Glu, p. Gly90Asp, p. Asn101Asp and p. Lys121Arg.

[0295] Fll. The method of any one of embodiments F1-F10, wherein the pathogenic genetic variant is in an exon of the SAA1 nucleic acid, optionally exon 3. PATENT 7328*158807PCT / NLM-1005-PC F12. The method of embodiment Fl 1, wherein the pathogenic genetic variant results in a Asp34Val amino acid substitution, and optionally is c,101A> T.

[0296] F13. The method of any one of embodiments F1-F12, comprising administering an oligomeric compound that targets a SAA1 nucleic acid if presence of the pathogenic genetic variant genotype is determined.

[0297] F14. The method of any one of embodiments E1-E28 and F1-F13, comprising determining presence, absence and / or amount of a SAA1 level, which optionally is an elevated SAA1 level, wherein the SAA1 level optionally is a level of a SAA1 nucleic acid and / or SAA1 protein.

[0298] F15. The method of embodiment Fl 4, wherein the SAA1 level, optionally the level of the SAA1 nucleic acid or SAA1 protein, is relative to a control SAA1 level.

[0299] F16. The method of embodiment Fl 5, wherein the control SAA1 level is in a cell or cells in which SAA1 nucleic acid does not contain a pathogenic genetic variant. F17. The method of any one of embodiments F14-F16, comprising administering an oligomeric compound targeted to a SAA1 nucleic acid if presence of an elevated SAA1 level, optionally an elevated level of a SAA1 nucleic acid and / or SAA1 protein, is determined.

[0300] F18. The method of any one of embodiments F14-F17, wherein presence, absence and / or amount of a SAA1 level, is determined for a sample, a cell, cells, a tissue and / or a subject.

[0301] F19. The method of any one of embodiments F14-F18, wherein the SAA1 level is in blood, or optionally in serum, or optionally in plasma.

[0302] F20. The method of any one of embodiments F14-F19, wherein the oligomeric compound targeted to a SAA1 nucleic acid is administered to a cell, cells, a tissue and / or a subject.

[0303] F21. The method of any one of embodiments F14-F20, wherein the SAA1 nucleic acid is SAA1 RNA, and optionally the RNA is pre-mRNA and / or mRNA.

[0304] F22. The method of any one of embodiments F1-F21, wherein the oligomeric compound is of any one of embodiments A1-B93, the composition is of any one of embodiments C1-C45, or the pharmaceutical composition is of any one of embodiments D1-D4. PATENT 7328*158807PCT / NLM-1005-PC Gl. An oligomeric compound of any one of embodiments A1-B93, a composition of any one of embodiments C1-C45, or a pharmaceutical composition of any one of embodiments D1-D4, for reducing SAA1 expression.

[0305] G2. An oligomeric compound of any one of embodiments A1-B93, a composition of any one of embodiments C1-C45, or a pharmaceutical composition of any one of embodiments D1-D4, for treating a medical condition.

[0306] G3. Use of an oligomeric compound of any one of embodiments A1-B93, a composition of any one of embodiments C1-C45, or a pharmaceutical composition of any one of embodiments D1-D4, for reducing SAA1 expression.

[0307] G4. Use of an oligomeric compound of any one of embodiments A1-B93, a composition of any one of embodiments C1-C45, or a pharmaceutical composition of any one of embodiments D1-D4, for treating a medical condition.

[0308] G5. Use of oligomeric compound of any one of embodiments A1-B93, a composition of any one of embodiments C1-C45, or a pharmaceutical composition of any one of embodiments D1-D4, for the manufacture of a medicament for reducing SAA1 expression.

[0309] G6. Use of oligomeric compound of any one of embodiments A1-B93, a composition of any one of embodiments C1-C45, or a pharmaceutical composition of any one of embodiments D1-D4, for the manufacture of a medicament for treating a medical condition.

[0310] G7. The oligomeric compound, composition, pharmaceutical composition or use of any one of embodiments G2, G4 or G6, wherein the medical condition is a SAA1-associated medical condition, and optionally is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia.

[0311] G8. The oligomeric compound, composition, pharmaceutical composition or use of any one of embodiments G1-G7, for administration according to any one of embodiments E1-E28 or F1-F22.

[0312] Chimeric Oligonucleotides

[0313] In certain embodiments, provided are oligomeric compounds comprising oligonucleotides that consist of linked nucleosides. Oligonucleotides can be unmodified oligonucleotides (RNA or DNA) or can be modified oligonucleotides. Modified oligonucleotides comprise at least one modification relative to unmodified RNA or DNA. Modified oligonucleotides comprise at least one modified nucleoside PATENT 7328*158807PCT / NLM-1005-PC (comprising a modified sugar moiety and / or a modified nucleobase) and / or at least one modified internucleoside linkage. Certain modified nucleosides and modified internucleoside linkages suitable for use in modified oligonucleotides are described.

[0314] Nucleosides

[0315]

[0316] Modified nucleosides comprise a modified sugar moiety or a modified nucleobase or both a modified sugar moiety and a modified nucleobase. In certain embodiments, modified nucleosides comprising the fol lowing modified sugar moieties and / or the following modified nucleobases can be incorporated into antisense oligonucleotides.

[0317] Exemplary Sugar Moieties

[0318] In certain embodiments, modified sugar moieties are non-bicyclic modified sugar moieties. In certain embodiments, modified sugar moieties are bicyclic or tricyclic sugar moieties. In certain embodiments, modified sugar moieties are sugar surrogates. Such sugar surrogates may comprise one or more substitutions corresponding to those of other types of modified sugar moieties.

[0319] In certain embodiments, modified sugar moieties are non-bicyclic modified furanosyl sugar moieties comprising one or more acyclic substituent, including, but not limited to, substituents at the 2’, 3’, 4’, and / or 5’ positions. In certain embodiments, the furanosyl sugar moiety is a ribosyl sugar moiety. In certain embodiments, one or more acyclic substituent of non-bicyclic modified sugar moieties is branched.

[0320] In certain embodiments, non-bicyclic modified sugar moieties comprise a substituent group at the deposition, referred to as 2 ’-substituted sugar moieties.

[0321] Examples of substituent groups suitable for the 2’ -position of modified sugar moieties include but are not limited to: -F, -OCH? (“OMe” or “O-methyl” or “'methoxy”), and -OCH2CH2OCH3 (“MOE’’ or “O-methoxy ethyl”). In certain embodiments, 2’-substituent groups are selected from among: halo, allyl, amino, azido, SH, CN, OCN, CF3, OCF3, O-C1-C10 alkoxy, O-C1-C10 substituted alkoxy, O-Ct-Cio alkyl, O-Ci-Cio substituted alkyd, S-alkyl, ty Rir:)-alkyl, O-alkenyl, S-alkenyl, N(R.ir:)-alkenyl, O-alkynyl, S-alkynyl, N(Rm)-alkynyl, O-alkylenyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryi, O-aralkyl, (XCHtysSCHs, 0(CH ) 0N(R )(Rty or 0CH2C(=0)-N(Rra)(R:i), where each m and Rn is, independently, hydrogen, an amino protecting group, or substituted or unsubstitutedCi-Cio alkyl, -O(CH2)2ON(CH3)2 (“DMAOE”), 2’- PATENT 7328*158807PCT / NLM-1005-PC O(CH2)2O(CH2)2N(CH3)2(“DMAEOE”), and the 2’ -substituent groups described in Cook et al., U. S. 6,531,584; Cook el al., U. S. 5.859.221: and Cook et al., U. S.

[0322] 6,005,087. Certain embodiments of these 2'-substituent groups can be further substituted with one or more substituent groups independently selected from among: hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro (NO?.), thiol, thioalkoxy, thioalkyl, halogen, alkyl, and, alkenyl and alkynyl.

[0323] In certain embodiments, a 2’-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2’-substituent group selected from: F, Ni b. N3, OCFy OCH.r O(CH2)3NH2, Cl bCH Ci L. OCH2C H CM2.

[0324] OCH2CH2OCH3 • • MOE ), O(C; H2)?. SCH 3, O(C 1H 2 )?. ON( R )(IU ), O(CH2)2O(CH2)2 (CH3)2, and N-substituted acetamide (OCH2C(===O)-N(Rm)(R1)), where each

[0325]

[0326] and IU is, independently, hydrogen, an amino protecting group, or substituted or unsubstituted Ci~Cio alkyl.

[0327] In certain embodiments, a 2 ’-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2’ -sub tituent group selected from: F, OCF3, (H i b. OCH2C I bOCI is. O(CH2)2SCH3, O(CH2)2ON(CI b )2, O(CH2)2O(CH?)2N(CH3)2, O(CH2)2O (CHS)2(“DMAOE”), O(CH2)2O(CH2)2N(CH3)2(“DM AEOE”), and OCH2G OMM i bCl b (“NMA”). In certain embodiments, a 2’ -substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2 ’-substituent group selected from: F, OCH3, OCH? CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(CH3)2, O(CH2)2O(CH2)2N(CH3)2, and OCH2C(===O)-N(H)CI-b (“NMA”).

[0328] In certain embodiments, a 2 ’-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2’ -sub tituent group selected from: F, OCH3, and OCH2CH2OCH3.

[0329] In certain embodiments, modified furanosyl sugar moieties and nucleosides incorporating such modified furanosyl sugar moieties are further defined by isomeric configuration. For example, a 2’-deoxyfuranosyi sugar moiety can be in seven isomeric configurations other than the naturally occurring beta-D-deoxyribosyl configuration. Such modified sugar moieties are described in, for example, WO2019 / 157531. A 2’-modified sugar moiety has an additional stereocenter at the deposition relative to a 2 ’-deoxy furanosyl sugar moiety; therefore, such sugar moieties have a total of sixteen possible isomeric configurations. 2’ -modified sugar moieties PATENT 7328*158807PCT / NLM-1005-PC described herein are in the beta-D-ribosyl isomeric configuration unless otherwise specified.

[0330] In certain embodiments, non-bicyclic modified sugar moieties comprise a substituent group at the 4’-position. Examples of' substituent groups suitable for the 4’-position of modified sugar moieties include but are not limited to alkoxy (for example, methoxy), alkyl, and those described in Manoharan et al., W02015 / 106128. In certain embodiments, non-bicyclic modified sugar moieties comprise a substituent group at the 3 ’-position. Examples of substituent groups suitable for the 3 ’-position of modified sugar moieties include, but are not limited to, alkoxy (for example, methoxy) and alkyl (for example, methyl, ethyl).

[0331] In certain embodiments, non-bicyclic modified sugar moieties comprise a substituent group at the 5 ’-position. Examples of substituent groups suitable for the 5 ’-position of modified sugar moieties include, but are not limited to, vinyl, alkoxy (for example, methoxy), and alkyl (e g., methyl (R. or S), ethyl).

[0332] In certain embodiments, non-bicyclic modified sugar moieties comprise more than one non-bridging sugar substituent, for example, 2!-F-5!-methyl sugar moieties and the modified sugar moieties and modified nucleosides described in Migawa et al., W02008 / 101157 and Rajeev et al., US2013 / 0203836).

[0333] In naturally occurring (unmodified) nucleic acids, sugars are linked to one another 3’ to 5’. In certain embodiments, oli onucleotides include one or more nucleoside or sugar moiety linked at an alternative position, for example at the 2’ position or inverted 5’ to 3’. For example, where the linkage is at the 2’ position, the 2’-substituent groups may instead be at the 3 ’-position.

[0334] Certain modified sugar moieties comprise a substituent that bridges two atoms of the furanosy 1 ring to form a second ring, resulting in a bicyclic sugar moiety. In certain embodiments, the bicyclic sugar moiety comprises a bridge between the 4‘ and the 2‘ furanose ring atoms. Examples of such 4’ to 2’ bridging sugar substituents include, but are not limited to: 4'-CH?.-2', 4-(CH2)?.-2', 4'-(CH2)3-2', 4-CH2-O-2' (“LNA”), 4-CH2-S-2’, 4’-(CH2)2-O-2’ (“ENA”), 4’-CH(CH3)-O-2' (referred to as “constrained ethyl” or “cEt”), 4’-CH2-O-CH2-2’, 4’-CH?.-N(R)-2.’, 4’-CH(CH?. OCH3)-0-2' (“constrained MOE” or “cMOE”) and analogs thereof (see, for example, Seth et al., U. S. 7,399,845, Bhat et al., U. S. 7,569,686, Swayze et al., U. S. 7,741,457, and Swayze et al., U. S. 8,022,193), 4'-C(CH<)(CH3)-O~2’ and analogs thereof (see, for PATENT 7328*158807PCT / NLM-1005-PC example, Seth et al., U. S. 8,278,283), 4'-CH2-N(OCH?)"2’ and analogs thereof (see, for example, Prakash et al., U. S. 8,278,425), 4'-CH2-O-N(CFl3)-2' (see, for example, Allerson et al., U. S. 7,696,345 and Allerson et al., U. S. 8,124,745), 4-CH?.-C(H)(CH3)-2' (see, for example, Zhou, et al., J. Org. Chem.,2009, 74, 118-134), 4'-CH2”C(==CH?.)-2' and analogs thereof (see for example,, Seth et al., U. S. 8,278,426), 4’-C(RaRb)-N(R)-O-2’, 4’-C(RJU)-O-N(R)-2’, 4'-CH2-O-N(R)-2’, and 4'-CH2-N(R)-0-2', where each R, Ra, and Rb, is, independently, hydrogen, a protecting group, or Ci-Cn alkyl (see, for example Imanishi et al., U. S. 7,427,672),

[0335] In certain embodiments, such 4’ to 2’ bridges independently comprise from 1 to 4 linked groups independently selected from: -[C(R )(Rb)];i-, -[CnUJtRbtJii-O-, -C(R0===C(Rr)-, -C(Ra):::N~, -Ct \R,p. 41 Ok -C( S ) - -O-, ~Si( a, -St Uy.-, and -N(Ra)-; where: x is 0, 1, or 2; n is 1, 2, 3, or 4; each Ra and Rb is, independently, hydrogen, a protecting group, hydroxyl, Ci-Cis alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen. Oh, NJ1J2, SJi, Ns, COOJi, acyl (C(=0)-H), substituted acyl, CN, sulfonyl ( S{ O) 2- J 1 ), or sulfoxyl (S(:

[0336]

[0337] ); and each Ji and J2is, independently, hydrogen, C1-C12 alkyl, substituted C1-C12 alkyl, CS-CJ?. alkenyl, substituted C2-CJ2 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, acyl (C(=0)-H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, Ci-C aminoalkyl, substituted C1-C12 aminoalkyl, or a protecting group.

[0338] Additional bicyclic sugar moietles are known in the art, see, for example: Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443, Albaek et al., J. Org. Chem., 2006, 71, 7731-7740, Singh et al., Chem. Common,, 1998, 4. 455-456:

[0339] Koshkin et al.. Tetrahedron, 1998, 54, 3607-3630: Wahlestedt et al.. Proc. Natl. Acad. Sci. U. S. A., 2000, 97, 5633-5638; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivasiava et al., J. Am. Chem. Soc., 2007, 129, 8362-8379; Elayadi et al., Curr. Opinion Inverts. Drugs, 2001, 2, 558-561; Braasch et al., Chem. Biol., 2001, 8, 1-7; Orunr et al., Curr.

[0340] Opinion Mol. Then, 2001, 3, 239-243; VVengel etal., U. S. 7,053,207, Imanishi et al., U. S. 6,268,490, Imanishi et al. U. S. 6,770,748, Imanishi et al., U. S. RE44,779; PATENT 7328*158807PCT / NLM-1005-PC Wengel et al., U. S. 6,794,499, Wengel et al., U. S. 6,670,461; Wengel et al.. U. S. 7,034,133, Wengel et al., U. S. 8,080,644; Wengel et al„ U. S. 8,034,909; Wengel et al., U. S. 8,153,365; Wengel et al., U. S. 7,572,582; and Ramasamy et al., U. S.

[0341] 6,525,191, Torsten et al., WO 2004 / 106356, Wengel et al., WO 1999 / 014226; Set et al. WO 2007 / 134181; Seth et al., U. S. 7,547,684; Seth et ah, U. S. 7,666,854; Seth et al., U. S. 8,088,746; Seth et al., U. S. 7,750,131; Seth et al., U. S. 8,030,467; Seth et al., U. S. 8,268,980, Seth et al., U. S. 8,546,556, Seth et al., U. S. 8,530,640; Migawa et al., U. S. 9,012,421; Seth et al., U. S. 8,501,805; Allerson et al., US2008 / 0039618; and Migawa et al., US2015 / 0191727.

[0342] In certain embodiments, bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by isomeric configuration. For example, an LNA nucleoside (described herein) can be in the alpha-L configuration or in the beta-D configuration.

[0343]

[0344] LNA (beta-D-configurati on) alpha-L -UNA (alpha-L-configurati on)

[0345] bridge = 4'-CH2-O-2’ bridge = 4‘-CH2-O-2'

[0346] Alpha-L-methyleneoxy (4’-CH2-O-2!) or alpha-L -LNA bicyclic nucleosides have been incorporated into oligonucleotides that showed antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372). The addition of locked nucleic acids to siRNAs has been shown to increase siRNA stability in serum, and to reduce off-target effects (El men, J. et al., (2005) Nucleic Acids Research 33(l):439-447; Mook, OR. et al., (2007) Mai Cane Ther 6(3):833-843; Grunweller, A. et al., (2003) Nucleic Acids Research 31(12):3185-3193). Herein, general descriptions of bicyclic nucleosides include both isomeric configurations. When the positions of specific bicyclic nucleosides (for example, LNA or cEt) are identified in embodiments herein, they are in the beta-D confi uration, unless otherwise specified.

[0347] In certain embodiments, modified sugar moieties comprise one or more non-bridging sugar substituent and one or more bridging sugar substituent (for example, o’substituted and 4’-2’ bridged sugars).

[0348] In certain embodiments, modified sugar moieties are sugar surrogates. In certain such embodiments, the oxygen atom of the sugar moiety is replaced, for PATENT 7328*158807PCT / NLM-1005-PC example, with a sulfur, carbon or nitrogen atom. In certain such embodiments, such modified sugar moieties also comprise bridging and / or non-bridging substituents as described herein. For example, certain sugar surrogates comprise a 4’-sulfur atom and a substitution at the 21~position (see, for example, Bhat et al., U. S. 7,875,733 and Bhat. et al., U. S. 7,939,677) and / or the 5’ position.

[0349] In certain embodiments, sugar surrogates comprise rings having other than 5 atoms. For example, in certain embodiments, a sugar surrogate comprises a sixmembered tetrahydropyran (“THP”). Such tetrahydropyrans can be further modified or substituted. Nucleosides comprising such modified tetrahydropyrans include but are not limited to hexitol nucleic acid ("‘HNA”), anitol nucleic acid (“ANA”), mannitol nucleic acid (“MN A”) (see, for example, Leumann, CJ. Bioorg. & Med. Chem. 2002, 10, 841-854), fluoro HNA:

[0350]

[0351] F-HNA

[0352] (“F-HNA”, see for example, Swayze et al., U. S. 8,088,904; Swayze et al., U. S.

[0353] 8,440,803; Swayze et ah, U. S. 8,796,437, and Swayze et ah, U. S. 9,005,906, F-HNA can also be referred to as a F-THP or 3 ~fluoro tetrahydropyran), and nucleosides comprising additional modified THP compounds having the formula:

[0354]

[0355] where, independently, for each of the modified THP nucleoside:

[0356] Bx is a nucleobase moiety;

[0357] Ts and 4 each, independently, is an internucleoside linking group linking the modified THP nucleoside to the remainder of an oligonucleotide or one of lb and T4 is an internucleoside linking group linking the modified THP nucleoside to the remainder of an oligonucleotide and the other of Ts and Tr is hydrogen, a hydroxyl protecting group, a linked conjugate group, or a 5‘ or 3’-terminal group; qj, qs, qs, qy qs, qs and q? are each, independently, hydrogen, Ci-C-6 alkyl, substituted Ci-Cs alkyl, PATENT 7328*158807PCT / NLM-1005-PC C / j-Cf, alkenyl, substituted Cz-C alkenyl, C2-C6 alkynyl, or substituted Cz-C-6 alkynyl; and each of Ri and R2 is independently selected from among hydrogen, halogen, substituted or unsubstituted alkoxy, NJ1J2, SJi, Ng OC(=X)Ji, OC(=X)NJ 1J2, XUC(::X)NJ: J 2, and CN, where Xis O, S or NJi and each Ji, h, and h is, independently, hydrogen or Ci-Cs alkyl.

[0358] In certain embodiments, modified THP nucleosides are provided where qn q2, qs, qa, q.s, qs and q? each is hydrogen. In certain embodiments, at least one of qi, qz, qy q4, q.y qs and q? is other than hydrogen. In certain embodiments, at least one of qi, qs, qs, qy qs, q& and q? is methyl. In certain embodiments, modified THP nucleosides are provided where one of Ri and R2 i fluorine. In certain embodiments, Ri is fluorine and R? is hydrogen, in certain embodiments, Rj is methoxy and R2 is hydrogen, and in certain embodiments, Ri is methoxyethoxy and R2 is hydrogen. In certain embodiments, sugar surrogates comprise rings having more than 5 atoms and more than one heteroatom. For example, nucleosides compri ing morpholino sugar moieties and their use in oligonucleotides have been reported (see, for example, Braasch et al., Biochemistry, 2002, 41, 4503-4510 and Summerton et ai., U. S.

[0359] 5,698,685; Summerton et al., U. S. 5,166,315; Summerton et al., U. S. 5,185,444; and Summerton et al., U. S. 5,034,506). As used here, the term “'morpholino’'' means a sugar surrogate that can be in a nucleoside having the followdng structure, w'here Bx is a nucleobase moiety:

[0360]

[0361] In certain embodiments, morpholinos can be modified, for example by adding or altering various substituent groups from the above morpholino structure. Such sugar surrogates are referred to herein as “modified morpholinos.”

[0362] In certain embodiments, sugar surrogates comprise acyclic moieties. Examples of nucleosides and oligonucleotides comprising such acyclic sugar surrogates include, but are not limited to, peptide nucleic acid (“PNA”), acyclic butyl nucleic acid (see, for example, Kumar et al., Org. Biomol. Chern., 2013, 11, 5853-5865), and nucleosides and oligonucleotides described in Manoliaran et al., WO2011 / 133876. In certain embodiments, sugar surrogates comprise acyclic moieties. Examples of PATENT 7328*158807PCT / NLM-1005-PC nucleosides and oligonucleotides comprising such acyclic sugar surrogates include, but are not limited to, peptide nucleic acid (“PNA”), acyclic butyl nucleic acid (see, for example, Kumar et al., Org. Biomol. Chem., 2013, 11, 5853-5865), and nucleosides and oligonucleotides described in Manoharan et al., US2013 / 130378. Representative U. S. patents that teach the preparation of PNA compounds include, but are not limited to, U. S. Patent Nos. 5,539,082; 5,714,331; and 5,719,262. Additional PNA compounds suitable for use in the oligonucleotides of the invention are described in, for example, in Nielsen et al., Science, 1991, 254, 1497-1500.

[0363] In certain embodiments, sugar surrogates are the “unlocked” sugar structure of UNA (unlocked nucleic acid) nucleosides. UNA is an unlocked acyclic nucleic acid, where any of the bonds of the sugar has been removed, forming an unlocked sugar surrogate. Representative U. S. publications that teach the preparation of UNA include, but are not limited to, US Patent No. 8,314,227; and US Patent Publication Nos. 2013 / 0096289; 2013 / 0011922; and 2011 / 0313020.

[0364] In certain embodiments, a sugar surrogate is a glycerol as found in GNA (glycol nucleic acid) nucleosides as depicted below:

[0365] (S)-GNA

[0366]

[0367] where Bx represents a nucleobase moiety.

[0368] Many other bicyclic and tricyclic sugar and sugar surrogates are known in the art that can be used in modified nucleosides.

[0369] Certain Modified Niicleabases

[0370] In certain embodiments, modified oligonucleotides comprise one or more nucleoside comprising an unmodified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more nucleoside comprising a modified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more nucleoside that does not comprise a nucleobase, referred to as an abasic nucleoside. In certain embodiments, modified oligonucleotides comprise one or more inosine nucleosides (i.e., nucleosides comprising a hypoxanthine nucleobase). PATENT 7328*158807PCT / NLM-1005-PC In certain embodiments, modified nucleobases are selected from: 5-substituted pyrimidines, 6-azapyrimidines, alkyl or alkynyl substituted pyrimidines, alkyl substituted purines, and N-2, N-6 and 0-6 substituted purines. In certain embodiments, modified nucleobases are selected from: 5-methylcytosine, 2-aminopropy I adenine, 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2~propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (-C^C-CIE) uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8 -substituted purines, 5-halo, particularly 5-bromo, 5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine, 7-m ethyl adenine, 2-F-adenine. 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3 -deazaguanine, 3 -deazaadenine, 6-N-benzoyladenine, 2-N-isobutyryl guanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5 -methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. Further modified nucleobases include tricyclic pyrimidines, such as 1,3-diazaphenoxazine-2-one, l,3-diazaphenothiazine-2-one and 9-(2-aminoethoxy)-l,3-di zaphenoxazine-2-one (G-clamp). Modified nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-ami nopyridine and 2-pyridone. Further nucleobases include those disclosed in Merigan et al., U. S. 3,687,808, those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, Kroscbwitz, J. I., Ed., John Wiley & Sons, 1990, 858-859; Engl i sc h et al., Angewandte Chemi e, International Edition, 1991, 30, 613; Sanghvi, Y. S., Chapter 15, Antisense Research and Applications, Crooke, S. T. and Lebleu, B., Eds., CRC Press, 1993, 273-288; and those disclosed in Chapters 6 and 5, Antisense Drug Technology, Crooke S. T., Ed., CRC Press, 2008, 163-166 and 442-443.

[0371] Publications that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include without limitation, Manoharan et al., US 2003 / 0158403; Manoharan et al., US 2003 / 0175906; Dinh et al., U. S. 4,845,205; Spielvogel et al., U. S. 5,130,302; Rogers et al., U. S. 5,134,066;

[0372] Bischofberger et al., U. S. 5,175,273; Urdea et al., U. S. 5,367,066; Benner et al., U. S.

[0373] 5,432,272; Matteucci et al., U. S. 5,434,257; Grneiner et al., U. S. 5,457,187; Cook et PATENT 7328*158807PCT / NLM-1005-PC al., U. S. 5,459,255; Froehler et al., U. S. 5,484,908; Matteucci et al., U. S. 5,502,177; Hawkins et al., U. S. 5,525,711, Flaralambidis et al., U. S. 5,552,540; Cook et al., U. S.

[0374] 5,587,469; Froehler et al., U. S. 5,594,121; Switzer et al., U. S. 5,596,091; Cook et al., U. S. 5,614,617; Froehler et al., U. S. 5,645,985; Cook et al., U. S. 5,681,941; Cook et al., U. S. 5,811,534; Cook et al., U. S. 5,750,692; Cook et al., U. S. 5,948,903; Cook et al., U. S. 5,587,470; Cook et al., U. S. 5,457,191; Matteucci et al., U. S. 5,763,588; Froehler et al., U. S. 5,830,653, Cook et al., U. S. 5,808,027; Cook et al., U. S.

[0375] 6,166,199; and Matteucci et al., U. S. 6,005,096.

[0376] IMmipmx Modified interne J ecsUe tankages

[0377] The naturally occurring internucleoside linkage of RNA and DNA is a 3' to 5’ phosphodiester linkage. In certain embodiments, nucleosides of modified oligonucleotides can be linked together using one or more modified internucleoside linkages. The two main classes of intemudeoside linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus-containing internucleoside linkages include, but are not limited to, phosphodiesters, which contain a phosphodiester bond (" P O' ) (also referred to as unmodified or naturally occurring linkages), phosphotriesters, methylphosphonates, phosphorarnidates, phosphorothi oates (“P^S”), and phosphorodithi oates (“HS-P::::S”). Representative non-phosphorus containing internucleoside linking groups include, but are not limited to, methylenemethylimino (-CHr-NtCHtQ-O-CH;-), thiodi ester, thionocarbamate (-O-C(=O)('NH)-S“); siloxane (-O-SiH?.-O-); and N, N‘-dimetliylhydrazine (-CH2-N(CHs)-N(CHs)-). Modified internucleoside linkages, compared to naturally occurring phosphodi ester internucleoside linkages, can be used to alter, typically increase, nuclease resistance of the oligonucleotide. In certain embodiments, internucleoside linkages having a chiral atom can be prepared as a racemic mixture, or as separate enantiomers. Methods of preparation of phosphorous-containing and non-phosphorous-containing internucleoside linkages are well known to those skilled in the art.

[0378] In certain embodiments, a modified internucleoside linkage is any of those described in WO 2021 / 030778. In certain embodiments, a modified internucleoside linkage comprises the formula: PATENT 7328*158807PCT / NLM-1005-PC

[0379]

[0380] where independently for each internucleoside linking group of the modified oligonucleotide:

[0381] X is selected from 0 or S;

[0382] R1 is selected from hydrogen, C1-C6 alkyl, and substituted C1-C6 alkyl; and

[0383] T is selected from SO2R2, C(=O)R3, and P(=O)R4R5, where:

[0384] R2 is selected from an aryl, a substituted aryl, a heterocycle, a substituted heterocycle, an aromatic heterocycle, a substituted aromatic heterocycle, a diazole, a substituted diazole, a C1-C6 alkoxy, C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, substituted C1-C6 alkyl, substituted C1-C6 alkenyl substituted C1-C6 alkynyl, and a conjugate group; R3 is selected from an aryl, a substituted aryl, CH3, N(CH3)2, OCH3 and a conjugate group;

[0385] R4 is selected from OCH3, OH, C1-C6 alkyl, substituted C1-C6 alkyd and a conjugate group; and

[0386] R5 is selected from OCH3, OH, C1-C6 alkyl, and substituted C1-C6 alkyl.

[0387] In certain embodiments, a modified intemucleoside linkage comprises a mesyl phosphoramidate linking group having a formula:

[0388]

[0389] In certain embodiments, a mesyl phosphoramidate internucleoside linkage may comprise a chiral center. In certain embodiments, modified oligonucleotides comprising (Rp) and / or (Sp) mesyl phosphoramidates comprise one or more of the following formulas, respectively, where “B” indicates a nucleobase; PATENT 7328*158807PCT / NLM-1005-PC

[0390]

[0391] In certain embodiments, a modified intemucleoside linkage comprises a busy! phosphorami date linking group having a formula:

[0392]

[0393] A busyl phosphora idate intemucleoside linkage may comprise a chiral center. In certain embodiments, a modified oligonucleotide can include (Rp) and / or (Sp) busyl phosphorami dates having structures corresponding to those shown for (Rp) or (Sp) mesyl phosphoramidates, respectively.

[0394] Representative internucleoside linkages having a chiral center include but are not limited to alkylphosphonates, phosphoramidates, and phosphor othi oates. Modified oligonucleotides comprising internucleoside linkages having a chiral center can be prepared as populations of modified oligonucleotides comprising stereo-random internucleoside linkages, or as populations of modified oligonucleotides comprising phosphorothioate linkages in particular stereochemical configurations. In certain embodiments, populations of modified oligonucleotides comprise phosphorothioate internucleoside linkages where all of the phosphorothioate internucleoside linkages arc stereo-random. Such modified oligonucleotides can be generated using synthetic methods that result in random selection of the stereochemical configuration of each phosphorothioate linkage. Nonetheless, each individual phosphorothioate of each individual oligonucleotide molecule has a defined stereo-configuration. In certain embodiments, populations of modified oligonucleotides are enriched for modified oligonucleotides comprising one or more particular phosphorothioate internucleoside linkages in a particular, independently selected stereochemical configuration. PATENT 7328*158807PCT / NLM-1005-PC In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 65% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 70% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 80% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 90% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 99% of the molecules in the population. Such chirally enriched populations of modified oligonucleotides can be generated using synthetic methods known in the art, for example, methods described in Oka et al., JACS 125, 8307 (2003), Wan et al. Nuc. Acid. Res. 42, 13456 (2014), and WO 2017 / 015555. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one indicated phosphorothioate in the (Sp) configuration. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one phosphorothioate in the (Rp) configuration. In certain embodiments, modified oligonucleotides comprising (Rp) and / or (Sp) phosphorothioates comprise one or more of the following formulas, respectively, where “B” indicates a nucleobase:

[0395]

[0396] Unless otherwise indicated, chiral internucleoside linkages of modified oligonucleotides described herein can be stereo-random or in a particular stereoch emi cal c onfi gurati on.

[0397] Neutral internucleoside linkages include, without limitation, phosphotri esters, alkylphosphonates, methylphosphonates, MMI (3'-CH2-N(CH3)-O-5'), amide-3 (3'- PATENT 7328*158807PCT / NLM-1005-PC CH2-C(=O)-N(H)-5'), amide-4 (3'-CH2-N(H)-C(=O)-5'), formacetal (3'-O-CH2-O-5'), methoxypropyl (MOP), and thioformacetal (3'-S-CH2-O-5'). Further neutral internucleoside linkages include nonionic linkages comprising siloxane (di lkylsiloxane), carboxylate ester, carboxamide, sulfide, sulfonate ester and amides (See for example: Carbohydrate Modifications in Antisense Research', Y. S. Sanghvi and P. D. Cook, Eds., ACS Symposium Series 580; Chapters 3 and 4, 40-65). Further neutral internucleoside linkages include nonionic linkages comprising mixed N, O, S and CH2 component parts. In certain embodiments, modified oligonucleotides comprise one or more inverted nucleoside, as shown below:

[0398]

[0399] where each Bx independently represents any nucleobase.

[0400] In certain embodiments, an inverted nucleoside is terminal (i.e., the last nucleoside on one end of an oligonucleotide) and so only one internucleoside linkage depicted above will be present. In certain such embodiments, additional features (such as a conjugate group) can be attached to the inverted nucleoside. Such terminal inverted nucleosides can be attached to either or both ends of an oligonucleotide. In certain embodiments, such groups lack a nucleobase and are referred to herein as inverted sugar rnoieties. In certain embodiments, an inverted sugar moiety is terminal (i.e., attached to the last nucleoside on one end of an oligonucleotide) and so only one internucleoside linkage above will be present. In certain such embodiments, additional features (such as a conjugate group) can be attached to the inverted sugar moiety. PATENT 7328*158807PCT / NLM-1005-PC Such terminal inverted sugar rnoieties can be attached to either or both ends of an oligonucleotide.

[0401] In certain embodiments, nucleic acids can be linked 2’ to 5?rather than the standard 3’ to 5’ linkage. Such a linkage is illustrated below, where each Bx represents any nucleobase:

[0402]

[0403] Exemplary Motifs

[0404] In certain embodiments, modified oligonucleotides comprise one or more modified nucleosides comprising a modified sugar moiety. In certain embodiments, modified oligonucleotides comprise one or more modified nucleosides comprising a modified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more modified internucleoside linkage. In such embodiments, the modified, unmodified, and differently modified sugar rnoieties, nucleobases, and / or internucleoside linkages of a modified oligonucleotide define a pattern or motif. In certain embodiments, the patterns of sugar rnoieties, nucleobases, and internucleoside linkages are each independent of one another. Thus, a modified oligonucleotide can be described by its sugar motif, nucleobase motif and / or internucleoside linkage motif (as used herein, nucleobase motif describes the modifications to the nucleobases independent of the sequence of nucleobases).

[0405] Exemplary Sugar Motifs

[0406] In certain embodiments, oligonucleotides comprise one or more type of modified sugar and / or unmodified sugar moiety arranged along the oligonucleotide or region thereof in a defined pattern or sugar motif. In certain instances, such sugar motifs include but are not limited to any of the sugar modifications described herein. In certain embodiments, modified oligonucleotides comprise or consist of a region having a gapmer motif, which is defined by two external regions or "wings" and a central or internal region or “gap.” The three regions of a gapmer motif (the 5'-wmg PATENT 7328*158807PCT / NLM-1005-PC the central region (or gap), and the 3 ' -wing) form a contiguous sequence of nucleosides where at least some of the sugar moieties of the nucleosides of each of the wings differ from at least some of the sugar moieties of the nucleosides of the gap. Specifically, at least the sugar moieties of the nucleosides of each wing that are closest to the gap (the 3’-most nucleoside of the 5’-wing and the 5?-most nucleoside of the 3’-wing) differ from the sugar moiety of the neighboring gap nucleosides, thus defining the boundary between the wings and the gap (i.e., the wing / gap junction). In certain embodiments, the sugar moieties within the gap are the same as one another. In certain embodiments, the gap includes one or more nucleoside having a sugar moiety that differs from the sugar moiety of one or more other nucleosides of the gap. In certain embodiments, the sugar motifs of the two wings are the same as one another (symmetric gapmer). In certain embodiments, the sugar motif of the 5’ -Wing differs from the sugar motif of the 3’-wing (asymmetric gapmer).

[0407] In certain embodiments, the wings of a gapmer comprise 1-6 nucleosides. In certain embodiments, each nucleoside of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least one nucleoside of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least two nucleosides of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least three nucleosides of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least four nucleosides of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least five nucleosides of each wing of a gapmer comprises a modified sugar moiety.

[0408] In certain embodiments, the central region (or gap) of a gapmer comprises 7-12 nucleosides. In certain embodiments, each nucleoside of the gap of a gapmer comprises a 2’-beta-D-deoxyribosyl sugar moiety. In certain embodiments, at least three nucleosides, which sometimes are at least three contiguous nucleosides, of the central region of a gapmer comprise a 2’-beta-D-deoxyribosyl sugar moiety. In certain embodiments, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 nucleosides, or 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 contiguous nucleosides, in a central region each comprise a 2’-beta-D-deoxyribosyl sugar moiety. In certain embodiments, at least one nucleoside of the central region of a gapmer comprises a modified sugar moiety. In certain embodiments, one or two nucleosides of the central region of a gapmer comprises a modified sugar moiety. Certain embodiments in which a central region includes at PATENT 7328*158807PCT / NLM-1005-PC least one nucleoside comprising a modified sugar moiety and / or at least one modified internucleoside linkage are described herein.

[0409] In certain embodiments, the gapmer is a deoxy gapmer. In certain embodiments, the nucleosides on the gap side of each wing / gap junction compri se 2’-beta-D-deoxyribosyl sugar moieties and the nucleosides on the wing sides of each wing / gap junction comprise modified sugar moieties. In certain embodiments, each nucleoside of the gap comprises a 2 ’-beta-D-deoxy ribosyl sugar moiety. In certain embodiments, each nucleoside of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least one nucleoside of the gap of a gapmer comprises a modified sugar moiety. In certain embodiments, at least one nucleoside of the gap of a gapmer comprises a 2’-0Me sugar moiety or 2’ -MOE sugar moiety. In certain embodiments, modified oligonucleotides comprise or consist of a portion having a fully modified sugar motif. In such embodiments, each nucleoside of the fully modified portion of the modified oligonucleotide comprises a modified sugar moiety. In certain embodiments, each nucleoside of the entire modified oligonucleotide comprises a modified sugar moiety. In certain embodiments, modified oligonucleotides comprise or consist of a portion having a fully modified sugar motif, where each nucleoside within the fully modified portion comprises the same modified sugar moiety, referred to as a uniformly modified sugar motif. In certain embodiments, a fully modified oligonucleotide is a uniformly modified oligonucleotide. In certain embodiments, each nucleoside of a uniformly modified oligonucleotide comprises the same 2’-modification.

[0410] Herein, the lengths (number of nucleosides) of the three regions of a gapmer can be provided using the notation # of nucleosides in the 5’-wing] - # of nucleosides in the gap] - [# of nucleosides in the 3’-wingj. Thus, a 3-10-3 gapmer consists of 3 linked nucleosides in each wing and 10 linked nucleosides in the gap. Where such nomenclature is followed by a specific modification, that modification is the modification in each sugar moiety of each wing and the gap nucleosides comprise 2’-beta-D-deoxyribosyl sugar moieties. Thus, a 5-10-5 MOE gapmer consists of 5 linked 2 ’-MOE nucleosides in the 5 ’-wing, 10 linked 2’-beta-D-deoxynucleosides in the gap, and 5 linked 2’ -MOE nucleosides in the 3 ’-wing. A 3-10-3 cEt gapmer consists of 3 linked cEt nucleosides in the 5’-wing, 10 linked 2’-beta-D-deoxynucleosides in the gap, and 3 linked cEt nucleosides in the 3 ’-wing. A 5-8-5 PATENT 7328*158807PCT / NLM-1005-PC gapmer consists of 5 linked nucleosides comprising a modified sugar moiety in the 5’-wing, 8 linked 2’-beta-D-deoxynucleosides in the gap, and 5 linked nucleosides comprising a modified sugar moiety in the 3 ’-wing. A 5-8-5 mixed gapmer has at least two different modified sugar moieties in the 5’-wing and / or the 3’-wing.

[0411] In certain embodiments, modified oligonucleotides are 5-10-5 MOE gapmers. In certain embodiments, modified oligonucleotides are 3-10-3 BNA gapmers. In certain embodiments, modified oligonucleotides are 3-10-3 cEt gapmers. In certain embodiments, modified oligonucleotides are 3-10-3 LNA gapmers.

[0412] In certain embodiments, modified oligonucleotides have a sugar motif of 5’-eeeeeddddddddddeeeee-3’, where each "d " represents a 2’-beta-D-deoxyribosyl sugar moiety, each “e” represents a 2 ’-MOE sugar moiety.

[0413] Exemplary7Nucleobase Motifs

[0414] In certain embodiments, oligonucleotides comprise modified and / or unmodified nucleobases arranged along the oligonucleotide or region thereof in a defined pattern or motif. In certain embodiments, each nucleobase is modified. In certain embodiments, none of the nucleobases are modified. In certain embodiments, each purine or each pyrimidine is modified. In certain embodiments, each adenine is modified. In certain embodiments, each guanine is modified. In certain embodiments, each thymine is modified. In certain embodiments, each uracil is modified. In certain embodiments, each cytosine is modified. In certain embodiments, some or all of the cytosine nucleobases in a modified oligonucleotide are 5-methylcytosines. In certain embodiments, all of the cy tosine nucleobases are 5-methylcytosines and all of the other nucleobases of the modified oligonucleotide are unmodified nucleobases. In certain embodiments, modified oligonucleotides comprise a block of modified nucl eobases. In certain such embodiments, the block is at the 3 ’-end of the oligonucleotide. In certain embodiments the block is within 3 nucleosides of the 3’-end of the oli onucleotide. In certain embodiments, the block is at the 5 '-end of the oligonucleotide. In certain embodiments the block is within 3 nucleosides of the 5’-end of the oligonucleotide.

[0415] In certain embodiments, oligonucleotides having a gapmer motif comprise a nucleoside comprising a modified nucleobase. In certain such embodiments, one nucleoside comprising a modified nucleobase is in the central gap of an oligonucleotide having a gapmer motif. In certain such embodiments, the sugar PATENT 7328*158807PCT / NLM-1005-PC moiety of said nucleoside is a 2’-beta~D~deoxyribosyl sugar moiety. In certain embodiments, the modified nucleobase is selected from a 2-deoxypyrimidine and a 5-propynepyrimidine.

[0416]

[0417] In certain embodiments, oligonucleotides comprise modified and / or unmodified internucleoside linkages arranged along the oligonucleotide or region thereof in a defined pattern or motif. In certain embodiments, each internucleoside linking group is a phosphodiester internucleoside linkage (P=O). In certain embodiments, each internucleoside linking group of a modified oligonucleotide is a phosphorothioate internucleoside linkage (P=S). In certain embodiments, each internucleoside linkage of a modified oligonucleotide is independently selected from a phosphorothioate internucleoside linkage and phosphodiester internucleoside linkage. In certain embodiments, each phosphorothioate internucleoside linkage is independently selected from a stereo-random phosphorothioate a (Sp) phosphorothioate, and a (Rp) phosphorothioate.

[0418] In certain embodiments, the sugar motif of a modified oligonucleotide is a gapmer and the internucleoside linkages within the gap are all modified. In certain embodiments, some or all of the internucleoside linkages in the wings are unmodified phosphodiester internucleoside linkages. In certain embodiments, the terminal internucleoside linkages are modified. In certain embodiments, the sugar motif of a modified oligonucleotide is a gapmer, and the internucleoside linkage motif comprises at least one phosphodiester internucleoside linkage in at least one wing, where the at least one phosphodiester linkage is not a terminal internucleoside linkage, and the remaining internucleoside linkages are phosphorothioate internucleoside linkages. In certain such embodiments, all of the phosphorothioate linkages are stereo-random. In certain embodiments, all of the phosphorothioate linkages in the wings are (Sp) phosphorothi oates, and the gap comprises at least one Sp, Sp, or Rp motif. In certain embodiments, populations of modified oligonucleotides are enriched for modified oligonucleotides comprising such internucleoside linkage motifs.

[0419] In certain embodiments, modified oligonucleotides have an intemucieoside linkage motif of (from 5’ to 3’) sososssssssssssooss or sooosssssssssssooss, soooossssssssssooss or sssssssssssssssssss, where each “s"' represents a PATENT 7328*158807PCT / NLM-1005-PC phosphorothioate intemucleoside linkage and each “o’’ represents a phosphodiester internudeoside linkage. In certain embodiments, each “s” represents a (Sp)-phosphorothioate intemucleoside linkage.

[0420] In certain embodiments, modified oligonucleotides have an intemucleoside linkage motif comprising one or more mesyl phosphoramidate internudeoside linkages. In certain embodiments, one or more phosphorothioate internucleoside linkages or one or more phosphodiester internucleoside linkages of the internucleoside linkage motifs herein is substituted with a mesyl phosphoramidate internucleoside linkage.

[0421] Exemplary Lengths

[0422] It is possible to increase or decrease the length of an oligonucleotide without eliminating activity. For example, in Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of oligonucleotides 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. Oligonucleotides 25 nucleobases in length with 8 or 11 mismatch bases near the ends of the oligonucleotides were able to direct specific cleavage of the target RNA, albeit to a lesser extent than the oligonucleotides that contained no mismatches. Similarly, target specific cleavage was achieved using 13 nucleobase oligonucleotides, including those with 1 or 3 mismatches.

[0423] In certain embodiments, oligonucleotides (including modified oligonucleotides) can have any of a variety of ranges of lengths. In certain embodiments, oligonucleotides consist of X to Y linked nucleosides, where X represents the fewest number of nucleosides in the range and Y represents the largest number nucleosides in the range. In certain such embodiments, X and Y are each independently selected from 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50; provided that X< Y. For example, in certain embodiments, oligonucleotides consist of 12 to 13, 12 to 14, 12 to 15, 12 to 16, 12 to 17, 12 to 18, 12 to 19, 12 to 20, 12 to 21, 12 to 22, 12 to 23, 12 to 24, 12 to 25, 12 to 26, 12 to 27, 12 to 28, 12 to 29, 12 to 30, 13 to 14, 13 to 15, 13 to 16, 13 to 17, 13 to 18, 13 to 19, 13 to 20, 13 to 21, 13 to 22, 13 to 23, 13 to 24, 13 to 25, 13 to 26, 13 to 27, 13 to 28, 13 to 29, 13 to 30, 14 to 15, 14 to 16, 14 to 17, 14 to 18, 14 to 19, 14 to 20, 14 to 21, 14 to 22, 14 to 23, 14 to 24, 14 to 25, 14 to 26, 14 to 27, 14 to 28, 14 to 29, 14 to 30, PATENT 7328*158807PCT / NLM-1005-PC 15 to 16, 15 to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21, 15 to 22, 15 to 23, 15 to 24, 15 to 25, 15 to 26, 15 to 27, 15 to 28, 15 to 29, 15 to 30, 16 to 17, 16 to 18, 16 to 19, 16 to 20, 16 to 21, 16 to 22, 16 to 23, 16 to 24, 16 to 25, 16 to 26, 16 to 27, 16 to 28, 16 to 29, 16 to 30, 17 to 18, 17 to 19, 17 to 20, 17 to 21, 17 to 22, 17 to 23, 17 to 24, 17 to 25, 17 to 26, 17 to 27, 17 to 28, 17 to 29, 17 to 30, 18 to 19, 18 to 20, 18 to 21, 18 to 22, 18 to 23, 18 to 24, 18 to 25, 18 to 26, 18 to 27, 18 to 28, 18 to 29, 18 to 30, 19 to 20, 19 to 21, 19 to 22, 19 to 23, 19 to 24, 19 to 25, 19 to 26, 19 to 27, 19 to 28, 19 to 29, 19 to 30, 20 to 21, 20 to 22, 20 to 23, 20 to 24, 20 to 25, 20 to 26, 20 to 27, 20 to 28, 20 to 29, 20 to 30, 21 to 22, 21 to 23, 21 to 24, 21 to 25, 21 to 26, 21 to 27, 21 to 28, 21 to 29, 21 to 30, 22 to 23, 22 to 24, 22 to 25, 22 to 26, 22 to 27, 22 to 28, 22 to 29, 22 to 30, 23 to 24, 23 to 25, 23 to 26, 23 to 27, 23 to 28, 23 to 29, 23 to 30, 24 to 25, 24 to 26, 24 to 27, 24 to 28, 24 to 29, 24 to 30, 25 to 26, 25 to 27, 25 to 28, 25 to 29, 25 to 30, 26 to 27, 26 to 28, 26 to 29, 26 to 30, 27 to 28, 27 to 29, 27 to 30, 28 to 29, 28 to 30, or 29 to 30 linked nucleosides.

[0424] In certain embodiments, oligonucleotides (including modified oligonucleotides) consist of 16 linked nucleosides. In certain embodiments, oligonucleotides (including modified oligonucleotides) consist of 17 linked nucleosides. In certain embodiments, oligonucleotides (including modified oligonucleotides) consist of 18 linked nucleosides. In certain embodiments, oli onucleotides (including modified oligonucleotides) consist of 19 linked nucleosides. In certain embodiments, oligonucleotides (including modified oligonucleotides) consist of 20 linked nucleosides.

[0425] Exemplary Modified Oligonucleotides

[0426] In certain embodiments, the above modifications (sugar, nucleobase, internucleoside linkage) are incorporated into a modified oligonucleotide. In certain embodiments, modified oligonucleotides are characterized by their modification motifs and overall lengths. In certain embodiments, such parameters are each independent of one another. Thus, unless otherwise indicated, each internucleoside linkage of an oligonucleotide having a gapmer sugar motif can be modified or unmodified and may or may not follow the gapmer modification pattern of the sugar modifications. For example, the internucleoside linkages within the wing regions of a sugar gapmer can be the same or different from one another and can be the same or different from the internucleoside linkages of the gap region of the sugar motif. PATENT 7328*158807PCT / NLM-1005-PC Likewise, such sugar gapmer oligonucleotides may comprise one or more modified nucleobase independent of the gapmer pattern of the sugar modifications. Unless otherwise indicated, all modifications are independent of nucleobase sequence.

[0427] Certain oligonucleotide modifications can enhance antisense activity. An oligonucleotide may include one or more modifications described in Shen et al., Nature Biotechnology 640(37): 640–650 (2019); Migawa et al., Nucleic Acids Research 47(11): 5465–5479 (2019), Anderson et al., Nucleic Acids Research 49(16): 9026–9041 (2021) and Vasquez et al., Nucleic Acids Research 49(4): 1828–1839 (2021).

[0428] In certain embodiments, an oligomeric compound comprises a central region, and the central region comprises 1 to 5 (1, 2, 3, 4 or 5 for example) mesyl phosphoramidite and / or busyl phosphoramidite internucleoside linkages. In certain embodiments, the central region comprises a 5’ terminus, and position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises a mesyl phosphoramidite and / or a busyl phosphoramidite intemucleoside linkage. In certain embodiments, the central region comprises a 5’ terminus, and position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises a mesyl phosphoramidite internucleoside linkage.

[0429] In certain embodiments, an oligomeric compound comprises a central region, and the central region comprises 1 to 5 alkylphosphonate internucleoside linkages. In certain embodiments, the central region comprises a 5’ terminus; and position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises an alkylphosphonate internucleoside linkage. In certain embodiments, the central region comprises a 5’ terminus, and position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises a mesyl phosphoramidite and / or a busyl phosphoramidite intemucleoside linkage. In certain embodiments, the alkylphosphonate intemucleoside linkage comprises methoxypropyl (MOP). In certain embodiments, the alkylphosphonates intemucleoside linkage comprises methyl.

[0430] In certain embodiments, an oligomeric compound comprises a central region, and the central region comprises 1 to 5 (1, 2, 3, 4 or 5 for example) modified sugar PATENT 7328*158807PCT / NLM-1005-PC moieties. In certain embodiments, each of the 1 to 5 modified sugar moieties independently is a non-bicyclic modified sugar moiety. In certain embodiments, each of the 1 to 5 modified sugar moieties independently is a bicyclic modified sugar moiety. In certain embodiments, a modified sugar moiety comprises a 2’ substituent, and in certain instances is a 2’-0Me modified sugar moiety. In certain embodiments, a modified sugar moiety comprises a 5’ substituent, and in certain instances is a 5’-m ethyl or 5 ’-ethyl modified sugar moiety.

[0431] In certain embodiments, the central region comprises a 5’ terminus, and position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises a modified sugar moiety. In certain embodiments, position 2, or position 3, or positions 2 and 3 of the central region, as counted from the 5' terminus of the central region, each independently comprises a modified sugar moiety comprising a 2’ substituent. In certain embodiments, the modified sugar moiety independently is a 2’-0Me modified sugar moiety.

[0432] In certain embodiments, an oligomeric compound comprises a central region, the central region comprises a 5’ terminus; and position 3, or position 4, or positions 3 and 4 of the central region, as counted from the 5' terminus of the central region, each independently comprises a modified sugar moiety. In certain embodiments, position 3, or position 4, or positions 3 and 4 of the central region, as counted from the 5' terminus of the central region, each independently comprises modified sugar moiety comprising a 5’ substituent. In certain embodiments, position 3, or position 4, or positions 3 and 4 of the central region, as counted from the 5' terminus of the central region, each independently comprises a 5 ’-methyl modified sugar moiety. In certain embodiments, the 5 ’-methyl modified sugar moiety independently is a R-5’ -methyl modified sugar moiety or a S-5’ -methyl modified sugar moiety.

[0433]

[0434] of Modified Oligonucleotides

[0435] Populations of modified oligonucleotides in which all of the modified oligonucleotides of the population have the same molecular formula can be stereorandom populations or chirally enriched populations. All of the chiral centers of all of the modified oligonucleotides are stereo-random in a stereo-random population. In a chirally enriched population, at least one particular chiral center is not stereo-random in the modified oligonucleotides of the population. In certain embodiments, the PATENT 7328*158807PCT / NLM-1005-PC modified oligonucleotides of a chirally enriched population are enriched for beta-D ribosyl sugar moieties, and all of the phosphorothioate internucleoside linkages are stereo-random. In certain embodiments, the modified oligonucleotides of a chirally enriched population are enriched for both beta-D ribosyl sugar moieties and at least one, particular phosphorothioate internucleoside linkage in a particular stereochemical configuration.

[0436] Exemplary Nucleobase Sequences

[0437] In certain embodiments, oligonucleotides (unmodified or modified oligonucleotides) are further described by their nucleobase sequence. In certain embodiments oligonucleotides have a nucleobase sequence that is complementary to a second oligonucleotide or an identified reference nucleic acid, such as a target nucleic acid. In certain such embodiments, a region of an oligonucleotide has a nucleobase sequence that is complementary to a second oligonucleotide or an identified reference nucleic acid, such as a target nucleic acid. In certain embodiments, the nucleobase sequence of a region or entire length of an oligonucleotide is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% complementary to the second oligonucleotide or nucleic acid, such as a target nucleic acid.

[0438]

[0439] In certain embodiments, provided are oligomeric compounds, which consist of an oligonucleotide (modified or unmodified) and optionally one or more conjugate groups and / or terminal groups. Conjugate groups consist of one or more conjugate moiety and a conjugate linker which links the conjugate moiety to the oligonucleotide. Conjugate groups can be attached to either or both ends of an oligonucleotide and / or at any internal position. In certain embodiments, conjugate groups are attached to the 2'-position of a nucleoside of a modified oligonucleotide. In certain embodiments, conjugate groups that are attached to either or both ends of an oligonucleotide are terminal groups. In certain such embodiments, conjugate groups or terminal groups are attached at the 3’ and / or 5 ’-end of oligonucleotides. In certain such embodiments, conjugate groups (or terminal groups) are attached at the 3 "-end of oligonucleotides. In certain embodiments, conjugate groups are attached near the 3’- end of oligonucleotides. In certain embodiments, conjugate groups (or terminal groups) are attached at the 5" -end of oligonucleotides. In certain embodiments, PATENT 7328*158807PCT / NLM-1005-PC conjugate groups are attached near the 5 ’-end of oligonucleotides. Examples of terminal groups include but are not limited to conjugate groups, capping groups, phosphate moieties, protecting groups, modified or unmodified nucleosides, and two or more nucleosides that are independently modified or unmodified.

[0440] Exemplary Conjugate Groups

[0441] In certain embodiments, oligonucleotides are covalently attached to one or more conjugate groups. In certain embodiments, conjugate groups modify one or more properties of the attached oligonucleotide, including but not limited to pharmacodynamics, pharmacokinetics, stability, binding, absorption, tissue distribution, cellular distribution, cellular uptake, charge and clearance.

[0442] In certain embodiments, conjugation of one or more carbohydrate moieties to a modified oligonucleotide can optimize one or more properties of the modified oligonucleotide. In certain embodiments, the carbohydrate moiety is attached to a modified subunit of the modified oligonucleotide. For example, the ribose sugar of one or more ribonucleotide subunits of a modified oligonucleotide can be replaced with another moiety, for example, a non -carbohydrate (preferably cyclic) carrier to which is attached a carbohydrate ligand. A ribonucleotide subunit in which the ribose sugar of the subunit has been so replaced is referred to herein as a ribose replacement modification subunit (RRMS), which is a modified sugar moiety. A cyclic carrier can be a carbocyclic ring system, i.e., one or more ring atoms can be a heteroatom, for example, nitrogen, oxygen, sulfur. The cyclic carrier can be a monocyclic ring system, or may contain two or more rings, for example, fused rings. The cyclic carrier can be a fully saturated ring system, or it may contain one or more double bonds. In certain embodiments, the modified oligonucleotide is a gapmer.

[0443] In certain embodiments, conjugate groups impart a new property on the attached oligonucleotide, for example, fluorophores or reporter groups that enable detection of the oligonucleotide. Certain conjugate groups and conjugate moieties have been described previously, for example: cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053-1060), a thioether, for example, hexyl-S-tritylthiol (Manoharan et al., Ann. N. Y. Acad. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Lett, 1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain, for example, do-decan- PATENT 7328*158807PCT / NLM-1005-PC diol or undecyl residues (Saison-Behmoaras et al., EMBO J., 1991. 10, 1111-1118; Kabanov et al., FEES Lett., 1990, 259, 327-330; Svinarchuk et al, Biochimie, 1993, 75, 49-54), a phospholipid, for example, di-hexadecyl-rac-glycerol or triethyl-ammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et ah, Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res., 1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic acid a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Then, 1996, 277, 923-937), a tocopherol group (Nishina et al., Molecular Therapy Nucleic Acids, 2015, 4, e220, and Nishina et al., Molecular Therapy, 2008, 16, 734-740), or a GalNAc cluster (for example, WO2014 / 179620).

[0444] In certain embodiments, conjugate groups can be selected from any of a C22 alkyl, C20 alkyl, C16 alkyl, CIO alkyl, C21 alkyl, C19 alkyl, C18 alkyl, C17 alkyl, C15 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, CH alkyl, C9 alkyl, C8 alkyl. Cl alkyl, C6 alkyl, C5 alkyl, C22 alkenyl, C20 alkenyl, C16 alkenyl, CIO alkenyl, C21 alkenyl, C19 alkenyl, Cl 8 alkenyl, C17 alkenyl, Cl 5 alkenyl, C14 alkenyl, C13 alkenyl, C12 alkenyl, Cl 1 alkenyl, C9 alkenyl, C8 alkenyl, Cl alkenyl, C6 alkenyl, or C5 alkenyl. In certain embodiments, conjugate groups can be selected from any of C22 alkyl, C20 alkyl, Cl 6 alkyl, CIO alkyl, C21 alkyl, Cl 9 alkyl, C18 alkyl, C17 alkyl. Cl 5 alkyl, C14 alkyl. Exemplary C13 alkyl, C12 alkyl, CH alkyl, C9 alkyl, C8 alkyl, C7 alkyl, C6 alkyl, and C5 alkyl, where the alkyl chain has one or more unsaturated bonds. In certain embodiments, a conjugate group is a lipid having the following structure:

[0445]

[0446] In certain embodiments, a conjugate moiety comprises an active drug substance, for example, aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen, dansyl sarcosine, 2,3,5-triiodobenzoic acid, fingolimod, flufenamic acid, folinic acid, a benzothiadi azide, chlorothiazide, a diazepine, indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, an antidiabetic, an antibacterial or an antibiotic.

[0447] Conjugate moieties can include, without, limitation, intercalators, reporter molecules, poly mines, polyamides, peptides, carbohydrates (e g., GalNAc), PATENT 7328*158807PCT / NLM-1005-PC antibodies, vitamin moieties, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipids, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins, fluorophores, and dyes.

[0448] Exemplary Conjugate Linkers

[0449] Conjugate moieties are attached to oligonucleotides through conjugate linkers. In certain oligomeric compounds, the conjugate linker is a single chemical bond (i.e., the conjugate moiety is attached directly to an oligonucleotide through a single bond). In certain embodiments, the conjugate linker comprises a chain structure, such as a hydrocarbyl chain, or an oligomer of repeating units such as ethylene glycol, nucleosides, or amino acid units.

[0450] In certain embodiments, a conjugate linker comprises pyrrolidine. In certain embodiments, a conjugate linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In certain such embodiments, the conjugate linker comprises groups selected from alkyl, amino, oxo, amide and ether groups. In certain embodiments, the conjugate linker comprises groups selected from alkyl and amide groups. In certain embodiments, the conjugate linker comprises groups selected from alkyl and ether groups. In certain embodiments, the conjugate linker comprises at least one phosphorus moiety. In certain embodiments, the conjugate linker comprises at least one phosphate group or phosphodi ester group. In certain embodiments, the conjugate linker includes at least one neutral linking group.

[0451] In certain embodiments, conjugate linkers, including the conjugate linkers described above, are bifunctional linking moieties, for example, those known in the art to be useful for attaching conjugate groups to compounds, such as the oligonucleotides provided. In general, a bifunctional linking moiety comprises at least two functional groups. One of the functional groups is selected to bind to a particular site on a compound and the other is selected to bind to a conjugate group. Examples of functional groups used in a bifunctional linking moiety include but are not limited to electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In certain embodiments, bifunctional linking moieties comprise one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl. PATENT 7328*158807PCT / NLM-1005-PC Examples of conjugate linkers include but are not limited to pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane"! “Carboxyl ate (SMCC) and 6-aminohexanoic acid (AHX or AHA). Other conjugate linkers include but are not limited to substituted or unsubstituted Cl-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl or substituted or unsubstituted C2-C10 alkynyl, where a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.

[0452] In certain embodiments, conjugate linkers comprise 1-10 linker-nucleosides. In certain embodiments, conjugate linkers comprise 2-5 linker-nucleosides. In certain embodiments, conjugate linkers comprise exactly 3 linker-nucleosides. In certain embodiments, conjugate linkers comprise the TCA motif. In certain embodiments, such linker-nucleosides are modified nucleosides. In certain embodiments such linker-nucleosides comprise a modified sugar moiety. In certain embodiments, linker-nucleosides are unmodified. In certain embodiments, linker-nucleosides comprise an optionally protected heterocyclic base selected from a purine, substituted purine, pyrimidine or substituted pyrimidine. In certain embodiments, a cleavable moiety is a nucleoside selected from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methylcytosine, 4-N-benzoyl-5-methylcytosine, adenine, 6-N-benzoyladenine, guanine and 2-N-isobutyrylguanine. It is typically desirable for linker-nucleosides to be cleaved from the oligomeric compound after it reaches a target tissue. Accordingly, linker-nucleosides are typically linked to one another and to the remainder of the oligomeric compound through cleavable bonds. In certain embodiments, such cleavable bonds are phosphodiester bonds.

[0453] Herein, linker-nucleosides are not considered to be part of the oligonucleotide. Accordingly, in embodiments in which an oligomeric compound comprises an oligonucleotide consisting of a specified number or range of linked nucleosides and / or a specified percent complementarity to a reference nucleic acid and the oligomeric compound also comprises a conjugate group comprising a conjugate linker comprising linker-nucleosides, those linker-nucleosides are not counted toward the length of the oligonucleotide and are not used in determining the percent complementarity of the oligonucleotide for the reference nucleic acid. For example, an oligomeric compound may comprise (1) a modified oligonucleotide consisting of PATENT 7328*158807PCT / NLM-1005-PC 8-30 nucleosides and (2) a conjugate group comprising 1-10 linker-nucleosides that are contiguous with the nucleosides of the modified oligonucleotide. The total number of contiguous linked nucleosides in such an oligomeric compound is more than 30. Alternatively, an oligomeric compound may comprise a modified oligonucleotide consisting of 8-30 nucleosides and no conjugate group. The total number of contiguous linked nucleosides in such an oligomeric compound is no more than 30. Unless otherwise indicated conjugate linkers comprise no more than 10 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 5 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 3 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 2 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than I linker-nucleoside.

[0454] In certain embodiments, it is desirable for a conjugate group to be cleaved from the oligonucleotide. For example, in certain circumstances oligomeric compounds comprising a particular conjugate moiety are better taken up by a particular cell type, but once the oligomeric compound has been taken up, it is desirable that the conjugate group be cleaved to release the unconjugated or parent oligonucleotide. Thus, certain conjugate linkers may comprise one or more cleavable moieties. In certain embodiments, a cleavable moiety is a cleavable bond. In certain embodiments, a cleavable moiety is a group of atoms comprising at least one cleavable bond. In certain embodiments, a cleavable moiety comprises a group of atoms having one, two, three, four, or more than four cleavable bonds. In certain embodiments, a cleavable moiety is selectively cleaved inside a cell or subcellular compartment, such as a lysosome. In certain embodiments, a cleavable moiety is selectively cleaved by endogenous enzymes, such as nucleases.

[0455] In certain embodiments, a cleavable bond is selected from among an amide, an ester, an ether, one or both esters of a phosphodi ester, a phosphate ester, a carbamate, or a disulfide. In certain embodiments, a cleavable bond is one or both of the esters of a phosphodiester. In certain embodiments, a cleavable moiety comprises a phosphate or phosphodiester. In certain embodiments, the cleavable moiety is a phosphate linkage or phosphodiester linkage between an oligonucleotide and a conjugate moiety or conjugate group. PATENT 7328*158807PCT / NLM-1005-PC In certain embodiments, a cleavable moiety comprises or consists of one or more linker-nucleosides. In certain such embodiments, the one or more linker-nucleosides are linked to one another and / or to the remainder of the oligomeric compound through cleavable bonds. In certain embodiments, such cleavable bonds are unmodified phosphodiester bonds. In certain embodiments, a cleavable moiety is 2'-deoxynudeoside that is attached to either the 3’ or 5'-terminal nucleoside of an oligonucleotide by a phosphate internucleoside linkage or phosphodiester internucleoside linkage and covalently attached to the remainder of the conjugate linker or conjugate moiety by a phosphate, phosphodiester or phosphorothioate linkage. In certain such embodiments, the cleavable moiety is 2' -deoxy adenosine. Cell- Targeting Moieties

[0456] In certain embodiments, a conjugate group comprises a cell-targeting moiety. In certain embodiments, a conjugate group has the general formula:

[0457] Cleavable Linker Moiety

[0458]

[0459] Cell-targeting moiety Conjugate Linker

[0460] where n is from 1 to about 3, m is 0 when n is 1, m is 1 when n is 2 or greater, j is 1 or 0, and k is 1 or 0. In certain embodiments, n is 1, j is 1 and k is 0. In certain embodiments, n is I, j is 0 and k is I. In certain embodiments, n is 1, j is I and k is 1. In certain embodiments, n is 2, j is 1 and k is 0. In certain embodiments, n is 2, j is 0 and k is 1. In certain embodiments, n is 2, j is 1 and k is I. In certain embodiments, n is 3, j is I and k is 0. In certain embodiments, n is 3, j is 0 and k is 1. In certain embodiments, n is 3, j is 1 and k is 1.

[0461] In certain embodiments, conjugate groups comprise cell -targeting moieties that have at least one tethered ligand. In certain embodiments, cell -targeting moieties comprise two tethered ligands covalently attached to a branching group. In certain embodiments, cell-targeting moieties comprise three tethered ligands covalently attached to a branching group.

[0462] In certain embodiments, each ligand of a cell-targeting moiety has an affinity for at least one type of receptor on a target cell. In certain embodiments, each ligand has an affinity for at least one type of receptor on the surface of a mammalian liver cell. In certain embodiments, each ligand has an affinity for the hepatic asialoglycoprotein receptor (ASGP-R). In certain embodiments, each ligand is a PATENT 7328*158807PCT / NLM-1005-PC carbohydrate. In certain embodiments, a conjugate group comprises a cell-targeting conjugate moiety. In certain embodiments, a conjugate group has the general formula:

[0463] [Conjugate Linker }r~ i Cleavable Conj.

[0464] Moiety I Linker Moiety

[0465]

[0466]

[0467]

[0468]

[0469] Cell-targeting conjugate moiety Conjugate Linker

[0470] where n is from 1 to about 3, m is 0 when n is 1, m is 1 when n is 2 or greater, j is 1 or 0, and k is 1 or 0. In certain embodiments, n is 1, j is I and k is 0. In certain embodiments, n is 1, j is 0 and k is 1. In certain embodiments, n is 1, j is 1 and k is 1. In certain embodiments, n is 2, j is 1 and k is 0. In certain embodiments, n is 2, j is 0 and k is 1. In certain embodiments, n is 2, j is I and k is 1. In certain embodiments, n is 3, j is 1 and k is 0. In certain embodiments, n is 3, j is 0 and k is I. In certain embodiments, n is 3, j is 1 and k is 1.

[0471] In certain embodiments, conjugate groups comprise cell -targeting moi eties that have at least one tethered ligand. In certain embodiments, cell-targeting moieties comprise two tethered ligands covalently attached to a branching group. In certain embodiments, cell-targeting moieties comprise three tethered ligands covalently attached to a branching group.

[0472] In certain embodiments, the cell-targeting moiety targets neurons. In certain embodiments, the cell-targeting moiety targets a neurotransmitter receptor. In certain embodiments, the ceil targeting moiety targets a neurotransmitter transporter. In certain embodiments, the cell targeting moiety targets a GABA transporter. See for example, WO 2011 / 131693, WO 2014 / 064257.

[0473] In certain embodiments, conjugate groups comprise cell-targeting moieties that have affinities for transferrin receptor (TfR.) (also referred to herein as TfRl and CD71). In certain embodiments, a conjugate group described herein comprises an anti-TfRl antibody or fragment thereof. In certain embodiments, the conjugate group comprises a protein or peptide capable of binding TfRl. In certain embodiments, the conjugate group comprises an aptamer capable of binding TfRl. In certain embodiments, the anti-TfRl antibody or fragment thereof can be any known in the art including but not limited to those described in WO1991 / 004753; W02013 / 103800; W02014 / 144060; WO2016 / 081643; WO2016 / 179257; WO2016 / 207240;

[0474] WO2017 / 221883; WO2018 / 129384, WO2018 / 124121; WO2019 / 151539;

[0475] WO2020 / 132584; W02020 / 028864; US 7,208,174; US 9,034,329; and US PATENT 7328*158807PCT / NLM-1005-PC 10,550,188. In certain embodiments, a fragment of an anti-TiRl antibody is F(ab‘)2, Fab, Fab', Fv, or scFv.

[0476] In certain embodiments, the conjugate group comprises a protein or peptide capable of binding TfRI. In certain embodiments, the protein or peptide capable of binding TfRI can be any known in the art including but not limited to those described in W02019 / 140050; W02020 / 037150; W02020 / 124032; and US 10,138,483.

[0477] In certain embodiments, the conjugate group comprises an aptamer capable of binding TfRI. In certain embodiments, the aptamer capable of binding TfRI can be any known in the art including but not limited to those described in WO2013 / 163303; WO2019 / 033051; and WO2020 / 245198.

[0478] In certain embodiments, a conjugate group includes a targeting moiety and a linker moiety. In certain embodiments, a targeting moiety comprises a carbohydrate ligand. In certain embodiments, a carbohydrate ligand in a conjugate group has affinity for an asialoglycoprotein receptor (ASGPR). In certain embodiments, a carbohydrate ligand in a conjugate group can target an oligomeric compound conjugate to liver cells and / or hepatocytes. In certain embodiments, a carbohydrate ligand includes a N-acetylgalactosamine (GalNAc) moiety. Non-limiting examples of GalNAc-containing targeting moieties and associated linker moieties that can be included in applicable conjugate groups are described in WO2014 / 179629.

[0479] In certain embodiments, a targeting moiety of a conjugate group includes two, three or four GalNAc ligand moieties. In certain embodiments, a targeting moiety containing multiple GalNAc moieties includes a branching group (also referred to as a branching moiety) and an individual tether moiety linking the branching group to each GalNAc ligand moiety. In certain embodiments a targeting moiety is cleavable. A branching moiety is cleavable in certain embodiments. Non-limiting examples of GalNAc ligand moieties, branching groups and tether moieties are described in WO2014 / 179629.

[0480] In certain embodiments, a cleavable targeting moiety comprises or consists of a structure according to the following general Formula A: PATENT 7328*158807PCT / NLM-1005-PC

[0481]

[0482] Formula A where each m and each n independently is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12. In certain embodiments pertaining to a targeting moiety according to general Formula A, each m independently is an integer of 5, 6 or 7 and each n independently is an integer of 1, 2 or 3. In certain embodiments pertaining to a targeting moiety of general Formula A, each m is the integer 6 and each n is the integer 2.

[0483] In certain embodiments, a linker of an oligonucleotide conjugate containing a targeting moiety joins the targeting moiety to the oligonucleotide. In certain embodiments, a linker moiety comprises or consists of a structure according to one of the following general Formula B, Formula C or Formula D:

[0484]

[0485] PATENT 7328*158807PCT / NLM-1005-PC where each n in Formula B or Formula C independently is zero or an integer of 1, 2, 3, 4, 5, 6 or 7. In certain embodiments, each n in Formula B or Formula C independently is an integer of 1, 2, 3, 4 or 5, and in certain embodiments each n in Formula B or Formula D independently is the integer 2 or 5. In certain embodiments, the terminal linker portion on the right-hand side of the linker moiety according to general Formula B, Formula C or Formula D can be, or can be a portion of, a phosphodiester group of a conjugate. In certain embodiments, a phosphodiester group is linked to the 5’ end of the 5’ terminal nucleoside of an oligonucleotide, sometimes via a methylene group linked to a sugar moiety of the 5’ terminal nucleoside of the oligonucleotide, for example. A non-limiting example of such a phosphodiester linkage between a linker moiety and oligonucleotide of a conjugate is depicted in the GalNAc-ASOl study drug structure according to the general formula in Example 2. In certain embodiments, a conjugate including a targeting moiety and linker moiety

[0486]

[0487] where the terminal linker portion on the right-hand side of the molecule as depicted can be, or can be a portion of, a phosphodiester moiety of a oligomeric compound conjugate. In certain embodiments, a phosphodiester moiety is linked to the 5’ end of the 5’ terminal nucleoside of an oligonucleotide, sometimes via a methylene group linked to a sugar moiety of the 5’ terminal nucleoside of the oligonucleotide, for example (see the GalNAc-ASOl study drug structure according to the general formula in Example 2 as a non-limiting example).

[0488] Certain Terminal Groups

[0489] In certain embodiments, oligomeric compounds comprise one or more terminal groups. In certain such embodiments, oligomeric compounds comprise a stabilized 5 ’-phosphate. Stabilized 5’~phosphates include, but are not limited to 5’- PATENT 7328*158807PCT / NLM-1005-PC phosphonates, including, but not limited to 5’-vinylphosphonates. In certain embodiments, terminal groups comprise one or more abasic sugar moi eties and / or inverted nucleosides. In certain embodiments, terminal groups comprise one or more 2’-linked nucleosides or sugar moieties. In certain such embodiments, the 2’-linked group is an abasic sugar moiety.

[0490] Antisense Activity

[0491] In certain embodiments, oligomeric compounds and oligomeric duplexes are capable of hybridizing to a target nucleic acid, resulting in at least one antisense activity and such oligomeric compounds ami oligomeric duplexes are referred to as "‘antisense compounds.’" In certain embodiments, antisense compounds have antisense activity when they reduce or inhibit the amount or activity of a target nucleic acid by 25% or more in the standard in vitro assay. In certain embodiments, antisense compounds selectively affect one or more target nucleic acid. Such antisense compounds comprise a nucleobase sequence that hybridizes to one or more target nucleic acid, resulting in one or more desired antisense activity, and does not hybridize to one or more non-target nucleic acid or does not hybridize to one or more non-target nucleic acid in such a way that results in significant undesired antisense activity.

[0492] For certain antisense activities, hybridization of an antisense compound to a target nucleic acid results in recruitment of a protein that cleaves the target nucleic acid. For example, certain antisense compounds result in RNase H mediated cleavage of the target nucleic acid. RNase H is a cellular endonuclease that cleaves the RNA strand of an RNA. DN duplex. The DNA in such an RNA: DNA duplex need not be unmodified DNA and can include modified oligonucleotide of antisense compounds. In certain embodiments, antisense compounds are sufficiently "‘DNA-iike” to elicit RNase H activity. In certain embodiments, one or more non-DNA-like nucleoside in the gap of a gapmer is tolerated.

[0493] For certain antisense activities, an antisense compound or a portion of an antisense compound is loaded into an RNA-induced silencing complex (RISC), ultimately resulting in cleavage of the target nucleic acid. For example, certain antisense compounds result in cleavage of the target nucleic acid by Argonaute. Antisense compounds that are loaded into RISC are RNAi agents. RNAi agents can be double-stranded (siRNA or dsRNAi) or single-stranded (ssRNAi). PATENT 7328*158807PCT / NLM-1005-PC In certain embodiments, hybridization of an antisense compound to a target nucleic acid does not result in recruitment of a protein that cleaves that target nucleic acid. In certain embodiments, hybridization of the antisense compound to the target nucleic acid results in alteration of splicing of the target nucleic acid. In certain embodiments, hybridization of an antisense compound to a target nucleic acid results in inhibition of a binding interaction between the target nucleic acid and a protein or other nucleic acid. In certain embodiments, hybridization of an antisense compound to a target nucleic acid results in alteration of translation of the target nucleic acid.

[0494] An antisense activity can be observed directly or indirectly. In certain embodiments, observation or detection of an antisense activity involves observation or detection of a change in an amount of a target nucleic acid or protein encoded by such target nucleic acid, a change in the ratio of splice variants of a nucleic acid or protein and / or a phenotypic change in a cell or subject.

[0495] Exemplary Target Nucleic Acids

[0496] In certain embodiments, oligomeric compounds comprise or consist of an oligonucleotide comprising a region that is complementary to a target nucleic acid. In certain embodiments, the target nucleic acid is an endogenous RNA molecule. In certain embodiments, the target nucleic acid encodes a protein. In certain such embodiments, the target nucleic acid is selected from: a mature mRNA and a pre-mRNA, including intronic, exonic and untranslated regions. In certain embodiments, the target RNA is a mature mRNA. In certain embodiments, the target nucleic acid is a pre-mRNA. In certain embodiments, the target region is entirely within an intron. In certain embodiments, the target region spans an intron / exon junction. In certain embodiments, the target region is at least 50% within an intron. In certain embodiments, the target nucleic acid is the RNA transcriptional product of a retrogene. In certain embodiments, the target nucleic acid is a non-coding RNA. In certain embodiments, the target non-coding RNA is selected from: a long non-coding RNA, a short non-coding RNA, an intronic RNA molecule.

[0497] Complementarity / Mismatches to Target Nucleic Acid and Duplex Complementarity In certain embodiments, oligonucleotides are complementary to the target nucleic acid over the entire length of the oligonucleotide. In certain embodiments, oligonucleotides are 99%, 95%, 90%, 85%, or 80% complementary to the target nucleic acid. In certain embodiments, oligonucleotides are at least 80% PATENT 7328*158807PCT / NLM-1005-PC complementary to the target nucleic acid over the entire length of the oligonucleotide and comprise a region that is 100% or fully complementary to a target nucleic acid. In certain embodiments, the region of full complementarity is from 6 to 20, 10 to 18, or 18 to 20 nucleobases in length.

[0498] It is possible to introduce mi match bases without eliminating activity. For example, Gautschi et al. (J. Natl. Cancer Inst. 93:463-471, March 2001) demonstrated the ability of an oligonucleotide having 100% complementarity to the bcl-2 mRNA and having 3 mismatches to the bcl-xL mRNA to reduce the expression of both bcl-2 and bcl-xL in vitro and in vivo. Furthermore, this oligonucleotide demonstrated potent anti-tumor activity in vivo. Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988) tested a series of tandem 14 nucleobase oligonucleotides, and 28 and 42 nucleobase oligonucleotides comprised of the sequence of two or three of the tandem oligonucleotides, respectively, for their ability to arrest translation of human DHFR in a rabbit reticulocyte assay. Each of the three 14 nucleobase oligonucleotides alone was able to inhibit translation, albeit at a more modest level than the 28 or 42 nucleobase oligonucleotides.

[0499] In certain embodiments, oligonucleotides comprise one or more mismatched nucleobases relative to the target nucleic acid. In certain embodiments, anti sense activity against the target is reduced by such mismatch, but activity against a nontarget is reduced by a greater amount. Thus, in certain embodiments selectivity of the oligonucleotide is improved.

[0500] In certain embodiments, a mismatch is specifically positioned within an oligonucleotide having a gapmer motif. In certain embodiments, the mismatch is at position 1, 2, 3, 4, 5, 6, 7, or 8 from the 5 ’-end of the gap region (central region). In certain embodiments, the mismatch is at. position 9, 8, 7, 6, 5, 4, 3, 2, 1 from the d’end of the gap region. In certain embodiments, the mismatch is at position 1, 2, 3, or 4 from the 5 ’-end of the wing region. In certain embodiments, the mismatch is at position 4, 3, 2, or 1 from the 3 ’-end of the wing region.

[0501] SAA1

[0502] In certain embodiments, oligomeric compounds reduce serum amyloid Al (SAA1) expression. In certain embodiments, an oligomeric compound comprises or consists of an oligonucleotide comprising a region that is complementary to a target region in a SAA1 nucleic acid, and thereby targets the SAA1 nucleic acid. PATENT 7328*158807PCT / NLM-1005-PC In certain embodiments, contacting a cell with an oligomeric compound targeted to a SAA1 nucleic acid target region reduces the amount of SAA1 RNA, and in certain embodiments reduces the amount of SAA1 protein. In certain embodiments, the oligomeric compound consists of a modified oligonucleotide. In certain embodiments, the oligomeric compound consists of a modified oligonucleotide and a conjugate group.

[0503] In certain embodiments, contacting a cell with an oligomeric compound targeted to a SAA1 nucleic acid target region reduces the amount of SAA1 RNA in a cell. In certain embodiments, contacting a cell with an oligomeric compound targeted to a SAA1 nucleic acid target region reduces the amount of SAA1 protein in the cell. In certain embodiments, the cell is in vitro. In certain embodiments, the cell is in a subject. In certain embodiments, the oligomeric compound consists of a modified oligonucleotide. In certain embodiments, contacting a cell in a subject with an oligomeric compound targeted to a SAA1 nucleic acid target region ameliorates one or more symptoms or hallmarks of a SAA1 -associated medical condition. In certain embodiments, the medical condition is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia. In certain embodiments, the symptom or hallmark is proteinuria, nephrotic syndrome, low blood protein level, low blood albumin level, edema, high cholesterol, enlarged spleen, enlarged liver, enlarged thyroid, orthostatic hypotension, gastrointestinal atony, diarrhea and constipation.

[0504] In certain embodiments, an oligomeric compound targeted to a SAA1 nucleic acid target region is capable of reducing the amount of SAA1 RN A in vitro by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% when administered according to the standard in vitro assay. In certain embodiments, an oligomeric compound targeted to a SAA1 nucleic acid target region is capable of reducing the amount of SAA1 RNA in vivo by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% when administered according to a standard in vivo assay. In certain embodiments, an oligomeric compound targeted to a SAA1 nucleic acid target region is capable of reducing the amount of SAA1 protein in vitro by at least 10%, at. least. 20%, at least 30%, at least 40%, at. least. 50%, at least 60%, at least 70%, at least 80%, or at least 90% when administered according to a standard in vitro assay. PATENT 7328*158807PCT / NLM-1005-PC In certain embodiments, an oligomeric compound targeted to a SAA1 nucleic acid target region is capable of reducing the amount of SAA1 protein in vivo by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% when administered according to a standard in vivo assay. In certain embodiments, an oligomeric compound targeted to a SAA1 nucleic acid target region is capable of reducing the amount of SAA1 RNA in a cell of a subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In certain embodiments, an oligomeric compound targeted to a SAA1 nucleic acid target region is capable of reducing the amount of SAA1 protein or the amount of SAA1 protein in a cell of a subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. Suitable cells include liver cells, and hepatocytes in certain embodiments, for example. In certain embodiments, SAA1 expression in a cell, tissue or subject is determined in vitro, ex vivo or in vivo. A standard in vitro assay is the quantitative polymerase chain reaction (PCR) assay described in Example 1, with a suitable type of cell.

[0505] SAA1 is located on chromosome 11 and Table A provides access information for a human SAA1 genomic reference sequence (Chromosome 11: 18,266,260-18,269,977; database reference accession no. ENSG00000173432; Genome Assembly GRCh38, release 112; SEQ ID NO:1), SAA1 complementary DNA(cDNA) sequence (database reference accession no. ENST00000356524.9; Genome Assembly GRCh38, release 112; SEQ ID NO:2) and SAA1 protein sequence (database reference accession no. ENSP00000348918.4; Genome Assembly GRCh38, release 112; SEQ ID NO: 14). A SAA1 coding sequence (CDS) within the cDNA sequence of SEQ ID NO:2 is provided as SEQ ID NO:3. Each of the genomic, cDNA and protein sequence referenced in Table A is from Genome Assembly GRCh38, release 112 (May 2024), accessible at World Wide Web URL useast.ensembl.org / . Table A also provides exon and intron and exon boundaries within the SAA1 reference genomic sequence (SEQ ID NO:1).

[0506] Table A: Homo sapiens SAA1 sequences and subregions

[0507]

[0508] PATENT 7328*158807PCT / NLM-1005-PC

[0509]

[0510] In certain embodiments, a target region in a SAA1 nucleic acid to which an oligomeric compound is targeted is 100% identical to an equal length portion in the SAA1 reference genomic sequence set forth in SEQ ID NO: 1 or SAA1 cDNA sequence set forth in SEQ ID NO: 2, In certain embodiments, a SAA1 target region to which an oligomeric compound is targeted is at least 80% identical to, or at least 85% identical to, or at least 90% identical to, or at least 95% identical to, an equal length portion in the SAA1 reference genomic sequence set forth in SEQ ID NO: I or SAA1 cDNA sequence set forth in SEQ ID NO: 2. In certain embodiments, a target region to which an oligomeric compound is targeted is identical to an equal length portion in the SAA1 reference genomic sequence set forth in SEQ ID NO: 1 or SAA1 cDNA sequence set forth in SEQ ID NO: 2 except for a one-nucleobase mismatch. In certain embodiments, an oligomeric compound oligonucleotide comprises or consists of a nucleobase sequence that is at least 80% complementary to, or at least 85% complementary' to, or at least 90% complementary to, or at least 95% complementary to, an equal length portion in the SAA1 reference genomic sequence set forth in SEQ ID NO: 1 or SAA1 cDNA sequence set forth in SEQ ID NO: 2.

[0511] In certain embodiments, an oligomeric compound oligonucleotide comprises or consists of nucleobase sequence containing at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases complementary to: (i) an equal length portion of nucleobases 3458-3708, 3624-3727 or 3644-3707 of SEQ ID NO:1; or (ii) an equal length portion of nucleobases 3644-3663, 3647-3666, 3652-3671, 3663-3682, 3666-3685, 3677-3696 or 3688-3707 of SEQ ID NO:1; or (iii) an equal length portion of nucleobases 720-3074, 2908-2967 or 2928-2947 of SEQ ID NO:1; or (iv) PATENT 7328*158807PCT / NLM-1005-PC an equal length portion of nucleobases 720-2935, 2200-2259 or 2220-2239 of SEQ ID NO:1; or (v) an equal length portion of nucleobases 3643-3702, 3643-3692 or 3663-3682 of SEQ ID NO: 1.

[0512] In certain embodiments, a SAA1 nucleic acid comprises a pathogenic genetic variant associated with a medical condition at a pathogenic genetic variation position and / or a pathogenic genetic variation site. In certain embodiments, the medical condition is a SAA1 -associated medical condition, and in certain instances is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia. In certain instances, a pathogenic genetic variation position and / or a pathogenic genetic variation site comprises a single nucleotide variation (SNV), and sometimes is a non-synonymous SNV located in an exon of a mutant SAA1 nucleic acid. In certain embodiments, a pathogenic genetic variant results in an amino acid modification (an amino acid substitution for example) in a SAA1 protein selected from p. Leu3Pro, p. Val8Ala, p. Asp34Val, p. Ala70Val, p. Trp71Arg, p. Val75Ala, р. Asp93Glu, p. Gly90Asp, p. Asn101Asp and p. Lys121Arg. In certain embodiments, a pathogenic genetic variant is chosen from c.8T> C, c.23T> C, c.209C> T, c.101 A> T, с.211T> C, c.224T> C, c.279C>T, c.269G> A, c.288T> C, c.301A> G orc.362A> C (positions are according to the SAA1 coding sequence (CDS; SEQ ID NO:3). In certain embodiments, a pathogenic genetic variation position and / or a pathogenic genetic variation site occurs in an exon, and optionally exon 3. In certain embodiments, a SAA1 nucleic acid comprises s pathogenic genetic variant that results in a Asp34Val amino acid substitution in a SAA1 protein (SEQ ID NO: 14 for example), and optionally is c.101 A> T. The Asp34Val amino acid substitution may result in misfolding, precipitation, and aggregation of SAA1.

[0513] In certain embodiments, an oligomeric compound oligonucleotide comprises or consists of a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of a nucleobase sequence selected from: SEQ ID NO:4 to SEQ ID NO: 12. In certain embodiments, an oligomeric compound oligonucleotide comprises or consists of a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of a nucleobase sequence selected from: SEQ ID NO:4 to SEQ ID NO:7 and SEQ ID NO:9 to SEQ ID PATENT 7328*158807PCT / NLM-1005-PC NO: 11. In certain embodiments, an oligomeric compound oligonucleotide comprises or consists of a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of SEQ ID NO: 12. In certain embodiments, an oligomeric compound oligonucleotide comprises or consists of a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of a nucleobase sequence selected from: SEQ ID NO: 8.

[0514] In certain embodiments, a modified oligonucleotide of a oligomeric compound consists of 18-22 linked nucleosides, 19-21 linked nucleosides or 20 linked nucleosides. In certain embodiments, a modified oligonucleotide of a oligomeric compound includes at least one modification selected from a modified sugar moiety, or a modified internucleoside linkage, or a modified sugar moiety and a modified internucleoside linkage.

[0515] In certain embodiments, a modified oligonucleotide is a gapmer that includes a 5'-region, a central region and a 3’-region. In certain embodiments, a modified oligonucleotide of an oligomeric compound is a MOE gapmer. In certain embodiments, a modified oligonucleotide is a 5-10-5 MOE gapmer. In certain embodiments, the sugar motif for the gapmers is (from 'n to 3’) eeeeeddddddddddeeeee, where each “d” represents a 2' -beta-D-deoxy ribosyl sugar moiety, and each “e” represents a 2’- MOE sugar moiety. In certain embodiments, the internucleoside linkages of a modified oligonucleotide are phosphorothioate internucleoside linkages and phosphodi ester internucleoside linkages. In certain embodiments, a modified oligonucleotide comprises or consists of internucleoside linkages according to the linkage motif sososssssssssssooss or sooosssssssssssooss, soooossssssssssooss or sssssssssssssssssss, in order from 5’ to 3', where each “s?’ represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester internucleoside linkage. In certain embodiments, each “s” represents a (Sp)-phosphorothioate internucleoside linkage.

[0516] In certain embodiments, an oligomeric compound comprises a modified oligonucleotide consisting of 18-22 linked nucleosides, 19-21 linked nucleosides or 20 linked nucleosides, where the modified oligonucleotide comprises includes at least one modification selected from a modified sugar moiety, or a modified PATENT 7328*158807PCT / NLM-1005-PC internucleoside linkage, or a modified sugar moiety and a modified internucleoside linkage, and die modified oligonucleotide consists of a nucleobase sequence complementary to: (i) an equal length portion of nucleobases 3458-3708, 3624-3727 or 3644-3707 of SEQ ID NO: 1; or (ii) an equal length portion of nucleobases 3644-3663, 3647-3666, 3652-3671, 3663-3682, 3666-3685, 3677-3696 or 3688-3707 of SEQ ID NO:1. In certain embodiments, the modified oligonucleotide comprises or consists of at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of a nucleobase sequence selected from: SEQ ID NO:4 to SEQ ID NO:7 and SEQ ID NO:9 to SEQ ID NO: 11. In certain embodiments, the modified oligonucleotide is a gapmer that includes a 5'~region, a central region and a 3’-region. In certain embodiments, the modified oligonucleotide is a MOE gapmer. In certain embodiments, the modified oligonucleotide is a 5-10-5 MOE gapmer. In certain embodiments, the sugar motif for the gapmer is (from 5’ to 3’) eeeeeddddddddddeeeee, where each “d” represents a 2’ -beta-D-deoxy ribosyl sugar moiety, and each “e” represents a 2’- MOE sugar moiety. In certain embodiments, the internucleoside linkages of the modified oligonucleotide are phosphorothioate internucleoside linkages and phosphodiester internucleoside linkages. In certain embodiments, the modified oligonucleotide comprises or consists of internucleoside linkages according to the linkage motif sososssssssssssooss, sooosssssssssssooss, soooossssssssssooss or sssssssssssssssssss, in order from 5' to 3’, where each “s” represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester internucleoside linkage. In certain embodiments, each “s” represents a (Sp)-phosphorothioate internucleoside linkage.

[0517] In certain embodiments, an oligomeric compound comprises a modified oligonucleotide consisting of 18-22 linked nucleosides, 19-21 linked nucleosides or 20 linked nucleosides, where the modified oligonucleotide comprises includes at least one modification selected from a modified sugar moiety, or a modified internucleoside linkage, or a modified sugar moiety and a modified intemucleoside linkage, and the modified oligonucleotide consists of a nucleobase sequence complementary to: an equal length portion of nucleobases 3643-3702, 3653-3692 or 3663-3682 of SEQ ID NO:1. In certain embodiments, the modified oligonucleotide comprises or consists of at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases PATENT 7328*158807PCT / NLM-1005-PC of a nucleobase sequence of SEQ ID NO:4. In certain embodiments, the modified oligonucleotide is a gapmer that includes a 5 ’-region, a central region and a 3 ’-region. In certain embodiments, the modified oligonucleotide is a MOE gapmer. In certain embodiments, the modified oligonucleotide is a 5-10-5 MOE gapmer. In certain embodiments, the sugar motif for the gapmer is (from 5’ to 3’) eeeeeddddddddddeeeee, where each “d” represents a 2' -beta-D-deoxy ribosyl sugar moiety, and each “e” represents a 2’- MOE sugar moiety. In certain embodiments, the internucleoside linkages of the modified oligonucleotide are phosphorothioate internucleoside linkages and phosphodiester internucleoside linkages. In certain embodiments, the modified oligonucleotide comprises or consists of internucleoside linkages according to the linkage motif sososssssssssssooss, sooosssssssssssooss, soooossssssssssooss or sssssssssssssssssss, in order from 5’ to 3’, where each "‘s'’ represents a phosphorothioate internucleoside linkage and each “o’’ represents a phosphodi ester internucleoside linkage. In certain embodiments, each “s” represents a (Sp)-phosphorothioate internucleoside linkage.

[0518] In certain embodiments, an oligomeric compound comprises a modified oligonucleotide consisting of 18-22 linked nucleosides, 19-21 linked nucleosides or 20 linked nucleosides, where the modified oligonucleotide comprises includes at least one modification selected from a modified sugar moiety, or a modified internucleoside linkage, or a modified sugar moiety and a modified internucleoside linkage, and the modified oligonucleotide consists of a nucleobase sequence complementary to an equal length portion of nucleobases 720-3074, 2908-2967 or 2928-2947 of SEQ ID NO:1. In certain embodiments, the modified oligonucleotide comprises or consists of at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of a nucleobase sequence selected from: SEQ ID NO: 12. In certain embodiments, the modified oligonucleotide is a gapmer that includes a 5’-region, a central region and a 3’-region. In certain embodiments, the modified oligonucleotide is a MOE gapmer. In certain embodiments, the modified oligonucleotide is a 5-10-5 MOE gapmer. In certain embodiments, the sugar motif for the gapmer is (from 5’ to 3’) eeeeeddddddddddeeeee, where each “d” represents a 2' -beta-D-deoxy ribosyl sugar moiety, and each “e” represents a 2’- MOE sugar moiety. In certain embodiments, the internucleoside linkages of the modified oligonucleotide are phosphorothioate PATENT 7328*158807PCT / NLM-1005-PC internucleoside linkages and phosphodiester internucleoside linkages. In certain embodiments, the modified oligonucleotide comprises or consists of internucleoside linkages according to the linkage motif sososssssssssssooss, sooosssssssssssooss, soooossssssssssooss or sssssssssssssssssss, in order from 5’ to 3’, where each “s” represents a phosphorothioate internucleoside linkage and each ‘"o” represents a phosphodiester internucleoside linkage. In certain embodiments, each “s” represents a (Sp)-phosphorothioate internucleoside linkage.

[0519] In certain embodiments, an oligomeric compound comprises a modified oligonucleotide consisting of 18-22 linked nucleosides, 19-21 linked nucleosides or 20 linked nucleosides, where the modified oligonucleotide comprises includes at least one modification selected from a modified sugar moiety, or a modified internucleoside linkage, or a modified sugar moiety and a modified internucleoside linkage, and the modified oligonucleotide consists of a nucleobase sequence complementary to an equal length portion of nucleobases 720-2935, 2200-2259 or 2220-2239 of SEQ ID NO:1. In certain embodiments, the modified oligonucleotide comprises or consists of at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of a nucleobase sequence selected from: SEQ ID NO:8. In certain embodiments, die modified oligonucleotide is a gapmer that includes a 5’-region, a central region and a 3’-region. In certain embodiments, the modified oligonucleotide is a MOE gapmer. In certain embodiments, the modified oligonucleotide is a 5-10-5 MOE gapmer. In certain embodiments, the sugar motif for die gapmer is (from 5’ to 3’) eeeeeddddddddddeeeee, where each “d'” represents a.2' -beta-D-deoxy ribosyl sugar moiety, and each “e” represents a 2’- MOE sugar moiety. In certain embodiments, the internucleoside linkages of the modifi ed oligonucleotide are phosphorothioate internucleoside linkages and phosphodiester internucleoside linkages. In certain embodiments, the modified oligonucleotide comprises or consists of internucleoside linkages according to the linkage motif sososssssssssssooss, sooosssssssssssooss, soooossssssssssooss or sssssssssssssssssss, in order from 5’ to 3’, where each “s” represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester intemudeoside linkage. In certain embodiments, each “s” represents a (Sp)-phosphorothioate internucleoside linkage. PATENT 7328*158807PCT / NLM-1005-PC In certain embodiments, an oligomeric compound is not allele selective, and does not inhibit expression of a first SAA1 allele significantly more than a second SAA1 allele. Allele selectivity can be assessed by a selectivity ratio, and a selectivity ratio of less than 2 typically is indicative that an oligomeric compound is not allele selective. In certain embodiments, a selectivity ratio is determined according to inhibition of expression of a first SAA1 allele and inhibition of expression of a second SAA1 allele in response to an oligomeric compound. In certain embodiments, the first SAA1 allele is a mutant SAA1 allele containing a pathogenic genetic variant (the genetic variant c.101A>T (resulting in p. Asp34Val) for example). In certain embodiments, the second SAA1 allele is a wild-type SAA1 allele not containing the pathogenic genetic variant. In certain embodiments, a selectivity ratio is an IC50 value determined for the second SAA1 allele (numerator) to an IC50 value determined for the first SAA1 allele (denominator), with each IC50 value determined by a multi-point dose response curve. In certain embodiments, such a selectivity ratio is less than 2 for an oligomeric compound that is not allele selective.

[0520] In certain embodiments, an oligomeric compound oligonucleotide comprises at least one methyl uracil (mU) nucleoside. In certain embodiments, an oligomeric compound oligonucleotide comprises at least one mU nucleoside and at least one thymidine nucleoside. In certain embodiments, an oligomeric compound oligonucleotide contains two, or at least two, mU nucleosides, optionally within a terminal 3’ portion of the oligonucleotide (a terminal 3’ portion of the oligonucleotide containing the last five nucleosides of the oligonucleotide for example). In certain embodiments, an oligonucleotide contains two, or at least two, mU nucleosides and at least one thymidine nucleoside. In certain embodiments, an oligomeric compound oligonucleotide consists of 20 linked nucleosides. In certain embodiments, an oligomeric compound is not according to the formula:

[0521] GalNAcoAmCmCmCATTGTGTAmCmCmCTmCTTmCmC (SEQ ID NO: 17)

[0522] where AmCmCmCATTGTGTAmCmCmCTmCTTmCmC (SEQ ID NO: 18) is an oligonucleotide, each mC is a methyl cytidine oligonucleotide, GalNAc is a conjugate group containing multiple N-acetylgalactosamine moieties, and “o” is a phosphodiester moiety. In certain embodiments, each internucleoside linkage of the oligonucleotide AmCmCmCATTGTGTAmCmCmCTmCTTmCmC (SEQ ID NO: 18) PATENT 7328*158807PCT / NLM-1005-PC is a phosphorothioate internucleoside linkage. In certain embodiments, an oligomeric compound oligonucleotide does not consist of the nucleobase sequence ACCCATTGTGTACCCTCTTCC (SEQ ID NO: 19). In certain embodiments, an oligomeric compound oligonucleotide does not consist of 21 linked nucleosides. Exemplary Target Nucleic Acids in Certain Tissues

[0523] In certain embodiments, oligomeric compounds comprise or consist of an oligonucleotide comprising a region that is complementary to a target nucleic acid, where the target nucleic acid is expressed in a pharmacologically relevant tissue. In certain embodiments, a pharmacologically relevant tissue includes a liver cell, and optionally a hepatocyte cell. In certain embodiments, a pharmaceutically relevant tissue includes a liver cell (a hepatocyte cell for example) or other cell to which an oligomeric compound can be targeted.

[0524] Exemplary Methods and Uses

[0525] Certain embodiments provided relate to methods of reducing or inhibiting SAA1 expression or activity, which can be useful for treating, preventing, or ameliorating a SAA1-associated medical condition. In certain embodiments, the SAA1 -associated medical condition is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure and arrhythmia. Amyloid A amyloidosis (AA amyloidosis) is a serious clinical condition resulting from the systemic deposition of amyloid A originating from serum amyloid A proteins with the kidneys being the most commonly and earliest affected organ. AA amyloidosis typically occurs as a reaction to another illness, such as a chronic inflammatory disease or a chronic infection. Infections and inflammation can cause the liver to produce a protein called SAA (serum amyloid A protein) at high levels. The first symptom of AA amyloidosis is usually protein in the urine. Often proteinuria (protein in the urine) becomes massive, and nephrotic syndrome develops. Patients can lose so much protein in their urine that they develop low levels of protein (albumin) in their blood, leading to swelling of the ankles and legs (edema). A high cholesterol level also can present with this syndrome. As a result, patients can develop renal failure and the need for dialysis. AA amyloidosis can involve other organs in addition to the kidneys. An enlarged spleen, enlarged liver, and enlarged thyroid are not uncommon. Autonomic neuropathy is frequent with symptoms of orthostatic hypotension (low blood pressure on standing), gastrointestinal atony (slowing of stomach emptying) PATENT 7328*158807PCT / NLM-1005-PC and diarrhea or constipation. AA amyloid deposits in the heart causing congestive heart failure and arrhythmias (irregular heart beat) may develop later in the course of the disease.

[0526] In certain embodiments, a method comprises administering to a subject an oligomeric compound comprising a modified oligonucleotide having a nucleobase sequence complementary to an equal length portion of SAA1 nucleic acid, or a composition containing the compound as described herein. In certain embodiments, a method for treating a SAA1 -associated medical condition comprises administering to a subject an oligomeric compound comprising a modified oligonucleotide having a nucleobase sequence complementary to an equal length portion of a SAA1 nucleic acid, or a composition containing the compound as described herein. In certain embodiments, the subject has an elevated SAA1 level. In certain embodiments, a method of reducing expression of SAA1, for example, reducing expression of SAA1 RNA, and / or reducing expression of SAA1 protein in a cell comprises contacting the cell with an oligomeric compound comprising a modified oligonucleotide having a nucleobase sequence complementary to an equal length portion of SAA1 nucleic acid, or a composition containing the compound as described herein. In certain embodiments, the cell has an elevated SAA1 level. Certain embodiments are drawn to an oligomeric compound comprising a modified oligonucleotide having a nucleobase sequence complementary to an equal length portion of SAA1 nucleic acid, or a composition containing the compound as described herein, for use in treating SAA1-associated medical condition, such as amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia, for example. In certain embodiments, the SAA1 -associated medical condition is associated with an elevated SAA1 level. An oligomeric compound utilized can be any suitable oligomeric compound described herein.

[0527] In certain embodiments, an elevated SAA1 level is an elevated level of a SAA1 nucleic acid (SAA1 RNA, SAA1 pre-mRNA and / or SAA1 mRNA, for example) and / or SAAI protein. An elevated SAAI level often is relative to a control SAA1 level of an SAA1 nucleic acid or SAA1 protein, and in certain embodiments, a control level is of a SAA1 nucleic acid and / or SAA1 protein that does not contain a pathogenic variant (a pathogenic genetic variant for example). In certain embodiments, an oligomeric compound is administered in an amount sufficient to PATENT 7328*158807PCT / NLM-1005-PC decrease an elevated SAA1 level. In certain embodiments, an elevated SAA1 level is reduced by at least 20%, by at least 30%, by at least 40%, by at least 50%, by at least 60%, by at least 70%, by at least 80%, by at least 90%, or by at least 95%, after administration of an oligomeric compound. In certain embodiments, an elevated SAA1 level is in blood, such as serum or plasma for example. A SAA1 level can be identified using any suitable method, including without limitation a quantitative polymerase chain reaction method (a RT-qPCR method as described in Example 1 for example) or an immunologic assessment process (enzyme linked immunosorbent assay (ELISA), immunonephelometry assay (immunonephelometry light scattering assay or latex-enhanced immunonephelometry assay for example) or Western blot, for example). In certain embodiments, a sample, cell, tissue and / or subject has an elevated SAA1 level and a SAA1 nucleic acid therein includes a pathogenic genetic variant or does not include a pathogenic genetic variant. A pathogenic genetic variant can be a variant described herein, such as a pathogenic genetic variant in exon 3 of a SAA1 nucleic acid, a pathogenic genetic variant in a SAA1 nucleic acid resulting in a Asp34Val modification of a SAA1 protein, and / or a c.101 A> T pathogenic genetic variant in a SAA1 nucleic acid. In certain embodiments, a subject has or is at risk for developing a SAA1 -associated medical condition. In certain embodiments, the subject has a SAA1 -associated medical condition and in certain instances has been diagnosed as having a SAA1 -associated medical condition. In certain embodiments, at least one symptom or hallmark of a SAA1 -associated medical condition is ameliorated. In certain embodiments, a SAA1-associated medical condition is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia. In certain embodiments, a symptom or hallmark is one or more of proteinuria, nephrotic syndrome, low blood protein level, low blood albumin level, edema, high cholesterol, enlarged spleen, enlarged liver, enlarged thyroid, orthostatic hypotension, gastrointestinal atony, diarrhea and constipation.

[0528] Certain embodiments include determining presence or absence of a pathogenic genetic variant in a SAA1 nucleic acid. Certain embodiments include determining a genotype of a pathogenic genetic variation site and / or position in a SAA1 nucleic acid. In certain embodiments, presence or absence of a non-loss of function genetic variant is determined, which in certain instances can be a dominant negative genetic variant or gain of function genetic variant. Certain embodiments include determining PATENT 7328*158807PCT / NLM-1005-PC presence or absence of a pathogenic genetic variant (determining a genotype for example) resulting in an amino acid modification (an amino acid substitution, for example) in a SAA1 protein selected from: p. Leu3Pro, p. Val8Ala, p. Asp34Val, p. Ala70Val, p. Trp71Arg, p. Val75A1a, p. Asp93Glu, p. Gly90Asp, p. Asn 101 Asp or p. Lys121Arg. Certain embodiments include determining presence or absence of a pathogenic genetic variant (determining a genotype for example) at a single nucleotide variation site chosen from: rs4638289, rs111154228, rs1829575, rs11545470, rs11545474, rs1136743, rs11545468, rs1136747, rs15790, rs79681911, rs12218, rs17850140 or rs61745680. Certain embodiments include determining presence or absence of a pathogenic genetic variant (determining a genotype for example) in a SAA1 nucleic acid selected from: c.8T>C, c.23T>C, c.209C>T, c.101A>T, c.211T> C, c.224T> C, c.279C>T, c.269G> A, c.288T> C, c.301A> G or c.362A>C, Certain embodiments include determining presence or absence of a pathogenic genetic variant (determining a genotype for example) resulting in an amino acid substitution in a SAA1 protein selected from: p. Glu74Asp (resulting from c.222A> C for example), p. Phe86Leu (resulting from c.256T> C for example), p. Phe87Ile (resulting from c.259T> A for example): p. Phe87Ser (resulting from c 260T> C for example); p. Phe87Leu (resulting from c 261T> A for example), р. Ala99Gly (resulting from c.296C> G for example); p. TrplO3Cys (resulting from с.309G> C for example), p. Lys105fs (“fs” is a “frame shifty; resulting from c.323_324del for example); p. Asn111Ser (resulting from c.332A> G for example) or p. Arg114Pro (resulting from c.341G> C for example).

[0529] Certain embodiments include determining presence or absence of a pathogenic genetic variant in an exon, optionally in exon 3 of a SAA1 nucleic acid (determining a suitable genotype for example). Certain embodiments include determining presence or absence of a pathogenic genetic variant in a SAA1 nucleic acid resulting in a Asp34Val modification of a SAA1 protein (determining a suitable genotype for example). In certain embodiments, presence or absence of a c.101A>T pathogenic genetic variant is determined in a SAA1 nucleic acid (a suitable genotype is determined for example). Certain embodiments include administering an oligomeric compound targeted to a SAA1 nucleic acid if presence of a pathogenic genetic variant is determined (for example, if a pathogenic genetic variant genotype is identified). PATENT 7328*158807PCT / NLM-1005-PC A pathogenic genetic variation site and / or a pathogenic genetic variation position can be characterized for a SAA1 nucleic acid (for example, a genotype can be determined) in situ or from a sample. A sample can be a sample containing cells or tissue, and can be from a subject. A sample from a subject can be a solid sample or liquid sample.

[0530] Presence or absence of a pathogenic genetic variant, and / or a suitable genotype, can be determined in any suitable manner known, including by sequencing SAA1 nucleic acid and / or by utilizing nucleic acid probes targeted to a genetic variation site, for example. Any suitable sequencing process can be utilized, including a sequencing process that determines sequences from long reads, or from short reads, or from short reads and long reads for example. In certain embodiments, short read sequencing processes typically generate sequencing reads of about 50 to about 300 nucleobases often by sequencing by synthesis or ligation processes. Non-limiting examples of short read sequencing processes include (i) solid phase bridge amplification followed by labeled nucleotide incorporating monitored by fluorescence detection (Illumina); (ii) emulsion PCR, generating microbead-bound DNA clones, followed by nucleotide incorporation monitored by pyrophosphate release (454 pyrosequencing); (iii) emulsion PCR, generating microbead-bound DNA clones, followed by nucleotide incorporation monitored by a pH sensor as protons are released (Ion Torrent); (iv) emulsion PCR followed by hybridization to amplicons a complementary strand grown by DNA ligase (SOLID); and (v) hybridizing an anchor sequence and probes to DNA template in a series of ligation reactions taking place on a nanoball (cPAL). In certain embodiments, a long read sequencing process generates nucleobase sequence reads of about 1,000 to about 30,000 nucleobases. Non-limiting examples of long read sequencing processes include nanopore sequencing processes (Oxford Nanopore Technologies) and zero-mode waveguide processes (Pacific Biosciences single molecule real-time (SMRT) sequencing). Certain embodiments include determining presence or absence of an elevated level of a SAA1 nucleic acid and / or SAA1 protein. An elevated level of a SAA1 nucleic acid or SAA1 protein often is relative to a control level of an SAA1 nucleic acid or SAA1 protein. In certain embodiments, a control level is in cells in which SAA1 nucleic acid does not contain a pathogenic genetic variant. In certain embodiments, an oligomeric compound targeted to a SAA1 nucleic acid is administered if an elevated level of a SAA1 nucleic PATENT 7328*158807PCT / NLM-1005-PC acid and / or SAA1 protein is present, detected and / or observed. In certain embodiments, (i) presence, absence and / or amount of an elevated level of a SAA1 nucleic acid and / or SA A 1 protein is determined, and (ii) an oligomeric compound targeted to a SAA1 nucleic acid is administered if an elevated level of a SAA1 nucleic acid and / or SAA1 protein is present. In certain embodiments, presence, absence and / or amount of an elevated level of a SAA1 nucleic acid and / or SAA1 protein is determined for a sample, cell(s), tissue and / or subject. In certain embodiments, an oligomeric compound targeted to a SAA1 nucleic acid is administered to a cell, cells, a tissue and / or a subject. In certain embodiments, a SAA1 nucleic acid is a SAA1 RNA, such as pre-mR A and / or mRNA for example. Presence, absence and / or amount of an elevated level of a SAA1 nucleic acid and / or SAA1 protein can be determined using any suitable method, including without limitation, a quantitative polymerase chain reaction (qPCR) process (as described in Example I herein for example) for SAA1 nucleic acid, or by an immunologic assessment method (Western blot analysis, or enzyme-linked immunosorbent assay (ELISA), or immunonephelometry assay (immunonephelometry light scattering assay or latex-enhanced immunonephelometry assay for example), for example) for SAA1 protein, for example.

[0531] Exemplary Pharmaceutical Compositions

[0532] In certain embodiments, provided is a pharmaceutical composition comprising one or more oligomeric compounds described herein. In certain embodiments, the one or more oligomeric compounds each consists of a modified oligonucleotide. In certain embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable diluent or carrier. In certain embodiments, a pharmaceutical composition compri ses or consists of a sterile saline solution and one or more oligomeric compounds. In certain embodiments, the sterile saline is pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric compound and sterile water. In certain embodiments, the sterile water is pharmaceutical grade water. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric compounds and phosphate-buffered saline (PBS). In certain embodiments, the sterile PBS is pharmaceutical grade PBS. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric compounds and artificial PATENT 7328*158807PCT / NLM-1005-PC cerebrospinal fluid (“artificial CSF' or “aCSF'’). In certain embodiments, the artificial cerebrospinal fluid is pharmaceutical grade.

[0533] In certain embodiments, a pharmaceutical composition comprises a modified oligonucleotide and artificial cerebrospinal fluid (aCSF). In certain embodiments, a pharmaceutical composition consists of a modified oligonucleotide and artificial cerebrospinal fluid. In certain embodiments, a pharmaceutical composition consists essentially of a modified oligonucleotide and artificial cerebrospinal fluid. A pharmaceutical composition consisting essentially of a modified oligonucleotide and artificial cerebrospinal fluid can include one or more other components that do not. materially affect solubility of the modified oligonucleotide in the pharmaceutical composition. In certain embodiments, the artificial cerebrospinal fluid is pharmaceutical grade.

[0534] In certain embodiments, aCSF comprises sodium chloride, potassium chloride, sodium dihydrogen phosphate dihydrate, sodium phosphate dibasic anhydrous, calcium chloride dihydrate, and magnesium chloride hexahydrate. In certain embodiments, the pH of an aCSF solution is modulated with a suitable pH-adjusting agent, for example, with acids such as hydrochloric acid and alkalis such as sodium hydroxide, to a range of from about 7.1-7.3, or to about 7.2.

[0535] In certain embodiments, a pharmaceutical composition comprises one or more oligomeric compounds and one or more excipients. In certain embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

[0536] In certain embodiments, an oligomeric compound can be admixed with pharmaceutically acceptable active and / or inert substances for the preparation of a pharmaceutical composition or formulation. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0537] In certain embodiments, an oligomeric compound in a pharmaceutical composition can include any pharmaceutically acceptable salt of the oligomeric compound, ester of the oligomeric compound, or salt of such ester. In certain embodiments, a pharmaceutical composition comprising an oligomeric compound, PATENT 7328*158807PCT / NLM-1005-PC upon administration to a subject, including a human subject, is capable of providing (directly or indirectly) a biologically active metabolite or residue of the oligomeric compound. For example, included are pharmaceutically acceptable salts of oligomeric compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. In certain embodiments, pharmaceutically acceptable salts comprise inorganic salts, such as monovalent or divalent inorganic salts. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium, potassium, calcium, and magnesium salts. In certain embodiments, prodrugs comprise one or more conjugate group attached to an oligonucleotide, where the conjugate group is cleaved by endogenous nucleases within a subject. In certain embodiments, oligomeric compounds are lyophilized and isolated as sodium salts. In certain embodiments, a sodium salt of an oligomeric compound is mixed with a pharmaceutically acceptable diluent. In certain embodiments, a pharmaceutically acceptable diluent comprises sterile saline, sterile water, PBS, or aCSF. In certain embodiments, a sodium salt of an oligomeric compound is mixed with PBS. In certain embodiments, a sodium salt of an oligomeric compound is mixed with aCSF.

[0538] Lipid moieties have been used in nucleic acid therapies in a variety of methods. In certain methods, an oligomeric compound is introduced into preformed liposomes or lipoplexes comprising mixtures of cationic lipids and neutral lipids. In certain methods, complexes containing oligomeric compound with mono- or polycationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to a particular cell or tissue. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to fat tissue. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to muscle tissue.

[0539] In certain embodiments, a pharmaceutical composition comprises a delivery system. Non-limiting examples of delivery’ systems include liposomes and emulsions. Certain delivery systems are useful for preparing certain pharmaceutical compositions including those comprising hydrophobic compounds. In certain embodiments, certain organic solvents such as dimethylsulfoxide are used.

[0540] In certain embodiments, a pharmaceutical composition comprises one or more tissuespecific delivery molecules designed to deliver an oligomeric compound to specific PATENT 7328*158807PCT / NLM-1005-PC tissues or cell types. For example, in certain embodiments, a pharmaceutical composition includes liposomes coated with a tissue-specific antibody.

[0541] In certain embodiments, a pharmaceutical composition comprises a co-solvent system. Certain co-solvent systems comprise, for example, benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w / v benzyl alcohol, 8% w / v of the nonpolar surfactant Polysorbate 80™ and 65% w / v polyethylene glycol 300. Proportions of components in a co-solvent system can be varied without significantly altering their solubility and toxicity characteristics. Components of a co-solvent system can be vari ed: for example, other surfactants can be used instead of Polysorbate 80™; the fraction size of polyethylene glycol can be varied; other biocompatible polymers may replace polyethylene glycol (with polyvinyl pyrrolidone for example); and other sugars or polysaccharides may substitute for dextrose.

[0542] In certain embodiments, a pharmaceutical composition is prepared for oral administration. In certain embodiments, a pharmaceutical composition is prepared for buccal administration. In certain embodiments, a pharmaceutical composition is prepared for systemic administration, and optionally for subcutaneous administration. In certain embodiments, a pharmaceutical composition is prepared for administration by injection (for example, intravenous, subcutaneous, intramuscular, intrathecal (IT), intracerebroventricular (ICV) and the like). In certain embodiments, a pharmaceutical composition comprises a carrier and is formulated in an aqueous solution, such as water or a physiologically compatible buffer such as Hanks’s solution, Ringer's solution, or physiological saline buffer. In certain embodiments, oner or more other ingredients are included, including one or more ingredients that aid in solubility or serve as preservatives for example. In certain embodiments, injectable suspensions are prepared using appropriate liquid carriers, suspending agents and the like. In certain embodiments, a pharmaceutical composition for injection is provided in unit dosage form, and can be provided in an ampoule or in a multi -dose container for example. In certain embodiments, a pharmaceutical composition for injection is a suspension, solution or emulsion in an oily or aqueous vehicle, and optionally can contain a fonnulatory agent such as a suspending, stabilizing and / or dispersing agent. Nonlimiting examples of solvents suitable for use in pharmaceutical compositions for PATENT 7328*158807PCT / NLM-1005-PC injection include lipophilic solvents and fatty oils (sesame oil for example), synthetic fatty acid esters (ethyl oleate or triglycerides for example), and liposomes.

[0543] Under certain conditions, an oligomeric compound can function as an acid. While an oligomeric compound can be depicted or described in protonated (free acid) form or ionized and in association with a cation (salt) form, aqueous solutions of such compounds exist in equilibrium among such forms. For example, a phosphodiester linkage of an oligonucleotide in aqueous solution typically exists in equilibrium among free acid, anion and salt forms. In certain embodiments, a composition comprising a plurality of oligomeric compound molecules includes all such forms. Also, for example, certain oligonucleotides have several linkages, each of which is in equilibrium, and oligonucleotides in solution typically exist in an ensemble of forms at multiple positions all at equilibrium. In certain embodiments, a composition comprising a plurality of oligomeric compound molecules includes all such forms. Depicted structures necessarily depict a single form, and unless otherwise indicated, such depictions include corresponding forms. For example, a structure depicting the free acid of an oligomeric compound followed by the term ‘for a pharmaceutically acceptable salt thereof” Includes all such forms that can be fully or partially protonated, de-protonated and in association with a cation or a combination of cations. In certain embodiments, one or more specific cations is identified. Non-limiting examples of cations include sodium, potassium, calcium, and magnesium. In certain embodiments, a structure depicting the free acid of a compound followed by the term “or a pharmaceutically acceptable salt thereof' includes all such forms that can be fully or partially protonated, de-protonated and in association with one or more cations selected from sodium, potassium, calcium, and magnesium.

[0544] In certain embodiments, an oligomeric compound is in aqueous solution with sodium. In certain embodiments, an oligomeric compound is in aqueous solution with potassium. In certain embodiments, an oligomeric compound is in PBS. In certain embodiments, an oligomeric compound is in water. In certain embodiments, the pH of the solution is adjusted with a base (NaOH for example) and / or an acid (HCl for example) to achieve a desired pH.

[0545] An oligomeric compound can be provided in a pharmaceutical composition at a suitable dose. A dose can be in the form of a dosage unit. A dose or dosage unit of a modified oligonucleotide or an oligomeric compound in milligrams typically indicates PATENT 7328*158807PCT / NLM-1005-PC the mass of the free acid form of the modified oligonucleotide or oligomeric compound. While the free acid is in equilibrium with anionic and salt forms as described, the dose is calculated with an assumption that the modified oligonucleotide or oligomeric compound exists as a solvent-free, sodium-acetate free, anhydrous, free acid.

[0546] In certain embodiments, where a modified oligonucleotide or an oligomeric compound is in a solution comprising sodium (for example, saline), the modified oligonucleotide or oligomeric compound can be partially or fully deprotonated and in association with sodium ions. The mass of the protons typically is counted toward the weight of the dose, and the mass of the sodium ions typically is not counted toward the weight of the dose.

[0547] In certain embodiments, where a modified oligonucleotide or oligomeric compound is in a solution, such as aCSF, comprising sodium, potassium, calcium, and magnesium, the modified oligonucleotide or oligomeric compound can be partially or fully de-protonated and in association with sodium, potassium, calcium, and / or magnesium. The mass of the protons typically is counted toward the weight of the dose, and the mass of the sodium, potassium, calcium, and magnesium ions typically is not counted toward the weight of the dose.

[0548] In certain embodiments, where an oligomeric compound comprises a conjugate group, the mass of the conjugate group can be included in calculating the dose of such oligomeric compound. If the conjugate group also has an acid, the conjugate group typically is assumed as being fully protonated for the purpose of calculating dose.

[0549] Products of manufacture and Kits

[0550] Provided are products of manufacture and kits for practicing methods as provided herein. Optionally, products of manufacture and kits further comprise instructions for practicing methods as provided herein.

[0551] Exemplary, sequences

[0552] SEQ ID NO:1

[0553] > 11 dna: chromosome: GRCh38:11:18266260:18269977:1 AGGCAGGGACCCGCAGCTCAGCTACAGCACAGATCAGGTGAGGAGCACA CCAAGGAGTGATTTTTAAAACTTACTCTGTTTTCTCTTTCCCAACAAGATT ATCATTTCCTTTAAAAAAAATAGTTATCCTGGGGCATACAGCCATACCATT PATENT 7328*158807PCT / NLM-1005-PC CTGAAGGTGTCTTATCTCCTCTGATCTAGAGAGGTAAGCAGGGTCGGGCC TGGTAGTACTTGGATGGGAGAACACCTGGGAATACCAGGTGCTAAAGGCT TTAAGAATAAAAAATAATGATCCTGCTTTGTGTTTATCCCATGTTGAGTTC TGTGCGGGGCAGAGGGAACACACGGTAAATGCGTTATGGGGAATTATAG GCTACTTGAGGGAGTGACAGTCTGGTGGTAACTCCTGCCTTCCTCCATCAG TGCCACGTTGGCATCCTCTTATGCAGTCAGGCTTCAGGGCTGATGGGTTCA GAACCGAGGGCTTCTGGCTCTGAGTGAGGTCCTGCTGCAAGGTTTCCTAG ATGAGCCACTGAGACTCTAATAAGATCCAGTGGAAATAACCAGGCTCTCG TCGGAATATAAGTCCCAAGGGAAGCTGTGCCAGTCTTGTGGGCGACTGCC TGACTTCTCCTTTCATTTCAGCACCATGAAGCTTCTCACGGGCCTGGTTTTC TGCTCCTTGGTCCTGGGTGTCAGCAGCCGAAGCTTCTTTTCGTTCCTTGGC GAGGCTTTTGATGGTAAGGCTTCAGAAGGTTTGCAGGATTTCTGAAGAGA AACATCACCCTGGACCTGATAAACTGGGGAAAATGATGCTTTCGGAAGGC TGCTTTTGAACCACAGAGTTGCTAGTGTCTGCGTTGCTGAGGCCTGCCAGG AACTAGGGTTTGCTGGGTTGCCTGTCTCGAGTCTTTCAGAGCTGCTGGGAA TATCCCCTTTCCCCGTAGTGCAGCTTCTCAGGATGTGTTAAGTGGATGGAT CACATTTCAGAAGCCGCTGCAAGGTGTATCAAAAACACATCTCCTGAGCC GTAAGGGACGGGGCATCCAGTAACAACGCACACGGGGTATTTTTGGGCTT CCTTAAGATTTGAGCCGCTGCCTTAGGTTGTGCTGCCCAATGTGCCTGGGG AGCTGCTAAACAGATTAGAGAGTCGAGGATTGTTGTCAGTTACTCAGAGA AAGAACAATCATCCTTTCCAGGAGCACCTGAGCTGTTTGTTTTGCGTAGAA GATGCAAAATAAGGCCTGCAATGGGTATAAAATGTCCCTCAGCATAAATC GCATAGGAGTATGACTAAGGCTGTTGACTCTTCTGTCTTCTTTCTCCTTCCT CCTTCGATTTCCTAGTTGGATAATGTACAGGGCTCTTTAGCCTCGCTCTGT CAGGGGCTCCCTTCCTGGTTTGTTCTGTTTCCATTCTTCCTTCTCCAGCCTT CTTGACAAGAGCTGGGAACTAACGTGCCTCAAGCCCCCACAAGGACCACA GCATTTTCTCATTTAGTTTCAGAATGACTCTGTGACGCAATCTTCCTCTCTT GGAAGGTGAGAAAGCTGATCTTGGAAGGTGAGAAAGCTGAGACTTAGAG CAGCTGAAGCCAATGCCCAGGGACTTACTGCCAGTCAGCAGGTGGCAGGG CAGAGGTTTGAGCCCGGCTGTGCTTGAGGTCAGGGCTCTTGCCAGGTAGA CGCATCACTGACCACCTCCTAGAGGTTGATGGTTATGAATCTCAGGCACA CCTTGGCATCACCTGAAATACCCATGCCTTCAACTCCCCAGCAGAGTCTGC AGAAACTGGCCTGGGGTGTGGCCTGGGCACTGGGACTTTCAGTTTCTCTCT GGGTGATTAGAAAGTGCAGCCAAGGCTCACGCCTGTAATTCCAGCACTTT GGGAGGCCAAGGTGGATGAATCACTTGAGGTCATGAGTTCCGGAGCAGCC TGGCCAACATGGTGAAACCCCGTCTCTACTAAAAATACTAAAATGTAGCC AGGCGTGGTGGCAGGCACCTGTAATCCCAGCTACTCAGGAGGCTGAAGCA CGAGAATCACTTGAACCCGAGAAGCAGAGGTTGCAGTGACTAGAGATCG CACCAGTGTCCTCCAACCTGGGTGACAGAGCGAGACTCCATCTAAAAAAA ATGAAAAAGAAAGTGCAGCCAAGGCAGAGCACCACTGCCCTATTGCTTCC TCAAGCAACCCACAGCATCAGTACAGCCTACTAAGAAAGTATTTAGGGAC TTTTATGCTCCTAACAGTCACTGGAACTCACGTCACAATGACGTGTATTCC ATTTGCAAGAATATATACTTTAGGTCGGGGTGCGGTGGCTCACGCCTGTA ATCCCAGCACTTTGGGAGGCCAAGGCAGGGGGATCACGAGGTCAGGAGT TCGAGACCAGCCTGACCAACATGGTGAAATCCCCGTCTCTACTAAAAATA CAAAAATTAGCCAGGCGTGATGGCGCATGCCTGTAATCTCAGCTACTCAG GAGGCTGAGGCAGAAGAATCTCTTGAACCTGGGAGGTGGAGGTTGCGAT GAGCTGAGATAGCACCACTGCACTCCAGCCTGGGCGACAGAGCAAGACTC TGTCTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGAATATAAACTT TAGTAGTCAGGGCAGAAGTACTCTGTGTCTGCCACCTTTCTCAGCATCAGT PATENT 7328*158807PCT / NLM-1005-PC ATTCCATGTCACTACCTCATTCATACACACTCCTGGATCTTATCATAGGCA GCTTCATTCTATAGCAGTGGCTCTTCACCAGGGCACTTGAAGAAGCCAAC TAGGATAAAGGAATGTGCTTCTCAACCCATGGTATCCAAGGCTGCTATGA TCACAGGCTGAAAGCTTGAAGTCAGTGGAAGATTTGTCCTTCCTCATTCCC CTCTAAGGTGTTGTTGGAGTCTTTATGTTCTCCTGATGTCCCTTCTGCCTTT CCTTTCCTTTCCAGGGGCTCGGGACATGTGGAGAGCCTACTCTGACATGA GAGAAGCCAATTACATCGGCTCAGACAAATACTTCCATGCTCGGGGGAAC TATGATGCTGCCAAAAGGGGACCTGGGGGTGCCTGGGCTGCAGAAGTGAT CACGTAACTGGAGCTCCTGGGACGTTAGGGCTGGGTGAGCAGAGCTTGCC TGCCTTGGACAGTCAGGAGGGAGACGAGCTCCTTGTGGAGAAGTTAGAGG CTGCGGCCCCTCCTCCTCTTGCCCTCTCTCTGCCTCTGTGCTCAGTGTGAGG TCTGAGTGGATGGTAGGAGTGAGTGATTCCTCATCCTCCCTCTCTGGGTGC TGTTCATCCAGCCTAGGGGTGCCCAGCCTGGCTGAATGGGGTGGTGCCCA GTGTTTTCATCCCTCCTTCCTTGGCCTTTCTGGGCTCCTCTCTGAGCCCTCC CTTGGAACAGGGAGAATGGGAGGGTGGGCTATTGCTCACTGGCCTGATTA TTAATCTCCTTCTTGCCTGCCTTGATTACAGCGATGCCAGAGAGAATATCC AGAGATTCTTTGGCCATGGTGCGGAGGACTCGCTGGCTGATCAGGCTGCC AATGAATGGGGCAGGAGTGGCAAAGACCCCAATCACTTCCGACCTGCTGG CCTGCCTGAGAAATACTGAGCTTCCTCTTCACTCTGCTCTCAGGAGATCTG GCTGTGAGGCCCTCAGGGCAGGGATACAAAGCGGGGAGAGGGTACACAA TGGGTATCTAATAAATACTTAAGAGGTGGAATTTGTGGAAA SEQ ID NO:2

[0554] > ENST00000356524.9 SAA1-201 cdna:protein_coding AGGGACCCGCAGCTCAGCTACAGCACAGATCAGCACCATGAAGCTTCTCA CGGGCCTGGTTTTCTGCTCCTTGGTCCTGGGTGTCAGCAGCCGAAGCTTCT TTTCGTTCCTTGGCGAGGCTTTTGATGGGGCTCGGGACATGTGGAGAGCCT ACTCTGACATGAGAGAAGCCAATTACATCGGCTCAGACAAATACTTCCAT GCTCGGGGGAACTATGATGCTGCCAAAAGGGGACCTGGGGGTGCCTGGG CTGCAGAAGTGATCACCGATGCCAGAGAGAATATCCAGAGATTCTTTGGC CATGGTGCGGAGGACTCGCTGGCTGATCAGGCTGCCAATGAATGGGGCAG GAGTGGCAAAGACCCCAATCACTTCCGACCTGCTGGCCTGCCTGAGAAAT ACTGAGCTTCCTCTTCACTCTGCTCTCAGGAGATCTGGCTGTGAGGCCCTC AGGGCAGGGATACAAAGCGGGGAGAGGGTACACAATGGGTATCTAATAA ATACTTAAGAGGTGGAA SEQ ID NO:3

[0555] > SAA1-2O1 cds:protein_coding ATGAAGCTTCTCACGGGCCTGGTTTTCTGCTCCTTGGTCCTGGGTGTCAGC AGCCGAAGCTTCTTTTCGTTCCTTGGCGAGGCTTTTGATGGGGCTCGGGAC ATGTGGAGAGCCTACTCTGACATGAGAGAAGCCAATTACATCGGCTCAGA CAAATACTTCCATGCTCGGGGGAACTATGATGCTGCCAAAAGGGGACCTG GGGGTGCCTGGGCTGCAGAAGTGATCACCGATGCCAGAGAGAATATCCA GAGATTCTTTGGCCATGGTGCGGAGGACTCGCTGGCTGATCAGGCTGCCA ATGAATGGGGCAGGAGTGGCAAAGACCCCAATCACTTCCGACCTGCTGGC CTGCCTGAGAAATACTGA SEQ ID NO: 14

[0556] > SAA1-2O1 peptide: ENSP00000348918 pep:protein_coding PATENT 7328*158807PCT / NLM-1005-PC MKLLTGLVFCSLVLGVSSRSFFSFLGEAFDGARDMWRAYSDMREANYIGSD KYFHARGNYDAAKRGPGGAWAAEVITDARENIQRFFGHGAEDSLADQAANE WGRSGKDPNHFRPAGLPEKY EXAMPLES

[0557] The examples set forth below illustrates certain embodiments and do not limit the technology.

[0558] Example 1: In vitro assessment of test oligomeric compounds

[0559] Oligomeric compounds containing modified oligonucleotides complementary to a human SAA1 RNA, each referred to as an antisense oligonucleotide (ASO), were designed and tested for SAA1 RNA expression inhibition in vitro. The modified oligonucleotides were tested in a series of experiments that had the same culture conditions.

[0560] Each of the modified oligonucleotides of the oligomeric compounds assessed and described in Table 1 and Table 2 below is a 5-10-5 MOE gapmer with mixed phosphorothioate and phosphodiester (PS / PO) internucleoside linkages. Each modified oligonucleotide described in Table 1 and Table 2 (i) is 20 nucleosides in length; (ii) has the following sugar motif (from 5' to 3'): eeeeeddddddddddeeeee, where "e" represents a 2' -MOE ribosyl sugar moiety, and each "d" represents a 2’-deoxyribosyl sugar moiety; (iii) has the following nucleoside linkage motif (from 5' to 3 '): sssssssssssssssssss, where each "s" represents a (Sp)-phosphorothioate internucleoside linkage; and (iv) each “C” nucleoside is a 5-methylcytosine.

[0561] Table 1 defines SAA1 target regions to which oligomeric compounds were complementary. The first column of Table 1 provides a designation for each oligomeric compound. The second and third columns of Table 1 provide the chromosome 11 start position and end position, respectively, for the SAA1 target nucleic acid nucleobase portion to which the corresponding oligomeric compound oligonucleotide is complementary (SAA1 target portion). The fourth and fifth columns of Table 1 provide the start position and end position, respectively, in SEQ ID NO:1 corresponding to chromosome 11 start position and end position provided in columns two and three, respectively, for the SAA1 target portion. The sixth column of Table 1 designates the intron or exon containing the SAA1 target portion (defined by the second, third, fourth and fifth columns). The seventh and eighth columns of Table 1 provide the start position and end position, respectively, in SEQ ID NO:2 corresponding to chromosome 11 start position and end position provided in columns PATENT 7328*158807PCT / NLM-1005-PC two and three, respectively, and corresponding to SEQ ID NO: 1 start and end position provided in columns four and five, respectively, for the SAA1 target portion, as applicable.

[0562] Table 1: SAA1 target portions for oligomeric compounds

[0563]

[0564] +oligomeric compound designation

[0565] * chromosome 11 start position and end position for SAA1 nucleobase portion to which oligomeric compound oligonucleotide is complementary (SAA1 target portion)

[0566] ** start position and end position in SEQ ID NO: 1 corresponding to chromosome 11 start position and end position for SAA1 target sequence in columns 2 and 3 *** start position and end position in SEQ ID NO:2 corresponding to chromosome 11 start position and end position for SAA1 target sequence in columns 2 and 3, as applicable

[0567] Table 2 provides oligonucleotide nucleobase sequences for corresponding oligomeric compounds in Table 1, and in vitro assay screening results. The first column of Table 2 provides a designation for each oligomeric compound. The second column of Table 2 provides the oligonucleotide nucleobase sequence for the corresponding oligomeric compound. Each oligonucleotide nucleobase sequence is 100% complementary to the SAA1 nucleic acid target portion nucleobase sequence defined in Table 1 (in SEQ ID NO: 1). The third column of Table 2 provides a sequence identification number (SEQ ID NO) for the corresponding oligonucleotide nucleobase sequence in the second column. PATENT 7328*158807PCT / NLM-1005-PC Table 2: Oligomeric compound nucleobase sequences and inhibition

[0568]

[0569] oligomeric compound designation

[0570] † oligomeric compound oligonucleotide nucleobase sequence

[0571] sequence identification number (SEQ ID NO) corresponding to oligonucleotide nucleobase sequence in the second column

[0572] ++micromolar IC50value determined for oligomeric compound according to in vitro assay described in this Example

[0573] The fourth column of Table 2 provides the half maximal inhibitory concentration (IC50; micromolar) determined for the corresponding oligomeric compound according to the following in vitro SAA1 assay. Total RNA was prepared using a RNeasy mini kit (Qiagen) from cells grown in 96-well plates based on the manufacturer's protocol. qRT-PCR reactions were performed in triplicates using TaqMan primer probe sets. Approximately 50 nanograms (ng) total RNA in 5 microliters (pL) water was mixed with 0.3 pL primer probe sets containing forward and reverse primers (10 micromolar (pM) of each) and fluorescently labeled probe (3 pM), 0.5 pL reverse transcriptase (RT) enzyme mix (Qiagen), 4.2 pL RNase-free water, and 10 pL of 2* PCR reaction buffer in a 20 pL reaction. Reverse transcription was performed at 48°C for 10 minutes (min), followed by 94°C for 10 min, and then 40 cycles of PCR were conducted at 94°C for 30 seconds (s), and 60°C for 30s within each cycle using the StepOne Plus RT-PCR system (Applied Biosystems). The mRNA levels were normalized using the housekeeping gene GAPDH to the amount of total RNA present in each reaction as determined for duplicate RNA samples. These values were used to calculate the IC50for total SAA1 mRNA, which was quantified using quantitative RT-PCR. PATENT 7328*158807PCT / NLM-1005-PC Example 2: Preparation of oligomeric compound conjugate

[0574] The 5 ’-end of the ASO1 oligomeric compound assessed in Example 1 (see Table 1 and Table 2) was conjugated to a targeting moiety via an intermediary linker. The targeting moiety portion includes a triantenarary N-acetyl-galactosamine (GalNAc) moiety, and is expected to bind to asialoglycoprotein receptor (ASGPR) and enhance ASO1 activity in hepatocytes, the main source of SAA1 protein (Prakash et al., Nucleic Acids Res 42(13):8796-807 (2014)). The ASO1 conjugate prepared and studied is referred to as the “GalNAc-ASOl compound” and “GalNAc-ASOl study drug.”

[0575] The GalNAc-ASOl study drug is an isolated icosasodium salt compound having a formula of C296H424N72Na2oOi5iP2oSi9 and a formula weight of 9095.41 g / mol. The oligonucleotide structure includes five nucleosides at the 5' end and five nucleosides at the 3' end that are modified at the 2'-0 of ribose with a methoxy ethyl (MOE) group. The ten nucleosides in the middle all contain 2'-deoxy ribose. This general structure is referred to as a 5-10-5 MOE gapmer. The GalNAc-ASOl study drug sequence can be written in shorthand as follows:

[0576] 5 '-GalNAc -p-A-mC-mC-mC-A-T-T-G-T-G-T-A-mC-mC-mC-mU-mC-mU-mC-mC-3' (SEQ ID NO : 15)

[0577] All cytosine bases are modified at the 5 position with a methyl group, as are the uracil bases in the MOE nucleosides. The linkage from GalNAc to the oligonucleotide is a phosphodiester, but all internucleoside linkages are phosphorothioates. The underlined nucleosides contain the MOE modification.

[0578] The chemical name of the GalNAc-ASOl study drug can be written as: all-P-amZ>o-5'-O-{[(6-{5-[(tris{3-[6-(2-acetamido-2-deoxy-P-D-galactopyranosyloxy)hexylamino]-3-oxopropoxymethyl})methyl]amino-5-oxopentanamido}hexyl)]phospho}-2'-O-(2-methoxyethyl)-P-thioadenylyl-(3'-O— >5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3'-O—>5'-O)-2'-O-(2-methoxyethyl)-5-methyl- -thiocytidylyl-(3'-O— >5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3 '-O^ 5 '-O)-2'-O-(2-methoxyethyl)-P-thioadenylyl-(3 O—>5 '-O)- -thiothymidylyl-(3 '-O—>5 '-O)- -thiothymidylyl-(3 '-O—>5 '-( )-2'-deoxy- / J-thioguanylyl-(3'-O^5'-O)- -thiothymidylyl-(3 '-( ^5'-( )-2'-deoxy- / J-thioguanylyl-(3 '-O—>5 '-O)-P-thiothymidylyl-(3 '-O—>5 '-O)-2'-deoxy-P-thioadenylyl-(3 '-O—>5 '-O)~ 2'-deoxy-5-methyl-P-thiocytidylyl-(3'-O— >5'-O)-2'-deoxy-5-methyl-P-thiocytidylyl- PATENT 7328*158807PCT / NLM-1005-PC (3'-O^5'-O)-2'-deoxy-5-methyl- -thiocytidylyl-(3'-<9—>5'-<9)-2'-(9-(2-methoxyethyl)-5-methyl-P-thiouridylyl-(3'-(9— >5'-(9)-2'-(9-(2-methoxyethyl)-5- rnethyl- / J-thiocytidylyl-(3'-(9^5'-(9)-2'-<9-(2-methoxyethyl)-5-methyl- / J-thiouridylyl- (3'-O^5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3'-<9— >5'-(9)-2'-(9-(2-methoxyethyl)-5-methylcytidine (SEQ ID NO:20), icosasodium salt. The structure of the GalNac-ASOl study drug is according to the general formula:

[0579]

[0580] In the GalNac-ASOl study drug, the absolute configuration of each 2-<9-(2-methoxyethyl)-D-ribose unit and each 2-(9-(2-deoxy)-D-ribose is (1A, 2A, 3 A, 4R). The absolute configuration of each galactosamine unit is (1A, 2R, 3R, 4R, 5R). The absolute configuration at each phosphorus atom of the phosphorothioate linkages is undefined, and the GalNAc-ASOl study drug is a mixture of 219diastereoisomers. The GalNAc-ASOl study drug was prepared by a process summarized in Figure 1. Manufacture of the GalNAc-ASOl study drug was a multi-step process divided into two distinct operations: solid-phase synthesis and downstream processing. In the first operation, the nucleoside sequence of the GalNAc-ASOl study drug was chemically synthesized in a stainless-steel stationary column which was fed pre-programmed volumes of synthesis reagents by a computer-controlled solid-phase synthesizer. Subsequent downstream processing included deprotection steps, preparative anion exchange (AEX) chromatographic purification, diafiltration, and lyophilization to yield the GalNAc-ASOl study drug. PATENT 7328*158807PCT / NLM-1005-PC During the chemical synthesis, phosphoramidite monomers were sequentially coupled to an elongating oligonucleotide that was covalently bound to a solid support. Each elongation cycle consists of the following four steps:

[0581] 1. Detritylation (removal of a 5 ’-dimethoxytrityl (DMT) hydroxyl protecting group with acid)

[0582] 2. Coupling (attachment of an activated phosphoramidite to the support-bound oligonucleotide)

[0583] 3. Sulfurization (conversion of the newly formed phosphite triester to its phosphorothioate triester with a sulfurizing reagent). All internucleoside linkages are phosphorothioate. The linkage between the UnyLinker solid support and the 3 '-nucleoside is oxidized to the phosphate triester with I2 in pyridine and water, as is the linkage between the 5'-nucleoside and GalNAc.

[0584] 4. Capping (acetylation of unreacted 5 ’-hydroxyls with acetic anhydride).

[0585] This iterative synthesis was executed using the appropriate sequence of nucleoside phosphoramidites until the oligonucleotide sequence was complete. Coupling of the GalNAc targeting moiety was accomplished via the same phosphoramidite process. After the final synthesis step, the oligonucleotide still bound to the solid support was treated with a solution of diethylamine (DEA) in acetonitrile (ACN) to remove the beta-cyanoethyl protecting groups from the phosphorothioate triester linkages. The solid support was washed with diethyl ether to remove residual acetonitrile in preparation for deprotection. The oligonucleotide, still bound to the solid support, was then removed from the synthesis column and treated with ammonium hydroxide in ethanol at elevated temperature. This step simultaneously released the oligonucleotide from the solid support and removed the protecting groups from the bases. The support was removed by filtration and the resulting solution was tested for identity and purity.

[0586] The crude product was then purified by AEX-HPLC. The product was loaded on to the column and eluted from the column with a gradient of 1 M sodium perchlorate (NaClO4) in 0.25 mM Tris base solution (5% MeOH / water). The elution profile was monitored by continuous UV absorption spectroscopy. The product peak was fractionated and the product-containing fractions from each run are tested for purity. Fractions were pooled based on overall purity and yield. The purified product included the selected fractions and then was desalted by diafiltration. The final product was filtered through a 0.2-micron filter and lyophilized. The material was PATENT 7328*158807PCT / NLM-1005-PC transferred into HDPE bottles with polypropylene lids that have been gammairradiated. The bottles were sealed and labeled.

[0587] Example 3: Safety and tolerability assessments of SSA1 oligomeric compound conjugate

[0588] The GalNAc-ASOl study drug was evaluated for safety and tolerability. The fully protonated form of the GalNAc-ASOl compound described in Example 2 has a formula of C296H444N72O151P20S19. The formula weight of the fully protonated form of the GalNAc-ASOl compound is 8655.78 g / mol. This is the theoretical molecular weight detected by mass spectrometry. The fully protonated form of the parent ASO1 oligonucleotide without the GalNAc targeting moiety and linker has a formula of C230H325N64O122P19S19 and the formula weight is 7136.10 g / mol. In conformance with USP and FDA guidance, pre-clinical dosing (in this Example 3) and clinical dosing (in Example 4) was based on the concentration of the fully protonated parent ASO1 oligonucleotide, which for clarity is according to the following general formula:

[0589]

[0590] PATENT 7328*158807PCT / NLM-1005-PC Oligomeric compound safety and tolerability assessment in C57B / 6 Mice Mice were dosed with 50 mg / kg of GalNAc-ASOl two times per week for 4 weeks (n=3 per oligonucleotide tested). Animals were monitored for body weight and clinical signs during the treatment period. Two days after the last dose, plasma was collected from the mice and analyzed for changes in serum chemistry parameters. Mice were euthanized and liver, kidney and spleen tissue collected and analyzed for histological changes.

[0591] There were no clinical observations in the mice treated with GalNAc-ASOl and all mice exhibited normal body weight increases. Liver, kidney, and spleen weights were within 10% of saline treated control mice. Serum chemistry parameters, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin, albumin, blood urea nitrogen and creatine were not significantly different than saline treated animals. Based on a 4-week repeat dose high dose study in C57B / 6 mice, GalNAc-ASOl was well tolerated at a dose of 50 mg / kg administered twice weekly with no safety concerns.

[0592] Oligomeric compound assessment in human SAA1 transgenic mice

[0593] Human SAA1 transgenic mice were generated by transgenesis of a fosmid containing the entire human SAA1 gene including its promoter. The basal levels of human SAA1 mRNA in liver and serum protein was similar to basal levels of mouse SAA1 mRNA and protein. Both human and mouse SAA1 expression was increased by treatment of the transgenic mice with azocasein (inflammatory agent used to induce amyloid A amyloidosis), suggesting that it is a response to inflammatory stimuli. Activity of GalNAc-ASOl was evaluated in mice stimulated with azocasein. The treatment of human SAA1 transgenic mice with 3 doses (every 3 days) of GalNAc-ASOl produced a dose dependent reduction of human SAA1 mRNA in liver. Mice treated with 0.1 and 1 mg / kg of GalNAc-ASOl exhibited approximately 50% and 90% reduction of human SAA1 mRNA, respectively. Treatment of transgenic mice with GalNAc-ASOl was well tolerated.

[0594] Toxicology Studies

[0595] Potential toxicity of GalNAc-ASOl was assessed in a 5-week dose rangefinding (DRF) study and a GLP compliant 13 -week repeat dose study in CD-I mice with the test item administered at weekly intervals. An additional loading dose was PATENT 7328*158807PCT / NLM-1005-PC provided on Day 4 in the 5-week study. A summary of completed repeat-dose toxicity studies is provided in Table 3 below.

[0596] Table 3: Summary of GalNAc-ASOl Toxicology Studies

[0597]

[0598] Abbreviations: ASO= Antisense Oligonucleotide, GLP= Good Laboratory Practices, SC = Subcutaneous

[0599] A. 5-Week Repeat Subcutaneous Dose Toxicity Study with GalNAc-ASOl and Mouse Surrogate m-ASO in CD-I Mice

[0600] Objective

[0601] The objective of this study was to evaluate the potential toxicity of GalNAc-ASOl and a mouse surrogate oligomeric compound in CD-I mice after 5 weeks of subcutaneous (SC) administration. The data from this study was used to inform the dose selection for subsequent 13-week repeat-dose GLP toxicology study.

[0602] Since GalNAc-ASOl is not pharmacologically active in mice, a mouse-specific inhibitor of Saa referred to as “m-ASO,” was included to evaluate the potential effects associated with exaggerated pharmacology. The mouse surrogate oligomeric compound specifically targets the mouse Saal gene and is a 5-10-5 MOE gapmer with mixed phosphorothioate and phosphodiester (PS / PO) internucleoside linkages. The m-ASO mouse surrogate oligomeric compound (i) is 20 nucleosides in length; (ii) has the nucleobase sequence (from 5’ to 3’): TGTTTATTACCCTCTCCTCC (SEQ ID NO: 13); (iii) has the following sugar motif (from 5' to 3'): eeeeeddddddddddeeeee, where "e" represents a 2' -MOE ribosyl sugar moiety, and each "d" represents a 2’-deoxyribosyl sugar moiety; (iv) has the following nucleoside linkage motif (from 5' to 3'): sssssssssssssssssss, where each "s" represents a (S)-phosphorothioate internucleoside linkage; and (iv) each “C” nucleoside is a 5-methylcytosine. PATENT 7328*158807PCT / NLM-1005-PC Method

[0603] Six male animals / group were assigned to treatment groups receiving weekly SC doses of PBS or GalNAc-ASOl at 50 and 80 mg / kg / week. In addition, four male animals / group received m-ASO at 1, 5, and 10 mg / kg / week (with a loading dose on Day 4). Terminal sacrifice was performed on Day 40, approximately 96 hours after final dosing on Day 36.

[0604] Table 4: Study Design

[0605]

[0606] aAdministered once weekly with one loading dose on Day 4.

[0607] bControl and test items were administered via subcutaneous injection with necropsies occurring on Day 40 (terminal necropsy, 96 hours after final dose on Day 36).

[0608] cMouse specific oligonucleotide.

[0609] Results

[0610] In the 5-week non-GLP study, mice were treated with 50 and 80 mg / kg / week GalNAc-ASOl for 5 weeks, with one additional loading on Day 4 in the first week. The GalNAc-ASOl oligomeric compound was well tolerated in CD-I mice when administered subcutaneously for 5 weeks at doses up to 80 mg / kg / week, with no test item-related effects on mortality, clinical observations, body weight, or organ weights. GalNAc-ASOl -related effects were limited to alterations in hematology, clinical chemistry, and histopathology. Minimal elevations in absolute neutrophils and eosinophils were noted in mice receiving 80 mg / kg / week GalNAc-ASOl. Minimal increases in absolute monocytes were observed in animals treated with > 50 mg / kg / week GalNAc-ASOl. Minimal elevations in ALT (up to 2.3-fold over control) and AST (up to 1.8- fold over control) were noted in the mice treated with greater PATENT 7328*158807PCT / NLM-1005-PC than or equal to 50 mg / kg / week GalNAc-ASOl. These minimal elevations in ALT and AST correlated with microscopic findings in hepatocyte and Kupffer cells, including hepatocyte karyomegaly, increased hepatocyte mitotic activity, basophilic granules within Kupffer cells, hepatocyte hypertrophy, and single cell hepatocyte necrosis (80 mg / kg / week GalNAc-ASOl only). These findings were minimal in severity and attributed to oligomeric compound uptake and retention. In addition to the aforementioned findings in hepatocytes and Kupffer cells, mild mixed inflammatory cell infiltrates in the liver were noted in mice that had received 80 mg / kg / week of GalNAc-ASOl. These results suggested an ASO-related pro-inflammatory response and correlated with the minimal increases in absolute neutrophils, monocytes, and eosinophils identified in hematology evaluation.

[0611] Table 5: Summary of Clinical Pathology Findings

[0612]

[0613] No test item-related effects in clinical pathology, organ weight, or microscopic findings were observed with m-ASO, at doses up to 10 mg / kg / week for 5 weeks. Up to 82% inhibition in hepatic Saal mRNA expression was observed at 10 mg / kg / week m-ASO after 5 weeks of treatment. There were no specific findings that were associated with this level of pharmacologic inhibition of hepatic Saal expression. PATENT 7328*158807PCT / NLM-1005-PC In conclusion, the toxicities observed with GalNAc-ASOl were generally dose-related and were largely due to oligonucleotide tissue accumulation and the known proinflammatory effects of oligonucleotide treatment in rodents. Those findings were considered non-adverse due to their low incidence, magnitude / severity, and / or lack of functional effects.

[0614] B. 13-week GLP repeat subcutaneous dose toxicity study with GalNAc-ASOl in CD-I Mice

[0615] Objective

[0616] The objective of this study was to evaluate the potential toxicity and tissue concentration of GalNAc-ASOl in CD-I mice after 13 weeks of subcutaneous (SC) administration. Since GalNAc-ASOl is not pharmacologically active in mice, the mouse-specific inhibitor of Saa m-ASO, was included to evaluate the potential effects associated with exaggerated pharmacology.

[0617] Method

[0618] Ten mice / sex / group were assigned to treatment groups receiving weekly SC doses of saline, GalNAc-ASOl at 10, 30 and 60 mg / kg / week, and m-ASO, the mouse-specific Saal ASO, at 10 mg / kg / week. Terminal sacrifice was performed on Day 94, approximately 48 hours after final dosing on Day 92.

[0619] Table 6: Study Design PATENT 7328*158807PCT / NLM-1005-PC

[0620]

[0621] aControl and test items were administered once weekly via subcutaneous injection, with necropsies occurring on Day 92 (terminal necropsy, 48 hours after final dose on Day 92).

[0622] bMouse specific oligonucleotide.

[0623] Results

[0624] There were no test item-related changes based on mortality, clinical observations, body weight, food consumption and macroscopic findings. GalNAc-ASO1 -related microscopic findings were identified in the liver, injection site, skin / inguinal, and multiple tissues.

[0625] Microscopic changes within the liver were identified in both sexes and were characterized by minimal to slight single cell necrosis at dose levels greater than or equal to 30 mg / kg / week and basophilic granules within Kupffer cells at 60 mg / kg / week. The single cell necrosis within the liver correlated with minimal changes in serum biochemical liver parameters (Table 7), including increases in total bilirubin (TBIL) in males at greater than or equal to 30 mg / kg / week (up to 1.5-fold) and females at 60 mg / kg / week (1.1-fold), increases in aspartate aminotransferase (AST) in males at 60 mg / kg / week (up to 2.1-fold) and females at greater than or equal to 30 mg / kg / week (1.2-fold), and elevated alanine aminotransferase (ALT) in males at 60 mg / kg / week (1.4-fold). A minimal increase in gamma-glutamyl transferase (GGT) was also observed in males at 60 mg / kg / week. It should be noted that one male mouse in the 60 mg / kg / week group demonstrated values of AST and ALT that were higher than the rest of the mice within the group. The creatine kinase value for this mouse was also elevated, suggesting possible procedure-related peri-mortem muscle injury. PATENT 7328*158807PCT / NLM-1005-PC Therefore, the muscle isozyme may have contributed to the elevated AST and ALT in this animal.

[0626] Microscopic changes within the injection site, skin / inguinal, and various other tissues (including, but not limited to the heart, pancreas, lymph nodes [mandibular, mesenteric, inguinal, and iliac], seminal vesicle [with coagulation gland], prostate, epididymis, testes, uterus / cervix, and synovium of the fem orotibial joint) were characterized by the presence of vacuolated / granular macrophages. The vacuolated / granular macrophages were consistent with lysosomal uptake, accumulation, and retention of the oligonucleotide test item.

[0627] Minimal mononuclear cell infiltration was identified at the injection site of both sexes at greater than or equal to 10 mg / kg / week. This change was likely related to a minimal localized oligonucleotide specific inflammatory response.

[0628] In addition to the aforementioned minimal changes in liver parameters, in GalNAc-ASO1 -treated mice, clinical chemistry evaluation also indicated minimal decreases in total protein (TP) and albumin (ALB) with no concurrent changes in globulin (GLO) in males at greater than or equal to 10 mg / kg / week and females at 60 mg / kg / week (Table 7). Such a change is suggestive of an acute phase inflammatory response. On a similar note, hematology evaluation showed mild increases in absolute white blood cells (WBC), neutrophils (NEU), lymphocytes (LYMA), monocytes (MONA), eosinophils (EOS), and large unstained cells (LUC) in females at greater than or equal to 10 mg / kg / week and males at either greater than or equal to 10 mg / kg / week (NEU and MONA) or greater than or equal to 30 mg / kg / week (WBC, LYMA, and LUC) which were consistent with a minimal acute inflammatory response (such as an ASO associated inflammatory response, although no histological correlate was identified). PATENT 7328*158807PCT / NLM-1005-PC Table 7: Clinical chemistry results

[0629]

[0630] M=Male and F=Female

[0631] *R = Dunn Rank Sum Test Significant at the 0.05 level

[0632] +D = Dunnett LSD Test Significant at the 0.01 level

[0633] *D = Dunnett LSD Test Significant at the 0.05 level

[0634] +T = Student T Test Significant at the 0.01 level

[0635] *T = Student T Test Significant at the 0.05 level

[0636] +r = Wilcoxon Rank Sum Test Significant at 0.01 level

[0637] aThis is mean value. Since the control value is 0, it cannot be used as a fold.

[0638] Treatment with GalNAc-ASOl and the m-ASO at 10 mg / kg / week for 13 weeks produced about 89% and 94% reduction in hepatic SAA1 mRNAfrom males and females, respectively. The findings with m-ASO treatment were mostly comparable to those observed with GalNAc-ASOl. There were no specific findings that were associated with this level of pharmacologic inhibition of hepatic Saal mRNA expression.

[0639] A dose-dependent increase in liver and kidney exposure was confirmed in male and female mice over the dose range of 10 to 60 mg / kg / week following 13 weeks of treatment. The mean tissue concentrations of unconjugated ASO1 and GalNAc-ASOl in the liver and kidney after administration at 60 mg / kg / week were PATENT 7328*158807PCT / NLM-1005-PC 510.8 ± 47.6 pg / g and 160.2 ±69.3 pg / g for males, respectively and 523.8 ± 103.9 pg / g and 520.5 ± 379.8 pg / g for females, respectively.

[0640] The concentration of unconjugated ASO1 in liver and kidney increased in a dose dependent manner, with a nearly dose-proportional increase observed in liver (approximately 5.7-fold and 4.9-fold in males and females, respectively), but a less than dose-proportional increase observed in kidney (approximately 3.1-fold and 2.9-fold in males and females, respectively) over the 6-fold dose range evaluated from 10 to 60 mg / kg / week. Overall, there were no notable sex differences in the liver exposure; however, there was a generally dissimilar trend in kidney exposure, where females showed slightly higher levels relative to males at all dose groups (up to approximately 3.5-fold). At terminal sacrifice (approximately 48 hours post the last dosing on Day 92), the concentration of GalNAc-ASOl in the liver and kidney was below the lower limit of quantitation.

[0641] In conclusion, subcutaneous administration of GalNAc-ASOl at 10, 30 and 60 mg / kg / week to CD-I mice for 13 weeks resulted in changes in clinical chemistry, hematology, spleen and mandibular lymph nodes weights and correlated with microscopic findings. These changes were considered non-adverse since there were no correlating findings in clinical observations, body weights, or food consumption, and since the magnitudes of changes were minimal to slight and were attributed to the oligomeric compound-related tissue accumulation and / or pro-inflammatory effects. The no-observed-adverse effect level (NOAEL) for GalNAc-ASOl in mice was determined to be 60 mg / kg / week based on the 13 -week repeat-dose study.

[0642] Example 4: Clinical assessment of SSA1 oligomeric compound conjugate

[0643] A patient treated with GalNAc-ASOl (study drug) was a 42-year-old male who has biopsy-proven serum amyloid A (SAA) amyloidosis in the kidney. The patient’s sister and mother also had been diagnosed with AA amyloidosis. Genetic testing for familial periodic fever syndromes were negative; however, a variant chrl 1: 18290751 A> T (hg 19) encoding for a missense mutation in the SAA1 gene (NM_199161.5):c.l01A> T(p. D34V) was identified in both siblings (N. Leung et al., Kidney International, In process (2024)). The mutation is thought to accelerate the misfolding, precipitation, and aggregation of SAA1. The patient’s mother died from complications of AA amyloidosis, and his maternal grandfather died of kidney failure. The patient’s sister presented at 31 years of age with a similar profile. Despite PATENT 7328*158807PCT / NLM-1005-PC immunosuppressive treatment, the sister’s renal function deteriorated, and she developed a spontaneous left renal hematoma followed by a right spontaneous hematoma, subsequently suffered hemorrhagic shock necessitating bilateral nephrectomy, and ultimately died from the disease.

[0644] Study Design

[0645] Before starting the study, the patient had chronic kidney disease stage 3 A with an estimated glomerular filtration rate (eGFR) of 52 ml / min / 1.73 m2. The patient’s eGFR reduced by 5-6 ml / min / 1,73m2per year for the two years prior to treatment start. The patient’s proteinuria was averaging 3500 mg / d but recently had decreased prior to the treatment start. Renal biopsy demonstrated amyloid deposits. SAA1 levels were normal, and no elevation of pro-inflammatory mediators was found.

[0646] Following informed consent, and after baseline clinical laboratory and data collection, the study drug was administered subcutaneously, using a monthly dosing schedule. The study was planned for 12 months. The primary treatment goals included reduction of the rate of deterioration of renal function as measured with estimated glomerular filtration rate (eGFR), as well as prevention to initiate dialysis or require kidney transplantation. The secondary treatment goal included quality of life improvement assessed by a quality-of-life questionnaire. An exploratory treatment goal was to reduce levels of plasma SAA1 protein as measured by specific SAA1 assay. Serum concentrations of SAA were quantified using the latex-enhanced immunonephelometry Siemens N Latex SAA Kit (Siemens Healthcare Diagnostics Products, Germany) and Human SAA ELISA Kit (ThermoFisher # KHA0011). The samples were run in at least 3 replicate assays. The study set assessments were specified to be completed at every visit and analyzed at the 12-month timepoint. The study set safety and tolerability assessments were specified to be completed at every visit throughout the study. The Results presented hereafter are at the 18-month timepoint. The patient has completed the 12-month study and has entered the maintenance phase.

[0647] Results

[0648] Treatment was initiated at 40 mg of the study drug administered subcutaneously. After 2 doses at 40 mg, the dose was escalated to 80 mg for 3 doses, and then to 100 mg. The patient received 13 doses at 100 mg, for a total of 18 doses to date. PATENT 7328*158807PCT / NLM-1005-PC The study drug was well tolerated, and the patient did not experience an ASO-related adverse event or ASO-related serious adverse event. A rapid reduction in plasma SAA1 levels, a reduction in proteinuria, and stabilization in his eGFR values were observed in response to the study drug for the patient.

[0649] As shown in Table 8 (see FIG. 3), the eGFR levels at baseline were 55 mL / min / 1.73m2, which is considered “Stage 3 (30-59): Mild to moderate loss of kidney function”. Since treatment start, the eGFR values have stabilized (not showing a further decrease), and even increased to 68 mL / min / 1.73m2, which is considered “Stage 2 (60-89): Kidney damage with mild loss of kidney function.”

[0650] Figure 2 shows the sharp reduction in plasma SAA1 levels, to levels close to the upper limit of assay sensitivity, emphasizing target engagement. Samples are batched, and Month 13 to Month 18 samples will be analyzed at a later timepoint. Overall, based on the clinical labs and biomarker assessments, this patient’s disease is showing trends towards stabilization, supporting the patient’s benefit / risk ratio. Non-limiting disclosure and incorporation by reference

[0651] The entirety of each patent, patent application, publication, document, sequence, sequence information (according to database accession number or identifier (for example, GenBank, ENSEMBL and dbSNP database identifiers) for example) and the like cited herein (collectively “citations”), is incorporated by reference. A citation is not an admission that any of the foregoing is pertinent prior art, nor does it constitute any admission as to the contents or date of publications or documents. Their citation is not an indication of a search for relevant disclosures. All statements regarding the date(s) or contents of the documents is based on available information and is not an admission as to their accuracy or correctness. Incorporation by reference of documents, standing alone, should not be construed as an assertion or admission that any portion of the contents of any document is considered to be essential material for satisfying any national or regional statutory disclosure requirement for patent applications. Notwithstanding, the right is reserved for relying upon any of such documents, where appropriate, for providing material deemed essential to the claimed subject matter by an examining authority or court.

[0652] The technology is described with reference to specific embodiments and nonlimiting Examples. The terms and expressions utilized herein to describe the technology are descriptive and not necessarily limiting. Certain modifications made to PATENT 7328*158807PCT / NLM-1005-PC the disclosed embodiments can be considered within the scope of the technology. Certain aspects of the disclosed embodiments suitably can be practiced in the presence or absence of certain elements not specifically disclosed herein.

[0653] Certain nucleobase sequences provided can be within a nucleic acid having any combination of chemical modifications described herein. For example, although a sequence listing identifies each sequence as either “RNA” or “DNA” as required, in reality, a nucleic acid comprising a referenced nucleobase sequence can include any combination of chemical modifications described herein. The designations “RNA” or “DNA” is arbitrary in certain instances. For example, an oligonucleotide comprising a nucleoside comprising a 2 -OH sugar moiety and a thymine base can be described as a DNA having a modified sugar (2 ’-OH in place of one 2’-H of DNA) or as an RNA having a modified base (thymine (methylated uracil ) in place of an uracil of RNA). Also, for example, an oligomeric compound having the nucleobase sequence “ATCGATCG” encompasses a modified or unmodified oligomeric compound having the nucleobase sequence, including compounds comprising RNA bases (having the sequence “AUCGAUCG” for example), compounds having some DNA bases and some RNA bases (having the nucleobase sequence “AUCGATCG” for example) and compounds having one or more modified nucleobases (having the nucleobase sequence “AT’"CGAUCG,” wherefnC indicates a cytosine base comprising a methyl group at the 5-position). Accordingly, nucleobase sequences provided, including those in a sequence listing, are intended to encompass nucleic acids containing any combination of natural or modified RNA and / or DNA, including nucleic acids having m modified nucleobases.

[0654] Certain compounds described herein (for example, modified oligonucleotides) have one or more asymmetric center and thus give rise to enantiomers, diastereomers, and other stereoisomeric configurations that can be defined, in terms of absolute stereochemistry, as (R) or (S), as alpha or beta such as for sugar anomers, or as (D) or (L), such as for amino acids, and the like. Compounds provided that are drawn or described as having certain stereoisomeric configurations include only the indicated compounds. Compounds provided that are drawn or described with undefined stereochemistry include all such possible isomers, including their stereo-random and optically pure forms, unless specified otherwise. Tautomeric forms of the compounds PATENT 7328*158807PCT / NLM-1005-PC herein are also included unless otherwise indicated. Unless otherwise indicated, compounds described herein are intended to include corresponding salt forms.

[0655] Certain compounds described herein include variations in which one or more atoms are replaced with a nonradioactive isotope or radioactive isotope of' the indicated element. For example, compounds herein that comprise hydrogen atoms encompass all possible deuterium substitutions for each of thefH hydrogen atoms. Isotopic substitutions encompassed by the compounds herein include but are not limited to:2H or3H in place of1H,11C or14C in place of12C,15N in place of14N,17O or18O in place

[0656]

[0657] 35S, or36S in place of32S. In certain embodiments, nonradioactive isotopic substitutions may impart new properties on the oligomeric compound that are beneficial for use as a therapeutic or research tool. In certain embodiments, radioactive isotopic substitutions may make the compound suitable for research or diagnostic purposes such as imaging.

[0658] Each of the terms “comprising,” “consisting essentially of,” and “consisting of’ can be replaced with either of the other two terms. The terms “including,” “include,” “includes” and “included” are not limiting and can be replaced with the term “comprising” or grammatic variant thereof. As used in this specification and the claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. The use of the singular herein includes the plural unless specifically stated otherwise. The use of “or” means “and / or” unless stated otherwise, and unless specifically stated or obvious from context, as used herein, the term “or” is understood to be inclusive and covers both “or” and “and.” The term “one or more” means “at least one” and the term “at least one” means “one or more.” The terms “element” or “component” encompass elements and components including one unit and elements and components that include more than one subunit, unless specifically stated otherwise. The term “exemplary” means “an example of’ or “examples of’ according to context.

[0659] Unless specifically stated or obvious from context, as used herein, the terms “substantially all”, “substantially most of’, “substantially all of’ or “majority of’ encompass at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5%, or more of a referenced amount of a composition. Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 PATENT 7328*158807PCT / NLM-1005-PC standard deviations of the mean. The term “about” can be understood as within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from the context, all numerical values provided herein are modified by the term “about.” Use of the term “about” at the beginning of a listing of values modifies each of the values (e.g., “about 1, 2 and 3” refers to "about 1, about 2 and about 3"). When a listing of values is described the listing includes all intermediate values and all fractional values thereof (e.g., the listing of values "80%, 85% or 90%" includes the intermediate value 86% and the fractional value 86.4%). When a listing of values is followed by the term "or more," the term "or more" applies to each of the values listed (e.g., the listing of "80%, 90%, 95%, or more" refers to "80% or more, 90% or more, or 95% or more"). When a listing of values is described, the listing includes all ranges between any two of the values listed (e.g., the listing of "80%, 90% or 95%" includes ranges of "80% to 90%, " "80% to 95%" and "90% to 95%").

[0660] A number of embodiments of the invention have been described.

[0661] Nevertheless, it can be understood that various modifications may be made without departing from the spirit and scope of the invention. Any of the above aspects and embodiments can be combined with any other aspect or embodiment as disclosed here in the Summary, Figures and / or Detailed Description sections. Accordingly, other embodiments are within the scope of the following claims.

Claims

1. PATENT 7328*158807PCT / NLM-1005-PC WHAT IS CLAIMED IS:

1. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to an equal length portion of a SAA1 nucleic acid, and wherein the modified oligonucleotide comprises at least one modification selected from a modified sugar moiety and a modified internucleoside linkage.

2. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and comprising or consisting of a nucleobase sequence containing at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases complementary to:4.an equal length portion of nucleobases 3458-3708, 3624-3727 or 3644-3707 of SEQ ID NO: 1;5.an equal length portion of nucleobases 3644-3663, 3647-3666, 3652-3671, 3663-3682, 3666-3685, 3677-3696 or 3688-3707 of SEQ ID NO: 1;6.an equal length portion of nucleobases 3643-3702, 3643-3692 or 3663-3682 of SEQ ID NO: 1;7.an equal length portion of nucleobases 720-3074, 2908-2967 or 2928-2947 of SEQ ID NO: 1; or8.an equal length portion of nucleobases 720-2935, 2200-2259 or 2220-2239 of SEQ ID NO: 1;9.wherein the modified oligonucleotide comprises at least one modification selected from a modified sugar moiety and a modified internucleoside linkage.

3. The oligomeric compound of claim 2, wherein the modified oligonucleotide comprises or consists of a nucleobase sequence that is at least 85%, at least 90%, at least 95%, or 100% complementary to an equal length portion of a SAA1 nucleic acid, wherein the SAA1 nucleic acid has the nucleobase sequence of SEQ ID NO: 1 or SEQ ID NO:2.

4. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and comprising or consisting of a nucleobase sequence containing at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at leastPATENT 7328*158807PCT / NLM-1005-PC 18, at least 19, or 20 contiguous nucleobases of any of (i) SEQ ID NO:4 to SEQ ID NO: 12, (ii) SEQ ID NO:4 to SEQ ID NO:7 and SEQ ID NO:9 to SEQ ID NO: 11, (iii) SEQ ID NO:8, (iv) SEQ ID NO: 12 or (v) SEQ ID NO:4, wherein the modified oligonucleotide comprises at least one modification selected from a modified sugar moiety and a modified internucleoside linkage.

5. The oligomeric compound of any one of claims 1-4, wherein the modified oligonucleotide comprises at least one modified internucleoside linkage, and optionally wherein the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage.

6. The oligomeric compound of claim 5, wherein the modified oligonucleotide comprises an intemucleoside linkage motif (from 5' to 3’) selected from sososssssssssssooss, sooosssssssssssooss, soooossssssssssooss or sssssssssssssssssss, wherein each “s” represents a phosphorothioate intemucleoside linkage and each “o” represents a phosphodiester intemucleoside linkage, and optionally each “s” represents a (Sp)-phosphorothioate intemucleoside linkage.

7. The oligomeric compound of any one of claims 1-6, wherein the modified oligonucleotide comprises at least one modified nucleoside, optionally wherein the modified nucleoside comprises a comprises a modified sugar, and optionally wherein the modified sugar comprises a 2’-M0E sugar moiety.

8. The oligomeric compound of any one of claims 1-7, wherein the modified oligonucleotide comprises:16.a 5’-region consisting of 1-6 linked 5’-region nucleosides;17.a central region consisting of 6-10 linked central region nucleosides; and a 3 ’-region consisting of 1-6 linked 3 ’-region nucleosides; wherein: each of the 5 ’-region nucleosides and each of the 3 ’-region nucleosides comprises a modified sugar moiety, and three or more of the central region nucleosides comprise a 2’-beta-D-deoxyribosyl sugar moiety.

9. The oligomeric compound of any one of claims 1-8, wherein the modified oligonucleotide comprises:19.a 5 ’-region consisting of 5 linked 5 ’-region nucleosides;20.a central region consisting of 10 linked central region nucleosides; and a 3 ’-region consisting of 5 linked 3 ’-region nucleosides; wherein: PATENT 7328*158807PCT / NLM-1005-PC each of the 5 ’-region nucleosides and each of the 3 ’-region nucleosides comprises a modified sugar moiety, and three or more of the central region nucleosides comprise a 2’-beta-deoxyribosyl sugar moiety.

10. The oligomeric compound of any one of claims 1-9, wherein the modified oligonucleotide comprises a sugar motif of 5’-eeeeeddddddddddeeeee-3’, wherein each "d ’’ represents a 2’-beta-D-deoxyribosyl sugar moiety, each “e” represents a 2’-MOE sugar moiety.

11. The oligomeric compound of any one of claims 1-10, wherein: the modified oligonucleotide (i) consists of 20 nucleosides; (ii) consists of a sugar motif (from 5' to 3') of eeeeeddddddddddeeeee, wherein "e" represents a 2' -MOE ribosyl sugar moiety, and each "d" represents a 2’ -deoxyribosyl sugar moiety; (iii) consists of a nucleoside linkage motif (from 5' to 3 ') of sssssssssssssssssss, wherein each "s" represents a (Sp)-phosphorothioate internucleoside linkage; and (iv) each “C” nucleoside is a 5-methylcytosine.

12. The oligomeric compound of any one of claims 1-11, wherein the oligomeric compound comprises a conjugate group.

13. The oligomeric compound of claim 12, wherein the conjugate group comprises a targeting moiety and a linker moiety.

14. The oligomeric compound of claim 13, wherein the targeting moiety comprises a carbohydrate ligand; optionally wherein the carbohydrate ligand comprises or consists of a N-acetylgalactosamine (GalNAc) moiety; optionally wherein the targeting moiety comprises multiple GalNAc moieties, a branching group and a tether moiety linking the branching group to each of the GalNAc moieties; optionally wherein the targeting moiety comprises two, three or four GalNAc moieties, and optionally three GalNAc moieties; optionally wherein the targeting moiety comprises or consists of a structure according to the following general Formula A:PATENT 7328*158807PCT / NLM-1005-PC27. 29.Formula A30.and each m and each n in Formula A independently is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; and optionally wherein each m independently is an integer of 5, 6 or 7 and each n independently is an integer of 1, 2 or 3 in Formula A.

15. The oligomeric compound of claim 13 or claim 14, wherein the linker moiety comprises or consists of a structure according to one of the following general Formula B, Formula C or Formula D:

35. 37.and each n independently is zero or an integer of 1, 2, 3, 4, 5, 6 or 7 in Formula B or Formula C; optionally wherein each n independently is an integer of 1, 2, 3, 4 or 5 in Formula B or Formula C; and optionally wherein each n independently is the integer 2 or 5 in Formula B or Formula C. PATENT 7328*158807PCT / NLM-1005-PC 16. The oligomeric compound of any one of claims 13-15, wherein the targeting moiety and linker moiety together are according to the following general formula:

39.

40.

17. The oligomeric compound of any of claims 1-16, wherein the modified oligonucleotide consists of 12 to 20, 12 to 25, 12 to 30, 13 to 20, 13 to 25, 13 to 30, 14 to 20, 14 to 25, 14 to 30, 15 to 20, 15 to 25, 15 to 30, 16 to 18, 16 to 20, 16 to 25, 16 to 30, 17 to 20, 17 to 25, 17 to 30, 18 to 20, 18 to 25, 18 to 30, 19 to 20, 19 to 25, 19 to 30, 20 to 25, or 20 to 30, linked nucleosides.

18. The oligomeric compound of any one of claims 1-17, wherein the modified oligonucleotide consists of (i) a nucleobase sequence containing at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases complementary to an equal length portion of nucleobases 3643-3702, 3643-3692 or 3663-3682 of SEQ ID NO:1; or (ii) a nucleobase sequence containing at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of SEQ ID NO:4.

19. The oligomeric compound of claim 18, wherein the modified oligonucleotide consists of the nucleobase sequence of SEQ ID NO:4.

20. An oligomeric compound, comprising or consisting of all-P-ambo-5'-O-{[(6-{5-[(tris{3-[6-(2-acetamido-2-deoxy-P-D-galactopyranosyloxy)hexylamino]-3-oxopropoxymethyl})methyl]amino-5-oxopentanamido}hexyl)]phospho}-2'-O-(2-methoxyethyl)-P-thioadenylyl-(3'-O^5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3'-O— >5'-O)-2'-O-(2 -methoxy ethyl)-5-methyl-P-thiocytidylyl -(3'-O^5'-O)-2'-O-(2-methoxyethyl)-5-methyl- -thiocytidylyl-(3 '-0^>-5'-0)-2'-0-(2-methoxyethyl)- -thioadenylyl-(3'-O^5'-O)- -thiothymidylyl-(3'-O— >5'-O)- -PATENT 7328*158807PCT / NLM-1005-PC thiothymidylyl-(3'-O^5'-O)-2'-deoxy- -thioguanylyl-(3'-O—>5'-O)- -thiothymidylyl-(3'-O^5'-O)-2'-deoxy-P-thioguanylyl-(3'-O—>5'-O)-P-thiothymidylyl-(3'-O— >5'-O)-2'-deoxy-P-thioadenylyl-(3'-O— >5'-O)-2'-deoxy-5-methyl-P-thiocytidylyl-(3'-O— >5'-O)-2'-deoxy-5-methyl-P-thiocytidylyl-(3'-O— >5'- O)-2'-deoxy-5-methyl-P-thiocytidylyl-(3'-O— >5 '-O)-2'-O-(2 -methoxy ethyl)-5-methyl-P-thiouridylyl-(3'-O^5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3'- O^5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiouridylyl-(3'-O—>5'-O)-2'-O-(2-methoxyethyl)-5-methyl-P-thiocytidylyl-(3'-O— >5'-O)-2'-O-(2-methoxyethyl)-5-methylcytidine (SEQ ID NO:20), icosasodium salt.

21. An oligomeric compound according to the general formula:

46.

22. A composition comprising an oligomeric compound of any one of claims 1-21, wherein the composition optionally is a pharmaceutical composition comprising an oligomeric compound of any one of claims 1-21 and a pharmaceutically acceptable diluent.

23. An oligomeric compound of any one of claims 1-21, or a composition or pharmaceutical composition of claim 22, for reducing SAA1 expression.PATENT 7328*158807PCT / NLM-1005-PC 24. An oligomeric compound of any one of claims 1-21, or a composition or pharmaceutical composition of claim 22, for treating a medical condition, wherein the medical condition optionally is a SAA1 -associated medical condition, and wherein the SAA1 -associated medical condition optionally is amyloid A amyloidosis, renal failure, autonomic neuropathy, congestive heart failure or arrhythmia.

25. The oligomeric compound, composition or pharmaceutical composition of claim 24, wherein at least one symptom or hallmark of an SAA1 -associated medical condition is ameliorated, wherein the symptom or hallmark optionally is one or more of proteinuria, nephrotic syndrome, low blood protein level, low blood albumin level, edema, high cholesterol, enlarged spleen, enlarged liver, enlarged thyroid, orthostatic hypotension, gastrointestinal atony, diarrhea and constipation.

26. The oligomeric compound, composition or pharmaceutical composition of any one of claims 23-25, wherein an elevated SAA1 level is reduced.

27. The oligomeric compound, composition or pharmaceutical composition of claim 26, wherein the elevated SAA1 level is an elevated level of a SAA1 nucleic acid, or SAA1 protein or SAA1 nucleic acid and SAA1 protein.

28. The oligomeric compound, composition or pharmaceutical composition of claim 26 or claim 27, wherein the elevated SAA1 level is in blood, or optionally in serum, or optionally in plasma.

29. The oligomeric compound, composition or pharmaceutical composition of any one of claims 23-28, wherein a SAA1 nucleic acid comprises a pathogenic genetic variant, wherein the pathogenic genetic variant optionally is in an exon of a SAA1 nucleic acid, wherein the exon optionally is exon 3, wherein the pathogenic genetic variant optionally results in a Asp34Val amino acid substitution in a SAA1 protein, and wherein the pathogenic genetic variant optionally is c.101 A> T.

30. The oligomeric compound, composition or pharmaceutical composition of any one of claims 23-29, comprising: detecting presence or absence of a pathogenic genetic variant, and / or detecting presence, absence and / or amount of a SAA1 level, which optionally is an elevated SAA1 level.

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